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

Objective To analyze the infection status of human papilloma virus (HPV),Ureaplasma urealyticum (UU),Chlamydia trachomatis (CT),and Neisseria gonorrhoeae (NG) in clinical patients.Methods The laboratory specimens including urine,urethral swabs,and cervical swabs from 870 patients from January 1st 2014 to December 31st 2017 were retrospectively analyzed. HPV-DNA was detected by multiplex fluorescent PCR,and the UU-RNA,CT-RNA,and NG-RNA were determined by isothermal nucleic acid amplification. The positive rate of each pathogen and the distribution of positive rate between male and female patients were calculated. The samples were further divided into HPV-positive group and HPV-negative group,and the positive rates of UU-RNA,CT-RNA,and NG-RNA in these two groups were compared.Results The highest positive rate was 53.68%(467/870) for UU-RNA,followed by HPV-DNA [32.41%(282/870) ]and NG-RNA [2.18%(19/870)]. The total positive rate of high-risk (HR)-HPV(subtypes:16,18,31,33,35,39,45,51,52,56,58,59,and 68) [31.52%(209/663)]and UU in female patients [60.93%(404/663)] was significantly higher than that in male patients [17.39%(36/207),30.34%(63/207)](both P<0.001). The male patients had significantly higher CT positive rate in HR-HPV-positive group than in HR-HPV-negative group [22.58%(7/31) vs. 4.54%(8/176)](P<0.001). The female patients had significantly higher CT positive rate in HR-HPV-positive group than in HR-HPV-negative group [10.5%(21/200) vs. 5.61%(26/463)](P=0.024). The UU-RNA positive rate of females in the low-risk (LR)-HPV (subtypes:6 and 11) positive group was significantly higher than that in LR-HPV negative group [70.83%(34/48) vs.2.11%(13/615)](P<0.001).Conclusions Women are more susceptible to HR-HPV and UU infections. HR-HPV-positive patients are more likely to experience CT infection. In contrast,co-infection with UU is more common in LR-HPV-positive females.
Chlamydia Infections ; diagnosis ; epidemiology ; Chlamydia trachomatis ; isolation & purification ; Female ; Gonorrhea ; diagnosis ; epidemiology ; Humans ; Male ; Neisseria gonorrhoeae ; isolation & purification ; Papillomaviridae ; isolation & purification ; Papillomavirus Infections ; diagnosis ; epidemiology ; Retrospective Studies ; Ureaplasma Infections ; diagnosis ; epidemiology ; Ureaplasma urealyticum ; isolation & purification

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 study the postnatal changes in lymphocyte subsets in early preterm infants and the effect of perinatal factors on lymphocyte subsets.

METHODSA total of 61 early preterm infants were enrolled. Flow cytometry was used to measure the absolute counts of lymphocytes and lymphocyte subsets at 1, 7, 14, and 28 days after birth, as well as at 6 months after birth for 17 of these early preterm infants. The effects of perinatal factors, such as antepartum use of hormone, intrauterine infection, gestational age at birth, and Ureaplasma urealyticum (UU) colonization, on lymphocyte subsets were analyzed.

RESULTSThe absolute counts of lymphocyte subsets except natural killer (NK) cells were lowest at birth, increased rapidly at 1 week after birth, and reached the levels in healthy infants at 6 months; the count of NK cells remained at a low level and increased significantly at 6 months after birth. Compared with those with a gestational age of <28 weeks, the early preterm infants with a gestational age of ≥28 weeks had significantly higher absolute counts of T cells, T helper (Th) cells, and NK cells at 7 days after birth, a significantly higher absolute count of T cells at 14 days after birth, and significantly higher absolute counts of lymphocytes and Th cells at 28 days after birth (P<0.05). Compared with the group not using hormone, the group using hormone showed a significantly higher absolute count of T cells at 7 days after birth and significantly higher absolute counts of lymphocytes and all subsets at 14 days after birth (P<0.05). There was no significant difference in lymphocyte subsets at 1 day after birth between the intrauterine infection and non-infection groups (P>0.05); the intrauterine infection group had significantly higher absolute counts of B cells at 7 and 14 days after birth than the non-infection group. Compared those without UU colonization, the infants with UU colonization had significantly higher absolute counts of lymphocytes, T cells, Th cells, and Ts cells at 1 day after birth and a significantly higher absolute count of B cells at 14 days after birth.

