Molecular Diagnosis And Therapeutic Trials On Fasciolosis In Sheep And Goats
Material type: Book ; Format:
Publisher: 2012 Dissertation note: The fasciolosis in sheep and goat is an important and infectious disease of ruminants characterized by dullness, weakness, lack of appetite, pallor and edema of mucosa and death of animal. The accurate diagnosis of Fasciolosis remained a difficult task for the field practitioners and the right choice of medicine.
For this purpose 600 sheep and goats was examined for fasciolosis. The fecal samples were collected directly from rectum while bile samples were collected from various private and public slaughter houses of districts Okara, Sahiwal and Lahore. Fasciola egg was identified by standard text while molecular conformation of Fasciola spp. was done by using PCR. For therapeutic trials, sixty animals (n=30 sheep; n=30 goats) positive for Fasciolosis was equally divided into A, B and C groups and each group comprising of 20 animals (n=10 sheep; n=10 goats). The animals of group A was given Zanil (Oxyclozanide) orally @ 1ml per 3kg body weight; the animals of group B was given Albendazole orally @ 1ml per 3kg body weight and the animals of group C was given the grounded powder of Neem leaves (Azadirachta indica) @ 5-6 teaspoon (25-30gm) orally. Efficacy of drugs was measured on the basis of disappearance of Fasciola eggs at 3, 7 and 14 day of post-medication. Data on the prevalence of fasciolosis was analyzed by Pearson's chi-square test while therapeutic trials data was analyzed with one way ANOVA using Statistical Package for Social Sciences (SPSS); P < 0.05 was considered significant.
To study fasciolosis in sheep and goats, samples were collected from Lahore, Okara and Sahiwal. The collected samples were processed at the Medicine laboratory at UVAS, Lahore and LPRI laboratories of Bahadurnagar, Okara. In this study first of all the fecal and bile samples were examined through microscope then these samples were processed by PCR test. No study was conducted previously on the molecular diagnosis of fasciolosis in sheep and goats in Pakistan, this is the first report for the molecular diagnosis of fasciolosis. In all F. hepatica positive samples approximate 300-bp fragment was generated on gel tested with DSJF/DSJ3 primers for sheep and goats. All positive samples by microscope were also positive by PCR, but out of the 92 samples positive (43 in sheep; 49 in goats) by PCR test, out of these 14 (7 in sheep; 7 in goats) were negative by microscopy. This study provides the base line data for the molecular diagnosis of fasciolosis in sheep and goats in Pakistan.
Overall prevalence of fasciolosis in sheep in both fecal and bile samples in various districts of Punjab was calculated. Out of 300 samples (150 fecal; 150 bile) of sheep, F. hepatica prevalence rate was 12% (36 samples) by microscopic examination while 14.33% (43 samples) by PCR test. On the basis of microscopic examinations districts wise prevalence rate was 11, 12 and 13% in Lahore, Okara and Sahiwal respectively and by PCR test prevalence rate was 13, 14 and 16% in Lahore, Okara and Sahiwal respectively. Prevalence of F. hepatica was more in Sahiwal than Okara and Lahore in sheep in both fecal and bile samples on the basis of microscopic examination and PCR.
Overall prevalence of fasciolosis was also calculated in goats on the basis of microscopic examination and PCR in both fecal and bile samples. Among 300 samples (150 fecal; 150 bile) of goats F. hepatica prevalence was 14% (42 samples) by microscopic examination while 16.33% (49 samples) by PCR. On the basis of microscopic examinations the prevalence was 13, 15 and 14% in Lahore, Okara and Sahiwal districts respectively while on basis of PCR test prevalence was 15, 18 and 16% in Lahore, Okara and Sahiwal districts respectively. Prevalence of F. hepatica was more in Okara than Sahiwal and Lahore in goats in both fecal and bile samples on the basis of microscopic examination and PCR.
The eggs per gram (EPG) values and efficacy of Oxyclozanide (Zanil), Albendazole and Neem leaves against fasciolosis in 30 positives sheep was recorded at 3, 7 and 14 days after the treatment. As group A was treated with oxyclozanide (Zanil), its efficacy was 50, 75 and 90% at 3rd, 7th and 14th day respectively after treatment. The efficacy of albendazole was 46, 65 and 81% at 3rd, 7th and 14th day respectively while the efficacy of Neem leaves was lowest that was 15, 30 and 42% at day 3rd, 7th and 14th day respectively. The efficacy of oxyclozanide and albendazole was significantly higher (p= 0.000) than Neem leaves. In sheep against the fasciolosis Oxyclozanide significant effect on EPG followed by albendazole and Neem leaves.
