This study primarily aimed to investigate the prevalence of human papillomavirus (HPV) and other common pathogens of sexually transmitted infections (STIs) in spermatozoa of infertile men and their effects on semen parameters. These pathogens included Ureaplasma urealyticum, Ureaplasma parvum, Chlamydia trachomatis, Mycoplasma genitalium , herpes simplex virus 2, Neisseria gonorrhoeae, Enterococcus faecalis, Streptococcus agalactiae, Pseudomonas aeruginosa , and Staphylococcus aureus . A total of 1951 men of infertile couples were recruited between 23 March 2023, and 17 May 2023, at the Department of Reproductive Medicine of The First People's Hospital of Yunnan Province (Kunming, China). Multiplex polymerase chain reaction and capillary electrophoresis were used for HPV genotyping. Polymerase chain reaction and electrophoresis were also used to detect the presence of other STIs. The overall prevalence of HPV infection was 12.4%. The top five prevalent HPV subtypes were types 56, 52, 43, 16, and 53 among those tested positive for HPV. Other common infections with high prevalence rates were Ureaplasma urealyticum (28.3%), Ureaplasma parvum (20.4%), and Enterococcus faecalis (9.5%). The prevalence rates of HPV coinfection with Ureaplasma urealyticum, Ureaplasma parvum, Chlamydia trachomatis, Mycoplasma genitalium , herpes simplex virus 2, Neisseria gonorrhoeae, Enterococcus faecalis, Streptococcus agalactiae , and Staphylococcus aureus were 24.8%, 25.4%, 10.6%, 6.4%, 2.4%, 7.9%, 5.9%, 0.9%, and 1.3%, respectively. The semen volume and total sperm count were greatly decreased by HPV infection alone. Coinfection with HPV and Ureaplasma urealyticum significantly reduced sperm motility and viability. Our study shows that coinfection with STIs is highly prevalent in the semen of infertile men and that coinfection with pathogens can seriously affect semen parameters, emphasizing the necessity of semen screening for STIs.
Humans ; Male ; Infertility, Male/epidemiology* ; Coinfection/microbiology* ; Papillomavirus Infections/virology* ; Adult ; Sexually Transmitted Diseases/complications* ; China/epidemiology* ; Staphylococcus aureus/isolation & purification* ; Chlamydia trachomatis/isolation & purification* ; Prevalence ; Mycoplasma genitalium/isolation & purification* ; Ureaplasma urealyticum/isolation & purification* ; Neisseria gonorrhoeae/isolation & purification* ; Enterococcus faecalis/isolation & purification* ; Streptococcus agalactiae/isolation & purification* ; Herpesvirus 2, Human/genetics* ; Pseudomonas aeruginosa/isolation & purification* ; Semen/virology* ; Sperm Motility ; Spermatozoa/microbiology* ; Human Papillomavirus Viruses

Ureaplasma urealyticum (UU) is one of the most commonly occurring pathogens associated with genital tract infections in infertile males, but the impact of seminal UU infection in semen on intrauterine insemination (IUI) outcomes is poorly understood. We collected data from 245 infertile couples who underwent IUI at The First Affiliated Hospital of USTC (Hefei, China) between January 2021 and January 2023. The subjects were classified into two groups according to their UU infection status: the UU-positive group and the UU-negative group. We compared semen parameters, pregnancy outcomes, and neonatal birth outcomes to investigate the impact of UU infection on IUI outcomes. There were no significantly statistical differences in various semen parameters, including semen volume, sperm concentration, total and progressive motility, sperm morphology, leukocyte count, the presence of anti-sperm antibody, and sperm DNA fragmentation index (DFI), between the UU-positive and UU-negative groups of male infertile patients (all P > 0.05). However, the high DNA stainability (HDS) status of sperm differed between the UU-positive and UU-negative groups, suggesting that seminal UU infection may affect sperm nuclear maturation ( P = 0.04). Additionally, there were no significant differences in pregnancy or neonatal birth outcomes between the two groups (all P > 0.05). These results suggest that IUI remains a viable and cost-effective option for infertile couples with UU infection who are facing infertility issues.
Humans ; Male ; Ureaplasma Infections/complications* ; Female ; Infertility, Male/therapy* ; Ureaplasma urealyticum/isolation & purification* ; Pregnancy ; Adult ; Pregnancy Outcome ; Semen Analysis ; Insemination, Artificial ; Semen/microbiology* ; China

