Genotyping Of Echinococcus Granulosus And Its Comparative Prevalance In Camels And Human Beings
Material type: Book ; Format:
Publisher: 2008 Dissertation note: Hydaiidosis is caused by metacestode of the dog worm Echinococcus granulosus. It is a serious problem br both Public health and livestock economy. Echinococcus granaiiosu.s has number of genetically distinct strains which are known to differ morphologically and epidemiologically. Out of 100 camels examined only 25 Samples of hydatid cysts were collected from different organs i.e. livers, kidneys, lungs and hearts from Lahore abattoirs. Fertility and viability of the cysts was observed microscopically. Genotyp ing of Echinococcus granulosus was performed through Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR-RFLP). Seroprevalence of hydatidosis in 25 butchers working in abattoir was also determined by the use of Latex agglutination test (LAT) kit for detection ob hydatidosis.
Considerable information is available about genetic variants of E. granulosus around the world. Ten genotypes of E. granulosus have been described, which exhibit a diversity of morphology, development, and host range, as contrmed by various studies. In the Mediterranean area, the CI or common sheep strain, G2, Camel strain G6, and the equine strain G4 have been found in Spain, Italy, Lebanon, and Syria
To date, molecular studies using mainly DNA sequences have identitied G-6 strain of E. granulosus. This categorization follows very closely the patterns of strain variation emerging from biological and epiderniological traits.
In this study we perform serum analysis of butchers to detect antibodies against Echinococcus so that the prevalence of Echinococcus can be checked; the data available indicated that 14% of butcher's population is infected with Echinococcus. In order to confirm the strain of Echinococcus in camels the PCR-RFLP analysis were performed. The data obtained was analysed and it was concluded that the G6 strain of Echinococciis is prevalent in camels in Pakistan. The results demonstrated that PCRRFLP analysis of samples of patients suspected for Echinococcus is a promising diagnostic method and also confirms the type of Echinococcus prevalent in that area and also enables an early direct detection of parasite DNA. This will help to curtail this drastic malady at an early stage and will help to devise the strategy to minimize the losses due to this disease.
It is hoped that the findings of the present study will be helpful for further planning about the control of the disease and correlating the prevalence in camels and butchers from the zoonotic point of view.
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Molecular Detection And Speciation Of The Canme Piropiasm
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Publisher: 2008 Dissertation note: An epidemiological study of babesiosis in dogs was conducted at Pet center, University of Veterinary and Animal Sciences, Lahore, for one year and information on age, sex and breed was gathered. It was found that from a total number of 6204, dogs up to two years of age were more susceptible than other age groups (2-4, 4-6 and above 6 years).The data regarding genders revealed that males were more prone to the disease than female dogs. As far as the breeds were concerned, crossbred dogs were more susceptible followed by Pointers, Alsatians, German shepherds and Bull terriors.Hot and humid months (June to September) have greater impact on the occurrence of disease. The study regarding identification of ticks revealed that Rhiphicephalus sanguinus is the predominant vector of the disease in Pakistan.
Molecular studies were conducted to characterize and identify the species responsible for canine babesiosis in Pakistan. In this regard, a nested polymerase chain reaction-Restriction fragment length polymorphism (PCR-RFLP) technique was employed on different specimens (Blood, Body tissues and Ticks). For this purpose blood samples were collected from twenty four chronically infected dogs and applied on the Flinders Technologies Associates (FTA) cards for transportation to Australia. Different body tissues (Liver, Spleen, Kidney, Intestine, Bone marrow and Pancreas) were procured after euthanizing the two dogs and DNA was extracted, for further studies. Similarly, the eighty eight ticks were also collected from the infested dogs in the 70% ethanol for transportation to Australia. A nested PCR-RFLP assay was used for the detection and differentiation of Piroplasm species on the basis of the 1 8S ribosomal RNA gene. The assay potentially amplified and identified Babesia gibsoni as the main canine piroplasm. Similar assays on the DNA extracted from body tissues and ticks revealed Babesia gibsoni as the main piroplasm. The PCR was found to have a high detection limit (equivalent to i0 dilution), when using the DNA extracted from blood applied to FTA cards, body tissues and ticks. A new technique was developed for extraction of DNA from FTA cards and tick, in this technique, instead of using the FTA specified punching machine, we used scalpel blades, and so the rest of the chemicals used are'generally and easily available. The same protocol was used for extraction of DNA from ticks, only chemicals used in different quantities with different spinning times. Both of which, resulted in cost reduction, less effort and speedy DNA extraction. The technique reported here has the potential to be standardized for routine DNA extractions from FTA cards and ticks.
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Epidemiology, Molecular Diagnosis And Chemotherapy Of Giardiasis In Bovine
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Publisher: 2009 Dissertation note: Giardia is a protozoan parasite of the small intestine that causes extensive morbidity worldwide. Dairy calves can excrete high numbers of the cysts of Giardia and the disease in cattle is clinically important and can reduce the growth performance of the ruminants. Giardia is the cause of non-viral diarrhoea in humans and is responsible for epidemics in the developed and developing countries. The cyst is the infectious form, is ingested in contaminated water or food or directly from faecal-oral contact. Giardia duodenal is the only species, which is found in both humans and animals including dogs, cats, bovines, pigs, sheep and equine.
The present study was conducted to determine the prevalence in bovines at Military dairy farm, Gawala dairy colonies, the Government dairy farm and Household dairies in Lahore. The effect of season, sex, and age on infection rate and shedding of the cysts were also noted, and association of the Giardia infection with normal and abnormal stools was also studied.
Overall 2160 bovine faecal samples (720 buffaloes, 720 cattle and 720 calves) were examined during the study period from August 2007 to July 2008, amongst calves 362/720 (50.27%) were found to be positive. The highest prevalence was recorded in the Government. Dairy farm (68.33%) followed by Gawala colonies (55%), then the Military dairy farm (44.33%) and the lowest (34.44%) was recorded in Household dairies. Overall, highest (61.6%) seasonal prevalence was recorded during autumn, followed by spring (60.83%), then summer (53.4%) and the lowest
(34.1%) was recorded during winter. The highest (65%) prevalence was reported during August and the lowest (3 0%) during December.
Females were found to be more susceptible (56.74%) than males (35.1%). The prevalence was significantly higher (71.52%) in younger calves than the adults
Overall prevalence in cattle was 28.05%. The highest (41.67%) prevalence was recorded at the Government dairy farm, followed by Gawala colonies (32.72%), then the Military dairy farm (22.72%) and the lowest (15%) was recorded in Household dairies. The highest (35%) prevalence was found during August and the lowest (21%) during January. A significant difference (P<0.05) was noted. Females were found to be more susceptible (29.21%) than males (18.75%). The young calves had significantly higher (3 8.88%) prevalence as compared to the adults (24.44%).
Similarly, the overall prevalence in buffaloes was found to be 20.11% percent. The highest (40.55 %), prevalence was recorded at the Government Dairy Farm, followed by Gawala colonies (30%) then Military Dairy Farm (21.11%) and the lowest prevalence i.e. 12.77% was reported in Household Dairies. A non significant difference was recorded P>0.05). The highest (46.66 %) prevalence was recorded during August, while, the lowest (6.66%) during November and December. Females were found to be more susceptible than males. Where as the prevalence in a younger buffalo was significantly higher as compared to the adults.
Comparison of direct microscopic examination and PCR based methods was made at the Government dairy Farm, Gawala colonies; Military Dairy Farm and
Household Dairies. By direct Microscopic examination prevalence was found to be 28.05% (202/720) in cattle whereas by PCR it was 31.11%. Statistically analysis showed that the prevalence by PCR was significantly (P<0.05) higher than the microscopic examination.
It was observed that the highest prevalence of Giardiasis in bovines (Calves, Cattle and buffalo) was noted during August when the average temperature was 31.48°C. However the maximum and minimum temperatures were 35.37°C and
27.6°C, relative humidity 7 1.28% and rainfall was 3.2mm. The results of therapeutic trials by using albendazole, metronidazole, and mebendazole in cattle were calculated on the basis of reduction in the cysts count in the faeces after treatment.
Efficacy of albendazole at three dose levels i.e. 1 Omg/kg.b.wt, 1 5mg/kg.b.wt, 2Omg/kg.b.wt was 86.33%, 98.5% and 100% respectively, on day 27 after treatment. Efficacy of the metronidazole at 5Omg/kg.b.wt, 1 OOmg/kg.b.wt, and 1 5Omg/kg.b.wt. Was 85.42%, 87.8% and 94.02% respectively on day 27. Efficacy of mebendazole at three dosage level i.e. 7.5rng/kg.b.wt, lOmg/kg.b.wt and 2Omg/kg.b.wt was 81.15 %, 87.32%, and 90.4% on day 27 after treatment. Among these drugs, albendazole at
1 5mg/kg.body.weight was found to be most effective drug in the elimination Giardia infection. The significant (P<0.05) decrease in the CPG count after treatment in all the three groups and dose levels was noted. A significant difference (P<0.05) was observed in the level of leukocytes and of eosinophisl of infected cattle at day 06 and day 13 post inoculation. The leukocytes/lymphocytes count of Giardia infected cattle was 58.09%. Whereas, eosinophils constituted of leukocytes 9.69%. The total proteins of the sample were studied by sodium doedocyl sulphate polyacrylamide gel ELECTROPHORESIS (SDS PAGE). The result indicated that 8 diffeent molecular weight peptide badns were identified with size ranges from 20 to 70 KDa and common bands reported at 20, 24 and 35 K Da
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Studies On Rats And Mice As A Reservoir Of Zoonotic Parasites
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Publisher: 2009 Dissertation note: A total of 3600 rats and mice were examined for the prevalence of various zoonotic parasitic infections. Species of various parasites were identified. The order of ifection rate was as Hymenolepis nana (59.9%), Toxoplasma gondii in rats and mice 52.0%), Hymenolepis diminula (35.4%), Cryptosporidium parvum (16.5%), Taenia taenieformis (11.7%), Toxoplasma gondii in humans (11.3%), Capillaria hepatica 7.0%), Babesia spp. (2.3%), Trypanosomu spp. (2.3%) and Leishmania spp. (1.2%).
The highest (78.0%) month-wise prevalence of FL nana was found during August whereas the lowest (43.0%) during February. The highest (70.8%) season-wise prevalence was noted during summer followed by autumn (62.9%) then spring whiles the lowest (46.8%) during winter. Infection was higher in males (78.0%) than females 45.9%). Adult rats and mice were more commonly affected than younger. Infection was higher in rats (65.1%) than mice (16.3%).
The highest (48.0%) month-wise prevalence of H diminuta was found during August whereas the lowest (28.0%) during January. The highest (45.4%) season-wise prevalence was noted during summer followed by spring (35.1%) while the lowest (29.3%) during winter. Infection was higher in males (43.8%) than females (29.3%). Adult rats and mice were more commonly affected than younger. Infection was higher in rats (37.3%) than mice (20.2%).
The highest (23.0%) month-wise prevalence of C. parvum was found during July whereas the lowest (10.0%) during December. The highest (22.4%) season-wise valence was noted during summer followed by spring (16.9%) then autumn (15.3%) le the lowest (11.3%) during winter. Infection was higher in females (19.9%) than es (12.0%). Adult rats and mice were more commonly affected than younger, infection was higher in rats (17.7%) than mice (7.6%).
The highest (20.0%) month-wise prevalence of T raenieformis was found during y and August whereas the lowest (6.0%) during December. The highest (18.4%) son-wise prevalence was noted during summer followed by autumn (10.8%) then ring (10.6%) whereas lowest (7.0%) during winter. Infection was higher in females 15.6%) than males (6.5%). Adult rats and mice were more commonly infected than younger. Infection was higher in rats (12.4%) than mice (6.0%).
The highest (12.7%) month-wise prevalence of C. hepatica was found during August whereas the lowest (4.0%) during November. The highest (10.8%) season-wise prevalence was noted during summer followed by autumn (6.0%) then spring (6.2%) hues the lowest (4.9%) during winter. Infection was higher in females (8.4%) than males (5.0%). Adult rats and mice were more commonly affected than younger. Infection higher in rats (7.3%) than mice (4.6%).
The highest (9.0%) month-wise prevalence of Babesia spp. was found during August whereas infection was absent during January, February, March, April, November and December. The highest (5.0%) season-wise prevalence was noted during summer followed by autumn (4.2%) then spring (0.1%) and was absent during winter. The infection was higher in males (2.7%) than females (2.0%).The infection was present only in adult rats and mice and no case was recorded in younger. Higher infection was noted -n (2.3%) and than mice (2.4%).
The highest (7.3%) month-wise prevalence of Trypanosoma spp. was found August whereas it was absent during January, February, November and December. The highest (5.7%) season-wise prevalence was noted during summer by spring (2.2%) then autumn (1.2%) and it was absent during winter. The infection was higher in females (2.5%) than males (2.0%). Adult rats and mice were more commonly affected than younger. Higher infection was noted in rats (2.5%) than mice
The highest (4.7%) month-wise prevalence of Leishmania spp was found during y and August whereas infection was absent during January, February, March, April, October, November and December. The highest (3.4%) season-wise prevalence was during Summer followed by autumn (1.1%) then spring (0.1%) and infection was during winter. The infection was higher in females (1 .4%) than males (0.9%). The infection was found only in adult rats and mice and no positive case was recorded in 1younger. The infection was (1.2%) in rats and mice.
The role of meteorological data i.e temperature, humidity and rainfall was also studied. It was noted that there was a positive co-relation of high temperature and humidity with the prevalence of various parasitic infections. As the infection were maximum during summer when the temperature and humidity was favorable for high prevalence of parasitic burden.
This study will he helpful to disseminate information regarding zoonotic potential parasitic infections via media i.e. newspapers. magazine, electronic media (T.V, I.T, Radio) through seminars, meetings and conferences with professionals doctors, paramedical staff and livestock farmers.
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Epidemiology, Serodiagnosis, Therapy And Control Of Schistosomiasis In Buffloes
Material type: Book ; Format:
Publisher: 2008 Dissertation note: Schistosomiasis is one of the major diseases of public health and socio-economic importance in the developing countries of the world. Among human parasitic diseases, Schistosorniasis ranks second to malaria in tern-is of world wide public health risk. Keeping in view the importance of disease, the study was conducted to record the month wise and season wise prevalence of Schistosorniasis in buffaloes in four districts of the Punjab, province ie., Lahore, Sargodha, Kasur and Sheikhupura. The present study comprises of four parts.
Part I: deals with epidemiology of Schistosomiasis in buffaloes. Overall prevalence of Schistosomiasis in buffaloes, different farms of Punjab province indicated that infection was the highest (17%) at Kasur district followed by Sheikhupura (14.83), then Lahore (14.6%) and the lowest (13.66%) at Sargodha. The highest month wise prevalence was recorded during August (25.5%) followed by July where as the lowest during December and January.
Infection in buffaloes was higher in animals over two years of age (1 9%) than animals below two years of age (5%) in all the four districts of Punjab. The prevalence was higher in females (15.98%) than male (9.48%). There is variation in the prevalence as there is difference in the environmental and managemental condition of the area. For the serodiagnosis i.e. ELISA was used, the results indicated that the prevalence was lesser than the faecal examination because this was more specific and sensitive than the faecal examination.
Part 2: deals with the prevalence and ecology of snails. Various species of snails which act as the intermediate host of the Schistosomes were collected from the study area. The e of infection in the snails and role of cercariae in transmission of the disease was studied. A total of 10418 snails were collected of these 13.51 per cent were found to be infected. Among these 2350 were collected from Kasur district with infection rate of 14.51 percent followed, by Sheikhupura 2882 (13.6%) then Sargodha 2709 (13.40%) and the lowest at Lahore 2477 (12.51%). At Kasur district, genus wise prevalence of snails with infection rate indicated that Oncomelonia, indoplanorbis and Bullinus are the predominant genera with infection rate of 31.79, 17.10 and 14.46 percent respectively. However the highest number of the snails collected belonging to the genera Indoplanorbis. At Sheikhupura district, genus wide prevalence of snail indicated that Bullinus, Lymnaea, Indoplanorbis and Physa are the four prominent snails with infection rate of 24.74, 20.57, 14.66 and 13.84 percent respectively. At Sargodha district, genus wise prevalence of snails indicated that Lymnaea, Indoplanorbis, Bullinus and Physa are the four prominent snails with infection rate of 25.09, 14.29, 14.28 and 16.77 percent respectively. At Lahore district, genus wise prevalence of snails indicated that Bullinus Lymnaea, Physa and Indoplanorbis are the four prominent snails with infection rate of 23.37, 18.96, 13.97 and 12.70 percent respectively.While the prevalence at the snail level the Chi square value is 242.944 and the P-Value is 0.0000 1 which is highly significant.
