1.
Comparative Studies On The Sensitivity Of Polymerase Chain Reaction (Pcr) And Microscopic Examination For The Detection of Trypanosoma Evansi in Horses
by Muhammad Asif Muieed | Prof. Dr. Zafar Iqbal Chaudhary | Dr. Azhar Maqbool | Mr. Asim Aslam | Faculty of Veterinary Sciences.
Material type: ; Format:
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Publisher: 2005Dissertation note: The polymerase chain reaction (PCR) was standardized and its efficacy was evaluated against microscopic examination i.e. Giemsa stained smear method ['or the diagnosis of Trypanosoma evansi infection (Surra) in horses. l3lood samples were collected from 100 suspected horses from different localities in Lahore.
Under aseptic precautions blood smears were prepared, after drying and fixing with methanol, slides were stained by Giemsa stain method of staining. By stained blood smear method 5 out of 100 horses were found positive For T. evansi infection. The polymerase chain reaction (PCR) was carried out on the blood of' the same suspected horses to evaluate its efficacy in the diagnosis of' T. evansi infection and to compare its diagnostic value against the microscopic examination method currently in use. For this purpose total genomic DNA was extracted from suspected blood samples. The PCR reaction was performed in a 50tl reaction mixture containing I X Taq BuFfer, 0.2 mM dNTP Mixture. I .5 mM MgCIl2 2.5 U/1i1 Taq Polymerase. 4uM of' each primer, 2 ul of DNA extracted and 31.5 p1 of DNase - free deionised water. The tubes containing the mixture were subjected to 30 cycles of amplification in a thermocycler. During each cycle the sample of' DNA was denatured at 93° C' For 30 seconds, annealed at 45° C For 30 seconds and extended at 720 C For I minute. Prior to the cycling and at the end of' cycling the mixture was subjected to incubation at 93° C for a period of 3 minutes and final extension at 72° C for a period of 5 minutes, respectively. PCR product was then characterized by 2.5% of agarose gel electrophoresis. To confirm the presence of DNA and to estimate its size it was compared with a DNA ladder and was photographed with a Polaroid camera. The polymerase chain reaction (PCR) revealed 16 positive cases out of 100 above mentioned suspected cases. These 16 positive cases diagnosed by polymerase chain reaction (PCR) also included animals, which were diagnosed by stained blood smear method.
It can be concluded that polymerase chain reaction (PCR) is a superior and sensitive (16%) in comparison with the microscopic examination i.e. Giemsa stained smear method (5%). Polymerase chain reaction (PCR) is more effective in cases where the parasitemia is low and this test could be used in other species of animals especially camels where the disease is more chronic and difficult to confirm by. other routine methods. PCR would not only ensure early diagnosis and treatment in individual animals but can detect animal reservoirs of infection and would help to eliminate threat to equine and camel herds which are grazed and housed together and where blood sucking mechanical fly vectors are ever present.
Availability: Items available for loan: UVAS Library [Call number: 0860,T] (1).
2.
Detection Of Toxoplasma Gondii Infectionin Butchers And Buffaloes By Polymerase Chain Reaction (PCR) and Latex Agglutination Test
by Rana Sajjad Ahmed | Prof. Dr. Zafar Iqbal Chaudhary | Prof. Dr. Muhammad Naeem Khan | Faculty of Veterinary Sciences.
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Publisher: 2005Dissertation note: Toxoplasmosis, a common parasitic zoonotic infection is usually aymptomatic in immunocompetent persons although it may be present as lymphadenopathy, febrility, etc. but it is a life threatening opportunistic infection in congenitally infected patients and in immunocompromised individuals (those with AIDS, malignancy, organ transplantation, etc). Human beings become infected with T. gondii usually by ingesting oocysts in food and water contaminated with cat feces or by consuming tissue cysts in undercooked meat.
The diagnosis of toxoplasmosis is mainly based on serological tests latex agglutination test (LAT). Detection of specific DNA seems to be of clinical value in the ingestion of patients infected with toxoplasmosis.
In this study, latex agglutination test was used for the detection of the antibodies against Toxoplasma gondii and Polymerase Chain Reaction (PCR) based on the amplification of repetitive B1 gene of T. gondii. The study was based on a total of 200 samples involving 50 butchers, 50 buffalo's sera and whole blood respectively.
LAT established an overall infection of T. gondii in butchers and buffaloes as 20 % and 22 % respectively. The PCR analysis confirmed this T. gondii prevalence in butchers and buffaloes.
