Objective: Pathogenic airway bacteria colonizing the neonatal airway increase the risk of childhood asthma, but little is known about the determinants of the establishment and dynamics of the airway microbiota in early life. We studied associations between perinatal risk factors and bacterial richness of the commensal milieu in the neonatal respiratory tract. Methods: Three hundred and twenty-eight children from the Copenhagen Prospective Studies on Asthma in the Childhood2000 (COPSAC2000) at-risk birth cohort were included in this study. The bacterial richness in each of the nasopharynxes of the 1-month old, asymptomatic neonates was analyzed by use of a culture-independent technique (T-RFLP). Information on perinatal risk factors included predisposition to asthma, allergy and eczema; social status of family; maternal exposures during pregnancy; mode of delivery; and postnatal exposures. The risk factor analysis was done by conventional statistics and partial least square discriminant analysis (PLSDA). Results: The nasopharyngeal bacterial community at 1-month displayed an average of 35 (IQR: 14–55, range 1–161) phylogenetically different bacteria groups. Season of birth was associated with nasopharyngeal bacterial richness at 1-month of age with a higher bacterial richness (p = 0.003) and more abundant specific bacterial profiles representing Gram-negative alpha-proteobacteria and Gram-positive Bacilli in the nasopharynx of summer-born children. Conclusion: Early postnatal bacterial colonization of the upper airways is significantly affected by birth season, emphasizing a future focus on the seasonality aspect in modelling the impact of early dynamic changes in airway bacterial communities in relation to later disease development.

The human vagina harbor a rich microbiota. The optimal state is dominated by lactobacilli that help to maintain health and prevent various diseases. However, the microbiota may rapidly change to a polymicrobial state that has been linked to a number of diseases. In the present study, the temporal changes of the vaginal microbiota in patients treated for sexually transmitted diseases or bacterial vaginosis (BV) and in untreated controls were studied for 26 days. The patients included 52 women treated with azithromycin, tetracyclines or moxifloxacin for present or suspected infection with Chlamydia trachomatis or Mycoplasma genitalium. Women with concurrent BV were also treated with metronidazole. The controls were 10 healthy women of matching age. The microbiota was analyzed by 16S rRNA gene deep sequencing, specific qPCRs and microscopy. There was generally good correlation between Nugent score and community state type (CST) and qPCR confirmed the sequencing results. By sequencing, more than 600 different taxa were found, but only 33 constituted more than 1 ‰ of the sequences. In both patients and controls the microbiota could be divided into three different community state types, CST-I, CST-III and CST-IV. Without metronidazole, the microbiota remained relatively stable regarding CST although changes were seen during menstrual periods. Administration of metronidazole changed the microbiota from CST-IV to CST-III in approximately 50% of the treated patients. In contrast, the CST was generally unaffected by azithromycin or tetracyclines. In 30% of the BV patients, Gardnerella vaginalis was not eradicated by metronidazole. The majority of women colonized with Ureaplasma parvum remained positive after azithromycin while U. urealyticum was eradicated.

No aetiology is found in up to 40% of men with symptomatic urethritis. Male partners of women with bacterial vaginosis (BV) may be at higher risk of non-gonococcal urethritis (NGU). The aim of this study was to examine the role of BV associated bacteria in first-void urine (FVU) in 97 asymptomatic men without urethritis (controls) and 44 men (cases) with NGU including 20 men with idiopathic urethritis (IU) attending a Swedish STD-clinic between January and October 2010. BV-associated bacteria and ureaplasmas were detected by quantitative PCR assays. All BV associated bacteria, except Megasphaera-like type 1, were strongly positively correlated with U. urealyticum p<0.005 and even stronger with the combined U. urealyticum and U. parvum load (p<0.0005) suggesting that ureaplasma induced elevated pH may stimulate the growth of BV associated bacteria. No statistically significant differences were found between IU cases and controls in the prevalence or load of BV associated bacteria or ureaplasmas. In multiple logistic regression, Megasphaera-like type 1 was associated with IU (p = 0.03), but most positive FVU samples contained very few bacteria and the finding may not be clinically relevant.

