The magnitude of the 2022 multi-country monkeypox virus (MPXV) outbreak has surpassed any preceding outbreak. It is unclear whether asymptomatic or otherwise undiagnosed infections are fuelling this epidemic. In this study, we aimed to assess whether undiagnosed infections occurred among men attending a Belgian sexual health clinic in May 2022. We retrospectively screened 224 samples collected for gonorrhea and chlamydia testing using an MPXV PCR assay and identified MPXV-DNA-positive samples from four men. At the time of sampling, one man had a painful rash, and three men had reported no symptoms. Upon clinical examination 21–37 days later, these three men were free of clinical signs, and they reported not having experienced any symptoms. Serology confirmed MPXV exposure in all three men, and MPXV was cultured from two cases. These findings show that certain cases of monkeypox remain undiagnosed and suggest that testing and quarantining of individuals reporting symptoms may not suffice to contain the outbreak.

Objectives Bacterial vaginosis (BV) is characterised by a change in the microbial composition of the vagina. The BV-associated organisms outnumber the health-associated Lactobacillus species and form a polymicrobial biofilm on the vaginal epithelium, possibly explaining the difficulties with antibiotic treatment. A better understanding of vaginal biofilm with emphasis on Atopobium vaginae and Gardnerella vaginalis may contribute to a better diagnosis and treatment of BV. Methods To this purpose, we evaluated the association between the presence of both bacteria by fluorescence in situ hybridisation (FISH) and BV by Nugent scoring in 463 vaginal slides of 120 participants participating in a clinical trial in Rwanda. Results A bacterial biofilm was detected in half of the samples using a universal bacterial probe. The biofilm contained A. vaginae in 54.1% and G. vaginalis in 82.0% of the samples. A. vaginae was accompanied by G. vaginalis in 99.5% of samples. The odds of having a Nugent score above 4 were increased for samples with dispersed G. vaginalis and/or A. vaginae present (OR 4.5; CI 2 to 10.3). The probability of having a high Nugent score was even higher when a combination of adherent G. vaginalis and dispersed A. vaginae was visualised (OR 75.6; CI 13.3 to 429.5) and highest when both bacteria were part of the biofilm (OR 119; CI 39.9 to 360.8). Conclusions Our study, although not comprehensive at studying the polymicrobial biofilm in BV, provided a strong indication towards the importance of A. vaginae and the symbiosis of A. vaginae and G. vaginalis in this biofilm. Trial registration number NCT01796613.

Screening for STIs in men who have sex with men receiving HIV pre-exposure prophylaxis resulted in high consumption of macrolides, extended spectrum cephalosporins, fluoroquinolones and tetracyclines. The consumption of macrolides was 52 times as high as that reported for community-level consumption in certain European countries.

Hormonal contraception has been associated with a reduced risk of vaginal dysbiosis, which in turn has been associated with reduced prevalence of sexually transmitted infections (STIs), including HIV. Vaginal rings are used or developed as delivery systems for contraceptive hormones and antimicrobial drugs for STI and HIV prevention or treatment. We hypothesized that a contraceptive vaginal ring (CVR) containing oestrogen enhances a lactobacilli-dominated vaginal microbial community despite biomass accumulation on the CVR’s surface. We enrolled 120 women for 12 weeks in an open-label NuvaRing^® study at Rinda Ubuzima, Kigali, Rwanda. Vaginal and ring microbiota were assessed at baseline and each ring removal visit by Gram stain Nugent scoring (vaginal only), quantitative PCR for Lactobacillus species, Gardnerella vaginalis and Atopobium vaginae, and fluorescent in situ hybridization to visualize cell-adherent bacteria. Ring biomass was measured by crystal violet staining. Bacterial vaginosis (BV) prevalence was 48% at baseline. The mean Nugent score decreased significantly with ring use. The presence and mean log₁₀ concentrations of Lactobacillus species in vaginal secretions increased significantly whereas those of G. vaginalis and presence of A. vaginae decreased significantly. Biomass accumulated on the CVRs with a species composition mirroring the vaginal microbiota. This ring biomass composition and optical density after crystal violet staining did not change significantly over time. NuvaRing^® promoted lactobacilli-dominated vaginal microbial communities in a population with high baseline BV prevalence despite the fact that biomass accumulated on the rings.