Part 3: deals with the meteorological data ie, temperature, humidity, rainfall and pan evaporation with prevalence of snails and parasites. The temperature and rain fall play very important role in the spread of disease. The ideal temperature ranges form 22-25 °C where development within snail takes place in an efficient manner similarly humidity
f ranges from 55-70% is ideal for the development of the snail and the parasite. Rainfall is very important for the spread of the disease. There is a positive correlation of disease incidence to maximum and minimum temperature, humidity, and rainfall and pan evaporation. It was seen that during summer and autumn, optimum temperature, relative
humidity and rainfall play an important role for rapid propagation of the parasitic life
Part 4: deals with therapeutic trials against Schistosomiasis in buffaloes. A total of 150 animals (140 infected and 10 animals, normal) age ranged 5-9 years and of both sexes naturally infected with Schistosorniasis were used in thirteen controlled experiments. The efficacy of certain indigenous drugs, including Nigella sativa (Kalongi) , Caesalpinia
Crista (Karangwa), Lagenaria siceraria seeds (Kadoo ke Beej), Sausseria lappa (Qushte-e-Shreen) and Praziquanlel was compared with each other and control. Efficacy was quantified by determining the difference of egg per gram faeces (EPG) pre and post treatment. After the single dose of 50, 75 and 100 mg 1kg body weight of Nigella sativa (Kalongi) reduced EPG by 65.85, 68.29 and 71.79 per cent, respectively. After the second dose the respective reduction in EPG was 85.36, 92.68 and 94.87 percent. Caesalpenia crista at three dosage levels i.e.50, 75 and 100 mgI kg body weight caused 46.34, 53.65 and 59.52 percent reduction respectively while the reduction in EPO after second dose was 82.92, 90.24 and 92.85 percent respectively. Lagenaria siceraria Seed at three dosage levels i. e., 50, 100 and 150 mg/ kg body weight caused the reduction in EPG reduction 47.61, 52.63 and 64.10 percent respectively, while after second dose, counts as the reduction 80.95, 86.84 and 92.30 percent respectively. Sausseria lappa at three dosage levels i.e., 100, 150 and 200 mg/ kg body caused EPG reduction as under 50.00, 53.48 and 56.09 percent respectively, while after second dose the reduction in EPG count was 71.42, 81.39 and 85.36 percent respectively. Where as Praziquantel at the dose of 10 mg/body weight caused reduction in EPO 66.66% while after the second dose the reduction in EPG count was 97.43 percent. The efficacy order was Praziquantel, Nigella saliva, Caesalpinia crista, Lagenaria siceraria and Sausseria lappa. No side effects with any drug were noted. All animals showed clinical improvement after the treatment.
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Studies On Growth Performance, Morphology, Reproductive Traits And Behavioral Aspects Of Ring Necked Pheasants In Captivity
Material type: Book ; Literary form:
Publisher: 2015 Dissertation note: Besides ecological importance, pheasants also have aesthetic values which mainly contribute to their decline in population due to poaching in their native habitat. Among 49 species of pheasants in the world, 5 are endemic to Pakistan with distribution in the Himalaya and remote northern parts of the country. Due to increase in human population, intrusion, poaching, and habitat disturbance pheasants are threatened. However, much of the work on captivity is based on production rather morphological, ornamentation or reproductive traits which could have important implications for the management of wild and captive populations as a whole. Some studies still remain to be explored.
Present study was conducted to find out the relationship of egg weight with egg quality parameters and growth traits of ring-necked pheasant Phasianuscolchicus. Total of 450 eggs were collected and were divided into three egg weight categories viz. light (20.0-26.0g), medium (27.0-32.0g) and heavy (33.0-40.0g) egg weight eggs. Fifty eggs for each of the egg category were reserved for the evaluation of internal egg quality parameters. External egg quality parameters i.e. egg length, breadth, egg volume and surface area varied significantly (P<0.05) between all the three egg weight categories. Similarly, significantly higher albumen and yolk weight were recorded in heavy weight egg category while non-significant relationship of egg weight was observed for shell and membrane thickness, yolk percentage, yolk index, yolk pH and albumen pH of the egg. The hatching percentage for the remaining 300 egg kept in incubator was 47.33%. Forty chicks from each of the egg weight category were selected and chick weight, wing length and wingspan were taken at the time of hatching and thereafter increase in these parameters were noted on weekly basis. The effect of egg weight on chick weight, live weight gain, wing length and wingspan was significant (P<0.05) from 1st to 12th month of age. Our studies revealed that egg weight has strong influence on external and internal characteristics of the eggs and the growth parameters in P. colchicus chicks.
The present study was planned to evaluate the time budgets of ring necked pheasants Phasianuscolchicus in captivity. The birds were kept in cages of 5 ft × 5 ft × 3 ft (length × width × height) and were housed in a 20 ft × 20 ft (length × width) well ventilated room at Department of Wildlife and Ecology, Ravi Campus, University of Veterinary and Animal Sciences, Lahore. Captive birds were divided into three categories viz. adult male, adult female and chicks and were placed into separate cages. Behavioral parameters viz. jumping, aggression, preening, feather pecking, walking, standing, sitting, litter pecking, drinking, feeding, body shaking, voice call and feather flapping were assessed for 30 birds from each of the three categories through scan sampling. Statistically significant variations were recorded in behavioral aspects among all the three categories. Male birds spent significantly higher times in aggression (155.26±3.10 sec), preening (74.04±3.05 sec), walking (1370.93±54.45 sec), drinking (74.00±3.18 sec), body shaking (24.92 ±3.11 sec), voice call (20.08±3.17 sec) and feather flapping (15.42±2.73 sec) while female P. colchicus spent significantly higher times in sitting (364.57±3.74 sec). Similarly, the chicks spent significantly higher times in jumping (36.17±2.75 sec), feather pecking (265.19±3.17 sec), standing (1230.13±23.86 sec), litter pecking (234.89±2.97 sec) and feeding (115.44±3.11 sec) as compared to the adult female and male birds.
Fecal and blood samples of ring necked pheasants, Phasianuscolchicus were analyzed to record the parasitic prevalence in these pheasants. A total of 1000 samples, 500 blood and 500 fecal samples were collected from Captive Breeding Facilities for Birds, Department of Wildlife and Ecology, Ravi Campus, University of Veterinary and Animal Sciences, Lahore. Parasitic genera identified from blood samples of P. colchicus include Leukocytozoon, Plasmodium and Haemoproteus. Prevalence of Leukocytozoon was 16 % while the prevalence of Haemoproteus was 14.3%. Parasitic genera identified from fecal samples of P. colchicus include Eimeria, Isospora, Trichomonas and Giardia. Eggs of five species of nematodes viz. Capillaria, Syngamus trachea andAscaridia,Heterakisisoloncheand Heterakisgallinarum were also identified from the fecal samples. The ectoparasites include one species of burrowing mite Knemidocoptesmutansand two species of chewing lice i.e. Amyrsideaperdicis and Lipeurusmaculosus.
Variations in hematological parameters during different life history stages were recorded in ring-necked pheasants (Phasianuscolchicus) for a period of 1 year. Thirty birds were selected for analysis of selected hematological parameters viz. red blood cells count, white blood cells count, hemoglobin, mean corpuscular hemoglobin concentrations, total serum protein and leucocyte count. These birds were kept in cages, each cage having separate drinking and feeding facilities. Five birds per cage were confined and these cages were housed in a well-ventilated 20 × 20 feet (length × width) room. Blood samples were taken from ulnar vein and variations in blood parameters were recorded on monthly basis. Significantly, lower RBC’s count was observed during 2nd month of age while during same month significantly higher WBC count was noticed. Significantly lower values of hemoglobin were observed during 1st and 2nd month of age. Significantly higher mean corpuscular hemoglobin concentration percentage (MCHC%) was recorded during 6th and 7th month of age. Lower packed cell volume (PCV) values were observed during 2nd and 6th month of age while the total serum protein concentrations were recorded maximum during 10th month of age. Significantly, higher heterophils count was recorded during 2nd and 10th month of age while maximum lymphocyte count was observed during 2nd and 7th months of age. Significantly, higher concentrations of monocytes were recorded during 11th, 12thand 10th month of age. The eiosinophils count varied from minimum (110 ± 13.50) during 3rd month to maximum (902 ± 93.22) during 11th month of age. Similarly, significantly higher values of basophils were recorded during 1st month of age. It can be concluded from the present study that the blood profile of the pheasants changes with age.
Variations in hematological parameters for adult male and female ring-necked pheasants (Phasianuscolchicus) were recorded. Forty adult ring-necked pheasants (20 ♂, 20 ♀) were kept in separate cages, each cage having separate drinking and feeding facilities. Five birds per cage were confined and these cages were housed in a well-ventilated 20 × 20 feet (length × width) room. Blood samples were taken from ulnar vein. Different blood and serum chemistry parameters such as red blood cells (RBCs), white blood cells (WBCs), hemoglobin (Hb ) concentrations, mean corpuscular hemoglobin concentration (MCHC), packed cell volume (PCV), heterophils, lymphocytes, monocytes, eosinophils, basophils, ALP, uric acid, cholesterol, total serum protein, albumin and creatinine were determined among adult male and female pheasants. Non-significant differences in RBCs, WBCs, heterophils, eosinophils, MCHC and Hb values were observed among male and female pheasants. Significantly, higher values of lymphocytes, monocytes and PCV were observed in males while higher basophil count was observed in female as compared to male birds. Significantly higher values for ALP, cholesterol, total serum protein and creatinine were observed in males while higher uric acid values were observed in females as compared to male P. colchicus. However, non-significant differences in albumin were recorded among male and female birds.
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Mutational Screening Of The RB1 Gene In Pakistani Patients With Retinoblastoma
Material type: Book ; Literary form:
Publisher: 2015 Dissertation note: Retinoblastoma is a neonatal intraocular tumor caused by biallelic inactivation of RB1 gene. Rb
patients and asymptomatic carriers undergo a series of clinical tests for diagnosis and tumor
treatment. These clinical examinations prove to be expensive and time consuming. On the other
hand if the proband’s RB1 gene mutation status is determined by genetic testing, it can prove as
more significant and cost effective diagnostic methods. Secondly, only those asymptomatic or at
risk carriers with the mutation, require clinical surveillance while those proven to be unaffected
do not require additional clinical examinations. Furthermore early diagnosis of Rb by molecular
testing can enable and enhance clinical management, earlier treatment, follow-up care, carrier
screening, genetic counseling, prenatal diagnosis and reproductive planning in predisposed
families. Irrespective of the importance of molecular testing of Rb patients, in Pakistan only a
few clinical reports on Rb are available so, there was a dire need to find RB1 mutations in
Pakistani Rb patients and to set a molecular based diagnosis for poor affected families. Keeping
in view the importance of molecular diagnosis, in this study a reliable genetic test has been
developed to detect the RB1 germline mutations in Pakistani Rb patients.
During this study, 70 Rb patients including 38 unilateral and 32 bilateral cases were enrolled,
from different regions of Pakistan. By using direct sequencing method, seven novel and twelve
reported RBI mutations were found. The novel mutations included three frameshift mutations
(c.1116_1119delCACT in exon 11, c.1436_1437delAC in exon 16 and c.2060_2061insTCATT
in exon 20) and four substitutions (c.148G>T in exon 2, c.610G>T in exon 2, g.94G>C in exon
7, c.947A>T in exon 10 and g.1991G>C in promoter region) while twelve reported mutations in
22 patients included, 9 substitutions (c.160G>T in exon 2, c.289G>T in exon 3, c.751C>T in
exon 8, c.920C>T in exon 9, c.967G>T in exon 10, c.1072C>T in exon 11, c.1654C>T in exon
17, c.2063T>C in exon 20 and c.2359C>T in exon 23), one frameshift mutation (c.772_776del in
exon 8) and two splice site mutations (c.380+1G>T and c.1215+1G>A in intron 3 and 12
respectively). Mutation detection rate was found to be 77.8% in (7/9) bilateral familial, 50% in
(2/4) unilateral familial, 56.5% in (13/23) bilateral sporadic and 14.7% in (5/34) unilateral
sporadic patients while overall rate of mutations in bilateral and unilateral patients was detected
as 62.5% (20/32) and 18.4% (7/38) respectively. Beside mutations one novel c.940-64C>T
(intron 9) and nine reported intronic variants c.380+45 C>T (intron 3), c.501-77G>A (intron 4),
c.1128-72T>G (intron 11), c.1695+99A>T (intron 17), c.1695-1696delAA (intron 17), c.1815-
104A>G (intron 18), c.1961-10T>C (intron 19), c.2663+33T>C (intron 25) and c.2664-10T>A
(intron 25) were also found. Carrier screening facility was also provided to six asymptomatic
siblings (as possible carriers) of familial proband but none of them was found to be diseased.
Hopefully, in future the findings and developed protocol of this study will help to reveal the
molecular basis of Rb in Pakistani Rb patients which additionally help to secure vision and life
of Rb patients. Further, in Pakistan there is dire need to develop “National Rb Registry Centre”,
to register all new Rb cases for finding incidence rate and prevalence of Rb in Pakistan. Beside
this other related issues like financial constraints, health education, planning and awareness
about Rb, occupational training for health providers, capacity building for neonatal
ophthalmologic screening and cosmetic rehabilitation for surviving Rb patients are important and
Availability: Items available for loan: UVAS Library [ Call number: 2370-T] (1).
Biological Studies on Various Avian Influenza Virus Types In Poultry
Material type: Book ; Literary form:
Publisher: 2011 Dissertation note: Theses submitted with blank cd.
Availability: Items available for loan: UVAS Library [ Call number: 2390-T] (1).
Prevalance, Diagnosis and Economic Losses Due to Bovine Cysticercosis in Punjab
Material type: Book ; Literary form:
Publisher: 2014 Dissertation note: Theses submitted with blank cd.
Availability: Items available for loan: UVAS Library [ Call number: 1379-T] (1).
Effects Of Ginkgo Biloba And Panax Ginseng On Metabolism Of Carbohydrate, Lipids And Insulin Receptor Genes In Diabetic Rats
Material type: Book ; Literary form:
Publisher: 2014 Dissertation note: Diabetes is a major public health issue. As conventional pharmaceutical agents have greater incidences of adverse effects so the interest in the natural remedies has increased greatly in the last few decades. Ginkgo biloba leaf extract (GBE) and Panax ginseng root extract (PGE) are ancient Chinese herbal drugs that have prominent position in the list of the best-selling natural remedies and are increasingly being used for the treatment of diabetes. The anti-diabetic effect of GBE is attributed to flavonoides while that of PGE is attributed to ginsenosides. In this study, GBE and PGE in combination showed significantly higher anti-diabetic effects than individual extracts in diabetic rats.
Adult Wistar rats were allowed to feed on a high fat diet (HFD: 12.7% maize starch, 6.5% dextrose, 3.9% sunflower oil, 31.3% beef tallow and 28.6% casein by weight) for two weeks. The rats were divided into seven groups (08 rats in each group): Non-diabetic control group, Diabetic group, Diabetic + 100 mg/kg G. biloba leaf extract treated group (GBE), Diabetic + 300 mg/kg, P. ginseng root extract treated group (PGE), mixed 1 group : Diabetic + combination of both GBE and PGE at dose of 200 mg/kg/day (50mg/kg/day of GBE and 150mg/kg/day of PGE), mixed 2 group : Diabetic + combination of both GBE and PGE at dose of 400mg/kg/day (100mg/kg/day of GBE and 300mg/kg/day of PGE), mixed 3 group : Diabetic + combination of both GBE and PGE at dose of 600mg/kg/day (150mg/kg/day of GBE and 450mg/kg/day of PGE). At the end of the 14th day, the rats were kept in fasting condition overnight and then a single intra-peritoneal injection of alloxan monohydrate (Sigma, USA) dissolved in 0.5 ml of saline solution at a dose of 120-130 mg/Kg body weight was injected in all rats except for the non-diabetic group which were injected with an equal volume of normal
saline. Body weight (BW) and blood glucose were measured at week 1 and week 14. At the end of the experimental period, blood samples in fasting/ basal state were collected from heart puncture for the biochemical parameters. Liver, muscles and adipose tissue were also collected for mRNA expression of genes involved in carbohydrate and fat metabolism.