CONCLUSIONSEarly preterm infants have deficiencies in innate immune cells at birth and normal levels at about 6 months after birth. Various perinatal factors including antepartum use of hormone, gestational age at birth, intrauterine infection, and UU colonization have long-term effects on lymphocyte subsets in early preterm infants.

Female ; Humans ; Infant, Newborn ; Infant, Premature ; immunology ; Lymphocyte Subsets ; microbiology ; physiology ; Male ; Ureaplasma urealyticum ; isolation & purification

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

The aim of this study was to determine whether intra-amniotic infection/inflammation (IAI) was associated with subsequent ruptured membranes in women with preterm labor and intact membranes who had a clinically indicated amniocentesis. This retrospective cohort study included 237 consecutive women with preterm labor (20-34.6 weeks) who underwent amniocentesis. The clinical and laboratory parameters evaluated included demographic variables, gestational age, C-reactive protein (CRP) and amniotic fluid (AF) white blood cell, interleukin-6 (IL-6) and culture results. IAI was defined as a positive AF culture and/or an elevated AF IL-6 level (>2.6 ng/mL). The primary outcome was ruptured membranes in the absence of active labor occurring within 48 hours of amniocentesis. Preterm premature rupture of membranes subsequently developed in 10 (4.2%) women within 48 hr of amniocentesis. Multivariate analysis demonstrated that only IAI was independently associated with the ruptured membranes occurring within 48 hr of amniocentesis. In the predictive model based on variables assessed before amniocentesis, only CRP level was retained. IAI is an independent risk factor for subsequent ruptured membranes after clinically indicated amniocentesis in preterm labor. Prior to amniocentesis, measurement of serum CRP level can provide a risk assessment for the subsequent development of ruptured membranes after the procedure.
Adult ; Amniocentesis/*adverse effects ; Amnion/physiopathology ; Amniotic Fluid/cytology/metabolism/microbiology ; Bacterial Infections/*etiology/microbiology ; C-Reactive Protein/analysis ; Cohort Studies ; Demography ; Female ; Gestational Age ; Humans ; Inflammation/*etiology ; Interleukin-6/metabolism ; Leukocytes/cytology ; Multivariate Analysis ; Mycoplasma/isolation & purification ; Obstetric Labor, Premature/*etiology ; Pregnancy ; ROC Curve ; Retrospective Studies ; Risk Factors ; Ureaplasma urealyticum/isolation & purification

BACKGROUND: To investigate the risk factors for vaginal infections and antimicrobial susceptibilities of vaginal microorganisms among women who experienced preterm birth (PTB), we compared the prevalence of vaginal microorganisms between women who experienced preterm labor (PTL) without preterm delivery and spontaneous PTB. METHODS: Vaginal swab specimens from 126 pregnant women who experienced PTL were tested for group B streptococcus (GBS), Mycoplasma hominis, Mycoplasma genitalium, Ureaplasma urealyticum, Chlamydia trachomatis, Trichomonas vaginalis, Neisseria gonorrhoeae, Treponema pallidum, herpes simplex virus (HSV) I and II, and bacterial vaginosis. A control group of 91 pregnant women was tested for GBS. Antimicrobial susceptibility tests were performed for GBS, M. hominis, and U. urealyticum. RESULTS: The overall detection rates for each microorganism were: U. urealyticum, 62.7%; M. hominis, 12.7%; GBS, 7.9%; C. trachomatis, 2.4%; and HSV type II, 0.8%. The colonization rate of GBS in control group was 17.6%. The prevalence of GBS, M. hominis, and U. urealyticum in PTL without preterm delivery and spontaneous PTB were 3.8% and 8.7% (relative risk [RR], 2.26), 3.8% and 17.3% (RR, 4.52), and 53.8% and 60.9% (RR, 1.13), respectively, showing no significant difference between the 2 groups. The detection rate of M. hominis by PCR was higher than that by culture method (11.1% vs. 4.0%, P=0.010). The detection rates of U. urealyticum by PCR and culture method were 16.7% and 57.1%, respectively. CONCLUSIONS: There was no significant difference in the prevalence of GBS, M. hominis, and U. urealyticum between the spontaneous PTB and PTL without preterm delivery groups.
Female ; Humans ; Microbial Sensitivity Tests ; Mycoplasma Infections/complications/microbiology ; Mycoplasma hominis/isolation & purification ; Obstetric Labor, Premature/*epidemiology/etiology ; Pregnancy ; Pregnancy Complications, Infectious/epidemiology/microbiology ; Premature Birth/*epidemiology/etiology ; Prevalence ; Risk Factors ; Streptococcal Infections/complications/microbiology ; Streptococcus agalactiae/isolation & purification ; Ureaplasma Infections/complications/microbiology ; Ureaplasma urealyticum/isolation & purification ; Vagina/*microbiology

