Cheap and rapid in-house method for direct identification of positive blood cultures by MALDI-TOF MS technology.

BACKGROUND: Rapid and accurate pathogen identification in blood cultures is very important for septic patients and has major consequences on morbidity and mortality rates. In recent years, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based technology has become useful for highly specific and sensitive identification of bacteria and yeasts from clinical samples including sterile body fluids. Additional in-house methods enabled direct identification from blood cultures following various preparation protocols. METHODS: Blood culture (5 ml) was harvested from each positive bottle following growth identification by BACTEC™ FX system and transferred into a VACUETTE® Z Serum Sep Clot Activator tube containing an inert gel, which following centrifugation separates microorganisms from the blood cells. We used MALDI-TOF MS analysis for identification of microorganisms collected from the gel surface. RESULTS: Positive blood culture bottles (186) were collected. In comparison with the routine method, 99% (184/186) and 90% (168/186) of the isolates were correctly identified by the SepsiTyper kit and the in-house method, respectively. We found high concordance (Pearson coefficient = 0.7, p <  0.0001) between our in-house method and the SepsiTyper kit. Additionally, high correlation was found in sub-groups of identified bacteria, with Pearson coefficients of 0.77 (p <  0.0001), 0.67 (p <  0.0001), and 0.73 (p <  0.007) for Gram negative, Gram positive, and anaerobic bacteria, respectively. CONCLUSIONS: Our in-house method was found to be in good agreement with the SepsiTyper kit. Considering the low costs and the rapid and easy implementation of this procedure, we propose our in-house method for the direct identification of bacteria from blood cultures.

Bloodstream Infections Caused by Contaminants: Epidemiology and Risk Factors: A 10-Year Surveillance.

BACKGROUND: Skin colonization of microorganisms in blood cultures (BCs) are generally considered clinically non-significant and can be the source of a true infection, particularly in immunosuppressed patients. OBJECTIVES: To study the epidemiology and risk factors for bacteremia caused by contaminants. METHODS: This retrospective, descriptive study is based on adult BCs collected (2004-2013) and categorized as positive (True bacteremia [TrueB] or contamination) or negative. Clinical, demographic, and laboratory characteristics of BCs positive for the six most common potential contaminant pathogens (PCPs) that can cause TrueB and contamination (Coagulase-negative Staphylococcus [CoNS], Streptococcus viridans, Propionibacterium acnes, Corynebacterium spp., Bacillus spp., Clostridium spp.) were assessed. Ninety-two TrueB were identified vs. 196 contaminations (1:2 ratio). RESULTS: From 74,014 BCs, PCPs were found in 3735 samples, of which 3643 (97.5%) were contaminations and 92 (2.5%) were TrueB. The overall rate of BC contamination decreased during the study period from 6.7% to 3.8%. CoNS was the most common PCP. Bacillus spp. were only contaminants. Clostridium spp. and Streptococcus viridans were more often TrueB. In a multivariate model, predictors of TrueB included high creatinine levels, Streptococcus viridans in BC, and multiple positive BCs. A single culture of CoNS was strongly predictive of contamination. CONCLUSIONS: Ten years of data on BCs, focusing on six PCPs, demonstrates a significant, yet insufficient reduction in the rate of contamination. High creatinine level, isolation of Streptococcus viridans, and multiple positive BCs were predictors of TrueB, while growth of CoNS was strongly predictive of contamination. This model could assist in diagnostic and therapeutic decision making.

Microbiologic Data in Acute Cholecystitis: Ten Years' Experience from Bile Cultures Obtained during Percutaneous Cholecystostomy.

BACKGROUND: The aim of the study was to describe the microbiology and susceptibility patterns in acute cholecystitis by examining bile culture results from patients who underwent percutaneous cholecystostomy and examine concordance with empiric treatment. PATIENTS AND METHODS: A total of 124 patients with acute cholecystitis underwent percutaneous cholecystostomy between 2003 and 2012 at Emek Medical Center, Israel. Data on bile and blood culture results, isolate susceptibility, and clinical outcomes were retrieved from patient files. RESULTS: Bile cultures obtained from 116 patients were positive in 70 (60.3%) patients. Blood cultures obtained from 77 patients were positive in 23 (31.1%). Escherichia coli was the most common isolate in 28.6% of bile cultures and 43.5% of blood cultures. The concordance between empiric treatment coverage and culture isolate susceptibility was 67.6%. In most discordant cases, the isolates were Enterobacter spp. (40.9%) and Enterococcus spp. (31.8%). Overall, the in-hospital mortality rate was 7%: 2% in patients with concordant treatment compared with 14% in patients with discordant treatment (p = 0.09). Empiric antibiotic regimens were adequate in only two-thirds of patients. CONCLUSIONS: There might be a trend for poorer outcome in patients treated with inadequate antibiotic agents, emphasizing the importance of tailoring antibiotic treatment.