Results were expressed as Means ± S.E.M. Statistical analyses was performed using Statview software (SAS Institute Inc., SAS Campus Drive, Cary, NC, USA). Two-ways repeated measure ANOVA followed by PLSD Fisher's test was performed for BW and blood glucose to assess the effects of time and herbal drugs. For the rest of the parameters, one-way ANOVA followed by PLSD Fisher's test was performed to assess the effect of herbal drugs. Differences were considered significant at P < 0.05.
A significant (P < 0.0001) reduction in the BW of the diabetic group was recorded compared to non-diabetic rats and a significant reduction in BW was observed after treatment in all the five treated groups compared to diabetic group. Glycemia was significantly higher in the diabetic rats (P < 0.0001) compared to non-diabetic rats and a significant reduction in the blood glucose level was recorded in all the five treated groups (P < 0.0001) group in comparison to the diabetic group. A significant reduction for fasting serum glucose (FSG) (P < 0.0001) was recorded for all the five treated groups compared to the non-treated diabetic rats. We linked the reduction in hyperglycemia to the mRNA expression of genes involved in glucose metabolism. In particular, we studied the gene expressions of GLUT-4, insulin receptor (IR), insulin receptor substrate-1 (IRS-1) and phosphoenolpyrovate carboxykinase (PEPCK) in liver, muscle and adipose tissue. A significant up-regulation for the mRNA expression of GLUT-4 was observed only in muscle in all the five treated groups, i.e. GBE (P < 0.001), PGE (P < 0.001), mixed 1 (P < 0.0001), mixed 2 (P < 0.0001) and mixed 3 (P < 0.0001). We found a significant down-
regulation in the mRNA expression of IR in muscle (P < 0.0001) and adipose tissue (P < 0.05) in the diabetic group compared to non-diabetic rats, however, a significant up-regulation was found in mixed 3 group in muscle (P < 0.001) and adipose tissue (P < 0.05). We found a significant down-regulation (P < 0.001) for IRS-1 in liver in diabetic state and a significant up-regulation was recorded in GBE (P < 0.05) and mixed 3 (P < 0.05) groups only. We found a significant down-regulation of IRS-1 in muscle (P < 0.0001) and adipose tissues (P < 0.0001) in the diabetic group. None of the treated group showed significant results in muscles however, a significant up-regulation was found only in PGE (P < 0.001) and in the mixed 3 group (P < 0.0001) in adipose tissue. A significant up-regulation was recorded for PEPCK in GBE (P < 0.05), mixed 1 (P < 0.05), mixed 2 (P < 0.05) and mixed 3 (P < 0.05) groups in liver.
A significant increase of blood cholesterol was found in rats in the diabetic state (P < 0.0001) and a significant reduction was found only in the mixed 3 (P < 0.001) treated group. A significant decrease was found for VLDL-C in mixed 1 (P < 0.05), mixed 2 (P < 0.0001) and mixed 3 (P < 0.0001) groups. A significant decreased was observed for LDL-C in mixed 1, mixed 2 and mixed 3 (P < 0.0001) groups which previously found to be enhanced in diabetic condition. In case of HDL-c a significant decreased was found for GBE (P < 0.001), PGE (P < 0.05), mixed 1 (P < 0.001), mixed 2 (P < 0.0001) and mixed 3 (P < 0.0001) which was previously found to be increased in the diabetic group (P < 0.0001). Conversely, a significant increase was seen for TG (P < 0.0001) in the diabetic state and a significant reduction was found in all the five treated groups (P < 0.0001). We further studied genes involved in lipid metabolism. A significant up-regulation was found for SREBP-1c in diabetic group (P < 0.0001) and a significant down-regulation was found to occur in mixed 2 (P < 0.05) and mixed 3 (P < 0.001) treated groups compared to untreated diabetic rats. In the liver, a significant up-regulation
in the mRNA expression of FAS was found only in mixed 2 (P < 0.05) and mixed 3 (P < 0.05) treated groups which found to be down regulated in the untreated diabetic group (P < 0.001). A significant down-regulation in the mRNA expression of PPAR-α was found in diabetic rats skeletal muscle (P < 0.05), however, a significant up-regulation was found in GBE (P < 0.001), PGE (P < 0.05) mixed 1 (P < 0.001), mixed 2 (P < 0.001) and mixed 3 (P < 0.001) treatment groups in comparison to diabetic rats. We studied PPAR-γ in adipose tissue and found a significant up-regulation in PGE (P < 0.05), mixed 1 (P < 0.001), mixed 2 (P < 0.001) and mixed 3 (P < 0.0001) groups which had previously been found to be down regulated (P < 0.001) in diabetic rats compared to non-diabetic rats.
We found that the body of the diabetic rats suffer with oxidative stress and measured a significant decrease for CAT (P < 0.0001) in diabetic group and significant increase was found in GBE (P < 0.05), PGE (P < 0.05), mixed 1 (P < 0.05), mixed 2 (P < 0.05), mixed 3(P < 0.05) groups compared to diabetic rats. Whereas, a significant decreased was recorded for MDA in GBE (P < 0.05), PGE (P < 0.05), mixed 1 (P < 0.001), mixed 2 (P < 0.001) and mixed 3 (P < 0.0001) groups, which previously showed a significant increased (P < 0.001) in diabetic group compared to non-diabetic. We linked oxidative stress with TNF- α and found a significant up-regulation (P < 0.0001) for all the three studied organs in diabetic groups compared to the non-diabetic group. In case of liver a significant down-regulation was found for GBE (P < 0.0001), PGE (P < 0.0001) and mixed 3 (P < 0.0001) groups compared to untreated diabetic rats. A significant down-regulation in the expression of TNF- α in muscle was recorded only in the mixed 2 (P < 0.001) and mixed 3 (P < 0.0001) groups compared to diabetic rats. However, a significant down-regulation in the expression of TNF- α in adipose tissue was observed for all the treated groups (P < 0.0001 for all groups) in comparason to the diabetic group.
For serum creatinine a significant enhancement was observed for PGE (P < 0.05), mixed 1 (P < 0.05) and mixed 3 (P < 0.05) groups which were previously found to be reduced in diabetic rats. A significant increase for AST was found in diabetes (P < 0.0001) compared to non-diabetic rats, while a significant reduction was found to occur only for PGE (P < 0.05), mixed 2 (P < 0.05) and mixed 3 (P < 0.001) treated groups in comparison to the untreated diabetic group. Like AST a significant reduction was recorded for ALT in the diabetic group (P < 0.001) and only GBE (P < 0.001), PGE (P < 0.05) and mixed 3 (P < 0.05) showed a significant decreased in ALT level compared to untreated diabetic rats.
In conclusion, we found that both GBE and PGE have strong individual anti-hyperglycemic, anti-hyper-triglyceridemic and anti-oxidative effects in an alloxan monohydrate induced rat model of diabetes. Both also showed strong influence on the activation on the expression of genes involved in the metabolic pathways of glucose and lipid which previously became dysfunctional in diabetic rats. When both these natural remedies were given in combination, synergistic effects were recorded in a dose dependent manner. Further work is needed to evaluate the way by which human beings suffering from diabetes are safely treated with these herbal remedies.
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Serodiagnosis And Chemotherapy Of Trypanasomiasis In Camels In Balochistan
Material type: Book ; Literary form:
Publisher: 2015 Dissertation note: Trypanosomiasis (Surra or Rotten disease) is a disease of animals such as cattle, buffalo,
horses and camels caused by protozoan parasite of mammalian blood. In camels, the clinical
manifestations of trypanosomiasis by T. evansi can be acute or chronic. The rypanosomiasis
symptoms include fever, emaciation, anemia, weight loss, edema, lymphadenomegaly,
conjunctivitis and occasionally sudden death of animal. However, the diagnosis of disease
always remains a challenge for veterinarians and animal healthcare professionals, since the
parasitemia is often very low in a majority of infections.
Keeping in view the importance of Trypanosomiasis, the present study was designed to
evaluate the present status of Trypanosomiasis in Province Balochistan of Pakistan. The two
districts Musa Khail and Jhal Magsi with thick populations of camels and have different climate
and geographical distribution were selected. The questionnaire was developed for survey of
Camel farmers and information regarding the age of respondent, experience, type of
community, feeding/watering pattern of camels, prevailing camel diseases in the area, treatment
facilities, traditional remedies used by them against various diseases in camels and economic
losses were collected. A total of one thousands and forty (n=1040) camel owners/respondents
from three groups viz settled, transhumants and nomads were interviewed in Districts, Musa
khail and Jhal Magsi during the year 2011.
In first experiment, a total of 1600 blood samples were randomly collected from camel
population of both districts i.e. 800 camels from each districts. Camels were sub-grouped
according to sex (male and female) and age (upto 2 years, 2-3 years and above 4 years). Blood
samples were collected from each animal and processed for blood smears examination and PCR
for confirmation of Trypanosomiasis. The blood samples were also processed for hematological
studies to evaluate the effect of Trypanosomiasis on different blood parameters. The blood smear
examination showed prevalence of 11.87 percent (95 out of 800 samples) in District Musa Khail
and 17.12 percent (137 out of 800 samples) in district Jhal Magsi, indicating higher prevalence in
hot climate when compared with cold climate. However, the higher prevalence in females,
animals aged above 4 years and during summer season in both districts.
In second experiment, the prevalence of trypanosomiasis was recorded through PCR
amplification in 200 samples (100 from each Districts Musa Khail and Jhal Magsi). The
prevalence was found to be 25 percent in District Musa Khail and 39 percent in District Jhal
Magsi. PCR test proved to be most sensitive and specific technique for diagnosis and
confirmation of Trypanosomiasis. The hematological tests including hemogram, lecucogram,
serum protein, and serum electrolyte and serum enzyme showed fluctuation from its normal
ranges. Moreover, hemogram (RBC, PCV and Hb) showed significant decrease than the normal
values while in Leucogram showed significant increase than normal values.
In third experiment chemotherapy trials on 25 camels divided in 5 groups A, B, C, D and E were
conducted. Single dose of anti-trypanosome drug (Samorin) Isometamidium chloride @ dose
0.5mg/kg deep I/M to group A. Dried leaves of Azadirachtaindica (Neem) @ 60 gm/animal, to
group B. Dried and grind fruits of CitrilisColocynthis (L) Schard @ 50gm/animal, to group C.
Group D was control diseased and Group E was control healthy group. Blood samples were
collected post-treatment at 2nd, 7th and 12th day and the changes in blood picture were analyzed.
The efficacy of drug and plant extract was tested after 72 hours of treatment. Blood samples
were collected from group A, B and C and processed for amplification of PCR. Results of
allopathic showed 100 percent efficacy while result of plant Azadirachta indica (Neem) showed
60 percent t efficacy by amplification of PCR. In group C plant Citrullus Collocynthis showed no
In third experiment, the direct as well as indirect economic losses due to camel Trypanosomiasis
based on the prevalence of Trypanosomiasis, mortality rate, abortion and perceptions of the
respondents were recorded. The camel dies due to Trypanosomiasis in direct visible losses and
invisible losses include reduced fertility, meat loss, low quality of hide, loss of draught power
and traction force and change in herd. While, indirect losses include additional costs of drugs,
veterinarian fee, preventive medicine and quarantine.
The respondents were grouped according to type of communities as Nomads spend
summer in Afghanistan (uplands) and back to Pakistan (lowland) in winter. Hence, their camels
harbor Trypanosomiasis from one border to another border and present trans-boundary impact of
the disease. The present study demonstrates that the respondents above the age of 50 years were
more experienced in disease diagnosis and use of traditional veterinary practices. The settled and
transhumants communities had easy access to government hospitals and private veterinary
clinics while nomads mostly rely on the use of traditional veterinary practices. The most
common prevailing diseases of camel according to respondents were pneumonia, indigestion,
parasitic infestation, mange, lameness, Trypanosomiasis, vector fly and nervous disorders. The
most common clinical signs of camel Trypanosomiasis hyperthermia, anemia, depression,
dullness, emaciation, edema (in dependent parts of body), abortion, nervous signs, circling
movements, trembling, unusual aggressiveness and aimless running were recorded.
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Clinico-Epidemiological And Therapeutic Study On Babesiosis In Different Breeds Of Cattle In Balochistan
Material type: Book ; Literary form:
Publisher: 2015 Dissertation note: Babesiosis which is also called as piroplasmosisis, Texas fever, redwater or tick fever, is an emerging, tick-transmitted (by a vector ixodidea) disease caused by intraerythrocytic parasites of the genus babesia having considerable worldwide economic, medical, and veterinary impact. Keeping in view the importance of babesiosis under local conditions, the present study was designed to evaluate the status babesiosis in Balochistan. For this purpose field and experimental studies were carried in two districts Quetta and Sibi of Balochistan Province to find out the status of babesiosis in Bhag Nari, Holstein Friesian and Crossbred cattle. During field study epidemiological status of babesiosis was highlighted by selecting 600 cattle randomly from each district. The animals were distributed into 2 major groups i.e. Young animals less than 12 months and adult over 12 months of age. These groups were further sub-divided into Young animals (less than 6 months, up to 9 months and up to 12 months) while Adults animals (up to 2 years, 3 years and over 3 years). The vector of babesia was also kept under keen observation for the prevalence/infestation rate, identification and economic losses caused during the course of study. Blood samples were collected from each animal and processed for blood smears examination and PCR for further confirmation of babesia infection. The blood samples were also processed for hematological study to evaluate the effect of babesiosis on different blood parameters. For experimental study 148 animals were selected through clinical signs of babesiosis, blood smear examination and PCR. Out of theses 40 animals were maintained for therapeutic trail to find out the cheapest and easily available drug against bovine babesiosis. For this purpose Neem leaves were used in decoction form while Imidocarb dipopionate was kept as standard control. The
results of epidemiological study revealed higher prevalence of babesiosis (20.5%) in district Quetta while 15.16% was recorded in District Sibi. Similarly higher prevalence was recorded in Holstein Friesian than in Crossbred and Bhag Nari cattle respectively in both districts Quetta and Sibi. Furthermore higher prevalence of babesiosis was recorded in adult groups of Holstein Friesian than in Crossbred and Bhag Nari cattle. Similarly season wise higher prevalence of babesia infection was noticed in summer followed by spring, autumn and winter respectively while higher prevalence was noted in female group of animals than male animals. Blood smears examination and PCR confirmed two babesia species i.e. babesia bigemina and babesia bovis. Similarly Boophilus tick species were identified as the vector of babesia parasites. During present study mixed hemoprotozaon infection of babesia mixed with theileria was recorded in both districts. The results of conventional method and modern diagnostic technique (PCR) revealed that PCR identified higher babesia infection during the entire 4 seasons as well as in all age groups whereas blood smears examination was capable to diagnose babesiosis in adult groups during the months of summer and spring season. Breed wise prevalence was also higher in samples treated with PCR than blood smears examination and even samples that were declared negative by blood smears examination were also found positive. The results of complete blood cell count from blood samples of infected experimental animal showed regenerative, macrocytic hypochromic anemia. Blood smear examination showed presence of many babesia with reticulocytes. Abnormalities in erythrocyte structure were seen. The result of blood parameters of total erythrocyte count, total leukocyte count, packed cell volume and hemoglobin showed significant decrease in all three affected Bhag Nari, Holstein Friesian and Cross bred cattle. The values of MCV and MCH were increased and MCHC was slightly less than normal value. No efficacy of neem decoction was noted against bovine babesiosis.
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Proteomic And Genomic Analysis Of Methicillin-Resistant Staphylococcus Aureus And Efficacy Of Indigenous Medicinal Plants Essential Oils
Material type: Book ; Literary form:
Publisher: 2015 Dissertation note: A Cohort study (prospective and observational) was performed to study the prevalence of Methicillin resistant Staphylococcus aureus from the healthy individuals of community, hospitalized patients and associated health-care workers and indigenous plants essential oils were screened as new, improved & potent antibacterial/s against resistant strains of MRSA.
The method involved isolation and identification of MRSA from surgical wounds of hospitalized patients & associated health care workers in a tertiary care hospital in Lahore and healthy volunteers from the community. Plant essentials oils & extracts were evaluated for their antibacterial activity against selected MRSA isolates. Oils were recovered by steam distillation using an all-glass distillation assembly. Then in vitro sensitivity and MICs of plant essential oils were determined using vancomycin and linezolid as commercial standards. The essential oils were screened further for the active constituents by column chromatography using various solvents and identification of compounds were performed by GC/MS analysis and the fractions which showed prompt results were evaluated for antimicrobial activity against the MRSA isolates in quest to find new therapeutic options. Finally effective essential oils and their active fractions were studied for their toxicity using in vitro Genotoxic assays such as Ames and Comet assays. To further ensure their beneficial effects antimutagenic effect of the essential oils were also studied.
