Undiagnosed RASopathies in infertile men.

RASopathies are syndromes caused by congenital defects in the Ras/mitogen-activated protein kinase (MAPK) pathway genes, with a population prevalence of 1 in 1,000. Patients are typically identified in childhood based on diverse characteristic features, including cryptorchidism (CR) in >50% of affected men. As CR predisposes to spermatogenic failure (SPGF; total sperm count per ejaculate 0-39 million), we hypothesized that men seeking infertility management include cases with undiagnosed RASopathies. Likely pathogenic or pathogenic (LP/P) variants in 22 RASopathy-linked genes were screened in 521 idiopathic SPGF patients (including 155 CR cases) and 323 normozoospermic controls using exome sequencing. All 844 men were recruited to the ESTonian ANDrology (ESTAND) cohort and underwent identical andrological phenotyping. RASopathy-specific variant interpretation guidelines were used for pathogenicity assessment. LP/P variants were identified in PTPN11 (two), SOS1 (three), SOS2 (one), LZTR1 (one), SPRED1 (one), NF1 (one), and MAP2K1 (one). The findings affected six of 155 cases with CR and SPGF, three of 366 men with SPGF only, and one (of 323) normozoospermic subfertile man. The subgroup "CR and SPGF" had over 13-fold enrichment of findings compared to controls (3.9% vs. 0.3%; Fisher's exact test, p = 5.5 × 10-3). All ESTAND subjects with LP/P variants in the Ras/MAPK pathway genes presented congenital genitourinary anomalies, skeletal and joint conditions, and other RASopathy-linked health concerns. Rare forms of malignancies (schwannomatosis and pancreatic and testicular cancer) were reported on four occasions. The Genetics of Male Infertility Initiative (GEMINI) cohort (1,416 SPGF cases and 317 fertile men) was used to validate the outcome. LP/P variants in PTPN11 (three), LZTR1 (three), and MRAS (one) were identified in six SPGF cases (including 4/31 GEMINI cases with CR) and one normozoospermic man. Undiagnosed RASopathies were detected in total for 17 ESTAND and GEMINI subjects, 15 SPGF patients (10 with CR), and two fertile men. Affected RASopathy genes showed high expression in spermatogenic and testicular somatic cells. In conclusion, congenital defects in the Ras/MAPK pathway genes represent a new congenital etiology of syndromic male infertility. Undiagnosed RASopathies were especially enriched among patients with a history of cryptorchidism. Given the relationship between RASopathies and other conditions, infertile men found to have this molecular diagnosis should be evaluated for known RASopathy-linked health concerns, including specific rare malignancies.

Toward clinical exomes in diagnostics and management of male infertility.

Infertility, affecting ∼10% of men, is predominantly caused by primary spermatogenic failure (SPGF). We screened likely pathogenic and pathogenic (LP/P) variants in 638 candidate genes for male infertility in 521 individuals presenting idiopathic SPGF and 323 normozoospermic men in the ESTAND cohort. Molecular diagnosis was reached for 64 men with SPGF (12%), with findings in 39 genes (6%). The yield did not differ significantly between the subgroups with azoospermia (20/185, 11%), oligozoospermia (18/181, 10%), and primary cryptorchidism with SPGF (26/155, 17%). Notably, 19 of 64 LP/P variants (30%) identified in 28 subjects represented recurrent findings in this study and/or with other male infertility cohorts. NR5A1 was the most frequently affected gene, with seven LP/P variants in six SPGF-affected men and two normozoospermic men. The link to SPGF was validated for recently proposed candidate genes ACTRT1, ASZ1, GLUD2, GREB1L, LEO1, RBM5, ROS1, and TGIF2LY. Heterozygous truncating variants in BNC1, reported in female infertility, emerged as plausible causes of severe oligozoospermia. Data suggested that several infertile men may present congenital conditions with less pronounced or pleiotropic phenotypes affecting the development and function of the reproductive system. Genes regulating the hypothalamic-pituitary-gonadal axis were affected in >30% of subjects with LP/P variants. Six individuals had more than one LP/P variant, including five with two findings from the gene panel. A 4-fold increased prevalence of cancer was observed in men with genetic infertility compared to the general male population (8% vs. 2%; p = 4.4 × 10-3). Expanding genetic testing in andrology will contribute to the multidisciplinary management of SPGF.

