An Epidemiological Study Of Nosocomial Infections At Mayo Hospital, Lahore
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Publisher: 2005 Dissertation note: The present study was designed to investigate the Prevalence of Etiological Agents of Nosocomial Infections in Mayo Hospital, Lahore-Pakistan of the 32,620 patients studied during 1997-2001; a total of 4502 (13.80%) patients acquired various types of nosocomial infections during their stay at Hospital. Clinical samples collected from various types of patients consisted of 1040 samples of Pus & Wound Swabs, 109 samples of blood; 115 of Pleural Fluids, 286 of Ascetic Fluids, 37 of Cerebrospinal Fluid, 1398 of Urine, 988 of Sputum; 329 of Burn Swabs, 99 of Patient Body Devices and 101 of Fecal and Drainage Material. The routine techniques for isolation. Identification through Biochemical, Serological and Antibiotic Sensitivity Testing were used for studying the Bacteriology of the selected samples. The present findings revealed that from a total of 4502 samples, 1287 Strains of Staphylococci, 429 Strains of Streptococci, 328 Strains of Enterococci, 781 Strains of Pseudomonas, 349 Strains of Enterobacter, 41 Strains of Acinetobacter, 26 Strains of Klebsiella, 140 Strains of Proteus, 1031 Strains of Escherichia, 67 Strains of Serratia, 93 Strains of Haemophilus, 119 Strains of other types of Gram Positive Bacteria, 13 Strains of other types of Gram Negative Bacteria, and 189 Strains of Yeast and Fungi were found as Etiological Agent for Nosocomial Infections.
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Pathogenesis Of Salmonellosis With Respect To Carrier States In Poultry And Its Public Health Impact
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; Literary form:
Publisher: 2006 Dissertation note: The present research endevour was made to study and investigate the prevalence of Salmonella enteritidis and Salmonella typhimurium from poultry feed, poultry meat and poultry eggs and their role in the chain of transmission of salmonellae to human beings. The objective was to generate data to improve the quality of poultry products and human health awareness.
Salmonellosis is one of the most wide spread food borne zoonoses. The etiological agents Salmonella enteritidis and Salmonella typhimurium not only' produce the disease but during the convalescent phase (after the recovery of disease) remain carriers for indefinite period of time. In this study 400 samples were collected and were distributed and detailed as; poultry feed (n=100), poultry intestines (n100 Small and n=100 Large intestines) and eggs (n=100) were collected for the identification of the organism through polymerase chain reaction (PCR). The Positivity percentage as tested through PCR for Salmonella enteritidis in the poultry feed was 20,15,10,15 and 10 for layer starter, layer grower, layer finisher, broiler starter and broiler finisher respectively (P>0.05). The positivity percentage as tested through PCR for Salmonella typhimurium for layer starter, layer grower, layer finisher, broiler starter and broiler finisher feed was 15,10,10, 10, and 10 respectively (P>0.05). There was no significant difference between layers feed and broilers feed as far as identification of salmonella enteritidis and salmonella typhimurium was concerned (P>0.05) but the prevalence range of salmonella enteritidis and salmonella typhimuilum from poultry feed was 10-20% which was biologically significant. The positivity percentage rate of Salmonella enteritidis for small and large intestine in Desi birds (local breed) was 2 and 16 % respectively. Where as for broilers in small and large intestine it was 4 and 18% respectively. The positivity of Salmonella typhimurium in small and large intestine of Desi birds was 2 and 14% where as in broilers it was 4 and 16% in the small and large intestine respectively. There was a significant difference (P <0.05) between the positivity of percentage of salmonella enteritidis and salmonella typhimurium as far as identification of Salmonellae from Desi and broiler meat was concerned.
It was found that 16%, 8%, 16'Y0 and 16% egg albumin was found positive for Salmonella enteritidis in layer egg albumin, Desi (local breed) eggj albumin, double yolk albumin and broken egg albumin respectively. In each case 25 egg albumin were collected and tested for the detection of Salmonellae. Similarly the egg yolk from layers, Desi (local breed) double yolk and broken eggs was taken and positivity rate for Salmonella enteritidis was found 12%, 4%, 12% and 12% respectively. It was found that 12%, 4%, 12% and 12% egg albumin was found positive for Salmonella lyphimurium in layer egg albumin, Desi egg albumin, double yolk albumin and broken egg albumin respectively. In each case 25 egg albumin were collected and tested for the' detection of Salmonella. Similarly the egg yolk from layers, desi double yolk and broken eggs was taken and positivity rate for Salmonella enteritidis was found 8%, 4%, 8% and 4% respectively. The positively rate for Salmonella typhimurium in both albumin and yolk was relatively less in both albumin and yolk of layers, desi double yolk and broken eggs. Statistically there was no significant difference (P> 0.05) but the prevalence of Salmonella enteritidis and Salmonella typhimurium from different eggs ranged between 4-16% and 4-12% respectively which was biologically significant.
The Salmonella enteritidis and Salmonella typhimurium were isolated, identified and grown on the artificial and selective media. The virulence of the organisms of Salmonella enteritidis and Salmonella typhimurium were estimated through calculation of LD50. It was found as 10358/mI and 103/ml for Salmonella enteritidis and Salmonella typhimurium respectively, having significant difference (P< 0.05). In order to understand the pathogenesis and carrier states of salmonella organisms in poultry, a group of 300 broiler birds were procured and divided into three groups were studied upto the age of 3 months. The infection was orally given on the 7th day of their age. As an average 86.74% of the birds were maintaining the organism of the Salmonella enteritidis in the large intestine during the entire experimental period in contrast to the small intestine in which 0% were found positive (P< 0.05). Similarly an average 94.94% of the birds were maintaining the organism of the Salmonella typhimurium in the large intestine during the entire experimental period in contrast to the small intestine in which 0% were found positive (P< 0.05) but non of the samples of Small and Large intestine of control group (Group-C) were found positive for Salmonella enleritidis and Salmonella typhimurium. There was a significant difference between Salmonella enteritidis and Salmonella typhimurium in large intestine of poultry (P< 0.05). The histopathology of different organs of broiler chickens i.e liver, lung, spleen, kidney, small intestine, large intestine, bursa of fabracious and lean muscles at different phases of disease was also conducted for the better understanding of pathogenesis due to salmonellosis. The principal lesions in the liver at the age of 14 to 28 days in groups A and B were leukocytic infiltration, necrosis and haemmorrhage. No lesions were recorded in liver after 28 days of age in groups A and B. No lesions were recorded in group C. The principal lesions of the lungs at the age of 14 to 28 days in groups A and B were leukocytic infiltration,' mild necrosis, vascular congestion and haemrnorrhages. No lesions were recorded in lungs after 28 days of age in groups A and B. No lesions were recorded in group C. The principal lesions of the spleen were mild leukocytic infiltration, necrosis and congestion at the age of 14 to 28 days in groups A and B. No lesions were recorded in spleen after 28 days of age in groups A and B. No lesions were found in group C. The principal lesions of the kidneys were marked tubutar necrosis with glomerular degeneration and Ieukocytic infiltration and haemmorrhages at the age of 14 to 28 in groups A and B. No lesions were1 recorded in kidneys after 28 days of age in groups A and B. No lesions were found in group C. The principal lesions of the small intestine were degeneration of mucosa with inflammatory cells, necrosis, inflammation, superficial ulceration on mucosal lining of intestine at the age of 14 to 21 days. No lesions were recorded in small intestine after 21 days of age in group A and B. No lesions were recorded in control group C. The principal lesions of the large intestine were leukocytic infiltration with necrosis and inflammation at the age of 14 to 91 days. The lesions were recorded up to 91 days of age in group A and B. No lesions were recorded in control group C. The principal lesions of Bursa of1, fabricious were atrophy & necrosis of bursal follicles and leukocytic infiltration at the age of 14 to 21 in groups A and B. No lesions were recorded in Bursa of fabricious after 21 days of age in groups A and B. No lesions were found in group C. The principal lesions of lean muscle were muscular degeneration and necrotic areas at the age of 14 to 21 days in groups A and B. No lesions were recorded in lean muscles after 21 days of age in groups A and B. No lesions were found in group C.
The carrier state was not only the source of spread of disease with in the poultry but also caused typhoid fever and food poisoning in humans. The chain of transmission started fron poultry feed to poultry meat and ultimately to humans as dead end host. Finally, the 400 samples of stool and blood from 200 human patients (100 suspected of typhoid fever and 100 suspected of food poisoning) were also collected from four different hospitals from urban area of Lahore for the identification of Salmonella enteritidis and Salmonella typhimurium through PCR method in order to see the public health impact of Salmonellosis through consuming the meat and eggs of the carrier birds. A total of 14% and 10% stool samples were found positive for Salmonella enteritidis and Salmonella Typhimurium in case of suspected typhoid fever patients respectively. Similarly 6% and 2% blood samples were found positive for Salmonella enteritidis and Salmonella Typhimurium. There was a significant difference (P< 0.05) in the sero positivity of stool and blood samples of suspected typhoid fever patients and also as for as Salmonella enteritidis and Salmonella typhimurium was concerned. However there was no significant difference (P> 0.05) between the hospitals On the average 14 and 10 stool samples were found positive against Salmonella enteritidis and Salmonella typhimurium from each of the 25 patients of each hospital respectively in case of suspected food poisoning patients. Similarly on an average 5% and 6% blood samples were found positive from 25 patients of each hospital respectively. There was a significant difference (P< 0.05) in the sero positivity of stool and blood samples of suspected food poisoning patients as far as Salmonella enteritidis and Salmonella typhimurium was concerned. However there was no significant difference (P> 0.05) between the hospitals.
A series of five experiments were conducted and carried out to study and explore the project Pathogenesis of Salmonellosis with respect to carrier states in poultry and its public health impact."
For this purpose, in the 1st phase, identification, isolation and characterization of Salmonella enteritidis and Salmonella typhimurium was attempted. It was followed by the estimation of LD 50 and carrier states and histopathological study at different phases of disease in broiler chickens experimentally infected with Salmonella enteritidis and Salmonella typhimurium to ascertain the nature of carrier states in terms of maintenance of the Salmonellae by different organs leading to histopathological changes and finally to the stage of shedding of the organism through the feces in the environment. Dissemination to human beings and the Public health impact of Salmonellosis was studied in the human subjects who consumed the meat and eggs of the carrier birds which were followed by testing their stool and blood samples through polymerase chain reaction (PCR). In this way the pathogenesis and chain of Salmonellas enteritidis and Salmonella typhimurium infection through poultry feed, meat, eggs and humans beings was transmissible. However, the humans were considered as dead end host. It was concluded that Salmonella enteritidis and Salmonella typhimurium was maintained in the large intestine of the poultry and has transmitted from poultry feed, poultry meat and poultry eggs to human beings and thus, causing typhoid fever and food poisoning.
Major aim of this research endeavour was to help in understanding the basic principles involved in the chain of infectious cycle of SalmoneUosis. In addition to that the application of the quality control of poultry products with respect to Salmonella infection to broiler chicks and broiler meat available in the market for human consumption is the ultimate goal of this project. The objective was to reduce the risk of Salmonellosis in poultry and humans. The following measures are suggested.
1. PREVENTION AND CONTROL OF SALMONELLOSIS IN POULTRY! ANIMALS
o The poultry and their environment should be monitored by frequente testing of Salmonellae.
o Bacteriological profile of poultry house environment.
o Serological testing of flock and removal of infected birds.
o Culturing of tissues from selected birds.
o Egg sheils, egg albumin & egg yolk culturing.
B. Hygiene and Sanitation
o Eggs from infected layer flocks should be pasteurized before consumption.
o Salmonella positive breeder flocks should be given pellet feed.
o Hatching sanitation
o Proper disinfection of hatching eggs.
o Proper sanitation and disinfection of farm premises.
o The provision of salmonella-free feed i.e pellet feed is of prime importance for the prevention of salmonella infections of poultry flocks and parent flocks.
o Control of rodent, insects and wild birds
o For routine treatment of eggs and progeny, only those antibiotics should be used that do not cause microbial resistance against drugs widely used in humans
o Resistance of Campylobacter spp, and Salmonella spp. to fluoroquinolones has become a public health risk. This does not exclude well targeted and transient use of antibiotics as essential measures in salmonellosis control programmes.
o Vaccination of breeder flock is recommended for decrease of the salmonella infection pressure.
1. MEASURES FOR THE PREVENTION AND CONTROL OF SALMONELLOSIS IN HUMANS
A. Meat and Eggs
o Wrap fresh meat in plastic bags at the market to prevent blood from1 dripping on other foods.
o Cook poultry products at temperature of 170°F for breast meat and at 180°F for thigh meat.
o Avoid eating raw or under cooked meat and egg.
o Cook poultry meat and egg thoroughly.
o Purchase only inspected grade AA eggs and animal food products.
o Handle raw eggs carefully:
o Keep eggs refrigerated
o Throw away cracked or dirty eggs.
o Do not eat half fried and half boiled eggs.
o Wash hands immediately after handling raw poultry or raw eggs.
o Full fried and full boiled eggs should be used for eating to prevent food borne Salmonellosis problem.
b. PERSONNEL HYGIENE MEASURES
o Washing of hands with soap and warm water before and after handling foods, after using the bath rooms.
o Refrigerate foods properly.
- Use bleach to wash cutting boards and counters used for preparation immediately after use to avoid cross contamination of other foods.
o People who have Salmonellosis should not prepare food for others.
o Educate the food handlers and persons who prepare food. Educational programmes covering pre- and post harvest food safety procedures, especially salmonella control, should be initiated in the animal and food production sectors for the public awareness.
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Isolation And Characterization Of Clostridium Perfringens From Domestic Animals An Man In Punjab
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Publisher: 2007 Dissertation note: The objectives of present investigation were to isolate the Cl. perfringens from the domestic and zoo animals and human beings; characterize it through biotyping and pathogencity observation, and to develop a vaccine- from the common CI. perfringens isolate. For this purpose a total of 1240 samples of morbid tissues (faecal samples from animals and gangrenous tissues from humans). From cattle (n=180), goats (n=180), horses (n=250), camel (n=250), deer (n=28), wild beast (n=07), monkeys (n16), zebra (n10), elephant (n01), yaks (n=09), foxes (n07), jackals (n=08), baboons (n=08), and bears (n08) were collected and processed for isolation of CI. perfringens. In addition a total of 100 human cases; 83 wound swabs and 17 gas gangrene were also collected and analyzed bacteriologically. This study has indicated that Clostridium (Cl.) perfringens causes multiple clinical problems in animals and human beings as was indicated by good rate of its isolation from the examined morbid tissues and fecal samples. Of the total 1240 samples from various types of animals 297 (23.95%) indicated the presence of CI. perfringens. The overall isolation percentages of various types of CI. perfringens from the cattle, sheep goat, horses, camel, wild beast, deer, bear, jackal, zebra, monkeys, yak, elephant, baboon, foxes, and humans were 22.2, 12.2, 57.2, 8.0, 21.6, 57.1, 30.76, 37.50, 50.0, 50.0, 37.50, 33.33, 100.00 75.00, 57.14 and 18.00, respectively. Of the tested population of domestic animals, goats indicated the highest Cl. perfringens (57.2%) infection rate. In the zoo animal population, the elephant, baboons, wild beast, jackals, and foxes were shown to be heavily infected with various CI. perfringens types compared to other wild life animals species. Of the 298 isolates obtained through this investigation Cl. perfringens type D was obtained from 118 (39.7%) morbid samples of the domestic and zoo animals; CI. perfringens type A from 63 (21 .21%) samples, Cl. perfringens type B from 95 (31.98%) samples; and the CI. perfringens type E was isolated from 21(7.07%) samples. None of the samples indicated the presence of CI. perfringens type C. Of the total 100 samples from the humans, CI. perfringens type A was isolated from 14 (14%) and Cl. perfringens type D was isolated from 04 (4%). None of the human samples showed the presence of Cl. perfringens types B, C, or E. Of the 17 human gangrene tissue samples, Cl. perfringens type A was isolated from 09 (52.94%) samples and the Cl. perfringens type D was recovered from 02 (11 .76%) samples. However, all attempts to isolate Cl. perfringens types B, C or E from the human gangrene tissue/material samples were unsuccessful.
The overall findings indicated that of the total 297 samples positive for various Cl. perfringens types 63 indicated the presence of Cl. perfringens type A. Of those 63 Cl. perfringens type A isolates, 49 were recovered from the animals; and 14 were isolated from the wound swabs and gangrene tissue material samples from humans. Of the 63 Cl. perfringens type A isolates from the animals, 5 were isolated from cattle; 3 from sheep, 20 from goats; 5 from the horses; 10 from camels, 01 from the deer; 01 from the zebra, 01 from baboon, 01 from fox, 01 from the monkey, and 01 isolate was recovered from yak. Of the 14 isolates of Cl. perfringens type A from humans, 05 were recovered from the open wound swabs, and 09 strains of the organism were isolated from the gangrenous tissue material.
Of the 297 samples positive for various Cl. Perfringens types, 95 animal samples indicated the presence of Cl. perfringens type B. These 95 isolates were obtained from cattle (n=22), sheep (n=10), goats (n=30), horses (n=03), camel (n=14), deer (n03), wild beast (n=02), monkey (n=02), zebra (n=02), yak (n=01), fox (n01), jackals (n02), baboon (n02) and bear (n=02). None of the human samples was positive for Cl. perfringens type B. Isolation of C/. perfringens type B from the zoo animals is a matter of concern for the human health, as the zoo visitors have the possibility to get infected with this organism.
Of the total 297 positive samples of faecal and morbid tissues from various types of animals and human being Cl. perfringens type D isolates were recovered from 118 (39.7%) samples. Of these 118 isolates of Cl. perfringens typeD, 114 were obtained from various types of animals, and 04 isolates were from the humans. Of the 114 animal isolates, 10 from the cattle, 5 from the sheep, 44 from the goats, 9 from the horses, 27 from the camel, 4 from the deer, 02 from the wild beast, 02 from the monkey, 02 from the zebra, 01 from the elephant, 01 from the yak, 02 from the fox, 02 from the jackals, 02 from the baboon, and 01 isolate the bear. A total of 04 CI. perfringens type D isolates were recovered from gangrenous tissue and open wound samples from human beings.
During this investigation 21 isolates of CI. perfringens Type E were obtained from domestic and zoo animals. Of the 21 isolates, 03 were from cattle, 04 from sheep, 09 from goats and 03 from horses, 01 from monkey, and 01 from the baboon. All the 21 isolations were from the fecal material of above mentioned animals. None of the human samples was positive for CI. perfringens type E.
Alpha toxin was produced by all of the 63 Cl. perfringens type A isolates. Within the toxin producing isolates, there was no difference in the quality of toxin in respect to its lethality for mice, dermonecrosis effects for guinea pigs and cytotoxicity in the HeLa cells. The 07 fecal isolates were hemolytic, lecithinase (+), and positive for all biochemical characteristics of Cl. perfringens. Those isolates were not lethal for mice, indicated no dermonecrotic activity in guinea pig, and produced mild degree of cytotoxicity in the cell cultures.
The activity of beta toxin obtained from 95 isolates of CI. perfringens type B isolates was determined using standard toxin-antitoxin test carried in mice and the standard serum neutralization test with antitoxin raised in rabbits. Within the toxin producing isolates, no difference was seen in the potential of toxin based on its lethality for mice.
Epsilon () toxin activity of the 114 isolates of CI. perfringens type D from animals and 4 of the human isolates was also determined. Of the 114 animal isolate, 110(96.49%), and all the 4 human isolates produced E-toxin. There was no difference in the lethal potential of toxin for mice, dermonecrotis action in guinea pig and production of CPE in VERO cells.
Iota (i) toxin activity of the 21 isolates of Cl. perfringens type E was also determined serum neutralization test in mice. Many isolates produced more than one major toxin. Ci. perfringens (CP) type
A produced Alpha (a) toxin; CP type B produced Alpha (a), Beta (3) and Epsilon (E) toxins; OP type D isolates produced Alpha (a) and Epsilon (E) toxins, and OP type E isolates produced Alpha (a) toxin + Iota (i) toxin.
The immunobiologic studies of isolates showed that many of the isolates were quite antigenic. Isolates of CI. perfringèns type D and B were found highly immunogenic as those isolates producing SN titer of 1:320.
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Identification And Genotyping Of Vp1 Genses Of Fmd Viruses
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Publisher: 2009 Dissertation note: Within two decades after its first report in 1954 from Pakistan, Foot and mouth disease has become endemic in the country and poses a serious threat to large as well as small ruminant population. Foot and Mouth Disease (FMD) is prevailing in cattle and buffaloes and is caused by either 0, A, Asia-i serotype of the FMD virus in Pakistan. The present study was undertaken to study the mutation rate of FMD virus and also molecular typing of the strains prevalent in Pakistan was done.
A total of 60 samples from buffalo and cattle were collected from five districts of Punjab including Lahore, Faisalabad, Sialkot, Okara and Sheikhupura. Soon after extraction of their RNA, all of them were reverse transcribed and then subjected to amplification by using different sets of the primers including universal as well as serotype specific primers. Then their VPI portions were amplified by using VP1 specific primers. Among 60 samples, 48 were positive with universal primers. Other 12 samples were not amplified with these primers hence not processed.
Among 48 FMD positive samples, 24 were positive with serotype 0 specific primers, 16 with serotype Asia-i and remaining 8 were positive with serotype A specific primers. After their amplification, the amplicons were run on the gel. These amplicons were extracted by using DNA extraction kit. After their purification, they were sent to Macrogen® (Seopl, Korea) and Centre of Excellence for Molecplar Biology, Pakistan (CEMB) for sequencing. Each amplicon was sequenced thrice and the consensus sequence was established eliminating sequencing errors.
Sequence identity and multiple sequence alignment of molecular sequences (nucleotide and amino acids) were performed with Clustal W algorithm (Thompson et al., 1994). Neighbour joining trees were constructed by using MEGA version 4.0 (Kumar et al., 2004). Nucleotide distance matrices were computed by Kimura two parameter algorithm based on the total nucleotide substitutions and evolutionary trees for VP1 genes were constructed.
For FMDV serotype '0' phylogenetic analysis, 14 VPI sequences from various field isolates were compared with some previously published Pakistani FMD 0 type VP1 specific sequences available with GeneBank and some recently published VP1 sequences reported by countries bordering with Pakistan including India, Iran and Afghanistan Similarly, 12 VP 1 sequences of FMDV serotype Asia-I isolates of this study were compared with previously published sequences and their phylogenetic relationship was established. However, the sequencing results of serotype A were inconclusive and were not included for phylogenetic analysis. Three sequences of three locally available FMD vaccines were also studied and compared with the outbreak strains.
Polymerase chain reaction was optimized with respect to MgCI2, buffer pH, annealing temperature, primer concentration, template concentration, and Taq polymerase. A concentration of 2.5 mM of MgCl2 resulted in the best amplification of the target sequences (Figure 1). The buffer with pH 8.8 yielded the best results (Figure 2) Although, the suggested annealing temperatures for various primers (of various serotypes) ranged from 48 °C to 63 °C, however, a temperature of 56 °C was found to be the best with all sets of primers (Figure 3). The best intensity DNA bands were observed with 0.3 pM concentration of the primers (Figure 4). Moreover, the best cDNA template concentration giving optimum amplification was found to be 3.0 p1 per reaction (Figure 5). Lastly, a concentration of 0.5 U of Taq polymerase was not sufficient for amplification of cDNAs, however, 1.0 U of enzyme was found to yield better amplification (Figure 6).
VP 1 DNA sequences of six previously published Pakistani FMD serotype 0 strains were analyzed phylogenetically with VP 1 DNA sequences of 14 isolates of the study. Serotype 0 isolates of this study distributed themselves into two distinct clusters (Figure 19). First cluster comprised of Sheikhupura 1 and 2, Muridkey 1, Raiwind 1, Nankana 1, Gujranwala 1 and Gujrat I isolates (Figures 19 and 20), whereas the second cluster included Depalpur 1, Sahiwal 1, Okara I, Multan 1, Toba 1, Faisalabad I and Pattoki 1 isolates (Figures 19 and 21). The first cluster was found to be associated with previously published Pakistani isolates of 2006 mostly. However, it also showed association with Afghanistan's isolates of 2004 (Figure 20). The second cluster seemed to be mostly related to previously published Pakistani isolates of 2003 (Figure 21). The overall grouping of the 14 sequences, when compared with each other, depicted a three clustered phylogram (Figure 22). Serotype 0 isolates from Depalpur, Sahiwal, Okara, Multan, Pattoki, Toba Tek Singh and Faisalabad grouped together into a clan and had more than 85% sequence similarity with each other. The second cluster consisted of isolates of Sheikhupura, Nankana, Raiwind and Muridkey. These sequences had more than 86% similarity with each other. The third cluster consisted of only two isolates which were 100 % similar to each other. However the third cluster had only 74 % sequence similarity to cluster I and 73 % sequence similarity when compared with cluster 2.
When the phylogenetic relationships with previously reported isolates of Asia 1 was evaluated, FMD Asia I isolates of this study were found to be scattered into two distinct groups (Figure 16). Group one consisted of isolates of Lodhran, Toba and Hafizabad that were more closely related to Indian isolates sharing more than 98% identity with each other and more than 94 % sequence identity with isolates of Indian 2001 to 2004 (Table 5 and Figures 16 and 17). However, they shared more than 86% sequence similarity with Pakistani isolates of 2002-2005 (Table 5). Group two comprised of isolates of kasur, Lahore, Pakpattan, Okara, Faisalabad, Jhang, Rahim Yar Khan, Bahawalpur and multan alongwith vaccine A and B (Figure 16). The isolates of group 2 were found to be closely associated with previously published isolates of Pakistani and Afghani origin of year 2003 and 2004 (Figures 16 and 18). Collectively, they shared an overall 70% sequence identity with each other. However, isolates of Bahawalpur, Rahim Yar Khan and Multan shared more than 98% similarity with each other, a measurement of close relationship denoting a likely common origin as one clan or dade. Similarly, isolates of Pakpatan, Faisalabad, Okara, Kasur, and Lahore shared 88% sequence identity with each other and qualified as one clade.
Although, overall amino acid sequence similarity of our isolates was not strikingly different from that of the published isolates, however, amino acid substitutions with dissimilar properties were found with a scattered pattern of distribution. For example, 15th amino acid residue which is hydrophilic in the previously published isolates had a substitution with a hydrophobic amino acid residue in our three isolates namely Sheikhupura 2, Muridkey I and Raiwind I (Figure 25). Similarly, 14th amino acid residue which is hydrophobic in nature was found to be replaced with a hydrophilic one in our last five isolates. Amino acid residue number 13 (Figure 25) had a substitution with a hydrophobic residue in some of our isolates etc. etc. It is interesting to note that such substitutions with amino acids having dissimilar properties have also been found, albeit at lower rate, in previously published sequences by many researchers (Figure 25).
A comparison of the deduced amino acid sequences in the critical VP I region of FMD serotype Asia I revealed that most of this study isolates shared very high homology with sequences of Vaccine A. However, the sequences of isolates of Lodhran, Hafizabad and Toba did not match much with that of either vaccines, A or B (Figure 23). Sequences of Vaccine A had a "K" which seemed to be replaced by a "T" in the sequences of most of the isolates. Considering the properties of various amino acids, this change does not signify a major shift in the three dimensional picture of the protein as K is a lysine, a positively charged amino acid, whereas a T is threonine, a hydrophilic amino acid in nature. Next substitution in most of the isolates is a "P" for "A" in comparison to the vaccines. Again, it is not a significant change as both P and A share the same property, hydorphobicity. Similarly a K with an R can be substituted without much change in the overall shape of the protein molecule. Next amino acid substitution is a leucine instead of methionine. Again both are hydrophobic in nature; hence their impact on the overall picture is minute, if at all. However, glycine and arginine are two very different amino acids; the former is a hydrophobic amino acid whereas the latter is positively charged one. Such amino acid substitutions may have the potential to make a major impact in terms of the epitopic differences in the capsids of vaccinal and field viruses. A comparison of the deduced amino acids of FMD serotype 0 isolates also exhibited such changes with the vaccinal virus (Figure 24).
Of the three hyper immune sera raised against three different vaccines in rabbits, only one vaccine induced a measureable immune response yielding good precipitation line against various FMD virus antigens.
In summary, RT-PCR for diagnosis of serotypes A, 0 and Asia 1 of FMDV was optimized and could be used for prompt and precise diagnosis of FMD in the country. Although, RT-PCR data pertains to bovines in the current project, but PCR optimization parameters are equally applicable to FMDV infections in other FMD susceptible animal species such as sheep and goat. The combination of PCR and sequencing of the VP1 gene to detect and analyze FMDV in disease outbreaks is fast (less than 6 hours for PCR and about 24 hours for sequencing), and it can give an accurate immunologic characterization of the virus, thus providing a rational basis for choice of vaccine. In fact, the molecular epidemiology of field isolates is a powerful tool to monitor the circulation of viruses (Saiz et al., 1993).
Secondly, various isolates of serotypes 0 and Asia 1 were sequenced along with some vaccinal strains. Sequence similarity tree analysis indicated that most of our isolates were closely related to previously reported Pakistani isolates and to those of neighboring countries such as India, Afghanistan and Iran. Additionally, amino acid sequence similarity data of major immunogenic site that forms 13G-13H loop in FMDV serotypes revealed that serotype Asia 1 vaccinal strain and Asia 1 isolates of this study possessed high degree of similarity suggesting a likely host immune response against the vaccine that may afford some protection against most field isolates of serotype Asia 1 type. Lastly, of three vaccines tested, only one was found to afford protection against field isolates of FMDV suggesting more work on vaccine issue in the country.
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Prevalence Of Caprine Mycoplasmosis In Different Areas Of Pakistan
Material type: Book ; Format:
Publisher: 2010 Dissertation note: A study was conducted to characterize caprine mycoplasma species and to know its prevalence in different areas of Pakistan during 2006 to 2007. For this purpose a total of 1440 different samples such as nasal discharge, pleural fluid, lung piece, synovial fluid, and milk samples (1180), and 260 serum samples were collected from clinically affected goats of different breeds, age and sex. These samples were collected from twelve districts including Mansehra, Peshwar, Swabi, Kohat, Abbottabad, Dera Ghazi Khan, Quetta, Pishin, Jhang, Sargodha, Lahore and Faisalabad with 6 union councils (UC) in each district. Twenty samples of different nature were collected from each of union council. These samples were subjected to cultural isolation, Growth inhibition test (GIT) using rabbit polyclonal antiserum against Mycoplasma mycoides subspecies capri, latex agglutination test (LAT) for the detection of Mycoplasma capricolum subspecies capripneumoniae and polymerase chain reaction (PCR). One twenty one samples out of 1180 showed turbidity in PPLO broth whereas out of these 121 samples 58 grew on PPLO agar. All 58 field isolated organisms showed positive reaction to GIT. None of the serum sample showed a positive reaction with LAT kit. Thirty five samples out of 1180 prior to culturing were positive for Mycoplasma mycoides cluster through PCR and identified as Mycoplasma mycoides subsp. capri (Mmc) through DNA sequencing, whereas 58 samples were positive with this technique after culturing. Prevalence of mycoplasmosis in hilly and plain areas (5.8 and 4.5 % respectively) is not significantly higher as compared to semi desert and sub hilly areas (3.3 and 2.9 % respectively) which may be due to chance alone. Furthermore, the adult group-3 (age > 1 year) has significantly lowest prevalence (2.7 %) of Mmc as compared to age group-1 (age < 181 days) with 5.1 % prevalance and age group-2 (age: 181 to 365 days) with 4.4 % prevalence. This difference may be due to chance but not areal difference. Similarly prevalence (4.7%) of mycoplasmosis in female goats is not significantly higher as compared to males (3.2%). Beetal, Piamiri, Beetal teddy cross, Baltistani and Desi breeds of goats showed higher prevalence only by chance as compared to other breeds in the areas under study. Saponin inactivated vaccine was prepared from this field strain and found to be effective against Mycoplasma mycoides subsp. capri in goats. This study focuses on characterizing the interaction of M. ovipneumoniae with ovine PBMC using carboxy-fluorescein-succinimidyl-ester (CFSE) loading and flow cytometry to measure lymphoid cell division. M. ovipneumoniae induced a strong in vitro polyclonal suppression of CD4+, CD8+, and B blood lymphocyte subsets. The suppressive activity could be destroyed by heating to 60 ºC, and partially impaired by formalin and binary ethyleneimine treatment that abolished its viability. The activity resided on the surface-exposed membrane protein fraction of the mycoplasma, since mild trypsin treatment not affecting viability was shown to reduce suppressive activity. Trypsintreated mycoplasma regained suppressive activity once the mycoplasma was allowed to re-synthesize its surface proteins. Implications for the design of vaccines against M. ovipneumoniae are discussed.
