The aim of the present study was to investigate whether addition of the BACTEC™ Mycosis bottle to the standard BACTEC™ aerobic and anaerobic bottles contributed to a higher detection rate and a faster time to detection (TTD) of fungi. This was a retrospective cohort study of all patients with a positive blood culture with Candida species delivered to the Department of Clinical Microbiology, Herlev and Gentofte Hospital, Denmark in the 8-year period 2006 through 2014. The patients had at least one BACTEC™ aerobic and one Mycosis bottle sampled at the same time and at least one of the bottles yielded growth of fungi. Among 184 patients included, 173 were examined using BACTEC™ aerobic, anaerobic and Mycosis bottles. The anaerobic vial generally had the lowest detection rate and the longest TTD. The detection rate of BACTEC™ aerobic plus anaerobic with the BACTEC™ Mycosis bottle was significantly higher than the detection rate of BACTEC™ aerobic plus anaerobic without BACTEC™ Mycosis bottle for all species after 1–5 days, and specially for Candida glabrata at 2, 3, 4 and 5 days. TTD for C. glabrata was significantly shorter for BACTEC™ Mycosis than TTD for BACTEC™ aerobic or anaerobic bottles after ½ to 4 days. When combining “first or only” detection, the BACTEC™ Mycosis bottle had a significantly higher detection as compared to the aerobic bottle. Addition of the BACTEC™ Mycosis bottle to the standard BACTEC™ aerobic and anaerobic bottles significantly contributed to a higher detection rate and a faster TTD of fungemia.

BACKGROUND:Bacterial vaginosis (BV) is an imbalance of the vaginal bacterial microbiota and its aetiology is still unknown. Our aims were to investigate the diagnostic potential of species/genus specific quantitative PCR (qPCR) for bacteria present in swabs and first-void urine (FVU) samples using Nugent's and Claeys' criteria and 454 sequencing of the vaginal microbiome as reference.METHODS:Self-collected swabs, vaginal smears and FVU were obtained from 177 women from Greenland (Study I and III) and physician-collected vaginal swabs and smears were obtained from 163 Swedish women (Study II). BV was diagnosed by Nugent's criteria in Study I and III and by Amsel's criteria in Study II. The vaginal swabs and FVU samples were analysed by qPCR for selected vaginal bacteria in all three studies and for four sexually transmitted infections (STIs) in Study I.RESULTS:Study I: STIs were common in women from Greenland and BV was found in 45% of these women but was not associated with individual STIs. In multivariate logistic analysis, Atopobium vaginae and Prevotella spp. were both independently associated with BV in swabs. BV could be subdivided into clusters dominated by a single or a few species together. Seven vaginal bacteria (A. vaginae, Prevotella spp. Gardnerella vaginalis, Bacterial vaginosis associated bacterium (BVAB) 2, Eggerthella-like bacterium, Leptotrichia amnionii and Megasphaera type 1) had areas under the receiver operating characteristic (ROC) curve > 85% in swabs, suggesting that they were good predictors of BV according to Nugent. Study II: For the majority of species/genera, the kappa values indicated fair to good agreement when their presence was determined by 454 pyrosequencing versus real-time PCR. The same seven vaginal bacteria as found in Study I, had areas under the ROC-curve > 85% in swabs from Swedish women, demonstrating a good diagnostic accuracy for BV according to Amsel. Study III: In a multivariate model, Megasphaera type 1 and Prevotella spp. remained significantly associated with BV in FVU samples. A linear regression analysis showed good agreement between bacterial load from swabs and FVU, but Prevotella spp. could be detected in high numbers in a few FVU samples without being present in swabs. After applying ROC curve analysis, the same seven vaginal bacteria as previously mentioned showed good prediction for BV according to Nugent in FVU. BV could be detected with comparable sensitivity in FVU and vaginal swabs.CONCLUSION:BV can be diagnosed by molecular methods performed either on swabs or urine but it is important to apply thresholds in order to improve the accuracy of the diagnosis. Furthers it was possible to identify clusters of BV dominated by single or paired bacteria, and these clusters could classify BV into subgroups, providing a more detailed understanding of the condition. Seven vaginal bacteria were highly accurate for BV diagnosis both in swabs and FVU. Finally a good agreement between Nugent and Claeys was found.

