Clinicopathological Changes Induced By Heat Stress, Their Resolution By Minerals And Vitamin C Supplementation In Quails
Khurshid Anwar | Dr. Asim Aslam | Prof. Dr. Muhammad Younus Rana. Material type: Book ; Format:
; Literary form:
Publisher: 2011 Dissertation note: The present study was designed to overcome heat stress in Japanese quails through potassium chloride, sodium bicarbonate and vitamin C solution in calculated amount in water on thermo tolerance, histopathology and hematology of quails exposed to heat stress: This experimental trial was carried out at Avian Research and Training Centre (ARTC), UVAS Lahore and tests were performed at Department of Pathology UVAS, Lahore. There were three replicates in each treatment group and each replicate was of twenty quails, each group was comprised of 60 birds. The body weight of each the bird was. recorded on weekly basis. Blood samples were collected on the 21, 22, 23, 24, 29, 30 and 31 days of treatment from each group to evaluate the serum potassium and bicarbonate level in the blood. For hematological parameters the blood samples were collected on 22, 28 and 31 day of treatment and the vital organs for histopathology were collected after slaughtering 3 birds from each group. The hematological parameters were studied and the data was analyzed by two ways ANOVA. Group A quails revealed significantly higher weight gain than those of group B, but no significant difference was observed, when all groups were compared. Significantly less weight gain was revealed by the quails of group B, when compared to all other groups. Comparison between groups A, C, D, E, F and G was non significant. The highest FCR was exhibited by the birds of group A while the group B showed the poorest FCR. Better FCR was exhibited by group C, D, E, F and G. Serum samples were obtained from each group, for bicarbonate and potassium determination by spectrophotometric method. Group A, C, E, F and G exhibited a significantly higher serum potassium level than those of groups B and D. A significantly higher bicarbonate level was revealed in the serum of group A, D, E, F and G as compared to group B and C, on day 23 and onwards. But no significant difference was observed in serum of groups A, D, E, F and G. Blood hematology revealed no significant difference in red blood cells of groups A, C, D, E, F and G. Group B exhibited a significant lower values of red blood cells, packed cell volume, basophils, monocytes and showed a significant increase H/L ratio and eosinophils when compared to all other groups. Histopathological studied showed infiltration of heterophils, hyperemia, congestion of liver, heart and adrenal gland. It is concluded that, quails of group B (kept in high environmental temperature) revealed a decreased weight gain, poorest FCR, decreased serum potassium and bicarbonate level, decrease in hematocrit, monocytes and basophils and increased in eosinophils and H/L ratio. Supplementation of electrolytes and vitamin C (125 mg/L KC1, 75 mg/L NaHCO3 and vitamin C 62.5 mg/L) in water effect on heat stressed quails exhibited the better results in term of weight gain, serum electrolytes, blood profile and histology than those quails kept in heat stressed condition with no supplementation. From the present results it is concluded that 125 mg/L of KCI, 75 mg/L of NaHCO3 and 62.5 mg/L of vitamin C solution in water, alone or in combination may be used in quails to combat the effect of high ambient temperature and heat stress.
Availability: Items available for loan: UVAS Library [ Call number: 1226,T] (1).
Immuo-Pathological Response Of Pigeons To Challenge Infection Of Newcastle Disease Virus (Ndv)
Yasir Amin | Dr. Asim Aslam | Prof. Dr. Muhammad Younus Rana. Material type: Book ; Format:
; Literary form:
Publisher: 2011 Dissertation note: This study trial was designed to evaluate and compare the efficacy of two locally available live Newcastle disease vaccines (Medivac ND LaSota and VRI, Mukteshwar ND vaccine), also to compare two routes (Oral and Ocular) of vaccine administration in term of antibody titre and assessment of protection against field (chicken) isolated virulent Newcastle virus challenge (NDV) in pigeons. Study of clinical signs, gross and histopathological lesion in different organ of non-vaccinated and challenged birds was also the part of our present study. For this purpose one hundred and twenty pigeons were purchased from the local market and screened for Newcastle disease antibodies using Hemagglutination inhibition test. Healthy pigeons were randomly divided into six groups i.e. A, B, C, D, E and F, comprising 20 birds each. Group E and F were kept as positive and negative Control respectively. Group A and C were vaccinated with Medivac ND LaSota vaccine at day 7th and 21st of experiment through oral and ocular route. Similarly Group B and D were immunized with VRI (Mukteshwar) ND vaccine through oral and ocular route respectively. At 28th day of experiment all the groups except group F were challenged with velogenic field isolate of NDV at a dose rate of 0.1 ml through ocular route. Serum samples were collected at day 14, 21, 28, 35 and 42 of experiment for the determination of antibody titre. Post-infection clinical signs in control positive group were i.e. anorexia, dullness, depression, decreased feed intake, discharge from mouth, greenish diarrhea, nervous manifestations, leg and wing paralysis. Gross lesions on different organs were hemorrhages in trachea, proventriculus, spleenomegaly and greenish intestinal contents. Medivac ND LaSota vaccine produced higher immune response in term of antibody titre as compared to VRI (Mukteshwar) ND vaccine. It was also observed that ocular route irrespective to vaccine type produced significantly (P<0.05) higher immune response than oral route. Vaccine strains used in this study efficiently induced immune response through ocular route, suggesting that implementation of this vaccination programs in future may prevent ND outbreaks in pigeons, especially in racing pigeons, and may prevent NDV spread to other avian species, mainly poultry.
Availability: Items available for loan: UVAS Library [ Call number: 1228,T] (1).
Haematologial And Immunological Effects Of Naturally Occurring Probiotic (Yogurt) And Garlic Supplementation On Broiler Chicks Vaccinated Against Newcastle Disease
Muhammad Ishaq | Dr. Asim Aslam | Prof. Dr. Muhammad Younus Rana. Material type: Book ; Format:
; Literary form:
Publisher: 2011 Dissertation note: The present project was undertaken to study the various hematological. immunological and gross/histopathological parameters in ND vaccinated chicks with different supplementation of yogurt and garlic. For this purpose one hundred twenty-six day old broiler chicks were divided into three experimental groups A, B and C. each having 42 chicks per group. The group B and C \\ere Further subdivided into 3 groups yl, y2, y3 and gi, g2. g3 respectively comprising of 14 chicks per group having different supplementation of yogurt and garlic respectively, while group A was kept as a control and fed basal diet having no supplementation. Our result showed that yogurt augmented serological response and help in increasing HI antibody titer in which 200 gm yogurt showed immense potential in increasing HI titer until last day of experiment. All levels of Garlic group augmented serological response in term of antibody titer hut remain statistically insigni flcant in increasing HI titer.The heterophil population and I I/L ratio was also improved in both yogurt and garlic groups however 40 grn garlic fed group showed a better response in increasing hctcrophil population. TLC remains insignificant both at treatment and levels throughout the experiment. Body weight gain and FCR was also improved with yogurt and garlic supplementation however 200 gm yogurt showed a curvilinear response over the range of yogurt bd levels. Yogurt and garlic supplementation also showed improvement in the development of immune organ such as spleen, thymus and liver, their I listological examination revealed that an increase in supplementation of yogurt and garlic have no lethal effect upon morphological structure of these organs. In conclusion, yogurt as a probiotic and garlic as a growth promoter agent displayed a greater efficacy in increasing HI titer, heterophils population and improving productive performqnce of broilers in which 200 gm yougurt/kg diet group and 40gm garlic/kg diet showed an immense potential in improving above traits, so their use in broiler diet should be considered instead of using costly commercial probiotics and antibiotics.
Availability: Items available for loan: UVAS Library [ Call number: 1229,T] (1).
