[Male genital tract infection: the point of view of the bacteriologist]. / Infection du tractus génital masculin: le point de vue du bactériologiste.

Male genital tract infection and inflammation have been associated to 8 to 35% of male infertility cases in various studies. Their investigation is part of a multi-disciplinary process including new techniques as DNA integrity study. Bacterial seminal infection can cause transient or chronic persistent inflammation, and the microbiological investigations, as well as leukospermia, secretory chlamydial IgA and inflammatory cytokines help to approach the responsibility of inflammation in infertility or pathological condition, leading to antibiotic and anti-inflammatory treatment. In Assisted Reproductive Techniques (ART), bacteriospermia must be eradicated for a safe semen preparation to inseminate or to fertilize oocytes. Leukocytes cannot be completely eliminated by sperm preparation and the presence of antibiotics and antioxydants in the culture media is questionned.

Circulating antibodies to a conserved epitope of the Chlamydia trachomatis 60 kDa heat shock protein (hsp60) in infertile couples and its relationship to antibodies to C.trachomatis surface antigens and the Escherichia coli and human HSP60.

To evaluate the relationship between immunity to specific regions of the Chlamydia trachomatis 60 kDa heat shock protein (hsp60), autoimmunity to human HSP60 and infertility, sera from 50 women and 45 men seen for an infertility evaluation were tested. Humoral immunity to human HSP60 was detected in 18% of women and 8.9% of men while antibodies to the Escherichia coli hsp60 were detected in 12% of women and 4.4% of men. These differences were not statistically significant. In contrast, antibodies to a synthetic peptide epitope of the chlamydial hsp60, encompassing amino acids 260-271 (chsp 260-271), were present in sera from 16 (32%) of the women but in only six (13.3%) of the men (P=0.03). Antibodies to chsp 260-271 were present in 11 out of 17 (64.7%) individuals with high titre (>1:160) immunoglobulin (Ig)G antibody to C.trachomatis surface antigens as opposed to only two out of 15 (13.3%) with low titre antibody and two out of of 17 (11.8%) with undetectable chlamydial antibody (P < 0.004). Antibodies to chsp 260-271 were also associated with humoral immunity to human HSP60. 50% of sera with, as opposed to only 18.6% of sera without, anti-human HSP60 IgG were positive for antibodies to chsp 260-271 (P=0.03). In contrast, there was no relationship found between immunity to the E.coli hsp60 and antibodies to human HSP60. Antibodies to chsp 260-271 were more prevalent in women with at least two spontaneous abortions (eight out of 13, 61.5%) than in women with other infertility diagnoses (six out of 35, 17.1%) (P=0.004). Thus, immunity to chsp 260-271 is more prevalent in women than in men, associated with autoimmunity to human HSP60 and may be an immunological marker for spontaneous abortion.

Pathogenesis of HSV and CMV Infections in Pregnancy.

Human herpesvirus (HHSV) and human cytomegalovirus (HCMV) infections during pregnancy are a major concern of public health because of the risk for severe sequelae for the fetuses and the neonates and because primary infections, reinfections and reactivations can be asymptomatic. The risk for neonatal herpes is mostly congenital, while the risk for HCMV infection is either prenatal or congenital. Screening exposed women has not brought definite solutions but is currently being evaluated. Among pregnant women with active infection, evaluation of the fetus for contamination and thus for the risk for severe immediate or long-term sequelae for neonates is the major goal. Diagnostic tools are available, cell culture still being the gold standard, and polymerase chain reaction (PCR) being currently evaluated for its contribution to diagnosis of active infection. Consensus for screening pregnant women as well as achievement of antiviral vaccines are the most urgent intervention strategies to develop in the near future.

[Comparative study of minimal inhibiting concentrations of doxycycline, ofloxacin and erythromycin against 18 recent isolates from Chlamydia trachomatis (1994-1995)]. / Etude comparative des concentrations minimales inhibitrices de la doxycycline, de l'ofloxacine et de l'érythromycine sur 18 isolats récents (1994-1995) de Chlamydia trachomatis.

In vitro activity of the 3 antimicrobial agents, against 18 recent isolates of Chlamydia trachomatis: 11 from urethral samples and 7 from endocervical samples, was tested by cell culture technique. HeLa 229 cells maintained in antibiotic-free MEM medium supplemented with 10% fetal calf serum and 2 mM glutamine were seeded into 96-well culture plates. After inoculation with 10(2) to 10(3) inclusion-forming units/ml of each strain of C. trachomatis, the culture medium was replaced by a maintenance medium containing 1 micrograms/ml of cycloheximide and serial two fold dilutions of the antibiotics. After 48 h incubation at 36 degrees C in 5% CO2 atmosphere, cells were fixed and inclusion bodies were stained using fluorescein-conjugated anti-Chlamydia monoclonal antibodies. MICs were defined as the lowest antibiotic concentrations required to inhibit the development of a single inclusion. MICs 90% (mg/l) were 0.054 for doxycyclin, 0.700 for ofloxacin and 0.150 for erythromycin. These results confirm the effective in vitro activity of the three antibiotics tested against C. trachomatis.