LAT proved to be an efficacious method for routine serological screening for antibodies to T. gondii. The costly and sophisticated PCR results in our investigation showed good correlation with the serological data of these patients showing that LAT can be used as an alternation to PCR. The results demonstrated that PCR analysis of clinical samples of patients suspective for acute toxoplasmosis including those with an acquired infection presented by lymphadenopathy can be a promising diagnostic method that enables direct detection of parasitic DNA.
Availability: Items available for loan: UVAS Library [Call number: 0861,T] (1).
3.
Diagnosis And Prevalence Of Trypanosoma Evansi In Camels Through Polymerase Chain Reaction (PCR) And Haematocrit Centrifugation Thechnique (HCT) in Punjab (Pakistan)
by Jahanzaib | Prof. Dr. Zafar Iqbal Chaudhary | Prof. Dr | Prof. Dr. H.A. Hashmi | Faculty of Veterinary Sciences.
Material type: ; Format:
print
Publisher: 2005Dissertation note: The most important protozoan disease of camels is trypanosomiasis caused by Trypanosoma evansi. There was little epidemiological information on the prevalence of infection. The present study was conducted to find out the prevalence of Trypanosorna evansi in camels through haematocrit centrifugation technique (HCT) and polymerase chain reaction (PCR). A total number of 100 camels of different age and sex groups were selected from different localities including Bahawalpur, Lahore, Gujranwala and faisalabad to find out the prevalence of Trypanosomiasis in Punjab (Pakistan) and to evaluate the sensitivity of PCR assay and HCT for the diagnosis Trypanosoma evansi. Blood samples were collected and examined by haematocrit centrifugation technique (HCT) and polymerase chain reaction (PCR). The prevalence was recorded as 4% and 13% by haematocrit centrifugation technique (HCT) and polymerase chain reaction. The positive samples by the polymerase chain reaction also included the positive animals by the haermatocrit centrifugation technique.
The results showed that PCR was more sensitive method for the detection of trypanosomiasis as compared to the haematocrit centrifugation technique. Thus PCR can be used for the diagnosis of camel trypanosornosis during both acute and chronic phases of infection, and for use in the evaluation of treatment. Application of PCR to field diagnosis is therefore clearly indicated.
Availability: Items available for loan: UVAS Library [Call number: 0862,T] (1).
4.
Molecular Diagnosis Of Bovine Tuberculosis In Humans
by Muhammad Bilal | Prof. Dr. Zafar Iqbal Chaudhary | Dr. Asim Aslam | Dr. Mnsur-uddin | Faculty of Veterinary Sciences.
Material type: ; Format:
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Publisher: 2005Dissertation note: Tuberculosis is a highly infectious disease. In humans it is mainly caused by Mycobacterium tuberculosis and occasionally by Mycobacterium bovis and Mycobacterium africanum. Bovine tuberculosis caused by M bovis is the main cause of enteric TB in humans. it is transmitted through milk, meat and dairy products. It is also recorded that it can also cause pulmonary tuberculosis in humans.
A study was conducted to detect the M bovis in human pulmonary sputum samples through PCR based techniques. A PCR assay was described which could differentiate M bovis from M. tuberculosis in clinical samples. Sputum of 400 patients was randomly analyzed with PCR assay. Two (0.5%) out of 400 sputum samples were positive for M bovis while remaining were positive for M tuberculosis. Over all 0.5% cases were positive for M bovis causing pulmonary tuberculosis in humans.
The two positive cases were analyzed in the background of their history. History revealed that both of them belong to different families and areas were in close contact with animals for a long time. It suggested that they caught infection from animals. It was an evidence of pulmonary tuberculosis of M bovis in humans.
Availability: Items available for loan: UVAS Library [Call number: 0865,T] (1).
5.
Standardization Of Tuberculin Test In Buffaloes And Detection Of Mycobacterium Bovis In Blood Through PCR
by Asad Ullah Khan | Prof. Dr. Zafar Iqbal Chaudhary | Dr. Asim Aslam | Prof. Dr. Abdul Rauf Shakoori | Faculty of Veterinary Sciences.
Material type: ; Format:
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Publisher: 2007Dissertation note: Tuberculosis is a highly infectious disease. In bovine it is mainly caused by Mycobacterium bovis. Bovine tuberculosis caused by M bovis is the main cause of enteric TB in humans. It is transmitted through milk, meat and dairy products. Bovine TB is still a significant zoonosis in many parts of the world and it accounts for 25.8% of TB in man.
