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Molecular Diagnosis And Therapeutic Trials On Fasciolosis In Sheep And Goats

By: Khalid Mehmood | Dr. Muhammad Ijaz.
Contributor(s): Prof. Dr. Muhammad Sarwar Khan.
Material type: materialTypeLabelBookPublisher: 2012Subject(s): Department of Clinical Medicine & SurgeryDDC classification: 1434,T 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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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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