Detection Of Mycoplasma Synoviae By Pcr And Its Histopatholohical Studies In Poutry Breeder In District Abbottabad
Sajjad Ahmad | Dr. Muti- ur- Rehman Khan | Prof. Dr. Muhammad Younus Rana. Material type: Book ; Format:
; Literary form:
Publisher: 2011 Dissertation note: Poultry, an important sub-sector of livestock, has emerged a cheaper source of protein for human consumption. Mycoplasmas are the smallest known bacteria, 300-800 nm in diameter and are capable of replicating outside the cell. Mycoplasma synoviae is a member of the class Mollicutes, order Mycoplasmatales, family Mycoplasmataceae. Mycoplasma synoviae (MS) is considered economically to be most important pathogen. Mycoplasma synoviae infections occur in poultry worldwide, affecting poultry and causes diseases like respiratory distress, synovitis and arthritis. Mycoplasma is transmitted from infected to healthy birds both by horizontal and vertical routes. Horizontally disease is transmitted via infected and healthy carrier birds, hatchery, housing, equipments, feeding and during transportation.
To have an insight on pathogenesis and reliable diagnostic techniques, the present project was designed to know comparative sensitivity of rapid agglutination test and polymerase chain reaction for MS diagnosis and to study the gross lesion and histopathological changes in chicken joints produced by MS.
The birds showing clinical signs that included respiratory i.e. tracheal rales, conjunctivitis, coughing, sneezing, ocular and nasal discharge and infectious synovitis were selected for sample collection. Initially the collected sera samples were examined by Rapid Serum Agglutination test. RSA and PCR tests were used in order to confirm the pathogenic agent. RSA and PCR positive samples were further processed for histopathological study in order to identify the lesions in tissues produced by causative organism. In field visits it was observed that the suspected birds were with pale comb, mild to severe lameness, dull, depressed, ruffled feather, conjunctivitis, oculo-nasal discharge, tracheal rales and greenish or sulfur faeces. Birds hock joints, toe joints and paws pad were swelled. The infected birds were occasionally found with generalized infection. The infected birds complicated with other diseases of poultry such as Newcastle and infectious bronchitis causes infection airsacculitis.
Rapid serum agglutination test was conducted at 14 broiler breeder farms. The birds at a farm were showing respiratory and infectious synovitis signs and symptoms, suspected to Mycoplasma synoviae. The tests were performed at the spot. A total of 239 sera samples were examined out of which 63 (26.35%) sera samples were positive for MS. The clinical samples were identified and confirmed as Mycoplasma synoviae infection by PCR. The amplified PCR product was given about 211 bp size while PCR buffer was used as negative control. A total of 213 samples were subjected to PCR and 65 (30.52%) revealed PCR positive results for tracheal swabs, 28.16% (20 samples out 71) showed positive results. For tracheal and lung 33.38 % (24 out of 71) and 29.57% (21 out of 71 samples) were positive, respectively. The PCR test successfully amplified the DNA of MS clinical positive samples. Sixty five out of 213 Mycoplasma synoviae isolates were positive in MS specific PCR while the other 148 samples were negative. The sensitivity and specificity of molecular method Polymerase chain reaction was 100 percent.
For histopathological studies the samples of different organs including trachea, lungs, liver, hock joints (articular cartilage, piece of synovial membrane) and foot pad were further processed. The trachea was examined. There was epithelial degeneration, desquamation. congestion, haemorrhages and inflammatory cell infiltration. The lungs were examined and it was revealed that there was marked congestion, haemorrhages, necrosis and mononuclear cells infiltration. Liver showed infiltration of lymphocytes, plasma cells and macrophages. Articular cartilage showing chondrocytes degenration. Synovial membrane was thickened due to infiltration of lymphocytes and plasma cell. Foot pad showed hyperkaratosis and thickning of epidermis, acanthosis, degeneration of cartilage, infiltration of both mononuclear and plasma cell.
It is concluded from findings of present study that PCR is more appropriate technique than RSA for timely diagnosis of Mycoplasma synoviae. However combination of findings of both techniques may be utilized for accurate detection of Mycoplasma synoviae from broiler breeder in district Abbottabad.
Availability: Items available for loan: UVAS Library [ Call number: 1316,T] (1).
Prevalence, Identification And Pathogenesis Of Clostridium Chauvoei In Cattle And Buffaloes In Punjab
Muhammad Asif Idress | Prof. Dr. Zafar Iqbal Chaudhary | Prof. Dr. Muhammad Younus. Material type: Book ; Format:
; Literary form:
Publisher: 2012 Dissertation note: In the first phase of the project, the sampling of diseased animals presumably affected by Black quarter was carried out from six districts of Punjab belonging to three different zones. Around two hundred and fifty samples from each zone were collected and were subjected to bacterial culturing and isolation procedures followed by biochemical identification mechanism. The prevalence of Black quarter in Cattle and buffaloes were thus calculated for each district and zone. Highest prevalence of BQ in Zone II was observed (27.2%) for cattle while in case of Buffaloes highest prevalence (3.2%) was noted in Zone I. similarly higher Prevalence of BQ was noted in 1st quarter of year for Zone I followed by zone II and III while 2nd quarter of season was showing higher prevalence of BQ in zone II and III.
During 2nd phase of experiment tissue samples were inoculated in RCM and blood agar for the re-isolation of C. chauvoei, identified on the basis of colony characteristics and later on subjected to biochemical tests for the confirmation of the isolated organism. Then it was further confirmed through Polymerase chain Reaction for the identification of the causative agent i.e. C. Chauvoei on the basis of 16S rRNA gene sequence. Another set of primers corresponding to alpha toxin gene sequence of C. chauvoeui was also used which strengthened the belief that this strain of C. chauvoei possessed alpha toxin producing ability.
During third phase of project blood samples collected were subjected to hematological estimation for buffaloes and cattle having confirmed as BQ This study revealed significant effect on RBC's count and white blood cells count (P<0.05), while Differential leukocyte count were also showing significant different as compared to Non-infected (P< 0.05). Serum samples were tested for the change in levels of different enzymes. It was found that blood-glucose level and ALT levels were not significantly higher (P>0.05) when compared with control values, Values of AST, CPK and LDH were found significantly higher (P< 0.05) in all infected animals.
Histopathology of affected muscle tissues of both cattle and buffaloes was done to study microscopic changes in the muscle fibers and surrounding tissues. Lesions were somehow disappointing as compared to the magnitude of gross lesions. There were segmental degeneration, Zenker necrosis, discrete edema, occasional neutrophils and emphysema in affected muscle.
Finally, alpha toxin (hemolysin) in culture supernatant of RCM broth was titrated against 2% washed RBC's of cattle, buffalo, sheep, goat, chicken, rabbit and mice to study the hemolytic activity of the toxin. It was found that highest percentage of hemolysis was observed in mice followed by cattle, sheep, buffalo, chicken and rabbit respectively at 25°C. Higher the dilution of toxin, lower the extent of hemolysis. At 37°C variable results were obtained. It showed the biological activity of alpha toxin is also temperature dependant.
Availability: Items available for loan: UVAS Library [ Call number: 1664,T] (1).
