1.
Pcr-Based Diagnosis Of Canine Parvovirus In Dogs
by Farhan Towakal | Prof.Dr.Masoos Rabbani | Prof.Dr.Khushi Muhammad | Prof.Dr.Zafar | Faculty of Veterinary Sciences.
Material type: ; Format:
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Publisher: 2007Dissertation note: Canine parvovirosis, caused by a haemagglutinating canine parvovirus (CPV), one of the most important acute viral infectious diseases of pups, had been prevalent in the country. In the present study, 40 faecal samples, from clinically suspected cases of parvovirus diseased dogs and stray dogs were collected. The samples were diluted and centrifuged to collect the supernatant. Being a haemagglutinating (HA) virus; the pre-filtered supernatant from all suspected samples was checked for any HA activity using 1% washed chicken erythrocytes. Out of total of 40 samples, 30 samples were found HA positive. All the HA positive samples and samples found negative were processed for extraction of viral DNA with Genomic DNA purification kit. The net isolated DNA samples were subjected to Polymerase Chain Reaction (PCR) by using specific primers (developed against the variable genomic region of the capsid protein of the DNA) of the CPV. One of the selected primer pair designated as (37, 38) amplified the genornic region present in the CPV-2, the other primer pair designated as (39, 40) that amplified the region present in the CPV-2b.Optimization of PCR was done for the molecular level diagnosis of canine parvovirus and this technique was previously not available or standardized in local conditions of Pakistan. With the use of primer pair (37, 38) best results were found on following optimizing conditions like annealing temperature 55°C, primers concentration (reverse and forward) 25pmoles, Magnesium Chloride concentration 2mM, DNA(Template) volume Spl, Taq DNA polymerase(12.5U) and 2001iM of each dNTPs. For the primer pair (39,40) best results were found on following optimizing conditions like annealing temperature 5 5°C, Magnesium Chloride concentration 1.5mM, primers concentration (reverse and forwid) 20pmoles, DNA(Template) volume 6il, Taq DNA polymerase(12.5U), 2001.iM of each dNTPs. The PCR products were analyzed and compared for their banding pattern of 681bp and 427bp amplified by the primer pairs (37, 38) and (39, 40), respectively, against the standard DNA ladder (1 OObp) by using horizontal agarose gel electrophoresis. One of the HJA positive samples was not amplified by the PCR. The results of this study showed that the specificity of PCR reaction as compared to the HA test and the presence of the CPV-2 and 2b in Pakistan as the prevalent pathogenic strains. This study has been a first step for the molecular characterizationldiagnosis of canine parvovirus in local conditions of Pakistan. It is hoped that this study will pave the way for further advanced studies on this topic.
Availability: Items available for loan: UVAS Library [Call number: 0972,T] (1).
2.
Comparative Efficacy Of The Transverse Pylorpplasty And Y-U Advancement Pyloroplasty As A Relief To Pyloric Stenosis in Dogs
by Ayesha Safdar Ch | Dr.Muhammad Arif Khan | Dr.Asim Khalid Mehmood | Prof.Dr..Zafar | Faculty of Veterinary Sciences.
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Publisher: 2007Dissertation note: Stomach is important and biggest dilatation of alimentary canal which mainly serves the purpose of digestion of food. The stomach has two extremities cardia and pylorus, it also has fundus and body. The pyloric extremity is connected with the duodenum by pyloric canal. The gastric emptying of food is controlled by pyloric sphincter. Many problem have been diagnosed in the pylorus such as gastric and duodenal ulcers, neoplaisa, chronic gastritis, drugs and chemicals resulting in pyloric stenosis. Various surgical attempts were made in the past to treat such problems, these includes pyloromyotomy, pyloroplasty, transverse and YU advancement pyloroplasty and partial or complete pylorectomy.
Keeping in view the importance of this part of stomach two surgical procedures were tried Transverse pyloroplasty and YU advancement pyloroplasty as a solution to the obstructive problems of the pylorus. Although these techniques are well established methods and have been used by the various surgeons in the past as a remedy to the stenotic problems of the pylorus but no work has been done on comparative efficacy of these two procedures concerned.
The present project was designed to study the comparative efficacy of these two techniques. Twelve healthy mongrel dogs were selected and were divided into three groups comprising four animals each.ln group A Transverse Pyloroplasty , in group B YU Advancement Pyloroplasty was performed and dogs of C group were kept as control. The efficacy of the procedures was evaluated on basis of physical examination, radiographic evaluation, and postmortem examination.
Laparotomy was performed in aseptic condition and pylorus was exteriorized in both the groups. In dogs of group A, longitudinal incision was given over the pylorus which was closed in transversed fashion. After checking any leakage at the operated site the abdomen was closed in routine manner. In group B, Y shaped incision was given at the pylorus which was converted into U. Dogs of group C were kept as control and they were not subjected to any surgical procedure.
Barium study procedure was performed in the dogs of all groups to estimate the rate of gastric emptying and post mortem was preformed to evaluate the change in diameter of the pylorus.
The results of this study clearly indicated that although both the techniques are effective in increasing the diameter and improving the gastric emptying but Y U Advancement Pyloroplasty was more useful and effective procedure.
Availability: Items available for loan: UVAS Library [Call number: 0988,T] (1).
3.
Diagnosis Of Surra In Equines By Indirection Fluorescent Anitobody
by Malik Ahsan Nadeem | Dr.Asim Aslam | Dr.Kamran | Prof.Dr.Zafar Iqbal Ch | Faculty of Veterinary Sciences.