Lactobacillus crispatus-dominated vaginal microbiome and Acinetobacter-dominated seminal microbiome support beneficial ART outcome.

INTRODUCTION: Despite the considerable progress made in assisted reproductive technologies (ART), the implantation rate of transferred embryos remains low and in many cases, the reasons for failure remain unclear. We aimed to determine the potential impact of female and male partners' reproductive tract microbiome composition on ART outcome. MATERIAL AND METHODS: The ART couples (n = 97) and healthy couples (n = 12) were recruited into the study. The smaller healthy group underwent a careful selection according to their reproductive and general health criteria. Both vaginal and semen samples were subjected to 16S rDNA sequencing to reveal the bacterial diversity and identify distinct microbial community types. Ethics statement The study was approved by the Ethics Review Committee on Human Research of Tartu University, Tartu, Estonia (protocol no. 193/T-16) on 31 May 2010. Participation in the research was voluntary. Written informed consent was obtained from all study participants. RESULTS: The men with Acinetobacter-associated community who had children in the past, had the highest ART success rate (P < 0.05). The women with bacterial vaginosis vaginal microbiome community and with L. iners-predominant and L. gasseri-predominant microbiome had a lower ART success rate than women with the L. crispatus-predominant or the mixed lactic-acid-bacteria-predominant type (P < 0.05). The 15 couples where both partners had beneficial microbiome types had a superior ART success rate of 53%, when compared with the rest of the couples (25%; P = 0.023). CONCLUSIONS: Microbiome disturbances in the genital tract of both partners tend to be associated with couple's infertility as well as lower ART success levels and may thus need attention before the ART procedure. The incorporation of genitourinary microbial screening as a part of the diagnostic evaluation process may become routine for ART patients if our results are confirmed by other studies.

Microdeletions and microduplications linked to severe congenital disorders in infertile men.

Data on the clinical validity of DNA copy number variants (CNVs) in spermatogenic failure (SPGF) is limited. This study analyzed the genome-wide CNV profile in 215 men with idiopathic SPGF and 62 normozoospermic fertile men, recruited at the Andrology Clinic, Tartu University Hospital, Estonia. A two-fold higher representation of > 1 Mb CNVs was observed in men with SPGF (13%, n = 28) compared to controls (6.5%, n = 4). Seven patients with SPGF were identified as carriers of microdeletions (1q21.1; 2.4 Mb) or microduplications (3p26.3, 1.1 Mb; 7p22.3-p22.2, 1.56 Mb; 10q11.22, 1.42 Mb, three cases; Xp22.33; 2.3 Mb) linked to severe congenital conditions. Large autosomal CNV carriers had oligozoospermia, reduced or low-normal bitesticular volume (22-28 ml). The 7p22.3-p22.2 microduplication carrier presented mild intellectual disability, neuropsychiatric problems, and short stature. The Xp22.33 duplication at the PAR1/non-PAR boundary, previously linked to uterine agenesis, was detected in a patient with non-obstructive azoospermia. A novel recurrent intragenic deletion in testis-specific LRRC69 was significantly overrepresented in patients with SPGF compared to the general population (3.3% vs. 0.85%; χ2 test, OR = 3.9 [95% CI 1.8-8.4], P = 0.0001). Assessment of clinically valid CNVs in patients with SPGF will improve their management and counselling for general and reproductive health, including risk of miscarriage and congenital disorders in future offspring.

A common 1.6 mb Y-chromosomal inversion predisposes to subsequent deletions and severe spermatogenic failure in humans.

Male infertility is a prevalent condition, affecting 5-10% of men. So far, few genetic factors have been described as contributors to spermatogenic failure. Here, we report the first re-sequencing study of the Y-chromosomal Azoospermia Factor c (AZFc) region, combined with gene dosage analysis of the multicopy DAZ, BPY2, and CDYgenes and Y-haplogroup determination. In analysing 2324 Estonian men, we uncovered a novel structural variant as a high-penetrance risk factor for male infertility. The Y lineage R1a1-M458, reported at >20% frequency in several European populations, carries a fixed ~1.6 Mb r2/r3 inversion, destabilizing the AZFc region and predisposing to large recurrent microdeletions. Such complex rearrangements were significantly enriched among severe oligozoospermia cases. The carrier vs non-carrier risk for spermatogenic failure was increased 8.6-fold (p=6.0×10-4). This finding contributes to improved molecular diagnostics and clinical management of infertility. Carrier identification at young age will facilitate timely counselling and reproductive decision-making.