Immuno Pathological Effects Of Neem (Azadirachta Indica) In Commercial Broiler Chickens
Zahid Jawad | Prof. Dr. Muhammad Younus | Dr. Muti-ur-Rehman | Prof. Dr. Azhar. Material type: Book ; Format:
; Literary form:
Publisher: 2012 Dissertation note: These experiments were conducted to study the effects of Azadirachta indica admixed in poultry feed on weight gain performance, haemtological values,immune modulations, and toxic effects in broiler chickens. A total number of 144 commercial broiler 1-day old chicks were reared in the experimental sheds of the Department of Pathology, University of Veterinary & Animal Sciences, Lahore, The birds were fed with balanced commercial feed and water ad libitum. The birds were divided into 3 groups; A, B and C having fourty eight chicks each. Birds of all groups were sub divided into four groups of each i.e. A1, A2, A3 and A4; B1, B2, B3 & B4 and C1, C2, C3 and C4, respectively. Each of the sub groups containd 12 birds. Sub groups A4, B4 and C4 were control group with no medication. The birds of groups A, B and C were fed with poultry feed containing dry powder of neem leaves @ 2 gm, 4 gm and 6gm per kg of feed respectively. The birds of groups A1, B1 and C1 were treated with the herb from day 0 to 42 of their life. The birds of groups A2, B2 and C2 were given the neem from day 14 to 42 of their life, whereas the birds of groups A3, B3 and C3 were treated with the herb from day 28 to 42 of their life. Difference between weekly weight gain in the birds of groups A1, B1 and C1 was non significant (P>0.05) however the difference between weight gain in the treated and control groups was significant (P<0.05). The birds treated with the herb from day 0 of their life showed more weight gain. There was no difference in the haematological indices between all of the treated groups and the control groups. The neem treated birds showed increased antibody titers against ND and IBD viruses as compared to control groups. The values of ALP and ASTshowed decreasing trend when the level of neem leaf meal was increased in the ration. Serum creatinine and serum uric acid values posed a slight declining trend in the neem fed birds. There was a decrease in serum cholesterol level in the neem treated bird groups, the higher the concentreation of the herb, the lower the cholesterol value. The organ body weight indices showed that there was no significant difference in liver, spleen and thymus weights among treated groups as well between treated and control groups. There was absence of prominent gross pathological lesions in liver, spleen, kidneys and thymus, however some treated groups showed mild hypertrophied liver and kidneys as did the organs of the birds in control groups. No histopathological changes except a few mild changes were observed in liver, spleen, kidneys and thymus in the birds of experimental groups.
Availability: Items available for loan: UVAS Library [ Call number: 1774,T] (1).
Toxicopathological Alterations And Tissue Residue Of Colistinsulphate (Polymixin E) In Broilers
Muhammad Aslam (2012-VA-809) | Dr. Muhammad Raza Hameed | Prof. Dr. Muhammad Younus | Dr. Syed Ehtisham ul Haque. Material type: Book ; Literary form:
Publisher: 2014 Dissertation note: Poultry industry is the 2nd largest industry in Pakistan playing important role to meet daily protein requirement with an investment of more than 200 billion rupees. It is contributing 6.1% and 10.8% share in total contribution of agriculture and livestock the national GDP, respectively. Total poultry meat production in the last year was 987 thousand tones (Economic survey of Pakistan 2013-2014). Because of high density and lack of biosecurity measures, outbreaks of infectious disease are more common and often occurs. Different antibiotics are used rational and irrational to treat and control these outbreaks. Colistin is among the most widely used antibiotic in intensive poultry farming, it is used to combat gram negative bacterial infections particularly for collibacillosis and gastrointestinal tract infections (Hussain and Khalil 2013).
Colistin belongs to the polymixins group of antibiotics. It is also known as Polymixin E. Polymixins are structurally related substances consisting of a cyclic peptide with a hydrophobic tail, mainly produced from gram positive nitrogen fixing bacteria Bacillus polymyxa. Polymixins are classified into six subtypes namely polymixin A to F and among these polymixin B and E are commonly used for the treatment of gram negative infections in animal and human beings. Colistin is mixture of two subtype’s colistin A (polymixin E1) and colistin B (polymixin E2). Commercially, it is available in two forms colistin sulfate and colistimethate sodium but colistin sulfate is more stable cationic water soluble salt (Landman et al. 2000) and is easily available in markets.
colistin sulphate has lipophilic and lipophobic(hydrophilic) properties, that makes it easier to interact on bacterial cell membrane by displacing counter ions in lipopolysaccharides, influencing Mg efflux that’s why it is effective against gram negative bacteria (Poole and Sheffield 2013). Polymixins also presents antipyretic activity by binding endotoxins, especially lipid A, (the active compound of lipopolysaccharide). Owing to this property it is extensively used to treat gram negative infections in livestock and poultry industries (Hanasawa et al. 1990).
The use of colistin was restricted in the past due to its toxic properties mainly in nephrotoxicity. There are reports available in the literature indicating that the parental use of colistin leads to the alterations of biochemical parameters in chicken i.e. increase in glutamic oxaloacetic transaminase, catalase activity, creatinine while, adversely affect total plasma proteins (Ibrahin et al. 2011). In ostriches it leads to the development of neurotoxic signs along with histologically odema formation in pericardium, intestinal serosa and heart was observed (Landman et al. 2000).
Toxic effects of colistin were also reported in humans including nephrotoxicity, increased serum creatinine during the treatment (Falagas et al. 2005). As bioavailability for oral route is minimal and it is used parentally to treat systemic infections. Parenteral route may cause toxopathological effects in birds. Other effects includes change in aspartate aminotransferase and alanine aminotransferase which indicate liver damage. Red blood cells, white blood cells, packed cell volume and hemoglobin estimation were also decreased (Saleemi et al. 2013). The incidence of nephrotoxicity is severe but less common in oral administration as polymixins absorption through gastrointestinal tract is very slow and limited, and cannot be detected in plasma concentrations at ordinary doses (Falagas and Kasiakou 2006). Instead of all these toxic effects, now a days it is frequently used for the treatment against gram negative bacterial infections (Vaara 2010).
The emergence of bacterial resistance has become an important public health hazard throughout the world). Due to irrational use of antimicrobial drugs against different diseases (Naqvi et al.2013. Colistin is one of the most widely used antibiotic in poultry industry against diseases like collibacillosis, salmonellosis and clostridial infections through drinking water, feed additives and parenteral route in developing countries like Pakistan (Tanweer et al.2013). Parenteral use of colistin sulphate may leads to the deposition of residues in different tissues. The presence of its residues in edible parts may lead to the toxicity as well as development of antibacterial resistance in human.
Keeping in view the potential toxic effects and other hazards relating to the extensive use of colistin in poultry birds, present study has been designed to investigate dose dependent patho-morphological alterations and tissue residue in broiler chickens.
1. To study the toxicopathological effects of parenteral administration of colistin in broilers.
2.Investigation of the tissue deposition of colistin as residues and its withdrawal period.
Availability: Items available for loan: UVAS Library [ Call number: 2221-T] (1).
Seroprevalence Of Brucellosis In Pregnant And Aborted Women Of Rural And Urban Areas In Three Selected Districts Of Punjab Pakistan
Shakeela Anjum (2006-VA-172) | Dr. Iahtasham khan | Prof. Dr. Abdul shakoor | Prof. Dr. Muhammad Younus. Material type: Book ; Literary form:
Publisher: 2015 Dissertation note: Brucellosis is an important zoonotic disease of animals and human. According to OIE
(Office International des Epizooties), it is the second most important zoonotic disease in the
world after rabies Brucellosis is more common in countries with Poorly standardized animal
and public health programme. Human Brucellosis caused by B. abortus, B. abortus are small,
non-motile, aerobic, facultative intracellular, Gram-negative cocobacilli.
The symptoms of human brucellosis are undulant fever, headache, weakness, body
pain, and sometimes endocarditis, orchitis, or arthritis may develop. Prolonged use of
combination of antibiotics and human vaccine is main control measure strategies for human
brucellosis. This disease is under investigated and hence people are ignorant of this insidious
problem. Serological surveillance is a good and cheap tool to diagnose this problem. There is
also need to know that which diagnostic antigen (Pakistani or French) is more sensitive and
specific from disease eradication and control point of view. Unfortunately annual incidence
of brucellosis in whole Pakistan is unknown but they are expected above 2 per 100,000
people. The hypothesis of present study was that brucellosis is considered to be endemic in
Pakistan but there is no data available about seroprevalence of brucellosis in pregnant,
aborted, rural and urban women with different risk factors in Pakistan.