Material type: ; Format:
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; Nature of contents: ; Literary form: Publisher: 2007Dissertation note: Trypanosomiasis (‘surra) is the most widely distributed arthropod- born protozoan disease affecting the equines. This study was conduàted to check the efficacy of indirect fluorescent antibody test (IFAT) for the diagnosis of Surra. For this purpose 200 blood samples were collected from horses and donkeys from different areas of Gujranwala district. Thin blood smears were prepared on clean glass slides and blood samples were centrifuged to separate the serum. Serum was transferred into the vacutainers and transported to laboratory. The serum was separated by centrifugation and stored at -70°C. 200 thin blood smear slides were fixed with methanol and subjected to Giemsa stain for further microscopic examination. Then the 200 thin blood smear slides were fixed with acetone for further processing in indirect fluorescent antibody test (IFAT). The prevalence rate of 2% and 6% by using thin blood smear and indirect fluorescent antibody test (IFAT) was obtained respectively. The results helped us to determine accuracy of indirect fluorescent antibody test (IFAT) for diagnosis of Surra.
Availability: Items available for loan: UVAS Library [Call number: 0994,T] (1).
4.
Study On Molecular Diagnosis Of Canine Distemper Virus
by Muhammad Zubair Shabbir | Prof.Dr.Masood Rabbani | Prof.Dr.Khushi Muhammad | Prof.dr.Zafar | Faculty of Veterinary Sciences.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2008Dissertation note: Samples from fourty five dogs were submitted to the University diagnostic Laboratory, University of Veterinary and Animal Sciences, Lahore from January, 2007 to January 2008 for diagnosis of CDV infection. These dogs presented to referring veterinarians with clinical signs suspicious of CDV infection. Hematological examination (lymphocyte count) was carried out using K-EDTA anti-coagulant added whole blood and RT-PCR tests were performed using biological fluid samples that include plasma, nasal and conjunctival swabs. Only distemper positive dogs by RT-PCR were followed up for subsequent lymphocyte count and prognosis of distemper infection.
All the distemper positive dogs were lymphopenic but the degree of severity was variable as the samples were collected from dogs of different ages and phase of the disease. The study revealed that lymphopenia can be used to support presumptive clinical diagnosis but required laboratory procedure for confirmation and animal regain its normal value with the passage of time subjected to recovery. During followed up, two dogs were found to be dead because of CDV infection mixed with secondary bacterial infection in which one exhibited the nervous sign like teeth grinding, ataxia, convulsions and in coordination in body movements.
Only ten (22.22%) samples were found positive by RT-PCR using plasma, nasal and conjunctival swabs. CDV RNA was detected in 60% of plasma samples, 70% of nasal and 100% of conjunctival swab sample from lymphopenic dogs whereas the percentage was 13.33, 15,55, and 22.22 from a total of 45 samples. No amplicon of expected length was obtained from normal healthy dogs. On comparison of different fluid samples, the sensitivity of conjunctival swab was found to be highly significant followed by nasal swab and plasma.
In conclusion, Lymphopenia is the suggestive of clinical infection of dogs with canine distemper virus ad can help in presumptive diagnosis. It is not necessary that all lymphopenic dogs are distemper posit it requires further laboratory confirmtion. In this context, RT-PCR is test of choice with samples including conjunctival swabs and plasma.
Availability: Items available for loan: UVAS Library [Call number: 1034,T] (1).
5.
Electrophoretic Profile Of Cellular Proteins Of Staphylococcus Aureus From Mastitic Cattle
by Muhammad Zubair Munir | Prof.Dr.Irshad Hussain | Prof.Dr.Masood Rabbani | Prof.Dr.Zafar | Faculty of Veterinary Sciences.
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Publisher: 2008Dissertation note: Decline in milk production has. mostly been attributed to infectious agents that generally result in swelling of udder, changes in the constitutions of milk and finally induration of the udder which causing huge economic losses in livestock production. Mastitis is most important diseases of all the lactating animals. The most common Staphylococcus aureus which causes mastitis in cattle. Given the range of ill-effects of Staphylococcus aureus causing mammary infection in cows and buffaloes in Pakistan there is strong need for detection of strain involved. Likewise identification of clones with extensive geographic distribution wall provide insight into strain virulence and pathogenesis and also requiring public health intervention such as vaccination and antimicrobial restriction aimed at reducing the spread of the pathogen. In the present project the relatedness by elcctrophoretic profile of Staphylococcus aureus isolates involved in mastitis at dairy farns were studied, to compare the isolates of Staphylococcus aureus from different dairy farms. The samples from clinically/ subclinically affected at farms were collected, for isolation of Staphylococcus aureus, these samples were streaked onto Staph- 110 agar media plates and Staphylococcus aureus were be confirmed through biochemical and sugar fermentation tests. These conformed Staphylococcus aureus
Isolates were subjected to sodium dodecyl sulphate polycrylaniide gel electrophoresis. (SDS-PAGE) for electrophorctic protein profiling. The banding pattern of various isolates as against known molecular weight markers were recorded for interpretation of results.
It was concluded that protein profile is one of several methods for determining the relatedness or unrelatedness of bacterial strains. As immunization has been often attempted in efforts to control the mastitis, the diversity of Staphylococcus aureus strains and difficulties in vaccine development instruct that use of one or likely several strains in a herd or even in herds located in the same geographical region are recommended. Decision on representative strains can be made on basis of whole cell proteins, since such proteins of Staphylococcus aureus strain has already been shown, the isolates could be characterized by their total protein profiles and use of one or likely several strains of Staphylococcus aureus that are antigenically representative of the majority of the causative strains will overcome vaccine development difficulties.
Availability: Items available for loan: UVAS Library [Call number: 1047,T] (1).
6.
