Removal of bacterial contaminants from semen for in vitro fertilization or artificial insemination by the use of buoyant density centrifugation.
Buoyant density centrifugation of semen produces the accumulation of populations of highly motile, morphologically normal spermatozoa in the lowermost 1 ml of Percoll (Pharmacia Fine Chemicals AB, Uppsala, Sweden) density gradients. In addition, the majority of bacteria present in semen are retained in the seminal plasma at the top of the gradients. Of 40 semen samples examined, 37 contained detectable bacteria, but after buoyant density centrifugation, the spermatozoal populations collected from the lowermost 1 ml of the Percoll columns were found to contain few or no bacteria. When preparations were collected using sterile technique (by boring a hole through the bottom of the centrifuge tube), 14 of the 20 preparations were found to be bacteria-free. When preparations were collected by passing a spinal needle from the surface through the seminal plasma to the bottom of the centrifuge tube, the sterility of the final spermatozoa preparations was not maintained, with only 5 of the 20 samples completely free of bacteria. The residual bacterial contamination of the remaining 15 samples was, however, very low (less than 5 colonies after a 48-hour culture period).
D001419:Bacteria / Q000302:isolation & purification*; D001431:Bacteriological Techniques; D002499:Centrifugation, Density Gradient; D005260:Female; D005307:Fertilization in Vitro; D006801:Humans; D007315:Insemination, Artificial; D008297:Male; D012661:Semen / Q000382:microbiology*