[Bacteria isolated in 1994-1995 in female upper genitalia infections and in male urethritis. Distribution and sensitivity to antibiotics]. / Bactéries isolées en 1994-1995 au cours des infections gynécologiques hautes et des uréthrites masculines. Distribution et sensibilité aux antibiotiques.

OBJECTIVE: Classify antibiotics according to their individual activity so as to identify those suitable for empiric therapy. METHODS: We studied bacterial strains isolated from patients with urethritis (n = 189) and upper genital tract infections (n = 163) between June 1994 and February 1995 in 3 hospital and 4 community laboratories. Upper genital tract infections were divided into two groups: proven infection on laparoscopy specimen (n = 79) and suspected infection with isolation of pathogen in cervical samples (n = 84). Pathogens isolated were: Chlamydia trachomatis in 36/12/15 cases respectively, Mycoplasma hominis in 12/20/13, Ureaplasma urealyticum in 55/30/15, Neisseria gonorrhoeae in 40/2/0, Haemophilus spp in 20/2/1, group B streptococci in 7/1/8, E. coli in 8/1/17 and miscellaneous in 11/8/15. The minimal inhibitory concentrations for all strains were determined in 4 laboratories for ofloxacin, erythromycin and doxycyclin against C. trachomatis, M. hominis and U. urealyticum, and for ofloxacin, erythromycin, doxycyclin, amoxicillin+clavulanate, cefotaxime and gentamicin against the other strains. The activity score (% susceptibility to each antibiotic weighted by the frequencies of each isolate in urethritis and upper genital tract infection based on recent French epidemiologic data) was calculated for each antibiotic. CONCLUSION: The antibiotics with the best empiric activity scores in urethritis were, in decreasing order: doxycyclin (90.4%), ofloxacin (88.1%), and erythromycin (50.2%). The most active combinations in upper genital tract infections were ofloxacin+amoxicillin (100%), doxycyclin+cefotaxime+metronidazole (95.9%) and doxycyclin+amoxicillin (95.3%).

Immune consequences of Chlamydia infections in pregnancy and in vitro fertilization outcome.

This review addresses the immune consequences of Chlamydial infections in pregnancy and in vitro fertilization (IVF) outcome. In pregnancy, many works have shown the risk for perterm labor, preterm birth, or miscarriage with current infection, and stress the need for screening and treatment early in pregnancy. IVF outcome needs to be studied in relation to specific markers like inflammatory cytokines and secretory antibodies to the 60 kD heat shock protein (hsp60) and to Chlamydia, to determine their ability to predict and influence pregnancy outcome.

Clinical consequences of immune response to CT upper genital tract infection in women.

C. TRACHOMATIS (CT) infections of the upper genital tract in women are either acute, sub acute or chronic. CT infection has a tendency to be chronic, latent and persistent as a consequence of the host immune reaction to CT major outer membrane protein, 57 Kd heat shock protein and lipopolysaccharide. Chlamydial persistence can be induced as a result of inflammatory and/or immune regulated cytokines, Interferon gamma depletion of tryptophan causes a stress response involving development of abnormal forms with increased levels of stress response proteins which maintain host immune responses with continuous fibrin exudate.The main clinical consequences are acute and chronic pelvic inflammatory disease, with infertility, ectopic pregnancy and, less frequently, chronic pelvic pain as late sequelae.PID, WHEN ACUTE, IS MARKED BY BILATERAL PELVIC PAIN, PLUS OTHER INFECTIOUS SIGNS IN TYPICAL CASES: fever, leucorrhea, red and purulent cervix. In 50% cases, infectious signs are slight or absent or there is an atypical clinical situation. Laparoscopy is the key for diagnosis. It allows the surgeon to have a direct look at the pelvic organs and perform microbiologic and histologic sampling. In severe cases, laparoscopy allows the surgeon to aspirate the purulent discharge and successfully treat pelvic abscesses.Chronic PID usually is clinically silent. It is in most cases discovered some years after the onset of CT infection, in women operated on for tubal infertility or ectopic pregnancy. Further studies, to evaluate treatments efficiency in chronic cases and factors leading to ectopic pregnancy or to recurrence, are indicated.

Post-therapeutic evolution of serum chlamydial antibody titers in women with acute salpingitis and tubal infertility.