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.

Seminal microbiome in men with and without prostatitis.

OBJECTIVES: To profile the seminal microbiome applying next generation sequencing. METHODS: Semen samples of 67 men were involved in the study (21 men with and 46 men without prostatitis). Seminal microbiomes were profiled applying the method that uses combinatorial sequence tags attached to polymerase chain reaction primers that amplify the ribosomal ribonucleic acid V6 region. Amplified polymerase chain reaction products were sequenced using an Illumina paired-end protocol on HiSeq2000 platform. RESULTS: The most abundant phylum in semen was Firmicutes, comprising nearly half of the sequences found (median 41.7%, quartiles 28.5-47.2%) followed by Bacteroidetes, Proteobacteria and Actinobacteria. The counts of lactobacilli were higher in healthy men than prostatitis patients (27% [20.2-34.6%] vs 20.2% [4.9-25.0%]; P = 0.05), especially for Lactobacillus iners. Proteobacteria comprised higher proportions in prostatitis patients than healthy men. The species richness was higher in prostatitis patients than healthy men (inverted Simpson index 13.5 ± 5.8 vs 10.3 ± 4.0). CONCLUSIONS: The semen of chronic prostatitis patients contains fewer health-supporting lactobacilli, and has higher species diversity than that of healthy men. Firmicutes (especially lactobacilli), Bacteroidetes, Proteobacteria and Actinobacteria comprise the highest proportion of seminal microbiome.

Complementary seminovaginal microbiome in couples.

The genital tract microbiome is tightly associated with reproductive health. Although many research studies have been performed on the vaginal microbiome, current knowledge of the male microbiome is scarce, and parallel studies examining couples are extremely rare. In this work, we aimed to compare seminal and vaginal microbiomes in couples and to assess the influence of sexual intercourse on vaginal microbiome. The study included 23 couples. Microbiomes of semen and vaginal fluid (pre- and post-intercourse) were profiled using Illumina HiSeq2000 sequencing of the V6 region of 16S rRNA gene. Seminal communities were significantly more diverse, but with lower total bacterial concentrations than those of the vagina. Gardnerella vaginalis was predominant in half of the women whose partners had significant leukocytospermia, but only in one of 17 women who had a partner without leukocytospermia. There was significant decrease in the relative abundance of Lactobacillus crispatus after intercourse, and high concordance between semen and vaginal samples. Our data support the hypothesis that semen and vaginal microbiomes are in association, inasmuch as the predominance of G. vaginalis in female partners was significantly related to inflammation in male genital tracts.

Semen quality in middle-aged males: associations with prostate-specific antigen and age-related prostate conditions.

OBJECTIVE: To compare markers of semen quality and related reproductive indicators in middle-aged males in relation to serum prostate-specific antigen (PSA) levels. METHODS: A total of 384 voluntary middle-aged men who underwent screening for prostate health were recruited. Reproductive function, semen quality, hormonal parameters, prostate-related pathologies, and PSA levels were measured. RESULTS: Total sperm count and sperm concentration were decreased in men with elevated serum PSA compared with age-matched controls. Serum FSH levels differed significantly in subjects with non-malignant, pre-malignant, and malignant prostate conditions. PSA levels in men with three normal semen variables (semen volume ≥ 1.5 mL, sperm density ≥ 15 × 10(6)/mL, and A + B motility ≥ 40%) were significantly lower compared with subjects with altered parameters (1.51 ng/ml vs. 2.47 ng/ml, respectively, p = 0.002). PSA showed a negative correlation with semen volume and total sperm count, and a positive correlation with semen interleukin-6. CONCLUSIONS: Our data demonstrate that serum PSA levels are related to impaired reproductive parameters in middle-aged subjects. While there is no consensus about the potential link between male factor infertility, impaired reproductive indicators, and prostate pathologies, this topic needs additional research focusing on the interactions between male aging, reproductive function, and the prostate.

Decline of seminal parameters in middle-aged males is associated with lower urinary tract symptoms, prostate enlargement and bladder outlet obstruction.