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

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 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

Objective: To investigate the prevalence of Ureaplasma urealyticum (UU) infection in infertile men, its influence on routine semen parameters and the distribution of antisperm antibody (AsAb) and its types in infertile patients with UU infection. METHODS: We detected the positive rate of UU infection, semen parameters, and the distribution of AsAb and its types in 662 infertile men and 25 normal fertile male controls followed by comparison of the obtained data between the two groups of subjects. RESULTS: The positive rate of UU infection was significantly higher in the infertile men than in the normal controls (52.87% ［350/662］ vs 16.00% ［4/25］, χ2 = 11.68, P <0.05). The semen volume, sperm count, sperm concentration and percentage of progressively motile sperm were remarkably lower in the UU-positive infertile males than in the control group (P <0.05). No statistically significant difference was observed between the UU-positive and UU-negative groups in the positive rates of total AsAb (43.4% vs 36.5%, χ2 = 3.25, P >0.05) and AsAb IgA, IgM and IgG in the seminal plasma, or in the percentages of serum AsAb IgM (16.9% vs 20.5%, χ2 = 1.22, P >0.05) and IgG (32.7% vs 28.9%, χ2 = 0.99, P >0.05) except in that of serum AsAb IgA (23.6% vs 17.0%, χ2 = 4.03, P <0.05). CONCLUSIONS The UU infection rate is high in infertile males, which decreases the semen volume, total sperm count, motile sperm concentration and percentage of progressively motile sperm and increases the positive rate of serum AsAb IgA.
Antibodies, Bacterial ; analysis ; Humans ; Infertility, Male ; immunology ; microbiology ; Male ; Semen ; Sperm Count ; Spermatozoa ; immunology ; Ureaplasma Infections ; diagnosis ; immunology ; Ureaplasma urealyticum ; immunology

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

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 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

OBJECTIVETo investigate the semen quality and its influencing factors in preconception males in Nanjing area so as to provide some evidence for working out effective intervention measures.

METHODSTotally 687 men receiving preconceptional physical examination were enrolled in this study. A questionnaire survey was conducted among the subjects along with an analysis of their semen quality.

RESULTSThe median of sperm concentration was 63.3 x 10(6)/ml (95% CI [19.88-119] x 10(6)/ml). The median of grade a sperm was 33.03% (95% CI [19.38-55.05]%), that of grade a + b sperm was 52.08% (95% CI [39.53-69.37]%), and that of teratosperm was 91.75% (95% CI [69-100]%). The median concentration of seminal plasma PMN-elastase was 195.55 ng/ml (95% CI [76.16-3330.38] ng/ml) and that of seminal plasma zinc was 7.62 μmol/L (95% CI [1.5-23, 45] μmol/L). The positive rates of Ureaplasma urealyticum (UU), Chlamydia trachomatis (CT), and Gardnerella vaginalis (GV) were 42.4%, 0.3%, and 2.4%, respectively. The median of sperm DNA fragmentation index (DFI) of those whose wives had a history of adverse pregnancy was 20.25% (95% CI [2.15-68.25]%). Multivariate logistic regression analysis suggested that mental stress (OR 1.567, 95% CI [1.081-2.27]) and sedentariness (OR 1.772, 95% CI [1.211-2.592]) were independent risk factors for asthenospermia.

CONCLUSIONThe sperm quality of preconception males in Nanjing area is not encouraging, and it can be improved by changing undesirable lifestyle and reducing mental stress.

Adult ; Asthenozoospermia ; etiology ; China ; Chlamydia trachomatis ; isolation & purification ; DNA Fragmentation ; Gardnerella vaginalis ; isolation & purification ; Humans ; Leukocyte Elastase ; analysis ; Male ; Preconception Care ; Semen ; microbiology ; Semen Analysis ; statistics & numerical data ; Sperm Count ; statistics & numerical data ; Spermatozoa ; Ureaplasma urealyticum ; isolation & purification