1.
Status Of Brucellosis And Its Effect On Hemogram And Serum Biochemistry In Indigenous, Cross-Bred And Exotic Dairy Cattle Herds
by Muhammad Hareem Afzal (2008-VA-250) | Dr. Muhammad Avais | Dr. Jawaria Ali Khan | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Brucellosis mainly infects food animals such as cattle, buffalo, goats and sheep. Brucella abortus is the principal cause of brucellosis in cattle and is shed from the infected animal at or around the time of calving or abortion. The present study was conducted on 450 animals on three different strains/breeds of cattle i.e. Exotic (150), Cross-bred (150) and local cattle (150) from 10 different privately owned livestock farms of varying holdings of district Lahore. An epidemiological questionnaire focusing on herd traits as well as husbandry and sanitary practices that could be associated with the risk of Brucellosis infection was completed. Serum samples were collected and analyzed using Rose Bengal Plate Test (RBPT). The serum samples positive for Brucellosis through RBPT further subjected to Serum Agglutination Test (SAT). To check the effect of Brucellosis on hemogram, blood samples from 18 cattle (n=6 indigenous; n=6 cross-bred; n=6 exotic) positive for Brucellosis and 18 animals (n=6 indigenous; n=6 cross-bred; n=6 exotic) negative for brucellosis were collected and processed for TLC, DLC, RBC, Hb, MCV, MCHC MCH and platelets using automated haematology analysed at UDL, UVAS, Lahore. Similarly, to see the effect of Brucellosis on Serum biochemistry, serum samples from 18 cattle (n=6 indigenous; n=6 cross-bred; n=6 exotic) positive for Brucellosis and 18 animals (n=6 indigenous; n=6 cross-bred; n=6 exotic) negative for brucellosis collected and analysed for glucose, total protein, albumin, Creatinine, Alanine Aminotransferase (ALT), Aspartate Aminotranferase (AST) and Sorbitol Dehydrogenase (SD) using commercially available kits.
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RBPT revealed overall prevalence 17.7% higher than SAT 10.6%. Prevalence of brucellosis is higher in Cross-Bred (22.7%) followed by local cattle (18.9%) and exotic (12%).
Hemato-boichemical results showed that increase in TLC, MCV While slight changes in Hb, MCHC, RBC and values of MCV stays within normal range. On the other hand serum biochemistry increase in AST while decrease in ALT and SD found. Availability: Items available for loan: UVAS Library [Call number: 2348-T] (1).
2.
Molecular Identification And Treatment Of Theileriosis In Small Ruminants Of Northern Balochistan
by Mir Ahmad Khan (2005-VA-214) | Prof. Dr. Muhammad Arif Khan | Prof. Dr. Muhammad Azam Kakar | Prof. Dr. Muhammad Sarwar Khan | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation 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
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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
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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. Availability: Items available for loan: UVAS Library [Call number: 2690-T] (1).
3.
Molecular Identification And Treatment Of Theileriosis In Small Ruminants Of Northern Balochistan
by Mir Ahmad Khan (2005-VA-214) | Prof. Dr. Muhammad Arif Khan | Prof. Dr. Muhammad Azam Kakar | Prof. Dr. Muhammad Sarwar Khan | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation 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
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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
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106
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. Availability: No items available
4.
Development And Evaluation Of Vaccines Prepared From Staphylococcus Aureus Isolates Of Camel Mastitis
by Amjad Islam Aqib (2013-VA-947) | Dr. Muhammad Ijaz | Dr. Riaz Hussain | Prof. Dr. Aneela Zameer Durrani | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Development And Evaluation Of Vaccines Prepared From Staphylococcus Aureus Isolates Of Camel Mastitis Availability: Items available for loan: UVAS Library [Call number: 2750-T] (1).
5.
Clinico-Epidemiological Study Of Multiple Drug Resistant Staphylococcus Aureus From Bovine Mastitis
by Muhammad Abdul Rauf Malik (2015-VA-832) | Dr. Muhammad Ijaz | Dr. Syed Saleem Ahmad | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Bovine mastitis is one of the most significant problems of livestock causing huge economic losses in dairy industry of Pakistan. Among other bacterial etiology of bovine mastitis, the Staphylococcus aureus is overwhelming to control and is well-known to cause subclinical and contagious mastitis. Methicillin resistant Staphylococcus aureus is our prevailing field issue. In view of the economic importance of Staphylococcus aureus mastitis, the current project was designed to study the Clinico-epidemiology of multiple drug resistant Staphylococcus aureus in bovine mastitis.
A total number of 900 milk samples (n=450 cattle, n=450 buffalo) were collected from Faisalabad district of Punjab, The collected samples were processed in laboratory of Microbiology and Medicine, Faculty of Veterinary Sciences, University of Veterinary and Animal Sciences, Lahore. Primarily, screening of subclinical mastitis was done by Surf Field Mastitis Test (SFMT). Milk samples were spread out primarily on blood agar to rule out possibility of loss of later growth. Colonies with round and golden color characteristic were put to sub-culturing on Mannitol salt agar as differential and selective medium. The morphological and microscopic clarification was done under microscope using Gram’s staining technique. Various biochemical tests including coagulase and catalase were applied.
