Evaluation and management of male genital tract infections in the setting of male infertility: an updated review.

PURPOSE OF REVIEW: Male infertility may be secondary to male genital tract infection (MGTI) in an estimated 15% of cases. In the absence of overt clinical signs, evaluation for MGTI beyond semen analysis is not well established. Therefore, we review the literature on the evaluation and management of MGTI in the setting of male infertility. RECENT FINDINGS: A set of international guidelines recommends semen culture and PCR testing, but the significance of positive results remains unclear. Clinical trials evaluating anti-inflammatory or antibiotic treatment report improvements in sperm parameters and leukocytospermia, but data on the effect on conception rates are lacking. Human papillomavirus (HPV) and the novel coronavirus (SARS-CoV-2) have been associated with poor semen parameters and decreased conception rates. SUMMARY: The finding of leukocytospermia on semen analysis prompts further evaluation for MGTI, including focused physical examination. The role of routine semen culture is controversial. Treatment options include anti-inflammatories; frequent ejaculation; and antibiotics, which should not be used in the absence of symptoms or microbiological infection. SARS-CoV-2 represents a subacute threat to fertility that should be screened for in the reproductive history along with HPV and other viruses.

Penicillin and amikacin mixture has bactericidal activity equivalent to gentamicin, tylosin, lincomycin and spectinomycin mixture in equine frozen semen.

Neat stallion semen can contain a variety of microorganisms, some of which may impair sperm quality and/or cause infection of the mares' reproductive tract. For this reason, antibiotics are commonly added to semen extenders. A combination of gentamicin, tylosin, lincomycin and spectinomycin (GTLS) has been recommended for use, but there are no reports on the use of this mixture in equine semen extender. Penicillin and amikacin (PA) are safe for preserving sperm quality while effectively controlling bacterial growth in equine cooled stored semen, but data on frozen semen are scarce. Therefore, a bioequivalence study was performed to assess the bactericidal activity of GTLS and PA in equine frozen semen. Nine mature, healthy stallions were used in the study. Split ejaculates were processed using media without antibiotics (Control) or with different antibiotics. For the GTLS group, centrifugation medium and freezing extender were prepared with gentamicin 250 µg/ml, tylosin 50 µg/ml, lincomycin 150 µg/ml and spectinomycin 300 µg/ml. For the PA group, the centrifugation medium was prepared with potassium penicillin G (PPG) 1200 units/ml and the freezing extender was prepared with PPG 1200 units/ml and amikacin 500 µg/ml. Semen processed in extenders without antibiotics had higher (p < .005) bacterial loads throughout all cryopreservation processing steps than semen samples processed using antibiotics. There were no differences in semen bacterial load after centrifugation, 15 and 30 min after final extension, and after thawing between GTLS and PA groups, but PA had faster (p < .05) kill-time kinetics than GTLS. Only minor differences in sperm kinetic parameters were observed among groups. In conclusion, this study demonstrated bioequivalence between GTLS and PA in mitigating end-point bacterial loads. Prudent concentrations of the antibiotic mixtures evaluated in this study can be considered both effective and sperm-safe for equine frozen semen.

Antichlamydia antibodies and sperm quality among male partners of infertile couples ins Nigeria.