OBJECTIVETo investigate the association of Ureaplasma urealyticum (UU) infection with the incidence of bronchopulmonary dysplasia (BPD), to compare the clinical manifestations and prognosis of BPD infants with or without Ureaplasma urealyticum infection.

METHODData were retrospectively collected between January 2004 and June 2011. All infants whose gestational age was ≤ 32 w and survived at 36 w were included in this study. Endotracheal aspirates were collected for UU polymerase chain reaction (PCR) within the first 48 hr of life. Statistical analyses were performed by using SPSS 11.5 software. The clinical characteristics of infants in the two groups were compared. The association of UU infection and BPD was analyzed and the clinical manifestations and prognosis of BPD in the two groups were compared.

RESULTThe results of PCR for UU were positive while that for other pathogens were negative in 168 infants whose chest X rays confirmed pulmonary inflammatory changes (UU group). The results of PCR for UU were negative in 393 infants (non-UU group). Except for premature rupture of membranes >24 hr, the rates of vaginal delivery, neonatal respiratory distress syndrome (NRDS) and surfactant use, there was no significant difference in the demographics and other baseline clinical characteristics of the two groups. The incidence of BPD was higher in UU group than in non-UU group and there was statistically significant difference in severity of BPD (P = 0.044, 0.031). The infants had been followed up until they were 1 year old. Compared to infants in non-UU group, infants in UU group showed no significant differences in the rate of death of pulmonary infection in moderate and severe BPD infants, the same as the rates of BPD infants hospitalized again or hospitalized more than 2 times because of pulmonary infection or/and wheezing episode in the first year after birth.

CONCLUSIONPreterm infants infected with UU were more likely to have BPD than non-UU infants. BPD infants associated with UU infection were more severe than that in non-UU infants. Prognosis of BPD infants associated with UU infection was similar to that of the infants whose BPD was not associated with UU infection.

Bronchopulmonary Dysplasia ; epidemiology ; etiology ; physiopathology ; Female ; Humans ; Infant ; Infant, Newborn ; Infant, Premature ; Infant, Premature, Diseases ; epidemiology ; etiology ; physiopathology ; Male ; Prognosis ; Retrospective Studies ; Risk Factors ; Severity of Illness Index ; Ureaplasma Infections ; complications ; epidemiology ; physiopathology ; Ureaplasma urealyticum ; isolation & purification

OBJECTIVETo study the relationship between microbial gene 16SrRNA and intrauterine infection.

METHODSThirty cases of single preterm birth were enrolled, including 16 cases due to premature rupture of membranes (PROM) (rupture time>18 hrs), 6 cases due to spontaneous preterm birth and 8 cases due to iatrogenic preterm birth. Ten cases of single term birth were used as the control group. Fetal membrane and placenta samples were obtained. Amniotic fluid, blood from cord or newborn babies as well as gastric fluid and tracheal secretions from infants with mechanical ventilation were also obtained. The histological features of placenta and fetal membranes were observed. Polymerase chain reaction (PCR) was used to detect the presence of microbial 16SrRNA and ureaplasma urealyticum (UU) in placenta, fetal membranes and other samples.