Detection Of Mycobacterium Tuberculosis And Mycobacterium Bovis From Spum And Blood Samples Of Human Using a Duplex PCR
by Asma Nawaz | Prof.Dr.Zafar Iqbal Ch | Dr.Azhar | Dr.Muhammad Younas Rana | Faculty of Veterinary Sciences.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2009Dissertation note: Tuberculosis is common infectious disease in the world. Mycobacterium tuberculosis is the most common cause of tuberculosis in the humans. Tuberculosis is endemic in Pakistan with about 1.5 million people infected. M.bovis is the major cause of gastrointestinal tuberculosis in humans.
The study was conducted in Lahore to compare 100 blood and 100 sputum samples from patients of active tuberculosis. The methods employed were conventional methods including Ziehl-Neelsen staining, culture on Lowenstein Jenson medium and biochemical tests. The Duplex PCR and conventional methods for diagnosis of tuberculosis caused by M.bovis and M.tuberculosis were compared on the sputum and blood samples. For M.tuberculosis and M.bovis the pncA gene and the species-specific 500-bp fragments were targeted in the Duplex PCR, respectively.
The sensitivity and specificity of Duplex PCR was found statistically significant in comparison to the conventional methods including Ziehl-Neelsen staining and culture for the differential diagnosis of tuberculosis caused by M.tuberculosis and M.bovis. Therefore Duplex PCR is a better choice of diagnostic test in the clinical setups where clinical urgencies necessitate a reliable, sensitive and specific test with the results in a short time period.
Availability: Items available for loan: UVAS Library [Call number: 1054,T] (1).
7.
Pathology Of Naturally Infected Broilers With Mycoplasma Gallisepticum And Its Diagnosis Through Pcr
by Aamir Islam | Dr.Asim Aslam | Prof.Dr.Mansur- | Prof.Dr.Zafar Iqbal Ch | Faculty of Veterinary Sciences.
Material type: ; Format:
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Publisher: 2009Dissertation note: Two hundred broiler birds (200) showing the clinical signs of respiratory signs from fifty (50) poultry farms located in and around Lahore District were analyzed for the detection of Mycoplasma gallisepticum. The tissue samples (trachea and lungs) were subjected to PCR using for Mycoplasma gallisepticum 16S rRNA gene amplification with a set of primers (MG14-F and MG13-R). Out of 200 samples, 86 were found positive with MG. These positive samples were further analyzed for histopathological changes. Lungs showed hemorrhages, congestion and massive necrosis. Lymphocytic infiltration and oedema was also observed in lungs sections. Liver showed coagulative necrosis around the central vein, congestion and infiltration of lymphocytes. Similarly, heart section revealed necrosis and degeneration in cardiac muscles. Trachea revealed the epithelial and mucosal infiltration with lymphocytes. Hypertrophy of epithelial mucosa and catarrhal exudates recorded in trachea. Sloughing of the mucosa and sub mucosa of varying degree was noted in trachea. Few birds showed no obvious changes in the organs but were positive on PCR analysis.
Availability: Items available for loan: UVAS Library [Call number: 1055,T] (1).
8.
Effect Of Total Particulate Matter From Commercial Cigrettes On Wound Healing In Mice
by Sidra Abdil Rahman | Prof.Dr.Muhammad Ashraf | Dr.Sheryar Afzal | Prof.Dr.Zafar Iqbal Ch.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2009Dissertation note: ACigarette smoking is considered as the most important cause of death in developed countries. Wound healing in the skin is a multifarious orchestration of cellular processes and cigarette smoking may grounds for delayed wound healing. The aim of this study was to investigate, the plausible association between exposures of cigarette total particulate matter (TPM) on wound healing. The invivo mice model of wound healing was customized for determinenation of assorted events of wound healing. A total of 49 adult mice separated in seven groups and kept under standard conditions of ventilation and temperature. Four brands of commercially available cigarette were selected for the current study. All the treated animals were exposed to TPM ointment for 12 days. A highly considerable diminution in wound closure was pragmatic among all TPM treated groups from day 6 to day 8 post-wounding. Histological investigations unveiled a noteworthy impede in the outcome of re-epithelialization, dermal matrix regeneration and maturation of collagen bundles among all TPM-exposed wounds. Delayed commencement of neovascularization was pragmatic among TPM treated groups, at day 12 post wounding. Quantification was done with the help of scan probing image processing software and image processing system. Abbot curve, angular spectrum and different other parameters of 3D surface behavior of wounds revealed a very highly significant reduction in angiogenesis at day 6 and day 8 post-wounding, which points that application of TPM instigate extensive delay in trigging the progression of angiogenesis, resulting delayed onset of wound healing. These annotations validate the damaging effects of TPM on the healing of wounds and excessive use of TPM may lead to the production of chronic wounds and oral ulcers.
Availability: Items available for loan: UVAS Library [Call number: 1071,T] (1).
9.
Prevalance And Anthelmentic Activity Of Indigenous Plants Against Trichostrongylus Of Sheep In District Zhob
by Nasib Ullah | Dr.Muhammad Lateef | Prof.Dr. Azhar Maqbool | Prof.Dr.Zafar | Faculty of Veterinary Sciences.