Trends in the Epidemiology of Campylobacteriosis in Israel (1999-2012).

The objective of this study was to examine the recent trends in the epidemiology of campylobacteriosis in Israel. A Sentinel Laboratory-Based Surveillance Network for Bacterial Enteric Diseases was established in Israel by the Israel Center for Disease Control (ICDC). This network generated data on subjects from whom Campylobacter spp. was isolated in community and hospital laboratories. Further characterization of the isolates was done at the Campylobacter National Reference Laboratory. Data from these two sources were integrated and analyzed at the ICDC. Between 1999 and 2012, 40,978 Campylobacter stool isolates were reported to the ICDC by the sentinel laboratories. The incidence rate of campylobacteriosis increased from 65.7 per 100,000 in 1999 to 101.7 per 100,000 in 2012. This increase resulted from a significant rise in the incidence of campylobacteriosis in the Jewish population which, since 2009, surpassed the consistent higher incidence of the disease in Israeli Arabs. The peak morbidity in Israel consistently occurred in late spring, with a risk excess in males compared with females, in younger age groups and earlier in the life span among Arabs than among Jews and others. These results suggest that further analytical studies should be carried out to identify risk factors responsible for the increased incidence of campylobacteriosis and better direct prevention and control of the disease in Israel.

Draft Genome Sequences of Streptococcus pneumoniae with High-Level Resistance to Respiratory Fluoroquinolones.

Streptococcus pneumoniaeis the leading cause of community-acquired pneumonia. Levofloxacin is a fluoroquinolone used for treatment of severe community-acquired pneumonia. Here, we describe the draft genome sequences ofS. pneumoniaewith emerging resistance to levofloxacin, resulting in failure of treatment of pneumococcal pneumonia.

Correlation between Clinical, Placental Histology and Microbiological Findings in Spontaneous Preterm Births.

AIMS: To examine the occurrence of chorioamnionitis and abruption among women who had a spontaneous preterm birth (SPTB), the correlation between clinical and placental findings, and the impact of these complications on neonatal outcome after delivery. METHODS: This was a retrospective case-control study conducted between 2008 and 2012 at a single teaching hospital. The study group included all women who had an SPTB (23-36 weeks). Placentas were cultured and underwent histological examination. RESULTS: A total of 478 women were included. The mean gestational age at delivery was 32.6 ± 3.1 weeks. Overall, 260 (54.4%) women had either clinical and/or histological abruption or chorioamnionitis. Clinical chorioamnionitis was diagnosed before birth in 14 (2.9%) women, while histological chorioamnionitis (HCA) in 84 (17.4%). Overall, 38 neonates had infection. Placental cultures were negative in 65.8% (25/38) of these neonates, and in 77.1% (27/38), HCA was ruled out. Logistic regression analysis revealed that neonatal morbidity and mortality were correlated with gestational age at delivery (p = 0.02), not with placental pathology (p = 0.08). CONCLUSIONS: Half of the women with PTB had clinical or histological abruption, chorioamnionitis or both. A partial correlation was found between clinical and placental findings. The main determinant of neonatal outcome was gestational age at delivery and not placental findings.

Reconsidering the Current Preterm Premature Rupture of Membranes Antibiotic Prophylactic Protocol.

OBJECTIVE: The purpose of our study was to determine whether the current antibiotic regimen for preterm premature rupture of membranes (PPROM) is adequate for covering the current causative agents and sensitivities of chorioamnionitis and early-onset neonatal sepsis. STUDY DESIGN: During a 3-year period, we retrieved the results from placental and amniotic membrane cultures obtained at delivery in cases of maternal fever, chorioamnionitis, and PPROM, and from blood cultures obtained from neonates with early-onset sepsis (EOS) in three participating hospitals. Sensitivity of pathogens to antimicrobial agents was performed using routine microbiologic techniques. RESULTS: There were 1,133 positive placental or amniotic cultures, 740 (65.3%) were from gram-negative Enterobacteriaceae. There were 27 neonates diagnosed with EOS with positive blood cultures. Aerobic Enterobacteriaceae accounted for 14 cases (52%) and group B streptococcus for 7 cases (26%). Of the Escherichia coli and Klebsiella sp., only 38% were sensitive to ampicillin. CONCLUSION: Local pathogens and their antibiotic sensitivity profiles should be explored every few years and an effective antibiotic protocol chosen to cover the main pathogens causing chorioamnionitis and EOS. Consideration should be made for changing ampicillin in women with PPROM to a regimen with better coverage of gram-negative Enterobacteriaceae.

