Shakeel Hussain (2007-VA-463)
Molecular Diagnosis Of Babesiosis In Cattle With Special Reference To Cardinal Signs In District Lahore, Punjab - 2015. - 35p.;
Tick infestation and the resulting transmission of serious pathogens in ruminants is one of the most important problems of the livestock industry in developing countries (Aktas et al. 2012).Bovine babesiosis is economically the most important tick-borne disease of cattle worldwide including areas of Australia, Africa, South and Central America. Babesia species are protozoan parasites that parasitize the erythrocytes of domestic animals and humans, causing Anemia in the host affected. Polymerase chain reaction (PCR), which is more sensitive and specific technique, offers an alternative approach for the diagnosis of Babesiosis (Zulfiqar et al. 2012).
Geo-climatic condition of Punjab, Pakistan favours the multiplication and survival of ticks which play a major role in the biological transmission of Tick Born Diseases. In earlier reports the prevalence of cattle tick infestation was more than 50% from Punjab (Durrani et al. 2008, Sajid et al. 2009).
Keeping in views the importance of the disease, the present study was carried out to determine the prevalence of Babesiosis in cattle of Lahore, District of the Punjab, Pakistan. A total of sixty (60) blood samples was collected randomly from dairy cattle of District Lahore. These samples were transported to the Laboratory of Parasitology, Department, University of Veterinary & Animal Sciences, Lahore and were kept at 4oc until further processing for Microscopic examination (Zakir et al. 2014) and then for PCR. We focused on the early detection of Babesiosis through Microscopic examination of Blood samples. For further confirmation of Babesiosis, the blood samples were processed through Polymerase Chain Reaction (PCR) as described by Zulfiqar et al. 2012.
The thick and thin smears of the blood samples were made on the new particularly labeled glass slides. The dried blood smears were fixed in absolute methyl alcohol for one
Summary
32
minute. Staining was performed using Giemsa Stain as method followed by Zakir et al. 2014 i.e. the glass slides bearing thick and thin blood smears were stained with one fourth of dilution of commercially available Giemsa stain for four minutes and were observed under oil immersion at 100X objective to detect the presence of Babesiosis.
All the blood samples were examined through Microscopy showing 04 positive ones, then all the samples were processed using PCR for final confirmation of Babesiosis in Cattle. PCR was performed under the conditions as previously described by Zulfiqar et al. 2012. PCR reaction was performed to obtain amplified products over 30 cycles by 94ºC for 5 min., 94ºC for 30 sec., 50ºC for 30 sec., 72ºC for 45 sec. and completed with a final extension step of 7 min. at 72ºC. Finally the amplified DNA fragments were analyzed after electrophoresis on 1.5% agarose gel.
Prevalence rate will be determined with the help of the following formula:
Prevalence rate = No. of positive samples / No of total samples x 100
Department of Parasitology
2404-T
Molecular Diagnosis Of Babesiosis In Cattle With Special Reference To Cardinal Signs In District Lahore, Punjab - 2015. - 35p.;
Tick infestation and the resulting transmission of serious pathogens in ruminants is one of the most important problems of the livestock industry in developing countries (Aktas et al. 2012).Bovine babesiosis is economically the most important tick-borne disease of cattle worldwide including areas of Australia, Africa, South and Central America. Babesia species are protozoan parasites that parasitize the erythrocytes of domestic animals and humans, causing Anemia in the host affected. Polymerase chain reaction (PCR), which is more sensitive and specific technique, offers an alternative approach for the diagnosis of Babesiosis (Zulfiqar et al. 2012).
Geo-climatic condition of Punjab, Pakistan favours the multiplication and survival of ticks which play a major role in the biological transmission of Tick Born Diseases. In earlier reports the prevalence of cattle tick infestation was more than 50% from Punjab (Durrani et al. 2008, Sajid et al. 2009).
Keeping in views the importance of the disease, the present study was carried out to determine the prevalence of Babesiosis in cattle of Lahore, District of the Punjab, Pakistan. A total of sixty (60) blood samples was collected randomly from dairy cattle of District Lahore. These samples were transported to the Laboratory of Parasitology, Department, University of Veterinary & Animal Sciences, Lahore and were kept at 4oc until further processing for Microscopic examination (Zakir et al. 2014) and then for PCR. We focused on the early detection of Babesiosis through Microscopic examination of Blood samples. For further confirmation of Babesiosis, the blood samples were processed through Polymerase Chain Reaction (PCR) as described by Zulfiqar et al. 2012.
The thick and thin smears of the blood samples were made on the new particularly labeled glass slides. The dried blood smears were fixed in absolute methyl alcohol for one
Summary
32
minute. Staining was performed using Giemsa Stain as method followed by Zakir et al. 2014 i.e. the glass slides bearing thick and thin blood smears were stained with one fourth of dilution of commercially available Giemsa stain for four minutes and were observed under oil immersion at 100X objective to detect the presence of Babesiosis.
All the blood samples were examined through Microscopy showing 04 positive ones, then all the samples were processed using PCR for final confirmation of Babesiosis in Cattle. PCR was performed under the conditions as previously described by Zulfiqar et al. 2012. PCR reaction was performed to obtain amplified products over 30 cycles by 94ºC for 5 min., 94ºC for 30 sec., 50ºC for 30 sec., 72ºC for 45 sec. and completed with a final extension step of 7 min. at 72ºC. Finally the amplified DNA fragments were analyzed after electrophoresis on 1.5% agarose gel.
Prevalence rate will be determined with the help of the following formula:
Prevalence rate = No. of positive samples / No of total samples x 100
Department of Parasitology
2404-T