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Epidemiology Diagnosis And Chemotherpy Of Strangles In Equines
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Publisher: 2010 Dissertation note: Strangles is an infectious malady of equidae characterized by upper respiratory tract infection, dysponea, anorexia, regional suppurative lymphadenitis and causes high morbidity and low mortality. Considering the significance and utilization of equines in our country and the substantial losses rendered by Strangles, the present project was designed to study epidemiology, diagnosis and chemotherapy of strangles in Lahore and Sargodha districts of the Punjab province in Pakistan.
The present study comprised of five phases. In phase-I, epidemiology of the disease including prevalence, variations in SeM, SzPSe and Se18.9 proteins and mortality rate were studied in Lahore and Sargodha districts. For epidemiology, nasal swabs and pus samples from the affected lymph nodes of 500 equines (nr=250 horses, rutz250 mules) suspected for strangles were collected and cultured for identification of S. equl. The collected samples were processed at Medicine and Microbiology Laboratories of the University of Veterinary and Animal Sciences, Lhore, Pakistan and Gluck equine research center, Department of Veterinary Science, University of Kentucky, USA. Out of 250 horses and 250 mules, 113(45.2%) horses and 99 (3 9.6%) mules tested positive for S. equi. on the basis of culture. Number of S. equl isolates were significantly higher (P<0.05) in pus samples taken from sub-mandibular lymph nodes as compared to nasal discharge samples. The difference was significant (P<0.05) among mules of different age groups. The highest prevalence of strangles
was recorded in horses and mules less than 2 year of age as compared to those having age more than 2 years.
In the present study, prevalence of strangles round the year in horses and mules were also calculated and it was found to be the highest during the months of
February, March, April and May while few cases were seen during the months of January, June and July and no cases were observed during others months. The significant difference was observed (p<O.O5) among the prevalence levels of strangles in different months of the year. Similarly when compared the prevalence of strangles in different seasons of Pakistan i.e. summer, winter, spring and autumn. The highest prevalence rate was recorded during the spring season.
The prevalence on the basis of Polymerase chain reaction (PCR) of S. equi in horses and mules was also recorded. Out of 250 horses and 250 mules tested, 122(48.8%) horses and 113(45.2%) mules were positive for S. equi. When compared
the prevalence rate on the basis of PCR and culture of nasal and pus samples from affected submandibular lymph nodes it revealed that the sensitivity of Polymerase
chain reaction appears to be much greater than culture. The culture along with PCR is the best diagnostic technique for S. equi as PCR test does not differentiate between dead and live bacteria, hence a positive test may not correlate with active infection; therefore, a positive culture may be necessary to confirm the diagnosis.
In this phase of epidemiological study of disease, effect of selective pressure of allelic diversity in SeM of S. equi on immunoreactive proteins SzPSe and Se 18.9 was also studied. The aim of this study was to determine whether variations in SeM are accompanied by variations in the immunoreactive surface of exposed SzPSe and secreted Se18.9. Sequences of genes of 25 S. equi alleles isolated from different countries of the world over a period of 40 years were compared. Twenty different SeM alleles were identified including 6 not included in the data base (http:// pubmlst.org/szooepidemicus). Amino acid variation was also detected distal to the N- terminus of SeM. No variation was observed in SzPSe except for an Australian isolate which showed a deletion of one PEPK repeat. The Se 18.9 protein in all 25 isolates of S. equi did not exhibit any variation. Interestingly, only 2 SNP loci were detected in Se 18.9 compared to 93 and 49 in SeM and SzPSe respectively. The greater frequency of mutation in SzPSe compared to Se18.9 may be related to a high rate of recombination of SzPSe and the inclusion of exogenous DNA sequence based on the atypical GC percentage of its central hyper variable region.
In horses the mortality rate was recorded as 1.64% whereas the mortality rate in mules having less than 5 years of age was found to be 0.88%. No significant difference (P>0.05) in mortality rate among horses and mules of different age groups affected with strangles was observed.
In phase-I! of the present study, carrier status of the horses and mules were
studied. Out of 122 horses found positive to PCR, 20 horses (10<2 years and 10 between 2 and 5 years of age) were selected and monitored for 12 weeks. Their nasal
swab samples were used for identification of bacteria through culture and PCR on weekly basis. Till the end of 3rd week all horses <2 years of age remained positive but at the end of 4th to 7th weeks there remained positive only 5, 2, 1 and zero horses out of 10, respectively on the basis of culture whereas through PCR at the end of the 4th week all horse <2 years of age were found positive, but at the end of 5th to 10th weeks there remained 7, 5, 4, 2, 1 and zero horses out of 10, respectively. While all the horses aging between 2 to 5 year, were positive up to the 1St week but at the end of 2nd to 8th week out of 10 there were 9, 7, 6, 3, 1, 1 and zero horses respectively positive on the basis of culture but through PCR, all horses were positive till 4th week but at the end of 5th to 9th week number was reduced to 9, 7, 6, 3, 2 and zero. Similarly, out of 113 mules, 20 mules (10<2 year and 10 between 2 and 5 years of old) were also monitored for 12 weeks to study their carrier status. After the end of 2nd week all mules <2 years of age were positive but at the end of 3rd to 6th weeks there remained 7, 3, 1 and zero mules out of 10, respectively on the basis of culture but through PCR at the end of the 5th week all mules <2 years of age were positive, but at the end of 6th to 10th weeks there remained 9, 7, 3, 2 and zero mules out of 10, respectively. While in 2 and 5 year old mules, all were positive up to the 2nd week but at the end of 3rd to 7th weeks there were 6, 4, 2, 1, 1 and zero mules out of 10, respectively on the basis of culture but through PCR, all mules were positive up to 5th week but at the end of 6th to 10th weeks there were 8, 5, 2, 1 and zero. Horses and mules were declared free of infection on the basis of three consecutive negative samples through culture and PCR.
From the result of present study, it may be concluded that sensitivity of Polymerase Chain Reaction appears to be much greater than culture for study of carrier status of equines. Moreover, recovered animals should be kept in quarantine period at least upto 9th week because the recovered horses and mules remain carrier for prolonged period of time and can act as source of infection for susceptible animals through periodic shedding of S equi. (comprising 10 horses and 10 mules) for in-vivo trials. Efficacy of the antibiotics was assessed weekly on the basis of negative nasal swab culture. Results of in-vitro antibiotic sensitivity revealed that in horses and mules, S equi was most sensitive to Procaine penicillin followed by ceftiofur Na, cephradine, erythromycin, ampicillin, tetracycline, chloramphenicol, sulfamethoxazole, trimethoprim + sulfdiazine and gentamycin whereas the result of in-vivo antibiotic trials revealed that horses and mules suffered from strangles without abscess formation were most sensitive to Procaine penicillin followed by ceftiofur Na, cephradine and erythromycin whereas animals which developed abscess showed no response. It is concluded from the result of present study that Procaine penicillin is most effective in-vitro and in-vivo antibiotic followed by ceftiofur Na and cephradine. These antibiotics might be used for the treatment of strangles infection.
Phase-V, comprised over in-vitro trials of disinfectants. Efficacy of disinfectants, like povidone iodine, 0.6% H2S04, dettol and bleach was assessed. Phenol Co-efficient Test was applied, to ascertain efficacy of these disinfectants, used
in, in-vitro trials. Among four disinfectants, povidone iodine was found to be the best one with a phenol coefficient of 1.25 that is greater than phenol i.e. 1.00 while 0.6%
H2S04 showed similar phenol coefficient as that of phenol. The phenol coefficient of dettol and bleach were observed as 0.5 and 0.75 respectively. Therefore it is recommended that S. equi is highly sensitive to povidone iodine and 0.6% H2S04.
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Implications Of Varying Electrolytes (Sodium Potassium And Chloride And Their Balance On Growth Performance and Physiologcal Responses of Broilers
Material type: Book ; Format:
; Literary form:
Publisher: 2010 Dissertation note: A series of experiments were envisaged to evaluate the effect of supplementation of dietary electrolytes with applicability of dietary electrolyte balance by using different salts on growth and carcass responses, body physiological responses and litter condition of modern day broiler chickens under phase feeding system.
Day-old straight-run Hubbard broiler chicks were randomly allocated to eight dietary treatments replicated four times in such a way that a floor space of 0.09 m2 was provided to each bird. Birds were housed in environmental control system. Continuous light was provided 24 hours for the first 3 day and thereafter a light pattern of 23L:ID was adopted for the entire experimental. In each experiment, a basal diet was formulated having lowest level of each electrolyte. In experiment 1, Na and DEB in the basal diet were maintained at 0.08% and 160 mEq/kg, respectively. This basal diet was then supplemented with sodium bicarbonate (NaHCO3) and disodium sulphate (Na2SO4) to maintain four levels of Na (0.17, 0.26, 0.35, and 0.44%) by fixing K and Cl with DEB 200, 240, 280 and 320 mEq/kg, respectively. In experiment 2, a basal diet was prepared to contain the lowest level of K and DEB i.e. 0.70% and 160 mEq/kg, respectively. This basal diet was supplemented with potassium sulphate (K2S04) and potassium carbonate (K2C04) by fixing Na and Cl. So, four levels of K (0.86, 1.02, 1.18, and 1.34%) were maintained in eight dietary treatments. In experiment 3, a basal diet was prepared to contain the lowest level of Cl and DEI3 i.e. 0.17% and 320 rnEq/kg, respectively. This basal diet was supplemented with ammonium chloride (NH4CI) or calcium chloride (CaCl2), so that, in each diet, we can have the increase of 40 mEq/kg DEB at 0.3 I, 0.45, 0.59 and 0.73% of Cl at DEB 280, 240, 200 and 160 mEq/kg, respectively, by fixing Na and K.
At the end of each phase (pre-starter, starter, grower and finisher); data of feed intake, weight gain, feed to gain ratio, mortality, water intake, water intake-to-feed intake ratio and litter quality were collected and evaluated. At the end of each experiment, two birds were slaughtered for their carcass and body physiological responses. Blood was also collected from these same birds for blood pH. glucose and serum mineral analyses. For statistical analyses, four (4) levels of electrolyte were used with two (2) sources of salt in a factorial arrangement of 4 x 2 under completely randomized design using GLM.
In experiment 1, highest weight gain and feed intake were found in birds consuming 0.17% (NaHCO3) and 0.44% (Na2SO4) dNa, respectively during d 1-10. However during d 11-20, weight gain and feed:gain were reduced with same levels of dNa. Maximum weight gain was found in diets containing 0.17 and 0.24% dNa during d 21-33 and 34-42, respectively. Improved FG was the result of diets containing 0.20% (NaHCO3) and 0.37% (Na2SO4) dNa during d 2 1-33. Linear rise in water intake was observed in birds with increasing dNa during d 1-42. Minimum litter dampness was seen at 0.37% (NaHCO3) and 0.2 1% (Na2SO4) during d 1-10. Minimum and maximum mortality were observed at 0.37% level of dNa in case of supplementation of NaHCO3 and Na2SO4, respectively. Significantly increased pH and kidney weight while reduced dressing percentage were observed by amount and salt of dNa. Increased breast, thigh and gizzard weights were observed with increasing sodium. Weights of pancreas, gall bladder, bursa, and lungs, and shank length were affected by interaction of amount and salt of dNa.
In experiment 2, BWG (P0.03) and feed:gain (P0.05) was improved at 1.20% dK during 32 to 42 d of age. K2S04 supplemented diets increased feed intake during I to 10 d (P<0.05), water intake during 34 to 42 d (P0.04) and mortality during 1 to 42 d (PE0.02). Water intake was increased linearly with increasing dK when supplemented by K2C03 whereas this was decreased linearly with increasing dK with that of K2S04 during 11 to 20 d (P0.002). The K2S04 supplemented diets lowered the blood pH (P0.00l), dressing (P0.04), abdominal fat (P0.03) weights and shank length (P0.02). A significant salt x dK effect was observed where low levels of dK with K2C03 and high levels with K2504 exhibited lower litter moisture during all phases. Increasing concentration of serum cations was observed by increasing dK, by balancing of increasing serum HCO3 with decreasing Cl at the end of the experiment.
In experiment 3, body weight gain and water consumption were optimized at 0.73%, and 0.73% (CaCI2) and 0.45% (NH4CI), respectively, during d 1-10. During d 2 1-33, maximum weight gain and feed intake were observed at 0.42%, and 0.63% (CaCI2) and 0.63% (NH4CI), respectively. Highest weight gain (0.60% dcl), feed intake (0.61% CaCI2 0.42% NH4CI) and mortality (0.73%) while improved feed:gain (FG; 0.38% dCl) were obtained by interaction effects of amount and source of dCl during d 34-42. Fl (0.60%), feed:gain (0.3 8%) and litter moisture (0.31% NH4CI; 0.35 CaCl2) was affected during I -42d by amount of dcl. Increased blood pH, serum glucose and dressing percentage were found by dCl and replacing CaCI, with NH4C1. Improved breast meat, thigh meat and shank length while reduced abdominal fat were observed by replacing salts (CaCI2 withNH4Cl).
It is concluded that birds showed better growth performance and reduced mortality against high levels of dietary sodium in Na2SO4 than NaHCO3 supplemented diets, while significant rise in pH, breast and thigh meat yield while reduced dressing percentage were observed with increasing dietary sodium. The importance of high concentration of dK for better weight gain and feed efficiency was depicted in later stages of production. K2C03 increased survivability and dressing responses but both dK levels and salts played important role for water intake, litter condition, carcass characteristics and serum mineral concentration. Birds were also suggested to be more sensitive to amount and source of dC1 in later part of their life.
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Feeding Management For Optimum Growth, Reproduction And First Lactation Performance In Sahiwal Heifers
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Publisher: 2010 Dissertation note: Sahiwal is well known dairy cattle breed in the tropical and subtropical regions of world for its excellent heat and tick resistance. The value of adequate nutrition and management of replacement heifers is mostly overlooked and production losses linked with slow growth rate are not entirely realized. Efficient utilization of nutrients like energy during pre pubertal and gestation periods is needful for melioration. The study included two experiments. The aim in first experiment was to investigate the effect of varying dietary energy levels on pre pubertal growth and age at puberty in Sahiwal heifers. Twenty Sahiwal heifers (Age = 12 ± 2 month and avg. wt = 125 kg) were assigned to four dietary treatments having five animals on each treatment. Isonitrogenous (CP=13.7%) diets having varying energy levels, viz; A=100% (Control), B=88%, C=112% and D=124% of NRC recommended level for small breed non bred heifers were fed to the respective groups until onset of puberty.
Dry matter and protein intakes were not influenced by varying dietary energy levels during pre pubertal period. However, metabolizable energy (ME) 124% of NRC recommendation enhanced average daily gain (ADG) up to 571±15 g/d which was higher than all other dietary energy levels, whereas it was similar between ME 100% and ME 112% (442±11 and 450±05 g/d, respectively) but lower in ME 88% (397±07 g/d). The improvement in ADG of heifers fed ME 124% of NRC might be attributed to availability of excess energy nutrient for heifers to fulfill not only maintenance requirements but also to grow and develop body reserves. Provision of extra dietary energy improved efficiency of diets which might be attributed to availability of surplus dietary energy enabling heifers to convert feed into live body mass more efficiently. The 13 to 18 months of age was found optimum time period to have significantly highest ADG in Sahiwal heifers. This might be attributed to propitious physiological conditions under which heifers grow at faster rate.
The optimum increase in body structures (Body length, height and heart girth) was achieved in ME 124% of NRC recommendations. The phase from 13 to 18 months of age was found optimum possessing significantly highest values of increase in body length and heart girth, whereas phase from 19 months to age at puberty was optimum to achieve significantly highest body height. The optimum increase in heart girth during first two phases (13 to 19 months of age) might be attributed to relatively faster muscle growth in body than bone growth. The digestibility percentages of nutrients (DM, CP, NDF and ADF) were not influenced by different dietary energy levels. No influence of dietary energy levels on digestibility of nutrients in the present study might be attributed to best adaptability of Sahiwal heifers to utilize diets even with low energy under local environment.
Similarly, age at puberty was also not affected by dietary treatments and overall average was 833 ± 10 days. The optimum performance in terms of age at puberty at lower dietary energy level might be attributed to lesser energy requirements of Sahiwal under tropical and subtropical environment condition as elaborated by NRC (2000) that maintenance energy requirements of Bos indicus breeds including Sahiwal are about 10% lower. The similar pattern of influence was observed in serum progesterone concentration. The average of progesterone detected during a month before puberty was 0.44±0.005 ng/mL and during a month after onset of puberty was 1.48 ± 0.03 ng/mL serums. The similar rogesterone concentration among dietary treatments might be attributed to similar age at puberty in Sahiwal heifers.
It is concluded from results of first experiment that higher dietary energy level (ME 124% of NRC) enhanced growth parameters and feed efficiency but reproductive performance of Sahiwal heifers in terms of age at puberty was optimum even at lower dietary energy level (ME 88% of NRC recommended level) under local environment conditions of Pakistan.
The aim in second experiment was to study the effect of feeding varying dietary energy levels during last trimester of pregnancy on 1st lactation performance in Sahiwal heifers. Five to six months pregnant Sahiwal heifers (n=16) were assigned four dietary treatments having four heifers on each treatment. Iso-nitrogenous (CP=14.1%) diets having varying energy levels, viz; A=100% (Control), B=88%, C=112% and D=124% percent of NRC recommended level for pregnant heifers were fed to the respective groups until calving. After calving, all heifers were fed a similar diet having CP (16.2%) and ME (1.72 Mcal/kg).
Dry matter and CP intakes were similar across the dietary treatments. Pre calving ADG was not different among heifers fed ME 112 and ME 124% (486 ± 13 and 497 ± 05 g/d, respectively) but higher than other diets, whereas it was also higher (444 ± 07 g/d) in ME 100% than 397 ± 08 g/day in ME 88% of NRC recommendation. Feed efficiency was similar between ME 124 and ME 112% but higher than other diets, whereas ME 100% was also more efficient than ME 88% of NRC recommendation. The higher feed efficiency in higher dietary energy levels might be attributed to availability of surplus dietary energy enabling heifers to convert feed into live body mass more efficiently. Better body score through higher pre calving dietary energy level might be attributed to availability of energy for animal in surplus to its requirements of maintenance and pregnancy. Higher level of energy at this stage enabled pregnant heifers to develop extra body reserves needed in early lactation period to fulfill high demand of lactogenesis. The similar birth weight of newly born calves might be attributed to the factor that needs of conceptus (growth of fetus, fetal membranes, uterus and mammary glands) are accorded high priority by the homeorhetic controls it transmits to the dam.
Extra energy levels beyond NRC recommendation during prepartum period were not advantageous to increase milk yield in 1st calf heifers. The performance of 1st calf heifers in terms of milk yield was only optimum through pre calving feeding according to NRC recommendations. The lesser milk yield in diets having higher energy levels than recommended by NRC might be attributed to more availability of mammary fat pad which may limit further parenchymal tissue development and consequently decrease milk yield during subsequent lactation. However, milk fat percentage increased as pre calving dietary energy level was increased, whereas milk protein, lactose and SNF percent among animals fed different experimental diets did not differ. It is concluded from results of second experiment that the optimal performance of pregnant Sahiwal heifers was achieved through provision of pre calving extra dietary energy (ME 112%) beyond the NRC recommendation but first lactation yield was found optimum in heifers fed diet having energy level as per recommendations of NRC.
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Epidemiology And Controls Of Coccidiosis In Cattle
Material type: Book ; Format:
Publisher: 2009 Dissertation note: Field study was conducted from September, 2007 to August, 2008 and a total of 2700 rectal faecal samples were collected from cattle farms of 3 categories i.e. Government Dairy Farm, Military Dairy Farm and Peri Urban Dairy Farms (Gawala Colonies) Lahore. Seventy five random samples were collected from each category of farms on monthly basis. The results of field study showed that overall prevalence of coccidiosis in cattle was 54.55%. Prevalence of coccidiosis in cattle at Military Dairy Farm Lahore was the highest (65.33%) during Autumn followed by summer (52.66%) then winter (47.66%) whereas the lowest (34.00%) during spring season. The highest (56.66%) prevalence was observed in animals between 6 & 12 month, whereas the lowest (46.33%) in animals under 6 months age. Prevalence of coccidiosis above 1 year of age was 50.66%. No coccidial oocysts was detected in calves less than 15 days old. In female animals prevalence was 51.22%.In the present study, the maximum oocyst per gram of feces (OPG) count was 65,000 whereas the minimum count was as 2000. The count was variable in different age groups and found to be decreasing in adult animals. The mean OPG in group A (under 6 month), group B (6 month to one year) and C (above one year) was 44000, 38000, and 22000, respectively. The four species of Eimeria were identified in all age groups i.e. E.bovis (29.28%) E.zuernii (26.03%) E. cylindrica (23.42%), E. ellipsoidalis (21.25%).
The results of field study showed that prevalence of coccidiosis at Government Dairy Farm, Lahore was the highest during autumn (49.33%), followed by summer (44.33%), then winter (38.33%) where as the lowest during spring (30.33%). The highest (62.66%) month wise prevalence of coccidiosis was noted during August whereas the lowest (28.00%) during April. The highest ( 45.33%) prevalence of coccidiosis was observed in animals aged between 6 to 12 months, followed by 41.35% in animals under 6 months of age whereas the lowest (36.00%) above I year. Female animals were more frequently affected (41.28%) than males (39.50%).In the present study, the maximum OPG count observed was 55,000 and the minimum count as 2500. The counts were variable in different age groups and found to be decreasing in adult animals. The mean OPG of group A, B, C was 42,000, 35,000 and 20,000 . In the present study five species of Eimeria were E.bovis. E. zuernii E. cylindrica, E. subspherica, E. ellipsoidalis.
The results of field study showed that prevalence of coccidiosis at Peri Urban Dairy Farms (Gawala colonies), Lahore was 71.55%. Month wise prevalence was the highest during August (90.66%) whereas the lowest (48%) during April. The seasonal prevalence indicated that it was the highest during autumn (84.00%), followed by summer (78.33 %), then winter (69.33%) whereas the lowest during spring (50.00%). The highest prevalence of coccidiosis (80.66%) was observed in animals under 6 months of age, whereas the lowest (62.33%) in animals above I year. Prevalence of coccidiosis in animals aged between 6months to 1 year was 71.66%. No coccidial oocysts were detected in calves less than 25 days old. Prevalence of coccidiosis was higher (74.61%) in females than in males (63.60%). In this study, the maximum OPG count observed was 65,000 and the minimum count as 2800. The counts were variable in different age groups and found to be decreasing in adult animals. The mean OPG of group A, B,C was 48,000, 38,000 and 23,000 respectively.
Age wise analysis of Eimeria species showed that above mentioned five species were found in all age groups and most predominant species was E.bovis (26.39%) followed by E. zuernii (19.87%), E. cylindrica (23.60%), E.ellipsoidalis (18.63%), whereas the lowest prevalence of E.subspherica (11.49%)was noted (Table 16). The counts were variable in different age groups and found to be decreasing in adult animals. There was inverse correlation of OPG and the age of animals.
The overall prevalence of coccidiosis was the highest during autumn (66.22%) followed by summer (59.66 %) then winter (51.77%) whereas the lowest in spring (38.22). The role of Meteorological data i.e. temperature, humidity and rain fall on the prevalence of disease was also studied. The bionomical showed that humidity and rain fall played a very important role in the causation and spread of disease and also help in the development of sporulated oocyst. Increased temperature showed higher prevalence of disease.
The results of histopatholgical studies showed that there was an increase cellular infiltration of leukocytes, cellular debris in most of intestinal portion.
Results of therapeutic trials by using toltrazuril, amprolium, sulphaquinoxaline, lasalocid are presented in table 17. The result of therapeutic trials showed that efficacy of toltrazuril was better than amprolium, sulphaquinoxaline and lasalocid. No clinical signs of disease were observed in treated animals while in diseased animals signs of disease were observed i.e. animals showed diarrhoea, loss of weight gain. From the results it was noted that efficacy of toltrazuril was better than other drugs . Statistically, there was no significant difference between efficacies of all four drugs. The efficacy of per oxygen based disinfectant was higher as compare to oocide while non- treated animals showed clinical signs of disease. Statistically, there was no significant difference between efficacies of both disinfectants
Result of chemo prophylactic products are presented in table 19. It was noted that sonicated vaccine showed high antibody titer as compare to non- sonicated vaccine.
Result of the challenge experiments revealed that the inactivated sonicated vaccines gave 100% protection to the challenge calves. Their faeces were normal and no clinical sign was recorded even 42 days post vaccination. Few remaining live oocysts were not able to produce the disease in calves.
The weight gain of treated animals was higher as compare to non-treated animals. The FCR value in treated animals was better than non treated animals
" Prevalence of coccidiosis was the highest during autumn followed by summer where as the lowest during spring. Farm wise prevalence of coccidiosis indicated that it was higher in Peri Urban Dairy Farms followed by Military Dairy farm where as the lowest at Government Dairy farm.
" Prevalence of coccidiosis was higher in calves below 9 months of age than above 9 months. All the animals examined for coccidian were naturally infected with coccidiosis. These animals were not experimental calves and prevelance of infection was based on random selection of animals. Overall Prevalence of coccidiosis was slightly higher in females than male. Species wise prevalence indicated that Eimeria bovis is more pathogenic than other species.
" Results of chemotherapeutic trials showed that among the four drugs used i.e. Toltrazuril, Amprolium, Sulfaquinoxaline and Lasalocid. Toltrazuril showed the highest efficacy followed by Amprolium, where as Lasalocid showed the lowest efficacy. No side effects of these drugs were noted when were given at their recommended dose rate and marked clinical improvement in animals was noted after treatment.
" Two disinfectants were tried. Per oxygen based disinfectant showed better results than. Oocide disinfectant.
" Histopathological studies showed inflammatory granulocytic infiltration of the mucosa and cellular debris in most of intestinal portions. There were necrosis of villi and degeneration of villi. Haemorrhages in mucosa and sub-mucosa were noted. Some of the glands in the sub-mucosa of intestine showed degeneration & necrosis.
" Indirect Haemagglutination (IHA) antibody titer was higher in calves vaccinated with inactivated sonicated vaccines as compared to the calves vaccinated with inactivated sporulated vaccine. Results of the challenge experiments revealed that the inactivated sonicated vaccines gave protection to the challenge calves. Their faeces were normal and no clinical sign of disease were observed even 42 days post vaccination.
" Weight in infected group was reduced. After treatment, high weight gain was reported in treated animals than control group.
" Overcrowding should be avoided.
" Provide good hygienic and managemental conditions in farms.
" Proper drainage of rain.
" Feeders and wateres should be above the level of the ground.
" Regular use of coccidiostats is the need of the day.
" Diseased animals particularly with diarrhoea should be separated from healthy animals.
" Stocking density should be according to recommended of world Association of Parsitologists.
" Contaminated faeces should be properly disposed off.
" Grazing of animals during rainy season should be avoided.
" Animals should be provided well balanced nutritive food.