PURPOSE: We aimed to compare the associations between semen quality, associated reproductive indicators and the main prostate-related parameters in middle-aged men. MATERIALS AND METHODS: This is a prospective study on 422 middle-aged men who underwent the screening for prostate health. Their reproductive function, semen quality and prostate-related pathologies were investigated. RESULTS: Significant associations between semen quality and prostate-related parameters could be seen. Total sperm count and sperm density decreased along with the increase of the I-PSS score and total prostate volume. Also, the related lower urinary tract characteristics showed a negative correlation with main semen parameters for all investigated subjects. No significant differences in age, testicular size, and hormonal parameters were found between the subjects with or without lower urinary tract symptoms and prostate enlargement. CONCLUSIONS: Our study suggests that altered seminal parameters in middle-aged men are associated with LUTS, prostate enlargement and/or bladder outlet obstruction. Although the assessments of prostate and lower urinary tract symptoms may not replace the semen parameters evaluating the male reproductive status, there is a need for further and more detailed investigations about the pathways behind these associations as well as possible related conditions.

Decline of seminal parameters in middle-aged males is associated with lower urinary tract symptoms, prostate enlargement and bladder outlet obstruction

Purpose We aimed to compare the associations between semen quality, associated reproductive indicators and the main prostate-related parameters in middle-aged men. Materials and Methods: This is a prospective study on 422 middle-aged men who underwent the screening for prostate health. Their reproductive function, semen quality and prostate-related pathologies were investigated. Results Significant associations between semen quality and prostate-related parameters could be seen. Total sperm count and sperm density decreased along with the increase of the I-PSS score and total prostate volume. Also, the related lower urinary tract characteristics showed a negative correlation with main semen parameters for all investigated subjects. No significant differences in age, testicular size, and hormonal parameters were found between the subjects with or without lower urinary tract symptoms and prostate enlargement. Conclusions Our study suggests that altered seminal parameters in middle-aged men are associated with LUTS, prostate enlargement and/or bladder outlet obstruction. Although the assessments of prostate and lower urinary tract symptoms may not replace the semen parameters evaluating the male reproductive status, there is a need for further and more detailed investigations about the pathways behind these associations as well as possible related conditions. .

Semen quality and associated reproductive indicators in middle-aged males: the role of non-malignant prostate conditions and genital tract inflammation.

PURPOSE: To compare the associations between non-malignant prostate conditions, genital tract inflammation, and reproductive function in middle-aged men. METHODS: Three-hundred and eighty-two voluntary male subjects who underwent the screening for prostate health were recruited for the study. Semen quality and associated reproductive indicators, seminal inflammation, and prostate-related pathologies were evaluated. RESULTS: Sperm motility and prostate-related parameters were significantly impaired in patients with chronic prostatitis syndromes and lower urinary tract symptoms in comparison with controls. Elevated seminal markers of inflammation were in positive association with body mass index, prostate-specific antigen, and estradiol level in serum while in negative association with semen volume, total sperm count, and sperm motility. According to WHO reference limits, speculative cutoff values for WBC and IL-6 in semen to detect reduced sperm parameters were 0.342 M/mL and 56.8 ng/L, respectively. CONCLUSIONS: According to our data, one of the possible pathways for impaired reproductive quality in male subjects >45 years could be related to infection and inflammation in the genital tract with subsequent (partial) obstruction and damage of prostate and other male accessory glands.

Balanced reciprocal translocation t(5;13)(q33;q12) and 9q31.1 microduplication in a man suffering from infertility and pollinosis.

We describe the first case of two chromosomal abnormalities, balanced reciprocal translocation t(5;13)(q33;q12.1) and a microduplication in the region 9q31.1, in a man suffering from infertility and pollinosis. In the region 13q12.1 is located the TUBA3C (tubulin, alpha 3c) gene, which plays an important dynamic role in the motility of flagella. This case might support the opinion that haploinsufficiency of the TUBA3C gene could be the cause of sperm immotility and abnormal sperm morphology, resulting in infertility in the patient. Single-nucleotide polymorphism (SNP) array analysis revealed a novel 9q31.1 microduplication inherited from both parents, which contributes to the genomic instability.

Influence of sexual intercourse on genital tract microbiota in infertile couples.