In present study sero-screening of total 199 serum samples of pregnant and women
presenting with abortions from district Jhang, district Chinoit and district Faisalabad. Test
was performed on 199 samples which showed 40 (20%) positives by RBPT (Pakistan) and 42
(21%) positives by RBPT (France) (Table 4.4). While the indirect modified ELISA showing
5 (2.5%) positive samples. All sera tested positives (5) with modified i-ELIA were also tested
positives with another specific and highly expensive ELISA (Verion Serion) at OIE reference
Laboratory for brucellosis, Germany. Prevalence data was analyzed by chi square test
using SPSS version 20 Software (Apache License, USA) to find out correlation between risk
factors and brucellosis prevalence.
Risk factors such as consumption of raw milk, contact with animals, and symptoms
were significant. In our opinion, indirect modified ELISA is more sensitive than RBPT
(Pakistan) and RBPT (French). RBPT, (Pakistan) can be used for primary screening of
brucellosis cases because of cross reactivity present in RBPT antigen and confirmation must
be made with a more specific and sensitive serological test, such as B. abortus-specific
indirect modified ELISA. The results of present study showed that RBPT, Pakistan antigen
showed almost same sensitivity and specificity as that of RBPT (Pourquier, France) antigen.
Indirect modified ELISA is more specific as compared to RBPT (Pakistan) and RBPT
(French). It is highly suggestive to combine serodiagnostics test with molecular detection
including PCR and Real time PCR to increase the detection rate of brucellosis. Human
ELISA kit is extremely expensive that cannot be afforded in developing countries like
Pakistan to screen the human. PCR technique is highly reliable and less time consuming. It is
highly suggestive to conduct the study on human brucellosis on other districts of south
Punjab as it is believed that brucellosis is endemic due to lack of awareness and vaccination
in these areas.
Availability: Items available for loan: UVAS Library [ Call number: 2359-T] (1).
Seroprevalence Of Camel Brucellosis In Three Selected Districts Of Punjab, Pakistan
Sana Fatima (2007-VA-455) | Dr. Iahtasham Khan | Dr. Amar Nasir | Prof. Dr. Muhammad Younus. Material type: Book ; Literary form:
Publisher: 2015 Dissertation note: This is a representative study from Pakistan conducted in three districts of Punjab i.e.,
Jhang, Chiniot and Bhakkar which are rich in livestock population. There is a lot of research
work on livestock and dairy animals but camels are neglected and under-investigated animals.
Nomads mainly depend upon camels for milk, meat, milk byproducts and their earnings. Indeed,
it is a precious animal for them and therefore, to ensure the good health of their animals it is
essential that owners maintain healthy husbandry conditions. Brucellosis is one of the major and
unreported problems amongst the camels of our country. It results in losses to the economics of
the farmers/owners in terms of poor health, abortions, long calving interval, production of weak
offspring’s and poor quality of milk and meat. This disease is under-investigated and hence
people are ignorant of this insidious problem. Serological surveillance is a good and cheaper tool
to diagnose this problem. Determination of the seroprevalence is important to know the load of
disease and pockets of infections in the areas of central Punjab, Pakistan. There is also a need to
know which diagnostic antigen is more sensitive and specific from disease eradication and
control point of view.
A total of 200 camel serum samples were collected from three districts of the Punjab
province and tested by conventional screening test i.e., Rose Bengal Plate Agglutination Test
(RBPT) by using two antigens of different countries origin i.e. RBPT (IDEXX, Pourquier,
France) and RBPT (VRI, Pakistan) and then tested with confirmatory competitive Enzymelinked
Immunosorbent assay (cELISA). The data thus obtained regarding seroprevalence was
analyzed by using Chi-square and logistic regression IBM SPSS Statistics 20 (Apache software
Of the total 200 camels (50 male and 150 female), 5% (10 of 200), 4% (8 of 200) were
seropositive for anti-Brucella antibodies. Different risk factors were also included in study like
origin (nomadic and organized), area, age, gender, season, type of herd, abortion history and
orchitis in male. All of these risk factors were statistically analyzed to reveal the truth about
camel brucellosis. Of the various risk factors studied, the risk factors including camels (cows)
positive history of abortion (45.5%), orchitis (camel bulls) (33.3%), rearing with other ruminants
(9.4%), winter season (11.5%), nomadic production system (2.67-6.67%), and area Jhang (6-
12%) and Chiniot (2-8%) were statistically significant which could be potential source of threat
for humans and other animals. The testing was performed using conventional methods as well as
using cELISA. It is concluded that cELISA is more specific than conventional screening tests but
molecular diagnosis is highly suggestive for future studies. ELISA performed in Pakistan
(cELISA, Svanovir) and the ELISA (iELISA, ID VET Kit, France) performed in OIE reference
lab for brucellosis in Germany showed perfect agreement between them (both tested four camel
sera positive). The current study will help to minimize and eradicate the low prevalence of camel
brucellosis by creating awareness amongst the farmers and through vaccination and herd
immunization of all camel calves at age of 4-8 months. Adopting the policy of testing and culling
of positive reactors will be pivotal to achieve the objectives.
Availability: Items available for loan: UVAS Library [ Call number: 2361-T] (1).
Seroprevalence Of Brucellosis In Ruminants At Sale And Purchase Markets And Abattoirs In Three Selected Districts Of Punjab, Pakistan
Muhammad Tahir (2008-VA-301) | Dr. Iahtasham Khan | Prof. Dr. Muhammad Younus | Dr. Syed Ehtisham-ul-Haque. Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: This is a representative study from Pakistan accompanied in three districts of
Punjab i.e. Jhang, Layyah and Toba Tek Singh which are rich in livestock population.
There is a lot of research work on livestock farms but the sale and purchase markets and
abattoirs are neglected. Pakistan is an agricultural country and livestock contribute a major
share to agriculture in Pakistan GDP and its economy. Sheep, goats, cattle and buffaloes
contribute in the economy of Pakistani people by milk and meat production on majority
basis. Livestock contribution to the agriculture value added is 56.3%, while it contributes
11.9% to the national GDP during 2014-15. More than 8.0 million rural families of the
country are involved in raising livestock. But productivity of livestock is questioned by a
lot of factors. Infectious zoonotic diseases are among these.
Brucellosis is one of the major and unreported problem amongst livestock markets
and abattoirs of our country. It results in losses to the economics of the farmers or owners
in terms of poor health, abortions, long calving interval, production of weak offsprings and
deprived quality of milk and meat. This disease is under investigated and hence people are
unaware of this deceptive problem. Serological surveillance is a good and cheaper tool to
diagnose this problem. Determination of the seroprevalence is important to know the load
of disease and pockets of infections in the area of Punjab, Pakistan. There is also a need to
know which diagnostic antigen is more sensitive and specific from disease eradication and
control point of view.
A total of 300 animals serum samples were collected from three districts of the
Punjab province and tested by conventional screening test i.e. Rose Bengal Plate
Agglutination Test (RBPT) by using two antigens of different countries origin i.e. VRI
(Lahore, Pakistan), IDEXX (Pourquier, France) and ID.vet, France and then tested with
indirect Enzyme-linked Immunosorbent Assay (iELISA). The data thus obtained regarding
seroprevalence was analyzed using Chi-square SPSS version -19 (Nahar et al. 2009).