RESULTSTwenty-one (70%) cases were diagnosed as chorioamnionitis, characterized by neutrophil infiltration in fetal membrane and placenta tissues, especially in fetal membranes. Chorioamnionitis was most frequent in babies whose gestational age less than 32 weeks or birth weight lower than 1 500 g. Positive 16SrRNA gene was found in 12 cases, and positive UU gene in 10 cases in the preterm birth group. Neither 16SrRNA nor UU gene was detected in the control group. The PROM preterm babies developed more frequent infection than the babies premature born due to other causes, but there were no statistically significant differences in the incidence of infection.

CONCLUSIONSChorioamnionitis may be the major cause of PROM and premature birth. The detection of microbial genes is valuable in identification of intrauterine infection.

Chorioamnionitis ; diagnosis ; Female ; Fetal Membranes, Premature Rupture ; etiology ; Humans ; Infant, Newborn ; Infant, Premature ; Placenta ; microbiology ; pathology ; Pregnancy ; RNA, Ribosomal, 16S ; genetics ; Ureaplasma urealyticum ; genetics ; isolation & purification

OBJECTIVETo investigate the infection and the drug resistance status of mycoplasma and chlamydiae in genitourinary tracts of children with suspected nongonococcal urethritis (NGU) and provide information for clinical rational administration of antimicrobial agents.

METHODSSamples of genitourinary tract secretion from 146 children who were suspected of having nongonococcal urethritis or colpitis were collected and tested for mycoplasma via culture and for chlamydia with antigen detection. Meanwhile, susceptibility test was carried out on the samples which were positive in mycoplasma cultivation. Chlamydia antigen was detected by the polymer conjugate-enhanced (PCE) indirect enzyme immunoassay (EIA) (IDEIA PCE Chlamydia; DAKO). The mycoplasma culture medium was produced by Nanjing Liming Biological Products Co,. Ltd. Antibiotics used for susceptibility test were erythromycin, roxithromycin, josamycin, leucomycin, meleumycin, rovamycin, azithromycin, clarithromycin, cycloate erythromycin, and clindamycin.

RESULTSFifteen samples were positive for Chlamydia trachomatis (Ct) by antigen detection (10.3%), 82 samples were positive in mycoplasma cultivation (56.2%), and among the 82 samples, 58 were positive for Ureaplasma urealyticum (Uu, 39.7%), 9 were positive for Mycoplasma hominis (Mh, 6.2%), and 15 were positive for Uu and Mh (10.3%). Of all the samples, 4 were positive for both Uu and Ct (2.7%). The rates of drug resistance of the 10 commonly used antibiotics were as follows: erythromycin 32.9%, roxithromycin 41.5%, josamycin 19.5%, leucomycin 22.0%, meleumycin 28.0%, rovamycin 30.5%, azithromycin 37.8%, clarithromycin 26.8%, davercin 24.4%, and clindamycin 26.8%, respectively. The results indicated that drug resistance rates of josamycin and leucomycin were the lowest, and the rates of roxithromycin and azithromycin were the highest.

CONCLUSIONSThe infection rates of mycoplasma and chlamydia in children suspected NGU were high. Mycoplasma showed drug resistance to a different degree to 10 common antibiotics. The results of chemosensitivity showed that josamycin had the highest susceptibility rate.

Anti-Bacterial Agents ; pharmacology ; Child ; Child, Preschool ; Chlamydia Infections ; microbiology ; Chlamydia trachomatis ; drug effects ; isolation & purification ; Drug Resistance, Bacterial ; Female ; Humans ; Male ; Microbial Sensitivity Tests ; Mycoplasma Infections ; microbiology ; Mycoplasma hominis ; drug effects ; isolation & purification ; Ureaplasma Infections ; microbiology ; Ureaplasma urealyticum ; drug effects ; isolation & purification