157 VIRAL RISKASSESSMENT OF BOVINE OOCYTES HARVESTED FROM ABATTOIR ORIGIN

Abstract

Bovine oocytes derived from abattoir origin are mainly used for artificial reproductive techniques in both agricultural and biomedical applications. Regulatory agencies have expressed concern for potential transmission of adventitious viruses by sourcing bovine oocytes from abattoir origin. To evaluate this concern, a viral risk assessment was conducted on batch samples collected from follicular fluid, cumulus cells, oocytes, and Day 8 embryos. These batch samples were collected from ovaries on seven randomly selected days in a 2-week period and they were frozen and stored at –80°C until tested. All samples were tested by 9 Code of Federal Regulations (9CFR) part 113.53c (animal viral testing) at a GLP compliant laboratory (American BioResearch Laboratories, Sevier, TN, USA). The 9CFR viral testing includes bovine viral diarrhea virus (BVDV), bovine parvovirus, bovine adenovirus type 3 and 5, bovine rabies virus, bovine bluetongue virus, bovine respiratory syncytial virus, bovine reovirus, viral cytopathic effect, and hemadsorption on permissive cell cultures. Batch samples were also tested for BVDV and bovine leukemia virus (BLV) by polymerase chain reaction (PCR) and follicular fluids for BVDV antibody neutralization activity at an accredited diagnostic laboratory (Animal Disease Research and Diagnostic Laboratory, SDSU, Brookings, SD, USA). The 9CFR viral testing results on all the batch samples were negative. The BVDV PCR test had a low positive (37.89 cycles) with one follicular fluid batch sample (1/7) and a low positive (37.9 cycles) on all cleaved embryos (7/7). However, BVDV virus isolation was negative for both batch samples by 9CFR testing. The BLV PCR had a positive follicular batch sample (1/7), with all other samples testing being negative. BVD serum neutralization antibody assay demonstrated that all follicular fluid had significant titers of 1:128–1:1024. Although some of the viral particles may have been detected in follicular fluid and cleaved embryos by PCR, none of the batch samples collected were positive for viral growth in this study. The BVDV PCR indicated low levels of BVDV RNA. It is speculated that the positive BVDV PCR results on cleaved embryos could possibly be attributed to the use of irradiated fetal calf serum (contaminated BVDV virus) used in culture media. Follicular fluids also have high titers to BVDV which may have neutralized the virus. These results indicate that virus-free bovine oocytes can be derived from the abattoir. Thus, with appropriately applied quality assurance testing, abattoir-origin oocytes might be used effectively in agricultural and biomedical applications.

Table 1. Summary of test results on batch samples from abattoir-derived oocytes