Prevalence of S. aureus among patients was 52.9%, in HCWs 86.5% and in community 74% with an overall percentage of 72.6%. Among S. aureus those declared as MRSA were 91.8% from patients, 50.6% from HCWs and 59.5% from community with an overall percentage of 62.2% MRSA. Among the isolated MRSA overall 90.6% were Coagulase positive and 75.2% were biofilm positive.
The pattern of MRSA resistance against current antibiotics have shown an overall increase in the resistance with maximum shown for lincomycin followed by tetracycline, ampicillin, fusidic acid, amoxicillin and piperacillin with tazobactam. The most effective options among current regime were tigecyclin, amikacin and meropenem showing an overall least resistance. Resistance against linezolid was observed with an overall percentage of 25.6 % and vancomycin 33.3% by disc diffusion method.
The MRSA isolates resistant to one or more groups of antibiotics were declared as MDRs. Among patients and health-care workers all were declared as MDRs where as in community 93.1% isolates were MDRs.
Upon Protein profiling using whole cell proteins 44 bands of the polypeptides were produced with molecular size 10-200kDa from the three sampling groups and were categorized into 5 clusters showing an overall significance correlation with each other explaining an interesting fact that all these strains were interlinked establishing the fact of flow of hospital acquired MRSA in the community and vice versa. This analysis also gave an insight in explaining the fact of horizontal transmission of infection within the hospital.
Keeping in view the raise in resistance among current available antibiotics indigenous medicinal plants essential oils were screened for active constituents exhibiting anti-bacterial effects against MRSA isolates.
Maximum yield was obtained from Carum copticum followed by Cuminum cyminum and minimum yield was obtained in case of Zingiber officinale. Upon qualitative analysis of all five essential oils Carum copticum essential oil showed zones of inhibition greater than the standards vancomycin and linezolid followed Cuminum cyminum and Zingiber officinale in all three
sampling groups. Anethum sowa and Myristica fragrans essential oils showed no activity against MRSA.
Minimum inhibitory concentration of the three essential oils determined by micro broth dilution method indicated that Carum copticum showed least value in all three types of MRSA isolates followed by Zingiber officinale and Cuminum cyminum.
Effective essential oils were further fractioned using silica gel gravity columns. All the fractions obtained were screened for the anti-bacterial activity against all three types of MRSA isolates. Only fraction F1 of Carum copticum showed activity greater than pure essential oil and the two commercial standards of vancomycin and linezolid.
For the identification of active constituents GC/MS analysis was performed on all three essential oils and their respective fractions. In case of fraction F1 the most dominant constituents were Carvacrol, p-Cymene, Ʈ-Terpinene and Apiol. In other two plants none of the fractions were effective. Therefore it was concluded to use pure essential oils in case of Zingiber officinale and Cuminum cyminum rather than their individual fractions and incase of Carum copticum Fraction F1 has shown superior activity.
Finally these essential oils were tested for possible mutagenic effect using bacterial reversion mutation assay and Comet assay. No mutagenic effects were observed at MIC and above doses. These effective essential oils were also evaluated for possible antimutagenic effect. Both Carum copticum and Zingiber officinale essential oils showed strong antimutagenic effects and weak antimutagenic effect by Cuminum cyminum. Upon analysis of nuclear damage none of the plants essential oils and fraction F1 of Carum copticum showed genotoxic effects and indicated to be safe. Thus from the study it was concluded that Carum copticum essential oil and its fraction F1 were the most effective to be further investigated as an alternative treatment for MRSA infections.
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Assessment Of Avian And Mammalian Diversity At Selected Sites Along River Chenab
Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: The River Chenab is an important wetland of Punjab province and the tree plantations around the
river are the part of tropical thorn forest. But as a consequence of deforestation much of the
natural forested areas have been turned to agricultural land. The main objective of study was to
assess the avian and mammalian diversity of the study area; to identify and assess anthropogenic
impacts on avian and mammalian diversity of the study area; and to explore the level of humanwildlife
conflict selected sites of river Chenab i.e. district Sialkot, district Gujrat and district
Gujranwala from May, 2013 through April. Surveys were made during dawn (5:00 am to 8:00
am) and dusk (4:00 pm to 7:00 pm).
During the waterfowl study recorded 51 species belonging to 33 genera, 16 families and 8 orders
were recorded from the study area. Throughout the year a total of 2531 birds from recorded from
head Marala, 2026 from the head Khanki and 2230 from head Qadirabad. Diversity indices were
analyzed through statistical software PAST version 2.17 C. The Shannon-Weiner diversity index
at head Marala was 2.62, at head Khanki it was 2.64 while at head Qadirabad it was 2.78. It can
be concluded from the present study that the River Chenab is waterfowl rich and should be
declared as protected site for waterfowls.
The study area was divided into different habitat types on the basis of vegetation and
urbanization and was designated as forest habitat (FH), wetland habitat (WLH), rural forest
habitat (RFH), agriculture habitat (AH), agriculture rural habitat (ARH), urban non vegetative
habitat (UNVH) and urban vegetative habitat (UVH). A linear count method was applied and
data was collected through direct and indirect observations. Habitat preference of the birds varied
f declined from forested habitats to the urban landscapes. It can be concluded from the study that
many of the avian species are habitat specific and the connection/corridors between similar
habitat types might be fruitful for the conservation of avian species.
The anthropogenic impacts and habitat preferences of mammalian species along river Chenab,
Pakistan was also assessed the mammalian diversity was recorded along forested landscapes,
cultivated plantations, semi-urban and urban areas. The data on diversity and distribution of
various mammalian species was collected through point count method viz. direct observation
(personal count and record voices) and indirect observation (presences of carcasses, fecal pellet,
pug marks and meeting with local communities). The habitat preferences of large, medium and
small mammals varied significantly. A decline in mammalian diversity was observed from forest
habitat to urban landscapes. Indian wild boar, Asiatic jackal, Indian fox, jungle cat, Indian
pangolin and long eared desert hedgehog preferred forested areas as well as slightly modified
habitats while Northern palm squirrel, house mouse, house shrew and rat species preferred
human habitations. Similarly, few species such as the small Indian mongoose, Soft-furred field
rat, short tailed mole rat, Asiatic jackal and Indian gerbil preferred cultivated areas. It can be
concluded that many of the mammalian species are habitat specific and corridors and
connections between different landscapes are important for the conservation of mammalian
Medicinal and cultural significance of avian species along the River Chenab were assessed
through Relative Popularity Level (RPL) and Rank Order Priority (ROP). One hundred and nine
persons were interviewed and data regarding socio-economic status of the respondents,
qualitative data on cultural significance from three selected districts. The compiled data are
analyzed using different quantitative tools, such as relative frequency of mention (RFM),
fidelity level (FL), relative popularity level RPL and rank order priority (ROP). Out of total 155
avian species recorded from the study area, 28 have medical importance while local people were
using feathers of almost all the bird species for making different toys. Ten species were most
popular and highest RFM values (0.58) were recorded for house sparrow (Passer domesticus).
Similarly, highest FL values (100%) were recorded for house sparrow (P. domesticus) and
domestic chicken (Gallus gallus). These studies indicated that the area is rich in avian diversity
and many of these species have medical and cultural significance for the locales.
Mammals are source of food and medication for humans from ancient times. A survey was
conducted along the Rver Chenab, Punjab, Pakistan and 109 persons were interviewed to
investigate the extent of human dependency on mammalian species of the area. A total of 30
mammalian species were recorded from the study area. Highest relatively frequency of mention
(RFM) values (0.5) were observed for desert hare, Lepus migricollis dayanus while maximum
(100%) fidelity level (FL) was recorded for cow Bos gaurus, sheep Ovis aries and cat Felis
domesticus. Seven species were most popular. It can be concluded from present survey that local
people have strong association with mammalian species of the study area and dependent for
food and medicines on these species. In depth studies are recommended to explore medicinal
importance of the species.
The study area was part of tropical thorn forest but a larger portion has been changed into
agricultural land or human habitations. Data regarding socio-economic value of area, financial
losses to crops and livestock, peoples’ attitude and tolerance towards wildlife, protection
methods for livestock and crops from predators and profile of 150 respondents were collected
through a questionnaire. The age of the respondents was between 20 to 65 years, out of them
54% were literate, 99% were Muslims and all these respondents were from different professions
viz. farmers (32%), livestock managers (37%) and others (31%). Most of the respondents (52%)
were unaware about the role of wild species in ecosystem, certain respondents (28%) disliked
wild species in their areas and 20% respondents had positive view about wildlife in the area. The
collected data revealed that crops are mostly damaged by the Indian wild boar (42%), Asiatic
jackal (34%), diseases (11%), Indian crested porcupine (6%) and others (7%) including rats,
squirrels, crows and sparrows. Similarly, the livestock animals are affected mostly by diseases
(36%), Asiatic jackal (34%), jungle cat (10%), Indian fox and others (6%) including raptor birds.
Most of the respondents were of the view that wildlife is declining in the area.
The River Chenab is an important wetland of Punjab, Pakistan. Water of the river is becoming
pollutedt due to anthropogenic impact i.e. industrial waste, urbanization, agriculture
intensification. The main objectives of the study were to know the diversity and distribution of
fish species of river Chenab. Both, direct and indirect methods were applied to find out fish
diversity of the area. The diversity indices were analyzed through statistical software PAST
version 2.17 C. During the sampling 34 species was recorded from the river Chenab. The
diversity indices indicate that higher diversity is present at the head Qadirabad than head Khanki
and Marala. The reason is that there is present large number of natural and manmade ponds;
during the flood these pond fishes move to the river further eggs and fingerlings move to rivers
through birds and fisherman.
Availability: Items available for loan: UVAS Library [ Call number: 2520-T] (1).
Studies on Pathogenesis and Molecular Diagnosis of Infectious Bursal Disease Virus In Broiler Chicken
Material type: Book ; Literary form:
Publisher: 2015 Dissertation note: Blank CD.
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Welfare Assessment Of Wild Turkeys (Meleagris Gallopavo) Subjected To Free Range, Semiintensive And Confinement Rearing Systems
Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: The volume of turkey production is as small compared to the broiler production, although, the
turkey industry has attained a significant increase since 1980, rising from 122 million to 226
million turkeys by 2006 within the European Union, while values of turkeys produced in the US
during 2010 was $4.37 billion. The s scientific literature about welfare of confinedly reared
turkeys is scarce compared to other poultry birds. There is a need for more insight into the
factors that enhance turkey welfare, not only to sustainable production of turkey meat where the
public demands s management practices for the welfare of turkeys, but also because the
information is needed to decrease losses due to poor performance of the birds.
Besides ecological importance, Meleagris gallopavo also have aesthetic values which
mainly contribute to decline in population of turkeys due to poching in their native habitat.
Demand for poultry meat is increasing throughout the world, due to of its lower cost when
compared to other meats such as pork and beef, and also lower fat content and excellent source
of protein. The consumption of turkey meat is about 4.7 million metric tons annually throughout
of the world and with the US accounting for 50% of all consumption.
A total of 120, day-old turkey (Maleagris gallopavo) chicks were arranged into four
groups. Four diets having varying levels of crude protein (CP) viz. 16% CP, 18% CP, 20% CP
and 22% CP were prepared and were designated as control (T0), treatment 1 (T1), treatment 2
(T2) and treatment 3 (T3) diets, respectively. Group 1 birds were fed with T0 diets, group 2 with
T1 diets, group 3 with T2 and group 4 birds were fed with T3 diets. Significantly higher weight
gain 2950.86±1952.58g, body length 30.064±11cm, beak length 3.53cm, body girth
43.41±16.30cm, thigh length 18.18±6.47cm, sternum-length 11.98±4.70cm, wing-length
47.10±16.17cm, wingspan 101.36±34.32cm, shank-length and tarsus-length 13.78±4.47cm were
recorded in M. gallopavo fed with T3 diets. Feed conversion ratios (FCR) for T0, T1, T2 and T3
diets were 2.296, 2.236, 2.099 and 1.934, respectively. Internal and external quality parameters
of egg were also analyzed.
The time budget of turkeys (Meleagris gallopavo) reared under free-range and
confinement rearing systems was recorded and compared from day old chick to the sixth month
of age. Throughout the study period, M. gallopavo reared under free-range rearing system spent
relatively greater time litter pecking (23.51%) followed by walking (19.99%), feeding (16.33%),
preening (13.72%), feather pecking (6.07%), aggression (5.94%), drinking (5.90%), immobility
(2.36%), standing (2.29%) and jumping (1.96%). Similarly, the birds reared under confinement
rearing system spent relatively greater time in lying (17.82%) followed by litter pecking (15.71),
preening (12.93%), walking (11.47%), standing (8.35%), drinking (8.31%), aggression (6.85%),
feeding (6.46%), feather pecking (6.04%), immobility (4.59%) and jumping (1.46%) behavior. It
was observed during the present study that the birds reared under free-range rearing system spent
significantly greater time in litter pecking, walking and feeding behaviors as compared to the
birds reared under confinements. These behaviors are indicators of good health of the animals,
therefore free-range system is recommended over confinement rearing system for farming of M.
Growth performance, carcass yield and meat sensory quality attributes of turkeys
Meleagris gallopavo reared under free-range and confinement rearing systems were compared.
There were two treatments, i.e. indoor and free range, each containing 25 birds. In indoor
treatment, the turkeys were raised in a 20 × 20 feet (length × width) well ventilated room while
in free-range treatment, the birds were housed in an open shed having same dimensions i.e. 20 ×
20 feet (length × width), however, an additional free-range grassy paddock was provided to
them. To compare sensory attributes, 4 ready to-cook turkeys having equal size were selected
from each the rearing systems, were skinned and boneless breast fillets and thighs were prepared,
weighed and steam roasted. The roast breast meat of free-range turkeys was darker and yellower
than thigh meat from indoor birds. Score for breast meat tenderness and appearance varied
significantly (P<0.05) between free-range and confinement reared birds. However, nonsignificant
differences were recorded for thigh meat tenderness between free-range and confined
turkeys. Significantly higher (P<0.05) meat redness was recorded in thigh meat of turkeys reared
in free-range system than thigh meat of confined M. gallopavo while non-significant differences
in breast meat color were observed between free-range and confined birds. Significantly higher
juiciness and oiliness was recorded for confined birds as compared to the free-range reared
Morphological characteristics of turkey Meleagris gallopavo semen were recorded and
hatchability percentages of the artificially inseminated hens were assessed. To study the effect of
natural mating on hatchability 15 (12♀, 3♂) M. gallopavo were randomly selected and both the
genders were kept separately for two weeks, thereafter these birds were arranged in three groups,
each containing one male and four female birds. For AI, male (n = 3) and female (n = 12) birds
were kept separately through the experiment and female birds were inseminated on weekly basis.
Male birds were conditioned and trained for semen collection through abdominal massage and
morphological characteristics of the collected semen were recorded. Mean abnormal
spermatozoa rate was 14.61±1.61% with most common acrosomal defects 36.67±3.80%
followed by defects in mid-piece 8.59±0.24%, head defects 7.15±1.21% and tail defects
6.69±1.97%. Laid eggs were collected, stored and were incubated for hatching. Hatchability
percentage in artificially inseminated hens was greater 88.85% than natural mating 82.35%.
Variations in serum biochemical profile with increase in age were analyzed in turkeys
(Meleagris gallopavo). Gender-wise variations and effect of rearing systems i.e. free range,
semi-intensive and confinements were also assessed from the 1st to 6th month of age. Variations
were recorded in serum biochemical profile with increase in age. Significantly (P<0.05) higher
values of cholesterol 8.39±0.48 mmol/L, globulin 2.71±0.34 g/dL, albumin g/dL 3.82±0.58 g/dL,
total protein 5.35±0.55 g/L, urea 26.95±0.65 mg/dL, creatinine 0.83±0.01 μmol/L, alanine
aminotransferase (ALT) 35.56±1.16 iu/L and aspartate aminotransferase (AST) 44.16±1.83 iu/L
were recorded for adult birds while alkaline phosphatase (ALP) values were significantly higher
104.86±16.39 iu/L in grower birds. Similarly, the rearing systems also influenced biochemical
parameters of M. gallopavo and significantly (P<0.05) higher values for cholesterol 6.18±0.07
mmol/L and ALT 31.26±1.67 iu/L were recorded in birds reared under confinement rearing
system. Values for globulin 1.92±0.36 g/dL and creatinin 0.77±0.08 μmol/L were higher in birds
reared under semi-intensive rearing system while significantly higher urea concentrations
33.45±3.39 mg/dL were recorded from the serum of the birds reared under free range rearing
system. However, non-significant variations in albumin, total protein, ALP and AST were
recorded for all the three rearing systems.