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:

Background Bacterial vaginosis (BV) is the most common vaginal infection/disorder. BV is characterised by imbalance in the normal vaginal microbiota with a shift towards higher bacterial diversity and increased pH. The aim of the present study was to describe the differences in vaginal microbiota composition in women suffering from BV compared to healthy women, using massive parallel 454 pyrosequencing. Methods 163 vaginal samples were collected from women diagnosed with characteristic BV (n=73), women with intermediate BV (n=11), and from healthy women on their regular check-ups (n=79). DNA from the samples was isolated and the bacterial compositions as well as the relative abundance of these bacteria were analysed using 454 pyrosequencing, with GS Titanium amplicons kit (Roche Inc.), of the hypervariable region V4 on the 16S rRNA gene. Finally, 17 different species-specific PCRs were used to verify the species of bacteria found in the 454 pyrosequencing. Results Extensive imbalance of the vaginal microbiota of women with BV compared to healthy controls was revealed. The dominating taxons of the 73 BV cases were Gardnerella, Atopobium, Prevotella, Lactobacillus, Megasphera and Sneathia, while most of the 79 healthy controls had a microbiota totally dominated by Lactobacillus with the BV associated taxons hardly detectable. Furthermore, the 11 patients with intermediate BV predominantly had a mix of the BV associated taxon Gardnerella as well as Lactobacillus. A few of the healthy controls seemed to have a microbiota changing towards the intermediate microflora. Gardnerella may be the first bacteria to establish in the transition from healthy vaginal flora towards a BV associated flora. Conclusions A clear difference in the composition of the vaginal microbiota between individuals suffering from BV and healthy controls was identified. The present findings are important steps towards the determination of valid potential bacterial markers for BV, are shedding light upon why some women develop BV, as well as show how the microbiota is involved in the development of BV. Knowledge of the composition of the vaginal microbiota is crucial in the development of a BV diagnostic tool and for elucidating appropriate treatment for use in clinical practice.

Background The aetiology of non-gonococcal urethritis (NGU) in men is unknown in 30%–50% of cases. Little is known about the relation of bacterial vaginosis (BV) associated bacteria in men with urethritis of unknown aetiology (UUE). Urethral swabs from men with and without NGU were analysed with PCR for BV associated bacteria to show a possible association with UUE. Methods Urethral swabs from 9 and 19 men with symptomatic and asymptomatic NGU (>5 PMNL/hpf), respectively, and 30 asymptomatic men without NGU were collected. All samples were negative for Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium , Ureaplasma urealyticum and U parvum with specific PCR assays. Quantitative real-time PCR with TaqMan based assays were performed to detect Atopobium vaginae (Av), Sneathia sanguinegens (Ss), Leptotrichia amnionii (La) and Gardnerella vaginalis (Gv), and with SYBR green assays for BVAB 1, 2 and 3, and Megasphaera phylotype 1 (M1). Results Gv was detected in 93% of cases with UUE and in 37% of controls (p<0.0001). There was no difference in organism load. In the 28 NGU cases Av, Ss and La were found in 3, 2 and 1 samples, respectively, and in 6, 2 and 1 of the control samples, respectively. The median corresponding organism loads were 14, 95 and 51 for the NGU cases and 16, 10 566 and 353 for the controls. All samples were negative for BVAB 1, 2 and 3 and M1, except one control with 10 genome copies of BVAB 1. Conclusions Gardnerella vaginalis was associated with male urethritis in this study, especially in men with asymptomatic urethritis, while BVAB 1, 2 and 3, Megasphaera phylotype 1, Atopobium vaginae , Sneathia sanguinegens , and Leptotrichia amnionii were not.

A 41-year old multisectio patient got a life-threatening postoperative infection with Gardnerella vaginalis and Peptostreptococcus spp. Perioperative treatment with cefuroxime and metronidazol is recommended.