Metamizol wird in vielen Ländern häufig perioperativ eingesetzt. Unsicherheit besteht jedoch hinsichtlich der möglichen Komplikation einer Agranulozytose. Bei fehlender Evidenz aus der Literatur hat eine Arbeitsgruppe Expertenempfehlungen zum perioperativen Einsatz von Metamizol erarbeitet und in einem strukturierten formalen Konsensusprozess verabschiedet. Anschließend wurden die Empfehlungen in den Präsidien der Fachgesellschaften beraten und konsentiert. Die Expertengruppe stimmt überein, dass Blutbildkontrollen zur Überwachung der Metamizoltherapie beim kurzfristigen perioperativen Einsatz und bei Patienten ohne entsprechende Risikofaktoren für eine Neutropenie kein Standard sein sollen. Medizinisches Personal soll über die Symptome einer Agranulozytose und das Vorgehen bei Verdacht auf eine Agranulozytose informiert sein. Mit einer Risikoaufklärung soll der Patient über die Gabe von Metamizol, das Nutzen-Risiko-Verhältnis und mögliche Alternativen aufgeklärt werden. Andere Nichtopioidanalgetika werden von der Expertengruppe hinsichtlich Nutzen und Risiken nicht günstiger eingestuft als Metamizol. Eine Sicherungsaufklärung soll erfolgen, wenn über einige Tage Metamizol verabreicht wurde und/oder Patienten mit einer laufenden Metamizolmedikation aus stationärer oder ambulanter Behandlung entlassen werden, da sich eine Agranulozytose auch einige Tage nach Absetzen von Metamizol manifestieren kann. Weitere Empfehlungen betreffen die Information des weiterbehandelnden Arztes und die Vermeidung einer Reexposition bei stattgehabter metamizolbedingter Blutbildveränderung. Die Empfehlungen der Expertengruppe sollen das medizinische Personal und die Patienten für eine adäquate perioperative Anwendung von Metamizol sensibilisieren.

Background: Dipyrone (metamizole) is widely used for perioperative pain management in countries where it is marketed. However, uncertainty exists concerning the safe use of this drug, specifically considering the rare adverse event of an agranulocytosis. Methods: As evidence from published studies was lacking, an expert panel developed recommendations for the perioperative use of dipyrone. After a formal, structured consensus process, the recommendations were approved by the involved medical societies. Results: The panel agreed that blood cell counts shall not be standard for short-term perioperative use in patients unless they are at risk for neutropenia. The medical staff shall be aware of the symptoms and course of action when agranulocytosis is suspected. Patients shall be informed about the risks and benefits of dipyrone and about potential alternatives. The expert group concluded that dipyrone has a relatively positive risk-benefit ratio compared to other non-opioid analgesics. The group strongly recommended educating patients about the symptoms of agranulocytosis if they have received dipyrone over several days and/or treatment is to be continued after discharge, because agranulocytosis can occur several days after discontinuation of metamizole. Further recommendations refer to the information of the physician taking over the patient's care after discharge and the avoidance of re-exposure in patients having previously suffered from dipyrone-induced agranulocytosis. Conclusion: The group's recommendations shall be communicated in order to raise medical staff's and patients' awareness of the appropriate use of dipyrone in the perioperative period.

HAL is a multi-disciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés. Development of a multiplex PCR test for identification of serovars 1, 7, and 12 Øystein Angen, Peter Ahrens, Stine G. Jessing

Background Non-gonococcal urethritis (NGU) is a common syndrome in men. NGU may have several causes, but many cases are caused by sexually transmitted infections that may also cause complications in their female partners. Chlamydia trachomatis and Mycoplasma genitalium are the most common causes of NGU, but in up to 35% of the cases, none of the known viral or bacterial causes are found. Traditionally, pathogens have been detected using various culture techniques that may not identify all species present in the urethra. To address this, we used culture-independent methods for analysis of the male urethral microbiota. Methods This case-control study analysed first void urine samples, collected at STD clinics in Stockholm, Sweden from men with idiopathic urethritis (IU), i.e. negative for Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium, Ureaplasma urealyticum, Trichomonas vaginalis, adenovirus, and herpes simplex virus type 1 and -2 together with samples from men without urethritis. Forty-six controls and 39 idiopathic urethritis patients were analysed. Results The microbiota was highly diverse: None of the 302 operational taxonomic units (OTUs) found in negative controls and IU patients were found in all of the samples or even in all of the samples in one group. More than 50% of the OTUs were only found in one or two of the total of 85 samples. Still the most dominant 1/6 of the genera constituted 79% of the sequences. Hierarchical clustering in a heatmap showed no specific clustering of patients or controls. A number of IU patient samples were dominated by a single genus previously related to urethritis (Gardnerella, Haemophilus, Ureaplasma). Conclusion The male urethra contain a very diverse composition of bacteria, even in healthy controls. NGU may be caused by a number of different bacteria but more studies including a higher number of samples are needed for elucidation of the role of each species.