Onychomycosis in Israel: epidemiological aspects.

Onychomycosis is a fungal infection treated orally for prolonged periods of treatment, caused primarily by Dermatophytes, Candida species and non-dermatophyte moulds (NDMs). The prevalence of specific aetiology may differ in dependence of environmental, geographic and demographic factors, and may affect management of the infection. The objective of this survey was to analyse epidemiologic parameters of onychomycosis in Israel. Data of a cohort of 27,093 patients were collected from six centres during a 2- and 10-year period. The diagnosis was based on microscopy of KOH/calcofluor mounts of nail scrapings and culture isolation. A positive result indicates isolation of a fungus in culture. Data were analysed for each centre and expressed as range for the whole cohort, using the spss v18 software. Analysis included three epidemiologic parameters: fungal aetiology in toe- and fingernails; association with gender; association with age group. Dermatophytes were the major causative agents and Trichophyton rubrum the most frequent isolate. Candida species were more frequent in women fingernails; frequency increased with age and C. parapsilosis the most frequent species. NDMs were isolated at low rate and Aspergillus terreus was the most frequent isolate. This is a first large cohort of onychomycosis patients from Israel analysed by defined epidemiological parameters.

Molecular characterization of methicillin-resistant Staphylococcus aureus strains isolated from patients with hospital readmissions.

It is unclear whether patients colonized with methicillin-resistant Staphylococcus aureus (MRSA) continue to harbor the same genotype during hospital readmissions. We characterized 140 MRSA strains isolated from 33 persistent MRSA carriers with hospital readmissions. Nearly half of the patients continued to harbor the same genotype, and the rest acquired different genotypes. Among 25 patients who received eradication therapy, 16 (64%) were colonized with MRSA strains exhibiting different genotypes from the preexisting one.

[Tuberculosis in pregnancy and puerperium].

According to the World HeaLth Organization, tuberculosis (TB) is the third leading cause of death worldwide among women at child bearing age. However, in Israel, a low TB prevalence country, TB in pregnant women is infrequent and infectious pulmonary TB at puerperium is rare. Early diagnosis of TB in pregnancy is challenging because the non-specific symptoms of early TB such as weakness, excess perspiration and tachycardia will usually be attributed to pregnancy. Furthermore, since health care givers attempt to avoid superfluous exposure of the fetus to radiation, and pregnant women are reluctant to be X-rayed, the diagnosis of active TB may be further delayed, especially if the woman is not in a risk group for TB. However, delaying treatment of TB in a pregnant woman, especially in advanced pregnancy may lead to TB in the fetus, TB infection of the new born transmitted from the mother who may also infect other mothers and their infants in the maternity ward. We report a case of highly infectious active TB diagnosed in a pregnant woman one week before delivery. The woman, a native Israeli, had no risk factor for TB except her recent stay in a high burden TB country. We present the diagnostic workup and therapeutic approach to the pregnant patient, the newborn infant and the measures applied to control infection. Awareness of risk factors for TB, the elusive symptoms of the disease during pregnancy, and implementing the necessary diagnostic workup at delivery is vital to minimize pregnancy related TB morbidity.

Are susceptibility tests enough, or should laboratories still seek ESBLs and carbapenemases directly?

Recent EUCAST advice asserts that, with low breakpoints, susceptibility results for cephalosporins and carbapenems can be reported 'as found', even for strains with extended-spectrum ß-lactamases (ESBLs) and carbapenemases. The CLSI has similar advice, but with higher ceftazidime and cefepime breakpoints than those of EUCAST. Pharmacodynamic and animal data are used to support these views, along with some analysis of clinical case series. We contend that such advice is misguided on three counts. First, whilst there are cases on record where cephalosporins and carbapenems have proved effective against infections due to low-MIC ESBL producers and low-MIC carbapenemase producers, respectively, there are similar numbers of cases where such therapy has failed. Second, routine susceptibility testing is less precise than in research analyses, meaning that ESBL and carbapenemase producers with 'real' MICs of 1-8 mg/L will oscillate between susceptibility categories according to who tests them and how. Third, although EUCAST continues to advocate ESBL and carbapenemase detection for epidemiological purposes, the likely consequence of not seeking these enzymes for treatment purposes is that some laboratories will not seek them at all, leading to a loss of critical infection control information. In short, it is prudent to continue to seek ESBLs and carbapenemases directly and, where they are found, generally to avoid substrate drugs as therapy.