" Entry of visitors in the livestock farms should be restricted
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Comparative Growth Rate And Body Composittion Of Major Carps (Labio Rohita , Cata Catla And Cirhinus Mrigala )
Material type: Book ; Format:
; Literary form:
Publisher: 2011 Dissertation note: Presently fish culture in Pakistan is primarily dependent on natural food produced in pond by the application of organic and inorganic fertilizers. It is supplemented with cheaper agriculture by-products to meet the nutrient deficiencies. Artificial feed which is a blend of various plant and animal by-products is rarely used. Development .of appropriate artificial feed now has become mandatory to transform conventional fish culture practices to advanced fish production systems to improve per unit fish production. The present study was therefore signed to formulate a quality supplementary feed from cheap and easily available feed ingredients that contains at least minimum required nutrients for different age groups (fingerlings and grow-out). The feeds developed during these studies were evaluated in terms of growth, diet utilizalion efficiency and its effect on the body composition and flesh quality of the three Indian majr carps (Catla cat/a, Labeo rohita and Cirrhinus mrigala).The study comprised of three trials. Trial I was conducted on fingerlings of individual species under monoculture system using 42% protein diet. Trials II and III were conducted on Grow-out fish using 35% protein diet under monoculture and polyculture systems. The study was conducted in earthen ponds having an area of 0.03 ha with three replicates and a control. After preliminary preparation of ponds, in trial I, fingerlings were stocked at 80 fish per pond. while in trial II at 70 fish of each species and in trial III ratio of 30%, 50% and 20% of Catla catla, Labeo rohita and Cirrhinus mrigala per pond were maintained. All the ponds received same amount of organic and inorganic fertilizers (cow dung, poultry manure, SSP and urea) thoughout the experimental period. Supplementary feed in trial I was applied at 4% of fish wet body while in trial II and III feed was applied at 3% of fish wet body weight daily. In trial I 42% protein diet was used containing fish meal. soybean meal. maize gluten (60%). rice polish, wheat bran. maize grains. molasses. vitamins and minerals while in trial II and III 3YYo protein diet containing fish meal, soybean meal. canola meal. rice polish. wheat bran, molasses, vitamins and mineral was used. Growth parameters in terms of length and weight gam were regularly monitored fortnightly. Organolept sensory evaluation was done at the termination of each trial. Proximate fish body composition was determined at the start and at the end of the experimental trials. Fatty acid profile of three experiments was performed at the post-trial basis. In addition, specific growth rate (SGR), feed conversion ratio (FeR). protein efficiency ratio (PER). protein utilization (PU). gross nitrogen retention efficiency (G RE %) and gross energy retention efficiency (GERE %) were also determined. Proximate analysis of feed ingredients and formulated diets was also done. Key physico-chemical parameters viz. temperature, dissolved oxygen (DO), free CO2, pH, electrical conductivity, total dissolved solids, light penetration. salinity and nitrates, were regularly monitored during the study period. In trial I the highest net weight gain was observed in treatment group (D 1) (Catla calla 9425.83 g and 171.5 mm) followed by Labeo rohita (374.34 g and 178.7 mm) and Cirrhinus mrigala (288.18 g and 161.9mm). The lowest growth was observed in Cirrhinus mrigala (176.9 g and 116.4 mm) in control (DO). A significant difference was observed regarding net weight gain among three fish species and between different treatments (DO and 0 I). The net weight gain was significantly higher in trial I treated (01) ponds than control (~O). Percent weight gain and specific growth rate (SGR %) were also determined. Labeo rohita exhibited higher values (1762.51 % and 3.03%) followed by Catla calla (1341.58% and 2.95%), while Cirrhinus mrigala showed lowest (976.17% and 2.6%) with experimental diet (DI) Again Cirrhinus mrigala exhibited lowest percent weight gain and SGR (300.85% and '1.54%)in control (DO) ponds. In trial II grow-out under monoculture the net weight gain of fish differed significantly among three fish species and between treatments (DO and D2). Calla catla showed highest net weight gain (37\.88 g and 72.2 mm) followed by Labeo rohita (310.18 g and 72.3 mm) and Cirrhinus mrigala (270.75 g and 57 mm) in experimental unit (02) while a lowest net weight gain of Cirrhinus mrigala (162.15 g and 36.5 mrn ) was observed in control (DO). Percent weight gain and specific growth rate of three fish species Catla catla, Cirrhinus mrigala and Labeo rohita under different treatments were found non-significant. Although Catla catla showed highest percent weight gain and SGR values (109.78% and 0.81 %) followed by Labeo rohita (90.93% and 0.69%) and Cirrhinus mrigala (84.3% and 0.65%), respectively with experimental diet (D2). Lowest values of percent weight gain and SGR (48.54% and 0.43%) were observed for Cirrhinus mrigala in control ponds (DO). In trial III grow-out under poly culture the average final weight of fish was significantly different in control (~O) and experimental diets (02) while species showed non-significant difference regarding final weight and net weight gain. The highest final and net weight gain of Lobeo rohita (679.46 g and 370.5 g) followed by Cirrhinus mrigala (674.52 g and 303.86 g ) and Catla catla (607.2 and 307.06 g), respectively in experimental unit (D2) while Catla catla exhibited lowest final weight and net gain in weight (493 g and 182.3 g) in control (DO). Regarding percent weight gain and specific growth rate of three fish species under polyculture system no significant difference was observed hence, Labeo rohita showed highest percent weight gain and SGR (126.87% and 0.9%) followed by Catla catla (l 02.31 % and 0.76%) and Cirrhinus mrigala (85.15% and 0.63%), respectively with experimental diet, while Cirrhinus mrigala once again showed lowest values (40.12% and 0.37%), respectively in control diet (DO). Feed conversion ratio (FCR), protein efficiency ratio (PER), protein utilization (PU), gross nitrogen retention efficiency (GNRE %) and gross energy retention efficiency (GERE %), in all the three experiments under monoculture as well as in polyculture system, for fingerlings and grow-out fish of three species were found non-significantly different. However, in trial I fingerlings better FCR values (1.63, 1.56 and 1.43) were obtained for Catla catla, Cirrhinus Mrigala and Labeo rohita. Regarding gross nitrogen retention efficiency Catla catla showed highest GNRE % value (10.4) followed by Labeo rohita (9.3) and were found significantly different from Cirrhinus mrigala (6.5) in experimental unit. In trial II grow-out monoculture, FCR values 3.7. 4.57 and 4.56 for Calla calla. Cirrhinus mrigala and Labeo rohita were pbtained while GNRE % varied 9.5,5.8 and 8.0. respectively. In trial III grow-out poIyculture the FCR values of three species varied from 3.99, 4.72 and 3.61, respectively while GNRE % varied from 10.3, 8.2 and 12.5%, respectively among Calla catla, Cirrhinus mrigala and Labeo Rohita. The Labeo rohita for GNRE% differed significantly from other two species. No significant difference among species and between diets (DO, D 1 and D2) was observed in proximate composition in all the three experiments. However, in case of fingerlings Labeo rohita under experimental diet (D 1) showed higher protein contents (16.44<Yo) while Catla catla showed the lowest protein content (12.9%). Crude fat contents were found highest (7.28 %) in Labeo rohita with control diet (DO) followed by Cirrhinus mrigala (6.96 %) and Labeo rohita (6.S2 %) in experimental diet (01) while lowest values were observed for Calla catla (4.17%) in control (DO). The Ash contents showed minor variations among species and treatments ranged from (4.81 % and 3.S6%) for Catla catla, (4.34% and 4.7S%) for Cirrhinus mrigala and (3.98% and 4.49%) for Labeo rohita in control and treated ponds, respectively. Highest gross energy was found (6.S3MJg'l) for Labeo rohita and lowest (S.OMJg'l) for Catla catla with experimental diet (D 1). In trial II grow-out monoculture the highest crude protein contents (1S .16%) were observed in Labeo rohita followed by Cirrhinus mrigala (14.S3%) with control diet (~O) while lowest for Labeo rohita (12.13%) in (02). Higher contents of crude fat (7.31 %) were observed in Cirrhinus mrigala followed by Catla catla (S.38%) in experimental group and lowest amount 3.18% and 3.19% was observed for Cirrhinus mrigala and Catla catla in control group (~O) . . Higher amount 4.11 % was found in Catla catla under control (~O) while lowest amount 3.1 % was observed in Labeo rohita under experimental diet (D2). Highest gross energy percentage 996.13%) was observed for Cirrhinus mrigala under experimental diet (D2) while lowest 4.91 % was observed for Catla catla in control group (DO). In case of experiment III grow-out polyculture the proximate body composition highest crude protein contents (IS.76% and 10.53%) were observed for Cirrhinus mrigala followed by catla catla 911.87% and 13.3S%) and Labeo rohita (12.72% and 6.S6%) in treated (D2) and control (DO) group. respectively. Higher crude fat contents (6.S7%) were observed in Cirrhinus mrigala under (D2) while lowest (3.13%) in Labeo rohita and (2.9S%) in Catla catla. Ash percentage was found higher in Catla catla and lowest (2.14%) in Labeo rohita and Cirrhinus mrigala (2.87%) under (DO). Gross energy contents were found highest (6.84MJg,l) in catlacalla under (DO) and (6.56MJg,l) Cirrhinus mrigala under (D2) while lowest amount (3.24MJg.l) were observed in Labeo rohita under (DO). Mineral composition of three fish species under three dfferent experiments showed non- sign ificant differences. Minor variation regarding mineral composition was observed in pre- treatment and post-treatment level. However. Ca and P contents showed relatively higher percentage than Mg and K contents in all the three experiments. A significant difference was observed in Mg contents in experiment III where Catla catla showed significantly higher (0.045%) percentage than Cirrhinus mrigala and Labeo rohita each containing 0.02%. A significant difference was observed in fatty acid profile among three fish species and between diets (~O, Oland D2). Among fatty acids, palmitic acid (C 16:0) was found a dominating fatty acids in all the three experiments. In trial I highest concentration (40.59 g 100 g-1 was found in Cirrhinus mrigala under (DO) and 37.19 in (D1) while lowest (30.75 and 30.78 g 100 g.l) in Labeo rohita and Catla catla under (D 1). The concentration of total saturated fatty acids were observed higher and ranged from (40.20 to 53.29 g 100 g-I) followed by total monounsaturated fatty acids (29.30 to 37.81 g 100 g-I), w-6 PUFA (7.65 to 14.94 g 100 g') and @-3 PUFA (7.76 to 11.07 g 100 g-I). respectively. In case of trial II significant differences were also found among three fish species and diets (D0 and 02) for different fatty acids composition. Palmitic acid (C 16:0) also showed highest concentration ranged from 28.36 to 29.73 g 100 g-I). Total saturated fatty acids were found higher that varied from (35.90 to 39.41 g 100 g-I) followed by total monounsaturated fatty acids (36.52 to 40.84 g 100 g-I), and l:PUFA (19.02 to 24.40 g 100 g-I), respectively. In trial III once again same pattern of dominance of palmitic acid along with total saturated fatty acids (36.43 to 42.24 g 100 g-I) followed by total monounsaturated fatty acids (36.899 to 43.72 g 100 g-I) and 2:PUFA (14.97 to 23.03 g 100 g-I) were observed. In case of organoleptic evaluation all the species under di Iferent culture system and treatments illustrated non-significant differences. Hence. significant differences were observed among different cooking processes (steamed and fried fish). The physico-chemical parameters of pond water remained within the acceptable limit for Fish gowth. Although comparatively lower values of temperature were found for experiment II and III for grow-out trial that was conducted in fall. The correlation co-efficient studies revealed a positive significant correlation of temperature, TDS, light penetration and salinity with growth of fish species while pH showed positive non-significant correlation with growth of fish. It was concluded from the present study that both the experimental diets D I and 02 for different age groups (fingerlings and grow-out) showed significantly higher growth of all the three species in monoculture system. The diet D2 did not showed any significant higher growth in polyculture system but overall growth performance remained high in polyculture than monoculture treated ponds of grow-out fish. Comparison of species indicated that artificial diets (DI and D2) remained much suitable for Catla catla and Labeo rohita than Cirrhinus mrigala under both the culture systems. Non-significant difference was observed in the body composition and flesh quality irrespective of their economic viability. Information derived from the present research experiments will be useful in future research and formulating supplementary feed for Indian major craps for different age groups. It can also be helpful in understanding the mineral and fatty aeid profiles of the Indian major carps cultured under semi-intensive pond culure system whieh is first study of its kind on these species in Pakistan.
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Surveillance Of Tuberculosis In Buffaloes, Cattle And Derectton Of Mycobacterium Bovis And Mycobacterium Tuberculosis in Food of Animal Origin
Material type: Book ; Format:
; Literary form:
Publisher: 2010 Dissertation note: The main objectives of this study were: to survey the prevalence of TB infection in livestock and their products in Pakistan; to standardize PCR based techniques for the detection of TB in buffaloes, cattle and animal products (milk and meat) as presently no such system has been developed for the detection of TB in animals and their products in Pakistan; to evaluate improved tests for the differentiation of Mycobacterium complex isolates in cattle, buffaloes and animal food products and to compare modern and conventional methods for rapid diagnosis of the Mycobacterial spp. The study was performed in different experiments to have surveillance of tuberculosis in Buffaloes and Cattle; and to detect the presence of different Mycobacteria in animal food products. One thousand animals from different areas of Lahore District were screened with the tuberculin test. The milk and blood of tuberculin tested animals were further studied for the presence of Mycobacterial spp. by conventional methods as well as Polymerase Chain Reaction (PCR). In other experiments one hundred market milk samples and ten thousand five hundred tissue samples from twenty-one hundred carcasses at Lahore slaughter house were screened with conventional microbiological tests and multiplex PCR for differentiation of Mycobacterium species. The results indicated that PCR had more sensitivity and required less time to detect and differentiate different Mycobacterial species as compared to conventional methods. It was also noted that M. bovis were found in milk and blood of milking animals as well as tissue sample collected from Lahore slaughter house. On the basis of findings, regular monitoring of the milking animals, animals to be slaughtered, and workers handling these animals is suggested. It is also recommended to review the current slaughter act to prevent the slaughtering of TB affected animals.
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Studies On Cyanide Toxicity In Ruminants
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Publisher: 2011 Dissertation note: The present study was conducted with the objectives to: (a) determine the cyanogenic potential of various livestock fodder and grasses, (b) determine CN? content in blood of ruminants feeding cyanogenic plants, (c) develop a simple, reliable and inexpensive assay for the determination of CN? in blood, (d) evaluate the efficacy of various antidotes against CN? toxicity in a rabbit model, (e) find out the effect of CN? on hematological and biochemical profile, (f) study the postmortem and histopathological changes associated with CN? toxicity in various organs and (g) study alterations in tissue oxygenation and metabolic variables during acute CN? toxicity in pigs. For this purpose a total of 500 samples of various plants being used as fodder to livestock were collected from the field and analyzed for CN? content spectrophotometerically. To develop a simple picrate method for CN? estimation in blood, two goats were infused with KCN at 0.6mg/kg for 1 hours. Blood samples were collected at intervals and were subjected to the general method. Standard curve was developed using standard solutions of various concentrations of CN?. Additionally 6 rabbits were also given CN? orally for 40 days and blood samples were collected for CN? estimation. Later on, this method was successfully used to determine CN? levels from 500 blood samples of cattle, buffaloes, sheep and goats (n=125 each) feeding cyanogenic plants which were collected from the field. For antidotal studies forty two rabbits were randomly divided into seven groups viz. A, B, C, D, E, F and G each comprising of six animals. Rabbits in group A were given feed only and served as negative control, while the rabbits in group B received feed plus oral solution of potassium cyanide (KCN) and were positive control. Animals in group C were given feed, KCN and intraperitoneal (IP) injection of garlic extract. Rabbits in group D were treated with feed, KCN and IP injection of sodium thiosulfate (STS). Members in group E received feed, KCN and IP injection of both garlic extract and sodium nitrite (SNT). Animals in group F were treated with feed, KCN and IP injection of both STS and SNT whereas the rabbits in group G were given feed, KCN and hydroxocobalamin IP. The treatments were given to respective groups for a period of 40 days. At the end of 40 days, serum and fresh urine samples were drawn from each rabbit to study biochemical panel. Subsequently the rabbits were euthanized for postmortem and histopathological changes in various organs. For hematological and growth rate studies 12 rabbits were divided into two groups of six viz. A and B. Rabbits in group A were given feed only while members in group B were treated with feed and oral KCN at 3mg/kg for 40 days. The animals were weighed after every 10 days. Feed consumption rate, feed efficiency and weight gain for the members of each group were recorded. Blood samples were also collected for hematological studies at 0, 10, 20, 30 and 40 days. To study tissue oxygen and metabolic variables during acute CN? toxicity, 26 piglets were anesthetized. The non-invasive monitors were used to measured oxygen saturation, heart rate and rhythm and cerebral response to sedation. The invasive monitors were placed to measured beat to beat variability of the arterial blood pressure, pulmonary artery pressures. Mixed venous oxygen saturation (SvO2) and cardiac output were continuously monitored. Regional brain O2 (cerebral cortex) and skeletal muscle O2 saturation (rSO2) were also measured via sensors. After getting baseline metabolic and hemodynamic measurements that included arterial and venous blood gas analysis, lactates and cyanide levels, all pigs were started on an infusion of NaCN (0.55 mg/kg/hr). The infusion continued until the occurrence of sustained apnea (?3 minutes). A non-significant difference was found in the CN? content of Sorghum bicolor and S. sudanese, while a significant difference was observed between the CN? content of Jumbo grass (S. bicolor x S. Sudanese hybrid) and S. halepense. Jumbo grass and S. halepense were found to have significantly higher CN? concentrations than S. bicolor or S. Sudanese. Maize has significantly lower CN? content compared to S. bicolor, S. sudanese, Jumbo grass, or S. halepense. A gradual increase in CN? content with increasing height of plants was also observed. Highest CN? levels were observed at heights of 91-100 cm in sorghum varieties. At greater heights, a gradual decrease in CN? content was seen in all plant species, with the lowest levels at heights of 200 cm or above. No CN? was detected in maize at heights over 131 cm. In picrate method, the calibration curve was linear (R2=0.99) in the range of 0.3-120 mg CN?/L. In standard CN? solutions a color change in picrate paper from yellow to brown was observed at a concentration of 3 mg CN?/L and above. This method was sufficiently sensitive to quantify the low concentrations (0.3mg CN?/L) of CN? found in ruminant blood. In goats infused with KCN, the CN? concentration in blood was time-dependent and continued rising during infusion, gradually declining after infusion ceased after 1 h. Blood CN? levels showed a time-dependent increase in all experimental rabbits with the maximum concentration (1.34 mg/L) at day 40. The highest blood CN? concentrations were found in cattle, followed by goats and buffalo, with the lowest in sheep. When blood CN? levels of these species were compared statistically, a non-significant difference was observed. A non-significant difference was found between males and females. No relationship was observed between blood CN? levels and age of the animal for any species. Buffalo, cattle, sheep, and goats allowed to graze showed significantly higher blood CN? levels than animals kept in a stall feeding system. Animals grazing on jumbo grass were found to have significantly higher blood CN? levels than those fed S. bicolor or S. sudanese with a non-significant difference found between animals fed S. bicolor or S. sudanese. No CN? was detected in blood samples of animals fed maize. Hydroxocobalamine was found to be a significantly more effective CN? antidote than garlic, STS, SNT plus garlic extract, or SNT and STS, either alone or in combination. A combination of SNT and garlic extract was the second most effective CN? antidote. The efficacy of garlic alone was significantly higher than STS or SNT in combination with STS. The efficacy of combined SNT and STS was superior to STS alone in treating rabbits with CN? toxicity. No rabbits in any group demonstrated gross deviation from the normal organ structure. The activities of serum ALT, AST, ALP, and LDH enzymes, as well as serum bilirubin, were significantly increased in CN? treated rabbits compared to controls. Severe hepatocyte vacuolation and degeneration were present in liver of rabbits in the CN? treated group. Liver of rabbits in the control group showed normal morphological patterns. The concentrations of serum urea, uric acid and creatinine were significantly higher in CN? treated rabbits than in control group. Urinary thiocyanate levels were also significantly higher in the CN? group than in controls. Kidneys of rabbits in the CN? group demonstrated severe glomerular and tubular necrosis and congestion. Pyknotic nuclei were present in tubular epithelial cells, whereas a normal histological pattern was observed in kidneys of rabbits in control group. Serum T3 and T4 levels were significantly lower in the CN? group compared to controls. A non-significant difference in blood glucose levels was recorded between rabbits in control and CN? treated groups, and histological examination of pancreas revealed no microscopic lesions. No significant differences were observed in serum cholesterol levels of CN? and control group rabbits. The serum albumin and total protein concentration in CN? treated rabbits were significantly lower than in the control group. Heart of rabbits in both CN? and control group did not show histopathological changes under microscopic examination. The erythrocyte count, the hemoglobin concentration, packed cell volume and mean corpuscular hemoglobin were all found to be significantly lower in blood of CN? treated rabbits than control group. The mean corpuscular hemoglobin concentration was significantly higher in the CN? group rabbits than control group animals. On the other hand, the difference in leukocyte count, differential leukocyte count and platelets were non-significant. The difference in total and daily feed consumption between CN? and control group rabbits was non-significant, whereas the feed efficiency of rabbits in the control group was significantly higher than for rabbits in CN? fed group. The net weight gain of rabbits in the control group was significantly higher than in the CN? fed group. Cyanide infusion to pigs resulted in toxic levels of blood CN? accompanied by lactic acidosis. In addition, there was a progressive increase in cardiac output, Venous oxygen saturation, heart rate, elevation of central venous pressure and pulmonary artery blood pressure. Skeletal muscle rSO2 progressively and significantly decreased with increasing lactate and CN? levels. However, there was no significant change in brain rSO2.
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Alleviation Of Cyclic Heat Stress In Broilers By Dietary Supplementation Of Mannan-Oligosaccharides And Lactobacillus-based Probiotic
Material type: Book ; Format:
Publisher: 2011 Dissertation note: The antiviral activity of plants Silybum marianum (seeds), Chenopodium album (whole plant) and Nigella sativa (seeds) were evaluated against Peste des petitis ruminants virus (PPRV) and Foot and Mouth Disease virus (FMDV) in this study. Methanolic extraction of these plants was done by using Soxhlet apparatus and extracts were dried by using rotary evaporator. Six dilutions of each extracts 100, SO, 2S, 12.S, 6.2S, 3.12~g/ml were made in distilled water. Vero cells were infected by PPRV and BHK-21 by FMDV respectively. The herbal extracts assays of antiviral and cytotoxic were carried out in cell culture plates. Each well of 96 well cell culture plate were seeded with 104cell/ml of cell suspension. Cell counting was performed by hemocytometeric method. Positive and negative controls for antiviral and cytotoxic assay were also used, incubated the 96 well cell culture plates at 37°C for 4 days. After this incubation, MTT [3-(4,S-dimethylthiazol-2-yl)- 2,S-diphenyltetrazolium bromide] colorimetric assay were used for the determination of their quantification. Endpoint of this assay was considered in terms of cell survival percentage. Results were compared for qualitative variables using Chi-square technique and quantitative variables by linear regression analysis. 1 OO~g/ml and SOIlg/ml concentrations of Chenopodium album showed cell survival percentages of 87.9% and 86% respectively in PPRV and all six test dilutions of same plant showed no cytotoxicity for Vero cells. IOuug/ml and
SO~g/ml concentrations of Chenopodium album showed cell survival percentages of 88.5% and 87.2% respectively in FMDV and all six test dilutions of same plant showed no cytotoxicity for BHK-21 cells. Two concentrations of Nigella sativa 50!J. glml and 25!J. glml showed prominent cell s urvival of 85% and 84% respectively in PPRV and only one concentrations l Ouug/ml were found cytotoxic.Two concentrations of Nigella sativa 50uglml and 25!J.glml showed prominent cell survival of 79% and 77% respectively in FMDV and only one concentrations IOuug/ml were found cytotoxic. Only IOuug/ml of Silybum marianum has shown cytotoxicity and 50!J.glml and 25!J.glml shown prominent antiviral activity 91% and 85% respectively in PPRV. In FMDV l Otlug/ml of Silybum marianum has shown cytotoxicity and 50!J.glml and 25!J.g/ml shown prominent antiviral activity 93% and 91 % respectively. The results of present study are helpful in the treatment of Peste des petitis ruminants and Foot and Mouth Disease.
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Sero Epidemiology Transmission Dynamics And Hematological Studies On Neospora Caninum In Dairy Buffaloes
Material type: Book ; Format:
Publisher: 2011 Dissertation note: The current study was carried out in two districts of the Punjab province (pakistan),
viz. Lahore and Narowallocated;on the eastern part of the province bordering Indian Punjab
and the first study assessing the epidemiology of N caninum in dairy buffalo in Pakistan.
Overall, the true sercprevalence of Neospora caninum in dairy buffalo (assessed by using
cELISA validated for buffalo by using IFAT as reference test) of the two districts was 39.4 %
with no significant difference in the prevalence of N caninum antibodies between male
(29.5%) and female (41.5%) dairy buffalo. However, in the Lahore district alone, the
prevalence in the female (46.1 %) buffalo was significantly higher than in their male (23.8%)
counterparts. In contrast, in Narowal district, the prevalence in male (42.4%) buffalo was
higher than females (32.2%) but this difference was not statistically different. The other risk
factors evaluated in regards to the prevalence of N caninum in buffalo were similar overall in
both the districts; these included age-specific, season-specific, pregnancy-specific, parity-
specific, abortion-specific and dog contact-specific prevalence. The difference in sero-
prevalence was not significant amongst all the ages but was significant between the younger
" 3yrs.) and adult (> 3yrs.) age buffalo. The adult buffalo showed a higher prevalence of N
caninum antibodies compared with younger buffalo. Anyhow, the age group of buffalo >3-5
yrs. showed consistently the highest prevalence of N caninum antibodies overall and in both
the districts in the current study. The summer season was the one in which buffalo of the two
districts and overall showed the highest prevalence, followed by high prevalence in autumn; while
the lowest prevalence of N caninum antibodies was observed in buffalo sampled during the
winter. The prevalence in the higher parities buffalo was significantly higher than the lower parity
and pregnant heifers. The prevalence in pregnant dams was significantly higher than in non-
pregnant buffalo in the herds in different areas of the two districts.
Similarly, aborting buffalo in regular contact with dogs- at the dairy properties showed significantly
higher prevalence of N. caninum antibodies than non-aborting buffalo without the contact of dog.
The serological analysis-of dogs (using cELIS A) present at some dairy buffalo
properties generally showed high prevalence of N. caninum; antibodies at the dairy properties
where the buffalo were in regular contact with dogs compared with farm premises and their
buffalo without any contact with dogs at the farm. This signifies the roleof the dogs in the
. transmission of N. caninum oocysts and infection to buffalo (intermediate host) as the isolates
of dog and bovine origin has been reported to be identical (Stenlund et al., 1997).
The analysis of milk samples from the dairy buffalo. also tested for serum antibodies
using iscom (immune stimulating complex) ELISA and showed a good level of agreement
(Kappa ratio=O.567) between the performance of two tests (iscom ELISA versus cELISA).
However, the cELIS A (VMRD, Pullman, W A) showed higher sero-positivity compared to
iscom ELISA (SV ANOV A, Uppsala, Sweden). Therefore, the use of either ELISA used on
milk for evaluating the prevalence of N. caninum antibodies in buffalo can give good results
. for lactating buffalo, but cELIS A appeared to have superior sensitivity in our study. The use
of iscom ELISA offers advantages regarding ease of sampling and repeated screening of
lactating herds but with little lower sensitivity than serum antibody cELISA.
The co-existence of N. caninum with Br. abortus, another important abortifacient
pathogen in buffalo, is quite significant as the probability of such infectious abortions is
increased in the susceptible herds resulting in more economic losses compared to those
caused by single pathogen. The co-existence of Br. abortus in N. caninum seropositive
buffalo in the current study was 13.2%, while the overall prevalence of Br. abortus antibodies
in buffalo of two districts was 12.2%. The prevalence trend of Bt: abortus was related to age,
though not exclusively; as prevalence continued to increase with age except in the age groups
of 6-7 yrs, and> 7 yrs. old buffalo. The higher prevalence in the adult age groups indicates
the association of Br. abortus with development of the reproductive system of the host
(buffalo). It also suggests the increase in the exposure of the buffalo to the pathogen with
increase.in age to some good extent. However, this prevalence was statistically not signific~
in all the age groups of buffalo. The prevalence of co-existence was highest (18.5%) in -,
buffalo >3-5 yrs. old followed by 18.2% in buffalo >7 yrs. age. Howe er, statistically, the
difference in the prevalence of co-existing antibodies against N caninum and Br. abortus in
buffalo was not significant.
Hematological analysis of the N caninum sere-positive and sero-negative dairy
buffalo for different haematological determinants, such as total leukocytic counts (TLC),
differential leukocytic counts (DLC), RBCs, haemoglobin, haematocrit, platelets and blood
plasma glucose level showed no difference in sero-positive and sero-negative two groups
particularly, regarding the effect of sero-positivity in buffalo. Albeit, some interactions such
as season x N caninum seropositivity showed significantly lower monocyte counts and high
blood glucose level in N caninum sere-positive buffalo compared with sero-negative buffalo.
Overall, in both groups no significant changes were observed in the seropositive buffalo
compare to sero-negative ones. Some interaction have independently affected certain
haematological parameters although, the animals were not seropositive. This suggests the
effect of some environmental factors and physiology states of the buffalo on its haematological
dynamics. These effects included the effect of lactation on the lymphocyte count in the lactating
and non-lactating buffalo, where lymphocyte counts were higher in non-lactating compared with
lactating buffalo most possibly due to the recruitment of lymphocytes in the udder for excretion in
milk. Similarly, blood haemoglobin was lower in the hot months and packed cell volume was
higher in the cooler months of the year in the buffalo.
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Epidemiology, Serodiagnosis And Chemotherapy Of Anaplasmosis In Cattle
Material type: Book ; Format:
; Literary form:
Publisher: 2011 Dissertation note: Anaplasmosis is globally distributed tick-borne disease of livestock with great
economic importance in cattle industry. The current project was designed to estimate the
prevalence of anaplasmosis, ticks and risk factors associated with seroprevalence of
Anaplasma marginale among cattle in Sargodha, Khushab and Rawalpindi districts,
Punjab, Pakistan. Moreover, haematological changes in A. marginale infected cattle and
efficacy of chemosterilization regimens were evaluated using locally available drugs for
the elimination of adult naturally infected carrier cattle.
A total of 1050 blood, serum and tick specimens were collected from randomly
selected small holders (n=90) and private livestock farms (n= 12) using multistage cluster
random sampling technique. A total of 30 union councils, 34 cattle farms (30 small
holders and 4 livestock farms) and 350 cattle were selected as primary, secondary and
elementary sampling units from each district. Sampling unit was indigenous and
crossbred cattle of both the sexes.
Microscopic examination of the Giemsa stained blood mears revealed an overall
prevalence of blood parasites as 21.14%. Anaplasma marginale was the highe t prevalent
(5.81 %) haernoparasite of cattle followed in order by Theileria sp. (5.14%) and Babesia
bigemina (4.76%), respectively. Crossbred cattle were more susceptible to TBDs as
compared to the indigenous cattle. Highest prevalence of TBDs was recorded in summer.
The prevalence of tick-transmitted diseases was higher in small holders (31.3%) than
private livestock farms (17.5%). Chi square analysis indicated a significant association
(P<0.05) among indigenous and crossbred cattle to selected TBDs. Wherea. non-
significant association between different age groups, seasons, sex and farm sizes were
The overall seroprevalence of Anaplasma marginale in cattle using cELlSA was
31.05%. Seroprevalence was higher in crossbred cattle of more than 4 years of age and
there was a significant (P<O.OO I) association between different age groups and breed. The
seroprevalence was significantly (P<0.05) higher in summer season in Sargodha and
Khushab districts. Moreover, the seroprevalence was significantly higher in small holders
in all study districts.
The epidemiological data and relevant information regarding area, host and farm
management factors were collected on a questionnaire through interview from each dairy
farmer, attendant or manager from September, 2009 to August, 20 10. Multivariate
analysis of risk factors revealed that cattle of more than 4 year of age (OR=5.42), heavy
tick infested (OR =2.10), crossbred (OR = 1.59) cattle were significantly at higher risk for
seroprevalence to Anaplasma marginale. Presence of Rhipicephalus (Boophilus)
microplus (OR=3.70), use of ivermectin (OR=3.97), moderate interval of acaricide
frequency (OR= 16.50), stall feeding (OR=4.90) and use of unhygienic needles
(OR=24.00) were significantly associated with seroprevalence to Anaplasma marginale
in cattle (P<0.05). The Sargodha district was at higher risk (OR = 1.81) as compared to
Khushab and Rawalpindi.
The tick species identified from cattle were Hyalomma anatolicum anatolicum,
Rhipicephalus (Boophilus) microplus, Rhipicephalus sanguine us, Rhipicephalus
(Boophilus) annulatus and Haemaphysalis sp. The overall prevalence of tick infestation
among cattle was 54.76%. The highest prevalence (57.71%) of cattle tick infestation was tick infested sites in cattle followed by dewlap (92%), inner thighs (90%), neck & back (54%), tail (26%), ears (13%), around eyes (10%), flanks (4%) and legs (2%).
The haematological changes were studied at different levels of parasitaemia " 7%, >7-15% and> 15%) in Anaplasma marginale infected Sahiwal and crossbred cattle. There was a significant difference (P<O.OS) among total erythrocyte count (TEC), total leukocyte count (TLC), haemoglobin (Hb), packed cell volume (PCV), mean corpuscular
haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC) at different levels of rickettsemia in both breeds. ignificant difference (P<O.OS) was noticed among RBCs, PCV and MCH blood parameters between Sahiwal and crossbred cattle.
