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

To preliminarily understand the pathogenic mechanism of Mycoplasma genitalium (Mg) GroEL protein, we used bioinformatics tools to predict the structure and function of Mg GroEL protein and then constructed the recombinant plasmid pET-28a-GroEL. The protein expression was induced by 0.2 mmol/L IPTG, and the expressed protein was purified by Ni-iminodicitic acid (IDA) column affinity. Tohoku Hospital Pediatrics-1 (THP-1) cells were exposed to 2 μg/mL Mg rGroEL. The levels of interleukin (IL)-1β and tumor necrosis factor (TNF)-α in the cell supernatant were measured by ELISA, and that of IL-6 was measured by an automatic chemiluminescence instrument. The activation of the nuclear factor-kappa B (NF-κB) signaling pathway was visualized by immunofluorescence and Western blotting. The results showed that Mg GroEL was a stable hydrophilic protein composed of 543 amino acid residues, with the relative molecular mass of 58.44 kDa, an isoelectric point of 5.68, and a molecular formula of C₂₅₆₈H₄₃₀₀N₇₀₀O₈₂₅S₈. The secondary structure was mainly composed of α-helices and random coils. Mg GroEL contained 12 B-cell dominant epitopes and 10 T-cell dominant epitopes. It exhibited high homology with the GroEL proteins from Mycoplasma pneumoniae, M. agalactiae, M. arthritidis, M. hyopneumoniae, and M. bovis. Mg rGroEL activated the NF-κB signaling pathway and promoted the secretion of IL-1β, IL-6, and TNF-α in THP-1 cells. These results suggest that Mg GroEL exhibits substantial antigenicity and possesses the capability of triggering inflammation in host cells. This study establishes a theoretical basis for future investigations pertaining to the role and pathogenic mechanisms of Mg GroEL.
Mycoplasma genitalium/metabolism* ; Chaperonin 60/metabolism* ; Computational Biology ; Bacterial Proteins/genetics* ; Humans ; NF-kappa B/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Interleukin-1beta/genetics* ; Inflammation ; Interleukin-6/genetics* ; Recombinant Proteins/genetics* ; THP-1 Cells ; Signal Transduction ; Escherichia coli/metabolism*

Mycoplasma species (spp.) are bacteria that are difficult to detect. Currently, the polymerase chain reaction (PCR) is considered the most effective diagnostic tool to detect these microorganisms in both human and veterinary medicine. There are 13 known species of human Mycoplasma and 15 species of canine Mycoplasma. Owing to the difficulties in identifying the individual species of Mycoplasma, there is a lack of information regarding which species are saprophytic and which are pathogenic. The prevalence of the individual species is also unknown. In addition, in both humans and dogs, the results of some studies on the impact of Mycoplasma are conflicting. The presence of Mycoplasma spp. on the epithelium of reproductive tract is often associated with infertility, although they are also detected in healthy individuals. The occurrence of Mycoplasma spp. is more common in dogs (even 89%) than in humans (1.3%-4%). This is probably because the pH of a dog's genital is more conducive to the growth of Mycoplasma spp. than that of humans. Phylogenetically, human and canine Mycoplasma are related, and majority of them belong to the same taxonomic group. Furthermore, 40% of canine Mycoplasma spp. are placed in common clusters with those of human. This suggests that species from the same cluster can play a similar role in the canine and human reproductive tracts. This review summarizes the current state of knowledge about the impact of Mycoplasma on canine and human male fertility as well as the prospects of further development in this field.
Humans ; Dogs ; Male ; Animals ; Mycoplasma/genetics* ; Infertility ; Semen Analysis ; Polymerase Chain Reaction/methods* ; Prevalence ; Semen/chemistry*

OBJECTIVE: To study the drug resistance of METHODS: BALF specimens were collected from 245 children with RMPP who were admitted to the Children's Hospital Affiliated to Zhengzhou University from March 2016 to December 2020. A rapid cultured drug sensitivity assay was used to detect the resistance of MP isolates to nine commonly used antimicrobial drugs. The real-time PCR was used to measure MP DNA. The direct sequencing was used to detect gene mutations in MP 23SrRNA V region central ring. RESULTS: Among the 245 BALF specimens, 207 tested positive for MP DNA, with a positive rate of 84.5%. The results of drug susceptibility test showed that the children with RMPP had a resistance rate of > 70% to macrolide antimicrobial drugs, with the highest resistance rate to clarithromycin, followed by roxithromycin, clindamycin, acetylspiramycin, erythromycin, and azithromycin, and these children had a resistance rate of < 5% to quinolone antimicrobial drugs. Among the 207 MP DNA-positive specimens, 41 (19.8%) had no drug-resistance gene mutations and 166 (80.2%) had drug-resistance gene mutations, among which 154 (74.4%) had an A→G mutation at 2063 locus of 23SrRNA V region central ring, 7 (3.4%) had an A→G mutation at 2064 locus, and 5 (2.4%) had mutations in both 2063 and 2064 loci. Among the 166 specimens with point mutations of the MP 23SrRNA gene, 159 (95.8%) had point mutations at 2063 locus. The A→G point mutation at 2063 locus of 23SrRNA V region central ring had a great impact on resistance to macrolide antimicrobial drugs. There was a significant difference in the distribution of alleles at 2063 locus between the children with resistance to clarithromycin, roxithromycin, clindamycin, acetylspiramycin, erythromycin, and azithromycin ( CONCLUSIONS MP in the BALF of children with RMPP has a relatively high resistance rate to macrolide antimicrobial drugs. Resistance to macrolide antimicrobial drugs is closely associated with the A→G point mutation in the 23SrRNA gene, and the point mutation at 2063 locus of 23SrRNA V region central ring may affect the drug-resistance mechanism of MP.
Anti-Bacterial Agents/pharmacology* ; Bronchoalveolar Lavage Fluid ; Child ; Drug Resistance, Bacterial/genetics* ; Humans ; Mycoplasma pneumoniae/genetics* ; Pneumonia, Mycoplasma/drug therapy*

