Molecular detection of Chlamydia trachomatis and other sexually transmitted bacteria in semen of male partners of infertile couples in Tunisia: the effect on semen parameters and spermatozoa apoptosis markers.

Hanen Sellami,Abir Znazen,Nour Louati,Radhouane Gdoura,Soumaya Ben Zarrouk,Afifa Sellami,Leila Keskes,Hela Mnif,Adnene Hammami,Tarek Rebai,Anil Kumar

doi:10.1371/journal.pone.0098903

Abstract

This study was undertaken to determine the prevalence of Chlamydia trachomatis, Mycoplasmas, and Ureaplasmas in semen samples of the male partners of infertile couples and to investigate whether Chlamydia trachomatis could initiate apoptosis in human spermatozoa. A total of 85 males partners of infertile couples undergoing routine semen analysis according to World Health Organization guidelines were included. Specimens were examined for the presence of Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma hominis, Mycoplasma genitalium, Ureaplasma urealyticum and Ureaplasma parvum by Real time PCR (qPCR). Semen specimens were analysed for the appearance of apoptotic markers (sperm DNA fragmentation, activated caspase 3 levels, mitochondrial membrane potential (ΔΨm)) using flow cytometry. C. trachomatis, N. gonorrhoeae, U. urealyticum, M genitalium were detected in semen samples of 13 (15.2%), 5 (5.8%), 5 (5.8%) and 3 (3.5%) male partners of infertile couples, respectively. M. hominis and U. parvum were detected in semen sample of only one patient (1.1%). The semen of infertile men positive for C. trachomatis showed lower mean of semen count and lower rapid progressive motility (category [a]) of spermatozoa compared to uninfected men with statistically significances (p = 0.02 and p = 0.04, respectively). Flow cytometry analyses demonstrated a significant increase of the mean rate of semen with low ΔΨm and caspase 3 activation of infertile men positive for C. trachomatis compared to uninfected men (p = 0.006 and p = 0.001, respectively). DNA fragmentation was also increased in sperm of infertile men positive for C. trachomatis compared to uninfected men but without statistical significances (p = 0.62). Chlamydial infection was associated to loss of ΔΨm and caspase 3activation. Thus, C. trachomatis infection could be incriminated in apoptosis induction of spermatozoa. These effects may explain the negative direct impact of C. trachomatis infection on sperm fertilizing ability.

Highlights

Transmitted infections are of major concern to researchers and clinicians in the field of reproductive medicine
U. urealyticum, M. genitalium, U. parvum and M. hominis were detected in 5 patients (5.8%), 3 patients (3.5%), 1 patient (1.1%) and 1 patient (1.1%) respectively
The leukocyte count in the male partners of infertile couples with C. trachomatis DNA in semen specimens was higher but not significantly than those uninfected semen (0.86106/ml vs 0.46106/ml, p = 0.36) (Table 3)

Summary

Introduction

Transmitted infections are of major concern to researchers and clinicians in the field of reproductive medicine. It is estimated that 15% of male infertility is related to genital tract infection [1]. According to a World Health Organization (WHO) [3] report, C. trachomatis is responsible for the most common sexually transmitted bacterial infection worldwide, affecting more than 90 million people and has been known for some time to have a significant effect on human reproduction [4]. Some reports indicated that C. trachomatis infection is associated with a decrease in sperm concentration and motility and with altered semen pH and reduced volume of the ejaculate [11,12,13,14]. Other studies have revealed no association between C. trachomatis infection of the male genital tract and altered sperm quality [9,15,16,17,18,19,20,21]. The available evidence is conflicting and still makes it impossible to establish a clear relationship between C. trachomatis infection and semen quality

Objectives

Methods

Results

Conclusion