Several studies have suggested the association of disturbed genital tract microbiota with infertility. Our aim was to clarify the influence of sexual intercourse on partner's genital tract microbiota in infertile couples. Seventeen couples were studied, and in 5 men inflammatory prostatitis (IP) was diagnosed. Semen samples were collected during menstruation of the female counterpart, two self-collected vaginal samples were taken 3-5 days later - before intercourse and 8-12 h after intercourse. Ureaplasma parvum was found in 59% of women, its prevalence was higher in women whose partner had IP, as well as in half of their male partners. Sexual intercourse caused significant shifts in vaginal microbiota - increase of Nugent score and shifts in cultured microbiota (emergence and disappearance of several species). These changes were less expressed in the presence of normal vaginal microbiota but more prominent in the partners of IP men. These changes may interfere with fertilization.

Seminal interleukin-6 and serum prostate-specific antigen as possible predictive biomarkers in asymptomatic inflammatory prostatitis.

OBJECTIVES: To determine the possible predictive values of seminal interleukin-6 (IL-6) and serum prostate-specific antigen (PSA), as well as their combined values, in differentiating between subjects with or without asymptomatic inflammatory prostatitis. METHODS: The study group consisted of 490 men (mean age 18.9 ± 1.8 years, range 16-25). Cytologic examination of all ejaculates (using Bryan-Leishman-stained slides) and clinical examination for possible pathologic findings in the genital region were performed. The subjects with any clinical symptoms of inflammation were excluded. The levels of PSA in the blood serum and IL-6 in the seminal plasma were also determined. The IL-6 and PSA levels for different leukocytospermia status were statistically compared, and receiver operating characteristic curves were designed to determine the sensitivity versus specificity and the positive and negative predictive values of IL-6 and PSA levels against different thresholds of leukocytospermia (0.2, 0.5, and >1.0 × 10(6) leukocytes/mL). RESULTS: The levels of both IL-6 in the seminal plasma and PSA in the blood serum were significantly greater in National Institutes of Health prostatitis IV than in the controls. The receiver operating characteristic curves for seminal IL-6 and serum PSA showed high negative prognostic values for all 3 leukocytospermic subgroups, and positive prognostic values were seen only with IL-6 in the lower leukocytospermic range. CONCLUSIONS: Both seminal IL-6 and serum PSA are excellent negative predictive markers for asymptomatic inflammatory prostatitis in young men, although positive predictive values of these biomarkers remain less indicative in this age group.

Increased Prevalance of the -211 T allele of follicle stimulating hormone (FSH) beta subunit promoter polymorphism and lower serum FSH in infertile men.

CONTEXT: The human FSHB promoter polymorphism (rs10835638; -211 G/T) has been associated with serum FSH in a cohort of young Estonian men. The minor allele carriers had reduced serum FSH (15.7% in GT heterozygotes; 40% in TT homozygotes) compared with GG homozygotes. OBJECTIVE: Because FSH is essential for normal spermatogenesis and fertility, we speculated that abnormalities in FSH action could contribute to male infertility. We sought to study whether genetically inherited constitutively reduced FSH levels may affect male reproduction and replicate the association between rs10835638 and serum FSH among infertile male patients. DESIGN: Genotyping of rs10835638 in a cohort of infertile men (n = 1029; Andrology Center of the Tartu University Clinics, Estonia), including idiopathic infertility cases (IIFC; n = 750). PATIENTS: Patients included male partners (sperm concentration <20 x 10(6)/ml) of infertile couples failing to conceive a child for 12 months or longer. RESULTS: A significant excess of TT homozygotes (1.1 vs. 2.4%) as well as GT heterozygotes (22.4 vs. 25.1%) was detected among infertile men compared with the young male cohort (chi(2) test, P < 0.05). The T allele of rs10835638 was associated with reduced serum FSH (analysis of covariance; full cohort: P = 1.20 x 10(-6), F = 13.8; IIFC: P = 7.70 x 10(-7), F = 14.3) as well as with low FSH to LH ratio (full cohort: P = 1.52 x 10(-11), F = 25.6; IIFC: P = 3.25 x 10(-9), F = 20.4). The median serum FSH levels differed between the GG and TT carriers by 48.5%. All IIFC with TT genotype exhibited low (<1.8) FSH to LH ratio. CONCLUSIONS: In perspective, this genetic marker may have clinical significance in molecular diagnostics of male reproductive success and a potential to identify positive responders to FSH treatment.

Prevalence of asymptomatic inflammatory (National Institutes of Health Category IV) prostatitis in young men according to semen analysis.