The aetiology of non-gonococcal urethritis (NGU) remains unexplained in 30-40% of patients. Urine samples from men attending Swedish sexually transmitted disease clinics were examined by species-specific quantitative PCRs for Chlamydia trachomatis, Mycoplasma genitalium, Trichomonas vaginalis, Ureaplasma urealyticum, U. parvum, adenovirus, herpes simplex virus, Neisseria meningitidis, Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pneumoniae. A total of 187 men with acute NGU (symptoms ≤ 30 days) and 24 with chronic NGU (symptoms < 30 days) were cases, and 73 men without NGU were controls. Number of lifetime sexual partners was negatively associated with U. urealyticum bacterial load. C. trachomatis and M. genitalium were associated with NGU, as was U. urealyticum, with bacterial loads ≥ 1.3 × 103 genome equivalents/ml urine. Virus and H. influenzae might explain a few NGU cases, but the aetiology in at least 24% of patients with acute NGU was unexplained. In multivariate analysis, detection of U. urealyticum was significantly more common in acute NGU (20%) compared with controls (11%).

Introduction: Continuous or episodic allergen exposure is a major risk factor of frequent symptoms and exacerbations for patients with allergic asthma. It has been shown that temperature-controlled laminar airflow (TLA) significantly reduced allergen exposure and airway inflammation and improved quality of life of patients with poorly controlled allergic asthma.Objective: The objective was to evaluate the effects of nighttime TLA when used during real-life conditions for 12 consecutive months in addition to the patients' regular medication.Methods: This multicenter, pre- and postretrospective observational study included patients with inadequately controlled moderate-to-severe allergic asthma who received add-on treatment with TLA for 12 consecutive months. Data on medication use, asthma control, asthma symptoms, lung function, use of hospital resources, and exacerbations were collected after 4 and 12 months and compared with corresponding data collected retrospectively from medical records during the year prior to inclusion in the study.Results: Data from 30 patients (mean age 28; range 8 - 70) completing 4 months and 27 patients completing 12 months of TLA use are presented. The mean number of exacerbations was reduced from 3.6 to 1.3 (p < 0.0001), and the ratio of asthma-related emergency room visits or hospitalizations diminished from 72.4 to 23.3% (p = 0.001) or from 44.8 to 20.0% (p < 0.05), respectively, after 12 months of TLA use. The Asthma Control Test index increased from 14.1 to 18.5 (p < 0.0001). After 4 months of TLA use, clear improvements can be shown for most variables in line with the data collected after 12 months.Conclusions: The addition of TLA to the patients' regular medication significantly reduced exacerbations, asthma symptoms, and the utilization of hospital resources. The data support that TLA may be an important new non-pharmacological approach in the management of poorly controlled allergic asthma.

A novel multiplex quantitative real-time polymerase chain reaction (qPCR) for simultaneous detection of U. urealyticum and U. parvum was developed and compared with quantitative culture in Shepard's 10 C medium for ureaplasmas in urethral swabs from 129 men and 66 women, and cervical swabs from 61 women. Using culture as the gold standard, the sensitivity of the qPCR was 96% and 95% for female urethral and cervical swabs, respectively. In male urethral swabs the sensitivity was 89%. The corresponding specificities were 100%, 87% and 99%. The qPCR showed a linear increasing DNA copy number with increasing colour-changing units. Although slightly less sensitive than culture, this multiplex qPCR assay detecting U. urealyticum and U. parvum constitutes a simple and fast alternative to the traditional methods for identification of ureaplasmas and allows simultaneous species differentiation and quantitation in clinical samples. Furthermore, specimens overgrown by other bacteria using the culture method can be evaluated in the qPCR.