Material type: ; Format:
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Publisher: 2010Dissertation note: A Trichostrongylid is gastrointestinal nematode which causes the heavy economic losses to the livestock particularly sheep. A total 240 gastrointestinal tracts of sheep from district Zhob abattoir were collected. These samples were isolated and identified for trichostrongylid nematodes. The overall prevalence was 50%(120/240), of which 39.1% in male and 60.8% in female sheep were recorded at slaughter-house. Plants were collected from area of district Zhob. These plants were Identified and authenticated by botanist .The crude aqueous methanolic extract of the plants were used for in-vivo studies. Eighty sheep of either sex, aged between three to six months and naturally infested with Trichostrongylid nematodes (including trichostrongylius spp. I-Iaemonchus contortus, cooperia etc) were selected and managed separately for the experiment. These sheep were divided into 4 groups A ,B,C and D. Group A was contained 10 sheep and was untreated and considered as control. Thirty (30) were kept in Group B, were further sub divided into three equal groups i.e. BI, B2 and B3 and treated with different levels of Chenopodium album @ 1, 2 and 3 g/kg body weight respectively. Group C having thirty (30) was also sub divided into three equal groups i.e. Cl, C2 and C3 and treated with different levels of Artemisia brevifolia @ 1, 2 and 3g/kg body weight respectively. Group D was treated with Levamisole @ 7.5 mg/kg body weight. Faecal egg count reduction was criterion for evaluation. Faecal samples were collected before treatment on day 0 and on day 3, 5, 7, 10 and 14 post treatments for EPG count.
The results of EPG for animals in group B1 at day 0 was 1325. This rate was reduced to 1250, 1125, 995 and 702 at day 3, 7, 10 and 14, respectively. Similarly EPG of sheep in B2 were 1280 at day 0 and reduced to 1205, 1202, 1001 and finally to 690 at day 3, 7, 10 and 14 respectively. Best results in B group against the nematodes were in B3 in which Chenopodium album was used 3g/kg bw. The results were 1250, 1231, 1145, 590 EPG at day 0, 3, 7, 10 and 14. The sheep in group Cl showed 1203 EPG at day 0, when treated with Ig/kg bwArtemisia brevifolia, the EPG was reduced to 1173, 1115, 700 and 528 on day 3, 7, 10 and 14 respectively. Second level of treatment C2 of
Artemisia brevifolia which was given @ 2g/kg bw initially contained reduced to 1202 EPG, on day 3, 7, 10 and 14 the EPG counts were 1020, 631, 546 and 459, respectively.
Highest dose of Artemisia brevfolia was 3g/kg body weight to sheep in group C3. On day zero the EPG count was 1196. On day 3, 7, 10 and 14th day the EPG count decreased to 1079, 905, 528 and 396 respectively. The sheep in group D, treated with recommended dose of Levamisole showed 1138 EPG prior to medication, which reduced remarkably to 681, 536, 357 and 147 on day 3, 7, 10 and 14 respectively. Although no untoward effects of plants were observed but best EPG reduction results (87.08 %) were observed in Levamisole as compared to treatment of 3gm/kg b.w Chenopodium album (51 .03 %) and Artemisia hrevfolia 3g/kgbw (66.88 %).
CONCLUSIONS
A wide variety of plants are naturally available in the Indo-Pakistan subcontinent which possess narrow or broad spectrum anthelmintic activities. No doubt this is true in other regions of the world as well where gastrointestinal parasitism is an important problem in livestock keeping, and the availability of commercial drugs may be limited. Conventionally, trichostrongylids has been tackled with use of synthetic anthelmintic but owning to development of anthelmintic resistance against major groups of anthelmintics viz., benzimidazole, Levamisole and avermectins, people are looking for alternatives to synthetic chemicals.
The phytochemical analysis of these plants and controlled anthelmintic trials along with contemporary knowledge of parasite control strategies may offer new opportunities for effective and economical control of parasitic diseases. So these plants can be better alternative for synthetic chemicals.
Quality control extracts of Artemisia brevifolia and Chenopodium album or possibly isolated bioactive compounds could be a promising alternative to conventional anthelmintics fbr the treatment of gastrointestinal trichostrongylids of small ruminants in the future. Such a treatment could be used in control strategies against gastro intestinal nernatodes in organic and conventional production systems. Further research is needed for studies on the bio active constituents, as well as on the reproducibility, dosage, application regime. toxicity and effectiveness of Artemisia brevifolia and C'henopodium a/bums in other host species and against other economically important gastro intestinal nematodes species.
RECOMMENDATIONS
It is recommended that further research could be carried out on large sample size in different seasons of the year and large number of plants, identification of active principles of plants with proven anthelmintic activity, standardization of dose and toxicity studies for drug development. In addition to this, large number of samples of the same plant from different geographic areas should be subjected to experimentation keeping in view the possibility of differences in chemical composition of soils.
Availability: Items available for loan: UVAS Library [Call number: 1075,T] (1).
10.
Comparison Of Different Diagnostic Techniques For John'S Disease In Small Ruminants
by Saba Badar | Prof.Dr.Zafar Iqbal Ch | Dr. Mansur-ud-Din | Dr.Asim Aslam | Faculty of Veterinary Sciences.
Material type: ; Format:
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Publisher: 2010Dissertation note: Paratuberculosis is one of the most hazardous infectious diseases, causing heavy economic losses due to poor health, low productivity and high fatality rate among domestic and wild ruminants. Mycobacterium avium subsp. paratuberculosis is the etiological agent of Bovine Johne's disease. In this study PCR were used to detect the presence of the Acid Fast Bacillus Mycobacterium paratuberculosis, in the intestinal tissues and Mesenteric Lymph Nodes of small ruminants causing Paratuberculosis. PCR was compared to HEY medium culture on the Herrold's Egg Yolk Media.
The samples were collected from Lahore Slaughter house and brought to the Molecular Pathology Laboratory at the Department of Pathology, University of Veterinary and Animal Sciences, Lahore. The study was conducted to compare PCR and the HEY medium culture for the diagnosis of paratuberculosis caused by M avium subsp. Paratuberculosis. A total of 500 tissue samples, 250 of the ileum and 250 of the mesenteric lymph nodes were collected randomly for the identification of Johne's disease. All samples were inoculated on the HEY medium prepared in the same laboratory aseptically. Followed by DNA extraction through the Kit method then run the PCR for insertion sequence IS 900, specific 626 bp fragment, were targetted in the genome of M paratuberculosis.
