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Molecular Characterization Of Bovine Viral Diarrhea Virus (Bvdv) In Commercially Available Bovine Semen In Pakistan

By: Muhammad Usman Sajjad khan (2010-VA-223) | Dr. Arfan Ahmad.
Contributor(s): Prof. Dr. Masood Rabbani | Dr. Mati-Ur-Rehman Khan.
Material type: materialTypeLabelBookPublisher: 2017Description: 47p.Subject(s): MicrobiologyDDC classification: 2876-T Dissertation note: The production potential of livestock is showing some compromising trends. This may be due to multiple factors including management, nutrition, stresses, microbes etc. Among microbes, BVD virus has been identified during a study conducted on limited number of samples. The source of BVD virus may be carrier status of imported exotic animals or exotic bovine semen. In Pakistan, most of the farmers are availing services of artificial insemination for breeding purposes without knowing the status of BVD virus. Beside the imported semen, it is also being produced at various semen production units and among others, BVD viruses are the main obstacle in generating revenue by exporting quality semen. These issues are leading to a stage of losing trust of the local clients in purchasing substandard semen on one hand and on the other hand not meeting the export requirements of various client countries. This study aims for the detection and molecular characterization of BVD virus in commercially available bovine semen and the comparative status of BVD virus in the semen of local and exotic breeds of cattle. Furthermore, the comparative status of BVD virus in semen of cattle and buffalo will also be observed. The hypothesis for the current study is that bovine semen used for artificial insemination is a source of BVD virus. In this study, a total of 84 commercial semen straws were procured and processed. Among these, 45 samples (25 Govt. and 20 Private SPUs) originating from 5 breeds of cattle including Dhanni, Dajjal, Sahiwal, Red Sindhi and Cholistani were procured. Similarly, 39 samples (24 from Buffalos and 15 from Exotic Cattle) were processed. All the samples were tested for confirmation of BVD virus through PCR using BVD virus specific primers. Summary 35 The samples if found positive for BVD virus were to be further processed for identification of genotype in semen samples. Furthermore, all the samples were also tested to calculate the bacterial load (TVC/mL) by Plate Method of Colony counting. All the samples were cultured on MH agar, and the colonies were counted after an incubation period of 24 hours at 37ºC The data obtained was analyzed statistically by using Binomial and Chi Square Tests by using the software ‘IBM SPSS Statistics’ (v18) Actual Data about the status and circulating genotype of the BVDV in the commercial bovine semen of local and imported bovines was obtained, along with the status of bacterial contamination of commercial semen available in Pakistan. The study showed that 0% percent of the tested semen samples had detectable BVDV RNA.. The Bacterial contamination of the semen samples was also interesting in the sense that a wide range of Bacterial Load was found in the samples; ranging from 600 CFU/mL to 9250 CFU/mL. Overall the Government SPUs showed a higher contamination with an average Bacterial load of 1975.2 CFU/mL in cattle semen and 3424 CFU/mL in Buffalo semen. In comparison; private SPUs had an average load of 1842 CFU/mL for cattle semen and 2482 CFU/mL for Buffalo semen. Imported semen had an average bacterial load of 1128 CFU/mL
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The production potential of livestock is showing some compromising trends. This may be due to multiple factors including management, nutrition, stresses, microbes etc. Among microbes, BVD virus has been identified during a study conducted on limited number of samples. The source of BVD virus may be carrier status of imported exotic animals or exotic bovine semen. In Pakistan, most of the farmers are availing services of artificial insemination for breeding purposes without knowing the status of BVD virus. Beside the imported semen, it is also being produced at various semen production units and among others, BVD viruses are the main obstacle in generating revenue by exporting quality semen. These issues are leading to a stage of losing trust of the local clients in purchasing substandard semen on one hand and on the other hand not meeting the export requirements of various client countries. This study aims for the detection and molecular characterization of BVD virus in commercially available bovine semen and the comparative status of BVD virus in the semen of local and exotic breeds of cattle. Furthermore, the comparative status of BVD virus in semen of cattle and buffalo will also be observed.
The hypothesis for the current study is that bovine semen used for artificial insemination is a source of BVD virus.
In this study, a total of 84 commercial semen straws were procured and processed. Among these, 45 samples (25 Govt. and 20 Private SPUs) originating from 5 breeds of cattle including Dhanni, Dajjal, Sahiwal, Red Sindhi and Cholistani were procured. Similarly, 39 samples (24 from Buffalos and 15 from Exotic Cattle) were processed. All the samples were tested for confirmation of BVD virus through PCR using BVD virus specific primers.
Summary
35
The samples if found positive for BVD virus were to be further processed for identification of genotype in semen samples. Furthermore, all the samples were also tested to calculate the bacterial load (TVC/mL) by Plate Method of Colony counting. All the samples were cultured on MH agar, and the colonies were counted after an incubation period of 24 hours at 37ºC
The data obtained was analyzed statistically by using Binomial and Chi Square Tests by using the software ‘IBM SPSS Statistics’ (v18)
Actual Data about the status and circulating genotype of the BVDV in the commercial bovine semen of local and imported bovines was obtained, along with the status of bacterial contamination of commercial semen available in Pakistan.
The study showed that 0% percent of the tested semen samples had detectable BVDV RNA..
The Bacterial contamination of the semen samples was also interesting in the sense that a wide range of Bacterial Load was found in the samples; ranging from 600 CFU/mL to 9250 CFU/mL. Overall the Government SPUs showed a higher contamination with an average Bacterial load of 1975.2 CFU/mL in cattle semen and 3424 CFU/mL in Buffalo semen. In comparison; private SPUs had an average load of 1842 CFU/mL for cattle semen and 2482 CFU/mL for Buffalo semen.
Imported semen had an average bacterial load of 1128 CFU/mL

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