Background Monkeypox is transmitted by close contact with symptomatic cases, and those infected are assumed to be uniformly symptomatic. Evidence of subclinical monkeypox infection is limited to a few immunological studies which found evidence of immunity against orthopoxviruses in asymptomatic individuals who were exposed to monkeypox cases. We aimed to assess whether asymptomatic infections occurred among individuals who underwent sexually transmitted infection (STI) screening in a large Belgian STI clinic around the start of the 2022 monkeypox epidemic in Belgium. Methods Anorectal and oropharyngeal swabs collected for gonorrhoea/chlamydia screening from May 1 until May 31, 2022 were retrospectively tested by a monkeypox-specific PCR. Cases with a positive PCR result were recalled to the clinic for case investigation, repeat testing and contact tracing. Findings In stored samples from 224 men, we identified three cases with a positive anorectal monkeypox PCR. All three men denied having had any symptoms in the weeks before and after the sample was taken. None of them reported exposure to a diagnosed monkeypox case, nor did any of their contacts develop clinical monkeypox. Follow-up samples were taken 21 to 37 days after the initial sample, by which time the monkeypox-specific PCR was negative, likely as a consequence of spontaneous clearance of the infection. Interpretation The existence of asymptomatic monkeypox infection indicates that the virus might be transmitted to close contacts in the absence of symptoms. Our findings suggest that identification and isolation of symptomatic individuals may not suffice to contain the outbreak. Funding Institutional funding Evidence before this study Similar to smallpox, monkeypox is transmitted through close contact with symptomatic cases, and 100% of those infected are assumed to develop symptoms. These features imply that an outbreak in the general population tends towards extinction with relatively minor hygienic measures, as observed in several outbreaks in endemic regions. If, however, asymptomatic transmission occurs, the outbreak becomes much more difficult to contain. We searched PubMed and Google Scholar for evidence of asymptomatic human monkeypox, using the search terms “monkeypox” AND (“asymptomatic” OR “subclinical”), and included peer-reviewed reports published until June 17, 2022. We identified seven original reports in three different epidemiological settings which reported indirect, immunological evidence of asymptomatic monkeypox infection in a small number of people who were exposed to the virus. We did not find any study that provided direct evidence of the virus in asymptomatic individuals. Added value of this study By retrospectively screening clinical samples collected for sexually transmitted infection screening in our centre throughout May 2022 with a monkeypox-specific PCR, we found evidence of asymptomatic monkeypox virus infection in three individuals. Implications of all the available evidence The existence of asymptomatic monkeypox infection indicates that the virus may be transmitted in the absence of symptoms. This risk can be further quantified by studying viral dynamics in contacts of symptomatic and asymptomatic monkeypox cases. Our findings suggest that identification and isolation of symptomatic individuals may not suffice to contain the outbreak.

In this study, we characterized all oropharyngeal and anorectal isolates of Neisseria spp. in a cohort of men who have sex with men. This resulted in a panel of pathogenic Neisseria (N. gonorrhoeae [n = 5] and N. meningitidis [n = 5]) and nonpathogenic Neisseria (N. subflava [n = 11], N. mucosa [n = 3] and N. oralis [n = 2]). A high proportion of strains in this panel were resistant to azithromycin (18/26) and ceftriaxone (3/26). Whole genome sequencing (WGS) of these strains identified numerous mutations that are known to confer reduced susceptibility to azithromycin and ceftriaxone in N. gonorrhoeae. The presence or absence of these known mutations did not explain the high level resistance to azithromycin (>256 mg/L) in the nonpathogenic isolates (8/16). After screening for antimicrobial resistance (AMR) genes, we found a ribosomal protection protein, Msr(D), in these highly azithromycin resistant nonpathogenic strains. The complete integration site originated from Streptococcus pneumoniae and is associated with high level resistance to azithromycin in many other bacterial species. This novel AMR resistance mechanism to azithromycin in nonpathogenic Neisseria could be a public health concern if it were to be transmitted to pathogenic Neisseria. This study demonstrates the utility of WGS-based surveillance of nonpathogenic Neisseria.