The results of the study showed more samples detected positive by PCR as compared to conventional culture methodology. Also they showed in the mass of 500 tissue samples that more bacilli are prone to the samples of small intestines than associated mesenteric lymph nodes. Regarding the sensitivity of the two techniques the PCR seemed more sensitive to detect the mycobacterium in the tissues than the conventional, laborious and time consuming HEY medium culture technique; though culture has been used as golden standard in this study also. When statistically analyzed results were insignificant due to small sample size.
The study will help in comparison of the two latest techniques for the diagnosis of M paratuberculosis, to check the validity of the better technique. In this study the sensitivity and specificity of PCR was checked and compared with culture on the HEY medium staining for the diagnosis of paratuberculosis in small ruminants.
Availability: Items available for loan: UVAS Library [Call number: 1077,T] (1).
11.
Immunohistochemical And Pathomorphological Studies Of Chronic Granulomatous Enteritis (John'S Disease) in Bovines
by Muhammad Shahid | Prof.Dr.Zafar Iqbal Ch | Dr.Asim Aslam | Prof.Dr.Khushi Muhammad | Faculty of Veterinary Sciences.
Material type: ; Format:
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Publisher: 2010Dissertation note: Paratuberculosis, a disease caused by Mycobacterium paratuberculosis is a peril for both livestock and human beings. The present project was designed to study the pathmorphological changes induced by the organism and standardize more reliable diagnostic techniques to identify the M paratuberculosis. Tissue samples from ileurn and mesenteric lymph nodes were randomly collected from 1 50 cattle and buffalo, each in present study that was conducted in Lahore.
Gross lesions were recorded on a Performa. The samples were subjected to acid fast staining of smears from pellets after density gradient centrifugation and paraffin embedded tissue sections. All the samples also subjected to polymerase chain reaction and immunohistochemistry. The smears prepared from bacterial pellets of mucosal and cortical scraping of terminal ileum and MLN were stained indicated 11.4 % small intestine and 12.7% lymph nodes of cattle's and 8.7% and 10.7% lymph nodes of buffalo's tissue samples were positive. ZN staining of paraffin embedded tissue showed 8.0 % small intestine and 10% MLN of cattle's and 6.0 % of small intestine and 8.7% MLN in buffalo's tissue samples were positive. On basis of PCR 5.4% intestinal tissue samples and 6.0% MLN of cattle were positive. 3.4% intestinal tissue samples and 07(4.7%) MLN of buffaloes were positive. In buffaloes 4.0% intestinal tissue samples and 6.0% MLN were positive by IHC. In cattle 6.7% intestinal tissue samples and 8.0% MLN tissue samples were positive by IHC. In cattle, 27/150(18.0%) animals showed lesions in both intestine and mesenteric lymph nodes while 5/32 (15.7%) animals showed lesions in lymph nodes only. Out of 27/150(18.0%) intestinal tissue samples, 20/27 (74.1%) samples showed corrugation of the intestinal mucosa while 7/27 (26%) showed diffuse thickness.
In buffalo, 24/150 (16.0%) animals showed lesion in both intestine and mesenteric lymph nodes while 2/26 (7.7%) animals showed lesion in lymph nodes only. Out of 24 intestinal tissue samples, 19/24(79.2%) with gross lesion, samples showed corrugation of the intestinal mucosa while 5/24(20.9%) showed diffuse thickness.
In histopathology 20/27 samples of cattle showed focal granulomatous lesions while 7/27(26%) samples showed sever infiltration of macrophages and lymphocytes while 28/32(87.5%) lymph nodes showed infiltration of paracortical and cortical region of the lymph nodes with macrophages ,lymphocytes and multinucleated giant cells While 4/32 (12.5%) samples showed moderate infiltration of macrophages.
In buffaloes 19/24 (12.7%) samples showed focal granulomatous lesions while 5/24 (20.9%) samples showed sever infiltration of macrophages and lymphocytes while 22/26 (84.7%) lymph nodes showed infiltration of paracortical and cortical region of the lymph nodes with macrophages ,lymphocytes and multinucleated giant cells While 4/26 (15.4%) samples showed moderate infiltration of macrophages.
The sensitivity and specificity of immunohistochemical method was found significant in comparison Ziehl-Neelsen staining and histopathology for the diagnosis of paratuberculosis in cattle and buffaloes.
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12.
Polymerase Chain Reaction And Restriction Fragment Length Polymorphism (Rflp) By Using Ssu-r DNA Amplification for the Species Specific Diagnosis of Trypanosomiasis in Horses
by Naveed Sabir | Prof.Dr.Zafar Iqbal Ch | Dr.Asim Aslam | Prof.Dr.Khushi Muhammad | Faculty of Veterinary Sciences.
Material type: ; Format:
print
Publisher: 2010Dissertation note: In the current research project, a pari-trypanosome polymerase chain reaction (PCR) was optimized by using 18S single sub unit ribosomal DNA amplification and restriction fragment length polymorphism (RFLP) was also optimized and evaluated for the species specific diagnosis of the trypanosomiasis in horses.
Blood samples from one hundred (100) suspected horses were collected aseptically from different localities of Lahore. Fresh blood smear was prepared from each sample. After drying and fixing with absolute methanol, the slides were stained with Giemsa stain. Microscopic examination of stained blood smears revealed 8 positive samples out of one hundred (100) suspected horses. Polymerase chain reaction (PCR) was carried out on the same trypanosomiasis suspected blood samples to evaluate its sensitivity. Genomic DNA was extracted by using Genomic DNA Purification Kit (Fermentas mci., USA). The PCR was performed in a 50 tl reaction mixture. The tubes containing PCR mix were subjected to amplification cycles in a thermocycier after adjusting the amplification conditions.