OBJECTIVEMutations in 23S rRNA gene are known to be associated with macrolide resistance in Mycoplasma pneumoniae (M. pneumoniae). However, these mutations alone do not fully explain the high resistance rates in Asia. The aim of this study was to investigate other possible mutations involved in macrolide resistance in M. pneumoniae.

METHODSThe whole genomes of 10 clinical isolates of M. pneumoniae with macrolide resistance were sequenced by Illumina HiSeq2000 platform. The role of the macrolide-specific efflux transporter was assessed by efflux-pump inhibition assays with reserpine and carbonyl cyanide m-chlorophenyl-hydrazone (CCCP).

RESULTSA total of 56 single nucleotide polymorphisms (SNPs) were identified in 10 clinical isolates in comparison to the reference strains M129 and FH. Strikingly, 4 of 30 SNPs causing non-synonymous mutations were clustered in macrolide-specific efflux system gene macB encoding macrolide-specific efflux pump protein of the ATP-binding cassette transporter family. In assays of the minimal inhibitory concentrations (MIC) of macrolide antibiotics in the presence of the efflux pump inhibitors caused a significant decrease of MICs, even under detectable levels in some strains.

CONCLUSIONOur study suggests that macrolide efflux pump may contribute to macrolide resistance in M. pneumoniae in addition to the common point mutations in 23S rRNA gene.

Anti-Bacterial Agents ; pharmacology ; Drug Resistance, Bacterial ; genetics ; Genome-Wide Association Study ; Macrolides ; pharmacology ; Microbial Sensitivity Tests ; Mutation ; Mycoplasma pneumoniae ; drug effects ; genetics

OBJECTIVETo investigate the association of drug resistance of Mycoplasma pneumoniae (MP) with DNA load and genotypes in children with MP pneumonia.

METHODSA total of 230 children who were hospitalized and diagnosed with MP pneumonia between January 2012 and December 2016 were enrolled. Throat swabs were collected from the 230 children, and a rapid drug sensitivity assay was used to determine the sensitivity of clinical isolates of MP to nine commonly used antibacterial agents. Quantitative real-time PCR was used to measure MP-DNA load in throat swabs. PCR sequencing was used to determine the genotype of 2063 locus of the MP 23S rRNA V domain.

RESULTSOf the 230 children, 86 (37.4%) had genotype A in 2063 locus, 134 (58.3%) had genotype G, 8 (3.5%) had genotype C, and 2 (0.9%) had genotype T. Mutant strains (genotype G+C+T) had a significantly higher MP-DNA load than wild-type strains (genotype A) (P<0.05). The strains resistant to erythromycin, azithromycin, clarithromycin, and clindamycin had a significantly higher MP-DNA load than non-resistant strains (P<0.05). MP had a high drug resistance rate to macrolide antibiotics. More than 60% of the cases with resistance to macrolides were found to have A2063G mutations. MP was rarely resistant to quinolones (less than 2%).

CONCLUSIONSMutations in 2063 locus of the MP 23S rRNA V domain may result in the resistance of MP to macrolides and the change in DNA load and can be used as a basis for selecting drugs for MP.

Child ; Child, Preschool ; DNA, Bacterial ; analysis ; Drug Resistance, Bacterial ; Female ; Genotype ; Humans ; Infant ; Male ; Mycoplasma pneumoniae ; drug effects ; genetics ; Pneumonia, Mycoplasma ; drug therapy ; microbiology

This study examined the occurrence of Anaplasma spp. and hemoplasma infection in leopard cats, Prionailurus bengalensis euptilurus, in Korea. Twenty-nine biological samples were tested by molecular analysis. Two (6.9%) and eight (27.6%) tested specimens were positive for Anaplasma bovis and hemoplasma infection, respectively. Based on our results, Anaplasma/Ehrlichia spp. and hemoplasma are regularly infecting leopard cat populations of Korea. Considering their endangered status, regular monitoring of infection by arthropod-borne pathogens known to cause clinical symptoms in feline hosts such as Anaplasma/Ehrlichia spp. and hemoplasma would be crucial as part of ongoing conservation efforts.
Anaplasma/*isolation & purification ; Anaplasmosis/*epidemiology/microbiology ; Animals ; DNA, Bacterial/genetics ; *Felidae ; Molecular Sequence Data ; Mycoplasma/*isolation & purification ; Mycoplasma Infections/epidemiology/microbiology/*veterinary ; Phylogeny ; Polymerase Chain Reaction/veterinary ; RNA, Ribosomal, 16S/genetics ; Republic of Korea/epidemiology ; Sequence Analysis, DNA/veterinary