Compliance of hospital staff with guidelines for the active surveillance of methicillin-resistant Staphylococcus aureus (MRSA) and its impact on rates of nosocomial MRSA bacteremia.

BACKGROUND: The compliance of hospital staff with guidelines for the active surveillance of methicillin-resistant Staphylococcus aureus (MRSA) in Israel has not been determined. OBJECTIVES: To evaluate the compliance of hospital staff with guidelines for the active surveillance of MRSA and assess its impact on the incidence of nosocomial MRSA bacteremia. METHODS: We assessed compliance with MRSA surveillance guidelines by assessing adherence to the screening protocol and reviewing medical and nursing charts of patients colonized with MRSA, and observed hand hygiene opportunities among health care workers and colonized patients. Rates of nosocomial MRSA bacteremia and of adherence with hand hygiene among overall hospital staff were obtained from archived data for the period 2001-2010. RESULTS: Only 32.4% of eligible patients were screened for MRSA carriage on admission, and 69.9% of MRSA carriers did not receive any eradication treatment. The mean rate of adherence to glove use among nurses and doctors was 69% and 31% respectively (P<0.01) and to hand hygiene 59% and 41% respectively (P<0.01). The hospital overall rate of adherence to hand hygiene increased from 42.3% in 2005 to 68.1% in 2010. Rates of nosocomial MRSA bacteremia decreased by 79.2%, from 0.48 (in 2001) to 0.1 (in 2010) per 1000 admissions (P<0.001). CONCLUSIONS: The compliance of medical and nursing staff with guidelines for active MRSA surveillance was poor. The encouraging increase in adherence to hand hygiene and concomitant decrease in nosocomial MRSA bacteremia is gratifying. The deficiencies in compliance with MRSA infection control policy warrant an adjusted strategy based on the hospital resources.

Cutaneous nocardiosis: report of two cases and review of the literature.

BACKGROUND: Cutaneous nocardiosis is an uncommon infectious disease that presents as a primary cutaneous infection or as a disseminated disease. It is often misdiagnosed because of its rarity and nonspecific clinical picture. METHODS: We report a case of each type. The first case is an immunocompetent patient who was infected by Nocardia while gardening and developed a superficial skin infection--one of the three clinical manifestations of primary cutaneous nocardiosis. The second case is an immunocompromised patient with pulmonary nocardiosis that extended to the skin as part of a disseminated disease. RESULTS: The immunocompetent patient with primary cutaneous nocardiosis had the classical features of a superficial skin infection. He had a nodular­pustular lesion on the right arm, which appeared 7 days after gardening with bare hands. Nocardia was identified in a skin culture taken from a pustule, unfortunately not to the species level. Treatment with minocycline for 3 months resulted in full remission of the lesion. The immunocompromised patient with disseminated nocardiosis had high fever, productive cough, hemoptysis, and erythematous nodules and pustules on the extremities. N. brasiliensis was isolated from bronchial samples and skin. Treatment with a high dose of trimethoprim and sulfamethoxazole for five months resulted in full recovery from cutaneous and pulmonary complaints. No relapse of the infection was found on follow-up in either patient. CONCLUSION: These cases demonstrate the need for a high degree of suspicion, focused clinical search, and appropriate laboratory procedures in the diagnosis and management of cutaneous nocardiosis.

Sole pathogen in acute bronchiolitis: is there a role for other organisms apart from respiratory syncytial virus?