Significantly higher values of cholesterol, globulin, albumin, creatinine and ALT were
recorded from the serum of male M. gallopavo while urea and ALP were higher in hens’ serum.
However, non-significant variations were recorded for total protein and AST among both the
genders. It can be concluded from the present study that age, gender and rearing systems strongly
influence blood biochemical profile and hence the immunity in M. gallopavo.
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Study Of Factors Affecting Quality Of Silage As A Component Of Total Mixed Ration On Growth And Production Performance In Nili-Ravi Buffaloes
Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: Silage production is at initial stages to be a part of animal agriculture in Pakistan. The lack of research on silage making and its benefits for livestock production under local conditions is an important factor for slow propagation of silage in our country. Under such scenario a multi-step study was conducted. At first, the effect of proper maturity stage for harvesting different fodders was investigated, and then the effects of silo type and silage additives were assessed on silage quality. In last part of the study the feeding trials were conducted on growing calves and lactating buffalo to evaluate the effect of silage feeding on growth and milk production respectively.
In all the three fodders i.e. oats, maize and sorghum, the full bloom stage for harvesting produced the best results regarding silage quality and fermentation characteristics. Although the trench silo produced best results regarding fermentation characteristics and silage quality, the expected operational cost and dry matter losses during face management for trench silo would make it harder for farmers to adopt. Under such circumstances, for long term use the bunkers would be a good choice for silage making with comparable silage quality as that of trench silo. Silage inoculants certainly improved the silage quality and it is highly recommended to use such additives for silage making and these additives are not that costly. Buffalo calves raised on fresh corn fodder and three different silages showed similar daily weight gain. Further growth trials on buffalo calves with varying levels of concentrate feeding along with silage are suggested to investigate silage feeding in calves. Corn silage fed lactating buffaloes had lower dry matter intake and total milk yield, but higher total solids as compared to fresh fodder feeding. The future studies of silage feeding compared to different inclusion levels of fresh and dry roughage sources would add further to explore the economic implications of silage feeding.
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Chemical, Microbiological And Toxicological Evaluation Of Textile Dyeing Industry Wastewater
Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: Exposure to complex mixtures like textile effluent poses risks to animal and human health such as mutations, genotoxicity, pathological lesions and oxidative damage. The aim of the present study was to quantify metals and identify organic pollutants in untreated textile dyeing industry wastewater, to determine the bacterial load of wastewater, isolate and identify heavy metals tolerant bacteria and to determine its mutagenic, genotoxic and cytotoxic potential, influence on normal physiology and effects on oxidative stress biomarkers in effluent exposed rats.
Metal analysis through AAS revealed presence of high amounts of zinc, copper, chromium, iron, arsenic and mercury in industrial effluent. Various organic pollutants such as chlorpyrifos, cucurbitacin-b and phthalates were identified by screening through GC-MS.
Microbiological evaluation of textile dyeing industry wastewater revealed a high bacterial load. Different bacteria isolated from wastewater such as Staphylococcus aureus, Pseudomonas aeruginosa, Corynebacterium xerosis, Bacillus megaterium, Staphyoloccus epidermidis and Micrococcus varians exhibited resistance to Cr and Cu salts and antibiotics to varying degree.
Ames test with/without enzyme activation and MTT assay showed strong association of industrial effluent with mutagenicity and cytotoxicity respectively. Bacterial reverse mutation assay revealed that the mutagenicity of textile dyeing industry wastewater decreased with increase in dilution of wastewater.
In-vitro comet assay revealed the evidence of high oxidative DNA damage induced by textile wastewater. Wastewater exhibited concentration dependent genotoxicity in sheep
peripheral lymphocytes. When Wistar rats were exposed to industrial effluent in different dilutions for 60 days, then activities of total superoxide dismutase and catalase and hydrogen peroxide concentration were found to be significantly lower in kidney, liver and blood/ plasma of effluent exposed rats than control. Vitamin C at a dose of 50mg/Kg/day significantly reduced oxidative effects of effluent in rats. Industrial effluents may decrease activities of T-SOD and CAT and concentration of H2O2 in liver, kidney and blood/plasma of Wistar rats. Vitamin C may have a possible ameliorating effect on industrial effluent induced oxidative stress in Wistar rats.
Wastewater exposed rats exhibited necrosis of epithelial cells of nephron, pulmonary emphysema, and inflammation of the lungs, degradation and infiltration of cardiac myocytes, fibrosis of the liver, damage to the intestinal mucosa and sloughing off epithelial cells from the intestinal lumen.
This study concludes that untreated textile dyeing wastewater being a complex mixture of inorganic and organic pollutants may be highly eco-toxic and may contaminate of the environment via continuous release of various organic and inorganic pollutants.
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Bats (Chiroptera: Mammalia) Of Malakand Division, Pakistan
Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: The present study was conducted from 2010 to 2013 in three districts (Malakand, Dir and Swat) of Malakand Division. A total of 49 stations were sampled for bats where total 1982 bats were recorded. A total of 21 species of bats belonging to six families, fourteen genera were recorded. These includes the Indian flying fox (Pteropus giganteus), the greater short-nosed fruit bat (Cynopterus sphinx), the fulvous fruit bat (Rousettus leschenaultia), the greater mouse-tailed bat (Rhinopoma microphyllum), the lesser mouse tailed bat (Rhinopoma hardwickii), the greater false vampire (Megaderma lyra), the greater horseshoe bat (Rhinolophus ferrumequinum), the Blyth‟s horseshoe bat (Rhinolophus Lepidus), the fulvous leaf-nosed bat (Hipposideros fulvus), the Hodgson‟s bat (Myotis formosus), the Asian barbastelle (Barbastella leucomelas), the Asiatic greater yellow house bat (Scotophilus heathii), the Asiatic lesser yellow house bat (Scotophilus kuhlii), the serotine (Eptesicus serotinus), the common pipistrelle (Pipistrellus pipistrellus), the javan pipistrelle (Pipistrellus javanicus), the coromandel pipistrelle (Pipistrellus coromandra), the least pipistrelle (Pipistrellus tenuis), the Dormer‟s bat (Pipistrellus dormeri), the desert yellow bat (Scotoecus pallidus) and the Schreiber‟s long-fingered bat (Miniopterus fuliginosus) were recorded throughout the study area.
M. formosus was common to all the three districts while B. leucomelas and P. pipistrellus were captured only from Dir district. The Hodgson‟s bat (M. formosus) and the Schreiber‟s long-fingered bat (M. fuliginosus) were captured from Malakand and Swat districts. The skeleton of C. sphinx was recorded only from adjacent area of Malakand district. The Indian flying fox (Pteropus giganteus) was not previously recorded from Khyber Pakhtunkhwa while it has been reported from Punjab and Sindh province of the country. There are only six species which has
previously been reported from Khyber Pakhtunkhwa while thirteen bats were newly recorded from the study area. Only two bats were newly recorded for the first time in the country.
The mean forearm length of the three P. giganteus was 152.23 mm ± 3.72 (SD). The mean greatest skull length was 65.96 mm ± 1.42 (SD). The maxillary toothrow length was 24.91 mm ± 0.84 (SD). The mandible and mandibular toothrow length were 50.78 mm ± 0.87 (SD) and 27.41 mm ± 0.66 (SD), respectively.
The thumb and forearm length of one C. sphinx was 25.80 mm and 65.48 mm, respectively. The greatest length of skull was 32.20 mm. The maxillary and mandibular toothrow length were 10.86 mm and 12.64 mm. The mandible was 24.75 mm long.
The mean forearm and thumb of R. leschenaultii was 80.23 mm ± 3.26 (SD) and 27.79 mm ± 1.22 (SD), long, respectively. The mean greatest skull length was 36.97 mm ± 1.11 (SD). The mean mandible, maxillary and mandibular toothrow length were 28.95 mm ± 0.90 (SD), 14.08 mm ± 0.44 (SD) and 15.51 mm ± 0.47 (SD), respectively.
Mean thumb and forearm length of three R. microphyllum was 8.80 mm ± 0.95 (SD) and 67.45 mm ± 4.60 (SD), respectively. The mean greatest length of skull was 20.15 mm ± 0.64 (SD). The mandible, maxillary and mandibular toothrow length were 7.30 mm ± 0.18 (SD), 8.11 mm ± 0.11 (SD) and 14.38 mm ± 0.63 (SD), respectively.
Mean thumb and forearm length of R. hardwickii was 8.23 mm ± 0.38 (SD) and 59.90 mm ± 1.21 (SD), respectively. The mean greatest length of skull of the four specimens was 18.20 mm ± 0.48 (SD). The maxillary and mandibular toothrow length were 6.08 mm ± 0.07 (SD) and 6.72 mm ± 0.13 (SD), respectively. The mandible length was measured as 12.38 mm ± 0.0.23 (SD).
Mean thumb and forearm length of M. lyra was 11.80 mm ± 0.44 (SD) and 70.06 mm ± 0.69 (SD), respectively. Mean greatest length of skull of the three specimens was 29.60 mm ± 0.46
(SD). The maxillary toothrow length was 11.40 mm ± 0.10 (SD). The mandibular toothrow length was 11.94 mm ± 0.04 (SD). The mandible length was measured as 20.04 mm ± 0.03 (SD).
Mean thumb and forearm length of R. ferrumequinum was 4.01 mm ± 0.01 (SD) and 60.01 mm ± 1.41 (SD), respectively. The mean greatest length of skull of the two specimens was 23.35 mm ± 0.20 (SD). The maxillary toothrow length was 9.18 mm ± 0.02 (SD). The mandibular toothrow length was 9.86 mm ± 0.01 (SD). The mandible length was measured as 16.33 mm ± 0.13 (SD).
The mean thumb and forearm length of R. lepidus was 3.87 mm ±0.13 (SD) and 38.02 mm ± 0.63 (SD), respectively. The mean greatest length of skull of the two specimens was 15.94 mm ± 0.15 (SD). The maxillary toothrow length was 5.86 mm ± 0.02 (SD). The mandibular toothrow length was 6.57 mm ± 0.64 (SD). The mandible length was measured as 10.34 mm ± 0.04 (SD).
Mean thumb and forearm length of H. fulvus was 4.91 mm ± 0.17 (SD) and 41.41 mm ± 0.97 (SD), respectively. The mean greatest length of skull of the thirteen specimens was 18.45 mm ± 0.16 (SD). The maxillary toothrow length was 6.50 mm ± 0.14 (SD). The mandibular toothrow length was 6.96 mm ± 0.18 (SD). The mandible length was measured as 11.73 mm ± 0.14 (SD).
Mean thumb and forearm length of M. formosus was 9.26 mm ± 0.70 (SD) and 48.74 mm ± 2.02 (SD), respectively. The mean greatest length of skull of the three specimens was 17.81 mm ± 0.12 (SD). The maxillary toothrow length was 7.15 mm ± 0.05 (SD). The mandibular toothrow length was 7.80 mm ± 0.05 (SD). The mandible length was measured as 13.85 mm ± 0.07 (SD).
Thumb and forearm length of B. leucomelas was 5.65 mm and 42.88 mm, respectively. The tragus height was 10.32 mm. The greatest length of skull of a single specimen was 15.87 mm. The maxillary toothrow length was 4.91 mm. The mandibular toothrow length was 5.43 mm. The mandible length was measured as 10.02 mm.
Mean thumb and forearm length of S. heathii was 9.06 mm ± 0.41 (SD) and 62.25 mm ± 1.76 (SD), respectively. The mean greatest length of skull of the nine specimens was 23.12 mm ± 0.46 (SD). The maxillary toothrow length was 7.87 mm ± 0.16 (SD). The mandibular toothrow length was 8.93 mm ± 0.16 (SD). The mandible length was measured as 16.62 mm ± 0.19 (SD).
Mean thumb and forearm length of S. kuhlii was 7.01 mm ± 1.41 (SD) and 50.06 mm ± 7.13 (SD), respectively. The mean greatest length of skull of the two specimens was 19.24 mm ± 0.71 (SD). The maxillary toothrow length was 6.49 mm ± 0.11 (SD). The mandibular toothrow length was 7.42 mm ± 0.01 (SD). The mandible length was measured as 13.78 mm ± 0.47 (SD).
Mean thumb and forearm length of E. serotinus was 8.92 mm ± 0.32 (SD) and 53.37 mm ± 1.39 (SD), respectively. The mean greatest length of skull of the fifteen specimens was 21.40 mm ± 0.70 (SD). The maxillary toothrow length was 7.84 mm ± 0.21 (SD). The mandibular toothrow length was 9.28 mm ± 1.95 (SD). The mandible length was measured as 15.51 mm ± 1.94 (SD).
Thumb and forearm length of P. pipistrellus was 4.01 mm and 31.06 mm, respectively. The greatest length of skull of a single specimen was 12.14 mm. The maxillary toothrow length was 4.22 mm. The mandibular toothrow length was 4.45 mm. The mandible length was measured as 8.27 mm.
Thumb and forearm length of P. javanicus was 4.02 mm and 32.01 mm, respectively. The greatest length of skull of a single specimen was 13.13 mm. The maxillary toothrow length was 4.60 mm. The mandibular toothrow length was 5.20 mm. The mandible length was measured as 9.46 mm.
Mean thumb and forearm length of P. coromandra was 4.70 mm ± 0.45 (SD) and 32.28 mm ± 1.17 (SD), respectively. The mean greatest length of skull of the eight specimens was 12.67 mm
± 0.40 (SD). The maxillary toothrow length was 4.44 mm ± 0.24 (SD). The mandibular toothrow length was 4.74 mm ± 0.23 (SD). The mandible length was measured as 9.13 mm ± 0.46 (SD).
Mean thumb and forearm length of P. tenuis was 4.43 mm ± 0.47 (SD) and 29.24 mm ± 1.03 (SD), respectively. The mean greatest length of skull of the 23 specimens was 11.56 mm ± 0.25 (SD). The maxillary toothrow length was 3.87 mm ± 0.09 (SD). The mandibular toothrow length was 4.10 mm ± 0.06 (SD). The mandible length was measured as 7.89 mm ± 0.60 (SD).
Mean thumb and forearm length of P. dormeri was 5.28 mm ± 0.70 (SD) and 34.30 mm ± 1.25 (SD), respectively. The mean greatest length of the skull was 13.77 mm ± 0.11 (SD). The mandible, maxillary and mandibular toothrow length were measured as 10.53 mm ± 0.09 (SD), 5.33 mm ± 0.02 (SD) and 5.56 mm ± 0.07 (SD), respectively.
Mean thumb and forearm length of S. pallidus was 6.26 mm ± 0.41 (SD) and 36.83 mm ± 0.42 (SD), respectively. The mean greatest length of skull of the twenty two specimens was 15.00 mm ± 0.26 (SD). The maxillary toothrow length was 5.66 mm ± 0.10 (SD). The mandible and mandibular toothrow length were 11.35 mm ± 0.23 (SD) and 6.11 mm ± 0.12 (SD), respectively.
Mean thumb and forearm length of M. fuliginosus bat was 6.61 mm ± 0.43 (SD) and 37.59 mm ± 5.37 (SD), respectively. The mean greatest length of skull of the six specimens was 14.48 mm ± 0.58 (SD). The maxillary toothrow length was 5.32 mm ± 0.39 (SD). The mandible and mandibular toothrow length were 10.54 mm ± 0.65 (SD) and 5.71 mm ± 0.49 (SD), respectively.
1. Bat surveys. This is the first extensive exploration of that small portion of the Khyber Pakhtunkhwa which comprises of only three districts of Malakand Division i.e. Malakand, Dir and Swat. Although more focus remained towards Malakand district, six families, fourteen genera, twenty one species were identified. Moreover, two new country
records (Myotis formosus and Miniopterus fuliginosis) were also explored. Further bat surveys in poorly surveyed parts of the country especially in KPK and Baluchistan may result in identification of some other new bat taxa. More bat surveys involving greater field efforts may also confirm the presence or absence of those already described from the country.
2. Distribution ranges and species specific habitat analysis. Presence of thirteen new locality records (Pteropus giganteus, Cynopterus sphinx, Rhinopoma hardwickii, Megaderma lyra, Rhinolophus Lepidus, Hipposideros fulvus, Barbastella leucomelas, Scotophilus heathii, Scotophilus kuhlii, Eptesicus serotinus, Pipistrellus javanicus, Pipistrellus dormeri and Scotoecus pallidus) and two new country records (Myotis formosus and Miniopterus fuliginosis) gives credence to the idea that distribution ranges of most of the bat species has change over the past sixty years. Thus serious scientific studies are needed to redefine distribution ranges and identify species specific habitats using global positioning system and radio-telemetric studies.