A total of sixty Anaplasma marginale seropositive adult Sahiwal cattle were selected having their ages between 3-4 years ranging in weight from 246-341 kg. The animals were divided in four groups designated as OXY -group-I, E RO-group-II, IMC- group-III and control-group-IV, comprising IS animals each. The seropositive animals received oxytetracycline (22 mglkg IV once in a day for five days), enrofloxacin (S mglkg IV once in a day for five days) or imidocarb (S mglkg 1M twice, 7 days apart). Re ult of chemosterilization study indicated that oxytetracycline 13/1S (86.67%) and irnidocarb
dipropionate II/IS (73.33%) eliminated Anaplasma marginale infection in adult naturally infected carrier cattle on S6th day. The carrier clearance was confirmed by cELISA followed by subinoculation of blood in seronegative splenectomized calves.
It was concluded that TTBDs are widely distributed in Punjab, Pakistan. Host. management and area factors are involved with the seroprevalence of Anaplasma marginale in cattle. Haemolytic anaemia is the major haematological finding of Anaplasma marginale in cattle. Oxytetracycline is more effective and safe In chemosterilization of persistent Anaplasma marginale infection in cattle. There is a need for country wide epidemiological studies on ticks and TBDs using advanced serological
and molecular techniques. Moreover, the identification of the potential vector of anaplasmosis should be required for the effective prevention and control of anaplasmosis in Pakistan.
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Isolation, Characterization And Pathogenesis Of Capripox Virus
Material type: Book ; Format:
; Nature of contents: ; Literary form: Publisher: 2010 Dissertation note: Goat pox is the most important pox diseases of livestock and it usually
causes huge economic losses. The economic losses occur in terms of mortality,
reduced productivity and lower quality of wool and leather. The clinical
manifestations of the disease include high temperature, lesions skin in the form of
macules, papules, vesicles, pustule and scabs on hairless areas of the body. The
disease is highly contagious having high morbidity and mortality in the infected
herds. The present study was conducted to document the prevalence of goat pox
disease in the different regions of Punjab. The study was based on clinical
manifestation of the disease in various collecting spots including slaughter houses,
cattle and hide markets and tanneries. The prevalence of goat pox at slaughter
houses in different regions was 9.93% in arid region followed by 8.69% and 7% in
southern and northern irrigated regions respectively. The prevalence of pox disease
in sheep was highest (8.54%) in the northern irrigated region, 7.69% and 6.62% in
arid and southern irrigated regions respectively.
The prevalence of pox recorded in the hide markets shows a trend of high
presence 7.29% in arid region followed by 6.22% and 3.84% in southern and
northern irrigated regions. Whereas in sheep the overall prevalence was 0.51 %,
4.44% and 1.66% in northern irrigated, arid and southern irrigated regions.
In tanneries the pox lesions were identified on the basis of method as
adopted in hide markets. The overall prevalence of pox in goat was 3.96%, 4.06%
and 4.09% while in sheep 9.58%, 2.41 % and 10% in northern irrigated, arid and
southern irrigated regions.
The overall prevalence of pox disease in goat was 5%, 5.79% and 5.34% in
Northern irrigated, arid and southern irrigated regions respectively. Where as in
sheep, pox was 3.133%, 4.11 % and 2.67% in Northern irrigated, arid and southern
irrigated regions respectively. The highest trend of incidence of disease was present
in the arid regions followed by southern and northern regions. The slaughter houses
shows high incidence of disease as compared to cattle and hide market and
tanneries. The result was significant (P<0.05) among the regions and samples
A total of 100 samples consisting of 55 scabs and 45 skin tissues were
randomly selected from the different collecting spots of the three regions. The scabs
and skin tissue samples were processed on dehydrated minimum essential media
tor virus isolation. The virus was isolated on Vero cell line culture and its
characteristics were observed on the basis of specific cytopathic effects. All 55 scab
samples consisting 20 from cattle markets, 20 from slaughter house and 15 from
hide market and tannery were tested through cell culture. The cell culture positive
result for scabs was 60% cattle markets, 20% hide market and tannery and 40%
All 45 skin tissue samples including 5 from cattle markets and tannery, 20
from hide market and 20 from slaughter house were subjected to virus isolation on
Vero cell line. The cell culture positive result for skin tissue samples was 100% cattle
markets, 30% hide market and tannery and 60% slaughter house. In this way the
total cell culture result for scabs and skin tissue samples from all areas become
41.82% and 51.11 % respectively.
The isolated virus was confirmed through peR. All the collected samples
were also analyzed through peR in order to compare the two techniques for disease
diagnosis. Out of 40 samples from slaughter houses 18 scabs and 15 tissues
sample were positive through peR with 82.5%. Out of 25 samples collected from
cattle markets consisting of 20 scabs and 5 skin tissues, 17 of scabs and 5 skin
tissues were positive with 92%.
Similarly a total of 35 samples out of which 15 were scabs and 20 were skin
tissues collected from hide markets and tanneries. The peR of 7 scabs and 14 skin
tissues was positive with 60%. In this way the total peR result for scabs and skin
tissue from all areas was 42% and 34% respectively.
In the 3rd study of the present project the isolated virus was inoculated in to
experimental animal to study the detail pathogenesis. The disease followed the
same pattern as in the natural outbreak. But however the routes of inoculation affect
the severity of the disease. During the study the diseased animals were periodically
slaughter at weekly interval after the appearance of 1 st clinical signs. The detailed
lesions were observed in different visceral organs and the tissues were collected
and preserved in 10% formalin. The tissues were processed for histopathology and
immunohistochemical examination. The IHC was successfully optimized for the
detection of viral antigen in the tissues of skin, lung and lymph nodes.
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Bat Biodiversity (Vespertilioniformes: Order Chiroptera) In Some Tropical And Arid-Subtropical Regions Of Pakistan
Material type: Book ; Format:
; Literary form:
Publisher: 2011 Dissertation note: The present study was conducted from June 2009 to May 2011 in those arid subtropical and tropical regions of Pakistan which included less pronounced monsoon influenced areas of the Salt Range, the Upper Indus Plains and the sand dune areas typified by the Cholistan. Bat surveys were conducted in two protected areas i.e. the Margallah Hills National Park (SA1) and the Chinji National Park (SA2) that were located in the arid subtropical region and in another, the Lal Suhanara National Park (SA3), situated in the tropical sand dune region of the Upper Indus Plains. In addition, bat samples were also collected from Gujranwala, Lahore, Tob Tek Singh and Kasur districts (SA4). These sub-areas were selected to maximize the chances of capture of as many bat species inhabiting arid-subtropical and tropical habitats of Pakistan as possible.
A total of 182 bats belonging to twelve species were recorded. These included R. blasii (Family Rhinolophidae), R. hardwickii (Family Rhinopomatidae), Taphozous nudiventris and T. perforatus (Family Emballonuridae), Scotoecus pallidus, Scotophilus heathii, S. kuhlii, Pipistrellus ceylonicus, P. javanicus, P. pipistrellus, P. tenuis and Hypsugo savii (Vespertilionidae). Rhinolphous blasii was captured only from SA1, R. hardwickii and S. pallidus from SA3 and P. tenuis from SA1. Taphozous nudiventris and T. perforatus were captured from SA1 and SA3, S. kuhlii and P. ceylonicus from SA1 and SA4, H. savii from SA1 and SA2 and P. javanicus from SA1 and SA2. Scotophilus heathii and P. pipistrellus were recroded throughout the study area.
Maximum bat activity was recorded in spring (n = 65) that was followed by summer ( n = 61), autumn (n = 32) and winter (n = 24). Rhinolophus blasii and S. pallidus were recorded only during winter, R. hardwickii and P. tenuis during autumn, while S. kuhlii was recorded only during summer. Taphozous nudiventris and T. perforatus were captured during summer and autumn. Pipistrellus pipistrellus was recorded during autumn, spring and winter while S. heathii was captured throughout the year.
Although the netting effort was the same, the number of bats captured from the SAs was different. A total of 72 bats were recorded from SA1, 52 from SA4, 43 from Lal SA3 and 15 from SA2. The dominance was highest for SA2 and lowest for SA1. Both Shannon and Simpson indices show that the diversity was the highest at SA1 followed by SA3, SA4 and SA2. Evenness was found to be highest at SA4 followed by SA3, SA2 and SA1.
The mean head and body length of three Rhinolophus blasii was 39.33 mm ± 0.577 (SD) forearm length was 40.17 mm ± 1.155 (SD) and the tail length was 19.23 mm ± 1.940 (SD). The greatest skull length of a single R. blasii was 17.22 mm and mandible length was 11.80 mm. The baculum of a single R. blasii sample was 3.5 mm long.
The mean head and body length of two Rhinopoma hardwickii 66.00 mm ± 5.657 (SD). The mean forearm length was 54.00 mm ± 0.0 (SD). The tail length was 59.00 mm ± 2.828 (SD). The greatest skull length was 19.68 mm ± 0.108 (SD), and the length of mandible was 11.28 mm ± 1.652. The baculum of single R. hardwickii was 1.1 mm long.
The mean head and body length of twenty six Taphozous nudiventris was 86.87 mm ± 5.556 (SD) and the tail length was 27.57 mm ± 12.187 (SD). The greatest skull length was 26.16 mm ± 0.323 (SD) and the length of mandible was 17.53 mm ± 1.149 (SD). The mean total baculum length of the two specimens was 0.58 mm ± 0.017 (SD).
The head and body length of four T. perforatus was measured as 84.30 mm ± 5.450 (SD) long. The forearm was 64.30 mm ± 3.457 (SD) long and the length of tail was 22.10 mm ± 2.702 (SD). The greatest length of skull was 22.24 mm and the length of mandible was recorded as 16.25 mm. The total length of a single T. perforatus was measured as 0.69 mm. The head and body length of fifty three Scotophilus heathii was 79.46 mm ± 6.941 (SD). The mean forearm length was 58.69 mm ± 2.929 (SD) and the tail length was 55.00 mm ± 7.360 (SD). The greatest length of skull was 21.39 mm ± 1.378 (SD) and the length of mandible was recorded as 16.08 mm ± 0.882 (SD). Mean total bacular length of ten S. heathii was measured 1.76 mm ± 0.150 (SD).
The mean head and body length of five specimens of S. kuhlii was 72.10 mm ± 8.096 (SD). The forearm was 49.40 mm ± 3.03 (SD) long and the length of tail was 42.40 mm ± 4.04 (SD). The greatest length of the skull was 18.98 mm ± 0.613 (SD) and the mandible length was 14.41 mm ± 1.173 (SD). The total length of the baculum of a single S. kuhlii was 1.74 mm.
The head and body length of two Scotoecus pallidus was 56.50 mm ± 3.536 (SD). The forearm was 35.50 mm ± 0.707 (SD) long and the length of the tail was 35.50 mm ± 3.536 (SD). The greatest length of skull was 15.46 mm ± 0.449 (SD) and mandible length was measured 9.64 mm ± 2.425 (SD). The total length of the baculum of a single S. pallidus captured from SA3 was 5.0 mm.
The mean head and body length of twenty two Pipistrellus ceylonicus was 63.60 mm ± 7.486 (SD). The length of forearm was 29.92 mm ± 2.492 (SD) and tail length was 25.68 mm ± 3.442 (SD). The greatest length of the skull was 10.76 mm ± 0.257 (SD) and the length of mandible was 9.28 mm ± 3.956 (SD), respectively. Mean total length of the bacula of four P. ceylonicus was 3.66 mm ± 1.190 (SD).
Mean head and body length of the ten P. javanicus was 52.00 mm ± 2.712 (SD). The forearm was 35.13 mm ± 1.996 (SD) long and the length of the tail was 30.38 mm ± 5.236 (SD). The greatest skull length was 13.01 mm ± 4.546 (SD) and the length of mandible was 10.29 mm ± 1.679 (SD). The mean total length of the four bacula was 3.57 mm ± 0.860 (SD).
The head and length of fifty two P. pipistrellus was 39.33 mm ± 2.690 (SD). The forearm was 28.23 mm ± 1.264 (SD) long and the length of the tail was 25.86 mm ± 3.396 (SD). The greatest length of skull was 11.04 mm ± 0.342 (SD) and the length of mandible was 7.87 mm ± 0.802 (SD). The mean total length of the eleven bacula of P. pistrellus was 3.19 mm ± 0.421 (SD).
Only two specimens of P. tenuis were captured from SA1. The head and body length of these specimens was 35.00 mm ± 2.828 (SD). The forearm length was 28.00±0.707 while the length of the tail was 22.25 mm ± 3.182 (SD). The greatest length of the skull was 10.19 mm. and the mandible length was 7.82 mm. The total bacular length was 2.79.
The head and body length of the two Hypsugo savii was 55.50 mm ± 19.092 (SD). The forearm was 36.75 mm ± 3.889 (SD) long while the length of the tail was 33.50 mm ±6.364 (SD). The greatest length of the skull was 11.18 mm and the length of mandible was 7.08 mm. The total bacular length of a single H. savii was 2.67 mm.
The echolocation calls of bats of Pakistan have never been recorded and thus the accuracy of species identification on the basis of their calls remains a bit doubtful.
Availability: Items available for loan: UVAS Library [ Call number: 1374,T] (1).
Effects Of Stair-Step Nutrition Regimen On Growth Rate, Nutrien Utilization And Pubertal Development In Nili-Ravi
Material type: Book ; Format:
; Literary form:
Publisher: 2011 Dissertation note: Under this study, effect of stair-step nutritional regimen compared to the standard NRC recommended energy levels on growth rate, nutrient utilization, some selected blood metabolites, pubertal age, conception rate and economic analysis in ili- Ravi buffalo heifers were measured. Study lasted for 18 months during the years 2008- 20 I O. Twenty-two heifers, 6-8 month old, 98.57±5.07 kg average ody weight were divided into two equal groups and randomly assigned either control or stair-step nutritional regimen (SSNR) diets. The SSNR was designed in three phase program each having 6 months duration i.e., postweaning (7 to 13 month age), repubertal (13 to 19 month age) and pubertal/breeding (19 to 25 month age). In each phase, the treatment group during step 1, was fed on low energy diet (80% ME of NRC) for 4 months followed by high energy diet (120% ME ofNRC) for 2 months in step 2. The heifers in ontrol group were fed according to NRC (200 I) requirements of Holstein Friesian heifers continuously for 6 months. For both the groups individual feeding was carried out. Daily feed intake and fortnightly fasting weights were recorded. Nutrients digestibility and N balance trials were conducted during last week of each step during each phase. Blood samples were collected at the end of each low or high energy diets for blood metabolites analysis. Oestrus detection was done with the help of a teaser bull at age of 15-16 months. Transrectal ultrasonography was done to assess uterus and ovarian structures development. Measured blood serum progesterone concentration collected every 10 days interval at 09.00-10.00 hours during 18-20 months age by ELISA using commercial kit. The age and live weight at onset of puberty was recorded when heifer tood to be mounted by the bull first time in her life. The heifers detected in oestrus were bred by natural mating at approximately 12-15 hours of the onset of oestrus activity. Heifers not returning to oestrus were examined for pregnancy diagnosis through rectal alpation of uterus at 70-90 days post breeding. Data of feed onsumption during postweaning, prepubertal and pubertallbreeding phases were used to calculate the feed cost used per kg gain between the SSNR and control heifers.
During postweaning phase, heifers fed SSNR low energy diet (2.03 Meal/kg) ained significantly (P<O.OS) lower daily weights than those fed control diet (2.SS Meal/kg), When heifers fed high energy diet (3.01 Meal/kg), daily weight gain was significantly (P<O.O 1) higher in SSNR compared to control. Average dry matter intake (DMI) was similar (P>O.OS) between the heifers of two groups. Feed conversion ratio (FCR) was poorer (P<O.OS) in SSNR heifers fed low energy diet compared to those fed control diet. But on high energy diet FCR was better (P<O.OS) in SSNR compared to control group.
During prepubertal phase, there was no difference (P>O.OS) in weight gain between the heifers fed SSNR low energy diet (1.89 Meal/kg) and control diet (2.3S Meal/kg). But on high energy diet (2.80 Meal/kg) weight gain was higher (P<O.OS) in SSNR compared to control group. Average dry matter intake (DMI) was similar (P>O.OS) between the heifers of two groups. On low energy diet there was no difference (P>O.OS) in FCR between the two groups. But on high energy diet FCR was significantly (P<O.OS) better in SSNR compared to control group. Average DMI in heifers of both groups was similar (P>O.OS). During pubertal/breeding phase, similar trend of weight gain, DMI and FCR was found in SSNR versus control group as reported in prepubertal phase.
Intake of DM, organic matter (OM) and crude protein (CP) as percent body weight were statistically non-significant (P>O.OS) differet between the SSNR versus ontrol groups during all phases. Metabolizable energy (ME) consumption was significantly P<O.OS) lower in SSNR group fed low energy diet than the heifers fed control diet. But ME consumption was significantly (P<O.O 1) increased in SSNR group fed high energy diet than control group. Similar, trend of ME consumption was observed in heifers fed SSNR (either low or high energy) and control diets during prepubertal and pubertal phases. Water to dry matter intake ratio in heifers during postweaning, prepubertal and pubertal phases were statistically similar (P>O.OS).
In all phases, apparent DM and OM digestibility did not differ (P>0.05) between the heifers fed SSNR (either low or high energy) and control diets. Neutral detergent fibre (NDF) digestibility was higher (P<0.05) when SSNR heifers fed low energy diet, but on high energy diet NDF digestibility was significantly (P<0.05) lower compared to control, respectively, during all phases with the exception of step I in the prepubertal phase and step 2 in pubertal phase where the differences were non-significant (P>0.05) between the groups. Acid detergent fibre (ADF) digestibility with SSNR low energy diet was significantly (P<0.05) higher than the heifers fed control diets during three phases. But on high energy diet, ADF digestibility was not different (P>0.05) between the two groups. Also N intake was not different (P>0.05) between the heifers fed SSNR (either low or high energy) diets and control diets, respectively, with the exception of step 2 in the postweaning phase when the control group showed a significant (P<0.05) increase in intake of N compared to the SSNR group. Faecal N as well as Urinary N losses in heifers fed SSNR (either low or high energy) versus control diets did not differ significantly (P>0.05).
All heifers have shown haematological values which are almost similar in heifers of two groups. Except total cholesterol, concentration of urea N, glucose and macro minerals in serum did not differ between the two groups.
There was no significant (P>0.05) differences in age and weight at onset of puberty and number of services per conception between the two groups. Pregnancy rate in heifers fed on SSNR diet was 50% while on control diet was 57%. Fifty percent of heifer fed SSNR and 60% of heifers fed control diet as per NRC requirement had serum progesterone concentrations> 1.0 ng/ml in two samples collected 10 days apart before reaching puberty.
The overall feed costs incurred (42660.88 vs 44509.96 Rs./animal) on SSNR heifers was significantly (P<0.05) less than the control heifers fed according to NRC recommendations from weaning to breeding age.
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Prevalence, Serodiagnosis And Zoonotic Importance Of Hydatidosis In Small Ruminants And Humans
Material type: Book ; Format:
; Literary form:
Publisher: 2011 Dissertation note: Hydatidosis is a world-wide zoonotic parasitic disease which is caused by the larval stage of tape worm Echinococcus granulosus and E. multilocularis. It is highly endemic in some regions of the world. Keeping in view the importance of the disease, the present study was conducted to record the age, sex, species and season wise prevalence of hydatidosis in small ruminants. The study also includes comparison of the two serological tests used for the diagnosis of this disease and efficacy of mebendazole was aIso evaluated. Prevalence and organ specificity of hydatidosis was studied in 2400 sheep and 2400 goats of different age and sexes. The overall prevalence of hydatidosis in sheep and goats was 8.25 and 6.21 percent respectively. Sex wise prevalence indicated that it was 9.85 and 7.85 percent in female and male sheep respectively whereas in female and male goats it was 5.83 and 7.23 percent respectively. A reasonably higher prevalence of hydatidosis was observed in female animals as compared with males. In adult animals hydatid cysts were present in 11.38 and 7.77 percent in sheep and goatsrespectively while in lambs and kids it was 1.79 and 1.06 percent respectively. Statistically no significant difference was observed in any season through out the year in both the species. In the present study, lungs were found to be most commonly infected organ in sheep andliver in goats. Out of 198 infected sheep, 45.45 percent had cysts in lungs, 33.84 percentin liver, 10.10 percent in both liver and lungs, 2.53 percent in spleen, 2.02 percent in heart and 6.06 percent in abdomen and thoracic cavity whereas in goats organs specificity was 34.23, 40.27, 16.78, 0.67, 8.05 percent respectively for lungs, liver, lungs & liver both, spleen and abdomen and thoracic cavity. A total of 60 cysts in sheep were examined for fertility and it was found that out of 18 cysts of less than two centimeter size, maximum (50 percent) were suppurative orcalcified and minimum (16.7 percent) were fertile in nature. Of 30 cysts of 2-4 em size,
40 percent were suppurative and 40 percent were fertile and from 12 cysts of more than
four centimeter size, maximum (66.7 percent) were fertile and only 16.7 percent were
sterile in nature. In goats, of 15 cysts of less than 2cm size, maximum (46. 7percent) were
suppurative, and minimum (13.3 percent) were fertile. Of 7 cysts, belonging to the group
of more than four centimeter seize, 71.4 percent were fertile in nature and only 14.3
percent were sterile. Overall fertility rate in sheep and goats was 38.33 and 36.96 percent
respectively whereas overall sterility rate was 23.3 and 32.60 percent in sheep and goats
In sheep, total number of protoscolices found in 23 cysts was 936 of which 72.65 percent
were fertile in nature. The mean of total proto scolices was 40.70 with standard deviation
of 23.05 whereas the mean number of viable protoscolices was 29.57 percent with a
standard deviation of 18.92. In goats, total number of protoscolices observed in 17 cysts
was 719 whereas only 52.71 percent were fertile in nature. The mean of total
protoscolices was 42.30 with a standard deviation of 17.13 whereas mean of viable
protoscolices was 22.30 with standard deviation of 17.10.
Blood samples of 40 positive and 40 negative sheep for hydatid disease were collected
from slaughter house and serum was separated. ELISA and IHA test were applied on
these samples. The sensitivity, specificity, efficacy, positive predictive value and negative
predictive value of ELISA were 92.5 percent each whereas these values for IHA were 80
percent, 97.5 percent, 88.75 percent, 96.96 percent and 82.98 percent respectively. In
goats, blood samples from 40 positive and 40 negative cases of hydatidosis were
collected from slaughter house before slaughtering and ELISA and IHA were applied on
all of these samples. The sensitivity and specificity of ELISA was 90 percent and 95
percent respectively whereas the sensitivity and specificity of IHA was 75 percent and
97.5 percent respectively.
A total of 12 i.e. 6 sheep and 6 goats (healthy) were purchased and kept at Ravi campus
Pattoki, University of Veterinary and Animal Sciences, Lahore. These were divided into
experimental (4 sheep and 4 goats) and control group (two sheep and two goats). In
experimental group, 100 eggs of Echinococcus granulosus were given orally to all
animals and then mebendazole was given to two sheep and two goats on zero day and
then after two months intervals. Blood samples were collected from all animals on zero
day, 90 days and 180th day and serum was separated. ELISA and IHA were applied to all
samples. Mebendazole was not found completely effective and ELISA detected the
infection earlier than IHA. On postmortem examination, multiple cysts were found in all
animals of group A and no cyst was found in group B.
Eighty blood samples of humans i.e. 40 positive and 40 negative for hydatidosis were
coilected, serum was separated and ELISA and IHA test were applied to all the samples.
The sensitivity and specificity of ELISA was 95 percent and 97.5 percent respectively
whereas the sensitivity and specificity of IHA was 82.5 percent and 97.5 percent
The present study will be helpful in disseminating the informations regarding the
prevalence, zoonotic importance, effect of mebendazole in animals and the use of
immunodiagnostic tests for the diagnosis of hydatidosis in small ruminants and human
Availability: Items available for loan: UVAS Library [ Call number: 1405,T] (1).
Comparison Of Diagnostic Approaches For The Detection Of Bovine Viral Diarrhea Persistency In Dairy Herds
Material type: Book ; Format:
Publisher: 2012 Dissertation note: Bovine viral diarrhea is one of the most important diseases of cattle which are causing
continuous economic losses to the cattle industry primarily due to decreased reproductive
I performance. Without doubt, direct contact between BVDV persistently infected, and susceptible animals is the most important transmission route of virus. All control programs which are in use in many countries of the world, mainly depend upon the detection of PI animals, eliminating them and preventing their return into the herds. Therefore, in this study diagnostic suitability of ear notch biopsies and serum samples were compared for the detection of PI animals, as well as proficiency of various diagnostic approaches like VI, AC-ELISA, IHC and real time RT-PCR were evaluated using ear notch biopsies. A total of 468 samples were collected from 12 participating dairy cattle farms located at Prince Edward Island, Canada. The samples were divided into two groups on the basis of age, A " 6 months), and B (> 6 months).
PI calves remain immunotolerant to the infecting strain but if exposed to a heterogonous strain postnatally, they may develop low level of antibody. Accordingly, serum neutralization was applied for initial screening of samples for further testing. The samples of animals of group B, having SNT (:S 1 :64) were selected, while all samples of younger aged group A were processed without considering the serum neutralizing titres, because unlike older animals, P.1. animals below 6 months of age can have passive colostral antibodies in the course of persistency. Diagnostic suitability of ear notch biopsy and serum sample for confirmation of BVDV
A significant discrepancy was observed between ear notch biopsies (51198 positive) and
serum samples (71198 positive) during first round of testing by real time RT-PCR. However, on
follow up testing, 30 days post first round of testing, a complete agreement between ear notch
biopsies and serum samples was observed. On second round of testing, a total of 4 animals out of
197 (one positive animals died before re-sampling) were confirmed with PI, using both ear notch
biopsies and serum samples. The decrease in the positivity using RT-PCR on serum samples in
the second round of testing reflected the presence of 2 transiently infected animals. Ear notch
biopsy (EN) testing did not detect any transiently infected animal indicating the lack of
delectability of the virus in EN during transient infection under conditions of this study. After
follow up testing, 2 animals in each of group A and B were identified as PI. These findings have
led us to conclude, that either serum or ear notch biopsy can be used for the detection of
persistent infection. Of 468 collected and 197 tested samples, an overall 0.85% and 2.03%
prevalence of PI animals with BVDV was observed respectively. A complete agreement (P value=l) was observed when three diagnostic approaches (Real time RT- PCR, AC-ELISA, and IHC) were compared with standard of VI. A total of 197 ear notch biopsies (145 of group A and 52 of group B) were tested by the four diagnostic tests, four animals (2 from group A and 2 from group B) were found positive by all the tests applied. A complete agreement was observed between the first and the second round of testing. All four assays were found specific but real time RT-PCR was found to be more sensitive. Both, VI and IHC were found labour intensive, as diagnosis may take more than one week to be made. Further PI calves remain immunotolerant tothe infecting strain but if exposed to a heterogonous strain postnatally, they may develop low leved
ofantibody. Accordingly, serum neutralization was applied for initial screening of samples for further testing. The samples of animals of group B, having SNT (:S 1 :64) were selected, while all samples of younger aged group A were processed without considering the serum neutralizing titres, because
unlike older animals, P.1. animals below 6 months of age can have passive colostral antibodies in the course of persistency. Diagnostic suitability of ear notch biopsy and serum sample for confirmation of BVDV persistent animals were evaluated by real time RT-PCR. TaqMan probes and primers specific for BVDVI and BVDV2 were used. They were found specific and able to detect 10·s and 10-4 TCID50 units ofBVDVI and BVDV2, respectively.
Availability: Items available for loan: UVAS Library [ Call number: 1407,T] (1).
Immunoprophylaxis Of Tick Infestation In Bovine
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Publisher: 2010 Dissertation note: A study to investigate prevalence of different genera of hard ticks was carried out in three districts of the Punjab province, Pakistan (Faisalabad, Jhang and Khanewal). Overall prevalence of Hyalomma species is the highest at 61 % as compared to other genera of hard ticks. In sex-wise distribution, it was found that female Hyalomma species were the highest at 85% followed by A mblyomma species at 81 %, whi Ie Rhipicephalus (Boophilus) species and Haemaphysalis species were at 77%. Infestation rate in cattle at 70% was higher as compared to buffaloes at 34%. In tick infestation level study, high infestation level in cattle at 59% was higher as compared to that of buffalo population at 18%. In cattle population, peR results showed the prevalence of T annulata in H anatolicum and Hidromedari ticks as 50% and 40% respectively. No theilerial organism was detected in Himarginatum, Rhipicephalus (Boophilus) annulatus and Amblyomma variegatum ticks. Three different types of vaccines were prepared from different organs of ticks i.e., salivary gland, intestine or whole ticks of the same species of Hyalomma and they were injected to rabbits. It was found that vaccine prepared by grinding whole tick produced the higher level of antibody as compared to two other vaccines. Each of the whole tick homogenate vaccine prepared from either of the species of Hyalomma, Rhipicephalus or Amblyomma and injected to rabbits. These vaccines produced antibody as well and cross reacted with each other showing each of the hard ticks were antigenically similar. Efforts were made to prepare oil based whole Hyalomma tick vaccine with three different antigen concentration 5.0 mg, 7.5mg and 10.0 mg and evaluated its potency in buffalo calves. It was found that the vaccine dose containing 5.0 mg antigen per dose did not produced detectable antibody in buffalo calves while the vaccine containing 7.5mg or more antigen produced detectable antibody. Moreover, we concluded that montanide based bard tick homogenate vaccine with more than 7.5mg protein per dose is effective in producing antibodies against tick infestation in the dairy animals. The antibody level in vaccinated buffaloes as well as invaccinated rabbits reached to peak level on day 45 post vaccination and started declining thereafter. Capacity of vaccine in controlling tick infestation was assessed in 12 cross-bred calves. It was found tbat rejection percentage in immunized group was higher as compared to control group. There was no difference of engorgement period between immunized and control group. Reproductive index in immunized group was lower as compared to control group. The efforts were made to grow midgut cells insect culture media after isolation them from Hyalomma Ticks.. The purpose of this experiment was to grow midgut cell and then use these cells as a source of was found that the vaccine dose containing 5.0 mg antigen per dose did not produced detectable antibody in buffalo calves while the vaccine containing 7.5mg or more antigen produced detectable antibody. Moreover, we concluded that montanide based bard tick homogenate vaccine with more than 7.5mg protein per dose is effective in producing antibodies against tick infestation in the dairy animals. The antibody level in vaccinated buffaloes as well as in vaccinated rabbits reached to peak level on day 45 post vaccination and started declining thereafter. Capacity of vaccine in controlling tick infestation was assessed in 12 cross-bred calves. It was found tbat rejection percentage in immunized group was higher as compared to control group. There was no difference of engorgement period between immunized and control group. Reproductive index in immunized group was lower as compared to control group. The efforts were made to grow midgut cells insect culture media after isolation them from Hyalomma Ticks.. The purpose of this experiment was to grow midgut cell and then use these cells as a source of contamination for the tick cell culture which are extrinsic as well extrinsic. The growth rate of these cells in our study was not optimal so the media was not splitted to get more cells.