BACKGROUND: Acute bronchiolitis (AB) is a common disease of young children with peak incidence during the winter season. Respiratory syncytial virus (RSV) is a major causative organism, yet recent relatively small sized studies have suggested an increased role of other organisms as sole or codetected organisms. The aim of this study was to assess the prevalence of sole- and mixed-organisms infections in hospitalized children with AB, using combined antigen-based and polymerase chain reaction assays (PCR). METHODS: Sputum or nasal wash specimens obtained from 490 previously healthy children < or =2 years of age hospitalized with AB between December 1, 2005 and March 31, 2006 were tested: (1) For RSV, by rapid antigen detection test; (2) For RSV, influenza A, B, Parainfluenza 1 to 3, and adenovirus antigens by direct fluorescent assay; (3) For influenza A and B, RSV, Parainfluenza 1 to 3 viruses RNA by reverse transcription (RT) PCR assay; (4) For human metapneumovirus and rhinovirus RNA by RT real-time PCR assay; (5) For adenovirus, and Bordetella pertussis DNA by conventional PCR assays; (6) For human bocavirus DNA by real-tine PCR assays. RESULTS: At least 1 organism was detected in 465 (91%) children. In 283 (61%), 117 (25%), and 23 (5%) children, 1, 2, and 3/4 organisms were detected, respectively. The most commonly detected organism was RSV, detected in 76%, and as a sole organism in 49%. Rhinovirus, human metapneumovirus, influenza virus A, bocavirus, Bordetella pertussis, and adenovirus were detected as a sole organism in 7%, 2.1%, 1%, 0.6%, 0.6%, and 0.2% of the children, respectively. CONCLUSIONS: Respiratory organisms were detected in the majority of the children, of whom about one third suffered from mixed organism infection. RSV was the most prevalent sole detected organism. The relevance of all other organisms may be much less than previously suggested.

Preoperative topical moxifloxacin 0.5% and povidone-iodine 5.0% versus povidone-iodine 5.0% alone to reduce bacterial colonization in the conjunctival sac.

PURPOSE: To assess the effectiveness of adding topical moxifloxacin 0.5% to topical povidone-iodine 5.0% for preoperative reduction of bacterial recovery from the conjunctiva. SETTING: Emek Medical Center, Afula, Israel. METHODS: The study population comprised adult patients scheduled for elective intraocular surgery. Two hours before surgery, patients were randomly assigned to topical therapy with moxifloxacin 0.5% drops (study group) or normal saline drops (control group). Povidone-iodine 5% solution was also instilled in the conjunctival sac in both groups for 3 minutes immediately before surgery. Conjunctival cultures were obtained before prophylactic therapy and just before surgery. The major outcome measures were the rate of bacterial colonization and bacterial type in conjunctival cultures obtained after instillation of povidone-iodine and immediately before surgery. RESULTS: Overall, 464 patients completed the study; there were 237 patients in the study group and 227 patients in the control group. Positive conjunctival cultures were obtained before prophylactic therapy in 91 patients (38%) in the study group and 94 patients (41%) in the control group and just before surgery in 10 patients (4%) and 6 patients (3%), respectively; neither difference was statistically significant. Coagulase-negative Staphylococcus was the most prevalent bacteria and was found equally in both groups. CONCLUSIONS: Treatment with povidone-iodine 5.0% alone was effective in preoperative reduction of conjunctival bacterial colonization. Adding topical moxifloxacin 0.5% to povidone-iodine 5.0% had no significant effect on further reduction in the bacterial colonization rate.

Pseudomonas stutzeri knee arthritis in a child: case report and review.

We report a child with Pseudomonas stutzeri-associated right knee arthritis following knife puncture wound. Only four children with P. stutzeri-associated infections have been reported in the English literature in the last 40 years of whom one suffered from calcaneal osteomyelitis caused by this pathogen. In both cases, the suggested mechanism of the infection was local rather than bacterium invasion.

Nontuberculous mycobacterial infections of the skin: A retrospective study of 25 cases.

BACKGROUND: Absence of a pathognomonic clinical picture and variable histologic findings often delay diagnosis of nontuberculous mycobacteria (NTM)-induced cutaneous infections, and antimicrobial therapy varies. OBJECTIVE: We sought to investigate the clinical, microbiologic, and pathological findings of cutaneous NTM infections and response to different treatments. METHODS: Records of patients with NTM infections of the skin confirmed by positive cultures or histologic findings were reviewed. Clinical, microbiologic, and epidemiologic data were collected and skin biopsy specimens were reassessed. RESULTS: The series included 25 cases, one diagnosed by histology and 24 by positive culture: 16 cases with Mycobacterium marinum, 3 of atypical Mycobacterium without species identification, and one each with M chelonae, M xenopi, M abscessus, M gordonae, and M fortuitum. One of 16 patients with M marinum developed tenosynovitis. Mean interval between clinical presentation and diagnosis was 7.1 months (range: 1-27.3 months). All isolates analyzed for antimicrobial susceptibility pattern were sensitive to clarithromycin. LIMITATIONS: Limitations include methods of case collection, retrospective study, and relatively small number of patients. CONCLUSIONS: Diagnosis of NTM should be confirmed by histology and bacteriology studies of tissue cultures. Strong clinical suggestion of M marinum infection warrants initial empirical treatment to prevent progression to deep infection. The recommended treatment as indicated by the results of the in vitro susceptibility and clinical response is clarithromycin.