A study was conducted to standardize the tuberculin test in buffaloes and to detect the M bovis in buffalo blood samples through PCR based techniques. A total of 100 buffaloes were tested by Single Comparative Cervical Intradermal Tuberculin Test (SCCIDTT) for this research and 100 blood samples were also collected from the same under aseptic condition. Data was also collected from owners & milkers of buffalo before and after SCCIDTT. A PCR (is a nucleic acid-based technique that enables the rapid and sensitive detection of micro-organism) assay was described which could detect M bovis in blood samples. Blood of 100 buffaloes was randomly analyzed with PCR assay. Over all two (2.0%) out of 100 buffaloes were found positive to tuberculin test while fifty four (54 %) out of 100 blood samples of the same buffaloes were found positive for M bovis in PCR.
The positive cases were analyzed in the background of their history. History revealed that the animals herd was crowded and were reared much closed to each other for a long time. It suggested that they got infection from other animals. It was an evidence of bovine tuberculosis of M bovis in buffaloes.
Availability: Items available for loan: UVAS Library [Call number: 0951,T] (1).
6.
Comparison Of Multiplex Pcr & Conventional Methods For The Diagnosis Of Tuber Culosis (TB) in Human, Buffalo & Cattle in Lahore District
by Naima Mumtaz | Prof. Dr. Zafar Iqbal Chaudhary | Dr. Asim Aslam | Prof. Dr. Abdul Rarf Shakoori | Faculty of Veterinary Sciences.
Material type: ; Format:
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; Literary form:
not fiction
Publisher: 2007Dissertation note: Tuberculosis, one of the most widespread infectious diseases, is the leading cause of death due to single infectious agent among humans and animals in the world. It is endemic in Pakistan with about 1.5 million people infected, and Pakistan ranks seventh among the 22 high-burden tuberculosis countries worldwide (WHO, 2006). Mycobacterium tuberculosis is the most common cause of human TB, but an unknown proportion of cases are due to Mycobacterium. bovis.
The study was conducted in Lahore to compare the multiplex PCR and conventional methods for the diagnosis of tuberculosis caused by M tuberculosis and M bovis in 300 humans' sputum and 1000 bovines' milk samples. Conventional methods included Ziehi Neelsen staining, culture and biochemical tests. For M tuberculosis and M bovis the pncA gene and specie -specific 500 bp fragments were targeted respectively. The sensitivity and specificity of multiplex PCR was found statistically significant in comparison to Ziehl Neelsen staining and culture for the differential diagnosis of TB. Pyrazinamide resistance was found in 15 (34.8%) out of 43 isolates recovered from media inoculated by sputum and milk.
Availability: Items available for loan: UVAS Library [Call number: 0954,T] (1).
7.
Comparative Studies On The Sensitivity Of Polymerase Chain Reaction (Pcr) And Conventional Serological Methods For the Diagnosis of Bovine Brucellqsis
by Raheela Akhtar | Prof. Dr. Zafar Iqbal Chaudhary | Prof. Dr. Abdul Rauf Shakoori | Prof. Dr. Asim | Faculty of Veterinary Sciences.
Material type: ; Format:
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; Literary form:
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Publisher: 2007Dissertation note: The polymerase chain reaction was standardized and its efficacy was evaluated against Rose Bengal Plate Test (RBPT) and Milk Ring test (MRT) for the diagnosis of brucellosis in 200 cows and buffaloes from Lahore and Okara districts of Punjab. Under aseptic measures 200 serum and 200 milk samples were tested by RBPT, MRT and PCR on both milk and serum samples in both cows and buffaloes as described in materials and methods. RBPT showed high sensitivity values (27.7% in cows and 45.2% in buffaloes) than serum PCR (25% in cows and 3 9.6% in buffaloes) but on other hands MRT showed low sensitivity (11.1% in cows, 25.4% in buffaloes) and high specificity (98.4% in cows and 93.6% in buffaloes) than milk PCR with sensitivity of 13.8% in cows, 29.4% in buffaloes and specificities of 95.2% in cows and 89.3% in buffaloes respectively. The comparison of PCR assays conducted on both types of samples showed high sensitivity of serum PCR against milk PCR. The comparison of RBPT and MRT in both species showed high sensitivity of RBPT than MRT. But due to low positive predictive value of RBPT and instability in its results in both species it is concluded that there is no significant difference in PCR and serological methods so no single test can be used for the exact diagnosis of bovine brucellosis.
Availability: Items available for loan: UVAS Library [Call number: 0957,T] (1).
8.
