Pseudomonas aeruginosa: A risk factor for fertility in male mice.

The present study was designed to evaluate the effect of P. aeruginosa on reproductive potential of male mice via a series of in vitro and in vivo experiments. In vitro studies involved sperm parameters, Mg2+ATPase activity and acrosome status. In vivo study employed male mice which in the right vas deferens received 20 µl of either PBS (Group I) or 104 cfu of P. aeruginosa (Group II). The animals were sacrificed on day 3, 7 and 14 and various parameters viz. body weight, TSI (%), bacterial load, spermiogram {i.e. sperm count, motility (%), viability (%) and morphology}, lipid peroxidation and tissue histopathology were evaluated. The results revealed that cell free supernatant of P. aeruginosa resulted in reduced motility, viability, Mg2+dependent ATPase activity and premature acrosomal loss of mouse spermatozoa in vitro. In vivo study showed that in comparison to group I, group II revealed significant alterations in all the parameters on all the days of sacrifice. Further, when reproductive organs of right and left side of mice in group II were compared, the right side demonstrated more devastating effects in terms of altered TSI (%) of testis and cauda epididymis, higher bacterial counts, azoospermia, increased malondialdehyde levels and severe inflammation in tissue histopathology in comparison to left side where bacteria disseminated in reduced numbers, thereby, resulting in insignificant changes in TSI (%), spermiogram, malondialdehyde levels and tissue histology. This study demonstrates that the colonization of P. aeruginosa in male genital tract could be a risk factor for fertility.

Effect of Mycoplasma bovis and Mycoplasma bovigenitalium in semen on fertilization and association with in vitro produced morula and blastocyst stage embryos.

Frozen-thawed bovine semen contaminated with Mycoplasma bovis (M. bovis) or Mycoplasma bovigenitalium (M. bovigenitalium) at either a high (10(6) CFU/mL) or low (10(4) CFU/mL) concentration was used for bovine oocyte insemination. The resulting embryos were washed 10 times as recommended by the International Embryo Transfer Society (IETS) prior to isolation of agent. A total of 1494 oocytes was inseminated with contaminated sperm cells and 855 oocytes with uninfected control semen. There was a significantly higher proportion of embryos that developed to the blastocyst stage in control than in the mycoplasma exposed groups (P<0.05). Isolation of motile spermatozoa by swim-up procedure prior to insemination did not render sperm cells free of Mycoplasma spp. Although M. bovis was isolated from all washed embryos after the high exposure level, it was found in only 60% of the samples after the low exposure level. In contrast, M. bovigenitalium was isolated from 70 and 12% of washed embryos exposed to the high and low levels of microorganism, respectively. Using scanning electron microscopy, both microorganisms were detected in association with the surface of zona pellucida-intact embryos and with sperm cells. These results indicate that mycoplasmas present in semen can be transmitted through the IVF system and infect embryos. Furthermore, the experiments showed that supplementation of culture media with standard antibiotics and washing embryos as recommended by IETS were not effective in rendering IVF embryos free from M. bovis and M. bovigenitalium.