Identification of the emerging pathogen Vibrio vulnificus biotype 3 by commercially available phenotypic methods.

Identification of the emerging pathogen Vibrio vulnificus biotype 3 has become a challenge for clinical laboratories in the last few years. In this study, the abilities of five commercial systems to identify this new species have been evaluated for the first time, using a unique collection of strains. Fifty-one well-documented wild strains of V. vulnificus biotype 3 were processed using API 20 NE, GNI+ Vitek 1 cards, ID-GNB Vitek 2 cards, Neg Combo 20 Microscan panels, and NMIC/ID-5 BD Phoenix panels. The numbers of strains identified as V. vulnificus by ID-GNB, NMIC/ID-5, and GNI+ were 50 (98.0%), 46 (90.2%), and 7 (13.7%), respectively. Neg Combo 20 Microscan panels and API 20 NE were unable to identify any of the strains of this emerging pathogen to the species level and mostly misidentifies them as other species of the Vibrionaceae family. Data on the phenotypic pattern of V. vulnificus biotype 3 when processed in all five systems as presented here could help clinical laboratories in identifying this new pathogen.

Actinomyces canaliculitis: diagnosis of a masquerading disease.

BACKGROUND: To review the diagnosis and management of seven cases of Actinomyces canaliculitis. METHODS: Culture of discharge was performed in six of seven patients with Actinomyces canaliculitis using a PD Plus/F blood culture bottle. All patients were treated by canaliculotomy with curettage of dacryoliths, followed by treatment with systemic penicillin and Sulphacetamide drops over a period of 3-6 months. Part of the curetted concretions was fixed on a glass slide and part was sent to the laboratory for culture. RESULTS: Four patients were women and three men with age ranging between 43 and 90 years. The average time lapse between onset of symptoms until diagnosis was 3 years. All cases presented with epiphora, chronic conjunctivitis, palpably thickened canaliculus, and yellow punctal discharge. Diagnosis was achieved by culture of discharge in three of six cases, culture of concretions in three of five cases, and staining of dacryoliths in all seven cases. Follow-up ranged between 12 and 48 months. The canaliculitis resolved completely and all patients have patent canalicula. CONCLUSIONS: Actinomyces canaliculitis presents with epiphora, chronic purulent conjunctivitis, a palpably thickened canaliculus, and yellow punctual discharge. In suspect cases canuliculotomy and curettage should be performed, although canalicular reconstruction is generally unnecessary. Culture of discharge and concretions using PD Plus/F blood culture medium gave improved results over accepted norms. Fixation of smeared concretions on a slide in alcohol is simple and is diagnostic of the disease. We recommend long-term systemic penicillin treatment in Actinomyces canaliculitis.

Bacterial colonization of the uterine cervix and success rate in assisted reproduction: results of a prospective survey.

BACKGROUND: Overgrowth of bacteria in the birth canal is associated with an increased risk of late miscarriage, preterm labour, post-partum endometritis and low birthweight. Conception rates in assisted reproduction treatments (ART) remain frustratingly low. We examined whether the nature of bacterial flora, found in the uterine cervical canal at embryo transfer, is associated with the rate of conception in ART. METHODS: We sampled for bacteriological culture the cervical canal of 204 patients who underwent embryo transfer. Of these, 139 (68%) were of fresh embryos, following recent vaginal oocyte retrieval and prophylactic antibiotic therapy, and 65 (32%) of frozen-thawed embryos, without any vaginal intervention in the preceding days. Bacteriological work-up included identification, colony count and antibiotic susceptibility profile. Conception was correlated with bacterial type and colony count. RESULTS: In 75 patients (36.8%) sterile cervical cultures or lactobacillus were recorded. Of these 75 patients, 23 (30.7%) conceived, whereas among the 129 in whom any pathogenic micro-organism was recovered only 21 (16.3%) conceived (P = 0.002). No difference in colonization was found between women who underwent frozen-thawed versus fresh embryo transfer (57 and 67% respectively). Any Gram-negative colonization was associated with no conception. All Gram-positive, and 90% of the Gram-negative bacteria, were sensitive to augmentin. CONCLUSIONS: Failure to conceive in ART is significantly associated with bacterial colonization of the uterine cervix.