Of the total 300 animals (77 male and 223 female), 1.3% male and 35.66% female
were seropositive for anti-Brucella antibodies. Different risk factors were also included in
study like sex, area, age, gender, abortion, reproductive problems and also history of orchitis
in male. All of these risk factors were statistically analyzed to reveal the truth about ruminants
brucellosis. Among the various risk factors studied, the risk factors including history of
abortion (16.44%), orchitis (1.3%), retained placenta (26.47%), species cattle (35.55%) and in
Toba Tek Singh (47.36%) were statistically significant which could be potential source of
threat for humans and other animals. The testing was performed using conventional methods
as well as using iELISA. It is concluded that iELISA is more sensitive and specific than
conventional screening tests but molecular diagnosis is highly recommended for future
studies. The current study will help to minimize and eradicate the low prevalence of ruminants
brucellosis by creating awareness between the farmers and through vaccination and herd
immunization. By implementing the policy of testing and culling of positive reactors will be
pivotal to achieve the objectives.
For confirmatory diagnosis all the sera were further tested with iELISA. For this
purpose an exclusive commercially available iELISA Kit (ID.vet, France) was used which
would be able to differentiate anti-Brucella antibodies of B. abortus and B. melitensis and this
iELISA kit would be able to be used for multispecies including cattle, buffaloes, sheep and
Availability: Items available for loan: UVAS Library [ Call number: 2556-T] (1).
Clinico-Pathological And Pathomorphological Studies On Co-Infection Of Avian Influenza (H9n2) With Escherichia Coli In Broiler Chicken
Shahid Jaleel (2003-VA-93) | Dr. Muhammad Asif Idrees | Prof. Dr. Muhammad Younus | Dr. Muhammad Arshad. Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: E.coli is an important pathogen of domestic poultry and is prevalent in commercial poultry. LPAIV H9N2 infections are emerging respiratory problems in poultry industry, causing huge economic losses especially in the presence of other co-infecting pathogens such as E.coli. Co-infections do occur but are not easily detected, and the impact of co-infections on pathobiology is unknown. The mixed infections may provide increased virulence, posing a substantial risk to poultry and public health. Moreover, mixed infections of low pathogenic avian influenza with bacteria can also lead to devastating pandemics and a major threat to poultry health, worldwide in future. Co-infections do occur but are not easily detected, and the impact of co-infections on pathobiology is unknown. The mixed infections may provide increased virulence, posing a substantial risk to poultry and public health. Moreover, mixed infections of low pathogenic avian influenza with bacteria can also lead to devastating pandemics and a major threat to poultry health, worldwide in future. The aim of the present study was to investigate the infection of LPAIV A/chicken/Pakistan/10RS3039-284-48/2010 (H9N2) in chickens challenged with E.coli (O78:K80).
This study had three objectives. First, it is designed to develop co-infection experimental models LPAIV (H9N2) + bacteria (E.coli) in the avian model. Second, it aims to study the hematological and biochemical alterations during co-infection in avian model. Finally to study the pathological and histological alterations during co-infection in avian model, this study will help researchers and veterinarians in implementation of necessary control measures. E.coli stockculture was prepared by inoculating MacConkey’s agar with a loop full of reference E.coli strain culture and incubating at 37°C for 24 h. The estimated colony count was confirmed by plating 0.1 ml of a 104 and a 105 dilution of the final culture onto separate MacConkey’s agar plates. Avian influenza A virus, A/chicken/Pakistan/10RS3039-284-48/2010 (H9N2) was obtained from Poultry Research Institute (PRI) Rawalpindi Pakistan.
Viral stocks were prepared and titrated in 9-day-old to 10-day-old chicken embryonated eggs the median embryo infectious dose (EID50) was computed using previously reported approaches The viral stocks were diluted in medium containing antimicrobials to give a final titre of 106 EID50/ ml The study were ran on 80 broiler chicks (3week old), procured from local hatchery. All fowl were held serologically innocent and free from flu virus by haemagglutination inhibition (HI).
Chicken were infected under experimental conditions with E.coli (O78:K80) and low pathogenic avian influenza (LPAI) strain (A/chicken/Pakistan/UDL-01/08) (H9N2) alone or in combination. The experimental groups were identified as follows: negative control, E.coli, AI, and E.coli plus AI. Infected birds showed clinical signs of differing severity, with the most prominent disease signs appearing in birds of the E.coli plus AI group. Moreover, birds in E.coli plus AI group showed significant decrease in weight, enhanced macroscopic and microscopic pathological lesions. Specifically, the survival rate was 60%, 90%, and 100% in birds inoculated with E.coli + AI, E.coli and control negative or AI virus alone, respectively. Hematological studies revealed anemia, thrombocytopenia and leukopenia especially in co-infected birds. Biochemical studies revealed a significant decrease in total protein, glucose and albumin concentration with significant increase of activities of aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase. Prominent increase in creatinine, urea and uric acid were significantly detected in the infected chicken. The results showed that experimental co-infection of E.coli and H9N2 increased the severity of clinical signs, mortality rate and gross lesions and suggest than E.coli infection can induce higher economic losses and mortality if H9N2 LPAIV is also present. The HI titer against LPAIV infection in the co-infected group was significantly higher than the HI titer of AI group, which may indicate that E.coli could promote the propagation of H9N2 LPAIV or stimulate the immune response. The present study revealed that co-infection E.coli and H9N2 LPAIV caused more serious synergistic pathogenic effects and indicates the role of both pathogens as complicating factors in poultry infections.
Availability: Items available for loan: UVAS Library [ Call number: 2552-T] (1).
Immunomodulatory Effect Of Hing (Ferula Foetida) Along With Infectious Bursal Disease (Ibd) Vaccine In Broiler
Muhammad Javed Raza | Prof. Dr. Muhammad Younus. Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: A number of feed additives including antibiotics have been extensively used in
poultry feeds. However, the use of antibiotics has been restricted due to the drug resistance
and the issue of residues in meat. Now a day, the use of medicinal plants is being popular as
an alternate remedy. Asafoetida is a natural feed additive and antimicrobial, immune
stimulator, antiviral, antifungal, anti-parasitic, antithrombotic, antioxidant, anti-cancerous,
and vasodilator activities. It is being used for the control of some viral problems. Some
previous studies also indicated that it has beneficial effects on the immune system. Therefore,
the present study was designed to estimate the immunomodulatory effects of F. foetida with
IBD vaccine in enhancing the immune system and ultimately increase the production of
A total of n=90 day old broiler chicks were purchased and kept under the optimum
conditions at CVAS Jhang. Birds were divided into three groups A, B, C and group B and C
were further divided into three subgroups (B1, B2, B3 and C1, C2, C3). Group A was treated
as positive control group and was administered with commercially available IBD and ND
vaccine and routine diet while group B was administered with asafoetida at the rate of 2%,
4%, and 6% along with vaccine to see the impact of different levels of asafoetida on the
immune system and to see the effects (if any) of various doses of asafoetida. Group C was
only administered with asafoetida in feed at the same rate as to group B. At the end of study
birds were slaughtered to observe the effects of asafoetida administration.
Positive effect of asafoetida has been reported by many studies. F. foetida is a
medicinal herb used for the prevention and treatment of many diseases, because of having
antiviral, antibacterial and antifungal activities. Also act as a good growth promoting agent
and have beneficial effects on the immune system. So, it was hypothesized that the present
study was helpful to determine the immunomodulatory effect of asafoetida for commercially
available IBD vaccine and could reduce the cost of vaccination for commercial poultry
Availability: Items available for loan: UVAS Library [ Call number: 2562-T] (1).
Immunomodulatory Effects Of Feeding Allium Sativum Against Infectious Bursal Disease (IBD) Vaccinated Broiler Birds On IBD Vaccine.
Uzma Riaz (2008-VA-286) | Prof. Dr. Muhammad Younus | Dr. Muhammad Asif Idrees | Dr. Iahtasham Khan. Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: A number of feed additives including antibiotics have been extensively used in
poultry diets for the purpose of weight gain to improve feed efficiency and growth rate.