OBJECTIVES: To determine the prevalence of asymptomatic inflammatory (National Institutes of Health category IV) prostatitis in a cross-sectional community sample of young men. METHODS: The study group consisted of 565 men aged 18.9 +/- 1.8 years (mean +/- standard deviation). Cytologic examination of all ejaculates (using Bryan-Leishman stained slides), as well as analysis for basic semen parameters (volume, concentration, and motility) and clinical examination for possible pathologies in the genital region, were performed. Subjects with any clinical symptoms of inflammation were excluded. Levels of prostate-specific antigen in blood serum and interleukin-6 in seminal plasma were determined as well. RESULTS: The prevalence of asymptomatic inflammatory prostatitis (greater than 1 x 10(6) white blood cells [WBC]/mL in semen, according to World Health Organization guidelines) was 6.0%, but when we used a lower threshold suggested by our previous studies (greater than 0.2 x 10(6) WBC/mL), the prevalence was 19.0%. In this study the analysis did not show any significant effect of leukocytospermia on sperm quality, except higher sperm concentration in subjects with moderate leukocytospermia (0.2-1 x 10(6) WBC/mL). We did not detect any seasonal differences in the prevalence of asymptomatic inflammatory prostatitis. Interleukin-6 and prostate-specific antigen levels were significantly higher in leukocytospermic subjects than in those without leukocytospermia. CONCLUSIONS: Asymptomatic inflammatory prostatitis has a notable prevalence among healthy young men, suggesting the need for further studies to investigate pathogenetic mechanisms of the disease. Lack of negative effect of leukocytospermia on basic semen parameters may indicate higher functional reserve of accessory sex glands in this age group.

Coryneform bacteria in semen of chronic prostatitis patients.

Our aim was to compare the presence and species composition of coryneform bacteria in chronic prostatitis patients and controls. Semen of 50 men with inflammatory prostatitis and 59 controls (without pelvic pain/discomfort complaints and leukocytospermia) was investigated. First-catch urine was additionally investigated in 36 men (30 with and 6 without prostatitis). Coryneform bacteria were found in semen of 76% men with inflammatory prostatitis and 83% controls. More than half of the isolates were identified as Corynebacterium seminale. Prostatitis patients with severe leukocytospermia (>1 million white blood cells per ml) harboured significantly more Corynebacterium group G (33% vs. 2%, p = 0.0003) and Arthrobacter sp. (17% vs. 2%, p = 0.03) in comparison with controls. Nine species of coryneforms with high concentration (>or=10,000 CFU per ml) were found in prostatitis patients as against only four species in controls. Half of the men harboured corynebacteria in semen as well as in urine, 22% of men in semen only, and 3% in urine only. The total concentration of coryneforms was greater in semen than in urine (median 5000 vs. 100 CFU per ml, p = 0.053). We suggest that although coryneforms are generally considered as saprophytes, they are not uniform and some species (Corynebacterium group G and Arthrobacter sp.) may be associated with inflammatory prostatitis.

Seminal microflora in asymptomatic inflammatory (NIH IV category) prostatitis.

OBJECTIVES: To prove the need for the quantitative full-microflora semen analysis for determining the role of microorganisms in the etiology of asymptomatic inflammatory prostatitis, and to correlate the seminal white blood cell (WBC) counts with interleukin 6 (IL-6) levels. METHODS: Thirty-seven men with asymptomatic inflammatory (National Institutes of Health [NIH] IV category) prostatitis and 32 controls were investigated by using routine semen analysis, IL-6 levels of seminal plasma, and quantitative microbiological analysis of semen. RESULTS: The IL-6 concentration in seminal plasma was significantly higher in NIH IV category prostatitis patients than in the controls, and was in good correlation with the WBC count in semen (r=0.74, p<0.001). In most of the specimens, the counts of anaerobic microorganisms were equal to or outnumbered the aerobic ones. One to eight different microorganisms could be found in any particular semen sample, and the total concentration of microorganisms ranged from 2.0 to 7.5log(10)CFU/ml. Both parameters were significantly higher in NIH IV category prostatitis patients than in controls (median: 4.8 vs. 3.9log(10)CFU for total concentration, p<0.001; median: 5 vs. 3 for number of different species, p=0.004). CONCLUSIONS: Unlike the controls the NIH IV category prostatitis patients harbour abundant polymicrobial microbiocenosis in their semen, containing anaerobic, microaerophilic and aerobic bacteria. Detection of IL-6 in seminal plasma serves as an additional tool for diagnosing NIH IV category prostatitis.