Background: The impact of Chlamydia trachomatis on semen quality has been studied with varied results. Aim: To determine the prevalence of antichlamydial antibodies and their relationship with sperm quality among male partners of infertile couples in Enugu, South-East Nigeria. Materials and Methods: It was a cross-sectional study of infertile male partners of couples attending infertility clinics at the University of Nigeria Teaching Hospital (UNTH) Ituku-Ozalla, Enugu, Nigeria. Their sera were assayed for antichlamydial antibodies, and semen analysis and culture were done for each participant. Results: Two hundred and eighty-two (282) male partners of infertile couples were studied. Infertility was commoner among participants aged 40 years or more (45.1%) and was mainly of the "primary type" (62.1%). Antichlamydia antibody was detected in 156 (55.3%) participants and was significantly associated with sperm quality (P = 002; OR = 2.294; 95% CI = 1.36-3.88). Overall, 81 (28.7%) had abnormal sperm quality. The sperm count, progressive motility, and vitality were significantly lower in participants with abnormal sperm quality than those with normal sperm quality (P < 0.001) while morphology, volume, and liquefaction time did not differ significantly (P > 0.05). Staphylococcus aureus was the predominant organism isolated from culture (122/282, 43.3%) while Streptococcus species were the least (4/262, 1.4%). There was significantly more Staphylococcus aureus isolated from the semen of participants that were seropositive to antichlamydial antibodies than those that were seronegative (80/156, 51.3% vs. 42/126, 33.3%; OR = 2.105; 95% CI = 1.30-3.42; P = 0.003). Conclusion: The prevalence of antichlamydial antibodies among male partners of infertile couples in Enugu, Nigeria is high and there is a significant association with sperm quality, sperm count, and bacterial isolates in seminal culture. Male partners of infertile couples in Enugu should be screened for antichlamydial antibodies and appropriate treatment offered wherever indicated. There is a need for increased public awareness and advocacy campaigns on the impact of Chlamydia infection on male factor infertility. This primary preventive measure may help in reducing the burden of Chlamydia infection and male factor infertility.

In vitro transmission of Chlamydia using naturally infected koala (Phascolarctos cinereus) semen.

Transmission of Chlamydia pecorum infection has generally been assumed to be via the urogenital route and in an attempt to confirm this we investigated an in vitro method of Chlamydia infection using naturally infected koala semen to inoculate a cell line and attempt to estimate C. pecorum infectious load. A total of 57% of 122 koala semen samples had low C. pecorum copy number or no burden, while 18% of semen samples contained >10000 inclusion-forming units/mL, as determined by quantitative polymerase chain reaction. In vitro inoculation of a McCoy cell line resulted in successful infection from 4% of semen samples where C. pecorum burden was >105 inclusion-forming units/mL. Our preliminary study suggests that transmission of C. pecorum infectious dose may be restricted to peak bacterial shedding in semen associated with recent infection. Here, we report venereal transmission of C. pecorum in koala semen is possible; however, we speculate that antimicrobial factors and innate immune function receptors associated with semen may inhibit chlamydial growth. These mechanisms have yet to be reported in marsupial semen.

Role of Infection and Leukocytes in Male Infertility.

Male infertility is considered as a multifactorial complex reproductive illness, and male urogenital infection and inflammation are crucial etiologies contributing up to 35% of all cases. Mostly triggered by sexually transmitted diseases and uropathogens, chronic manifestation of such infection may cause irreversible infertility in the male. Male urogenital infection involves bacterial, viral, protozoal, and fungal infections many of which remain asymptomatic most of the time and are passed to the sexual partner leading to fertilization failure, pregnancy loss, and even development of illness in the offspring. The abundance of leukocytes in semen can be used as an indicator of urogenital infection. Its contribution in male infertility can be as high as 30% and the clinical condition is referred to as leukocytospermia. Seminal bacterial load together with increased leukocytes contribute to the impairment of male fertility parameters such as, sperm motility, DNA integrity, acrosome reaction, and damage sperm molecular structure. Pathophysiology of bacteriospermia-induced impairment of male infertility is probably mediated by the involvement of bacterial pathogens in the intrinsic apoptotic pathway resulting in sperm death, whereas that of seminal leukocytes operates through excessive generation of ROS. Although the application of antibiotics forms the frontline therapeutic approach, the growing resistance to antibiotics poses a concern in the management of microbes-induced male urogenital infection. Complementary and alternative medicine may offer additional management options in combating such infections. On the other hand, both broad spectrum antibiotics and antioxidant therapy have showed promising results in the management of infertile men with leukocytospermia. Use of herbal medicine may also play a promising role in the management of such patients. However, recent molecular biology techniques have noted the association of elevated levels of IL-8 with both the Chlamydial infection of the male urogenital tract as well as the clinical condition of leukocytospermia. On the basis of such common pathogenesis, further research involving advanced molecular techniques may pave the way towards the development of better diagnostic tools in the clinical management of male urogenital infection and leukocytospermia.