Diagnosis Of Bovine Tuber Culosis In Deers Kept In Captivity By Pcr And Tuberculin Test
by Zeeshan Nayyer | Prof. Dr. Zafar Iqbal Chaudhary | Dr. Asim Aslam | Dr. Azhar Maqbool | Faculty of Veterinary Sciences.
Material type: ; Format:
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Publisher: 2007Dissertation note: Tuberculosis is an infectious, chronic, granalomatous, highly communicable, zoonotic and debilitating disease. The etiological agents of tuberculosis belong to the bacteria Mycobacterium bovis. A total of 50 blood samples from emaciated deers were collected from deer’s kept in captivity suspected from TB. These samples were subjected to DNA extraction for polymerase chain reaction and tuberculin test for the sensitivity and specificity of these tests.The results obtained were analyzed by standardization of PCR for M. bovis.
PCR is a nucleic acid based technique that enables the rapid and sensitive detection of microorganism. Results indicated that 4% and 20% of deers were positive for M. bovis infection with the tuberculin test and polymerase chain reaction (PCR) respectively. From the results it is evident that polymerase chain reaction (PCR) technique is more sensitive than the tuberculin test for the diagnosis of tuberculosis and gives much higher percentage of positive cases.
Availability: Items available for loan: UVAS Library [Call number: 0970,T] (1).
9.
Epidemiology, Molecular Diagnosis And Chemotherapy Of Giardiasis In Bovine
by Sultan Ayaz | Prf.Dr. Azhar Maqbool | Prof. Dr. Zafar Iqbal Chaudhary | Faculty of Veterinary Sciences.
Material type: ; Format:
print
Publisher: 2009Dissertation note: Giardia is a protozoan parasite of the small intestine that causes extensive morbidity worldwide. Dairy calves can excrete high numbers of the cysts of Giardia and the disease in cattle is clinically important and can reduce the growth performance of the ruminants. Giardia is the cause of non-viral diarrhoea in humans and is responsible for epidemics in the developed and developing countries. The cyst is the infectious form, is ingested in contaminated water or food or directly from faecal-oral contact. Giardia duodenal is the only species, which is found in both humans and animals including dogs, cats, bovines, pigs, sheep and equine.
The present study was conducted to determine the prevalence in bovines at Military dairy farm, Gawala dairy colonies, the Government dairy farm and Household dairies in Lahore. The effect of season, sex, and age on infection rate and shedding of the cysts were also noted, and association of the Giardia infection with normal and abnormal stools was also studied.
Overall 2160 bovine faecal samples (720 buffaloes, 720 cattle and 720 calves) were examined during the study period from August 2007 to July 2008, amongst calves 362/720 (50.27%) were found to be positive. The highest prevalence was recorded in the Government. Dairy farm (68.33%) followed by Gawala colonies (55%), then the Military dairy farm (44.33%) and the lowest (34.44%) was recorded in Household dairies. Overall, highest (61.6%) seasonal prevalence was recorded during autumn, followed by spring (60.83%), then summer (53.4%) and the lowest
(34.1%) was recorded during winter. The highest (65%) prevalence was reported during August and the lowest (3 0%) during December.
Females were found to be more susceptible (56.74%) than males (35.1%). The prevalence was significantly higher (71.52%) in younger calves than the adults
(36.11%) (P<0.05).
Overall prevalence in cattle was 28.05%. The highest (41.67%) prevalence was recorded at the Government dairy farm, followed by Gawala colonies (32.72%), then the Military dairy farm (22.72%) and the lowest (15%) was recorded in Household dairies. The highest (35%) prevalence was found during August and the lowest (21%) during January. A significant difference (P<0.05) was noted. Females were found to be more susceptible (29.21%) than males (18.75%). The young calves had significantly higher (3 8.88%) prevalence as compared to the adults (24.44%).
Similarly, the overall prevalence in buffaloes was found to be 20.11% percent. The highest (40.55 %), prevalence was recorded at the Government Dairy Farm, followed by Gawala colonies (30%) then Military Dairy Farm (21.11%) and the lowest prevalence i.e. 12.77% was reported in Household Dairies. A non significant difference was recorded P>0.05). The highest (46.66 %) prevalence was recorded during August, while, the lowest (6.66%) during November and December. Females were found to be more susceptible than males. Where as the prevalence in a younger buffalo was significantly higher as compared to the adults.
Comparison of direct microscopic examination and PCR based methods was made at the Government dairy Farm, Gawala colonies; Military Dairy Farm and
Household Dairies. By direct Microscopic examination prevalence was found to be 28.05% (202/720) in cattle whereas by PCR it was 31.11%. Statistically analysis showed that the prevalence by PCR was significantly (P<0.05) higher than the microscopic examination.
