OBJECTIVETo improve the sample collection methods and bacteriologic localization patterns in male genital tract infection, and to investigate the influence of specimen collection and pathogen isolation on the diagnosis and treatment of prostatitis.

METHODSWe collected the samples of the initial urinary stream, the third portion of the urinary stream, expressed prostatic secretion (ESP), and semen from 200 adult males with chronic prostatitis-like symptoms, inoculated them quantitatively in culture media for isolation of microorganisms, and evaluated their laboratory diagnostic significance according to the count of colonies and distribution of the isolates.

RESULTSA total of 468 strains of microorganisms were isolated from the samples, including 414 strains of bacteria spp (88.5%), 12 strains of fungi spp (2.6%), 40 strains of mycoplasma spp (8.5%), and 2 strains of chlamydia spp (0.4%). Pathogens were isolated from the ESP in 66 cases (33.0%), from the semen in 34 cases (17.0%), and from both the ESP and semen in 100 cases (50.0%). Only 1 species of pathogen was found in the ESP samples of 36 cases (18.0%), in the semen samples of 20 cases (10%), and in both the ESP and semen samples of 39 cases (19.5%); 2 species in the ESP samples of 30 cases (15.0%), in the semen samples of 14 cases (7.0%), and in both the ESP and semen samples of 60 cases (30.0%); and 3 species in both the ESP and semen samples of 1 case (0.5%).

CONCLUSIONMultiple microbial infection (MMI), multi-organ infection (MOI) and drug-resistance strains infection are common in patients with prostatitis-like symptoms, frequently leading to missed diagnosis and misdiagnosis in clinic and laboratory, and affecting the effect of antimicrobial therapy. MMI and MOI can be diagnosed and differentially diagnosed with the improved sample collection methods and bacteriologic localization patterns.

Adult ; Bacterial Load ; Chronic Disease ; Genital Diseases, Male ; diagnosis ; microbiology ; Humans ; Male ; Prostatitis ; diagnosis ; microbiology ; Reproductive Tract Infections ; diagnosis ; microbiology ; Semen ; microbiology ; Specimen Handling ; methods ; Urethra ; microbiology

Objective: To analyze the relationship of Mycoplasma genitalium (MG) infection with routine semen parameters and sperm DNA integrity in male infertility patients. METHODS: Totally, 114 semen samples, 34 MG-positive and 80 MG-negative, were collected from male infertility patients and subjected to routine semen analysis with the computer-assisted sperm analysis system, Papanicolaou staining for observation of sperm morphology, and sperm chromatin diffusion (SCD) test for detection of sperm DNA integrity. Semen parameters and DNA integrity were compared between the MG-positive and MG-negative groups with SPSS 21.0 statistical software and the relationship between the semen parameters and DNA integrity analyzed by Pearson correlation analysis. RESULTS: The MG-positive samples, compared with the MG-negative ones, showed significantly decreased semen volume (［2.87 ± 0.37］ vs ［3.86 ± 0.43］ ml, P < 0.01), sperm concentration (［29.05 ± 6.17］ vs ［32.56 ± 5.97］ ×10⁶/ml, P < 0.01), and percentages of progressively motile sperm (PMS) (［15.86 ± 2.79］% vs ［23.65 ± 3.47］%, P < 0.01) and morphologically normal sperm (MNS) (［6.35 ± 2.06］% vs ［7.14 ± 1.89］%, P < 0.05), increased proportions of non-halo sperm (［15.02 ± 3.52］% vs ［9.72 ± 2.94］%, P <0.01) and small-halo sperm (［16.37 ± 5.26］% vs ［11.07 ± 1.65］%, P < 0.01) and sperm DNA fragmentation index (DFI) (［31.39 ± 3.16］% vs ［20.79 ± 3.59］%, P < 0.01), and reduced proportion of large-halo sperm (［54.75 ± 8.74］% vs ［64.15 ± 9.76］%, P < 0.01). DFI was negatively correlated with the percentages of PMS (r = －0.516, P < 0.05) and MNS (r = －0.429, P < 0.05) in the MG-positive group, but not correlated with any of the routine semen parameters in the MG-negative patients (P > 0.05). CONCLUSIONS MG infection may be an important factor affecting sperm quality in male infertility patients. Active prevention and treatment of MG infection can help prevent male infertility.
DNA Fragmentation ; Humans ; Infertility, Male/microbiology* ; Male ; Mycoplasma Infections/complications* ; Mycoplasma genitalium ; Semen ; Semen Analysis ; Sperm Count ; Sperm Motility ; Spermatozoa