After completion of the amplification cycles, the PCR product was characterized by 1.2 % agarose gel electrophoresis along with 100 bp DNA ladder to estimate the size of the PCR product and the gel was photographed with a Polaroid camera. PCR gave a higher percentage of positive cases i.e. 21% as compared to microscopic examination. Semi-nested polymerase chain reaction was carried out on product of the first run amplification by using same reaction mixture and amplification conditions except for template DNA. In case of semi-nested PCR 1 tl of the simple PCR product was used. Semi-nested PCR gave 100% (21/21) results. Restriction fragment length polymorphism (RFLP) analysis was conducted on nested products of the positive samples. A reaction mixture of 20 1iJ was used and samples were incubated over night at 37 °C in an incubator. The restricted products were characterized by 2 % agarose gel electrophoresis along with 100 bp DNA ladder and photographed with Polaroid camera. Restriction fragment length polymorphism (RFLP) analysis of the nested products revealed that none of the species including T. congolense, T. theileri, T. brucei and T. vivax was found in all (2 1%) positive animals having trypanosoma infestation.
It can be concluded from current study that a pan-trypanosome polymerase chain reaction is a superior and sensitive test as compared to Giemsa stained blood smear examination. The test can not only be used for early diagnosis of the trypanosomiasis but it can also be used to screen out the carrier animals those act as a reservoir of the infection for the horses and other susceptible animals. The advantage of this test is its sensitivity, universal applicability and the existence various possibilities for restriction enzyme analysis of the amplified region depending on the trypanosome species.
Availability: Items available for loan: UVAS Library [Call number: 1079,T] (1).
13.
Effect Of Two-Stroke Auto Rickshaw Smoke Solution On Wound Healing In Mice
by Aqeela Iqbal | Prof.Dr.Muhammad Ashraf | Dr.Muhammad Ovais Omer | Prof.Dr.Zafar.
Material type: ; Format:
print
Publisher: 2009Dissertation note: The use of 3-wheel auto-rickshaws powered by a 2-stroke engine is widespread in south Asia; exhaust from these vehicles may cause different types of toxicities resulting in different pathologies. The aim of this study was to explore the association between exposure to 2-stroke auto-rickshaw smoke solution (2SARSS) and wound healing. The in vivo model of wound healing was customized to evaluate different stages of wound healing. A total of 56 mice were used of aged 4-5 weeks. After anesthetizing and hair removing from dorsum side of mice a wound of 3mm diameter was generated with the help of a sterile wound puncture. Smoke was collected in the lOOml distilled water from six 2-stroke rickshaws and then these solutions were filtered by syringe filters. 10% ointment of all the samples was prepared in Petroleum Jelly and applied to the experimental group, petroleum jelly to positive control group, no treatment was applied to negative control. Macroscopic evaluation was done by using high resolution camera photographs and measurement of wound by vernier caliper on alternate days. At the end of the study (day 12) histological evaluation was done when the control group wounds were healed. Histological examination revealed a significant delay in the outcome of re-epithelialization, dermal matrix regeneration, and maturation of collagen bundles among all 2SARSS-exposed wounds. Delayed activation of neovascularization was seen in the 2SARSS-treated groups at day 12 post-wounding. The Abbot curve, angular spectrum, and several other 3D surface parameters of reverse wound topographies revealed a highly significant reduction (p<O.OOl) in angiogenesis. These results demonstrate that application of 2SARSS causes a substantial delay in the progression of angiogenesis, resulting in delayed onset of wound healing. These observations validate the damaging effects of 2SARSS on wound healing. Thus, people who are directly or indirectly exposed to this toxic exhaust are expected to have delayed wound healing, which could result in chronic wounds.
Availability: Items available for loan: UVAS Library [Call number: 1083,T] (1).
14.
Tissue Residue Studies Of Enrofloxacin In Broilers Chicks
by Irfan Irshad | Prof.Dr.Zafar Iqbal Ch | Dr.Asim Aslam | Prof.Dr.Muhammad Athar Khan | Faculty of Veterinary Sciences.
Material type: ; Format:
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Publisher: 2009Dissertation note: Poultry industry is the second largest industry of Pakistan. Antibiotics are enormously used in poultry both for prophylactic and treatment purpose. Irrational use of antibiotics in poultry industry has led to the serious concern among the general public. It has also resulted in emergence of drug resistance in many susceptible organisms. The present study has therefore been planned for quantitative detection of Enrofloxacin residues; in tissues (liver, kidney, fatty tissues and muscles) of broiler birds. So, the present study was designed to detect Enrofloxacin residues in tissues of birds reared under experimental conditions and routinely slaughtered at different poultry shops. The study was completed in two phases. In phase-I, tissue samples from 75 broiler birds reared at Department of Pathology, University of Veterinary and Animal Sciences, Lahore were analyzed for quantitative detection of Enrofloxacin by HPLC. In phase-TI, the 25 broiler birds were purchased from various poultry shops of different local markets of Lahore. Tissue sample (Liver, Kidney and thigh muscles) from these broiler birds were also analyzed for quantitative detection of Enrofloxacin by HPLC. In experimentally reared birds, the highest concentration of Enrofloxacin observed was 306ng/g. In the birds injected with Enrofloxacin intramuscularly the overall highest concentration was 68ng/g. The concentration in kidney, liver and thigh muscles was in the range of 28-64 ng/g, 26-63 ng/g, 26-68ng/g in kidney, liver and thigh muscles respectively in birds injected with drug intramuscularly. The drug residues were detected up to 120 hours post treatment in intramuscularly injected birds. In orally treated birds level of Enrofloxacin in the kidney, liver and muscles were between 56-2 17 ng/g, 29- 306 ng/g ,27- 170 ng/g. The residues were detected up to 96 hours post treatment in birds given Enrofloxacin orally. The result of phase-Il showed that among the 75 market samples, 10 (40%) muscles, 8 (32%) liver and 7 (28%) kidney samples showed the Enrofloxacin residues. Out of 25 samples in which Enrofloxacin residues were detected 20 (80%) samples showed the residues concentration above MRL.
