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

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.

Identification of 11 std Pathogens in Semen Using Polymerase Chain Reaction (pcr) and “Flow-through” Hybridization Technology

“…Caspase activity has been shown to be present in human sperm [33][34][35]. Furthermore, in infertile men a higher percentage of sperm with activated caspases was found, confirming the existence of a caspase-dependent apoptotic pathway in ejaculated human sperm [36,37]. In the third part of our study, we studied the activation of caspase 3 in spermatozoa of infertile men.…”

Mechanisms of the harmful effects of bacterial semen infection on ejaculated human spermatozoa: potential inflammatory markers in semen

“…Satta et al [18] observed sperm PS externalization and DNA fragmentation during experimental C. trachomatis semen infection. Lastly, C. trachomatis infection of semen in vivo has been found to increase mitochondrial depolarization, as well as caspase-3 activation [15]. A significant reduction in ∆ym, viability, and motility was also observed after the incubation of sperm with E. coli and its soluble factors [7].…”

“…In turn, SIF can inhibit sperm motility and viability by decreasing mitochondrial ATPase activity [35]. However, a loss of sperm motility concomitant with the integrity of the sperm mitochondrial membrane potential (∆ym) during experimental in vitro semen infection has also been reported in the presence of bacteria that do not produce SIF, such as known pathogenic C. trachomatis [15] and the conditionally pathogenic S. haemolyticus and B. ureolyticus [9,10]. Most probably, a complex balance between ↓ acrosome reaction (in vitro) [27] [23]…”

Effects ofChlamydia trachomatisinfection on sperm chromatin condensation and DNA integrity

“…This study investigates the role of Chlamydia trachomatis ( C. trachomatis ) infection on sperm chromatin/DNA in male partners of infertile couples. A direct effect of C. trachomatis EBs on spermatozoa that can lead to infertility is premature sperm death (Al‐Mously & Eley, ) through an apoptosis mechanism which is induced by C. trachomatis lipopolysaccharide (LPS; Gallegos et al., ; Ploskonos & Nikolaev, ; Sellami et al., ). The mechanism as defined in different studies involves excessive production of reactive oxygen species (ROS) in leucocytes which has been induced by C. trachomatis lipopolysaccharide (Fraczek & Kurpisz, ) and ROS is mediator of apoptosis (Sellami et al., ; Tafuri, Ciani, Iorio, Esposito, & Cocchia, ).…”