Objective: To investigate the value of real-time RNA simultaneous amplification and testing (SAT) in the detection of Ureaplasma urealyticum (UU) in the semen of infertile males and its clinical significance. METHODS: We collected semen samples from 542 infertility patients and 120 normal fertile men as controls in the Andrology Clinic of Nanjing General Hospital from March to September 2015. We detected UU infection in the samples using the culture method and SAT technology, respectively. RESULTS: All the UU positive cases (except 4 false positive cases) detected by the culture method were also shown to be positive in SAT. The UU detection rate of SAT was significantly higher than that of the culture method both in the infertility patients (54.1 vs 19.7%, P<0.05) and in the normal controls (42.5 vs 12.5%, P<0.05). CONCLUSIONS SAT is a rapid and accurate method for detecting UU infection in semen samples, with a higher sensitivity and accuracy than the culture method, and it can also be used to evaluate the therapeutic effects. However, the culture method has its own advantages, such as low requirement of technical equipment, easy operation, and possibility of drug sensitivity test at the same time. Therefore, SAT and the culture method can be used alternatively according to the clinical need.
Andrology ; Humans ; Infertility, Male ; microbiology ; Male ; Nucleic Acid Amplification Techniques ; RNA, Bacterial ; analysis ; Semen ; chemistry ; microbiology ; Semen Analysis ; Ureaplasma Infections ; diagnosis ; Ureaplasma urealyticum ; genetics ; isolation & purification

ObjectiveTo explore Mycoplasma genitalium (MG) infection in the urogenital tract of infertile men and its influence on semen quality.

METHODSSemen samples were collected from 352 infertile males in the Center of Reproductive Medicine of Nanjing General Hospital from March to July 2015. MG infection was detected by real-time fluorescence simultaneous amplification and testing and semen analyses were conducted according to the WHO Laboratory Manual for the Examination and Processing of Human Semen (5th Ed) on the semen pH value, semen volume, total sperm count, sperm concentration, total sperm motility, percentages of progressively motile sperm (PMS) and immotile sperm (IMS), and sperm DNA fragmentation index (DFI). The data obtained were subjected to statistical analysis by t-test and non-parametric test (Wilcoxon test).

RESULTSMG infection was found in 3.4% (12/352) of the infertile patients. Compared with the MG-positive cases, the MG-negative ones showed a significantly higher semen volume (［2.85 ± 0.14］ vs ［3.84 ± 0.12］ ml, P = 0.008) and percentage of PMS (［15.86±1.72］ vs ［60.95 ± 5.63］ %, P = 0.032) but a lower DFI (［30.73 ±2.24］ vs ［20.71 ± 1.55］%, P = 0.014). However, no statistically significant differences were observed between the two groups in the semen pH value (7.38 ±0.02 vs 7.39 ± 0.01, P = 0.774), sperm concentration (［52.96 ± 15.78］ vs ［60.05 ± 4.29］×10⁶/ml, P = 0.683), sperm count (［154.15 ± 46.37］ vs ［221.56 ± 15.43］×106, P = 0.236), total sperm motility (［29.04 ± 3.11］ vs ［33.52 ± 1.51］ %, P = 0.626), or percentage of IMS (［23.57 ± 0.99］ vs ［62.34 ± 1.69］ %, P = 0.691).

CONCLUSIONSUrogenital MG infection is common in infertile males and potentially affects the semen quality, especially sperm vitality of the patient.