Bacterial vaginosis (BV) is traditionally diagnosed using vaginal samples. The aim of this study was to investigate whether BV can be diagnosed from first-void urine (FVU). Self-collected vaginal smears, vaginal swabs, and FVU were obtained from 176 women. BV was diagnosed by Nugent's criteria. The FVU and vaginal swabs were analyzed by quantitative PCRs (qPCRs) for selected vaginal bacteria (Atopobium vaginae, Prevotella spp., Gardnerella vaginalis, bacterial vaginosis-associated bacterium 2, Eggerthella-like bacterium, "Leptotrichia amnionii," Megasphaera type 1), and all had an area under the receiver operating characteristic (ROC) curve of >85%, suggesting good prediction of BV according to the Nugent score. All seven bacteria in FVU were significantly associated with BV in univariate analysis. An accurate diagnosis of BV from urine was obtained in this population by a combination of qPCRs for Megasphaera type 1 and Prevotella spp. The same two bacteria remained significantly associated with BV in a multivariate model after adjusting for the other five species. There was no statistically significant difference between the sensitivities and specificities of BV diagnosis by molecular methods performed on swabs and FVU samples. A linear regression analysis showed good agreement between bacterial loads from swabs and FVU, but Prevotella spp. could be detected in high numbers in a few FVU samples without being present in swabs. This method will allow diagnosis of BV in studies where only urine has been collected and where detection of BV is considered relevant.

Die primäre ziliäre Dyskinesie (PCD) ist eine seltene angeborene Erkrankung der Zilien, die sich zumeist im respiratorischen System manifestiert.

BACKGROUND:Bacterial vaginosis (BV) is a common condition, although its aetiology remains unexplained. The aim of this study was to analyse the composition of vaginal microbiota in women from Greenland to provide a quantitative description and improve the understanding of BV.METHODS:Self-collected vaginal smears and swabs were obtained from 177 women. The vaginal smears were graded for BV according to Nugent's criteria. The vaginal swab samples were analysed by 19 quantitative PCRs (qPCRs) for selected vaginal bacteria and by PCR for four sexually transmitted infections (STIs).RESULTS:STIs were common: Mycoplasma genitalium 12%, Chlamydia trachomatis 7%, Neisseria gonorrhoeae 1%, and Trichomonas vaginalis 0.5%. BV was found in 45% of women, but was not associated with individual STIs. Seven of the 19 vaginal bacteria (Atopobium vaginae, Prevotella spp., Gardnerella vaginalis, BVAB2, Eggerthella-like bacterium, Leptotrichia amnionii, and Megasphaera type 1) had areas under the receiver operating characteristic (ROC) curve > 85%, suggesting they are good predictors of BV according to Nugent. Prevotella spp. had the highest odds ratio for BV (OR 437; 95% CI 82-2779) in univariate analysis considering only specimens with a bacterial load above the threshold determined by ROC curve analysis as positive, as well as the highest adjusted odds ratio in multivariate logistic regression analysis (OR 4.4; 95% CI 1.4-13.5). BV could be subdivided into clusters dominated by a single or a few species together.CONCLUSIONS:BV by Nugent score was highly prevalent. Two of seven key species (Prevotella spp. and A. vaginae) remained significantly associated with BV in a multivariate model after adjusting for other bacterial species. G. vaginalis and Prevotella spp. defined the majority of BV clusters.

Background Little is known about the presence of bacterial vaginosis (BV) associated bacteria in men, but male partners of women with BV have been reported to have a high risk of urethritis. We aimed to examine the role of BV associated bacteria in urine specimens from men with and without non-gonococcal urethritis (NGU). Methods First-pass urines were collected from 44 men with symptomatic NGU (≥ 5 PMNL/hpf) and 97 asymptomatic men without NGU (< 5 PMNL/hpf). Samples were tested for Chlamydia trachomatis(Ct), Mycoplasma genitalium(Mg), Ureaplasma urealyticum (Uu), U. parvum(Up), HSV 1 and 2, and adenovirus by PCR. Quantitative PCRs were performed to detect Gardnerella vaginalis, BVAB 2, Eggerthella-like uncultured bacterium, Megasphaeratype 1 , Leptotrichia amnionii, Atopobium vaginae, Sneathia sanguinegens, and Prevotella sp. Results Ct was detected in 9 (21%) cases with NGU and 1 (1%) control without NGU. Mg was detected in 10 cases (23%) and none of the controls. Corresponding figures were for Uu 4 (9%) and 26 (27%), and Up in 6 (14%) and 25 (26%), respectively. HSV type 1 was found in 2 case samples (5%). Controls were all negative for HSV. Adenovirus was found in 2 NGU samples and none of the controls. In 20 (46%) NGU cases no aetiology was found. Conclusion G. vaginalis, BVAB-2, Eggerthella, L. amnionii, A. vaginae, S. sanguinegens and Prevotella,but not Megasphaeratype 1, had an increasing bacterial load with increasing total Ureaplasma sp. load in male urine regardless of NGU status or co-infections with known NGU pathogens. None of the BV bacteria were associated with NGU. Correlation between total Ureaplasma sp. and BV bacterial load in all samples: Abstract P1.029 Table BV bacteria Spearman-Correlation p-value G. vaginalis 0.59 < 0.0001 BVAB-2 0.28 0.0008 Eggerthella 0.49 < 0.0001 Megasphaera -0.05 0.57 L. amnionii 0.31 0.0003 A. vaginae 0.36 < 0.0001 S. sanguinegens 0.27 < 0.0001 Prevotella sp. 0.14 0.0003