Sexually transmitted infections and semen quality from subfertile men with and without leukocytospermia.

BACKGROUND: The role of sexually transmitted infections (STIs) in semen parameters and male infertility is still a controversial area. Previous studies have found bacterial infection in a minority of infertile leukocytospermic males. This study aims to investigate the prevalence of STIs in semen from subfertile men with leukocytospermia (LCS) and without leukocytospermia (non-LCS) and their associations with sperm quality. METHODS: Semen samples were collected from 195 men who asked for a fertility evaluation. Infection with the above 6 pathogens was assessed in each sample. Sperm quality was compared in subfertile men with and without LCS. RESULTS: The LCS group had significantly decreased semen volume, sperm concentration, progressive motility, total motility and normal morphology. The infection rates of Ureaplasma urealyticum (Uuu), Ureaplasma parvum (Uup), Mycoplasma hominis (MH), Mycoplasma genitalium (MG), Chlamydia trachomatis (CT), herpes simplex virus-2 (HSV-2) and Neisseria gonorrhoeae (NG) were 8.7 %, 21.0 %, 8.2 %, 2.1 %, 3.6 %, 1.0 and 0 %, respectively. The STI detection rates of patients with LCS were higher than those of the non-LCS group (52.3 % vs. 39.3 %), although there was no statistically significant difference between the two groups (P = 0.07). All semen parameters were not significantly different between LCS with STIs and without STIs, except the semen volume in the MG-infected patients with LCS was significantly lower than that in the noninfected group. CONCLUSIONS: LCS was associated with a reduction in semen quality, but was not associated with STIs.

HPV infection and bacterial microbiota in the semen from healthy men.

BACKGROUND: Aberrant microbiota composition has been linked to disease development at numerous anatomical sites. Microbiota changes in reaction to viral infections, such as human papillomavirus (HPV), have been investigated almost exclusively in the female reproductive tract. However, HPV infection may also affect male health by reducing semen quality and fertility. The aim of this study was to investigate whether present HPV DNA is associated with detectable changes in semen bacterial microbiota composition and diversity. METHODS: This study relied on stored semen samples from 31 fertile healthy men who participated in the Finnish family HPV Study during the years 1998-2001. DNA was extracted from semen with PCR template preparation kit. HPV was genotyped using Luminex-based Multimetrix® assay. Microbiota was analyzed from the V3-V4 region of 16S rDNA gene following sequencing on an Illumina MiSeq platform. All statistical analyses were performed with Calypso software version 8.84. RESULTS: HPV DNA was detected in 19.4% (6/31) of the semen samples. HPV status in the semen did not impact the α-diversity estimations, as measured by Chao1 and Shannon indices, nor ß-diversity. Nevertheless, HPV-positive semen samples exhibited differences in the taxonomic composition of the bacterial microbiota including higher abundances of Moraxellaceae (p = 0.028), Streptococcus (p = 0.0058) and Peptostreptococcus (p = 0.012) compared to HPV-negative semen samples. CONCLUSION: HPV infection is associated with altered bacterial microbiota composition in semen, and this might have in impact to male health in general. As of present, it is unclear whether these changes result from HPV infection or whether altered bacterial microbiota increases susceptibility to HPV infection. More research is needed on viral-bacterial interactions in the male reproductive system.

Adjusted method of penis fixation during boar semi-automatic semen collection aiming to reduce bacterial contamination.