Prevalence of subclinical mastitis was found 55% (495/900), however significant differences were found among different tehsils of district Faisalabad. The prevalence of subclinical mastitis in cattle from district Faisalabad was found 54% (243/450) that upon comparison between different cities presented significant difference. While buffaloes presented 56% of subclinical mastitis. Comparisons of subclinical mastitis among different tehsils were
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found significant.
Among quarter based prevalence of subclinical mastitis and prevalence of blocked quarters the number of blocked quarters were found 5.58% (201/3600) with highest percentage of blocked quarter noted in case of front right followed by rear right , front left , and rear left with 6.89, 6.56, 4.67, and 4.22%, respectively. However, the quarter based prevalence was found 32% (1088/3399) from bovine. The association of bovine subclinical mastitis with different risk factors presented significant association with few exceptions. Age, parity and body health status of animal were found non- significant with prevalence of mastitis. The breed and open rearing system presented significant relation with chances of mastitis. Chi-square test was used to statistically correlate the risk factors and prevalence of subclinical mastitis. P-value less than 0.05 was considered as significant.
Molecular confirmation of S. aureus was done by using coag gene through PCR technique. The S. aureus which were isolated from milk samples were put to antibiotic (oxacillin) sensitivity test for estimation of prevalence of methicillin resistant S. aureus from the bovine mastitis. Molecular identification of mec-A gene in staphylococcus aureus was done through PCR. The PCR confirmed methicillin resistant S. aureus isolates from cattle (n=20) and buffalo (n=20) were tested for their in-vitro drug response. However, Ciprofloxcin, Moxifloxacine, Linezolid, and Trimethoprim + Sulphamethoxazole were found 100% effective against multiple drug resistant Staphylococcus aureus and Levofloxacin showed 90% efficacy in bovine. While Oxytetracycline, Tylosin, Gentamycin, Amikacin, Vancomycin, and Fusidic acid were also found sensitive moderately except cefoxitin which was responsible for 100% resistane in bovine. Gentamycin was found to much more effective in buffaloes rather than cattle.
Summary
87
The study provided current status of Staphylococcus aureus infection with higher percentage in bovine mastitis. The prevalence of mecA gene revealed variation in methicillin resistant Staphylococcus aureus. In-vitro drug trial provided effective treatment possibilities against multiple drug resistance Staphylococcus aureus. Availability: Items available for loan: UVAS Library [Call number: 2815-T] (1).
6.
Development Of A Cost-Effective Serodiagnostic Assay For Peste Des Petits Ruminants (PPR)
by Tahira Hanif (2015-VA-1060) | Dr.Jawaria Ali Khan | Dr.Aamer Bin Zahur | Dr. Muhammad Avais | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Peste des petits ruminant (PPR) is a highly contagious newly developing disease of Small ruminants (sheep and goats). Currently the poor small ruminant’s farmers in Pakistan are facing huge economic losses due to PPR virus. In Pakistan PPR causes economic losses of Rs. 20.5 billion annually. The objectives of present study were to develop a cost effective sero-diagnostic assay for PPR (active haemagglutination inhibition and passive haemagglutination inhibition) and determination of comparative efficacy of active and passive haemagglutination inhibition assay (HI and PHA respectively) for detection of PPR virus infection.
In the present study, n= 300 sera samples were collected from sheep and goats during the (15 Februry 2016 to 2 January 2017).The serum samples were collected from kotli AJK :20(8 goats and 12 sheep),from gilgit:30 serum (20 goats and 10 sheep),from mansehra:22 serum (13 goats and 9 sheep),from mithi:112 (60 goats and 52 sheep) and 116 serum samples (88goats and 28 sheep) from Dhera ghazi khan.None of the animal was known to have been vaccinated against PPR previously or at the time of sampling.
These samples were collected from animals showed symptoms of PPR suggestive of PPR disease as well as from healthy animals. The sera were transferred into sterile tubes and were preserved on ice packs while shifting to the laboratory. PPR virus isolate was originally isolated from an outbreak in Taxila village, district Rawalpindi, the isolate was attenuated serially onto the Vero cell lines up to 20 passages. After, which antigen was titrated using a micotiter haemagglutination (HA) test with chicken RBCs and stored at -70◦c until use as a PPR antigen in a HI test.
In this study Active haemagglutination inhibition (HI) and passive haemagglutination inhibition were developed. The Haemagglutination Assay was standardized by different factors i.e. diluents, Temperature of incubation, Time of Incubation and concentration of Chicken R.B.C̓s. An additional test passive haemagglutination inhibition was performed to check the comparative efficacy of Active and Passive haemagglutination inhibition. In passive haemagglutination inhibition tanned sensitized cells remains effective due to their long effective life when stored at 4̊c and its makes an ideal test for diagnosis of PPR. Newly developed assays were compared against cELISA for PPR using kappa statistics and diagnostic sensitivity and specificity were determined.