This study helped us in drawing true picture about Enrofloxacin drug residues in poultry meat and it is clearly indicated that proper withdrawal time is not being observed while marketing birds. This poses a great health concern for end consumer.
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15.
Study Of Pathogenesis Of Mycoplasma Gallisepticum In White Leg Horn Layer
by Mubasher Rauf | Prof.Dr.Zafar Iqbal Ch | Dr Aftab | Dr.M.Younus Rana.
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; Literary form:
not fiction
Publisher: 2010Dissertation note: In first part of present study 380 samples were collected from clinically suspected cases of layers suffering from respiratory diseases in and around Lahore. Samples were subjected for mycoplasma isolation by using Frey's medium. Plates with positive growth revealed characteristic colonies on 8th day post inoculation that reached maximum in size and growth at 15th day post inoculation. Out of 380 samples, 104 (27.36%) samples were positive on culture. Isolates were identified through growth inhibition test (GIT) by using hyper immune sera raised in rabbits. Isolates were further confirmed by PCR. Similarly, tracheal swabs and tissue samples of lungs and trachea collected under refrigeration were also subjected for DNA analysis. Out of 380 samples 264 (69.5%) were positive on PCR analysis. By comparing two diagnostic techniques it was found that PCR was more sensitive and reliable technique for screening of Mycoplasma gallisepticum.
In second part of study experiment isolates were analyzed for protein profile of Mycoplasma by standardization of two techniques Sodium dodecyl sulphate polyacrylamide gel (SDS-PAGE) and western blotting. These techniques help to find out any antigenic variation in prevailing strain of mycoplasma. During our study five bands of protein were detected with molecular size of 32.35 kDa, 43.65 kDa, 52.48 kDa, 64.56 kDa and 70.8 kDa. These proteins were extracted from whole cell of Mycoplasma gallisepticum isolates. On comparing the molecular sizes it was found that isolated species showed low antigenic variation, analyzed by SDS-PAGE. Western blot was used to determine the specific protein of Mycoplasma gallisepticum with the use of specific polyclonal antibody raised in rabbit. The positive reaction site was shown on nitrocellulose membrane confirming target species of CRD.
During third part of present study, it was concluded that aerosol route of infection causes early disease, followed by intra tracheal and per-oral route respectively. The severity of infection was found more in aerosol and intra tracheal routes of inoculation than per-oral route which was found to be very mild. The general gross lesions observed in the above two groups were hemorrhages in trachea with mucous plug. There was air sacculitis, hemorrhages in the lungs, salpingitis and putrefied eggs in the ovary. On histopathological examination lesions were found in trachea, lungs and oviduct.
Re-isolation was carried out to confirm antigen in experimentally inoculated birds. Paraffin embedded sections of trachea, lungs and oviduct were processed for immunohistochemical examination in order to confirm the antigen of Mycoplasma gallisepticum within tissue. A positive immunochemical reaction was found in lungs and oviduct. Which represents that antigen was same as inoculated during study of pathogenesis.
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16.
Epidemiology And Controls Of Coccidiosis In Cattle
by Razia Sultana | Prof. Dr. Azhar Maqbool | Prof.Dr.MAnso | Prof.Dr.Zafar Iqbal Ch.
Material type: ; Format:
print
Publisher: 2009Dissertation note: Field study was conducted from September, 2007 to August, 2008 and a total of 2700 rectal faecal samples were collected from cattle farms of 3 categories i.e. Government Dairy Farm, Military Dairy Farm and Peri Urban Dairy Farms (Gawala Colonies) Lahore. Seventy five random samples were collected from each category of farms on monthly basis. The results of field study showed that overall prevalence of coccidiosis in cattle was 54.55%. Prevalence of coccidiosis in cattle at Military Dairy Farm Lahore was the highest (65.33%) during Autumn followed by summer (52.66%) then winter (47.66%) whereas the lowest (34.00%) during spring season. The highest (56.66%) prevalence was observed in animals between 6 & 12 month, whereas the lowest (46.33%) in animals under 6 months age. Prevalence of coccidiosis above 1 year of age was 50.66%. No coccidial oocysts was detected in calves less than 15 days old. In female animals prevalence was 51.22%.In the present study, the maximum oocyst per gram of feces (OPG) count was 65,000 whereas the minimum count was as 2000. The count was variable in different age groups and found to be decreasing in adult animals. The mean OPG in group A (under 6 month), group B (6 month to one year) and C (above one year) was 44000, 38000, and 22000, respectively. The four species of Eimeria were identified in all age groups i.e. E.bovis (29.28%) E.zuernii (26.03%) E. cylindrica (23.42%), E. ellipsoidalis (21.25%).
The results of field study showed that prevalence of coccidiosis at Government Dairy Farm, Lahore was the highest during autumn (49.33%), followed by summer (44.33%), then winter (38.33%) where as the lowest during spring (30.33%). The highest (62.66%) month wise prevalence of coccidiosis was noted during August whereas the lowest (28.00%) during April. The highest ( 45.33%) prevalence of coccidiosis was observed in animals aged between 6 to 12 months, followed by 41.35% in animals under 6 months of age whereas the lowest (36.00%) above I year. Female animals were more frequently affected (41.28%) than males (39.50%).In the present study, the maximum OPG count observed was 55,000 and the minimum count as 2500. The counts were variable in different age groups and found to be decreasing in adult animals. The mean OPG of group A, B, C was 42,000, 35,000 and 20,000 . In the present study five species of Eimeria were E.bovis. E. zuernii E. cylindrica, E. subspherica, E. ellipsoidalis.