Semen collection has an essential role in the initial bacterial load in boar ejaculates and extended semen. The study aimed to explore the efficacy of an adjusted penis fixation in a semi-automatic collection system on reducing bacterial contamination of ejaculates in two-boar studs with different scenarios. Historically, stud A had low levels of bacterial load in raw semen, while stud B had a high level of contamination. A total of 56 mature boars had their semen collected using two methods of penis fixation: (a) Traditional: The penis was fixed directly with the artificial cervix and transferred to the adjustable clamp; (b) Adjusted: The fixation was performed with one gloved-hand, and after exteriorization, the penis was gripped using the artificial cervix with the other gloved-hand and transferred to the adjustable clamp. The bacterial load (p = .0045) and the occurrence of ejaculates >231 CFU/ml (p = .0101) were reduced in the Adjusted compared to the Traditional method. Bacterial load was reduced when using the Adjusted method in stud B (p = .0011), which showed a greater occurrence of critical factors for bacterial contamination (p ≤ .0034). The Adjusted method reduced the occurrence of ejaculates >231 CFU/ml when the preputial ostium was dirty (p = .016) and the duration of semen collection was >7 min (p = .022) compared to the Traditional method. In conclusion, the Adjusted penis fixation was efficient in reducing bacterial load of ejaculates, mainly in boar stud B, which had high contamination challenges.

Effects of air exposure and agitation on quality of stored boar semen samples.

The objective of this study was to investigate the effects of semen volume, air contact inside semen dose tubes, daily agitation of semen doses and extender type on semen quality, thermo-resistance and bacteria growth in extended boar semen doses preserved over 7 days of liquid storage. Ejaculates from 4 proven terminal cross-bred boars were collected using the gloved-hand technique for 4 weeks and used in the 3 × 2 × 2 factorial study. The effects of treatment (CON: 80 ml doses sealed at the top of the tube; 40HIGH: 40 ml doses sealed at top of tube, and 40LOW: 40 ml doses sealed at top of the liquid), agitation (agitated versus not agitated) and extender type (long-term versus short-term) were investigated on semen quality, thermo-resistance and bacteria growth in boar semen doses. The results of the study revealed that motility (p = .031) and viability (p = .041) in 40HIGH were lower than CON. pH (p < .001) was higher in 40HIGH compared with CON and 40LOW. Agitation did not impact motility (p = .581), progressive motility (p = .870), viability (p = .509) or morphology (p = .970), while long-term extender maintained higher motility (p = .002), progressive motility (p = .036), viability (p < .001) and normal acrosome (p < .001) than a short-term extender. VAP (p = .039) of 40HIGH was lower than CON in a thermo-resistance test. Neither treatment (p > .798, .766) nor agitation (p > .396, .476) impacted bacterial growth in this study. In conclusion, air contact negatively impacts boar semen pH and consequently sperm motility. Semen doses prepared with 80 or 40 ml volumes of extended boar semen with minimal air contact in the tubes yield more desirable semen quality and agitating boar semen doses daily does not have negative or positive effects on boar semen quality.

Total Aseptization of Boar Semen, to Increase the Biosecurity of Reproduction in Swine.

The aim of the study was to establish the complete microbiological profile of boar semen (Sus scrofa domesticus) and to choose the most effective antiseptic measures in order to control and optimize AI reproduction in pig farms. One hundred and one semen samples were collected and analyzed from several pig farms. The microbiological profile of ejaculates was determined by evaluating the degree of contamination of fresh semen and after dilution with specific extenders. The bacterial and fungal load of fresh boar semen recorded an average value of 82.41/0.149 × 103 CFU/mL, while after diluting the ejaculates the contamination value was 0.354/0.140 × 103 CFU/mL. Twenty-four germs (15 bacterial and 9 fungal species) were isolated, the most common being Candida parapsilosis/sake (92%) and Escherichia coli (81.2%). Modification of the sperm collection protocol (HPBC) reduced contamination in raw sperm by 49.85% in bacteria (significant (p < 0.00001) and by 9.67% in fungi (non-significant (p < 0.111491). The load in bacteria and filamentous fungi can be controllable, but not in levuras fungi. Some fluconazole-added extenders (12.5 mg%), ensure fungal aseptization, and even an increase in sperm progressivity (8.39%) for at least a 12 h shelf life after dilution. Validation of sperm aseptization was done by maintaining sow fecundity unchanged after AI (insignificant p > 0.05).