The results of both assays were compared with results of competitive enzyme linked immunosorbent assay. In this study cELISA was considered as golden standard. The relative sensitivity and specificity of Active haemagglutination inhibition is 94.9% and 97.9% respectively. (Kappa 0.9264). However the sensitivity and specificity of Passive haemagglutination inhibition is 91.1% and 95.0% respectively.(kappa 0.8595). This study describes the serological detection of PPR virus by Active haemagglutination and passive haemagglutination inhibition (HI and PHA respectively). It was also concluded the comparative efficacy of (PHA and HI) that Active haemagglutination inhibition is more reliable technique than passive haemagglutination inhibition assay for the diagnosis of PPR disease in small ruminants (sheep and goats). Availability: Items available for loan: UVAS Library [Call number: 2816-T] (1).
7.
Prevalence, Molecular Diagnosis And Chemotherapy Of Degnala Disease In Large Ruminants Of Punjab.
by Mudassar Nazar (2005-VA-92) | Prof. Dr. Muhammad Sarwar Khan | Dr. Muhammad Ijaz | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: In Pakistan, Livestock is considered as a social security for poor villager as it can be a source of cash at the time of need. Degnala disease reduces the production of these animals directly. Along with other side issues related to Degnala disease, this study was done to diagnose the actual cause of Degnala disease by applying different latest scientific techniques. Prevalence along with risk factors was calculated in the rice growing areas of Punjab, Pakistan. Fungal isolation (n=40) was performed from the rice straw feedings of the Degnala disease affected animals through the technique of spot culture on SDA. Then these fungal isolates were identified through comparing their microscopic and macroscopic characters. Then toxigenic potential was checked for all these isolates through the application of TLC and HPLC. After that, from those isolates which were positive for mycotoxin production potential, most cytotoxic isolate was checked with the application of MTT assay. Then the most cytotoxic isolate was inoculated on non-contaminated rice straw and fed to the experimental animals to see a similarity of natural cases of Degnala disease. Finally treatment was conducted to see a proper combination of various drugs against this disease.
Toxigenic potential of different candidate fungi, isolated from rice straw feeding of Degnala disease affected bovines was analysed along with Species, age, gender and season wise prevalence. Out of 1536, 104 (6.77%) cases showed positive signs for this disease with a significant association (p<0.05) between rice straw feeding in buffaloes, winter season and bovines having an age of more than one year. Complete Blood Count showed marked increase in erythrocyte sedimentation rate and all white blood cells numbers, except lymphocytes in positive cases. There was a significant increase (p<0.05) in Alanine amino transferase, Aspartate amino transferase and Alkaline phosphatase noticed in Liver Function Test. At the same time, increased value of Creatinine was noticed in Renal Function Test. For isolation and screening of toxigenic fungi, rice straw samples (n=40) being fed to the positive cases were processed further, out of which there were 85 fungal isolates mainly of Aspergillus (57), Penicillium (10), Fusarium (04), Zygomycetes (03), Curvularia (01) and unidentified (10). All isolated fungi were subjected for mycotoxin production and only 11 showed mycotoxin producing capability (including Aspergillus, Penicillium and Fusarium isolates) analysed by Thin Layer Chromatography and quantified through High Performance Liquid Chromatography. It is concluded that all the fungi, contaminating rice straw feeding of Degnala affected animals are not toxigenic. This work will help in establishing major mycotoxin producing fungi leading to the probable cause of Degnala disease in bovine.
With the help of MTT assay on vero cell line, most cytotoxic fungus was identified. After an incubation with vero cells, OD values of all the candidate fungi were compared through one way ANOVA. Results of this analysis showed that Fusarium was at the highest ranking and then was the A. flavus with a significant value of 0.006 and 0.039.
Finally it was concluded through these systematic steps of converging the diagnosis that, out of all the 85 suspected fungi, Fusarium (isolate number S 8.1) was the most cytotoxic isolate obtained from the rice straw feedings of Degnala affected animals in our study.
For molecular diagnosis of the most cytotoxic isolate of Fusarium, PCR was conducted and the results showed that ultimately the final PCR product was successfully amplified against the mentioned primer of ITS conserved region for Fusarium genera and the DNA product was with a length of 570 base pairs.
Experimental feeding trials were conducted by inoculating Fusarium (the most cytotoxic isolate) and A. flavus (second most cytotoxic one after Fusarium) separately and in combination compared with the negative control group, all groups were of eight animals each. It was concluded that alone Fusarium was able to produce Degnala disease, while its combination with A. flavus was more lethal.
Ultimately the treatment trials proceeded with penta-sulphate, oxytetracycline and antiseptic topical application as therapeutic treatment were shown to be very effective against Degnala cases. While in all the affected animals feeding of affected rice straw was ceased. Only withdrawal of affected rice straw from the feedings of Degnala affected animals was not effective unless proper treatment as mentioned here was not conducted.
analysed The expected results of the study shall be helpful to make exact diagnosis and treatment of infected buffaloes and cattle that is further helpful for timely prophylaxis and control of the Degnala disease in the rice growing areas of Pakistan and South Asia.
Availability: Items available for loan: UVAS Library [Call number: 2960-T] (1).