3. Reconfirmation of bat taxonomy. Genetic analysis of none of the bat species of the country has been made using molecular markers thus leaving behind a chance to doubt identification of cryptic bat species. Thus molecular genetic studies of all the bat species of the country is highly recommended which may also lead to the discovery of such bat taxa which are new to science.
4. Bat call library. The only bat detector (Patterson D 1000X) present in the country fell down from my hand in a water body and became out of order. So none of the bat could be recorded. Bat call analysis has boosted bat identification throughout the world but the
lack of such sophisticated equipment in the country has become a major bottle neck in the establishment of a bat call library.
5. Awareness campaigns. Majority of the countrymen are unaware of the ecological services rendered by bats. Khyber Pakhtunkhwa is the major fruit growing region of the country. Based on misperceptions, the locals consider all bats as vermin and kill them ruthlessly. Conservation education to highlight the significance of bats must be included in the curriculum of children at primary school level so that they may adopt a pro-conservation attitude in the first few years of their personality building.
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Chemical Microbiological And Toxicological Evaluation Of Pharmaceutical Effluent Wastewater
Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: Pharmaceutical effluent being a complex mixture of drugs and heavy metals may affect human health exhibiting a strong potential of mutagenicity, carcinogenicity, cytotoxicity and oxidative stress induction along with pathological changes in various organs of the body. The current study was focused to quantify the presence of heavy metals, detection of various drugs, determining the bacterial load along with isolation and identification of different bacteria and assessment of the mutagenic and genotoxic, cytotoxic and oxidative stress induction of pharmaceutical effluent wastewater when exposed to sheep lymphocytes, Salmonella typhimurium strains, cell lines and rats respectively.
Atomic absorption spectrophotometer was used to quantify heavy metals and showed the presence of arsenic, chromium, lead and iron in concentrations above the normal limits recommended by WHO and EPA. Gas Chromatograph mass spectrophotometer analysis shown the presence of digitoxin, lignocaine, caffeine and trimethoprim and various other organic pollutants.
Microbiological evaluation showed a high bacterial load in the pharmaceutical waste water. Several bacteria were also found in PEW in the presence of different drugs and heavy metals. Aeromonas sobria, Micrococcus varians, Staphyoloccus epidermidis, Staphylococcus aureus, Bacillus megaterium showed tolerance to potassium di chromate and copper sulphate and resistance to various antibiotic discs.
Ames assay revealed a strong mutagenic potential with and without the presence of metabolic activation mixtures. A concentration dependent effect was observed when samples were tested with increasing dilution factor.
MTT assay and comet assay also showed a concentration dependent effect. The BHK-21 cell line was used to evaluate cytotoxicity and cell viability decreased with increasing concentration of PEW. Sheep lymphocytes used in comet assay exhibited a concentration dependent DNA damage.
Different antioxidant enzymes were also evaluated. Rats were exposed to PEW at different concentrations and following 60 days oral exposure, rats were evaluated for the presence of total superoxide dismutase, catalase and hydrogen peroxide in kidney, liver and plasma. Exposure to Pharmaceutical waste water significantly decreased the (TSOD), (CAT) and (H2O2) levels in plasma, liver and kidney. Treatment with Vitamin E significantly ameliorated the levels of enzymes.
Exposed rats were also evaluated for any pathological changes. Coagulative necrosis of renal epithelial cells were observed along with severe degeneration and cellular swelling in hepatocytes of hepatic cord.
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Effects And Remedial Measures Of Aflatoxin B1 On Bovine Calves In Punjab
Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: Aflatoxins B1 are most toxic metabolites produced by Aspergillus fungi in/on foods and feeds, probably best known and most intensively researched aflatoxins globally. AFB1 have been associated with several diseases, e.g. aflatoxicosis in livestock, pets including humans throughout the world. Occurrence of AFB1 is influenced by certain environmental factors like geographic location, agro-economic practices and susceptibility of feed commodities to fungal invasion during pre-harvest, storage, and processing periods. AFB1 has grabbed greater attention than any other mycotoxins due to their demonstrated potent carcinogenic effect in susceptible animals and their acute toxicological effects in humans. As the absolute safety will be never achieved, most of the world struggled to limit aflatoxin exposure by imposing regulations on feed commodities.
So, in this study, we had collected 67 concentrated samples, thirty six samples from Gujranwala and thirty one from Kasur to examine the occurrence of aflatoxin B1. The aims of this study were to investigate the aflatoxin B1 in calf feed, effect of different concentrations of aflatoxin B1 on productive performance of calves and determine the comparative efficacy of commercially available mycotoxin binders and liver tonics against AFB1 in bovine calves. Feed samples were obtained from different livestock farms and cattle feed mills, toxin levels in each feed sample were determined by HPLC. AFB1 level was higher at feed mills (40.33±2.21 ppb and 49.0±1.95 ppb) than farms (34.96±2.65 ppb and 44.95±2.41 ppb) both in Gujranwala and Kasur respectively. Fungus was isolated and grown on Sabouraud’s dextrose agar on the basis of microscopic characters and species within genus characterized by colony characters/macroscopic characters, mostly Aspergillus species was present in the feed samples which produce mycotoxins. The second most prevalent species were the Fusarium. Mucor and the Pencillium were respectively third and fourth in number. Our results have shown that Alternaria was not present in Gujranwala and Rhizopus was absent in the feed samples collected from the Kasur. Out of mycotoxin contaminated concentrate feed samples, the highest frequency of Aspergillus (43.3%) was observed, followed by Fusaram (38.8%), Mucor (8.9%), Penicillium(5.9%), Rhizopus (1.5%) and Alternaria species (1.5%).
Our results also indicated that growth of Aspergillus spp. can be minimized by controlling the different factors like pH, temperature, light and humidity, which are essential for the proper growth and development. The antifungal activity of methanolic extract of clove, neem and garlic was also determined in which maximum MIC showed by garlic.
Thirty six bovine calves of 6 to 12 months of age were kept in UVAS, Pattoki campus (Ravi Campus) .in four different replicates having 9 animals each. Different concentrations, i.e. 0.6 mg/kg, 0.8 mg/kg and 1.0 mg/kg was administered along with concentrated feed and check out productive performance along with physiological profile. The most pathological concentration of aflatoxin B1 in experiment number 3 was given to the two groups of bovine calves along with two different commercially available mycotoxin binders i.e. Yeast based and second one was clay based HSCAS mycotoxin binder at recommended doses. Efficacy of mycotoxin binders on feed samples was analyzed by using HPLC and also evaluates the productive performance of the animals.Efficacy of two liver tonics i.e.silymarin and choline chloride was observed on CBC, LFT and RFT of bovine calves.
Present study has clearly displayed the adverse effect of aflatoxin B1 on feed consumption, hematological and serum biochemical parameters related to liver and kidney in bovine calf. Results indicated that HSCAS mycotoxin adsorbent was able to fully detoxify aflatoxin B1. Silymarin had great impact on the liver to cope the adverse effects of the AFB1 as compared to the choline chloride, which was proved with the help of CBC, LFT and RFT.
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Identification And Expression Analysis Of Genes Involved In Obsessive Compulsive Disorder In Pakistani Population
Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: The background of this study is that WHO reports that psychiatry disorders affect worldwide 0.8 to 2% population. Anxiety illnesses are a class of illness associated with unreasonable and disturbing sensation of fear and tension. There are several types of anxiety disorders, such as panic disorder, agoraphobia, specific phobia, social phobia, OCD. Obsessive-compulsive disorder is a chronic disabling condition. OCD is characterized by repetitive, intrusive thoughts, images, and impulses and by repetitive, ritualistic physical or mental acts performed to reduce the attendant anxiety. The severity of OCD depends on the amount obsessions and compulsions. The Yale-Brown Obsessive Compulsive Scale (Y-BOCS) is a reliable and consistent scoring system that can be used to categorize OCD. The major genes involve in OCD are SLC6A4, BDNF, SLC1A1 and COMT genes.
The study was enrolled patients treated for OCD. Blood samples have been collected from the patients. DNA extracted from fresh blood. Primers were designed. Then DNA amplification have done by Bio-Rad thermal cycler. Then gel electrophoresis was done for PCR product quantification. PCR products precipitated and sequenced. SNPs were identified. Real-time quantitative RT-PCR was performed for each sample with TaqMan Universal PCR mastermix which showed down regulation of COMT gene in OCD patients in Pakistani population.
The aim of this study was SNP identification in Pakistani Population in Obsessive Compulsive disorder and to analyze the gene expression of COMT gene involved in OCD in Pakistani Population.
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Molecular And Serological Characterization Of Soilborne Francisella Tularensis
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Publisher: 2016 Dissertation note: Tularemia, caused by Francisella tularensis (F. tularensis ), is a zoonotic disease transmitted through contact with infected animals and contaminated environment. The disease has been reported from many countries of the world but no study has been done in Pakistan. In the current project, a total of 2280 soil samples representing 456 villages of eight districts of Punjab province were collected from way-points having human-animal interaction, processed for genomic extraction and tested through real time PCR for presence or absence of F. tularensis. Association of risk factors was determined from data such as gender and age of animals, plough method, irrigation system, fertilizer type used, availability of veterinary services, level of farmer education, physical and chemical composition of the soil. Moreover, sero-prevalence against F. tularensis in cattle, buffaloes, sheep and goats was determined using ELISA. Seventy four soil samples (3.24 percent) were found positive for F. tularensis. Phylogenetic analysis showed 100 percent similarity index with F. tularensis sub specie holarctica reported from other regions like USA, Sweden, Spain, Turkey and Germany. Presence of F. tularensis in soil showed negative association with increase in number of human density (0.7159; 0.3834-0.2054). Prevalence of anti- F. tularensis ELISA antibodies were significantly higher (p<0.05) in large ruminants (cattle and buffalo) as compared to small ruminants (goat and sheep). Age and gender-wise analyses showed non-significant differences (p>0.05) between small and large ruminants. Whereas, rain-irrigation system (2.96: 1.35- 6.48), lack of veterinary services (4.77:1.26-18.03) and use of organic fertilizer (5.3: 11.38- 20.39) have positive association with prevalence of anti- F. tularensis ELISA antibodies in the serum. Sero-prevalence of Ft in the animals has significant association with quantity of clay in soil (p<0.05). A conventional PCR based test has also been optimized for
detection of F. tularensis using tul4 gene specific primers. Specificity of primer showed F. tularensis detection in soil DNA in the presence of other cross-reactive organism. Sensitivity was determined in two fold dilutions with detection limit of up to 320 pg/μL. Utilizing pET28a vector, a construct was prepared containing transformed tul4 gene (450bp) showing 100 percent sequence homology to query gene sequence. For manufacturing diagnostic assays especially in developing countries where availability of BSL-3 facilities and positive control reagents is an issue, provision of tul4 gene based constructs in vector can act as positive control and biosafe to use.
It is recommended that similar studies may be done in other parts of Pakistan to have spatial distribution of F. tularensis all over Pakistan. In future studies, other sources of transmission like water, ticks and rodents may be considered with soil for complete analysis. Transportation of whole genome of F. tularensis has been prohibited by Russian government, ATCC and CDC, WHO is working on designing a complete protocol for transportation of this bacteria or genome to other countries. Under such situation, conventional PCR optimization can be done for diagnosis of F. tularensis and pET28+tul4 constructs, developed in this study, can be used as a PCR positive control reagents.
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Effect Of Colchicine On Cellular And Humoral Immune Responses In Mice
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Publisher: 2016 Dissertation note: Colchicine is a medication that treats gout. It is a natural product and secondary metabolite, originally extracted from plants Colchicum autumnale .It causes modulation of chemokine and prostanoid production and inhibition of neutrophil and endothelial cell adhesion molecules by which it interferes with the initiation and amplification of the joint inflammation.
The present study is designed to evaluate the effects of colchicine on cellular and humoral immunity in mice. There were five groups for each assay i.e. group I (negative control), positive control and three colchicine treated group II (40μg/kg), group III (80μg/kg) and group IV (160μg/kg). The number of mice in each group was five to eight. All these groups were administered doses intraperitoneally. To determine the effect of colchicine on cell mediated immunity , delayed type hypersensitivity (DTH) assay, macrophage engulfment assay, cyclophosphamide induced neutropenic test and nitric oxide production was performed .DTH was performed by measuring skin thickness. DTH showed significant difference (P<0.001) of negative control to colchicine treated groups 40μg/kg, 80μg/kg and 160μg/kg. With increasing dose, there was decrease in skin thickness of the mice. Highest reduction of skin was found at 160μg/kg. Macrophage engulfment assay was performed to evaluate the effect of macrophage induced phagocytosis. There was significant ( P <0.001) difference of engulfment of SRBCs by macrophages with negative control to colchicine treated group II (40μg/kg), group III(80μg/kg) and group IV(160μg/kg) groups. There was significant difference of engulfment of macrophages at 45 and 90 minutes.
Cyclophosphamide induced neutropenic test was performed to assess the effect of colchicine on total leukocyte count (TLC) and differential leukocyte count (DLC). There was
reduction of TLC to about 45.3% in control to 48.3%, 54.68% and 65.42% in group II (40μg/kg), group III (80 μg/ kg) and group IV (160μg/kg) respectively when these were compared with primary values of TLC. There was significant difference of reduction in the neutrophil count of negative control 1057 (±120) to 902 (±67) in group II (40μg/kg), 734(±69) in group III (80 μg/ kg) and 609 (±71) in group IV (160μg/kg) of doses of colchicine. This test showed that with the increasing dose of colchicine, there was significant (P<0.001) difference of TLC count and neutrophil count.
Nitric oxide (NO) production by macrophages was performed for measuring different concentrations of nitric oxide produced. There was significant difference (P<0.001) in NO production by macrophages alone and LPS stimulated between negative control to group II (40 μg /kg), group III (80μg/kg), group IV (160μg/kg) of colchicine. With increasing dose, there was significant reduction in production of NO. There was significant P<0.0001 reduction in body weight andspleen weight difference of mice in different groups of colchicine treated 40μg/kg, 80μg/kg and 160μg/kg from negative control after treatment. There was difference of weight of Thymus of group II (40 μg/kg), group III (80μg/kg) and group IV (160μg/kg) but difference was statistically not significant. There were no histopathological changes observed in spleen and Thymus at 40μg/kg and 80μg/kg doses of colchicine. At 160μg/kg dose, increase in thickness of trabecular was seen .due to edema in the spleen. For evaluation of colchicine effect on humoral immunity, haemagglutination assay, mice lethality test and Jerne hemolytic plaque formation were performed. Haemagglutination assay (HA) was performed by using red blood cells injected intraperitoneally in mice to measure antibody titer. There was significant difference of (P >0.001) to colchicine treated group II (40μg/kg), group III (80μg/kg) and group IV (160μg/kg)with group I (negative control).With the increasing dose, there was reduction in the
HA titer. Mice lethality test was performed by testing immune response of the mice to the challenge infection of P.multocida. It was performed by comparing mortality ratio of mice after administration of drug. There was no death of mice in the negative control group in which there was administration of PBS and vaccine. At 40μg/kg dose of colchicine, there was 50% mortality ratio. At 80μg/kg dose of colchicine 75% mortality ratio was observed. Maximum mortality ratio was observed at the 160μg/kg colchicine dose i.e. 100%.
Jerne plaque formation test was performed and plaques formed was enumerated and recorded as the number of plaque forming cells (PFCs) per million cells. There was significant difference (P<0.001) of reduction in number of plaques from negative control to all doses of colchicine 40 μg/kg, 80 μg/kg and 160μg/kg. Antibody formation was decreased with increasing the dose of colchicine. Therefore, it is concluded that colchicine suppresses the cellular and humoral responses in mice.
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Epidemiology, Zoonotic Potential, Molecular Characterization And Therapeutic Trial Of Leptospirosis In Horses
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Publisher: 2016 Dissertation note: Leptospirosis is an important zoonotic disease. It affects a wide range of mammals, fish and even a few reptiles. It is caused by Leptospira interrogans, having more than 250 serovars, distributed geographically throughout the world. In horses, Leptospira interrogans causes liver and renal abnormalities, ERU, and reproductive disorders in mares like abortion, perinatal death and still birth. It is transmitted to human beings, working with live or dead tissue of infected horses and through surfaces contaminated with urine of carrier or infected animals. In humans, it causes influenza like illness and death in severe cases. Serological testing, bacterial culture and molecular techniques are used for the diagnosis of disease. This study was aimed at estimating the seroprevalence of Leptospira spp. in horses and humans of three climatically distinct regions of Punjab, Pakistan. Furthermore, molecular biology techniques were employed for the confirmed diagnosis of equine leptospirosis and therapeutic efficacy of ampicillin and adhatoda vasica was analyzed against disease. It was the very first study in Pakistan conducted to explore equine leptospirosis in the country.