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Identification Of Molecular Markers In Bmp15 Gene Of Different Pakistan Sheep And Goat Breeds
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; Nature of contents: Publisher: 2011 Dissertation note: Genetics of prolificacy in sheep and goat emphasize the importance of main genes which have been made known to affect litter size and rate of ovulation through various mechanisms. Natural mutations in prolific sheep and goat breeds have shown that the transforming growth factor beta (TGF-?) super family ligands such as bone morphogenetic protein 15 is crucial for ovulation and as well as for increasing litter size. Keeping in view the importance of prolificacy in sheep and goat, a research project was planed to identify the polymorphism, its association with fecundity and uncovering the nucleotide picture of BMP15 fecundity gene in sheep and goat breeds of Pakistan. In the research finding, various polymorphism, insertion and deletion of nucleotides in goat and sheep breeds of Pakistan were identified and associated with fecundity and secondly, some novel polymorphism in Pakistani goat and sheep breeds were identified which are different from the goat and sheep breeds of the world. This is the first report of the whole nucleotide of BMP15 gene in the sheep. A lot of work has been reported on these genes but total nucleotide picture in the sheep is not reported. Sequences of Bmp15 gene from sheep and goat breeds of Pakistan has been submitted to the NCBI GenBank database libraries,USA under accession numbers JN655669, JN655670, JN655671 and JN655672. It will result in fast vertical expansion of small ruminants to increase the mutton production and uplift the socio economic condition of small ruminant's farmers in the country.
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Epidemiology And Prophylaxis Of Babesiosis In Felidae
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Publisher: 2011 Dissertation note: Abstract
Availability: Items available for loan: UVAS Library [ Call number: 1425,T] (1).
Pharmacokinetics Of Ketoprofen In Domestic Animals
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Publisher: 2011 Dissertation note: Abstract
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Epidemiological, Haematological And Biochemical Risk Factors Of Parturient Haemoglobinuria In Buffaloes
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Publisher: 2011 Dissertation note: Parturient haemoglobinuria is disease of economic importance which affects a considerable number of buffaloes every year in India, Pakistan and Egypt. It is a non infectious hemolytic syndrome characterized by intravascular haemolysis, hypophosphataemia, haemoglobinaemia, haemoglobinuria and anaemia. The exact pathogenesis is not known and diversified etiological factors have been associated with this disease in different parts of the continent. Information on multidimensional etiological aspects of this buffalo syndrome is quite scanty. The present study was therefore carried out in district Chakwal for assessment of disease burden (parturient haemoglobinuria), its distribution and quantification of associated epidemiological, haematological and biochemical risk factors in order to suggest control measures and future research priorities.
Active surveillance was conducted in eight randomly selected villages of district Chakwal from April 2010 . March 2011. All breeding age buffaloes (1938) of these selected villages were taken as sampling frame whereas one breeding age buffalo was taken as sampling unit. Parturient haemoglobinuria appeared as number one disease among all problems of breeding age buffaloes with respect to mortality rate (1.03%) and proportional mortality rate (20%) whereas it appeared as 8th and 7th disease respectively with respect to incidence (3.97%) and case fatality (25.97%) rates.
Case-Control study was conducted for quantification of epidemiological risk factors associated with disease by analyzing the data of 180 case-control pairs for various 162
hypothesized risk factors. . 7 months pregnancy, . 3 lactation number, . 60 days postpartum period, . 7 years age, previous history of haemoglobinuria and ingestion of cruciferous plants were recorded as significant (P . 0.05) risk factors with odds ratios of 15.80, 6.39, 6.23, 5.56, 3.41 and 2.51 respectively. Clinical trial was conducted on 30 haemoglobinuric buffaloes randomly divided into three groups with 10 animals in each group to compare and assess the recovery rates of three different treatment packages against parturient haemoglobinuria. The highest recovery rate (100%) was recorded for combined therapy of sodium acid phosphate and blood transfusion followed by sodium acid phosphate with antifibrinolytic drug tranexamic acid (70%) and tranexamic acid with Novacoc injection (50%). Cross-sectional epidemiological study was conducted on haemoglobinuric (n = 30) and healthy (n = 60) buffaloes for quantification of haematological and biochemical risk factors associated with parturient haemoglobinuria. Red cell count (. 5 ~ 106 /ƒÊl), haemoglobin (. 8g / dl), haematocrit . 25%, mean corpuscular volume (. 50fL), mean corpuscular haemoglobin (. 20pg) and erythrocyte sedimentation rate( . 80mm / 1st hour) were recorded as significant (P . 0.05) haematological risk factors with odds ratios of 26, 17.81, 28.95, 21, 12.25 and 26 respectively whereas billirubin unconjugated (. 0.2mg /dl), billirubin total ( . 0.3mg /dl), phosphorous (. 2.5mg /dl), molybdenum (. 70ƒÊg /dl) and selenium (. 15 ƒÊg /dl) were recorded as significant (P.0.05) biochemical risk factors with odds ratios of 26.55, 26.55, , 7.50, 11 respectively.
Experimental study was conducted to determine the effect of orally administered gossypol on haematological and biochemical parameters of eight female rabbits of six 163
months age purchased from local market and maintained at university of veterinary and animal sciences from February 2011 . April 2011 under optimum conditions. The cotton seed cake containing free gossypol contents of 0.25% was fed to rabbits @ 4 grams per kg per day in addition to their routine diet including good quality fresh vegetables (cucumbers, spinach, cabbage & carrots) and clean water ad-libitum. Blood samples of each rabbit were collected after every 15 days interval and analyzed for haematological and serum biochemical parameters. Significant (P.0.05) decrease was recorded in total erythrocyte count, haemoglobin concentration, haematocrit, mean corpuscular volume and serum inorganic phosphorous whereas significant increase was recorded in mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, red cell distribution width, total leukocyte count, lymphocytes and monocytes from 0 . 60th day with the passage of time whereas non significant (P.0.05) difference was recorded with respect to granulocytes and serum calcium concentration.
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Effect Of Protein Supplements Of Varying Ruminal Degradability On Milk Production, Composition And Nutrients
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Publisher: 2012 Dissertation note: The study on "Effect of protein supplements of varying ruminal degradability on milk production, composition and nutrients utilization in early lactating Sahiwal cows and Nili-Ravi buffaloes" was carried out in three phases at three different experimental locations. The in situ study of animal and vegetable protein sources was conducted at the Department of Food and Nutrition, University of Veterinary and Animals Sciences, Lahore while the feeding trials with early lactating Sahiwal cows and Nili-Ravi buffaloes were carried out Government Livestock Farm, Kalurkot, Bukkar and Livestock Experimental Station, Khushab, respectively.
Different animal (n = 6) and vegetable origin (n = 15) protein sources were subjected to ruminal protein degradability analyses using the in situ technique. All these test feeds collected from ten different locations were subjected to ruminal incubation (in triplicate) for 0, 3, 6, 12, 24 and 48 h to determine the quickly soluble fraction (a), potentially degradable fraction (b), degradation rate (c) and effective degradability at different (2, 5, 8 %) ruminal passage rates. The degradability characteristics in animal protein sources (part 1, phase 1) showed significant differences in degradation kinetics and effective degradability (ED). In crude protein (CP) degradability, the quickly soluble fraction (a) was higher (P<0.05) in fish meal, PBM and meat meal and lower (P<0.05) in blood meal, feather meal and bone and meat meal. Potentially degradable fraction (b) among test feeds was maximum (P<0.05) in bone and meat meal and PBM and minimum (P<0.05) in blood meal and feather meal. The degradation rate (c) did not differ among the test feeds. Of all the animal protein sources investigated, meat meal showed maximum CP degradability at 0.05 rumen passage rate whilst, minimum (P<0.05) ED of CP was exhibited by blood meal.
Ruminal degradability characteristics in vegetable protein sources (part-2 of phase-1) showed variation in degradation kinetics and ED of CP. The quickly soluble fraction (a) was highest (P<0.05) in sesame cake and lowest (P<0.05) in CGM 60%, coconut meal and PKC. Potentially degradable fraction (b) was maximum (P<0.05) in CGM 60%, PKC, SBM and guar meal while minimum (P<0.05) in sesame cake and CGM 30%. Protein degradation rate (c) was highest (P<0.05) in CSC while lowest (P<0.05) in coconut meal, coconut cake and CGM 60%. Effective degradability of CP at 0.05 rumen passage rate was highest in sesame cake and lowest (P<0.05) in coconut meal.
All vegetable protein sources were treated (part-3 of phase-1) with formaldehyde (1 g/100 g CP) and heat treatment (1 h at 15 lb/100 g CP) to determine their effectiveness in reducing ruminal protein degradability. Both of these treatments decreased (P<0.05) rumen degradability of the vegetable protein sources investigated. Of the formaldehyde treated test feeds, quickly soluble fraction (a) was higher (P<0.05) in sesame cake and lower (P<0.05) in CGM 60%, SBM, CGM 30%, guar meal, canola meal and coconut meal. The highest value of potentially degradable fraction (b) was recorded (P<0.05) in CSC and RSC while CGM 60% had the lowest value (P<0.05). Degradation rate (c) was highest (P<0.05) in RSM, RSC, CSC, CSM coconut cake, PKC, sesame cake, SFM and CGM 60% and lowest (P<0.05) in CGM 30%, guar meal and canola meal. Effective degradability of CP was maximum in sesame cake at all the rumen passage rates. In contrast, CGM 60% had the lowest (P<0.05) ED at all of the rumen passage rates. Among the heat treated vegetable protein sources, quickly soluble fraction (a) was highest (P<0.05) in sesame cake and lowest (P<0.05) in CGM 60% and SBM. Potentially degradable fraction (b) had the highest (P<0.05) value in almond cake, RSM, RSC, CSC and SFM while CGM 60% had the lowest value (P<0.05). Effective CP degradability of the heat treated test feeds showed that almond cake and sesame cake had the highest (P<0.05) ED whilst CGM 60% had the lowest values (P<0.05). In comparing both treatments, similar influence (P>0.05) of increasing RUP level was recorded in CGM 30%, SFM, RSM, CSM, PKC and coconut meal. Formaldehyde treatment was found more effective (P<0.05) in increasing RUP level in guar meal, canola meal, RSC, CSC, coconut cake, almond cake and sesame cake whilst heat treatment increased (P<0.05) RUP level in SBM and CGM 60% at applied rates in this study.
In phase-2, a feeding trial with early lactating Sahiwal cows was conducted to investigate the effect of protein supplements of varying ruminal degradability on milk production, composition and nutrients utilization. Twenty four early lactating Sahiwal cows were selected and randomly divided into four groups. Four iso- caloric and iso- nitrogenous diets i.e., ration A (30% RUP), ration B (40% RUP), ration C (50% RUP) and ration D (60% RUP) were fed in a completely randomized design. Dry matter and CP intakes were significantly affected by ration composition (P<0.01), whereas NDF and ADF intakes did not vary among the four treatment groups (P>0.05). DM intake was higher (P<0.05) in cows receiving rations B and A than the cows fed rations C and D. There were significant differences in DM (P<0.05), CP (P<0.001) and NDF (P<0.05) digestibility due to the ration; however, ADF digestibility did not differ (P>0.05) between the rations. DM digestibility was higher (P<0.05) on ration B than rations C and D, but similar to that for ration A. Crude protein was higher (P<0.05) on rations B and A and lower (P<0.05) on rations C and D. Daily yields of uncorrected milk and protein were highest in early lactating Sahiwal cows fed ration B and lowest when fed ration D. Daily yields of 4% FCM and milk fat were higher (P<0.05) on rations B and A and lower (P<0.05) on ration D. In milk composition, fat, protein and total solids contents were the same across all diets. Nitrogen intake was highest (P<0.01) for rations B and A and lowest for ration D and C. Nitrogen balance (g/d) and as a percentage of N intake varied; with the cows consuming ration B retaining maximum (P<0.001) N. However, N balance did not vary between rations A, C and D. Nitrogen utilization was highest (P<0.001) in cows fed ration B, but there was no difference among cows fed rations A, C and D. Live weight and body condition score in cows were unaffected by the rations. Cost of milk production was least on ration B and highest on ration D.
In phase-3 a feeding trial using early lactating Nili-Ravi buffaloes was conducted. Twenty four buffaloes were selected and randomly divided into four groups. These groups were fed four experimental diets i.e., rations A, B, C and D having 30, 40, 50 and 60% RUP proportions, respectively in a completely randomized design. Results showed no differences (P>0.05) in the intakes of DM, CP, NDF and ADF intake between the rations. Likewise, DM, CP and ADF digestibility were the same (P>0.05) in buffaloes fed rations A, B, C and D; however, NDF digestibility was higher (P<0.01) on ration C and B while lowest on rations A and D. Milk yield was highest (P<0.001) on ration C while lowest (P<0.001) on rations D and A. Buffaloes given ration C produced more (P<0.05) FCM than those receiving rations A, B and D. Daily yield of milk fat was greater (P<0.001) on ration C compared to the other three rations. Milk protein yield was highest (P<0.001) on ration C and lowest (P<0.001) on rations A and C. Diet had no effect (P>0.05) on milk fat, SNF, lactose, salts and total solids percentages; whilst milk protein percentage varied among all four diets, viz ration C>ration B>ration D>ration A. Nitrogen, intake, nitrogen balance and nitrogen utilization were similar across all the diets. Live weight and body condition score in buffaloes were unaffected by the diet fed. The cost of milk production was highest (P<0.05) with rations D and B whilst lowest (P<0.05) on ration C.
It is concluded that among animal protein sources rumen CP degradability was least in blood meal and maximum in meat meal. In vegetable protein sources, coconut meal showed least ruminal CP degradability while sesame cake recorded with highest ruminal CP degradability. Both formaldehyde and heat treatments protected protein from ruminal degradability with varied levels of effectiveness in different feeds. Production performance improved with the use of RUP sources in early lactating cows and buffaloes. Sahiwal cows showed better yield performance in diets having 40% un-degradable protein in the diet, while Nili-Ravi buffaloes showed high yield performance in diets with 50% un-degradable protein sources.
The use of latest technology and methods needs to be applied for minimizing variations involved in evaluating CP degradability of feeds through in situ procedure. Influence of RDP and RUP based rations in mid and late lactation of Sahiwal cows and Nili-Ravi buffaloes are also fertile areas of research. The studies on degradability of amino acids for compiling 'internal standards' of feed resources and production performance of lactating cows/buffaloes based on ruminal degradability of amino acids rather than protein degradability would be better approach for future studies.
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Feeding Behavior And Performance Of Sheep And Goats Under Various Feeding Management Systems
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Publisher: 2012 Dissertation note: Sheep and goats have been bestowed with the capacity of surviving under a variety of environmental conditions including the coastal region, plains and high mountains. The profitable small ruminants farming depend upon feeding and management systems because the feed cost is 70% in any livestock farming. In Pakistan, mostly people grazed ruminants on summer and winter fodders for maintenance and production requirements. Commercial livestock production demands a change in feeding with a trend for more efficient utilization of scarce feed resources. The proposed study was planned in to three experiments under a factorial arrangement to evaluate the growth performance of sheep and goats. In experiment one a study was first conducted to compare the voluntary intake and digestibility of janter (coriandrum sativum), guar (cyamopsis tetragonolba), cowpea (Vigna sinesis) in sheep and goats. For this purpose, 90 female animals (sheep n=45 and goats n = 45) were selected randomly and divided equally in, 6 groups representing each species under 2×3 factorial arrangements, Groups A,B ,C represented goats while group D,E,F represented sheep. Results showed that goats spent more time on eating than sheep while ruminating time was higher in sheep than goats. Drinking time was not different (P>0.05) among the species. Goats spent more time on playing and resting than sheep fed guar, cowpeas and jantar. Dry mater CP, NDF, ADF and GE intake was higher in sheep than goats fed guar, cowpeas and jantar. DMD and CP were higher in sheep than goats fed guar. NDF and ADF digestibility was similar in both species. Average daily weight gain, feed efficiency and cost of gain were similar in both the species. It is concluded that the jantar fodder in summer is most suitable fodder for sheep and goats compare to guar and Cowpea. In the second trial of the first phase study comparison of voluntary intake and digestibility maize (Zea mays), sorghum (Sorghum bicolor) and millet (Pennisetum americanum) in sheep and goats were compared. Statistical analyses showed that eating time was higher (P?0.05) in goats than sheep fed maize, millet and sorghum while, sheep spent more time on ruminating, drinking and standing than goats. Goats showed higher playing, resting and other activities than sheep fed maize, millet and sorghum. Dry matter CP, NDF and ADF intake was similar (P>0.05) in both the species fed maize, millet and sorghum. Dry matter digestibility was similar in sheep and goats fed maize, millet and sorghum. NDF digestibility was similar (P>0.05) in goat and sheep fed sorghum while this was different (P?0.05) when maize and millet were fed. ADF digestibility was similar (P>0.05) in goat and sheep. Average daily weight gain feed efficiency and cost of gain was not significant (P>0.05) among both the species fed maize, millet and sorghum. Results of the study showed that the non leguminous fodders during summer are equally preferred by both species. In second the phase voluntary feed intake and digestibility of berseem, (Trifolium alexandrium) lucerne, (Medicago Sativa), oats, (Avena Sativa) in female sheep and goats was studied. For this purpose, female animals (n=90) of sheep (n=45) and goats (n=45) were randomly selected and divided equally in six in a 2×3 factorial arrangement. Results showed that eating time was higher (P<0.05) in goats than sheep, while ruminating time was more in sheep than goats fed berseem lucerne and oats, whereas time spent on drinking was similar in both goats and sheep. Goats utilized less time in standing, higher (P<0.05) time in playing, resting and other activities than sheep fed maize, millet and sorghum. Crude protein intake was higher (P<0.05) in goats than sheep fed berseem and lucerne. DM intake was higher (P<0.05) in goats than in sheep fed berseem, while it was similar when fed lucerne and oats fodder. NDF, ADF and GE (M cal/d) intakes were higher (P<0.05) in goats than sheep fed berseem and lucerne fodder however it was similar in both the species fed on oats fodder. DM digestibility was similar (P>0.05) in sheep and goats fed berseem, lucerne and oats. CP digestibility was higher (P<0.05) in goats than in sheep fed berseem. When fed Lucerne and oats there was no significant difference (P>0.05) between goats and sheep.. NDF digestibility was higher (P<0.05) in goats than in sheep fed berseem. Average daily gain, feed efficiency and cost of gain/kg was non-significant (P>0.05) between goats and sheep fed berseem, lucern and oats. Results demonstrated that during winter the most suitable fodder for sheep and goats is lucerne fodder. In the second experiment the study was conducted to compare the performance of sheep and goats under various feeding management systems in which ninety female animals were selected and divided into six equal groups with three groups of each species (sheep n=45, goats n=45) under a 2×3 factorial arrangement. These were in extensive, semi-intensive and intensive feeding management systems. Dry matter intake was higher (P?0.05) (P<0.05) in sheep than goats kept under extensive, semi-intensive and intensive systems. Crude protein intake was significantly higher (P<0.05) in sheep than goats fed intensively. NDF and ADF intake was higher (P?0.05) (P<0.05) in sheep than in goats. Average daily weight gain was higher in sheep than goats on the extensive system followed by the semi-intensive system. Feed efficiency was similar in goats and sheep while the cost of gain per kg was more economical in sheep than goats. Results of study revealed that both species performed better on extensive feeding system than the other systems might be of natural grazing behavior. The third experiment of study was conducted to compare the performance of sheep and goats under the intensive management system. Sixty female animals (lambs n= 30 and kids n=30) were used. The animals were divided equally in four groups A and B representing lambs while C, D was for kids. Both species were allotted two treatments i.e. fodder ad libitum with concentrate supplement (240 grams/animal/day) and total mixed ration ad libitum under a 2×2 factorial arrangement. Results showed that DM, CP, NDF and ADF intakes were higher (P?0.05) in lambs than kids. Average daily weight gain was higher (P?0.05) in lambs than kids fed total mixed ration. Feed efficiency was higher (P?0.05) in kids than in lambs fed fodder plus supplement. Dry matter and CP digestibility was higher (P?0.05) in kids than lambs fed a total mixed ration. NDF digestibility was maximum (P?0.05) in lambs than kids fed the TMR, it was also higher in kids than in lambs when fed fodder plus the concentrate supplement. ADF digestibility was maximum (P?0.05) in lambs than in kids fed the total mixed ration. The performance of lambs was better on TMR while kids showed good results on fodder plus the concentrate supplementation.
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Studies On Genetic Diversity Of Labeo Rohita And Cirrhinus Mrigala By Using Molecular Markers In Punjab-Pakistan
Material type: Book ; Format:
Publisher: 2012 Dissertation note: The studies on genetic diversity of Labeo rohita and Cirrhinus mrigala by using molecular markers in Punjab-Pakistan were carried out to investigate the genetic structure of said Indian major carps by RAPAD marker and the levels of polymorphism and similarity amongst the different groups of five populations of wild and farmed types. The results obtained from the present study after statistical analyses are presented in section-4 of this dissertation. The samples were collected from the following sites; for farmed fish was collected from UVAS-Fish Hatchery, C-block Ravi campus Pattoki district Kasur and for wild fish; from Trimu Barrage at the junction of River Chenab and Jhelum near district Jhang, Taunsa Barrage at River Indus near tehsil Kot Adu district Muzaffar Garh, Qadirabad Barrage at River Chenab near district Mandi Bahuddin and Baloki Barrage at River Ravi near tehsil Bhai Phero district Kasur. Analysis of variance (ANOVA) for the different morphometric parameters of study and Pearson's correlation among the physico-chemical parameters of water quality was done by Minitab statistical computer software. The XLSTAT 2012 version 1.02 of the computer software was used for the Pearson correlation analysis of the morphometric parameters of study. The same computer program was used for Agglomerative Hierarchical Clustering (AHC) of the different genotype occurrence on the basis of differences in morphometric parameters was done by Agglomeration method by following the Unweighted Pair Group Method with Arithmetic Mean (UPGMA). The Principle Component Analysis (PCA) on the basis of differentiation in morphometric parameters by Eigenvalues and differentiation into factors of the different genotypes from the different environmental conditions was done by correlation bi-plot/coefficient of the correlation (n) method in the same program. This software was also used to analyze the RAPAD data for Jaccard's coefficient by following the Unweighted Pair Group Method with Arithmetic Mean (UPGMA) for Hierarchical Clustering of the similar groups on the basis of similarity amongst the genotypes and the dendrogram generated is presented in the next section. The Principal Component Analysis (PCA) for grouping of the different genotypes from the different environmental conditions was done by Spearman Varimax rotation method for bi-plot generation of the co-occurrence of the same genotypes with similar genetic properties and specificity of different primersin the same program.
Following results were obtained:
1. Morphometric parameters of L. rohitashowed following trends: body weight, total length and average length of paired pectoral fins were nonsignificantly different (p > 0.05), fork length, dorsal fin length, caudal fin length and average length of paired pectoral fins were highly significantly different (p < 0.01) while anal fin length was significant different (p < 0.05) among the experimental sites. In case of C. mrigala, the body weight was non-significantly different (p > 0.05) while all other parameters were highly significantly different (p < 0.01) except the dorsal fin length which was significantly different (p < 0.05) among the study sites.
2. The results of the Pearson correlation of morphometric parameters showed that body weight of L. rohita developed a positive and highly significant (p< 0.0001) correlation with all the remaining morphometric parameters, the fork length of the said species showed a positive and highly significant (p< 0.0001) correlation with all the parameters except with the caudal fin length where the correlation was also positive but non-significant (p = 161). In case of total length of the fish body, the correlation was highly significant (p< 0.0001) and positive with all the parameters of study.The length of the dorsal fin showed highly significant (p< 0.0001) and positive correlation with all the remaining morphometric parameters under study. The caudal fin length of L. rohita showed a positive and highly significant (p< 0.0001) correlation with all the other parameters except the fork length where the correlation was positive but non-significant (p = 161). The correlation of the anal fin length of the fish body showed a highly significant (p< 0.0001) and positive correlation trends. The average length of the paired pectoral fins showed a positive and highly significant (p< 0.0001) correlation with all the remaining morphometric parameters of study, the correlation of paired pelvic fins average length showed positive and highly significant (p< 0.0001) correlation with other parameters.
3. The body weight of C. mrigala developed a positive and highly significant (p< 0.0001) correlation with all the remaining morphometric parameters. The fork length of the said species showed a positive and highly significant (p< 0.0001) correlation with body weight, total length and dorsal fin length while this correlation was positive but non significant with the caudal fin length (p = 0.228), anal fin length (p = 0.168), average length of paired pectoral fins (p = 0.031) and average length of the paired pelvic fins (p = 0.106). In case of total length of the fish body, the correlation was highly significant (p< 0.0001) and positive with all the parameters of study. The length of the dorsal fin showed highly significant (p< 0.0001) and positive correlation with all the remaining morphometric parameters under study. The caudal fin length of C. mrigala showed a positive and highly significant (p< 0.0001) correlation with all the other parameters except the fork length where the correlation was positive but non-significant (p = 0.228).The correlation of the anal fin length of the fish body showed a highly significant (p< 0.0001) and positive correlation trends with all the parameters except the fork length where the correlation was positive but non-significant (p = 0.168). The average length of the paired pectoral fins showed a positive and highly significant (p< 0.0001) correlation with all the remaining morphometric parameters of study except the fork length where the correlation was positive but non-significant (p = 0.031). InC. mrigala, the correlation of paired pelvic fins average length showed positive and highly significant (p< 0.0001) correlation with other parameters except the fork length where the correlation was positive but non-significant (p = 0.106).
4. Dendrogram generated on the basis of morphometric parameters of study dividedL. rohita genotype in to five major clusters or classes with 19.24% for within class variation while 80.76% for the between class differences. While the dendrogram developed for C. mrigala divided the genotypes in to four major clusters or classes with 27.28% for within class variation while 72.72% for the between class differences.
5. The results obtained from the PCA for morphometric parameters of L. rohitaand C. mrigalaindicated clearly that the increase in the number of factors or components was correlated with the decrease in eigenvalues. The values showed that its trend reached its maximum at level of second factor. In the same way according to the Kaiser (1958) criterion based upon the eigenvalues greater than one, first two main factors accounted for 80.273% of cumulative variability for L. rohita and 82.558% for C.mrigala. The PCA grouped the tested variables or parameters of the L. rohita,the first group amongst the major two groups accounted for 64.245% of the cumulative variability while the second from these accounted for 16.028% of the cumulative variability. The PCA grouped C. mrigala,also into two groups, the first group amongst the major two groups accounted for 59.323% of the cumulative variability while the second from these accounted for 23.235% of the cumulative variability.
6. The physico-chemical parameters of the water samples of all study sites were analyzed for correlation among them. The results were as follows; the correlation of the pH with water temperature (r= 0.107) and dissolved oxygen (r = 0.905) was positively non-significant while the correlation with electrical conductivity (r = -0.798), salinity (r= -0.888), total dissolved solids (r = -0.857), total alkalinity (r = -0.736) and total hardness (r = -0.499) was negatively non-significant. The correlation of the dissolved oxygen with water temperature (r= 0.313) was positively non-significant while the correlation with electrical conductivity (r = -0.669), salinity (r= -0.828), total dissolved solids (r = -0.809), total alkalinity (r = -0.930) and total hardness (r = -0.300) was negative but also non-significant as like with the water temperature. The electrical conductivity was positively correlated with all the physic-chemical parameters as with water temperature (r= 0.482), salinity (r= 0.925), total dissolved solids (r = 0.889), total alkalinity (r = 0.452) and total hardness (r = 0.906) and this correlation was non significant.The salinity amongst the water parameters was correlated positively with water temperature (r = 193), total alkalinity (r = 0.717) and total hardness (r = 0.734) and it was non-significant but with total dissolved solids (r = 0.994) the correlation was also positive but highly significant (P < 0.001). The total dissolved solids values observed from the study sites were positively correlated with water temperature (r = 0.172), total alkalinity (r = 0.734) and total harness (r = 0.657) and this correlation was non-significant. The correlation between the total alkalinity and total hardness was also positive and non-significant (r = 0.048).
1. In case of L. rohita, OPB-1 polymorphism remained as 16.67%, OPB-3 polymorphism remained as 40.00%, OPB-4, polymorphism remained as 16.67%, OPB-5 polymorphism remained as 20.00%, OPB-7 polymorphism was 28.57%, OPB-8 polymorphism was 20.00%, OPB-9 polymorphism was 25.00%, OPB-10 polymorphism was 28.57%, OPC-19 polymorphism was 14.29% and OPD-4 showed 50.00% polymorphism in amplification. In case of C. mrigala, OPB-1 polymorphism remained as 16.67%, OPB-3 polymorphism remained as 16.67%, OPB-4 polymorphism remained as 25.00%, OPB-5 polymorphism remained as 14.29%, OPB-7 polymorphism was 14.29%, OPB-8 polymorphism was 20.00%, OPB-9 polymorphism was 20.00%, OPB-10 polymorphism was 20.00%, OPC-19 polymorphism was 28.57% and OPD-4 polymorphism remained as 33.33% in amplification.
2. The dendrogram generated by UPGMA of RAPAD data of L. rohita by the randomly selected individuals with high scorable bands of the five populations grouped themselves in the first class/cluster while a single sample designated as Indus2 from the population from River Indus collected from Taunsa Barrage represents the second class/cluster and in same way only single individual designated as Ravi2 collected from River Ravi from the Baloki Barrage represents the third class. The dendrogram generated by UPGMA of RAPAD data of C. mrigala by the randomly selected individuals of the five populations grouped themselves in the first class/cluster and two samples designated as Indus2 and Qad2 from the populations from River Indus collected from Taunsa Barrage and River Chenab from Qadirabad Barrage represents the second class/cluster while one individual from the Trimu Barrage at the junction of Jhelum and Chenab Rivers designated as Trimu2 represents the third class and in the same way only single individual designated as Ravi2 collected from River Ravi from the Baloki Barrage represents the third class.
3. The PCA resultsfor L. rohitait can be assumedthat PCA grouped the tested variables or parameters of the fish RAPAD amplification data into two main components which all together accounted for 58.177% of the cumulative variation among the factors. The first group (F1) amongst the major two groups accounted for 33.327% of the cumulative variability while the second (F2) from these accounted for 24.850% of the cumulative variability. These results were also confirmed after the varimax rotation. By the PCA resultsfor C. mrigalawe can assume after observing the results that the PCA grouped the tested variables or parameters of the fish RAPAD amplification data into two main components which all together accounted for 70.866% of the cumulative variation among the factors. The first group (F1) amongst the major two groups accounted for 51.115% of the cumulative variability while the second (F2) from these accounted for 19.751% of the cumulative variability.
This study in this way has provided the genetic information of the present fish species and how evolutionary processes are affecting the fish fauna. So this study along with the strengthening of the academic research area has also proven an applied research which will help the breeders to the chose most fit candidates for the breeding program in the Pakistan.
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Epidemiology, Zoonotic Potential, Haematiology And Therapy Of Toxocariasi In Dogs And Humans.
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Publisher: 2010 Dissertation note: Abstract
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Chemical, Microbiological And Toxicological Screening Of Tannery Effluent Wastewater
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Publisher: 2012 Dissertation note: Over the last decade or so the chromium based tanning industry has shown rapid growth in Pakistan. However the rule and regulations promulgated by the government are not strictly followed for the processing of effluent discharged by the tanneries. Consequently tannery effluents have become a great source of water pollution in surrounding area. This project was designed to evaluate the hazardous effects of tannery effluent wastewater (TEW) through various bioassays.