DNA Fragmentation ; Humans ; Infertility, Male ; microbiology ; physiopathology ; Male ; Male Urogenital Diseases ; microbiology ; Mycoplasma Infections ; complications ; Mycoplasma genitalium ; Semen ; Semen Analysis ; Sperm Count ; Sperm Motility ; Spermatozoa ; physiology

OBJECTIVES: To investigate the specific microbial signatures in vaginal fluid. METHODS: Vaginal fluid （16 samples）, saliva （16 samples）, feces （16 samples）, semen （8 samples）, peripheral blood （8 samples）, urine （5 samples）, and nasal secretion （4 samples） were collected respectively. The 16S rRNA genes of Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus jensenii, Lactobacillus iners, and Atopobium vaginae were amplified. PCR production was detected via a 3130xl Genetic Analyzer. RESULTS: The detected number of Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus jensenii, Lactobacillus iners, and Atopobium vaginae were 15, 5, 8, 14, and 3 in all vaginal fluid samples, respectively. Lactobacillus crispatus and Lactobacillus jensenii existed specifically in vaginal fluid. CONCLUSIONS There is a potential application value to detect Lactobacillus crispatus and Lactobacillus jensenii for the identification of vaginal fluid.
Actinobacteria/classification* ; Blood/microbiology* ; Body Fluids/microbiology* ; Feces/microbiology* ; Female ; Genes, Bacterial ; Humans ; Lactobacillus/classification* ; Nasal Cavity/microbiology* ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S/genetics* ; Saliva/microbiology* ; Semen/microbiology* ; Vagina/microbiology*

ObjectiveTo investigate bacterial infection and the distribution of different bacterial species in the donor semen and the influence of different bacterial counts on semen quality.

METHODSBacterial colonies in the semen samples from 1 126 donors were counted with the Synbiosis Protocol 3 Automatic Colony Counter and the bacterial species with a colony count ≥10⁴ cfu/ml identified with the VITEK2 Compact Automatic Biochemical Analyzer. The Makler Sperm Counting Board was used to examine the semen quality of the semen samples with a colony count = 0 cfu/ml (n = 22, group A), those with a colony count <10⁴ cfu/ml (n = 22, group B) and those with a colony count ≥10⁴ cfu/ml (n = 22, group C). Univariate analysis was employed for comparison of semen quality among different groups.

RESULTSAmong the 1 126 donor semen samples cultured, 5 (0.44%) showed mixed bacterial contamination and 993 (88.58%) showed none but with growth of a certain species of bacteria, 2.22% (22/993) with a colony count ≥10⁴ cfu/ml, mainly including Streptococcus bovis, tiny bacilli, Staphylococcus epidermis, and Staphylococcus aureus, among which gram-positive and gram-negative bacteria accounted for 95.45% (21/22) and 4.54% (1/22), respectively. Compared with group A, groups B and C manifested significantly reduced total sperm count (［567.5 ± 327.6］ vs ［421.9 ± 155.9］ and ［389.9 ± 110.6］ × 106 per ejaculate, P <0.05) and percentage of progressively motile sperm (［65.0 ± 6.5］ vs ［61.0 ± 3.5］ and ［61.6 ± 4.3］ %, P <0.05). There were no statistically significant differences among the three groups in the semen liquefaction time, semen pH value, total sperm motility or percentage of morphologically normal sperm (P > 0.05). Of the 284 randomly selected semen samples, 34 (11.97%) were found positive for Ureaplasma urealyticum (UU) and no significant difference was observed in the semen quality between the UU-positive and UU-negative samples (P> 0.05).

CONCLUSIONSThe bacteria-positive rate is high in the donor semen and the bacterial species are varied, mainly including gram-positive bacteria. Semen quality is reduced with the increased number of bacterial colonies.

Analysis of Variance ; Bacteria ; classification ; isolation & purification ; Bacterial Load ; Humans ; Male ; Semen ; microbiology ; Semen Analysis ; Sperm Count ; Sperm Motility ; Spermatozoa ; Tissue Donors ; Ureaplasma urealyticum

OBJECTIVETo investigate the status of the semen of the infertility patients infected by Gardnerella vaginalis (Gv).

METHODSSemen samples from 373 clinic patients of infertility and vaginal samples from 63 positive patients' wives were collected from April 2002 to May 2003. And the samples were tested by nested polymerase chain reaction (nPCR).

RESULTSThe positive rate of the infertile males' semen infected by Gv was 44.2%, while that of the postive patients' wives was 87.3%.

CONCLUSIONThe positive rate of the infertile male's semen infected by Gv is high and Gv can be spread by sexual intercourse.