However, use of antibiotics has restricted due to the bacterial resistance and the issue of
residues which make the chicken meat harmful for human consumption. So the medicinal
plants are gaining interest as alternative feed strategies now a day because of their low cost,
easy availability and presence of no residues. Garlic was used for the medicinal purposes and
as a health supplement by the ancient Egyptians. It is a natural feed additive and is
antimicrobial, immune stimulator, antiviral, antifungal, anti-parasitic, antithrombotic,
antioxidant, anti-cancerous, and vasodilator activities. Previous studies indicate that it has
beneficial effects on the immune system and is a best immune stimulator then the other
herbal plants and medicines.
Therefore, the present study was designed to estimate the immunomodulatory effect
of garlic to commercially available IBD vaccine in enhancing the immune system. Total
N=99 day old broiler chicks were purchased and kept in the experimental shed of CVAS
Jhang. Birds were divided into three groups A, B, C and group B and C were further divided
into three subgroups (B1, B2, B3 and C1, C2, C3). Group A was treated as control group and
was administered with commercially available IBD and ND vaccine and routine diet while
group B was administered with garlic at the rate of 4%, 5%, and 7% along with vaccine to see
the impact of different levels of garlic (Allium sativum) on the immune system and to see the
toxic effect (if any) of high dose of garlic. Group C was only administered with garlic in fee
at the same rate as to group B. At the end of study birds were slaughtered to check the effects
of garlic administration. Positive effect of garlic has been reported by many studies. Garlic is
a medicinal herb used for the prevention and treatment of many diseases, because of having
antiviral, antibacterial and antifungal activities. Also act as a good growth promoting agent
and have beneficial effects on the immune system. Results of the study indicate that
administration of garlic powder in different doses alone and combined with commercial IBD
vaccine have good effects on the growth, blood parameters and the immune system of the
Availability: Items available for loan: UVAS Library [ Call number: 2561-T] (1).
Seroprevalence Investigations And Molecular Detection Of Mycoplasma Synoviae Using Loop-Mediated Isothermal Amplification (Lamp) In Poultry
Madiha Kiran (2008-VA-272) | Dr. Syed Ehtisham ul Haq | Dr. Usman Waheed | Prof. Dr. Muhammad Younus. Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: Pakistan poultry industry has been making remarkable progress from the last few decades but it is still threatened to certain infectious diseases which may cause heavy economic losses. Respiratory distress is of major concern, caused by a number of bacteria and viruses. Mycoplasma synoviae is the second most economically significant bacterial pathogen causing infectious sinusitis which contributes greatest in respiratory distress cases, and infectious synovitis. Serology is a best tool to detect the prevalence of infection in a flock but sometimes it is associated with imperfections in results. Accurate and early diagnosis through molecular technique is necessary for timely treatment of the disease. Loop-mediated isothermal amplification (LAMP) represents a rapid and sensitive alternative to conventional PCR, showing to be a robust, inexpensive, simple, and powerful method for selective and specific detection of M. synoviae.
A total of n=300 sera samples were collected from broiler and layer flocks. Samples were tested against MS antibodies by Rapid Serum Agglutination (RSA) using SPAFAS MS Plate antigen (Charles River Inc., CT, USA). For LAMP based identification, tracheal and synovial swabs samples were collected from sero-positive birds. To perform LAMP, DNA was extracted through organic extraction method from swab samples, as well as from Indicating FTA® classic cards (for positive control). The vlhA gene targeted primers were designed using Primer Explorer V5. Primers were synthesized over 200 nmol scale. FIP and BIP were subjected to Reverse Phase Cartridge (RPC) for purification. The LAMP reaction mixture was incubated at 60оC temperature in a water bath for 60 minutes. After amplification of DNA, Bsm DNA
polymerase had been inactivated at 80оC. Results were read after gel electrophoresis in 2% agarose gel. A real-time LAMP assay was also performed using ESEQuant tube scanner with FAM as reporting dye.
The total samples positive for MS infection were 153 when tested through RSA, indicating 51% seroprevalence of MS in poultry flocks of Jhang. The positive percentage for broiler and layer birds was 48% and 54%, respectively. LAMP assay detected 54% positive samples. The results of successful LAMP assay indicated a specific ladder-like pattern. The developed LAMP assay was found 100% specific, as it did not amplify the DNA of other mycoplasma species (M. gallisepticum and M. imitans). The sensitivity of the Real-time LAMP assay was noted as 10fg/μl of DNA.
The study concluded that Mycoplasma synoviae infection is prevalent in poultry flocks of District Jhang of Punjab, Pakistan, and the LAMP assay is a sensitive rapid molecular method for the detection of M. synoviae.
In summary, the developed LAMP assay is an ideal, robust and simple method to diagnose M. synoviae in poultry. Based on its high specificity and sensitivity, LAMP can be considered as a promising tool for small as well as large-scale detection of M. synoviae in poultry flocks.
Availability: Items available for loan: UVAS Library [ Call number: 2589-T] (1).
Effects Of Aqueous Extract Of Neem (Azadirachta Indica) Leaves Against Colibacillosis In Broiler
Muhammad Azfar (2014-VA-1111) | Dr. Asif Idrees | Prof. Dr. Muhammad Younus | Dr. Iahtasham Khan. Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: In Pakistan the poultry sector has developed rapidly in the last two decades. The poultry production as practiced today is specialized one and concentrating more on the use of high performance birds. The major factors for successful poultry production are high genetic potential, balanced nutrition and health maintenance (Nayaka et al. 2012). Utilization of immunostimulant is one solution to improve the immunity of animals and to decrease their susceptibility to infectious diseases. Immunostimulation comprises a prophylactic and therapeutic concept aimed at stimulation of the non-specific and specific immune response (Hyde and Patnode, 2001).
Most of the commercial poultry growers use antibiotics as growth promoters and to reduce the chance of occurrence of infectious diseases, which usually result in higher costs of production and ultimately lower net returns. There are great concerns about the use of antibiotics as therapeutic immunomodulators and growth promoters as it has given rise to antibiotic resistant bacteria. In this modern world of science, the researchers are once again intending towards exploring the potential benefits of the conventional medicines. Neem (A. indica) is an herbal plant which is known to exhibit various beneficial pharmacological properties including immunomodulatory effect in broilers.
Poultry sector is facing some problems one of which is bacterial disease which cause economic loss. Farmers use different antibiotics as growth promoter and to reduce the incidence of different diseases on poultry which increases cost of production and lowers the net returns. In addition, use of antibiotics giving rise to other problems like increasing drug resistant against these antibiotics especially Gram –ve bacteria (Salmonella and E. coli). It’s necessary that antibiotic residues in meat would not adversely affect human health. Specifically, it has been recommended that the penicillins, tetracyclines, tylosin, and
sulfonamides as growth promoters should not be use any more. So scientists are giving attention to the use of indigenous medicine instead of antibiotics for better rearing of poultry. Pakistan is blessed with different medicinal plants one of which is Neem (Azadirachta indica). For thousands of years the medicinal properties of Neem have been recognized. Each part of the Neem tree has some medicinal property. Biological Activity of Neem is due presence of different chemicals like nimbolide, nimbin, nimbindin, quercetin among others. Biological activities include Anti-inflammatory; Antiarthritic; Antipyretic; Hypoglycemic; Antigastric ulcer; Spermicidal; Antifungal; Antibacterial; Diuretic; Antimalarial; Antitumour; Immunomodulatory A. indica grow fast and is evergreen tree which has medicinal and nutritive value for broilers. Broilers which are given Neem leaves extract in water have shown improved FCR and weight gain. This medicinal Plants compete with synthetic drugs as it strengthens the immune system by activate Cell mediated immunity and have no residues in meat. Neem extracts also improve performance and hematological parameters
However, I did the research work in short-term basis (only 42 days) and modern equipments were also not available. Therefore, an extensive research and development work should be undertaken on Neem and its products to make the findings more accurate and effective for their better economic and therapeutic utilization.