During the first phase of the project, composition of the TEW samples was determined by PIXE analysis. Besides this, we have also investigated the impact of TEW on trace element content of ground water in Kasur tannery area. The ground water from shallow tubewells (100 to 300 ft) in the area has shown very high content of chromium while the ground water from the deeper tubewells (upto 600 ft) generally does not contain the toxic elements except for one outlet of the water supplied by the Muncipal Corporation. This could be due to corroded pipes in the tannery area.
Microbial load was determined during second phase of this research project by viable count method. The detected viable count was 7.5 X 104 to 3.0 X 107CFU/ml. Various strains of chromium tolerant bacilli were isolated and they were found tolerant up to 2600 µg/ml supplemented chromium sulphate.
During the third phase of this research plan, dilutions of TEW were evaluated for their effects on angiogenesis using CAM assay. TEWD1 and potassium dichromate were found highly anti-angiogenic. Moreover, dilutions of TEW and potassium dichromate have demonstrated significant toxicity when assessed through marine shrimps mortality assay and phytotoxiciy assasy.
Chronic toxicity study on Wistar rats was conducted in the last phase. Chronic exposure of TEW for three months to rats leads to the development of various lesions in lung, liver, kidney and heart of rats.
In short, TEW and contaminated ground water of Kasur is imposing a great threat not only to local inhabitants of the city but also to the population of far distance.
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Molecular And Serological Characterization Of Avian Influenza Viruses In Domestic And Wild Birds
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Publisher: 2012 Dissertation note: Avian Influenza virus (AIV) has been recognized one of the most important pathogen in poultry industry. AIVs play an important role in the pandemic spread that could cause high morbidity and mortality in human beings. A total of 1500 tracheal and cloacael swabs were collected from the seven live bird poultry markets of Lahore Pakistan for surveillance of AIV. The samples were processed for virus isolation in chicken embryos. The isolates were processed for HA test and AIV Antigen Rapid Test Kit to differentiate Newcastle disease virus and Avian influenza virus. Only four samples were positive for Avian influenza H9N2 subtype and 17 were positive for Newcastle disease virus. Four HA virus suspension of AIV showed high titers of anti AIV H9N2-HI antibody titer in chicken as well as rabbit serum, raised using Ottoman Pharma AIV-H9N2 (Oil based) vaccine.
The isolates of AIV H9N2 were confirmed using laboratory optimized RT-PCR, mRT-PCR and LAMP tests. All isolates were sequenced and analyzed to develop phylogenetic relationship. Two isolates A/Chicken/Pakistan/Micro-1/2009 and A/ chicken/ Pakistan/ Micro-2/ 2009 showed 99.1% nucleotide homology with each other and 95- 99% homology with the other Pakistani isolates, 95.1% homology with A/Chicken/Iran/B102/2005. The nucleotide sequence of "A/Duck/Pakistan/Micro-3/2009" showed 98.8% homology with "A/chicken/Pakistan/micro-4/2009", 98-98.7% homology with other Pakistani Isolates and 95.8% homology with "A/Chicken/Iran/B102/2005". The nucleotide sequence of "A/Chicken/Pakistan/Micro-4/2009" showed 99.6% homology with "A/duck/Pakistan/micro-3/2009", 95-96% homology with other Pakistani isolates and 94.3% homology with "A/Chicken/Iran/TH lBM863/2007".
Three out of four isolates had PARSSRGL cleavage sites and one isolate A/chicken/Pakistan/micro-1/2009 had PAKSSRGL cleavage site. The four isolates of the study contained a 226-Leu and 228-Gly at receptor binding sites. Substitution of Glutamine (Q) into Leucine (L) at 226 receptor binding site in HA glycoprotein increased the binding specificity for Sialic acid ? 2, 6 Galactose linkage of human receptor. The HA gene of the live poultry market isolates had 7 predicted glycosylation sites at 29-32, 105-108, 141-144, 298-301, 305-308, 492-495 and 551-554, positions. The NA gene of the live poultry market isolates had 8 predicted glycosylation sites at 44-46, 61-63, 69-71, 146-148, 200-203, 234-237 and 402-403, positions. Predicted glycosylation sites affected the structure and stability of NA protein.
It is concluded that continuous epidemiological and virological surveillance of live bird poultry markets may help scientists to develop an effective control and preventive measures for AI viruses.
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Epidemmiology Of Foot And Mouth Disease In Buffaloes Of Punjab Province
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Publisher: 2009 Dissertation note: This study indicates that the ranking order of buffalo diseases, with respect to their incidence in descending order in Punjab province is Foot and Mouth Disease, Mastitis, Diarrhea, Haemorrhagic Septicemia, Sudden Death Syndrome and haemoglobinuria. Similarly the disease ranking order in cattle in descending order is FMD, Mastitis, Diarrhea, Hemorrhagic Septicemia, Haemoglobinuria and Sudden Death Syndrome. FMD is top most economic important disease both in buffaloes and in cattle in the province.
Morbidity rate in the adult cattle and buffalo was higher as compared to the younger stock. However, the mortality rate was higher in young stock as compared to the adult animals of both the species. Moreover, adult and young males of both the species were more susceptible to the disease as compared to females.
Cross-sectional survey revealed the economic loss of Rs. 41.32 million due to loss of milk, cost of dead animals and treatment cost of sick and complicated cases of FMD. The loss due to milk reduction was 57.3% of the total losses followed by mortality loss (26.4%), morbidity effect expenses (15.2%) and treatment charges in FMD complicated cases (1.0%). The findings of present study clearly indicate the association of age, feeding pattern, vaccination status and season as risk factors in the incidence of FMD in Punjab.
Data obtained from the EPI-Unit Lahore showed that 719 FMD outbreaks occurred in the district of Punjab during 2007-2008. The highest number of outbreaks (212) was recorded in Rahim-Yar-Khan followed by Bhakkar (118), T.T. Singh (81) and Faisalabad (72). Of the total 309 disease outbreaks in buffalo, 174 (56.3%) were recorded in adults, whereas this number in cattle was 169 (61%). The incidences of the outbreaks increased gradually following the post-monsoon period. The greatest number of outbreaks was observed during the winter season, from December to February. Data from FMD Research Center, Lahore revealed the involvement of only FMDV serotype "O" in all the outbreaks during 2007-2008.
Studies of the factors (age, feeding pattern, stage of pregnancy and species) on the immune response of local trivalent FMD vaccine revealed that buffaloes of all age groups responded well to vaccination against disease. It was also observed that 7-9 months pregnant buffaloes elicited significantly lower antibody response to vaccine as compared to the control groups. Similarly, buffaloes on grazing have shown lower anti-FMD-CF GM titer as compared to buffaloes on manger feeding. Sheep and goat were found to be late and poor responder to vaccine as compared to cattle and buffalo.
Analysis of 300 serum samples from FMD affected buffaloes of 12 districts of the Punjab indicated the highest incidence of serotype "O" (62.3%) followed by Asia-1 (32.4%) and "A" (3.30%) in the population tested.
FMD virus was inactivated at 61 ºC within 15 minutes and at pH 4, 8, and 10 within 24 hours. However, ultraviolet radiation was unable to inactivate the virus even after 45 minutes. The disinfectants/chemicals evaluated in this study including sodium hydroxide, sodium carbonate, citric acid, acetic acid, formalin, sodium hypochlorite, virkon-s, aldekol and Gas-G were effective in inactivating the FMDV at recommended concentration levels of 2%, 4%, 0.20%, 4%, 0.15%, 3.0%, 1.0%, 0.50% and 0.1% after 60, 30, 60, 60, 30, 30, 30, 60 and 30 minutes, respectively, at 300C. Sodium hypochlorite and Gas-G were equally good in inactivating the virus at half (1.5% and 0.05%) of the recommended concentration.
Efficacy trial of local and imported oil based trivalent FMD vaccine in six villages, of the Faisalabad district clearly showed that 81.8% of FMD cases were prevented by the local inactivated vaccine in vaccinated animals whereas; this percentage was 70.6 in case where imported vaccines were employed. Moreover, efficacy of the local vaccine was higher than the imported vaccines.
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Evaluation Of Different Extenders For The Cryopreservation Of Buffalo Bull (Bubalus Bubalis) Semen
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Publisher: 2011 Dissertation note: Buffalo is playing an important role in our country's economy by producing milk, meat and draught power. Genetic potential of low producing animals can be improved by using artificial insemination technology. Unfortunately, less number of elite bulls are available and low fertility rate of buffalo by using cryopreserved semen has been obtained. Semen is exposed to osmotic and oxidative stresses during processing, cryopreservation and thawing before insemination. Fertilizing ability is lost due to spermatozoa damage and it ultimately results in poor conception rates in buffalo. In order to protect spermatozoa from these stresses and improve fertility in buffalo, five osmotic pressure based concentrations of three extenders i.e. Citrate egg yolk extender (CEYE), Tris egg yolk extender (TEYE), and Lactose egg yolk extender (LEYE) were prepared by varying the quantity of the solutes to obtain an osmotic pressure of 255, 265, 275, 285 and 295 mOsm/kg. Osmotic pressure was measured by an osmometer.
In the first experiment, equal volume of semen obtained from four Nili-Ravi buffalo bulls was pooled and used to study the effects of osmotic pressure on post thawed semen characteristics. For this purpose, three basic media: citrate fructose media, tris citric acid fructose media and lactose media were prepared and divided each media in to five equal parts to maintain osmotic pressures of 255, 265, 275, 285 and 295mOsm/kg. These basis media were stored in a biomedical freezer, which were later used in preparing three semen extenders i.e. Citrate egg yolk extender (CEYE), Tris egg yolk extender (TEYE), and Lactose egg yolk extender (LEYE). During each collection, fifteen extenders (each of three extenders having five osmotic pressures i.e. 255, 265, 275, 285 and 295mOsm/kg) were used to extend the semen. After freezing, semen characteristics like sperm motility rate, viability rate, acrosomal integrity rate, plasma membrane integrity (PMI) rate, MTT reduction rate, sperm DNA integrity rate and lipid peroxidation were noted. Post thaw sperm motility rate in (%) CEYE was significant (P<0.05) at 295mOsm/kg compared to 255, 265 and 275mOsm/kg. However, sperm motility rate of different osmotic pressures of TEYE and LEYE was non-significant (P>0.05). Sperm viability rate (%) was non-significant (P>0.05) in all three extenders. Sperm acrosomal integrity rate was non-significant in CEYE and LEYE. However, it was significant (P<0.05) at 265, 275 and 295mOsm/kg in TEYE. Sperm PMI rate, MTT reduction rate, sperm DNA integrity rate and lipid peroxidation were non-significant (P>0.05) in CEYE, TEYE and LEYE.
On the basis of the individual and overall comparison of different semen characteristics of three extenders and their osmotic pressures, LEYE with 295mOs.kg was considered to be continued in the next experiment to upgrade the extender by adding taurine (TA) at 0.0, 30, 50 and 70 mM and trehalose (TR) at 0.0, 20, 40, 60 mM concentration. Semen collection, processing, freezing etc were done as per experiment-1 and same post thaw tests were carried out. Post thaw sperm motility rate was significantly (P<0.05) higher at TA-0.0 and TA-20mM and all concentration of TR. Sperm viability rate, acrosomal integrity rate, PMI rate, MTT reduction rate and lipid peroxidation at different concentrations of TA and TR were recorded non-significant (P>0.05). However, sperm DNA integrity rate was significant (P<0.05) higher at TA-0.0 and TR-0.0mM.
On the basis of comparison of different semen characteristics under various concentrations of taurine or trehalose, supplemented in semen extenders. Concentration of TR-70mM was considered to be continued in the next experiment to test fertility of the optimized extender.
Semen straws of LEYE supplemented with TR-70mM were used to inseminate the 50 buffaloes in heat (Supplemented group), while, traditionally used tris based buffalo bull semen extender was used (control group) to compared pregnancy rate (PR) of this experiment. Pregnancy rate in control and supplemented group was 38 and 54% respectively, which was statistically non-significant (P>0.05).
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Isolation, Characterization Of Chondroitin Sulphate And Its Efficacy In Osteoarthritis
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Publisher: 2012 Dissertation note: Chondroitin sulphate (CS) and Glucosamine sulphate (GS) are two main components of articular cartilage. It is believed that these molecules slow down wear and tear of cartilage. Moreover, if administered exogenously as drugs, these may initiate synthesizing capacity of cartilage. Among these, GS promotes the formation and repair of cartilage, whereas CS promotes elasticity and prevent cartilage breakdown by inhibiting degradative enzymes. Concurrent use of both structural units of cartilage as drugs in osteoarthritis (OA) may lessen the progression of disease.
The present study was conducted to elucidate the chicken keel cartilage as an alternate and potential source for this endogenous component that may be used exogenously to repair or prevent damage to joints. Chicken keel cartilages were collected from healthy broilers. CS was extracted using MgCl2 solution (3M), dialyzed and digested with papain. The extracted material was purified by ethanol precipitation, centrifugation and then freeze dried. Proximate analysis of semi-purified polysaccharides revealed the presence of carbohydrates (65.49±0.10), crude protein (12.82±0.26), ash (11.12±.56), moisture (9.88±0.32) and fat (0.69±0.14). Fiber contents were found to be nil in the processed samples. Dimethylmethylene blue binding (DMMB) assay was performed for determination of percent contents of CS in extracted semi-purified samples and mean concentration was found to be 70.77±2.35. Semi-purified polysaccharides were further characterized by FTIR (Fourier Transform Infrared Spectrometer) technique and characteristic Peaks of CS molecules were recorded at 854, 854 and 853 cm-1 and then compared with spectrum of standard CS. Protein content being a major impurity in extracted samples was determined by Bradford method quantitatively (4.64±0.29). Two protein impurities having 77.8 and 50.5 kDa molecular weights were revealed by SDS-PAGE.
Efficacy of semi-purified CS from chicken keel cartilage, standard CS from shark source and GS, alone and in combination in experimental OA rat model was evaluated. To develop OA similar to spontaneous OA, 10mg papain/0.5mL (Sigma, Cat # P 3125) in buffered solution of 0.05 M sodium acetate pH 4.5 was injected intra-articularly in each right knee joint of fifty five albino rats (pre-anesthetized with anesthetic ether). Ten rats (n= 10) were injected with 0.5mL of normal saline (0.9%) in right knee joint that served as control group. Then from fifty five papain injected rats, twenty five were divided into five groups (n=5) for development and assessment of OA model (OA groups). Progression of disease was monitored by clinical scores, histopathological scores and concentration of CTX-II as biomarker in sera samples of experimental rats by ELISA using a commercial kit (serum preclinical CartiLaps ® ELISA kit) for control and OA groups (n=5) on day 0 (control group) and days 1st, 7th, 14th, 21st and 28th post papain injection (OA groups). Highest mean clinical score (10.38±1.1) was observed on 1st day and least on 28th day post papain injection i.e. 5.00±.34. Highest mean histopathological score and CTX-II concentration was recorded on 28th day i.e. 12.82±1.64 and 36.82±3.81. Values of clinical scores, histopathological scores and CTX-II concentration reached to maximum on 21st day and then sustained thereon. Second phase of experiment is comprised of evaluating and comparing the efficacy of extracted CS samples (chicken keel cartilages), standard CS (shark source) alone and in combination with GS. For this purpose, remaining five rats out of ten injected with normal saline intra-articularly served as control groups along with treated and non treated groups of experimental rats. Remaining thirty OA induced rats were divided into six groups (five rats /group). Group 1 (n=5) called non treated group received only placebo till 60th day and served as negative control group.
Treated Group 2 received GS alone, Group 3 CS (standard) and Group 4 were given extracted CS. Group 5 was treated with combination of GS plus CS (standard) and Group 6 with GS plus CS (sample). Doses of glycosaminoglycans (GAGs) were administered as 1.2g/kg/day CS and 1.5g/kg/day GS alone and in combinations. Drugs were offered early in the morning in bolus form with feed (10g) after overnight fasting while non-treated group received only placebo (without any drug).
Anti-arthritis activities of CS standard and extracted alone and in combination with GS were assessed clinically, analyzed statistically by using one way ANOVA. Level of significance (P<0.05) was recorded by using Duncan's Multiple Range (DMR) Post hoc Test. Mean scores of clinical, histopathology and CTX-II concentrations observed at 60th day in control rats (without OA) were 0.00, 0.00 and 2.55, respectively. OA induced untreated group showed mean score for clinical signs, histopathological scores and CTX-II concentrations 4.15, 12.24 and 36.70 and GS treated group 3.19, 3.96 and 6.12 at 60th day of treatment, respectively. For CS (standard), mean scores of clinical signs, histopathological lesions and CTX-II concentrations were recorded as 2.64, 2.44 and 4.48 and for CS (extracted) were 2.26, 2.28 and 4.40 in sera correspondingly at 60th day of treatment. The lowest mean values of clinical signs, histopathology and CTX-II concentrations in sera of treated group with standard CS plus GS were found to be 0.94, 0.94 and 2.62 followed by extracted CS plus GS treated groups 01.05, 1.27 and 2.74, respectively. Clinical, histopathological scores and CTX-II concentrations in group of rats treated with combinations were found to reverse the diseased condition after 60th days of treatment as the values were close to that of normal rats and far away from OA rats. It is concluded that extracted CS from poultry has comparable efficacy with CS standard from shark source alone and in combination with GS. Poultry by-product (keel cartilage) is found to be an alternate and cheap source for CS (chondroprotective agent) as compare to expensive, less available and religiously prohibited source for Islamic countries particularly.
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Seroepidemiology, Zoonotic Potential And Chemoprophylaxis Of Leishmaniasis In Dogs & Human In Pakistan
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Publisher: 2010 Dissertation note: Abstract
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Factors Affecting Biomass Producation Of Baby Hamster Kidney Cell Line In Roller Bottle Culture System For The Production of Foot and Mouth Diseas Virus
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Publisher: 2012 Dissertation note: Foot and Mouth Disease (FMD) is endemic in Pakistan and killed virus vaccines against prevalent serotypes are used for the control of disease. Baby Hamster Kidney (BHK-21) cells as monolayer culture are routinely used for the propagation of FMD viruses. Various nutritional factors: amount of growth medium and serum concentration as well as physical conditions: seeding density, rolling speed, growth time, and incubation temperature for the propagation of BHK-21 cells on roller culture bottles were optimized. The roller culture bottles having surface area of 480 cm2 were used for the propagation of cells. Feeding of cells with 100 ml of growth medium per bottle and supplementation of 5% serum supported the growth of the cells in optimum way. Seeding of ten million cells per bottle resulted into the development of complete monolayer with maximum cell density within 48 hours. The cultured cells remained confluent up to 60 hours while a rapid decline in the number of harvested cells was recorded after 72 hours of incubation. Growth rate of the cells was slower at 33° C that increases at 35° C, reaching to its maximum at 37° C while cells could not tolerate the temperature of 39° C. The bottles kept at rolling speed of 3 rpm yielded maximum amount of cells while higher or lower rotation speed negatively affected the cell proliferation.
Antibody response of buffalo calves to different levels of FMD virus immunogen in trivalent vaccine was investigated. The vaccine containing 106.2 units of immunogen/TCID50 of each of the three serotypes of FMD virus induced log2(1.3± 0.4) units of anti-FMD "O" Complement Fixing Cumulative Geometric Mean antibody (FMD"O" CFT-CGM) titer, log2(1.4±0.3) units of anti-FMD"A" CFT-CGM titer and log2(2.0±0.7) units of anti-FMD"Asia-1" CFT-CGM titer. The vaccine containing 2x106.2 units of immunogen of each of the three serotypes of FMD virus induced log2(2.2±0.2) units of anti- FMD"O" CFT-CGM titer, log2(2.1±0.25) units of anti- FMD"A" CFT-CGM titer and log2(3.4±0.8) units of anti-FMD"Asia-1" CFT-CGM titer. The vaccine containing 3x106.2 units of TCID50 of each of the three serotypes of FMD virus induced log2 (5.3 ± 2.0) units of anti-FMD"O" CFT-CGM titer, log2 (4.6±1.9) units of anti-FMD"A" CFT-CGM titer and log2 (5.0±2.2) units of anti- FMD"Asia-1" CFT-CGM titer. The vaccine containing 4 x 106.2 units of TCID50 of each of the three serotypes of FMD virus induced log2(5.5±1.5) units of anti-FMD"O" CFT-CGM titer, log2(4.4±1.9) units of anti-FMD"A" CFT-CGM titer and log2(5.2±1.9) units of anti-FMD"Asia-1" CFT-CGM titer. Moreover, buffalo calves (n=3) which were primed and boosted with 60 days interval using vaccine containing 2x106.2units of immunogen of each serotype of FMD virus, showed log25.0 and log26.3 units of anti FMD"O"-CFT-GMT antibody titer, log24.6 and log2 6.0 units of anti FMD"A"-CFT GMT antibody titer, log25.6 and log26.0 units of anti FMD"Asia-1"-CFT GMT antibody titer, on 30 and 120 days post boosting.
Each serotype of the virus grew well on BHK-21 cell line. The virus showed poor TCID50 (log 103.2±0.2) in BHK-21 cells having Glasgow Minimal Essential Medium (GMEM) without Fetal Calf Serum (FCS). Addition of FCS in the medium at the rate of one percent increased log 107.1±0.2 units of virus TCID50. Incubation temperature of 350 C and 370 C supported the multiplication and maintenance of BHK-21 cells and yielded log106.6±0.1 and log107.0±0.2units of virus TCID50, respectively. Each serotype of FMD virus showed log106.29±0.07 units of virus TCID50 in the stationary monolayer of BHK-21 cells in Roux flask (75cm2), log107.66± 0.02 units of virus TCID50 in roller bottles (490 cm2) and log108.34 ± 0.07 units of virus TCID50 on 0.2 g of micro-carriers suspending in 200 ml of the growth medium in stirring bottle. The infectivity titer/TCID50 of each of the virus serotypes was significantly higher in roller bottles than that achieved in Roux flasks (p<0.05) and was significantly higher in stirring bottle containing micro-carriers suspending in the growth medium than that of harvested in roller bottle (p<0.05). It is concluded that the infectivity titer of the virus is directly proportional to number of BHK-21 cells in the culture system.
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Characterization Of Mycoplasma Gallisepticum Isolates And Their Use In The Production Of Indigenous
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Publisher: 2012 Dissertation note: Abstract
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Serofpidemiology, Zoonotic Potential And Chemotherapy Of Neosporosis In Dogs And Cattle
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Publisher: 2010 Dissertation note: The aim of current study was to demonstrate the most important features of Neospora caninum infection in Pakistan. In the present study, I examined the prevalence of N. caninum in 7 districts of the country and to accessed the efficacy of various drugs against the parasite in cell culture.
For the achievement of this purpose, the core objectives were,
To have an overview on the overall seroprevalence of neosporosis throughout the country by means of cELISA in aborting, at risk and clinically healthy cows.
To check the correlation of Iscom ELISA and cELISA, and determination of prevalence of N. caninum by means of Iscom ELISA on milk samples.
To identify the transmission of disease towards human.
To determine the efficacy of various drugs against N. caninum.
In phase 1, overall seroprevalence of N. caninum in dairy cattle (detected by means of cELISA, VMRD, Inc., Pullman, WA, USA) was found to be 43.4% with a significant difference (P < 0.05) of seropositivity among all 18 herds (n = 5 aborting herds, n = 13 non-aborting/clinically healthy herds) selected from 7 districts of Pakistan. The seropositivity of cattle to N. caninum antibodies was significantly higher in aborting animals (52.7%) as compared to non-aborting cows ( 41.5%), indicating a significant difference between aborting and non-aborting cattle. In case of pregnant and non-pregnant animals, similar findings were recorded in our study. A significantly higher rate of seroprevalence was observed in pregnant dams (59.8%) than non-pregnant cattle (35.2%). Overall, higher serological prevalence was evaluated during the summer season (61.1%) in all areas followed by autumn (46.9%), spring (34.9%) and least seropositivity was observed in winter season (26.6%).
The difference in seropositivity was significantly different among all age groups, greater in animals older than 2 years of age. Furthermore, the prevalence was statistically significant (P < 0.05) among cattle of different breeds. Seroprevalence in cases of crossbred animals were higher followed by exotic and indigenous breeds.
Phase 2, describes the seroprevalence of N. caninum in clinically healthy dairy cows. A selection of 760 animals from 13 dairy herds located in Punjab and Sindh Province, Pakistan to demonstrate the presence or absence of the Neospora caninum infection in commercial dairy cattle. The serostatus of the cows towards N. caninum was detected by cELISA (VMRD, Pullman, WA). Out of 760 animals, (43.2%) were seropositive to N. caninum. A significant difference of positivity was recorded among all 13 dairy herds. Age wise prevalence though not statistically significant among all age groups, was greater in animals over 2 years of age and least in heifers. Variation was also observed in samples from cattle of different breeds. A significantly higher prevalence was observed in crossbred animals than in purebred and nondescript cattle. Seasonal prevalence was higher during summer season than rest of the seasons. The seroprevalence of N. caninum in pregnant cows was significantly greater than in non-pregnant animals.
Second experiment of this phase describes the seroprevalence of 240 animals from 5 herds with a high rate of abortion, the percentage of seropositivity observed in these herds was 43.8%, slightly higher than the clinically healthy and non-aborting cattle. No significant difference was observed among all sample locations in this experiment. However, significant difference of positivity was recorded among different breeds of cattle. Age wise prevalence, though not statistically significant (P > 0.05), was greater in animals older than 2 years of age.
The assessment of milk samples from lactating cows were also determined for Neospora caninum antibodies by means of Iscom ELISA (SANOVIR® Sanova Biotech AB, Uppasala, Sweden) and showed a good level of agreement (r² = 0.9959) between the two tests (cELISA and Iscom ELISA). Although, the cELISA (VMRD, Inc., Pullman, USA) expressed a higher seropositivity and sensitivity than Iscom ELISA (Sanova Biotech AB, Uppasala, Sweden). Therefore, both of the ELISA tests (cELISA and Iscom ELISA) for the detection of N. caninum antibodies in dairy cattle can perform better in lactating animals. The Iscom ELISA has some advantages over cELISA as it's easy to collect milk samples than serum samples, moreover Iscom ELISA is cheaper and easy to use but has low sensitivity than cELISA and cannot be used in dry animals. The percentage of positivity detected through Iscom ELISA on individual milk samples were 61.4% and 76.6% by cELISA.
In phase 3, an epidemiological study was conducted to determine seroprevalence of N. caninum in dogs of different breeds and age groups. The serum samples of dogs were analyzed by cELISA (VMRD, Pullman, USA) showed a seropositivity of 23.5%. There was no significant difference of seropositivity among various sample locations, highest prevalence was observed in Muzaffar Garh (31.9%), followed by Gujranwala (27.9%), Lahore (25.1%), Hafizabad (20.2%) and least prevalence was recorded in district Okara (14.6%). A significant difference in prevalence of N. caninum antibodies between male (26.1%) and female (18.8%) dogs were recorded. The difference in seroprevalence was not significant among all age groups. The samples with no age record showed a highest prevalence (29.5%) and least seropositivity was observed in adult dogs of 3-6 years of age (18.7%). During Summer season, highest positivity to N. caninum was (31.0%) recorded while the lowest prevalence (16.0%) was observed in Winter season.
Phase 4, describes the seroprevalence and transmission of N. caninum in humans. A selection of 52 serum samples from humans was analyzed for the presence of N. caninum antibodies. The serostatus of the humans towards N. caninum antibodies (IgG) was determined by using commercially available antigen coated IFAT slides (VMRD, Inc., Pullman, Washington USA) and human conjugate. Overall very low prevalence (1.9%) of N. caninum antibodies was reported in this study. Only one case was found to be positive, these findings indicate that no strong evidence of N. caninum infection in humans.
In phase 5, in vitro drug trials was conducted to access the best efficacy of three commercially available drugs. We found that among three anticoccidial drugs i.e Clindamycin, Diclazuril and Sulfadiazine, Diclazuril has best inhibitory effect against N. caninum tachyzoites in cell culture followed by Clindamycin and sulfadiazine.
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Rheological And Microstructural Study Of Commercial Cheddar And Mozzarella Cheeses By Using Farinograph
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Publisher: 2012 Dissertation note: A series of five experiments were conducted using Brabender Farinograph-E to study rheological properties of different brands of cheeses. This is a computerized machine having data recording capacity. It was found that Farinograph was a use full machine for preparing cheese and studying its rheology. The data recorded in the form of Farinogram showed that torque (resistance against flow of farinograph paddles) depended on fat content, temperature employed and time given to cheese formation. Also, the texture of cheese was influenced by these factors.
Sensory tests are not capable of measuring results more accurately as compared to Instrumental tests. To study cheese properties and effects of many manufacturing factors the fundamental methods will help researchers to develop cheeses with required and persistent textural and rheological properties. The instrument most frequently used all over the world for determining water absorption and mixing characteristics of wheat and rye flour in baking industry is Brabender Farinograph®. The present study was conducted by using Farinograph-E as a major tool to measure rheology of cheeses. In this study cheeses of different ages, and kinds e.g., Mozzarella, medium Cheddar, mild Cheddar, old Cheddar, extra old Cheddar, Ricotta and Parmesan were included. The parameters for operating Farinograph-E were developed and initial trials were conducted in various directions to finalize the procedure. Farinograph-E (Brabender GmbH, Duisburg, Germany) was used in this study by using its bowl W-50.
The tests were performed by cutting whole cheese bars into small pieces and shifted into air tight containers. The grated cheese was loaded with the help of spatula into Farinograph bowl. Water bath was adjusted at various temperatures like 20, 30, 40, 50 and 60°C. The temperature was continuously monitored through a temperature probe, inserted into the bowl contained cheese sample. The speed of paddles/ spindles was fixed in Newton meters (Nm) and was kept as constant for all the trials. The lid was closed after filling the bowl and clamped in order to avoid any disturbance. The test was allowed to run for specified time for 35 and 60 minutes. After completion of time durations the test was stopped automatically. The readings were recorded in the form of a graph (torque, time and temperature) of cheese dough resistance over mixing time.
Besides Farinographic studies, the results of Mozzarella and medium Cheddar Farinographic samples of (brand No name) were examined through Cryo-scanning electron microscopy and Fluorescence microscopy to study their microstructure at different stages and their relationship with quality of cheeses. The present study revealed that temperature, time and different fat percentages of different cheese brands shows significant effects on torque values. The results indicated that by increasing fat percentage the torques value decreases.
Cryo-scanning electron microscopy revealed finer details of cheeses. Shape, size and distribution of fat globules were observed through fluorescence microscopy. The changes in globule sizes and their interaction with casein matrix was also observed. Size of globule was estimated using image analysis technique. Aggregation of globules and their rupture was also observed. These changes in fat globules shape and sizes affected flowability, meltability and viscosity of cheeses and thus affected production of torques which were observed in graphs produced by Farinographs.
By studying microstructure it was obvious from micrographs that Stage 1 showed smaller fat globules in large numbers. In Stage 2 the globules became larger in size and lesser in number and like bubbles in shape, as shown in plates. At stage 3, there was no particular change from Stage 2 texture, except slight change in colour. The same changes are depicted in the shape of curve, that moved up and downwards and then upwards.
Full fats at stage 1, showed smaller fat globules those enlarged at stage 2. In stage 3, only enlarged globules were observed, and the resistance increased against paddles of farinograph and sharp increase was seen in the slope of graph. Globules retained their features at next stage and slope in graph became horizontal to x-axis after reaching maximum value.
