Microbiological profiles of Semen culture and correlations with sperm parameters and sperm DNA integrity

Infertility is the condition characterised by the failure to conceive a pregnancy after engaging in regular sexual intercourse for one year. Around 15% of couples in their reproductive years are unable to conceive a desired pregnancy within a 12-month timeframe, even with consistent unprotected sexual intercourse. Congenital and acquired abnormalities in the urogenital system, male accessory gland infections (MAGI), elevated scrotal temperature, disruptions in hormone levels, genetic abnormalities, and immunological factors can cause male infertility. Inflammatory processes and infections are prominent contributors to infertility. In the majority of infertile couples, the primary cause of male infertility is aberrant semen parameters. It has a crucial impact on reproductive results, independently or in conjunction with female variables. Semen culture is a crucial diagnostic method for evaluating infections in the genitourinary tract. Bacteria in seminal fluid, known as bacteriospermia, is considered a clinical indication of an active illness. Individuals' urinary systems cause semen contamination, which can spread sexually through infections. Male infertility is considered a serious healthcare issue that has a significant negative impact on male reproductive ability. Around 15% of infertile men exhibit a substantial presence of bacterial infections in their semen. The infection's potential consequences may include impaired sperm production, function, and transport. This prospective case-control study included 92 males suffering from infertility, and 30 healthy males served as a control group. After three days of abstinence, we collected the semen specimens from both groups using a disposable sterile seminal fluid container. Once we completed the liquefaction of the seminal fluid, we divided the semen specimens into three volumes. The first volume was used for seminal analysis to determine semen parameters, the second volume was used for bacteriological culture, and the last was used for DNA fragmentation. All semen specimens were cultured in blood agar, chocolate agar, and MacConkey agar, and then, after being obtained for pure culture, the pure colony from each culture was used for the identification of bacterial isolates using the analytical profile index (API). The sperm DNA fragmentation was detected using an alanine blue stain (ABS). According to the age distribution, this study showed that the highest number of patients, 56 (60.8%), were within the age range of 30-39 years, followed by iv 25 (27.2%) within the age group <30 years. In addition, the lowest number of patients, 11 (12%), were within the age group of ≥40 years. Regarding lifestyle variables, 55 (59.8%) were current smokers, while 37 (40.2%) were nonsmokers. 49 (53.3%) of infertile males were abstinence for ≤ 3 days; in contrast, 43 (46.7%) of them were abstinence for >3 days. Furthermore, 60 (65.2%) of infertile males had <3 years of nonfertilization period, and 23 (25%) had 3-5 years of non-fertilization period, while the lower number of infertile males 9 (9.8%) had more than 5 years of non-fertilization period. When looking at the bacteria in the semen culture, Enterococcus faecalis was found 31 times (33.7%), then Escherichia coli 25 times (27.2 %), Streptococcus agalactiae 14 times (15.2%), Staphylococcus aureus 11 times (% 12), and Proteus species 6 times (6.5%). Klebsiella pneumonia was found 5 times (5.4 %). In terms of the semen parameters, there was a statistically significant difference between the bacterial isolates in terms of semen volume, progression, immobility (p-value <0.0001), and DNA fragmentation (p-value 0.0063). On the other hand, there was no statistically significant difference between the bacterial isolates in terms of semen concentration (pvalue 0.424), and total count (p-value 0.366), but there is statistically significant difference in semen non-progression (p-value 0.027). Additionally, there was a significant difference in the amount of semen between the two study groups. Infections with S. aureus had the most significant effect (p-value 0.0008), followed by infections with K. pneumonia (p-value 0.0093) and E. coli (p-value 0.003). Infections with E. faecalis had the least significant effect (p-value 0.0188). In comparison to healthy controls, there was a significant effect on the seminal concentration of infertile males (p-value <0.0001 for S. agalactiae, E. coli, S. faecalis, S. aureus, and Proteus spp., and p-value 0.0003 for K. pneumonia). Regarding total sperm count, S. faecalis, E. coli, and S. agalactiae infections had a high statistically significant impact (p-value <0.0001), whereas S. aureus infection (p-value 0.0005) and Proteus spp. had a slightly lower impact (p-value 0.0017). In contrast, there was no impact on the total count between both groups in the K. pneumonia infection (p-value 0.0596). Additionally, all bacterial isolate infections had a significant impact on sperm non-progression motility among infertile males when compared with healthy controls (p-value <0.0001 for all isolates and p-value 0.003 for K. pneumonia). Moreover, all bacterial isolate infections had a comparable and noteworthy effect on the integrity of sperm, progression, immobility, and DNA in infertile males as compared to healthy controls (p-value <0.0001). The data obtained have shown that infection in the genital tract may play an important role in infertility by affecting sperm quality parameters and DNA integrity.