The EPG and efficacy of Oxyclozanide (Zanil), Albendazole and Neem leaves were determined in 30 goats against fasciolosis at 3, 7 and 14th day of post treatment. The efficacy of oxyclozanide was 57, 76 and 90% at 3, 7 and 14th day of post treatment, respectively. The efficacy of oxyclozanide was higher than Albendazole against the fasciolosis. The efficacy of albendazole was 55, 68 and 79% at day 3, 7 and 14th, respectively. Efficacy of Neem leaves against fasciolosis was lower as compared to other two drugs, it was 15, 23 and 40% at day 3, 7 and 14th, respectively. When the efficacy was compared at different days, Oxyclozanide reduced the EPG significantly, followed by Albendazole and Neem leaves.
From this study it was concluded that PCR is more reliable technique than microscopic examination for the diagnosis of fasciolosis in sheep and goats. Microscopic technique provides the base line data for the diagnosis of fasciolosis in sheep and goats but PCR provides help to conduct further molecular diagnosis on fasciolosis as well as others parasitic diseases. Secondly fasciolosis which is an important parasitic problem of ruminants is significantly prevalent in sheep and goats in Lahore, Okara and Sahiwal districts of Punjab. Oxyclozanide is the most effective drug while albendazole and neem leaves (Azadirachta indica) are relatively less effective against fasciolosis in sheep and goats.
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Prevalence And Therapeutic Studies On Diabetes Mellitus In Pet Dogs At Lahore.
Material type: Book ; Format:
Publisher: 2012 Dissertation note: Diabetes mellitus is one of the most important maladies of endocrine glands in dog and cats. It is due to the insufficient production of insulin by the pancreas. The multi-factors such as obesity, feeding, exposure to toxic drugs etc. are involved for the onset of this chronic disease. The basic energy fuel of the body is glucose. In the absence of insulin the glucose level in the blood is increases which can be noticed as hyperglycemia (high blood glucose level). There are the three types of diabetes mellitus which includes insulin-dependent diabetes mellitus (type I or IDDM), noninsulin-dependent diabetes mellitus (type II or NIDDM) and secondary diabetes mellitus. The dogs play a vital role in our society.
The present study was conducted on diabetes mellitus to study its prevalence, its effect on various blood parameters and to evaluate the efficacy of insulin therapy in pet dogs in Lahore, Pakistan. The 500 blood samples were collected from the dogs of age ?5 years, presented at Pet centre of the University of Veterinary and Animal Sciences (UVAS) Lahore and various private clinics in Lahore, and was processed in Medicine Laboratory of UVAS, Lahore. The blood glucose level was measured with the aid of glucometer. Out of 500, 16 (n=5 male & n=11 females) were positive for diabetes mellitus. The 3.2% prevalence of diabetes mellitus was recorded in pet dogs in Lahore. The study concluded that the prevalence of diabetes mellitus in female dogs (68.75%) was significantly (p<0.05) higher than that of male dogs (31.25%). The results of this study showed that the prevalence of diabetes mellitus was higher in old age dogs (>9 years) as compared with other age groups. Diabetes mellitus was found to be more prevalent in winter season same as in human beings. The clinical signs like Polyuria (81.25%), Polydipsia (68.75%), Hyperglycemia (100%), Dehydration (43.75%), Anemia (56.25%) and cataract (18.75%) were commonly observed in diabetic dogs. In the present study the pet dog breed like Yorkshire terrier, Samoyed, Pomuranian and Cross bred were prone to have diabetes mellitus whereas German shepherd, Bull dogs, Pugs and Labrador retriever were at low risk for this malady.
In the present project, the haematological study was conducted on 28 dogs (n=14 diabetic and n=14 healthy dogs). The five ml blood sample were collected from each dog directly from the cephalic vein and were shifted into a sterilized plastic bottle coated with EDTA@1mg/ml and were further analyzed through haemtological analyzer. The results showed the decreased number of RBCs (4.63±0.42) and an increased in PCV values (62.91±4.18). The results also showed dehydration and anemia in diabetic dogs.
In the present study the 21 dogs (n= 14 diabetic dogs, n= 7 healthy dogs) were selected for therapeutic trials. The dogs were divided into further three groups; A, B and C. Each study group was comprised of 7 animals. Group A (comprised of diabetic dogs) was treated with Humulin-N (Insulin) @ 0.5 U/Kg, q12h, SC for 7 days. The group B (comprised of diabetic dogs) was kept as positive control while the healthy dogs were kept in group C for negative control. After the seven days treatment of group A, the blood samples were collected every 2 hr interval for 12 hrs for measuring the glucose value. The process was repeated once in a week for one month. The four blood glucose curves were made. Each blood glucose curve has its own nadir value. The time of glucose nadir value was indicated the peak action of insulin. The blood glucose curves were made to evaluate the effectiveness of insulin. The therapeutic results of this study concluded that the Humulin-N (insulin) was much effective for management of diabetic dogs.
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Therapeutic Trials Against Salmonella Enterica Prevailing In Diarrheic Lambs And Kids
Material type: Book ; Format:
; Literary form:
Publisher: 2014 Dissertation note: Abstract
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Prevalence, Molecular Diagnosis And Chemotherapy Of Degnala Disease In Large Ruminants Of Punjab.
Material type: Book ; Literary form:
Publisher: 2017 Dissertation 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.
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