Bacteria in semen used for IVF affect embryo viability but can be removed by stripping cumulus cells by vortexing.
Bacterial contamination of in vitro vs in vivo produced embryos presents a particular danger because of the alteration of the zona pellucida and the use of various biological products during culture. Our objective was to investigate the effects of semen contaminated with bacteria on IVF of bovine oocytes and to determine if removal of cumulus cells by vortexing as opposed to pipetting would reduce contamination and improve subsequent embryonic development. Semen from 5 bulls of the Native Korean breed (Bulls A, B, C, D, E) was used for IVF of matured oocytes. Preliminary studies had shown that the semen from Bulls A, B, D and E but not Bull C was contaminated with various species of common bacteria. After IVF, the cumulus cells surrounding the oocytes were removed either by pipetting or vortexing. Viability and cleavage rates of the resulting zygotes was assessed after 44 h in culture. When cumulus cells were removed by pipetting, only zygotes derived from oocytes that were fertilized with uncontaminated semen from Bull C developed to morula and blastocyst stages; zygotes derived from oocytes that were fertilized with contaminated semen from Bulls A, B, D and E started to degenerate, and the culture media became noticeably turbid. When cumulus cells were removed by vortexing, zygotes derived from oocytes fertilized with either contaminated or uncontaminated semen showed good rates of development (16 to 32%) to morula or blastocyst stages. From these results it can be concluded that the bacteria introduced with the semen contaminated the in vitro system and severely reduced the viability of the embryos. In contrast, complete removal of the cumulus cells with vortexing, as opposed to pipetting, reduced the contamination of the culture medium, allowing embryonic development to take place.
Kim
I H
IH
National Livestock Research Institute, RDA, Cheonan, Korea.
Son
D S
DS
Lee
H J
HJ
Yang
B C
BC
Lee
D W
DW
Suh
G H
GH
Lee
K W
KW
Jung
S C
SC
D000818:Animals; D002417:Cattle / Q000196:embryology*; D002469:Cell Separation / Q000379:methods / Q000662:veterinary*; D002498:Centrifugation / Q000662:veterinary; D005314:Embryonic and Fetal Development; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D008297:Male; D009865:Oocytes / Q000382:microbiology*; D012661:Semen / Q000382:microbiology*; D012662:Semen Preservation / Q000662:veterinary; D013084:Sperm-Ovum Interactions; D015044:Zona Pellucida / Q000382:microbiology