During this study, 384 horse blood samples and epidemiological data were collected from three climatically distinct regions, viz;Rawalpindi, Lahore and Bahawalpur (128 from each study area) and were subjected to ELISA to determine seroprevalence of Leptospira. Results showed overall prevalence of 33.85% in Punjab with highest prevalence in Rawalpindi (40.62%) which experienced highest rainfall, followed by Lahore (38.28%), and least in Bahawalpur (22.65%). Risk factor analysis showed that age, gender, living area, herd size, water source, exposure to rodents and floods, feeding practices and usage of animals were found significantly associated with the disease. To study the seroprevalence of human leptospira, 360 human blood samples were collected (120 from each study area). Epidemiological data on pre-structured questionnaire
were collected from all the participants of study. All the samples were subjected to ELISA and results showed overall prevalence of 40.83%, with highest seroprevalence in Rawalpindi (50.83%), followed by Lahore (38.28%) and least in Bahawalpur (27.50%). Age, gender, occupational and living area, water recreational activities, occupation, exposure to floods, educational status and history of wound were significantly associated risk factors while use of PPE during work was deterrent.
During this study, 65 ELISA positive horse samples were subjected to molecular biology diagnostic technique PCR for the molecular characterization of equine leptospirosis in country. After DNA extraction, PCR was performed using primer sets specific for 16S rRNA gene, which yielded a fragment of length 306bp after gel electrophoresis. Out of 65 tested samples, 20 samples (30.76%) were PCR positive and was further sequenced and phylogenetic tree was constructed. Dendogram showed the sequenced samples were related to pathogenic Leptospira interrogans, revealing potential of 16S rRNA primer sets for the detection of eqine leptospirosis in country. Dendogram further showed closed resemblance of analyzed samples with serovar Icterohemmorhagae, Australis and Autumunalis which are dominant serovars in India, Iran and China, the neighboring countries of Pakistan. Therapeutic efficacy of ampicillin and AV was studied by analyzing the hematology, liver function test, renal function tests and serum mineral levels at day 0 (pre-treatment), 7, 21 and 35 (post-treatment). Results showed that all the tested parameters were changed significantly during infection and significant improvement was observed after treatment. Ampicillin was instrumental in revealing hematological abnormalities while AV played important role in normalizing the liver and renal insufficiency. After treatment ampicillin treated 58.33% of animals and AV treated 41.66% of animals.
This first ever study of equine leptospirosis in country uncovers the high prevalence rates in horses and humans and raises a need for control strategies to prevent the transmission and spread of the disease. It also highlights the potential of molecular biology techniques for the confirmed diagnosis of equine leptospirosis and explores options for designing better specie specific treatment regimes for the disease.
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Pathogenesis Of Aflatoxin B1 In Quails Under Experimental Conditions And Detoxification By Biological And Chemical Means
Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: Secondary metabolites of certain fungi produce toxins under favorable conditions especially while growing on different food grains. Mycotoxins are among major threats to growing poultry industry and human beings. Aflatoxins are closely related, biologically active fungal metabolites and commonly produced by Aspergillus species.
A research was carried out to evaluate the ability of Aspergillus flavus for Aflatoxin B1 production using rice, wheat and maize as substrates. Lethal effects on growth performance parameters, hematological and histopathological of graded doses of aflatoxin B1 in quails under experimental conditions were observed. Effect of Aflatoxin B1 on humoral immune response to Newcastle Disease virus vaccine in quails were determined. Biological detoxification of Aflatoxin B1 by Saccharomyces servisiae was evaluated in quails. Comparative evaluations of different commercially available toxin binders were checked. All these experiments were carried out till the six weeks (42 days).
Aspergillus flavus was identified on the basis of macroscopic and microscopic characteristics. Rice, wheat and maize grains was used as substrate to check the level of Aflatoxin B1 produced by inoculating an aqueous suspension of 106 spores/ml. Aflatoxin B1 checked by Thin Layer Chromatography (TLC) and quantified by High Performance Liquid Chromatography (HPLC).
Quails were reared under standard management conditions in five groups (A, B, C, D and E) having sixty each. Each group was further divided in two independent units. Diets offered to groups were control (without toxins), 0.25, 0.50, 1 and 2 mg Aflatoxin B1/kg feed. One unit of
each group was vaccinated with Newcastle Disease Virus (NDV) vaccine while other was not and studied the lethal effects on growth performance, blood parameters, immune response and histopathology of vital organs. At the end of the experiment, it was found that the deleterious effects of Aflatoxin B1 were dose and duration dependent. As the level of the toxin was increased, the lethal effects were prominent. The growth performance parameters including gain in body weight, feed intake and feed conversion ratio was adversely affected at high doses. The body weight gain was significantly reduced in Aflatoxin B1 treated groups as compared to control group. Similarly feed intake and feed conversion ratio were significantly different from the control group. The hematological studies exhibited that aflatoxin B1 significantly reduced the hemoglobin, packed cell volume and total leukocyte count whereas the erythrocyte sedimentation rate was significantly increased as compared to control group. The immune response against NDV vaccine was adversely effected in Aflatoxin B1 treated groups and values of Antibody titer in AFB1 were significantly low as compared to group A( control) In the second experiment, Saccharomyces cervisae (SC) dried powder was mixed in basal quail diet having 0.5mg Aflatoxin B1 for all experimental groups and control was without toxins. SC was added at levels of 0.5 gm, 1.0 gm and 2.0 gm /kg of feed. It was recorded that Saccharomyces cervisae (yeast) have the potential to remove the deleterious effects of Aflatoxin B1. Yeast effectively detoxified the Aflatoxin B1. The results recorded of growth performance and other parameters were non-significantly different from the control group. Chemical detoxification of Aflatoxin B1 was evaluated in quails using commercially available toxin binders. Toxin binders used were activated charcoal, kaoline, Myco AD and selenium plus vitamin E and mixed in basal quail diet having 0.5mg Aflatoxin B1 for all experimental groups and control was without toxins. The Myco AD and selenium plus vitamin E showed the highest detoxification potential as compared
to other chemical toxin binders. Groups E and F showed the results of growth performance, hematological, immune response and histopathological were non-significantly different from the control group (A). Kaolin was moderately detoxifying the toxin.
Presence of aflatoxin B1 in soft tissues was checked by TLC and quantified using HPLC. The liver exhibited the residues of Aflatoxin B1 at high doses of toxin. Group D and E rearing on feeds having 1mg AFB1 /Kg feed and 2mg AFB1 /Kg feed of toxin showed the residues of AFB1 in liver and kidney.
Statistical means for growth performance parameters, hematological, immune response and histopathological scores in each subunit of quails were analyzed by applying one way ANOVA and Duncans‟s Multiple Range (DMR) test at 95% probability. Aflatoxin B1 is lethal and lowers the performance of birds. The lethal effects can be detoxified by biological and chemical means to lower the economic losses to poultry industry. It can be concluded that biological detoxification is preferably better as compared to chemical detoxification.
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Expression, Purification Of Toxoplasma Rop18 Recombinant Protein And Its Antigenic And Immunogenic Trials In Mice
Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: Toxoplasma gondii is an obligate intracellular, apicomplexan parasite that infects all warm-blooded vertebrates, including mammals and birds. Human beings can be infected by ingestion of oocysts from cat feces or through the consumption of meat containing Toxoplasma gondii cysts. There are potential vaccines candidates among which ROP18 has its major role in host gene expression along with the modulatory effect on key regulators of the host immune system. Therefore in this study, ROP18 sequence was amplified from local T. gondii strain, recombinant ROP18 was expressed through recombinant DNA technology and this recombinant protein was then tested for its antigenicity and immunogenicity in a mouse model. Approximately 200 fecal samples were collected from domestic, wild and stray cats in and around city of Lahore, Pakistan. Oocysts of T. gondii from cat feces were identified by using light microscopy and flotation technique. The oocysts were measured by micrometry having diameter of 8-10 μm. Out of 200 fecal samples, only three were suspected for T. gondii through direct microscopic examination and flotation technique. From 3 fecal samples, genomic DNA was extracted using a stool DNA extraction kit. After DNA extraction, the 3 samples were confirmed and characterized by PCR and nested PCR by using B1 gene and SAG2 primer sets. Reference DNAs (RH) of toxoplasma were kindly provided by Dr. Henrik Vedel Nielsen (Statens Serum Institut, Denmark) and Dr. Jorge Enrique Gomez Marin (COLOMBIA, South America). For detection of the B1 gene of T. gondii, the diagnostic method was optimized to amplify a 529 base pair (bp) repetitive sequence by PCR using DNA extracted from cat feces. Then a nested PCR was employed using internal primers to amplify a 102 bp from 391 bp product. The SAG2 gene was targeted at 5 different regions to amplify 5 amplicons. Genotype analysis was done using SAG2 sequence by Dr.
Jorge Enrique Gomez Marin using 10 different markers. For amplification of ROP18, 54 sequences of the ROP18 gene retrieved from Genbank (National Center for Biotechnology Information (NCBI)) We used Geneious R8.1.6 software for sequence alignment and creating consensus sequence from all 54 ROP18 sequences. Primers were designed manually from the consensus sequence of ROP18. Primer pair namely ROP18-F 5‟ATCTAGAATGTTTTCGGTACAGCGG3‟ and ROP18-R Reverse 5‟TTCGAATTCTGTGTGGAGATGTTCC3‟ were designed to have restriction sites XbaI and HindIII respectively. The rop18 sequence was first cloned in pGMT easy vector system and then subcloned in pET28. BL21 competent cells were transformed with pET28-ROP18 and rROP18 was expression using IPTG for induction. The rROP18 was quantified through protein quantification kit (BCA). The rROP18 was formulated into nanospheres using PLGA as coating material. The Swiss-Webster mice were inoculated either intranasal or subcutaneous with rROP18 with or without montanide as adjuvant 3 times with 2 weeks interval. The blood was collected 2 weeks after each immunization. The control groups were inoculated with PLGA I/n or montanide S/c. For western blotting, ROP18 protein was electrophoresed on SDS-PAGE and blots were immune-blotted with the sera of immunized or infected mice. Bound antibodies were detected through Goat anti-mouse IgG–alkaline phosphatase conjugated. For evaluation of humoral response, ELISA plate was coated overnight at 4°C with rROP18 protein at 5μg/ml in 50mM sodium carbonate buffer (pH 9.6) @ 100 μl/ well. The absorbance of each sample was measured at OD 405 nm using ELISA (Bio-Tek, E-800, USA). Comparisons of quantitative values in the different groups were performed using ANOVA test, after checking the homogeneity of variances. Comparisons between groups for the antibody titre were performed by Dunn multiple range tests test. Comparisons were considered significant when a probability of equality was less than 5% (P<0.05). It was observed that rROP18 in nanospheres administered intranasal elicited
elevated responses of specific intestinal IgA and IgG2a as compared to other groups inoculated intranasally rROP18 alone or injected subcutaneously rROP18 adjuvanted in montanide. It was concluded that nanospheres of ROP18 would be a non-invasive approach to develop vaccination against toxoplasmosis. Further experiments are needed to conclude the cellular response of these nanospheres in a chronic mouse model.
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Molecular Identification And Treatment Of Theileriosis In Small Ruminants Of Northern Balochistan
Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: The present study was conducted to investigate the prevalence of Ovine and Caprine Theileriosis in Northern Highlands and Suleiman Mountain Region of Balochistan, Six thickly populated /union councils were included in the study area. Samples were collected from 2870 animals Sheep (n= 2200) and Goats (n= 670) for screening of the disease. The samples were collected and processed in Regional Disease Investigation Laboratories, Department of Livestock and Dairy Development Balochistan, T.B. Sanatorium Hospital Quetta and Center for Vaccinology, Bacteriology, The University of Balochistan, Quetta and Medicine Laboratory, Department of Clinical Medicine and Surgery, The University of Veterinary and Animal Sciences, Lahore. Data revealed 20.82% disease in sheep and 9.70%. in goats. The regional prevalence of theileriosis revealed 19.19% in Northern Highlands and 17.48% in Suleiman Mountain Region Chi-square analysis showed significant difference in the prevalence of disease in sheep and goats. The regional difference was not significantly different between two regions of Northern Balochistan. The comparison among union councils showed significant difference being highest prevalence (22.71%) in union council Kuchlak district Quetta followed by Aghberg (18.42%) and Hanna Urak (15.53%) in Northern highlands and Union Council Zangiwal Jogezai (19.83%) followed by Kach Amaqzai (16.30%) and Sinjavi (15.92%) in SMR. The disease prevalence when compared among 4 different breeds of sheep showed significant difference being highest in Karakul breed (34.62%) followed by Shinwari (24.54%), Bibrik (19.36%) and Harnai (16.40%). The highest prevalence of theileriosis in sheep and goats were observed in Summer season (30.30%) followed by Autumn 19.07%, Spring 14.52% and Winter
7.61%. Chi-square analysis of the data showed significant difference in the prevalence of the disease in different seasons of the year. The disease was also compared in three age groups of sheep and goats. The data showed 22.17% disease in adult animal group above 2 years of age followed by 15.85% in animals between 1-2 year and 7.99% in age group below one year. Statistically significant difference in all age groups was found in chi-square analysis. The sex wise prevalence of theileriosis revealed non-significant difference between male and female sheep and goats. Two different species of Theileria were reported by many researchers causing disease in sheep and goats. The PCR was carried out for the identification of Theileria species affecting sheep and goats in Balochistan. Two species specific sets of primers were designed using 18SRNA gene sequence to identify these two species of Theileria and the distribution among the two species of animals. The genomic DNA of two species of parasite was successfully amplified in positive samples. The assay was proved successful and we recommend for the prevalence surveys for theileriosis in sheep and goats. The data showed that the prevalence of T. lestoquardi was 73.80% in sheep and 69.23% was in goats in the target regions. It was found the T. lestoquardi was highly prevalent and causing theileriosis in small ruminants. The prevalence of T. ovis was 26.19% in sheep and 30.76% in goats respectively in the investigated animals; it was less than T. lestoquardi. It was concluded that both Theileria species were identified and found circulating in small ruminants in the target region of Balochistan. In the study we determined that PCR method based on 18S RNA gene could detect and differentiate T. ovis and T. lestoquardi.
Effect of theileriosis in sheep and goats on hemeto-biochemical parameters were studied included RBCs, Hb%, PCV, Platelets, WBCs, MCV, MCHC, AST, ALT, BUN, Bilirubin and Creatinine. Blood samples were collected from Theileria confirmed, diseased animals (sheep and
goats) along with equal number of healthy animals for comparison. In sheep RBCs, Hb%, PCV, WBCs, MCHC, AST, ALT and Creatinine values showed significant difference when compared with values of healthy animals. Significant (p<0.05) reduction was noted in measurement of RBCs, Hb%, PCV and MCHC whereas, AST, ALT and Creatinine showed significant increase in diseased animals. In goats affected with theileriosis showed significant decrease in RBCs count and Hb%. The values for AST, ALT and Creatinine were found significantly increased in diseased animals when compared with healthy control group of equal number of animals. In present study it was noted that Butalex intra muscularly at the rate of 2.5 mg/kg body weight is quite effective in eliminating the Theileria parasite from the blood of sheep and goats and treatment at the day 10 post treatment. Imizol was also found an effective treatment of theileriosis but less effective than Butalex.
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Epidemiology Of Influenza Virus H5n1 In Islamabad Capital Territory
Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: The poultry sector in Pakistan is the second largest industry that contributes to the national gross domestic products (GDP) and remains a major source of nutrition (protein and energy) for human population in Pakistan. Highly Pathogenic Avian Influenza (HPAI) outbreaks due to H5N1 virus in poultry have been recorded in over 62 countries, indicating the contagious nature of the disease and its potential to infect various avian species. These HPAI outbreaks in poultry have lead to killing/culling of around 120 million birds in various countries. During 2009, the Avian Influenza continues to occur in poultry in China, Hong Kong, India, Egypt, Nepal, Bangladesh and Canada . In Pakistan, an HPAI outbreak due to H7N3 virus was first observed in 1994-95 and those due to H9N2 virus in broiler and layer chickens were recorded between late 1990’s and early 2000. During the period between 2006 and 2008, poultry heavily suffered due to multiple outbreaks caused by H5N1 virus.