These results suggest that size distribution of fat globules tended to impose influence on Farinographic results. Overall it is indicated that Farinograph is a suitable instrument for measuring rheology of cheeses.
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Biomass Production Of Pasteurella Multocida By Using Biofermentor For Preparation Of Montanoid Based Vaccine
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Publisher: 2012 Dissertation note: Hemorrhagic septicemia is a contagious bacterial disease of large ruminants principally in cattle and buffalo with high morbidity and mortality. The disease is endemic in nature and outbreaks are common during hot, humid and wet season. The acute and fatal nature and brief duration of the disease limit the antimicrobial therapy. In Pakistan, the disease causes heavy economic losses to dairy industry. Vaccination therefore, is an option for controlling the disease. For a quality vaccine, biomass production of P. multocida along with well developed capsule (immunogen) is necessary. The problem associated with the production of a quality vaccine is poor biomass production of P. multocida when grown in ordinary or routine media.
Present study was designed to isolate P. multocida from sick animals and its molecular characterization in the laboratory and study factors (temperature, media composition, pH incubation time and agitation or shaking) affecting its immunogen production and "in process quality control" factors (biological titer, dry mass, adjuvant and storage time) that affect antibody response. Finally, biomass production of the organism using biofermentor and monitoring of the antibody response of buffaloes to inactivated Montanide ISA-70 based P. multocida vaccine.
Each of the field isolates showed grey, viscous, mucoid, translucent and non hemolytic colonies on blood agar. There was no growth on MacConkey's agar. It was Gram negative coccobacilli or thin rods and bipolar when stained with Leishman's stain. The isolates were positive for Catalase, Oxidase, Hydrogen sulphide and Indole production along with nitrate reduction while it was negative for urease production, citrate utilization and gelatin liquefaction. The bacteria fermented glucose, sucrose, mannitol, mannose, but failed to ferment arabinose, maltose, salicin, lactose, dulcito and inositol.
Polymerase chain reaction (PCR) was performed on isolated colonies by using P. multocida specific and HS causing serotype B specific primers. P. multocida specific PCR gave product of 465 bp while HS causing serotype B specific primers amplified a product of approximately 590 bp.
Growth of the bacteria in casein yeast sucrose broth was optimized under different conditions. CSY broth showed dense growth of P. multocida during incubation for 18 hours. A temperature in between 35°C and 40°C showed its optimum growth. Poor growth was observed below 30°C and no growth was detected at 50°C and above. No growth occurred at pH 0.5 and 10.0 but best growth was obtained at pH 7.0 and 8.0. There was positive correlation between shaking in terms of rpm and growth. There was optimum growth at 500 rpm for 24 hours.
Inactivated HS Vaccine was prepared from dense growth in biofermentor on the basis of dry mass and bacterial count. The effect of biomass, adjuvant, storage of the vaccine, priming alone or with boosting on its potency was also studied along with boosting effect of montanoid ISA 70 oil based vaccine. Dry mass 1.7 mg/dose produced protective antibody titer while bacterial count 10-14/ml was sufficient to produce the protective antibody titer. Montanoid ISA 70 based vaccine provided immunity to buffalo calves better than aluminium hydroxide gel and bacterins. Boosting with oil based vaccine can help to keep the animal immunized for whole year. For better results of vaccine, it can be stored at 4oC for six months.
It is concluded that the proposed study improved quality of the vaccine and reduced volume of the vaccine dose, cost of its production and frequency of vaccination.
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Epidemiology, Zoonotic Potential, Serodiagnosis And Chemotherapy Of Sheep Fasciolosis In Different Ecological zones of balochistan
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; Nature of contents: ; Literary form: Publisher: 2012 Dissertation note: Various epidemiological aspects of human and sheep fasciolosis were investigated in four districts of Balochistan (Pakistan) having different ecology i.e. district Bolan from (Plain zone), Lasbela (Coastal zone), Qilla Saifullah (sub humid and semi arid sub zone of Upland zone) and district Pishin from (Arid sub zone of Upland zone). Sheep samples were examined through Coprological examination showed overall prevalence of 10.26% in one year study period from June 2010 t0 May 2011. The uppermost prevalence was recorded in district Bolan (14.79%) followed by Lasbela (10.63%), Qilla Saifullah (8.75%), and the lowest in district Pishin (6.88%). Overall the highest prevalence by season was recorded in autumn (25.31%) followed by winter (9.22%), summer (6.41%) and lowest in spring (5%). Amongst the month the overall highest prevalence was recorded in the month of September (30.63%) and lowest in the month of May (1.88%). Sex wise prevalence was found highest in female more susceptible to infection (11.22%) than male (8.48), but sex wise difference was non-significant statistically. Amongst the age group significantly higher prevalence was recorded in adults young than adult of age group (5.91%).
During one year study period prevalence (%) of human fasciolosis in some districts of Balochistan was recorded (0.42%), with overall district wise prevalence in Qilla Saifullah and Bolan (0.83%) and (0%) in Lasbela and Pishin. Overall season wise prevalence was noted the highest in autumn (1.25%) followed by summer (0.63%) and 0% prevalence in winter and spring. Month wise results showed 2.5% prevalence only in the month of August and October while 0% in the other months. Gender wise prevalence 0.42% was
found only in male, no female samples were collected due to some religious, traditional and community problems. Prevalence by age was recorded the highest in above 20 years of age group (0.74%) while this value decreased to zero in below 20 years of age group.
Antibodies against fasciolosis in serum samples through indirect (ELISA) were recorded 13.13% (63/480) in sheep and 0.42% (2/480) in human indicates the higher prevalence (%) as compared to fecal examination. Likewise district, age and sex wise seroprevalence (%) of fasciolosis was reported higher than coprological examination in case of humans as well as in sheep.
In sheep positive correlation was noted between fasciolosis and relative humidity while negative correlation with temperature (ºC) and rainfall (mm). While in humans prevalence positive correlation was observed with temperature (ºC), relative humidity (%) and rainfall (mm).
Overall 1123 snails belonging to different 5 genera were collected from different district from different agr-ecological zones of Balochistan from June 2010 to May 2011. Amongst the snails the highest prevalence (37.04%) was found for Indoplanorbis, followed by Bulinus (32.15%), then Lymnea (20.66%), Melanoides (5.52%) and the lowest Physa (4.63%).
Comparative study for coprological and serological tests (ELISA) was conducted for four districts from different agro-ecological zones of Balochistan i.e. District Bolan from (Plain zone), Lasbela (Coastal zone), Qilla Saifullah (sub humid and semi arid sub zone of Upland zone) and district Pishin from (Arid sub zone of Upland zone) for one year i.e. from June 2010 t0 May 2011. Overall prevalence of sheep and humans was 0% and 8.13% by coprological examination and 13.13% and 0.42% by indirect ELISA tests. Prevalence by ELISA was found higher than fecal examination when analyzed statistically. Similar seroprevalence for month, districts, age and sex was noted higher than coprological examination for sheep and humans. ELISA Sensitivity (%) and specificity (%) was recorded >97.0% and 95% and 100%, 100%, respectively for sheep and humans.
Indigenous plants i.e., Saussurea lappa (roots), Fumaria parviflora (aerial) and Caesalpinia crista (seeds) were used at dose level of 60, 70 and 80 mg/kg body weight against naturally infected sheep with fasciolosis and their effectiveness was compared with triclabendazole (10mg/kg body weight). Triclabendazole was found 100 % effective after second dose whereas all herbal medicine it reached up to this mark after administration of second dose of 80 mg/kg body weight. From this study we can conclude that these herbal medicines can safely replace the triclabendazole, which is not, only cost effective but have no side effects.
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Epidemiological, Haematological & Serological Studies Of Leptospirosis In Dogs And Human At High Risk In And Around Lahore City
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Publisher: 2013 Dissertation note: Leptospirosis is an important zoonosis of global importance capable of causing significant subclinical and clinical syndromes both in humans and animals. The disease is characterized by fever, vomiting, diarrhea, myalgia and other signs consistent with renal and hepatic disease. Considering the significance and the substantial losses rendered by Leptospirosis, the present project was designed to study epidemiology and haematology in dogs and humans at high risk in Lahore district and its peri-urban areas.
The study was accomplished in 4 phases. In phase-I, sero-prevalence both in dogs and human was studied including case fatality rate and associated risk factors through cross a sectional study. For this purpose, blood samples were collected from dogs attended at the Pet Centre of University of Veterinary and Animal Sciences, Lahore and other private clinics situated in and around Lahore area through systematic random sampling technique over a period of one year (1st Dec. 2010 to 30th Nov.2011). Blood samples from every fifth un vaccinated dog were collected, but if the dog was vaccinated then the sample was collected from the next unvaccinated one. In this phase 100 sera samples from human volunteers which were at maximum risk (veterinarian, pet and livestock owners, para-vet staff) were also collected. All samples were screened out by using ELISA kits like Canine Leptospira IgG ELISA Kit Catalog no. BG-CAN11485, NovaTein Biosciences, Woburn, MA, USA and Serion Elisa Plate, Virion/Serion GmbH, Wurzburg, Germany for dogs and humans respectively at the Medicine Laboratory and University Diagnostic Laboratory, University of Veterinary and Animal Sciences, Lahore.
To study overall prevalence of Leptospirosis in dogs a total of 429 dogs were examined and it was found that out of 429 blood samples 155 were found positive for Leptospira antibodies. Thus an overall prevalence of Leptospira was recorded as 36.13%. Prevalence of Leptospirosis in dogs during different months of the year was also recorded. The months of September, October and June showed ere the highest prevalence and recorded as 50%, 48.57% and 45% respectively. Although, a few cases were seen during the months of December, January and February while moderate number of cases was recorded during the rest months of the year. There was a significant difference (p<0.05) in the prevalence of Leptospirosis during the different months of the year. Out of these 429, 93 pups and 336 adults were examined for Leptospirosis and found that 26 pups and 129 adults were positive i.e. a prevalence rate of 27.95% (26/93) and 38.39% (129/336) for Leptospirosis was recorded in pups and adult dogs respectively and this difference was non-significant (p>0.05). In this study a prevalence rate of 38.49% (102/265) and 32.31% (53/164) for Leptospirosis was recorded in male and female dogs respectively and this difference between the sexes was also non-significant(p>0.05). According to the results of this study a sero-prevalence of 21.24% (24/113) in winter, 35.82% (24/63) in spring, 40.34% (71/176) in summer and 49.32% (36/73) were recorded in fall season and this difference was significant (p<0.05). The highest prevalence rate was observed in fall and summer seasons of the year during higher rain fall seasons of the year.
To study overall prevalence of Leptospirosis in humans, a total of 100 blood samples were examined through random sampling technique during the whole study period and overall prevalence rate of 44.00% was observed in human population. Different risk factors like different months of the year, age, sex and season were also studied and that the highest prevalence of Leptospira in humans was observed in the months of March, April and August i.e. 66.66%, 66.66% and 60.0% respectively. No significant difference (p>0.05) in the sero-prevalence of Leptospirosis in human during the different months of the year was observed. Sex-wise prevalence rate of 48.71% (38/78) and 27.27% (06/22) for Leptospirosis was recorded in male and female respectively and this difference was significant (p<0.05). The results of this research project revealed a prevalence rate of 47.29% (35/74) and 34.61% (09/26) for Leptospirosis in adults and young ones respectively and this difference was again non-significant (p>0.05). According to the results of this study a sero-prevalence of 41.93% (13/31) in summer, 40.00% (06/15) in fall and 25.92% (07/27) in winter, while 66.66% (18/27) was recorded in spring season of the year and this difference was significant (p<0.05) and the highest prevalence rate was observed in spring.
In phase-II, the effect of Leptospirosis on various blood parameters were determined in both dogs and human. The results of present study revealed a significant difference (P<0.05) between the Hemoglobin (Hb), Erythrocytic sedimentation rate (ESR), Packed cell volume (PCV), Total Leukocytic count (TLC), Neutrophils, Eosinophils and Monocytes of healthy and Leptospira affected dogs, while a non-significant difference was observed (P >0.05)among values of lymphocytes. It showed that values of Hb forthe diseased dogs were lower than healthy ones while ESR, PCV, TLC, Neutrophils, Eosinophils and Monocytes, were more than in normal dogs. Likewise, in humans all the studied parameters were significantly (P <0.05) different between infected and healthy ones. The values of Hb concentration in diseased humans were lower than the healthy ones while ESR, PCV, TLC, Neutrophils, Eosinophils, Basophils, and Monocytes were higher than in healthy people. A negligible change was observed in the percentage count of lymphocytes.
In phase-III, the comparative efficacy of commercially available vaccines against Leptospira was studied. Two commercially available vaccines, Vaccine #1 with protection against two serotypes of Leptospira (Canicola, Icterohaemorragiae) and vaccine #2with protection against four serovars of Leptospira i.e Canicola, Icterohaemorrhagiae, Grippotyphosa and Pomona were compared After six months it was observed through ELISA screening that the vaccine #2 provided better overall protection compared to the vaccine #1 to the pups as well as adult dogs against the Leptospirosis but this difference was non-significant (p>0.05).
In the last phase of this study the chemotherapy trial was conducted. Results found that the efficacy of Penicillin G was 70%, while in group B Amoxicillin produced 60% results and in group C Sarsaparilla proved to be 40% effective against this infection although this difference was non-significant (p>0.05). It is concluded that among therapeutic agents used to treat Leptospirosis in dogs, Penicillin G , Amoxicillin and Sarsaparilla are ranked in respective order of efficacies.
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Epidemological, Serological, Heamatological And Therapeutic Studies On Ovine Nematodiasis In Three Ecological Zones of Balochistan
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Publisher: 2012 Dissertation note: The main area of research in this study was to assess the prevalence, hematological and serological aspects of ovine nematodiasis. Four main experiments were conducted to highlight the objectives of the present research study.
First experiment was conducted to find out the prevalence of sheep major nematodes for one year (January-December 2011). For this purpose three sheep breeds i.e., Balochi, Babrik and Harnai (either sex and between 1-5 years age groups)were selected randomly from three sites i.e., Quetta, Ziarat and Loralai. Faecal analyses of these sheep showed overall higher (40.25%) nematodes prevalence at Loralai followed by Ziarat (29%) and Quetta (23.92%). Five nematodes infection were recorded at three experimental sites. Among these, H. contortus (5.58 to 10.42%)and was the higher prevalent followed by N. battus (6.92 to 9.33%), S. papillosus (4.42 to 9%), T. colubriformis (2.33 to 7.33%) and T. ovis (1.83 to 6.83%).The nematodes prevalence was higher in one and five years old sheep. The female-sheep were infected with higher nematode prevalence higher the than male once and sometimes non-significant difference. These five nematodes were prevalent almost throughout the year; however, a peak infection was recorded during August/September. The high temperature, rainfall and humidity during these months may be predisposing factor of higher prevalence.
Second experiment was on diagnosis of sheep nematodiasis through Enzyme-linked immunosorbent assay (ELISA). For this purpose H. contortus and T. ovis positive samples (200) based on coprological examination were also indicated 100% positive sensitivity by the ELISA based on crude somatic antigen, while on excretory antigen based showed lower (92%) sensitivity. The sera (n=200) of non-infected sheep (based on coprological examination) showed marked difference results. Such as 168 (84%) and 166 (83%) samples were found positive with H. contortus and T. ovis, respectively. While, based on crude somatic antigen 158 (79%) and 144 (72%) samples were found positive with H. contortus and T. ovis, respectively.
Third experiment was conducted to determine the hematological values and total serum protein indices in healthy and nematodes infected sheep. The statistically significant (P<0.05) difference in PCV, Hb, RBC, WBC, Eosinophil, ESR and Total serum protein values was observed among healthy and nematode infected sheep groups. While, there was no statistically significant (P<0.05) difference in TLC, Lymphocytes, Neutrophil, Monocytes and Basophils counts in healthy and nematodes infected sheep groups.
Fourth experiment was conducted on assessing the comparative efficacy of synthetic (Oxfendazole and Ivermectin) and locally manufactured herbal medicine (Deedani, Kirmar and Atreefal Deedan) anthelmintics against sheep nematodes at AZRC/PARC Range-livestock Research Station Sanjavi district Ziarat. The present study results regarding the comparative efficacy showed that, Atreefal deedan among herbal products (Deedani and Kirmar) and Ivermectin than Oxfendazole was found effective against sheep nematodes. The sheep treated with Ivermectin showed highest (96%) FEC reduction, followed by Oxfendazole/Atreefal deedan (86%), Kirmar (60%) and Deedani (32%).
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Prevalence, Identification And Pathogenesis Of Clostridium Chauvoei In Cattle And Buffaloes In Punjab
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Publisher: 2012 Dissertation note: In the first phase of the project, the sampling of diseased animals presumably affected by Black quarter was carried out from six districts of Punjab belonging to three different zones. Around two hundred and fifty samples from each zone were collected and were subjected to bacterial culturing and isolation procedures followed by biochemical identification mechanism. The prevalence of Black quarter in Cattle and buffaloes were thus calculated for each district and zone. Highest prevalence of BQ in Zone II was observed (27.2%) for cattle while in case of Buffaloes highest prevalence (3.2%) was noted in Zone I. similarly higher Prevalence of BQ was noted in 1st quarter of year for Zone I followed by zone II and III while 2nd quarter of season was showing higher prevalence of BQ in zone II and III.
During 2nd phase of experiment tissue samples were inoculated in RCM and blood agar for the re-isolation of C. chauvoei, identified on the basis of colony characteristics and later on subjected to biochemical tests for the confirmation of the isolated organism. Then it was further confirmed through Polymerase chain Reaction for the identification of the causative agent i.e. C. Chauvoei on the basis of 16S rRNA gene sequence. Another set of primers corresponding to alpha toxin gene sequence of C. chauvoeui was also used which strengthened the belief that this strain of C. chauvoei possessed alpha toxin producing ability.
During third phase of project blood samples collected were subjected to hematological estimation for buffaloes and cattle having confirmed as BQ This study revealed significant effect on RBC's count and white blood cells count (P<0.05), while Differential leukocyte count were also showing significant different as compared to Non-infected (P< 0.05). Serum samples were tested for the change in levels of different enzymes. It was found that blood-glucose level and ALT levels were not significantly higher (P>0.05) when compared with control values, Values of AST, CPK and LDH were found significantly higher (P< 0.05) in all infected animals.
Histopathology of affected muscle tissues of both cattle and buffaloes was done to study microscopic changes in the muscle fibers and surrounding tissues. Lesions were somehow disappointing as compared to the magnitude of gross lesions. There were segmental degeneration, Zenker necrosis, discrete edema, occasional neutrophils and emphysema in affected muscle.
Finally, alpha toxin (hemolysin) in culture supernatant of RCM broth was titrated against 2% washed RBC's of cattle, buffalo, sheep, goat, chicken, rabbit and mice to study the hemolytic activity of the toxin. It was found that highest percentage of hemolysis was observed in mice followed by cattle, sheep, buffalo, chicken and rabbit respectively at 25°C. Higher the dilution of toxin, lower the extent of hemolysis. At 37°C variable results were obtained. It showed the biological activity of alpha toxin is also temperature dependant.
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Epidemiology, Zoonotic Potential, Molecular Diagnosis And Chemotherapy Of Cryptosporidiosisin Bovine
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Publisher: 2011 Dissertation note: Cryptosporidiosis is an important parasitic infection of cattle, buffaloes, goats, sheep, horses, cats, human beings and other vertebrates. Prevalence of Cryptosporidiosis in selected animals and human beings carried out on the basis of microscopic examination and polymerase chain reaction (PCR). Percent prevalence of Cryptosporidiosis determined on the basis of conventional identification method was highest in calves (23.1) followed by cattle (10.5) and buffaloes (8.47). Percent prevalence of Cryptosporidiosis in calves, cattle and buffaloes was higher at Government dairy farm (38.33, 20.55 and 16.66) followed by Gawala colonies (26.1, 12.77 and 9.44), Military dairy farm (18.3, 6.11 and 4.44) and then House hold dairies (10, 3.88 and 3.34). Percent prevalence recorded in calves having age less than six months was higher (26.45) than those with 7-12 months of age (16.6). Percent prevalence of Cryptosporidiosis in cattle having age of 2-3 years was higher than those cattle having 3-7 years of age. Similarly, infection rate was higher in buffaloes with 2-3 years age (11.8) than 3-7 years (9.8). Cryptosporidiosis percent prevalence recorded in female calves was higher (24.04) than male calves (18.2). Percent prevalence of Cryptosporidium oocysts observed in feces of male cattle was little higher (11.25) than female cattle (10.4). Cryptosporidiosis percent prevalence recorded in female buffaloes was higher (13.3) than male buffaloes (8.3).
The data was analyzed monthly for the purpose to trace out the specific period of the year having the highest prevalence rate of Cryptosporidium infection. The highest percent prevalence of Cryptosporidiosis recorded in fecal samples of calves was during summer (27.5) followed by autumn (25.8), spring (20.3) and the lowest in winter season (14.5). Overall the highest percent prevalence of Cryptosporidiosis in cattle recorded was during summer (15), followed by spring/autumn (10.88) and the lowest in winter (6.6%). The highest percent prevalence of Cryptosporidiosis recorded in buffaloes was during summer (12) followed by autumn (20), spring (7.5) and the lowest in winter season (4.5). In human beings patients suffering from diarrhea were examined by microscopy and percent prevalence calculated was 40 in present study.
Molecular percent prevalence rate determined was 12.22 in cattle. Percent prevalence recorded using PCR was the highest at Government dairy farm (22.7), followed by Gawala colonies (14.41), Military dairy farm (7.7) and the lowest at House hold dairies (5). The highest season wise percent molecular prevalence was observed during summer (16.6) followed by autumn/spring (13.3), the lowest in winter (7.7). The higher molecular percent prevalence in young cattle (2-3 years) was higher (23.7) than those having age between 3-7 years (10.7). Molecular percent prevalence of Cryptosporidiosis in selected cattle was lower in females (13.6) than males (15).
The efficacy of albendazole observed was 43.05, 58.7 and 64.6 percents on 13th, 20th and 27th day post treatment. The efficacy of albendazole determined on this dose was 34.8, 57.1 and 62.9 percents on days 13, 20 and 27 post therapy. Efficacy of drug calculated on days 13, 20 and 27 was 32.8, 53.3 and 56.6 percent, respectively. Percent efficacy of used drug was 55.04, 68.5 and 79.4 on days 13, 20 and 27 post treatment, respectively. At 50mg/kg body weight dose rate of paromomycin significant decrease in OPG count was recorded from 6th day post treatment and onward (P<0.05). On days 13, 20 and 27 percent efficacy of used drug determined was 48.1, 65 and 69, respectively.
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Characterization Of Linear Type Traits In Nili Rivei Buffaloes Of Pakistan
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Publisher: 2013 Dissertation note: The present study on conformation recording of Nili Ravi buffaloes was planned because there was lack of studies on this aspect of Nili Ravi buffaloes. The main objective of the study was to document and characterize linear type traits in Nili Ravi buffaloes so that the buffaloes with proper body characteristics could be identified for selection and breeding programs. Nili Ravi buffalo herds maintained at Livestock Experiment Station Bhunikey, Pattoki, distt. Kasur, Livestock Experiment Station, Chack Katora distt. Bahawalpur, Livestock Experiment Station Haroonabad distt. Bahawalnagar, Livestock Experiment Station Khushab, distt. Khushab, Livestock Experiment Station Rakh Ghulaman distt. Bhakhar and some private breeders were utilized in this study.
The guidelines for conformational recording of dairy cattle provided by the International Committee for Animal Recording (ICAR) were followed in this study. A total of 437 milking buffaloes were scored for linear type traits on a scale of 1-9. First scoring was performed within 15 to 90 days of calving and then each after about 90 days interval.
Genetic parameters viz. heritabilities, phenotypic and genetic correlations were estimated using Best Linear Unbiased Prediction (BLUP) evaluation techniques. Influencing factors such as age of the buffalo at scoring, stage of lactation, parity, herd and season of scoring were included in the model. Individual Animal Model was fitted under Restricted Maximum Likelihood (REML) Procedure. Data were analysed using the mixed model procedure of the Statistical Analysis Systems. Genetic parameters were estimated fitting an Individual Animal Model using the ASREML set of computer programs.
A total of 1180 records on different linear type traits and body measurements were generated over a scoring period of 2 years. Most of the average values for linear type traits were seen to fall under the intermediate category of 4-6. The means±SD for different linear type traits were found as 5.07±1.35, 5.23±2.35, 5.41±1.45, 5.76±0.98, 6.73±1.53, 4.91±1.85, 4.99±0.88, 4.99±0.90, 5.39±2.13, 4.78±1.1, 5.36±1.56, 4.91±1.84, 5.76±1.67, 3.58±0.88, 5.66±2.24, 6.42±0.88, 4.88±0.69, 4.92±1.08, 4.87±0.84, 5.34±1.79, 4.76±1.78, 5.97±0.94, 5.04±2.488, 5.15±1.65 and 6.44±1.03 for stature, chest width, body depth, angularity, rump angle, rump width, rear legs set, rear legs rear view, foot angle , fore udder attachment, rear udder height, central ligament, udder depth, front teat placement, teat length, rear teat placement, locomotion, body condition score, top line, bone structure, rear udder width, udder balance, teat thickness, thurl width, and temperament, respectively.
A highly significant effect of herd was observed on all of the linear type traits (P< 0.0001). Effect of stage of lactation was found to be highly significant for udder conformation related traits. Parity was observed as a highly significant source of variation for some of the body traits including stature, body depth, body condition score and bone structure. However most of the udder related traits were affected by this factor. A non significant effect of parity was observed on chest width, angularity, rump angle, rump width, central ligament, locomotion, top line, udder balance, thurl width and temperament. A highly significant effect of season of scoring was observed on chest width, angularity, rump angle, rear legs set, rear legs rear view, locomotion and thurl width among body traits. However, stature, body depth, body condition score, top line, bone structure and temperament were not affected by season of scoring. Udder conformation traits including fore udder attachment, rear udder height, central ligament, rear udder width, and udder balance were affected by the season of scoring, however rest of the udder traits including udder depth, front teat placement, teat length, rear teat placement and teat thickness were not significantly different in different seasons.
Significant linear effect of age of the buffalo at scoring was seen on most of the linear type traits. including stature, body depth, rear legs set, rear legs rear view, foot angle, fore udder attachment, rear udder height, central ligament, udder depth, teat length, body condition score, bone structure, rear udder width, teat thickness and thurl width. However, chest width, angularity, rump angle, rump width, front teat placement, rear teat placement, locomotion, top line, udder balance and temperament were not affected by linear effect of age. Quadratic effect of age was found as significant on most of the linear type traits except chest width, angularity, rump width, front teat placement, rear teat placement, locomotion, udder balance and temperament.
Univariate heritability estimates of linear type traits were observed as for stature, 0.36±0.092; chest width, 0.10±0.081; body depth, 0.32±0.081; angularity, 0.06±0.071; rump angle, 0.15±0.071; rump width, 0.38±0.092; rear legs set, 0.02±0.07; rear legs rear view, 0.08±0.07; foot angle, 0.09±0.07; fore udder attachment, 0.21±0.07; rear udder height, 0.09±0.07; central ligament, 0.09±0.09; udder depth, 0.10±0.091; front teat placement, 0.11±0.091; teat length, 0.08±0.091; rear teat placement, 0.11±0.081; locomotion, 0.06±0.06; body condition score, 0.14±0.091; top line, 0.03±0.05; bone structure, 0.09±0.09; rear udder width, 0.15±0.09;
udder balance, 0.16±0.07; teat thickness, 0.22±0.091; thurl width, 0.31±0.09 and temperament, 0.14±0.07, respectively.
Some important positive phenotypic correlations of linear type traits with 305 days milk yield were observed as 0.18±0.04 for body depth, 0.15±0.04 for rump angle, 0.13±0.04 for rump width, 0.30±0.04 for rear udder height, 0.43±0.03 for central ligament, 0.16±0.03 for rear teat placement and 0.19±0.04 for rear udder width. Rest of the phenotypic correlations were very low. Considerable negative phenotypic correlations included -0.16±035 for body condition score, -0.15±0.04 for top line, -0.16±0.03 for front teat placement, -0.14±0.04 for udder depth and -0.26±0.04 for fore udder attachment.
Most of the linear type traits showed positive but low genetic correlation with 305 days milk yield including 0.140±0.0001 with stature, 0.210±0.0001 with body depth, 0.11±0.0001 with rump angle, 0.19±0.0002 with rump width, 0.14±0.0001 with rear udder height, 0.20±0.000001 with central ligament, 0.14±0.0000001 with rear teat placement, 0.13±0.0001 with rear udder width, 0.14±0.0000001 with udder balance, 0.09±0.0001 with thurl width and 0.12±0.0000001 with temperament.
Phenotypic and genetic correlations of most the linear type traits with score day milk yield were generally higher than with 305 days milk yield. Phenotypic correlations with score day milk yield were observed as 0.09±0.03 for stature, -0.21±0.03 for chest width, -0.05±0.04 for body depth, -0.17±0.03 for angularity, -0.12±0.03 for rump angle, -0.16±0.05 for rump width, -0.32±0.03 for rear legs set, -0.16±0.04 for rear legs rear view, -0.22±0.03 for foot angle, -0.34±0.03 for fore udder attachment, -0.16±0.04 for rear udder height, -0.16±0.04 for central ligament, -0.25±0.03 for udder depth, 0.06±0.04 for front teat placement, 0.008±0.03 for teat length, -0.19±0.04 for rear teat placement, -0.15±0.04 for locomotion, -0.22±0.03 for body condition score, -0.35±0.03 for top line, -0.08±0.04 for bone structure, -0.17±0.05 for rear udder width, -0.18±0.04 for udder balance, -0.20±0.03 for teat thickness, -0.11±0.04 for thurl width and -0.11±0.05 for temperament, respectively.
Genetic correlations with score day milk yield were observed as 0.57±0.05 for stature, 0.09±0.02 for chest width, 0.31±0.04 for body depth, 0.06±0.02 for angularity, 0.15±0.03 for rump angle, 0.30±0.05 for rump width, 0.04±0.02 for rear legs set, 0.06±0.01 for rear legs rear view, 0.06±0.02 for foot angle, 0.10±0.02 for fore udder attachment, 0.18±0.03 for rear udder height, 0.12±0.02 for central ligament, 0.18±0.02 for udder depth, 0.60±0.06 for front teat placement, 0.23±0.03 for teat length, 0.07±0.01 for rear teat placement, 0.021±0.02 for locomotion, 0.12±0.02 for body condition score, 0.08±0.02 for top line, 0.08±0.03 for bone structure, 0.19±0.04 for rear udder width, 0.19±0.03 for udder balance, 0.095±0.02 for teat thickness, 0.12±0.02 for thurl width and 0.27±0.05 for temperament, respectively.
Among body measurements, head related measurements included head length, horn diameter at base, length and width of ear and poll width and their average values were found as 54.13±3.48, 18.65±2.06, 29.5±2.12 and 18.66±1.22, and 30.95±2.35 cm, respectively. Average values for neck length and neck circumference were observed as 53.32±4.56 and 95.77±8.58 cm, respectively.