The results of field study showed that prevalence of coccidiosis at Peri Urban Dairy Farms (Gawala colonies), Lahore was 71.55%. Month wise prevalence was the highest during August (90.66%) whereas the lowest (48%) during April. The seasonal prevalence indicated that it was the highest during autumn (84.00%), followed by summer (78.33 %), then winter (69.33%) whereas the lowest during spring (50.00%). The highest prevalence of coccidiosis (80.66%) was observed in animals under 6 months of age, whereas the lowest (62.33%) in animals above I year. Prevalence of coccidiosis in animals aged between 6months to 1 year was 71.66%. No coccidial oocysts were detected in calves less than 25 days old. Prevalence of coccidiosis was higher (74.61%) in females than in males (63.60%). In this study, the maximum OPG count observed was 65,000 and the minimum count as 2800. The counts were variable in different age groups and found to be decreasing in adult animals. The mean OPG of group A, B,C was 48,000, 38,000 and 23,000 respectively.
Age wise analysis of Eimeria species showed that above mentioned five species were found in all age groups and most predominant species was E.bovis (26.39%) followed by E. zuernii (19.87%), E. cylindrica (23.60%), E.ellipsoidalis (18.63%), whereas the lowest prevalence of E.subspherica (11.49%)was noted (Table 16). The counts were variable in different age groups and found to be decreasing in adult animals. There was inverse correlation of OPG and the age of animals.
The overall prevalence of coccidiosis was the highest during autumn (66.22%) followed by summer (59.66 %) then winter (51.77%) whereas the lowest in spring (38.22). The role of Meteorological data i.e. temperature, humidity and rain fall on the prevalence of disease was also studied. The bionomical showed that humidity and rain fall played a very important role in the causation and spread of disease and also help in the development of sporulated oocyst. Increased temperature showed higher prevalence of disease.
The results of histopatholgical studies showed that there was an increase cellular infiltration of leukocytes, cellular debris in most of intestinal portion.
Results of therapeutic trials by using toltrazuril, amprolium, sulphaquinoxaline, lasalocid are presented in table 17. The result of therapeutic trials showed that efficacy of toltrazuril was better than amprolium, sulphaquinoxaline and lasalocid. No clinical signs of disease were observed in treated animals while in diseased animals signs of disease were observed i.e. animals showed diarrhoea, loss of weight gain. From the results it was noted that efficacy of toltrazuril was better than other drugs . Statistically, there was no significant difference between efficacies of all four drugs. The efficacy of per oxygen based disinfectant was higher as compare to oocide while non- treated animals showed clinical signs of disease. Statistically, there was no significant difference between efficacies of both disinfectants
Result of chemo prophylactic products are presented in table 19. It was noted that sonicated vaccine showed high antibody titer as compare to non- sonicated vaccine.
Result of the challenge experiments revealed that the inactivated sonicated vaccines gave 100% protection to the challenge calves. Their faeces were normal and no clinical sign was recorded even 42 days post vaccination. Few remaining live oocysts were not able to produce the disease in calves.
The weight gain of treated animals was higher as compare to non-treated animals. The FCR value in treated animals was better than non treated animals
" Prevalence of coccidiosis was the highest during autumn followed by summer where as the lowest during spring. Farm wise prevalence of coccidiosis indicated that it was higher in Peri Urban Dairy Farms followed by Military Dairy farm where as the lowest at Government Dairy farm.
" Prevalence of coccidiosis was higher in calves below 9 months of age than above 9 months. All the animals examined for coccidian were naturally infected with coccidiosis. These animals were not experimental calves and prevelance of infection was based on random selection of animals. Overall Prevalence of coccidiosis was slightly higher in females than male. Species wise prevalence indicated that Eimeria bovis is more pathogenic than other species.
" Results of chemotherapeutic trials showed that among the four drugs used i.e. Toltrazuril, Amprolium, Sulfaquinoxaline and Lasalocid. Toltrazuril showed the highest efficacy followed by Amprolium, where as Lasalocid showed the lowest efficacy. No side effects of these drugs were noted when were given at their recommended dose rate and marked clinical improvement in animals was noted after treatment.
" Two disinfectants were tried. Per oxygen based disinfectant showed better results than. Oocide disinfectant.
" Histopathological studies showed inflammatory granulocytic infiltration of the mucosa and cellular debris in most of intestinal portions. There were necrosis of villi and degeneration of villi. Haemorrhages in mucosa and sub-mucosa were noted. Some of the glands in the sub-mucosa of intestine showed degeneration & necrosis.
" Indirect Haemagglutination (IHA) antibody titer was higher in calves vaccinated with inactivated sonicated vaccines as compared to the calves vaccinated with inactivated sporulated vaccine. Results of the challenge experiments revealed that the inactivated sonicated vaccines gave protection to the challenge calves. Their faeces were normal and no clinical sign of disease were observed even 42 days post vaccination.
" Weight in infected group was reduced. After treatment, high weight gain was reported in treated animals than control group.
Recommendations:
" Overcrowding should be avoided.
" Provide good hygienic and managemental conditions in farms.
" Proper drainage of rain.
" Feeders and wateres should be above the level of the ground.
" Regular use of coccidiostats is the need of the day.
" Diseased animals particularly with diarrhoea should be separated from healthy animals.
" Stocking density should be according to recommended of world Association of Parsitologists.
" Contaminated faeces should be properly disposed off.
" Grazing of animals during rainy season should be avoided.
" Animals should be provided well balanced nutritive food.
" Entry of visitors in the livestock farms should be restricted
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