Antibiotics supplemented culture media can eliminate non-specific bacteria from human semen during sperm preparation for intra uterine insemination

Dissanayake, D.M.A.B.; Amaranath, KA; Perera, R. R. D. P.; Wijesinghe, P.S.

doi:10.4103/0974-1208.130859

Cited by 9 publications

(10 citation statements)

References 19 publications

(25 reference statements)

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“…Presence of these genera in IVF culture media was not previously published. At the same time previous studies have indicated that coliform bacteria, including E. coli that belong to Alphaproteobacteria were found in higher concentration in semen and media used during IVF procedure [5,21,43]. Kala et al demonstrated that inoculation of E. coli caused adhesion to the sperm membrane and subsequent destruction leading to reduced motility and viability in washed samples [44].…”

Section: Discussionmentioning

confidence: 99%

The complex microbiome from native semen to embryo culture environment in human in vitro fertilization procedure

Štšepetova

Baranova

Simm

et al. 2020

Reprod Biol Endocrinol

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Background: Only a few microbial studies have conducted in IVF (in vitro fertilization), showing the high-variety bacterial contamination of IVF culture media to cause damage to or even loss of cultured oocytes and embryos. We aimed to determine the prevalence and counts of bacteria in IVF samples, and to associate them with clinical outcome. Methods: The studied samples from 50 infertile couples included: raw (n = 48), processed (n = 49) and incubated (n = 50) sperm samples, and IVF culture media (n = 50). The full microbiome was analyzed by 454 pyrosequencing and quantitative analysis by real-time quantitative PCR. Descriptive statistics, t-, Mann-Whitney tests and Spearman's correlation were used for comparison of studied groups. Results: The study involved normozoospermic men. Normal vaginal microbiota was present in 72.0% of female partners, while intermediate microbiota and bacterial vaginosis were diagnosed in 12.0 and 16.0%, respectively. The decreasing bacterial loads were found in raw (35.5%), processed (12.0%) and sperm samples used for oocyte insemination (4.0%), and in 8.0% of IVF culture media. The most abundant genera of bacteria in native semen and IVF culture media were Lactobacillus, while in other samples Alphaproteobacteria prevailed. Staphylococcus sp. was found only in semen from patients with inflammation. Phylum Bacteroidetes was in negative correlation with sperm motility and Alphaproteobacteria with high-quality IVF embryos. Conclusion: Our study demonstrates that IVF does not occur in a sterile environment. The prevalent bacteria include classes Bacilli in raw semen and IVF culture media, Clostridia in processed and Bacteroidia in sperm samples used for insemination. The presence of Staphylococcus sp. and Alphaproteobacteria associated with clinical outcomes, like sperm and embryo quality.

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Section: Discussionmentioning

confidence: 99%

The complex microbiome from native semen to embryo culture environment in human in vitro fertilization procedure

Štšepetova

Baranova

Simm

et al. 2020

Reprod Biol Endocrinol

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“…These particular antibiotics were chosen because they may be used in initial semendiluting solutions and are classically associated with resistance development. 5 Of note, though not reaching statistical significance, there was an increase of resistance to penicillin among the Staphylococcus isolates, reflecting the increasing incidence of methicillin-resistant Staphylococcus aureus. It is estimated that colonization rates of S. aureus may approach 20% in the United States and that 1% to 3% of the population is colonized with methicillin-resistant S. aureus.…”

Section: Discussionmentioning

confidence: 98%

“…4 Reports of semen samples positive for bacteria are extremely variable, with rates of 45% to 100%. 5 Longitudinal studies of the variation of these bacteria over time as well as their resistance patterns have not been commonly reported. At our institution, to ensure appropriate processing of semen, residues from semen samples are routinely evaluated for bacteria with antibiotic sensitivity profiles.…”

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confidence: 99%

Time trends for bacterial species and resistance patterns in semen in patients undergoing evaluation for male infertility

Machen

Bird

Brown

et al. 2018

Baylor University Medical Center Proceedings

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Semen from asymptomatic men who are being evaluated as male partners in interfile couples have been reported to contain a variety of bacteria. Longitudinal studies of the variation of these bacteria over time and their resistance patterns have not been commonly reported. At our institution, residues from semen samples are routinely evaluated for bacteria, including antibiotic sensitivity profiles. We set out to profile the changes in semen bacteria and antibiotic resistance at our institution over time. A total of 72 semen isolates were examined for type of bacteria and sensitivity to a panel of antibiotics. The results were divided into two separate 5-year intervals (the first beginning in 2006, the second in 2011) and compared. The majority of bacteria were skin flora, with Streptococcus and Staphylococcus being the most prevalent. The resistance data for these two pathogens showed minimal statistically significant difference between the two time periods, although the Staphylococcus species did show a trend toward increasing resistance, suggesting that antibiotics currently used in sperm cell preparations may need to be varied.KEYWORDS Bacterial resistance; infertility; male; semen; semen analysis S emen from asymptomatic men being evaluated for infertility has been reported to contain a variety of bacteria. Frequently, these results are from contamination, the most likely sources of which are the male urethra or skin secondary to improper collection or colonization.1,2 Further, incidentally identified semen bacteria in the absence of symptoms are typically considered to have limited, if any, clinical significance.1,3 These species vary by population, but the most common bacteria are typically gram-positive organisms, such as Staphylococcus or Streptococcus species. 4 Reports of semen samples positive for bacteria are extremely variable, with rates of 45% to 100%. 5Longitudinal studies of the variation of these bacteria over time as well as their resistance patterns have not been commonly reported. At our institution, to ensure appropriate processing of semen, residues from semen samples are routinely evaluated for bacteria with antibiotic sensitivity profiles. We set out to profile the changes in semen bacteria and antibiotic resistance at our institution over time. METHODSAfter institutional review board approval was obtained, a retrospective review was undertaken over a 10-year period from 2006 to 2016. Approximately every 2 months, the residues of semen samples collected were sent for microbiology evaluation. The samples were typically leftover portions from freshly collected specimens of men undergoing fertility evaluations. Men were advised to abstain from intercourse 3 to 5 days prior to collection, wash their hands and genitals thoroughly before sample production, and produce the sample into a sterile specimen cup. Isolates were then examined for sensitivity to a panel of antibiotics. The results were divided into two separate 5-year intervals (the first beginning in 2006, the second in 2011) a...