The country experienced several and severe HPAI subtype H5N1 outbreaks during 2006-2008 in commercial poultry farms mostly, causing mass economic losses. In Pakistan all the four poultry production system exists being identified by FAO. The present study was conducted in peri-urban areas of ICT Islamabad, capital of Pakistan. The objectives of the present study were to investigate the outbreaks due to HPAIV H5N1 in 2006-2007 in ICT and identify the pattern and trends of these outbreaks. For this purpose descriptive epidemiological study was conducted and data was collected on a predesigned questionnaire regarding farm demography, culling, morbidity and mortality. The result statistical analysis showed a significantly (P< 0.05) higher morbidity, mortality, case fatality and culling rate in layers farms than breeders and broilers respectively. Layers and breeders of old ages were mostly affected with having higher mortality and culling in comparison to younger age layer and breeder commercial farms. The mean morbidity and mortality rates ranged 57–95% and 5-43% correspondingly.
After the HPAIV H5N1 first reported outbreak in Pakistan in 2006 culling strategy was adopted after devastating outbreaks regularly reported from throughout the country. The reasons behind these emerging epidemics were unknown and several hypotheses were given birth after these outbreaks. Knowledge regarding potential risk factors responsible for HPAIV H5N1 epidemics in commercial poultry farms in Pakistan was lacking. Therefore we conducted a longitudinal cross sectional survey (1:1 matched case control study) to identify potential risk factors at farm level responsible for 2006-2007 HPAIV H5N1 infection in poultry in ICT. Information on farm characteristics, biosecurity practices and farm management were collected. Logistic regression model on data was used to unveil the potentially associated risk factors with cases (farms confirmed HPAI H5N1 Positive). Several candidate variables were studied and investigated for association. The results multivariable logistic regression showed that farm location such as in urban area (P<0.05: OR=18.50), wild birds entry (P<0.05: OR= 12.66) and farms situated in highly dense poultry populated area (P<0.05:OR=4.50) were found significantly associated with outbreaks of HPAIV H5N1 infection in commercial poultry farms during 2006-2007 epidemics in the study area.
Live bird markets (LBMs) are essential for poultry marketing in developing countries like Pakistan. One year active disease surveillance for influenza viruses in avian species in LBMs in ICT area was conducted in 2011. LBMs in Pakistan are typically urban that brings together many avian species produced by different suppliers. Which make LBMs in Pakistan a potential source of HPAIV viruses as well as other emerging poultry pathogens i.e. new castle disease virus,infectious bronchitis etc. The results of the present surveillance data showed that seroconversion against H5N1 and H9N2 is present in LBMs bird species which were isolated from different samples like serum, cloacal, nasal samples and organ samples.This indicates the continuous threat of AIV viruses circulating in the live bird markets set up of Pakistan.
Findings of these studies will help to tailor control and prevention measure against devastating outbreaks in future regarding the local circumstances of commercial poultry farms as well as in LBMs. These studies also succeeded to unveil the true reasons behind these devastating outbreaks and their higher impact on poultry industry. Such type of surveillance programs will be useful in future to investigate several emerging diseases and outbreaks in Pakistan and other developing countries.
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Molecular Epidemiology Of Mycobacterium At The Animal Human Interface And Its Co-Morbidity With Diabetes Mellitus
Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: Tuberculosis (TB) is a common and fatal infectious disease which has afflicted mankind for several millennia. At the moment, TB is positioned at number five when it comes to the most common causes of fatality worldwide. TB is curable if it is properly diagnosed and treated. In 2015, it was estimated that 1.5 million deaths (an equivalent of 4,000 deaths per day) and 9 million new TB cases have been reported. Diabetes Mellitus is also widely distributed and estimated to affect 366 million people by 2030. The co-morbidity of DM and TB is re-emerging because of the progressive epidemiology of both diseases especially in the developing countries. Endemicity of TB and DM is growing in developing countries because of low socio-economic status and poor living conditions.
In this study, a total of 500 tuberculosis positive patients were selected under TB DOTS program from five tertiary care hospitals of Lahore. Sputum samples were collected from all the enrolled patients and smear microscopy was performed for TB confirmation. Blood samples were collected from the same patients for screening of diabetes mellitus. Sputum samples were also processed for culture and drug sensitivity on LJ medium. Molecular identification by PCR technique was carried out on all positive cultured strains and results were compared with reference strain H37RV. For DNA sequencing, PCR products were sent to Singapore where sequencing was performed by Sanger method.
Data was compiled and variables including gender, age, drug resistance and treatment history and correlation among different variables was analyzed using chi-square test and Fischer’s exact test method at P-value of ≤0.05. SPSS (Statistical Package for Social Sciences, Version 20.0) was used for statistical analysis. The count data was statistically analyzed using
descriptive statistical tools. On screening for fasting blood sugar level, 74 (14.8%) patients were recorded as diabetics as well i.e. blood sugar level ≥ 126 mg/dl. Out of these 74 patients, 22 patients had previous history of diabetes whereas remaining 52 patients were newly diagnosed at the time of screening. The maximum distribution of TB-DM patients was found in age group > 57 years. Mean age of the group without DM was 39 years and with DM was 48 years. Coexistence of DM in TB patients was higher in males (62.2%) as compared to female study subjects. However, the gender difference is statistically non-significant (p value 0.243).
The distribution of education level revealed that out of the total participants, maximum number of patients (n=220) were illiterate and similar trend was observed in diabetic patients with 54 (73%) individuals belonging to the illiterate group of the subjects. There is statistically significant difference between existence of DM and literacy level in tuberculosis patients. Among social and behavioral risk factors in tuberculosis patients, majority of the patients were unemployed (24%) in TB-DM group. Significant correlation p value ≤ 0.05 was found between coexistence of TB-DM and tobacco use. TB cases with diabetes were known to have history of smoking with 73% (n=54) while non-smokers were 27% (n=20).
On sputum smear microscopy frequency of 3+ results showing high bacterial load, was profoundly higher i.e. 67.6% in diabetic tuberculosis patients as compared to non-diabetics which was 4.9% only. Total culture yield was 363 out of 500 sputum samples. There were 193 samples that were sensitive to all drugs, 9.4% were MDR strains (resistant to Isoniazid and Rifampicin). MDR-TB is significantly higher in TB-DM patients i.e. 13.5% as compared to 8.7% in TB only patients.
In our study, DNA sequence data for drug resistance was studied by the sequence of rpoB gene of the wild type MTB strain. Sequencing results showed mutations at various spots of rpoB gene.
Most common mutational sites identified were at codon 531, 526 and 516 with frequency of 70%, 15% and 7.5%, respectively. Moreover, mutation sites at 512 and 574 codon had also been reported. In this study, predominantly two phylogenetic variants were identified. Majority of the isolated strains were Central Asia Strain (CAS) with a prevalence of 88.2% and rest were Beijing strain. However, attempts to find zoonosis could not be established. A total of 900 raw milk samples were also screened for M. bovis and no positive sample could be detected.
The present study emphasizes the importance of screening for DM in TB patients, which had not been done in routine. This practice may prove to be helpful in reducing the disease burden of TB patients as well as DM patients. Thus it is recommended that the screening for DM should be implemented in TB/DOTS clinics.
Emergence of Multi drug resistant Mycobacterium tuberculosis is also a serious challenge for clinicians. A very large financial implication in terms of treatment, duration of chemotherapy and spread of MDR TB strains is being faced. Treating MDR TB is more complicated than treating drug sensitive TB. Patients with MDR TB require longer, much more costly treatment and experience higher mortality rates. Such a long time to initiate the treatment is not affordable, thus there is a dire need for some rapid technique like molecular based diagnostics for MDR detection, which can provide quick results and making it possible to start treatment at earlier to minimize transmission, morbidity and mortality.
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Epidemiology And Control Of Gastro-Intestinal Nematodes Of Large Ruminants In Balochistan
Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: The main area of research in this study was to assess the prevalence, hematological aspects
of Bovine nematodiasis. Three main experiments were conducted to highlight the objectives of
the present research study.
The first experiment was conducted to find out the prevalence of large ruminants major
nematodes for one year. For this purpose buffalo and cattle of either sexes and between < 1 year
to > 2 years of age were selected from two sites i.e., Quetta and Qilla Abdullah. Fecal analysis of
these cattle and buffalo showed overall higher (33.99%) nematodes prevalence recorded in buffalo
in Quetta, (27.99%) in cattle at Qilla Abdullah followed by in cattle at Quetta (26.66%). Five
nematode infection was recorded in all two experimental sites with higher prevalence of
Haemonchus contortus in buffalo at Quetta and Ostertgia ostertagi in cattle at Quetta and Qilla
Abdullah. The buffalo and cattle of < 1 year presented higher nematodes prevalence than 1-2 years
and > 2 years. The female buffalo and cattle were infected with nematodes prevalence higher
than male animals. These five nematodes were prevalent almost throughout the year, however a
peak infection was recorded during August and September in cattle and October in buffalo. The
high temperature, rainfall and humidity during these months may be predisposing factor of higher
prevalence. Mostly the level of nematodes infection was low(< 800 EPG) and did not seriously
impaired the buffalo and cattle productivity.
Second experiment on assessing the comparative efficacy of anthelmintics (Levamisole,
Oxafendazole and Ivermectin) against cattle and buffalo nematodes were conducted at Govt and
private farms. The results showed that Ivermectin than Oxfendazole were found effective against
cattle and buffalo nematodes. The higher (89-100%) reduction of EPG were recorded in cattle and
buffalo calves treated with Ivermectin followed by Oxfendazole (86-100%), Levmisole (88-
Third experiment was conducted to determine the hematological values in healthy and
nematodes infected animals. Different hematological parameters i.e., TEC, TLC, Hb estimation,
were determined. The results showed that overall low Hemoglobin estimation and RBC were
recorded in nematodes infected animals than healthy, while higher WBC were recorded in
nematodes infected animals than healthy. The Lymphocytes and Neutrophil and Monocytes were
higher in some nematodes and lower in other, while higher mean Eosinophil counts was recorded
in all nematodes infected animals than healthy animals.
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Effect Of Pre-Weaning Diets And Varying Levels Of Concentrate During Post-Weaning Period On The Performacne Of Female Nili-Ravi Buffalo Calves Up To One Year Of Age
Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: Nili-Ravi buffalo is a well-known buffalo breed in subcontinent Indo-Pakistan region and famous for its high milk production ability. Currently, buffalo calves and growing heifers are fed on deprived quality and quantity roughages with poor nutritive values resulting in reduced growth rate, reproduction with delayed attainment of puberty and high mortality. These constraints can be overcome through nutritional management of buffaloes. There is a need for the development of standards for adequate, cost effective provision of colostrum, whole milk/milk replacer and calf starter ration to neonatal calves up to weaning, establishment of nutrient requirements for growing buffalo heifer with aim of more average daily gain to reduce age at puberty and nutrients requirements for lactating buffalo according to their status and stage of milk production.
The current study comprises of two experiments and was conducted at Livestock Experiment Station, Bhunikey, Pattoki, District Kasur, Punjab, Pakistan. The first experiment was performed with an aim to check the growth performance of female buffalo calves on whole milk & milk replacer and find out the cost effective and growth rate friendly alternate source of liquid diet. The duration of this experiment was 120 days. Thirty six female calves were selected and randomly divided into three (n=12) different treatments A (whole milk), B (50% whole milk & 50% milk replacer) and C (milk replacer). All the calves were given colostrum for first three days, then whole milk up to 15 days of age and transferred into three treatments. In addition to this all the calves were provided calf starter and fresh water ad-libitum. The calves were given
liquid diet @ 10% of their body weight for first two months and then gradually decline of 1% on weekly basis for the subsequent two months. Green fodder was started on three month of age. The average daily total dry matter intake was remained same for all the three treatments but the average daily gain was higher in treatment A (457.38±110.13a) compare to treatment C (362.22±107.83b) but it was same for treatment A&B and B&C, respectively. The mean FCR value was also better for treatment A (3.49±0.56b) compare to treatment C (4.30±1.24a) and it was same for treatment A&B and treatment B&C, respectively. The mean cost/kg gain was higher in treatment A (422.72±70.66a) compare to treatment C (352.97±97.49b) and it was same for treatment A&B and B&C, respectively. Animals had performed well on mix liquid (50 % whole milk: 50% milk replacer) diet and it was more cost effective than other two treatments.
The aim in second experiment was to set the standard and cost effective level of concentrate ration for growing female buffalo heifer calves. For second experiment thirty (30) calves were selected from first experiment and were randomly dived into three treatments A, B and C. Treatment A was fed on concentrate ration according to 0.5 % of their body weight, treatment B 1.0 % and treatment C 1.5 % of their body weight. In addition to this all the calves were given ad-libitum green fodder and fresh clean water. All the calves were fed on similar concentrate ration having CP: 17 % and ME: 2.6 Mcal/kg. The duration of this experiment was 8 months. There was significant difference (P<0.05) in mean dry matter intake, protein intake, energy intake and protein per kg gain across all the three treatments and were higher (P<0.05) for treatment C then treatment B and lower (P<0.05) in treatment A, respectively. The average daily gain was remained same (P>0.05) for all the three treatments (497.32±17.92, 503.63±19.09 and 532.77±20.67). The higher feed efficiency was observed in treatment A (0.135±.004a) while it was same for treatment B & C (0.113±.003b & 0.108±.004b), respectively. The average body
condition & score, body mass index and blood constituents (RBCs, WBCs, heamoglobin, packed cell volume, mean corpuscular volume, platelets count, lymphocytes, monocytes and granulocytes) were unaffected (P>0.05) by different concentrate levels. Concentrate levels had significantly affected some of serum components (total protein and urea) but some components (glucose & cholesterol) were unaffected by dietary treatments. The values of mean serum total protein and serum urea were found lower in treatment A (6.12±0.17b & 42.34±1.59b) compare to treatment B (6.65±0.23a & 50.08±2.05a) and C (6.79±0.23a & 51.41±2.29a), respectively. The higher values of serum total protein and cholesterol in treatment B & C may be attributed to higher concentrate level in these two treatments. Concentrate levels had significantly (P<0.05) affected some of the digestibility parameters (DM %, CP% and NDF%) while other parameters (organic matter, fat, ash, ADF and urine pH) were remained same (P>0.05) on varying concentrate level diet. The mean body measurements (height at wither, body length and heart girth) were also not affected (P>0.05) by dietary treatments. There was significant difference across all the three treatments in total average daily dry matter intake cost and cost per kg gain. These were lower in treatment A compared to other two treatments B & C. It was observed that mean dry matter, protein and energy intake was lower in treatment A (0.5% of body weight) and weight gain was remained same on all the three dietary treatments. The mean feed efficiency was greater and mean cost per/kg gain was lower in treatment A. So, treatment A was remained more cost effective than other two treatments.
Both experiments were planned by keeping in mind the problems of buffalo farmer. Rearing of calves with improved growth rate on least cost feeding regime is important in dairy farming. Milk replacer is an alternate source of whole milk. Most of the buffalo farmers don’t use milk replacer for rearing of calves because of slower growth rate. Mixing of milk replacer
with whole milk in 50:50 ratio make the consistency of liquid diet near to whole milk. Feeding of whole milk with milk replacer along with calf starter reduces the cost without affecting growth rate. At this stage farmers should keep in mid the cleaning of feeding pans to avoid the risk of diarrhea.
In post weaning period calves’ rumen is fully develop and is completely shifted to solid diet. During this transition phase farmers don’t follow the nutritional requirements of calves, which slow down the growth rate and ultimately increase the age at puberty. As buffalo are efficient converter of low quality diet. If farmers offer concentrate ratio (16-18% CP) to buffalo heifers at the rate of 0.5% of body weight along with ad-libitum green fodder, growth rate can be improved cost effectively.
The findings of first experiment shows that 50% whole milk & 50% milk replacer @ of 10 of body weight along with adlibitum calf starter ration help in early rumen development, improved growth rate and better FCR on economical basis. So, it is recommended that whole milk and milk replacer in 50:50 ratio is growth rate friendly and cost effective for rearing of female buffalo calves up to weaning. The results of second experiment shows that growth rate, body measurements and body condition & score remained the same on all the three dietary concentrate levels but the feed efficiency was improved on lower concentrate level. So, it is recommended that it is cost effective to raise buffalo growing heifers on small amount of concentrate ration (0.5% of body weight) along with ad-libitum green fodder.
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