title: Bovine viral diarrhea virus (BVDV) and anti-BVDV antibodies in pooled samples of follicular fluid.
abstract: Bovine viral diarrhea virus (BVDV) can be found in cells and fluids from ovaries collected at the abattoir. On the other hand, immunoglobulins are also found in the fluid of ovarian follicles. Anti-BVDV antibodies in follicular fluid might reduce cross-contamination of COCs at the time of collection or hinder the use of virus isolation to test for the presence of virus. One objective of this study was to determine the frequency with which BVDV could be found in pooled follicular fluid collected during the periodic aspiration of COCs from abattoir-origin ovaries. A second objective was to determine the prevalence and neutralizing activity of anti-BVDV antibodies in these blended samples. We collected samples of pooled follicular fluid (n = 55) over a 20-month period as part of our routine oocyte collection activities. We assayed each sample for BVDV using virus isolation as well as reverse transcription nested polymerase chain reaction (RT-nPCR) procedures. We also tested follicular fluid for antibody that would neutralize four representative strains of BVDV (SD-1, a genotype 1a strain; NY-1, a genotype lb strain; CD-87, a genotype 2 strain, and PA-131, a divergent genotype 2 strain). We detected no BVDV by virus isolation, but we did identify the virus by RT-nPCR in one of the 55 samples of follicular fluid. Automated dye terminator nucleotide sequencing of the amplified portion of the viral genome indicated a genotype 1 strain that was distinct from any of our laboratory strains. In addition, each of the samples of follicular fluid contained sufficient antibody to neutralize large quantities of each of the four laboratory strains that were used. Finding BVDV in just 1 of 55 samples was consistent with reports of similar studies in which the occurrence of BVDV in abattoir-origin materials ranged from 0.9 to 12%. We presumed that failure to isolate the virus was due to neutralizing antibody in the sample. Thus, the incidence of BVDV contamination of our IVF system at the level of pooling of follicular fluid was low for the 20-month period. The presence of anti-BVDV antibody in pooled follicular fluid provided a coincidental means of neutralizing BVDV when it was introduced in fluid aspirated from infected ovaries.
authors.affiliation: College of Veterinary Medicine, Auburn University, AL 36849, USA. galikpk@vetmed.auburn.edu
authors.forename: Patricia K
authors.initials: PK
authors.lastname: Galik
authors.forename: M Daniel
authors.initials: MD
authors.lastname: Givens
authors.forename: David A
authors.initials: DA
authors.lastname: Stringfellow
authors.forename: Elizabeth G
authors.initials: EG
authors.lastname: Crichton
authors.forename: Michael D
authors.initials: MD
authors.lastname: Bishop
authors.forename: Ken J
authors.initials: KJ
authors.lastname: Eilertsen
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis*; D002417:Cattle / Q000821:virology*; D001908:Diarrhea Viruses, Bovine Viral / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D005260:Female; D015571:Follicular Fluid / Q000821:virology*; D005838:Genotype; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000914:Antibodies, Viral