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“…Different bacterial species have been isolated during storage in liquid nitrogen, such as Stenotrophomonas maltophilia, Bacillus spp., Enterobacter spp., and Staphylococcus spp., some of them related to the suppression of embryonic development [4,6]. Thus, to minimise the risks of microbial growth, antibiotics have been introduced at sample collection, fertilisation, culture, or storage [17][18][19]. However, the opposite effect of antibiotics on the growth rate of preimplantation embryos has been well documented [19][20][21].…”

Section: Introductionmentioning

confidence: 99%

“…Thus, to minimise the risks of microbial growth, antibiotics have been introduced at sample collection, fertilisation, culture, or storage [17][18][19]. However, the opposite effect of antibiotics on the growth rate of preimplantation embryos has been well documented [19][20][21]. Taking all these data into account, the bacterial cross-infection risk in embryo cryopreservation is deemed a significant public health concern [22].…”

Section: Introductionmentioning

confidence: 99%

Experimental Evidence Reveals Both Cross-Infection and Cross-Contamination Risk of Embryo Storage in Liquid Nitrogen Biobanks

Marín

García‐Domínguez

Montoro-Dasi

et al. 2020

Animals

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In recent decades, gamete and embryo cryopreservation have become routine procedures in livestock and human assisted reproduction. However, the safe storage of germplasm and the prevention of disease transmission continue to be potential hazards of disease transmission through embryo transfer. This study aimed to demonstrate the potential risk of cross-infection of embryos from contaminated liquid nitrogen, and cross-contamination of sterile liquid nitrogen from infected embryos in naked and closed devices. Additionally, we examined the effects of antibiotic-free media on culture development of infected embryos. The study was a laboratory-based analysis using rabbit as a model. Two experiments were performed to evaluate both cross-infection (liquid nitrogen to embryos) and cross-contamination (embryos to liquid nitrogen) of artificially inoculated Salmonella Typhimurium, Staphylococcus aureus, Enterobacter aerogenes, and Aspergillus brasiliensis. Rapid cooling through vitrification was conducted on rabbit embryos, stored for a year, thawed, and cultured. In vivo produced late morulae–early blastocyst stages (72 h) embryos were used (n = 480). Embryos were cultured for 1 h in solutions with and without pathogens. Then, the embryos were vitrified and stored in naked and closed devices for one year in two liquid nitrogen biobanks (one pathogen-free and the other artificially contaminated). Embryos were warmed and cultured for a further 48 h, assessing the development and the presence of microorganism (chromogenic media, scanning electron microscopy). Embryos stored in naked devices in artificially contaminated liquid nitrogen became infected (12.5%), while none of the embryos stored in closed devices were infected. Meanwhile, storage of artificially infected embryos incurred liquid nitrogen biobank contamination (100%). Observations by scanning electron microscopy revealed that all the microorganisms were caught in the surface of embryos after the vitrification-thawed procedure. Nevertheless, embryos cultured in antibiotics and antimycotic medium developed to the hatched blastocyst stage, while artificially infected embryos cultured in antibiotic-free medium failed to develop. In conclusion, our findings support that both cross-contamination and cross-infection during embryo storage in liquid nitrogen biobanks are plausible. So, to ensure biosafety for the cryogenic storage, closed systems that avoid direct contact with liquid nitrogen must be used. Moreover, it seems essential to provide best practice guidelines for the cryogenic preservation and storage of gametes and embryos, to define appropriate quality and risk management procedures.

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Antibiotics supplemented culture media can eliminate non-specific bacteria from human semen during sperm preparation for intra uterine insemination

Cited by 9 publications

References 19 publications

The complex microbiome from native semen to embryo culture environment in human in vitro fertilization procedure

The complex microbiome from native semen to embryo culture environment in human in vitro fertilization procedure

Time trends for bacterial species and resistance patterns in semen in patients undergoing evaluation for male infertility

Experimental Evidence Reveals Both Cross-Infection and Cross-Contamination Risk of Embryo Storage in Liquid Nitrogen Biobanks

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