Adult ; Base Sequence ; Female ; Gardnerella vaginalis ; isolation & purification ; Humans ; Infertility, Male ; microbiology ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Semen ; microbiology

OBJECTIVETo investigate the impact of Staphylococcus aureus from infertile men on sperm motility and the relationship between virulence genes and the activity of spermatozoal immobilization.

METHODSWe collected 60 strains of non-repeated Staphylococcus aureus from the semen of 589 infertile males and analyzed the influence of Staphylococcus aureus on sperm motility using the computer-aided sperm analysis system. We selected the strains that apparently decreased sperm motility and detected their virulence genes by PCR.

RESULTSSperm motility was significantly decreased in 17 of the 60 strains of Staphylococcus aureus (P < 0.05). The main virulence genes in these strains were hlg (33.3%), scn (23.3%), cna (20%), hlb (20%), and clfA (18.3%), others including icaA, fnbA, tst, seb, hld, eta and sea. The scn gene carriers accounted for 47.1% in the spermatozal immobilization positive group, significantly higher than 14% in the negative group (P < 0.05). No statistically significant differences were found in the percentages of the carriers of the other virulence genes between the two groups (P > 0.05).

CONCLUSIONInfections of Staphylococcus aureus in male reproductive system can lead to the decrease of sperm motility, which may be associated with the Staphylococcus complement inhibitor encoding gene scn.

Humans ; Infertility, Male ; microbiology ; Male ; Polymerase Chain Reaction ; Semen ; microbiology ; Species Specificity ; Sperm Motility ; Staphylococcal Infections ; Staphylococcus aureus ; pathogenicity ; Virulence ; genetics

This case report describes a case of self-inflicted chronic bacterial keratoconjunctivitis involving the patient's own semen. A 20-year-old male soldier was referred to our clinic for the evaluation of refractory chronic bacterial conjunctivitis. Over the previous 4 months, he had been treated for copious mucous discharge, conjunctival injection, and superficial punctate keratitis in both eyes at an army hospital and a local eye clinic. Despite the use of topical and systemic antibiotics according to the results of conjunctival swab culture, there was no improvement. During the repeated smear and culture of conjunctival swabs, surprisingly, a few sperm were detected on Gram staining, revealing that the condition was self-inflicted bacterial keratoconjunctivitis involving the patient's own semen. Thus, in cases of chronic keratoconjunctivitis that do not respond to appropriate antibiotic treatment, self-inflicted disease or malingering should be considered.
Chronic Disease ; Conjunctiva/*injuries/microbiology/pathology ; Cornea/microbiology/*pathology ; Diagnosis, Differential ; Eye Infections, Bacterial/diagnosis/*etiology/microbiology ; Eye Injuries/*complications/diagnosis ; Humans ; Keratoconjunctivitis/diagnosis/*etiology/microbiology ; Male ; Self Mutilation/*complications/diagnosis ; *Semen ; Young Adult

INTRODUCTIONThis study aims to evaluate whether an increased polymorphonuclear leucocyte (PMN) count in semen is a good predictor of male genital tract infection, which is detected by semen culture.

METHODSA retrospective cross-sectional study examining the semen of 388 men was conducted at the in vitro fertilisation centre of a tertiary hospital. We compared the culture results of 109 men with increased semen PMN count against those of 279 men with normal semen PMN count.

RESULTSThere was no significant difference in the percentage of positive cultures between men with increased PMN count in their semen and those without PMN count elevation (original sensitivity 20.8%, specificity 70.3%; p = 0.1289). The overall percentage of positive semen cultures among all 388 patients was 18.6%.

CONCLUSIONBased on the positive cultures of significant organisms in the semen of our cohort, an increased semen PMN count is not a good predictor of genital tract infection in men.

Adult ; Bacterial Infections ; diagnosis ; microbiology ; Cross-Sectional Studies ; Humans ; Leukocyte Count ; Male ; Middle Aged ; Neutrophils ; cytology ; microbiology ; Reproductive Tract Infections ; diagnosis ; microbiology ; Retrospective Studies ; Semen ; cytology ; microbiology ; Sensitivity and Specificity ; Young Adult