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

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.

Haemophilus seminalis sp. nov., isolated from human semen.

Two Haemophilus-like isolates with similar biochemical characteristics, designated strains SZY H1T and SZY H2, were isolated from human semen specimens. Cells were Gram-negative, non-motile, non-acid-fast, pleomorphic rods or coccobacilli. The major fatty acids (>10 %) were C16 : 0, C14 : 0, iso-C16 : 0 and/or C14 : 0 3-OH and C16 : 1 ω6c and/or C16 : 1 ω7c. The polar lipids were determined to be phosphatidylethanolamine, phosphatidylglycerol, an unidentified phospholipid, an unidentified aminophospholipid, two unidentified polar lipids and four unidentified aminolipids. The major polyamine was found to be cadaverine. The near-full-length (1462 nt) 16S rRNA gene sequences analysis showed the two isolates were nearly identical (>99.8 %), and closely matched Haemophilus haemolyticus ATCC 33390T with 98.9-99.1 % sequence similarities. Phylogenetic analysis based on 16S rRNA gene sequences and concatenation of 30 protein markers also revealed that the isolates clustered together with H. haemolyticus ATCC 33390T, and formed a distinct lineage well separated from the other members of the genus Haemophilus. Further, the average nucleotide identity values between the two isolates and their related species were below the established cut-off values for species delineation (95 %). Based on these findings, the two isolates are considered to represent a new species of the genus Haemophilus, for which name Haemophilus seminalis sp. nov. is proposed. The type strain is SZY H1T (=NBRC 113782T=CGMCC 1.17137T).

The microbiome of the infertile male.

PURPOSE OF REVIEW: Contrary to historic dogma, many tissues and organs in the human body contain a resident population of bacteria, fungi, and viruses collectively known as the microbiome. The microbiome plays a role in both homeostatic symbiosis and also pathogenic dysbiosis in a wide array of diseases. Our understanding of the relationship between the microbiome and male factor infertility is in its infancy but is slowly evolving. RECENT FINDINGS: Recent literature indicates that semen (and likely the testis) is not sterile and contains a distinct microbiome, and these changes in its composition are associated with alterations in semen quality and fertility status. Preliminary investigation indicates that manipulating the human microbiome may have implications in improving semen parameters and fertility. SUMMARY: In this review, we describe relationships between the microbiome and the genitourinary system, discuss the prior work on the relationship among bacteriospermia, leukocytospermia and male factor infertility, and summarize the current literature utilizing 16s rRNA-based next-generation sequencing on the seminal and testicular microbiome. We explore the specific microbial taxa implicated in various aspects of spermatic dysfunction and introduce preliminary evidence for therapeutic approaches to alter the microbiome and improve fertility status.

Antimicrobial resistance, genetic characterization, and molecular epidemiology of Ureaplasma species in males with infertility.

This study aimed to study the antimicrobial resistance, genetic characterization, and molecular epidemiology of Ureaplasma species in order to provide clinicians sufficient data to select optimal strategies of treatment for genitourinary tract infections of infertile male patients. Firstly, a total of 817 clinical semen specimens were detected for Ureaplasma species by molecular detection. Secondly, culture and identification of Ureaplasma species were achieved by using Mycoplasma ICS Test, and the antimicrobial susceptibility tests were determined by using broth microdilution assay. Then, the tetracycline resistance genetic determinants in Ureaplasma species were identified by PCR, and the fluoroquinolone and macrolide resistance genetic determinants were identified by DNA sequencing. Finally, the molecular epidemiology of Ureaplasma species was studied by both multilocus sequence typing (MLST) and expanded MLST (eMLST) schemes. Among the 817 semen specimens, 320 (39.17%) specimens were positive for Ureaplasma species. The percentages of resistance in 320 isolates against LEV, MXF, TET, and ERY were 47.5%, 39.38%, 19.69%, and 3.75%, respectively. The tet(M) and int-Tn genes were detected positive in all the tetracycline-resistant isolates. One macrolide-resistant UU isolate had a novel amino acid alteration (R66T) in L4 ribosomal protein and another UU isolate harbored a novel alteration (S109T) in L22. In fluoroquinolone-resistant isolates, S83L substitution in the ParC was predominant. In this area, ST22 and eST16 were the most prevalent ST and eST, respectively. One ST and 3 eSTs were newly identified in this study. This study has demonstrated that ERY can be first-line therapy for Ureaplasma species infections.