Availability: Items available for loan: UVAS Library [ Call number: 2613-T] (1).
Seroprevalence Of Brucellosis In Dairy Cattle In Three Selected Districts Of Punjab, Pakistan
Ahmad Raza (2008-VA-271) | Dr. Iahtasham Khan | Prof. Dr. Muhammad Younus | Dr. Muhammad Faiz Qamar. Material type: Book ; Literary form:
Publisher: 2016 Dissertation note: Brucellosis is an infectious disease of domestic and wild animals with serious
zoonotic implications in humans. The disease in animals causes tremendous economic losses.
Since brucellosis is primarily an animal disease, emphasis should be given to control the
disease in animal populations by adopting well organized control strategies to protect the
public from the risks of acquiring this zoonosis.
A total of 300 blood samples (N=300) were collected randomly from cattle species
with one hundred samples (n=100) from each district of Chiniot, Sargodha and Sahiwal.
Approximately 04 mL of blood was collected and transferred in a blood Vacutainer having
gel containing clot activating factor, stored immediately at 4 °C in an ice box and was
transported to laboratory. The serum samples were screened by RBPT which is screening test
for brucellosis and it was observed that 38 animals were seropositive by RBPT. Then serum
samples were further confirmed by the use of most specific and sensitive serological test
known as ELISA.
Out of 300 samples, 33 animals were confirmed as seropositive by Indirect ELISA.
The overall prevalence in each of this district by RBPT was found to be 12%, 12% and 14 %
in Chiniot, Sargodha and Sahiwal respectively. The overall prevalence in each of this district
by iELISA was found to be 10%, 11% and 12 %.
By RBPT, Seroprevalence in females was 13.57% based on gender, animals having
abortion history had high seroprevalence for brucellosis (30%) than the non-aborted animals
(10.83%) and the animals with history of retained placenta are 19.30%. Seroprevalence of
iELISA based on gender showed 11.79%, cattle having history of abortion 30% and animals having history of retained placenta in cattle (24.57%). The risk factors i.e. sex, abortion,
retained placenta and areas are highly significant.
Availability: Items available for loan: UVAS Library [ Call number: 2602-T] (1).
Pathogenesis Of Aflatoxin B1 In Quails Under Experimental Conditions And Detoxification By Biological And Chemical Means
Sakhra Mahmood (2005-VA-251) | Prof. Dr. Muhammad Younus Rana | Prof. Dr. Asim Aslam | Prof. Dr. Aftab Ahmed Anjum. 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.
Availability: Items available for loan: UVAS Library [ Call number: 2670-T] (1).
Effect Of Bacillus Subtilis And Sodium Butyrate On The Morphometry Of The Small Intestine And Immune System In Healthy And Salmonella-Challenged Broiler Chickens
Arbab Sikandar (2005-VA-154) | Dr. Hafsa Zaneb | Prof. Dr. muhammad Younus | Dr. Sima Masood | Prof. Dr. Asim Aslam. Material type: Book ; Literary form:
Publisher: 2017 Dissertation note: Supplementation ofBacillus subtilis and microencapsulated sodium butyrate in the feed is being practiced as a substitute for antibiotics growth promoters. An expansive range of encouraging health-related properties exhibited by B. subtilis and SB has been published, but their exact effect on gut and immune system is not completely understood. Consequently, the evaluation of B. subtilis andSB as feed supplements is desired. To achieve this goal, the present study was aimed to investigate the effects of B. subtilis and SB on performance, immune system, gut and lymphoid organs microarchitecture in healthy and Salmonella-challenged broiler chickens.
In the first experiment the research was targeted to investigate the effects of B. subtilis on performance, immune system, gut and lymphoid organ microarchitecture in broilers. A total of 120 d-old broiler chicks were randomly distributed into four groups, each group with three replicates containing 10 birds per replicate. The birds were fed a corn-soy-based basal diet (BD, control) or BD supplemented with 10% zinc bacitracin (ZnB), and 0.05g/kg or 0.1g/kg of B. subtilis, respectively. On d 21 and 35, six birds from each group were killed to collect blood and visceral organs (thymus, spleen, bursa of Fabricius, liver and small intestine). Parameters evaluated included growth performance, immune responses, relative organ weights, lymphoid organs and gut mucosal morphometry, intraepithelial lymphocytes (IEL) count and goblet cell histochemistry in mucosa. Results showed that the group fed 0.1g/kg of B. subtilis had superior (P<0.05) mean body weight and weight gain, and lower FCR compared to the non-supplemented or ZnB-fed groups.The BS-0.1 group revealed higher antibody titer against Newcastle disease (ND) virus and the supplemented groups against sheep RBCs (SRBCs) on d 35. Cell-mediated immune response post-phytohemagglutinin-P injection was attained (P<0.05) by birds in the BS-0.1 group at 24h, and by both the BS-0.1 and BS-0.05 groups at 48 and 72h compared to the ZnB and control groups. The BS-0.1 group gained higher (P<0.05) relative bursal weight on d 21 compared to the other groups. Compared to the control group, the liver, spleen and thymus weighed more (P<0.05) in the experimental groups on d 35. The histomorphological study revealed increased (P<0.05) thymus cortical width, and cortex/medulla ratio in the BS-0.1 group compared to the control. The area of the bursal follicles and germinal centers of the spleen also improved (P<0.05) in the BS-0.1 group compared to the control. Compared to the ZnB and control, higher (P<0.05) villus height, villus surface area and villus crypt ratio of the duodenum and jejunum were recorded on d 21, and higher (P<0.05) villus heightof the duodenum and ileum was noted on d 35 in the BS-0.1 and BS-0.05 groups. The number of goblet cells having acid mucin was significantly higher in the ileal mucosae of the BS-0.1 group chickens compared to the ZnB and control. In conclusion, B. subtilis type probiotics effectuated better growth performance, improved immune system and modulated morphology of lymphoid organs and gut mucosa in broilers.
The second experiment was carried out to evaluate the effects of sodium butyrate on growth performance, immune status, organ weights and the microarchitecture of lymphoid organs and the small intestine compared to the effects brought about by an antibiotic. The cell-mediated immune response at 48 h post-phytohemagglutinin-P injection, and antibody titer against NDV and sheep RBCs on d 35 was higher (P < 0.05) in SB-1 chicks compared to those in the ZnB and control groups. Higher (P < 0.05) weight gain, and lower (P < 0.05) FCR were attained by the supplemented groups compared to the control. The thymus and spleen weighed more (P < 0.05) in the SB-1 group and bursa registered more (P < 0.05) weight in both SB groups compared to the control. On d 21, areas of the thymus medulla and the spleen germinal centers were larger (P < 0.05) in SB-1 chicks compared to ZnB and control chicks. The VH and VSA increased (P < 0.05) in the duodenum and jejunum in both SB groups on d 21, and in SB-1 on d 35 compared to the ZnB and control groups. The villus to crypt ratio was higher (P < 0.05) in the duodenum in SB-1 chicks compared to ZnB and control chicks. On d 35, VH in all segments and VSA in the duodenum and jejunum increased (P < 0.05) in SB-1 chicks compared to ZnB and control chicks. Statistically, IEL count was not significant among supplemented groups. On d 21, the number of goblet cells containing acidic mucin increased (P < 0.05) in all the segments of the small intestines in the SB-1 group compared to the control group and on d 35 in the ileum compared to the other groups. In conclusion sodium butyrate elicited better growth performance, improved immune system and modulated the morphology of lymphoid organs and the gut mucosa in broiler chickens.