OBJECTIVE: To determine if serologic data and, more particularly, antichlamydial immunoglobulin (Ig) M can be used for diagnosis of current chlamydial intrapelvic gynecologic infection. DESIGN: Forty-two women with acute salpingitis (group A), 131 women with tubal factor infertility (group B), and 98 pregnant women (control group C) were studied. SETTING: Hôpital Jean Rostand, Sèvres (patients), Laboratories Magenta and Eylau, Paris (serology), Institut Pasteur, Paris (cultures). INTERVENTIONS: Study groups: endocervical/urethral swabs, pelvic samples; serologic study before and after treatment. CONTROL GROUP: Serologic study. MAIN OUTCOME MEASURES: Serum samples were collected from each patient initially and 6 to 9 weeks later; additionally, two to five sequential sera were obtained from 22 (group A) and 25 (group B) patients with positive cultures, evolutive serology, or positive antichlamydial IgM. Sera were tested for antichlamydial IgG by a microimmunofluorescence assay using Chlamydia trachomatis elementary bodies and for IgA and IgM by whole inclusion-fluorescent assay. RESULTS: Before treatment, there was a correlation between the presence of antichlamydial IgM and positive cervical and/or intrapelvic chlamydia cultures. After treatment, antichlamydial IgM, when initially positive, rapidly disappeared in most subjects; its persistence after 4 months was significantly associated with tubal sequelae in group A patients and persistence of positive intrapelvic chlamydial cultures in group B women. CONCLUSION: Serologic analysis of women with acute salpingitis or tubal infertility, including antichlamydial IgM, may aid both in the before treatment diagnosis of chlamydial infection and in the follow-up evaluation.

Detection of Chlamydia trachomatis in fallopian tube tissue in women with postinfectious tubal infertility.

OBJECTIVE: Biopsy tissues from women with postinfectious tubal infertility were studied for the presence of Chlamydia trachomatis. STUDY DESIGN: Tubal biopsy specimens from 25 women with postinfectious tubal infertility undergoing laparoscopy for repair of fallopian tubes were evaluated by culture, in situ hybridization. Immunocytochemistry, and transmission electron microscopy for the presence of Chlamydia trachomatis. Serum was also tested for Chlamydia trachomatis antibodies. RESULTS: Chlamydia trachomatis was detected in postinfectious tubal biopsy specimens in three of 25 patients by culture, 12 of 24 by in situ hybridization, 15 of 22 by immunoperoxidase stain, and two of 10 by transmission electron microscopy. Serum antibody against Chlamydia trachomatis was detected in 15 of 21 patients. CONCLUSION: Chlamydia trachomatis deoxyribonucleic acid or antigens were detected at a high percentage (19/24 women) in the biopsy tissues of the fimbrial and peritubal adhesions by in situ hybridization or immunoperoxidase stain, suggesting a persistent infection in these women even after antibiotic treatment.

[The bacteriologic diagnosis of bacterial vaginosis in an urban practice]. / Le diagnostic bactériologique des vaginoses bactériennes en pratique de ville.

The term "bacterial vaginosis" carries both bacteriologic and clinical connotations. Indeed, it implies that the vagina is uninflamed (as shown by the absence of polymorphonuclear neutrophils), but that potential pathogens (particularly Gardnerella vaginalis) are present. Many studies conducted over the last 10 years have shown that a tentative diagnosis is simple to make, based on the sniff test, the aspect of the vaginal discharge, the presence of clue cells, and the absence of polymorphonuclear cells and lactobacilli on fresh smears. However, classical treatment with metronidazole can be insufficient given the innumerable microbial associations, and a more thorough bacteriological diagnosis is required. The presence of anaerobes is suggested by a positive sniff test. Direct examination reveals small Gram-negative rods in clue cells, but cannot differentiate between commensal atypical lactobacilli and Gram-positive anaerobes. Culture with CO2 supplementation or in anaerobic conditions yields Bacteroides and Mobiluncus spp, frequently associated with Gardnerella vaginalis and pathogenic Fusobacterium spp. Culture in appropriate conditions will detect metronidazole-resistant Propionibacteria and pathogenic anaerobic cocci. Antibiotic susceptibility testing of these organisms is costly and time-consuming, but can avoid therapeutic failures. Gardnerella vaginalis is also frequently associated in bacterial vaginosis with endogenous (mainly E. coli and group B streptococci) and Ureaplasma. Antibiotic susceptibility testing is useful if the predominant members of the flora are Gram-negative aerobes, group D streptococci or pathogenic anaerobes. Bacterial vaginosis is simple to diagnose but poses a real therapeutic problem since some cases call for a simple readjustment of the commensal flora while others require full chemotherapy.(ABSTRACT TRUNCATED AT 250 WORDS)

[Value of prostatic massage for detection of Chlamydia trachomatis in the male urethra]. / Intérêt du massage prostatique pour la mise en évidence de Chlamydia trachomatis dans l'urètre masculin.

The aim of this work was to evaluate prostatic massage (PM) as diagnostic method of chronic chlamydial infection (possible etiologic agent of male infertility or chronic prostatitis). 105 men were examined. One urethral swabbing and one urine sampling are done before PM. After PM, prostatic fluid is collected spontaneously and in a minimum volume of urine. Chlamydial isolation samples are treated by 3 methods: culture on HeLa 229 cells, immunoenzymology and direct immunofluorescence. Of 107 samplings, 22 were positive. In 13 patients (14 samplings) the result was positive by PM alone. In those cases, prostatic massage was the only way to establish the Chlamydial infection, when urethral swabbing, usual method, is negative. If confirmed, the interest of prostatic massage rests on detection of unknown genital Chlamydial infection, and this, by a non surgical method.