Composition of semen and foreskin mucosa aerobic microbiota and its impact on sperm parameters of captive collared peccaries (Pecari tajacu).

AIM: To evaluate the bacterial composition of collared peccary semen and foreskin mucosa, and to verify the sensitivity of isolates to antimicrobials used in semen conservation and to Aloe vera gel, which is an alternative external cryoprotectant. METHODS AND RESULTS: Nine foreskin mucosa and ejaculate samples from adult animals were used. Sperm characteristics and bacterial load were evaluated in fresh semen. The preputial mucosa and semen bacterial isolates were identified and tested against five concentrations of each antimicrobial (streptomycin-penicillin and gentamicin) and A. vera gel. Corynebacterium sp. and Staphylococcus sp. were isolated in greater numbers than others in both semen (64·10 and 20·51%, respectively) and the foreskin mucosa (60·60 and 24·25%, respectively), and ranged from 0·4 to 21 × 105 colony-forming units (CFU) per ml. The average load of Corynebacterium sp. was negatively correlated (P < 0·05) with the sperm membrane integrity (r = -0·73055) and curvilinear velocity (r = -0·69048). Streptomycin-penicillin and gentamicin inhibited most micro-organisms, and A. vera showed lower antimicrobial activity. CONCLUSION: Several Gram-positive bacteria are present in semen and foreskin mucosa of collared peccary, and the benefits of using primarily penicillin-streptomycin and gentamicin antimicrobials in the bacterial control of diluted semen of these animals are strongly indicated. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides insight into the reproductive microbiota of captive male-collared peccary. This work provides a theoretical basis to assist reproductive biotechnologies for ex situ conservation of the species.

Antibiotic resistance in microorganisms isolated in a bull semen stud.

Many microorganisms from various sources may be present in ejaculates of bulls. This study identified and isolated bacteria from bull sperm samples in a commercial stud and evaluated their resistance to antibiotics. The number of colony-forming units was determined in semen samples collected at distinct steps during freezing and thawing. The minimum inhibitory concentration and the minimum bactericidal concentration were determined for four antibiotics commonly used in commercial studs. A total of 135 microorganisms from 25 genera were isolated. After a sensitivity test, all evaluated microorganisms (n = 55) were resistant to penicillin and most of them were resistant to tylosin and lincomycin (n = 54). Resistance to all tested antibiotics was observed in 22% of all isolates, whereas only 3.9% of the isolates were inhibited by the tested antibiotics at the concentrations recommended by the international legislation. As the isolated microorganisms presented high resistance to frequently used antibiotics, sensitivity tests should be periodically conducted in commercial bull semen studs to prevent the use of contaminated semen in artificial insemination.

Characterization of semen collected by pharmacologically induced ejaculation from a stallion with seminal vesiculitis.

The present study compared the quality of sperm collected by artificial vagina or pharmacologically induced ejaculation from a 10-year-old thoroughbred stallion with seminal vesiculitis. The pharmacological protocol involved intravenous administration of detomidine (0.01 mg/kg) and oxytocin (20 IU) and successfully induced ejaculation in all attempts of semen collection. Sperm motility, plasma membrane and acrosome integrity (PMAI), reactive oxygen species (ROS) levels, polymorphonuclear neutrophil (PMN) percentage, and bacterial profiles of fresh and cooled semen (5°C for 24 hr) were evaluated. Semen obtained by the pharmacological method presented reduced seminal volume, decreased PMN percentage and superior sperm motility in cooled samples. Moreover, higher PMAI and lower ROS levels were observed in semen collected by the pharmacological method. Therefore, pharmacologically induced ejaculation is an alternative to obtain semen with minimal contamination and with sperm of superior quality and longevity from stallions with seminal vesiculitis.