The third experiment was focused to assess the effect of B. subtilis and SB on gut development, growth performance and immune system in broilers challenged with S. Gallinarum. Better growth performance was reported in the supplemented groups compared to the NC-S group due to better feed efficiency. The B. subtilis-supplemented group exhibited higher (P < 0.05) cellular immunity and antibody titer against NDV compared to the PC-S and NC-S groups. Furthermore, B. subtilis¬- and SB-supplemented groups reflected higher (P < 0.05) relative thymus and bursa weights, and improved microarchitecture of the lymphoid organs compared to the NC-S group. On d 21, villus surface area in the jejunum and ileum increased (P < 0.05) in sodium butyrate-treated birds. The crypt depth of the jejunum decreased (P < 0.05) in B. subtilis and sodium butyrate groups compared to NC-S and PC-S groups. On d 35, the villus height, villus surface area and VH:CD ratio of the duodenum increased (P < 0.05) in the supplemented groups compared to the NC-S group. The FCR, Salmonella population in ceca and mortality were higher (P < 0.05) in the NC-S group. In conclusion, the prophylactic use of the B. subtilis probiotic and SB alleviated stress associated with SalmonellaGallinarum infection and improved performance, immune function, lymphoid organs and gut mucosal development in infected broilers. Further analyses are needed to reveal the mechanism(s) by which B. subtilis and sodium butyrate produce such effects.
Availability: Items available for loan: UVAS Library [ Call number: 2790-T] (1).
Molecular Detection & Isolation Of Benzimidazole Resistant Haemonchus Contortus And Evaluation Of Anthelmintic Activity Of Herbal Extracts
Kazmi Ali (2009-VA-392) | Prof. Dr. Muhammad FaizwQamar | Prof. Dr. Muhammad Younus | Dr. Iahtasham Khan. Material type: Book ; Literary form:
Publisher: 2017 Dissertation note: Recently, DNA-based techniques have been applied for specific identification and characterization of parasites and also to evaluate the level of infections. The most commonly used group of anthelmintics is Benzimidazoles (BZ). BZ includes Thiabendazole, Mebendazole, Albendazole, Oxfendazole, Oxibendazole, Febantel and Netobimin. It binds to β- tubulin and inhibits polymerization of tubulin dimers to microtubules H. contortus involves mutations in β-tubulin genes at specific amino acid sites determine the BZ resistance. This study involves the Haemnochus contortus (barber pole worm) present in abomasums of small ruminants. n= 300 total samples were randomly be collected from the different sampling locations. Out of 300 samples of worms, 30 worms had been subjected to DNA extraction and PCR reaction. PCR recipe was prepared with following composition with total volume of 20μl, Master mix (enzynomics) 10μl, Distilled water 4μl, Forward primer 2μl, Reverse primer 2μl and genomic DNA 2μl. After PCR product has been obtained, gel extraction was done by PureLink Quick Gel Extraction Kit of INVITROGEN for purifying the DNA to allow it for gene sequencing. Only selected positive amplification products had been subjected to DNA gene sequencing and were sent to Philco company for ABI type sequencing. The polymorphic sites had been identified by using the standard set of ambiguity nucleotides through BLAST searches at NCBI/primers used to generate the β- tubulin gene sequence from H. contortus. DNA nucleotide sequences obtained are aligned on BIO EDIT SEQUENCE ALIGNMENT EDITOR software to understand the mutation, deletion, replacement of nucleotides. Genetic variation exists in all the samples sent for gene sequencing. These findings show that genetic diversity and genetic variation existing in β-tubulin isotype 1 of the H. contortus population of small ruminants in Jhang and Toba Tek Singh districts of Central Punjab, Pakistan. Samples exhibits different gene sequence and are from Jhang and Toba Tek Singh district. Genetic sequences are retrieved with the sequence and research papers of previously work done on GENBANK NCBI shows 100% query cover and ident with the isolates and clone of China, UK, USA, Australia, Brazil and somewhat similarity of 94% query cover and 100% ident with isolates of India. Some of those H. contortus worms which have been identified as BZ resistant strains were subjected in vitro anthelmintic activity of the herbal extract. Herbal formulation manifests the anthelmintic activity in dose-dependent manner in AMA and EHT. Mortality of worms was comparable with the reference drug used Levamisole at 200mg/ml concentration just from the start of exposure in AMA. Conclusion drawn from the experiment is that the herbal extract efficacy is comparable with the efficacy of the synthetic anthelmintics and can be marketed as these are field friendly, environment safely, economically and socially efficient. However before starts commercialization and marketing of such products at large scale, these materials must be subjected to standardized protocols and procedures of parasitology to be accomplished.
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Pathological Effects Of Natural And Experimental Lead (Pb) Toxicity In Lohi Sheep At Jhang, Pakistan
Muhammad Sajid (2010VA-61) | Prof. Dr. Muhammad Younus | Dr. Muti ur Rehman Khan | Prof. Dr. Aftab Ahmed Anjum. Material type: Book ; Literary form:
Publisher: 2017 Dissertation note: Heavy metal toxicity is increasing due to increasing trends of urbanization and
industrialization. Lead poisoning has been recognized as a major public health risk,
particularly in developing countries. It is nephrotoxic, hepatotoxic, neurotoxic, carcinogenic
and mutagenic for animals and human. Sewerage water, fertilizers, leaded-gasoline and lead
based batteries are the sources of lead contamination in soil and forage. The lead particles are
taken up by animals from contaminated forages and excreted in animal products like milk and
The presence of Pb in drinking water, waste water, plant products and animal
products has been studied which is a serious risk for animal and public health. The
legislations for the disposal of household wastes and industrial effluents are very poor in
Pakistan. The calculation of safe Pb levels in different products is still to be needed.
Pathological effects of higher Pb levels have not been studied in Pakistan. The present study
was aimed to unveil the toxic effects at constant dose of Pb over a period of three months in a
local sheep breed of Pakistan. The status of Pb toxicity was also investigated in a polluted
area around sewage drain and mutton slaughter house at District Jhang, Pakistan.
The Pb concentration in soil, forage and irrigating water was found to be below the
permissible limits and was safe for agriculture but long-term ingestion of low Pb
concentration may have cumulative effect. The serum Pb concentration was found to be
above the recommended safe limits for producing Pb toxicity in animals. The different tissues
like kidney, liver and skeletal muscles also contained higher Pb level from the permissible
limits and found to be unsafe for public use. Kidney showed the highest Pb concentration and
the muscle contained the least Pb level in the present study.
The erythrocyte count, hemoglobin concentration and packed cell volume showed
inverse correlation with Pb concentration and mean values were below the normal range in
Pb treated sheep but anemia was not developed. The erythrocyte sedimentation rate was also
influenced by given dose of lead acetate during third month of treatment. The white blood
cells also revealed no effect on given dose of lead acetate in Lohi sheep in this study.
The biochemical parameters of field and treatment group showed higher concentration
as compared to control group of Lohi sheep but their means were falling within the normal
range of reference values. The disturbed biochemical parameters in apparently healthy sheep
with higher serum Pb concentration were indicative for liver and kidney damage.
Lohi sheep exhibited less effect on given dose of lead acetate during first two months
but more pronounced changes of chronic Pb toxicity were observed during last month of trial.
The histological changes were not observed on early period in lead acetated treated sheep.
The characteristic histological changes were observed on last slaughtering at day 90 in kidney
and liver including degeneration and focal areas of necrosis, dilatation of blood vessels with
accumulation of red blood cells and fibrosis in some areas. The nuclear changes were more
typical with intranuclear inclusion bodies in renal tubular epithelial cells but less
distinguishable in hepatocytes.
It was concluded that soil, forage and water contained low Pb levels in the study area.
The ingestion of low Pb level for longer period had cumulative effect in animals. The animals
might be resistant to low Pb level but their products are a severe risk for public health. So the
necessary measures should be adopted to minimize the heavy metal contamination in animal
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