It was observed that the highest prevalence of Giardiasis in bovines (Calves, Cattle and buffalo) was noted during August when the average temperature was 31.48°C. However the maximum and minimum temperatures were 35.37°C and
27.6°C, relative humidity 7 1.28% and rainfall was 3.2mm. The results of therapeutic trials by using albendazole, metronidazole, and mebendazole in cattle were calculated on the basis of reduction in the cysts count in the faeces after treatment.
Efficacy of albendazole at three dose levels i.e. 1 Omg/kg.b.wt, 1 5mg/kg.b.wt, 2Omg/kg.b.wt was 86.33%, 98.5% and 100% respectively, on day 27 after treatment. Efficacy of the metronidazole at 5Omg/kg.b.wt, 1 OOmg/kg.b.wt, and 1 5Omg/kg.b.wt. Was 85.42%, 87.8% and 94.02% respectively on day 27. Efficacy of mebendazole at three dosage level i.e. 7.5rng/kg.b.wt, lOmg/kg.b.wt and 2Omg/kg.b.wt was 81.15 %, 87.32%, and 90.4% on day 27 after treatment. Among these drugs, albendazole at
1 5mg/kg.body.weight was found to be most effective drug in the elimination Giardia infection. The significant (P<0.05) decrease in the CPG count after treatment in all the three groups and dose levels was noted. A significant difference (P<0.05) was observed in the level of leukocytes and of eosinophisl of infected cattle at day 06 and day 13 post inoculation. The leukocytes/lymphocytes count of Giardia infected cattle was 58.09%. Whereas, eosinophils constituted of leukocytes 9.69%. The total proteins of the sample were studied by sodium doedocyl sulphate polyacrylamide gel ELECTROPHORESIS (SDS PAGE). The result indicated that 8 diffeent molecular weight peptide badns were identified with size ranges from 20 to 70 KDa and common bands reported at 20, 24 and 35 K Da
Availability: Items available for loan: UVAS Library [Call number: 1146,T] (1).
10.
Seroepidemiology, Zoonotic Potential And Chemoprophylaxis Of Leishmaniasis In Dogs & Human In Pakistan
by Haroon Duraani | Prof. Dr. Azhar Maqbool | Prof. Dr. Zafar Iqbal Chaudhary.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2010Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1547,T] (1).
11.
Prevalence, Identification And Pathogenesis Of Clostridium Chauvoei In Cattle And Buffaloes In Punjab
by Muhammad Asif Idress | Prof. Dr. Zafar Iqbal Chaudhary | Prof. Dr. Muhammad Younus.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2012Dissertation 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).
12.
The Camel and Its Diseases
by Zafar Iqbal Chaudhary.
Edition: 1st ed.Material type: ; Literary form:
not fiction
Publisher: U.A.E Al Bayan Printing and Publishing; 2000Availability: Items available for loan: Pattoki Library [Call number: 636.2896 Zafar 14771 1st 2000 CMS] (1), UVAS Library [Call number: 636.29 Zafar 14672 1st 2000 CMS] (1).
13.
Identification And Molecular Characterization Of Mycobacterium Bovis And Mycobacterium Paratuberculosis In Wild Cats.
by Zianab Tariq (2010-VA-234) | Dr. Muhammad Yasin Tipu | Prof. Dr. Zafar Iqbal Chaudhary | Prof. Dr. Aftab Ahmed Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Cases of wildlife diseased with Mycobacterium species are existing in Pakistan and result in high morbidity and mortality. Vaccine is the only preventive measure, but in wildlife the vaccine administration is a strenuous job. In Pakistan vaccination practice is not up to the mark and vaccination schedules are not being followed. Mycobacterial diseases have gain popularity due to their zoonotic effect. Scat samples from Lahore Safari Zoo and Lahore Zoo were collected and properly labelled. Conventional PCR along with Touchdown PCR was done using universal primer sets of M. bovis and M. paratuberculosis. The amplicons were run on agarose gel and the bands were observed under Gel Doc system.
The objective of the study was to detect the currently prevailing Mycobacterium bovis and mycobacterium avium subspecies paratuberculosis in wild cats in Pakistan.
However the results obtained from different kinds of PCR were negative, showing that the wild cat population of Lahore Zoo Safari as well as Lahore Zoo were free from Mycobacterium bovis and mycobacterium avium subspecies paratuberculosis. Availability: Items available for loan: UVAS Library [Call number: 2845-T] (1).