[Bacterial culture of donor semen: Analysis of results from 2008 to 2018].

OBJECTIVE: To investigate the distribution of pathogenic bacteria in donor semen and the effect of bacterial infection on semen quality. METHODS: We performed bacterial culture on and counted the bacterial colonies (BC) in the semen samples collected from 4 897 sperm donors from 2008 to 2018 and divided them into groups A (BC <104 cfu/ml, n = 4 229), B (BC ≥104 cfu/ml, n = 150) and C (BC = 0 cfu/ml, n = 518). Using the biochemical reaction system of the French Biological Merry Emmanuel Company, we identified the bacterial species in group B, subjected all the semen samples to SCA computer assisted semen analysis, and compared the semen quality among different groups. RESULTS: In the 4 897 semen samples, hybrid bacterial contamination was found in 6 (0.12%) and non-hybrid bacteria in 4 379 (89.42%), including 150 (3.43%) in group B. In the semen samples with BC ≥104 cfu/ml, Gram-negative (G－) bacteria were observed in 104 (69.33%), mainly including Escherichia coli, followed by Proteusbacillus vulgaris and Enterobacteria, Gram-positive cocci (G+) in 39 (26.00%), G－ bacteria in 4 (2.67%) and Neisseria gonorrhoeae in 3 (2.00%). Compared with group C, groups A and B showed remarkably reduced total sperm count (P < 0.05) and percentage of progressively motile sperm (P < 0.05) but no statistically significant differences in the semen liquefaction time, semen PH value, total sperm motility or the percentage of morphologically normal sperm (P > 0.05). CONCLUSIONS: Bacterial culture of donor semen revealed a positive rate of 89.42% and varied the bacterial species, mainly including G－ bacteria. And the semen quality decreased with the increase of bacterial colonies.

Microbiological, molecular, and histopathological findings in goats experimentally infected with Actinobacillus seminis.

The objective of this study was to experimentally evaluate the pathogenicity of an Actinobacillus seminis isolate named SAAS01 in goats. Animals were challenged with 2 mL of a suspension containing 1,5 × 108 CFU/mL of A. seminis (SAAS01 isolate) through the intrapreputial, epididymis tail, and conjunctival routes. Epididymis and testicular fragments were submitted to histopathological exam, and semen samples underwent microbiological and molecular diagnoses. Clinically, a unilateral increase in firm consistency was observed in the epididymis and testicles of two animals inoculated in epididymis tail and in one animal inoculated through conjunctival sac; this firmness continued until the day of euthanasia. Two goats inoculated through epididymis tail and conjunctival sac routes presented histopathological findings with macroscopically and microscopically significant changes. A. seminis was isolated from semen samples collected from goats inoculated through the epididymis tail and conjunctival sac routes. A. seminis DNA was amplified from six semen samples of three goats inoculated through the epididymis tail, two in conjunctival sac and one through intrapreputial route. The experimental infection model using goats confirmed the pathogenicity of the A. seminis isolate, demonstrating the predilection of the agent for the epididymis, with clinical signs, histopathological lesions, bacterial isolation, and a positive molecular diagnosis.

Treponema pallidum subsp. pallidum DNA and RNA in Semen of a Syphilis Patient Without Genital or Anal Lesions.

Treponema pallidum subsp. pallidum DNA and RNA were detected in a semen specimen of a syphilis patient with no genital or anal sores and no clinically evident orchitis. No nucleic acids were found in a urine sample of the same patient collected immediately before the semen sample. Exposure to the syphilis agent through semen could account for transmission episodes in the absence of direct contact with a syphilitic sore.