The role of atypical bacteria and the effect of antibiotic treatments in acute bronchitis are still not clear. This study was conducted at 22 hospitals (17 primary care clinics and 5 university hospitals) in Korea. Outpatients (aged > or = 18 yr) who had an acute illness with a new cough and sputum (< or = 30 days) were enrolled in 2013. Multiplex real-time polymerase chain reaction (RT-PCR) was used to detect five atypical bacteria. A total of 435 patients were diagnosed as having acute bronchitis (vs. probable pneumonia, n = 75), and 1.8% (n = 8) were positive for atypical pathogens (Bordetella pertussis, n = 3; B. parapertussis, n = 0; Mycoplasma pneumoniae, n = 1; Chlamydophila pneumoniae, n = 3; Legionella pneumophila, n = 1). Among clinical symptoms and signs, only post-tussive vomiting was more frequent in patients with atypical pathogens than those without (P = 0.024). In all, 72.2% of the enrolled patients received antibiotic treatment at their first visits, and beta-lactams (29.4%) and quinolones (20.5%) were the most commonly prescribed agents. In conclusion, our study demonstrates that the incidence of atypical pathogens is low in patients with acute bronchitis, and the rate of antibiotic prescriptions is high.
Anti-Bacterial Agents/therapeutic use ; Bordetella parapertussis/genetics/*isolation & purification ; Bordetella pertussis/genetics/*isolation & purification ; Bronchitis/drug therapy/*microbiology ; Chlamydophila pneumoniae/genetics/*isolation & purification ; Community-Acquired Infections/microbiology ; Female ; Humans ; Hypertension/complications ; Legionella pneumophila/genetics/*isolation & purification ; Male ; Middle Aged ; Mycoplasma pneumoniae/genetics/*isolation & purification ; Real-Time Polymerase Chain Reaction ; Republic of Korea ; Sputum/microbiology

OBJECTIVEMycoplasma pneumoniae (MP) is an important pathogen for community-acquired pneumonia in children. MP infection was considered to be self-limited, but many severe refractory MP pneumonia cases have been reported in recent years. The reason for variation in severity of MP pneumonia remains unclear. MP virulence including drug-resistance and host immunologic function are important influencing factors. The present study aimed to clarify relationship between local MP load and severity of MP pneumonia.

METHODMP DNA was quantitatively detected by fluorescent real-time PCR in bronchoalveolar lavage fluid (BALF) from 77 children with MP pneumonia. They were classified into groups of low MP load ( < 10(3)/ml, n = 14) , moderate MP load (10(3)-10(6)/ml, n = 22) and high MP load ( > 10(6)/ml, n = 41) . Clinical symptoms, main laboratory and imaging results of children among the three groups were compared.

RESULTWhen compared with low load group and moderate load group, high load group had longer fever duration (7 d, 10 d vs. 12 d) , longer time to normalization of temperature with macrolide administration (4 d, 8 d vs. 10 d) , more patients with high fever (50.0%, 68.2% vs. 87.8%) and longer duration of fever than 10 d (35.7%, 50.0% vs. 73.2%).Statistically significant difference existed in CRP among the three groups (1.0 mg/L, 11.5 mg/L, 34 mg/L). Large field of consolidation or atelectasis were found in 58.5% of high load patients, much higher than 22.7% in moderate load and 14.3% in low load patients. Bilateral or massive pleural effusion was not found in low load group, while in moderate load and high load group, they were 13.6% and 24.4%. However, no significant difference was found in symptoms and main laboratory and imaging results among different age groups in high load patients.

CONCLUSIONThere is a close relationship between MP load in BALF and clinical characteristics in children with MP pneumonia. Those with high MP load have a more severe process.

Adolescent ; Bacterial Load ; Bronchoalveolar Lavage Fluid ; microbiology ; Child ; Child, Preschool ; Colony Count, Microbial ; DNA, Bacterial ; genetics ; Female ; Humans ; Infant ; Male ; Mycoplasma pneumoniae ; genetics ; isolation & purification ; Pneumonia, Mycoplasma ; microbiology ; pathology ; Real-Time Polymerase Chain Reaction ; Severity of Illness Index

Amino Acid Sequence ; Animals ; Bacterial Proteins ; genetics ; metabolism ; Bacterial Toxins ; genetics ; metabolism ; Base Sequence ; Community-Acquired Infections ; microbiology ; Gene Expression Regulation, Bacterial ; Humans ; Lung ; microbiology ; pathology ; Mycoplasma pneumoniae ; Pneumonia, Mycoplasma ; microbiology ; Respiratory Distress Syndrome, Adult ; microbiology