Background Little is known about the presence of bacterial vaginosis (BV) associated bacteria in men, but male partners of women with BV have been reported to have a high risk of urethritis. We aimed to examine the role of BV associated bacteria in urine specimens from men with and without non-gonococcal urethritis (NGU). Methods First-pass urines were collected from 44 men with symptomatic NGU (≥ 5 PMNL/hpf) and 97 asymptomatic men without NGU (< 5 PMNL/hpf). Samples were tested for Chlamydia trachomatis(Ct), Mycoplasma genitalium(Mg), Ureaplasma urealyticum (Uu), U. parvum(Up), HSV 1 and 2, and adenovirus by PCR. Quantitative PCRs were performed to detect Gardnerella vaginalis, BVAB 2, Eggerthella-like uncultured bacterium, Megasphaeratype 1 , Leptotrichia amnionii, Atopobium vaginae, Sneathia sanguinegens, and Prevotella sp. Results Ct was detected in 9 (21%) cases with NGU and 1 (1%) control without NGU. Mg was detected in 10 cases (23%) and none of the controls. Corresponding figures were for Uu 4 (9%) and 26 (27%), and Up in 6 (14%) and 25 (26%), respectively. HSV type 1 was found in 2 case samples (5%). Controls were all negative for HSV. Adenovirus was found in 2 NGU samples and none of the controls. In 20 (46%) NGU cases no aetiology was found. Conclusion G. vaginalis, BVAB-2, Eggerthella, L. amnionii, A. vaginae, S. sanguinegens and Prevotella,but not Megasphaeratype 1, had an increasing bacterial load with increasing total Ureaplasma sp. load in male urine regardless of NGU status or co-infections with known NGU pathogens. None of the BV bacteria were associated with NGU. Correlation between total Ureaplasma sp. and BV bacterial load in all samples:

In the acute phase of leptospirosis, the diagnosis can be established with high sensitivity by testing blood and urine samples with polymerase chain reaction (PCR). However, only few real-time PCR assays have been validated for diagnostic use. The diagnostic accuracy of a novel TaqMan® PCR (LipL32 real-time PCR) targeting the lipl32 gene (or hap-1) and a previously described TaqMan® PCR (16S real-time PCR) targeting the rrs gene coding for 16S rRNA was evaluated when applied to both urine and blood specimens from humans suspected of leptospirosis. Applied to at least two blood cultures LipL32 real-time PCR had a sensitivity of 86%, and a specificity of 100%; and 16S real-time PCR had a sensitivity of 100%, and a specificity of 97%. Applied to urine samples, patients that were positive by the reference methods were also positive by both real-time PCR assays (n=4). For LipL32 real-time PCR the specificity was 100%, while for 16S real-time PCR it was only 91.5% due to unexpected cross-reactions with other bacteria. The analytical sensitivity was close to the theoretical limit-of-detection for both assays detecting all described human pathogenic species. We report a specific real-time PCR assay for detection of Leptospira, i.e., LipL32 real-time PCR that has been validated for diagnostic application in both urine and blood specimens from humans. We further show that a previously described 16S real-time PCR no longer can be recommended for diagnostic use due to a low specificity.