The height and length of body was measured at different body points and average values were found as 139.56±6.29 cm for horizontal body length, 154.01±7.61 cm for diagonal body length, 135.77±4.4 cm for height at sacrum, 132.04±4.57 cm for height at withers, 130.77±4.61 cm for height at 6th rib position, 126.34±4.51 cm for height at last rib position, 128.89±4.83 cm for height at hook bone and 118.81±4.45 cm for height at pin bone.
The average values for heart girth, paunch girth, sprung at 6th rib position and sprung at last rib position were resulted as 194.46±10.31, 238.52±13.96, 45.15±4.48 and 68.72±5.2 cm, respectively. Mean estimates for top wedge area, front wedge area and side wedge area were obtained as 3152.79±309.53, 1030.17±136.34 and 3105.07±345.26 cm2, respectively. The length of tail and its diameter at base was measured and its value averaged 103.51±12.55 and 22.41±2.005 cm, respectively. Average values of skin thickness at neck, ribs, belly and tail region were found as 4.16±1.16, 5.85±1.36, 7.34±1.49 and 1.71±0.55 mm, respectively. Mean values for some other traits included 43.52±2.582 cm for rump length, 3.12±0.56 cm for heel depth and 523.13±81.63 kg for body weight. It was observed that herd was a significant source of variation for all body measurement traits. Age of the buffalo at classification was a significant source of variation for all of the body measurements except horn diameter at base, poll width, tail length, skin thickness at tail and height at hook bone.
Most of the body measurements have been found to be lowly to moderately heritable in the current study. Heritability estimates for various body measurements were observed as 0.16±0.09 for horn diameter at base, 0.38±0.04 for ear length, 0.06±0.09 for ear width, 0.25±0.091 for head length, 0.14±0.09 for poll width, 0.03±0.06 for neck circumference, 0.05±0.07 for neck length, 0.05±0.09 for body length, 0.05±0.09 for diagonal body length, 0.41±0.09 for tail length, 0.28±0.091 for tail diameter at base, 0.04±0.09 for skin thickness at neck, 0.02±0.09 for skin thickness at ribs, 0.10±0.09 for skin thickness at belly, 0.07±0.08 for skin thickness at tail, 0.11±0.09 for height at sacrum, 0.28±0.09 for height at withers, 0.22±0.092 for height at 6th rib position, 0.25±0.092 for height at last rib position, 0.18±0.091 for height at hook bone, 0.07±0.08 for height at pin bone, 0.04±0.06 for sprung at 6th rib position, 0.07±0.06 for sprung at last rib position, 0.13±0.09 for heart girth, 0.05±0.09 for paunch girth, 0.11±0.09 for top wedge area, 0.05±0.06 for front wedge area, 0.16±0.07 for side wedge area, 0.13±0.08 for rump length, 0.02±0.06 for heel depth and 0.33±0.07 for body weight.
Phenotypic correlations of 305 days milk yield with various body measurements were in low range. Positive phenotypic correlations ranged from 0.02±0.04 for sprung at 6th rib position to 0.17±0.05 for ear length. Some of the important body measurements have positive phenotypic correlation with 305 days milk yield as 0.15±0.04 for head length, 0.04±0.04 for diagonal body length, 0.04±0.02 for height at withers, 0.11±0.03 for height at sacrum, 0.11±0.04 for sprung at last rib position, 0.04±0.04 for heart girth, 0.08±0.03 for rump length and 0.07±0.03 for body weight. Negative phenotypic correlations with 305 days milk yield ranged from -0.03±0.03 for side wedge area to -0.25±0.03 for horn diameter at base. Some important negative phenotypic correlations included -0.25±0.03 for horn diameter at base, -0.04±0.04 for neck circumference, -0.12±0.03 for skin thickness at neck and -0.08±0.03 for front wedge area.
Positive phenotypic correlation with score day milk yield included 0.09±0.05 for body weight, 0.07±0.002 for rump length, 0.09±0.003 for sprung at last rib position, 0.09±0.005 for height at hook bone, 0.08±0.02 for height at sacrum. Rest of all the traits were low in correlation with milk yield. Negative phenotypic correlation with score day milk yield included horn diameter at base as -0.15±0.02 and heel depth as -0.13±0.04. Rest of all negative phenotypic correlations were very low.
Positive genetic correlations of 305 days milk yield varied from 0.02±0.002 for ear width to 0.23±0.02 for side wedge area. Some important body measurements have positive genetic correlation values as 0.121±0.000001 for head length, 0.162±0.000001 for diagonal body length, 0.080±0.000001 for height at withers, 0.15±0.000001 for height at sacrum, 0.15±0.000001 for sprung at last rib position, 0.14±0.0005 for heart girth and 0.16±0.007 for body weight. Negative genetic correlation for this trait was observed only for skin thickness at neck region as -0.16±0001.
About 40 traits regarding udder and teat measurements before and after milking were analysed. Average values for udder length, width, height, depth and circumference before milking were found as 52.65±6.87, 53.52±6.19, 54.34±4.99, 18.76±3.87, and 77.05±11.69 cm, respectively while the corresponding values for the same traits after milking were found as 47.08±6.57, 48.15±5.79, 55.39±5.15, 18.11±4.11 and 67.04±8.11 cm, respectively. Teat impression distances between front teats, rear teats, fore and rear teats from right side and fore and rear teats from left side were found as 12.46±3.01, 7.01±1.91, 8.08±1.8 and 7.71±1.75 cm, respectively. Pre stimulation and after milking teat characteristics were found as 12.93±3.12 and 11.71±2.83 cm for distance between front teats; 7.48±1.93 and 6.61±1.58 cm for distance between hind teats; 8.34±1.91 and 7.54±1.60 cm for distance between fore and hind teats of right side; 8.004±1.95 and 7.17±1.60 cm for distance between fore and hind teats of left side; 10.19±2.17 and 9.057±1.50 for diameter of fore right teat; 10.92±2.45 and 9.611±1.66 cm for diameter of rear right teat; 10.33±2.11 and 9.33±1.45 cm for diameter of fore left teat; 11.25±2.54 and 9.937±1.76 cm for diameter of rear left teat; 10.71±2.63 and 11.2±2.39 cm, for teat length of fore right teat; 13.05±3.27 and 13.13±3.03 for teat length of rear right teat; 11.09±2.71 and 11.88±2.61 cm for teat length fore left teat and 13.75±3.04 and 14.47±2.99 for teat length of rear left teat, respectively.
All of the udder conformation traits before and after milking were highly significantly different in different herds (P<0.0001). Stage of lactation was found to be highly significant source of variation (P<0.0001) for before milking udder length, before milking udder height, average before milking udder circumference, after milking udder length, after milking average udder circumference, teat impression distance between fore, between rear and between fore and rear teats on both sides. However, before milking average udder width, before milking udder depth, after milking average udder width, after milking udder height and after milking udder depth were not affected by this factor.
All of the above mentioned traits were significantly affected by parity except after milking udder depth and teat impression distance between fore teats and between rear teats.
Season of scoring significantly affected before milking udder length (P<0.01), before milking average udder width (P<0.05), before milking average udder circumference (P<0.01), after milking average udder width (P<0.01), after milking average udder circumference (P<0.0001), teat impression distance between fore and hind teats of left side (P<0.05). Rest of all the traits were not significantly different in different seasons.
Most of the udder traits were significantly affected by linear and quadratic effect of age of the buffalo at classification. Herd was a significant source of variation for all teat related traits recorded at pre stimulation before milking time. Stage of lactation significantly affected pre stimulation distance between front teats, pre stimulation distance between hind teats, pre stimulation distance between fore and hind teats on right and left side, pre stimulation diameter of fore right teat, pre stimulation teat length of fore right teat, pre stimulation teat length of rear right teat, pre stimulation teat length of fore left and rear left teat. However, pre stimulation diameter of rear right teat, pre stimulation diameter of fore left teat and pre stimulation diameter of rear left teat were not affected by this factor. All of these parameters were affected by parity except pre stimulation distance between hind teats and pre stimulation teat length of fore left teat. Similarly all of these traits were affected by season of scoring except pre stimulation distance between fore, between hind, between right and between left teats.
All of teat characteristics after milking were significantly affected by herd. Stage of lactation significantly affected after milking distance between fore and hind teats of right side (P<0.05), after milking teat length of fore right and rear right teat (P<0.01), after milking teat length of fore left teat (P<0.05) and rear left teat (P<0.0001). Rest of all traits after milking were not affected by stage of lactation. Most of the teat parameters after milking were significantly affected by parity except after milking distance between front and between rear teats, after milking teat length of rear right teat and after milking teat length of fore left teat. Distances among teats after milking and after milking diameter of rear left teat were not significantly affected by season. Rest of all traits were significantly affected by this factor.
Heritability estimates for before milking udder length, average udder width, udder height, udder depth and average udder circumference were found as 0.08±0.07, 0.22±0.08, 0.22±0.09, 0.05±0.06 and 0.21±0.07, respectively. The corresponding values after milking for these traits were observed as 0.14±0.07, 0.20±0.08, 0.09±0.08, 0.02±0.08 and 0.09±0.07, respectively.
Heritability estimates for before milking and after milking teat characteristics were found as 0.11±0.09 and 0.15±0.09 for distance between front teats; 0.03±0.06 and 0.03±0.07 for distance between hind teats; 0.32±0.09 and 0.06±0.07 for distance between fore and hind teats of right side; 0.16±0.08 and 00.09±0.07 for distance between fore and hind teats of left side; 0.21±0.08 and 0.11±0.08 for diameter of fore right teat; 0.05±0.05 and 0.02±0.05 for diameter of rear right teat; 0.19±0.08 and 0.25±0.09 for diameter of fore left teat; 0.07±0.06 and 0.03±0.07 for diameter of rear left teat; 0.12±0.06 and 0.08±0.06 for teat length of fore right teat; 0.02±0.05 and 0.11±0.07 for teat length of rear right teat; 0.29±0.09 and 0.29±0.092 for teat length of fore left teat and 0.14±0.08 and 0.08±0.07 for teat length of rear left teat, respectively.
Phenotypic correlations of before and after milking udder length, average udder width, udder height, udder depth and average udder circumference with 305 days milk yield were found as 0.29±0.04 and 0.18±0.04; 0.30±0.04 and 0.33±0.04; -0.26±0.03 and -0.20±0.03; 0.07±0.04 and 0.06±0.05 and 0.18±0.04 and 0.14±0.04, respectively. Corresponding values in the same order for genetic correlations were observed as 0.17±0.0002 and 0.21±0.0003; 0.33±0.0002 and 0.19±0.0003; -0.29±0003 and -0.34±0003; 0.10±0.0001 and 0.07±0.0001 and 0.28±0.0004 and 0.23±0.0003, respectively.
Phenotypic correlations of before and after milking udder length, average udder width, udder height, udder depth and average udder circumference with score day milk yield were found as 0.29±0.03 and -0.18±0.02; -0.32±0.02 and 0.17±0.01, -0.38±0.001 and -0.20±0.002, 0.28±0.01 and -0.04±0.04 and 0.21±0.04 and -0.15±0.04, respectively.
Phenotypic correlations for pre stimulation and after milking teat characteristics with 305 days milk yield were found as 0.19±0.03 and 0.07±0.03 for distance between front teats; 0.20±0.04 and 0.20±0.04 for distance between hind teats; 0.21±0.03 and 0.21±0.03 for distance between fore and hind teats of right side; 0.18±0.03 and 0.18±0.03 for distance between fore and hind teats of left side; 0.07±0.03 and 0.27±0.04 for diameter of fore right teat; -0.04±0.03 and 0.14±0.04 for diameter of rear right teat; -0.03±0.04 and 0.20±0.04 for diameter of fore left teat; -0.02±0.04 and 0.20±0.03 for diameter of rear left teat; 0.24±0.03 and 0.28±0.03, for teat length of fore right teat; -0.13±0.03 and -0.009±0.04 for teat length of rear right teat; 0.01±0.02 and 0.12±0.03 for teat length fore left teat and 0.06±0.03 and 0.22±0.03 for teat length of rear left teat, respectively.
Genetic correlations for pre stimulation and after milking teat characteristics with 305 days milk yield were found as 0.22±0.0002 and 0.12±0.0003 for distance between front teats; 0.26±0.0001 and 0.13±0.0001 for distance between hind teats; 0.11±0.0001 and 0.09±0.0001 for distance between fore and hind teats of right side; 0.10±0.0001 and 0.07±0.0001 for distance between fore and hind teats of left side; 0.11±0.0001 and 0.11±0.0001 for diameter of fore right teat; 0.09±0.0002 and 0.16±0.0001 for diameter of rear right teat; 0.001±0.000001 and 0.001±0.0001 for diameter of fore left teat; 0.001±0.000001 and 0.001±0.0001 for diameter of rear left teat; 0.080±0.00001 and 0.11±0.0001 for teat length of fore right teat; 0.07±0.000001 and 0.001±0.0002 for teat length of rear right teat; 0.003±0.000001 and 0.003±0.0003 for teat length fore left teat and 0.003±0.000001 and 0.002±0.0002 for teat length of rear left teat, respectively.
Phenotypic correlations for pre stimulation and after milking teat characteristics with score day milk yield were found as -0.37±0.02 and -0.48±0.03 for distance between front teats; 0.04±0.04 and 0.06±0.04 for distance between hind teats; 0.04±0.04 and 0.03±0.04 for distance between fore and hind teats of right side; 0.03±0.039 and 0.08±0.04 for distance between fore and hind teats of left side; -0.33±0.03 and -0.16±0.04 for diameter of fore right teat; -0.46±0.03 and -0.26±0.04 for diameter of rear right teat; -0.41±0.03 and -0.24±0.04 for diameter of fore left teat; -0.30±0.03 and -0.28±0.04 for diameter of rear left teat; -0.43±0.03 and -0.49±0.03 for teat length of fore right teat; -0.36±0.02 and -0.47±0.02 for teat length of rear right teat; -0.41±0.034 and -0.43±0.03 for teat length fore left teat and -0.28±0.021 and -0.53±0.02 for teat length of rear left teat, respectively.
Genetic correlations for before and after milking teat characteristics with score day milk yield were found as 0.13±0.016 and 0.15±0.02 for distance between front teats; 0.30±0.04 and 0.40±0.05 for distance between hind teats; 0.19±0.05 and 0.38±0.05 for distance between fore and hind teats of right side; 0.32±0.06 and 0.44±0.06 for distance between fore and hind teats of left side; 0.22±0.03 and 0.27±0.04 for diameter of fore right teat; 0.16±0.02 and 0.23±0.03 for diameter of rear right teat; 0.15±0.02 and 0.22±0.03 for diameter of fore left teat; 0.11±0.02 and 0.24±0.03 for diameter of rear left teat; 0.19±0.02 and 0.17±0.02 for teat length of fore right teat; 0.075±0.01 and 0.07±0.01 for teat length of rear right teat; 0.27±0.029 and 0.27±0.03 for teat length of fore left teat and 0.10±0.01 and 0.08±0.01 for teat length of rear left teat, respectively.
Least squares means for various performance traits were found as 7.02±2.46 for score day milk yield, 1801.61±624.59 for lactation milk yield, 2074.1±360.85 for 305 days milk yield, 2149.09±680.59 for best milk yield, 272±69 for lactation length, 408.553±203.63 for preceeding dry period, 1762.05±305.97 for age at first calving, 477.68±64.53 for weight at first calving, 110±33 for age at scoring in months, 523.133±81.63 for weight at scoring in Kg.
Most of the phenotypic studies on Nili Ravi breed are limited to recording only few body measurements. In order to explore the physical features of this breed, linear scoring system needs to be adopted which is based on measurement of certain specific parts of body as per international standards according to the ICAR guidelines. However, some of the linear scores developed for dairy cattle breeds do not fit for this breed and harmonization of certain trait definitions is needed even for the linear score system for this breed. The following points are important regarding linear scoring system for Nili Ravi buffaloes:
" In case of rump angle, the score ranging as 1-3 which refers to higher pin bone than hook bone is not present in Nili Ravi buffaloes. The score for central ligament ranging as 1-3 which refers to convex floor of udder has not been observed in this breed. The position of front teat placement as inside of quarter scoring as 7-9 has not been observed in Nili Ravi buffaloes. The position of rear teat placement as outside of quarter scoring as 1-3 has not been observed in Nili Ravi buffaloes. The score for top line ranging as 8-9 which represents a back bent upwards has not been observed in this breed. The score of 1 and 2 which represents a rear udder deeper than the fore udder has also not been observed in the present study. A higher temperament score indicates that buffaloes tend to be excited especially at the time of milking and handling. This behaviour of buffaloes needs to be improved through selection and breeding.
" A highly significant effect of herd was observed on all of the linear type traits. Effect of stage of lactation was found to be highly significant for udder conformation related traits including fore udder attachment, rear udder height, central ligament, udder depth, teat length and rear udder width. Most of the udder related traits were affected by parity such as fore udder attachment, rear udder height, udder depth, teat length, rear udder width and teat thickness. significant effect of parity was observed on chest width, angularity, rump angle, rump width, top line, thurl width, and temperament.
" Initiation of conformation recording in public and private sector and use of selective and planned breeding will be helpful for the improvement in milk yield and to bring uniformity in body features of Nili Ravi buffaloes.
" Scoring in first parity should be adopted as in later parities adjustment for age and parity will be needed.
" Differences among herds for most of the traits suggest that performance can be improved by exploiting genetic potential through selection and breeding. Heritability estimates for most of the linear type traits were found as higher than the reported values available in literature. The reasons might be due to species differences and relatively small data set as well as incomplete pedigree records. Even then the results might be considered for inclusion of some of the linear type traits in selection programs. Keeping in view that this is a preliminary study on genetic aspects of linear type traits in Nili Ravi buffaloes, further studies and research with larger data set is needed to explore linear type traits and to validate the findings of the current study.
" A positive genetic correlation of stature with milk yield suggest that taller and heavier buffaloes produced more milk and selection for taller buffaloes may result in improved milk yield but the efficiency of milk yield must be studied before making indirect selection for milk yield through stature. Negative phenotypic correlation of chest width with score day milk yield suggested that buffaloes with wider chest are relatively less efficient in milk production. Further studies are needed with larger data set to verify the results. A considerable positive genetic correlation between body depth and milk yield suggest that body depth may be considered for indirect selection of higher milk yield in Nili Ravi buffaloes. Considerable genetic correlation with milk yield suggest that rump width is important in this breed of buffaloes and can be used for indirect selection for improved milk yield. A considerable negative phenotypic correlation of fore udder attachment with milk yield is important however negligible genetic correlation suggest that fore udder attachment is independent of milk producing genes and separate selection for each trait should be considered keeping in view heritability of the trait in Nili Ravi buffaloes. A positive genetic correlation of rear udder height with milk yield suggested that selection for this trait might be helpful for improved milk yield in Nili Ravi buffaloes. Genetic correlation of teat length with score day milk yield is considerable in the current study but very low with 305 days milk yield. The findings of current study suggested that rear teat placemen has a considerable genetic correlation with milk yield and can be used for indirect selection for better milk yield. The results of current study are not in agreement with most of the reports in the literature regarding correlation of BCS with milk yield. Further research is needed to verify positive genetic correlation of BCS with milk yield before using BCS as selection criterion for milk yield in Nili Ravi buffaloes. Due to negative phenotypic correlation of body condition score with milk yield, an optimal score of below average ranging from 4 to 5 may be recommended. A positive genetic correlation of rear udder width with milk yield suggested that some of the same genes are controlling milk yield and rear udder width and indirect selection for improved milk yield is possible through selection for rear udder width in Nili Ravi buffaloes. This genetic correlation with milk yield is considerable but further studies are needed before the udder balance could be included for selection program in Nili Ravi buffaloes.
" Current study indicated that teat thickness is not genetically important with negligible correlation with milk yield in Nili Ravi buffaloes but negative phenotypic correlation is considerable and buffaloes with thinner teats are suitable for more milk production. A low but positive genetic correlation of thurl width with milk yield provides a scope for further studies to explore this trait in Nili Ravi buffaloes. Further studies are needed with relatively larger data set to explore temperament and verify its relationship with milk yield in this breed of buffaloes. Generally, the least squares means for most of the body measurements were found in the normal range and were in agreement with most of the reports in literature.
" Comparatively higher body weight was observed than the reports available for Nili Ravi buffaloes. One of the reason for this might be relatively better supply of feed and fodder during the course of study and also the records pertaining to 3rd and latter parities were more in number than the records on younger buffaloes. The top and side wedge area are almost similar with less variation showing that Nili Ravi buffaloes are relatively more wedge shaped.
" Most of the body measurements were affected by the herd and age factors but the effect of parity, stage of lactation and season of scoring was variable for different traits and showed not very clear trend. Body weight was affected by all the factors studied in the current investigation. Most of the body measurements have been found to be moderately to highly heritable in the current study. Overall range of heritability estimates for body measurements was found as 0.08±0.09 to 0.92±0.00.
" Skin thickness has been found under the genetic control and can be improved through selection and breeding keeping in view its importance and demand in the leather industry and also its correlation with milk yield.
" Diagonal body length in the current study has shown a low but positive genetic correlation with milk yield and this trait might be considered in the selection program for Nili Ravi buffaloes. The negative genetic correlation of skin thickness in the neck region with 305 days milk yield is important and advocates the thinking of farmers about the negative correlation of skin thickness with milk yield. Genetic correlation of heart girth with milk yield although not very high but seems to be important and can be considered for indirect selection for milk yield through heart girth measurement. A reasonable genetic correlation of body weight with milk yield suggested that this trait should be considered in the selection program for improved milk yield in Nili Ravi buffaloes.
" Udder colour has not been found important. Buffaloes with pendulous udders have produced more milk. The possible reason for this more milk is that such buffaloes were recorded in latter parities and age of those buffaloes was high and the size of their udder was large. The frequency of buffaloes with such type of udder is only 8%. Buffaloes with such type of pendulous udders are more prone to udder and teat injuries and mastitis and their life time production is less. Thick and lengthy teats have been observed in this breed and the reason might be due to hand milking and direct suckling of cows by the calves.
" Most of the udder traits were significantly affected by herd, parity, stage of lactation and age of the buffaloes at classification. Most of the udder measurements have been found highly heritable and this provides a good scope for improvement of these traits through selection and breeding. A general decrease in the distance between fore, rear and fore and rear teats on both sides was observed after milking. This indicated that the distance measured after milking was a good indicator of actual distance between teats of this breed irrespective of stage of lactation. Udder length, width, udder circumference and height either recorded before milking or after milking have been found genetically correlated with milk yield and they should be considered for selection decisions in Nili Ravi buffaloes. A reasonable positive genetic correlation of distance between fore and between rear teats suggested that this distance is important for milk yield and should be considered for selection in Nili Ravi buffaloes. The results of present study suggest that teat diameter is not genetically much important for milk yield and the reason of thick teats is due to hand milking and direct suckling by the calves.
" Teat distance between front teat, between rear teat, diameter of fore right and rear right teat and teat length of fore right teat have shown low but not negligible genetic correlations with milk yield and should be given some importance in making selection decisions in Nili Ravi buffaloes.
" Brown colour buffaloes have not been observed in this study because such animals at Govt. livestock farms are culled at an early age, however farmers think that such type of buffaloes are better milk yielder and they like and demand such animals, development and conservation of these animals is advocated at experimental level to study their potential.
" Further research is needed to evaluate visual image analysis system as a tool for quick and more accurate conformation recording.
Availability: Items available for loan: UVAS Library [ Call number: 1708,T] (1).
Production And Evaluation Of Peste Des Petits Ruminants Virus Vaccine
Material type: Book ; Format:
; Literary form:
Publisher: 2013 Dissertation note: PPR is an acute highly contagious viral disease of small ruminants which is endemic in Pakistan. Present study was aimed to evaluate the freeze dried PPRV vaccine with variable biological titer to induce protective immune response in beetal goats. The comparative immune response of animals to adjuvant and non-adjuvant vaccine and duration of immunity was also studied. Effect of boosting on the humoral immune response of animals as well as shelf life of various vaccines was also evaluated. Each of the vaccines was inoculated in a group of five animals. Serum samples were collected at specified time intervals and antibody levels were detected through cELISA as PI values and neutralization test as MNA titer.
The virus was propagated on the Vero cells. It was estimated that infecting 2 x 107cells with 104.00 TCID50 virus concentrations added to a T-175 cell culture flask at the time of subculture yielded maximum virus titer in the cell culture harvest following three freeze thaw cycles of the contents. The freeze dried vaccine with a biological titer of 105.00 TCID50 per dose provoked maximum antibody titer followed by the ones with a titer of 104.00 or 103.00TCID50 which provoked nearly equivalent protective immune response while the animals inoculated with a vaccine having 102.00 TCID50virus concentrations developed minimum antibody titer. The oil adjuvant PPRV vaccines elicited significantly higher antibody titer in comparison to gel based vaccines but however minimum antibody titers were detectable in response to freeze dried vaccines. Although protective antibody level (? 10 neutralizing antibody units) was detectable in the animals vaccinated with either oil based, gel based or freeze dried vaccine containing biological titer of 104.00 TCID50 but however the extent and duration of immunity was found to be most superior in response to oil based vaccines. It can be concluded that a single shot of either gel or oil based vaccine can provide protection in the vaccinated animals for a minimum of one year duration. Goats receiving a booster dose of the vaccines had a significantly higher antibody tier in comparison to the ones who received single dose of the vaccines. The freeze dried and wet vaccine kept at 4 °C did not show any significant drop in the biological activity of the virus even after 12 months of storage. Immunogenicity of the both adjuvant and non-adjuvant vaccines, as measured through the immune response in the vaccinated animals, also remained unaffected after 12 months of storage at 4 °C.
Availability: Items available for loan: UVAS Library [ Call number: 1731,T] (1).
Improving Nutritional Value And Acceptability Of Dairy Products With Lower Contents Of Saturated Fatty
Material type: Book ; Format:
Publisher: 2013 Dissertation note: Abstract
Availability: Items available for loan: UVAS Library [ Call number: 1742,T] (1).
Survival Of Probiotics In Yogurt Ice Cream
Material type: Book ; Format:
; Literary form:
Publisher: 2013 Dissertation note: This study was designed to produce the yogurt ice cream containing probiotic
microorganisms with the recommended levels (106-107) of live cells at the time
consumption. The mixture was supplemented before freezing with prebiotics (inulin
and oligofructose) and cryoprotectant (glycerol) to see their (prebiotics and glycerol)
effects on the survival of Streptococcus thermophilus, Lactobacillus bulgaricus,
Lactobacillus acidophilus and Bifidobacterium lactis during freezing process and in
storage period. Along with bacterial population, the effects of prebiotics and glycerol
supplementation on physicochemical properties like air holding capacity, fat
components, protein contents, total solids, hardness, stickiness, melting rate, glass
transition, air cell size and ice crystal size were also investigated. Glass transition
temperature was analyzed in each treatment mixture before freezing with differential
scanning calorimeter. The results from the data obtained at various stages of study
have shown different variation pattern for each property.
The initial count of each bacterium before freezing in all treatments with in
experiment was similar and during the freezing process there was non-significant
change in bacterial population. During the storage period at -20°C in the first three
weeks there was less loss in all the samples (P<0.05). With the passage of time the
death rate is increased in all the samples but this decrease was very less with
supplementation as compared to control samples (P<0.05). In the prebiotic yogurt ice
cream the greatest loss was observed in L. acidophilus (P<0.05), while the S.
thermophilus concentration was the maximum among all the bacteria (P<0.05). At the
end of 12 weeks storage period all the bacteria maintained the minimum required
(106-107 CFU/g) concentration (P<0.05). The addition of prebiotics and glycerol has
increased the total solids of all the samples (P<0.05) that would ultimately led to
more overrun percentage. The supplementation of prebiotics and glycerol have shown
non-significant effect on the fat quantity while decreased the protein concentration
significantly (P<0.05). Fat and protein contents remained unchanged during the
storage period of 90 days (P<0.05). The hardness increased with prebiotics and
decreased with glycerol supplementation, while the stickiness increased with the
increasing prebiotics and glycerol concentration (P<0.05). The melting rate has
shown different behavior although the dry matter contents increased with prebiotics
and glycerol but it did not support the slow melting (P<0.05). Prebiotics show less
effect on glass transition temperature, the increase was very less almost to 1°C.
Glycerol has shown most of the effect and it decreased Tg to near about 10°C in 4%
supplemented samples (P<0.05). The overrun percentage show most of its effect on
probiotics as these bacteria are anaerobic and grow best in absence of oxygen, but the
addition of glycerol minimized its effect on survival rate of the bacteria. The overrun
have shown no effect on total solids, fat and protein level but it decreased the melting
rate at 22 °C. The air act as insulator and prevent the melting of yogurt ice cream
(P<0.05). The hardness and stickiness also decreased with increasing level of overrun
(P<0.05). The prebiotics and glycerol supplementation have shown non- significant
change in air cell size and ice crystal size while overrun percentage has significantly
decreased the air cell and ice crystal sizes (P<0.05).
In conclusion, the addition of prebiotics and glycerol increased the survival
rate by decreasing the freeze damage caused by large ice crystal formation and also
improved the physicochemical properties of yogurt ice cream.
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Immuno Pathological Effects Of Neem (Azadirachta Indica) In Commercial Broiler Chickens
Material type: Book ; Format:
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Publisher: 2012 Dissertation note: These experiments were conducted to study the effects of Azadirachta indica admixed in poultry feed on weight gain performance, haemtological values,immune modulations, and toxic effects in broiler chickens. A total number of 144 commercial broiler 1-day old chicks were reared in the experimental sheds of the Department of Pathology, University of Veterinary & Animal Sciences, Lahore, The birds were fed with balanced commercial feed and water ad libitum. The birds were divided into 3 groups; A, B and C having fourty eight chicks each. Birds of all groups were sub divided into four groups of each i.e. A1, A2, A3 and A4; B1, B2, B3 & B4 and C1, C2, C3 and C4, respectively. Each of the sub groups containd 12 birds. Sub groups A4, B4 and C4 were control group with no medication. The birds of groups A, B and C were fed with poultry feed containing dry powder of neem leaves @ 2 gm, 4 gm and 6gm per kg of feed respectively. The birds of groups A1, B1 and C1 were treated with the herb from day 0 to 42 of their life. The birds of groups A2, B2 and C2 were given the neem from day 14 to 42 of their life, whereas the birds of groups A3, B3 and C3 were treated with the herb from day 28 to 42 of their life. Difference between weekly weight gain in the birds of groups A1, B1 and C1 was non significant (P>0.05) however the difference between weight gain in the treated and control groups was significant (P<0.05). The birds treated with the herb from day 0 of their life showed more weight gain. There was no difference in the haematological indices between all of the treated groups and the control groups. The neem treated birds showed increased antibody titers against ND and IBD viruses as compared to control groups. The values of ALP and ASTshowed decreasing trend when the level of neem leaf meal was increased in the ration. Serum creatinine and serum uric acid values posed a slight declining trend in the neem fed birds. There was a decrease in serum cholesterol level in the neem treated bird groups, the higher the concentreation of the herb, the lower the cholesterol value. The organ body weight indices showed that there was no significant difference in liver, spleen and thymus weights among treated groups as well between treated and control groups. There was absence of prominent gross pathological lesions in liver, spleen, kidneys and thymus, however some treated groups showed mild hypertrophied liver and kidneys as did the organs of the birds in control groups. No histopathological changes except a few mild changes were observed in liver, spleen, kidneys and thymus in the birds of experimental groups.
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