Background Non-gonococcal urethritis (NGU) is a common sexually transmitted disease in men but in 30–50% of NGU cases, no known organism is found. We have used 454 high throughput sequencing to analyse the micro flora in urine samples from cases of urethritis and from controls. Methods Urine samples from men, 10 patients with urethritis and >5 PMNL/hpf and 10 healthy controls with <5 PMNL/hpf, were collected. All samples were tested for Neisseria gonorrhoeae , Chlamydia trachomatis , Mycoplasma genitalium , Ureaplasma urealyticum , U parvum , Trichomonas vaginalis , Herpes Simplex Virus type 1 and 2 and Adenovirus using specific PCR assays. The V3 and V4 regions of the 16S rRNA gene were PCR amplified, tagged and sequenced using the Titanium kit and GS FLX pyrose-quencing system (Roche) according to manufacturer's instructions. Sequences were analysed using the RDP Pyrosequencing Pipeline and CLC Genomics Workbench. Results From each of the 20 samples, 8150 quality filtered sequences were randomly selected. Sequences were assigned to the genus level using the RDP Classifier. A total of 172 gen-era were identified, 133 in patients and 104 in controls. The median number of genera was 35.5 (19–49) in patients with urethritis compared to 25 (16–43) in the controls. No single genus was present in all samples. Members of the genera Pseudomonas and Sphingomonas were present in all the controls and in most of the patient samples. Also, representatives of the genera Brevundimonas, Micrococcus, Bradyrhizobium and Chry-seobacterium were present in 15–18 of the samples. Of the 172 genera, 77 were only found in a single sample and 33 were found in two different samples. Rarefaction analysis at the 5 % level (comparable to genus) suggested that all the controls and most urethritis samples harboured each between 31 and 125 different groups. Four of the urethritis samples apparently contained between 150 and 300 different groups. Conclusions Urine, even from healthy men, contains a very diverse micro-flora. Though not statistically significant, the total and median number of genera was found higher in patients with urethritis than in controls. Several widespread genera are likely to represent commensals and bacteria present in the environment.

Background The aetiology of non-gonococcal urethritis (NGU) is unexplained in 30–50% of cases. The role of ureaplasmas is not clear. We detected Ureaplasma urealyticum (Uu) and U parvum (Up) by quantitative PCR in the urine of men with and without NGU to show a possible association with urethritis. Methods Urine samples from 158 male STD-clinic attendees with symptomatic NGU (>5 PMNL/hpf) and 77 asymptomatic men without NGU (<5 PMNL/hpf) were collected. The patient's age and number of partners within the previous 6 months were recorded. All samples were tested for Neisseria gonorrhoeae (Ng), Chlamydia trachomatis (Ct), Mycoplasma genitalium (Mg), Uu, Up, Trichomonas vaginalis (Tv), herpes simplex virus (HSV) 1 and 2, and adenovirus by real-time PCR. Results Ct and Mg were found in 22 and 30% of NGU, respectively, and were associated with NGU (p<0.0001 both). Three had dual Ct and Mg infection. Uu was detected in 13% of NGU cases and 12% of controls (p>0.99). The median Uu bacterial DNA load was higher in men with NGU than in men without (223 genome equivalents (geq) and 10 geq, respectively; p=0.002). Using ROC-curve analysis to determine the optimal cut-off, patients with >53 geq were more likely to have urethritis (p=0.02). In men with NGU of unknown aetiology, there was no difference in the rate of Uu detection when compared to controls (p=0.26). The corresponding median Uu bacterial DNA load were significantly higher in this group than in controls (p=0.01), and using a cut-off of >53 geq, men with NGU of unknown aetiology were more likely to harbour Uu (14%) than were men from the control group (1%), (p=0.005). Up was detected in 14% of NGU cases and 19% of controls (p=0.34). There was no difference in the detection rate of Up or in the median Up bacterial load in any of the groups. HSV-1 was detected in 3% of cases and 1% of controls. HSV-2 was found in 2% of NGU cases. All urine samples were negative for adenovirus. Cases and controls had similar median number of partners within 6 months (2 partners) and age (28 and 29 years, respectively). Conclusion The bacterial load of U urealyticum in men with NGU and in men with NGU of unknown aetiology was higher than in men without NGU, and the presence of >53 geq of Uu was associated with urethritis in both groups. In accordance with other studies, U parvum was not associated with urethritis. (Preliminary results were presented at the ASM general meeting, San Diego 2010).