Molecular investigation of Treponema pallidum strains associated with ocular syphilis in the United States, 2016-2020.

Ocular syphilis is a serious complication of Treponema pallidum infection that can occur at any stage of syphilis and affect any eye structure. It remains unknown if certain T. pallidum strains are associated with ocular infections; therefore, we performed genotyping and whole genome sequencing (WGS) to characterize strains from patients with ocular syphilis. Seventy-five ocular or non-ocular specimens from 55 ocular syphilis patients in 14 states within the United States were collected between February 2016 and November 2020. Sufficient T. pallidum DNA was available from nine patients for genotyping and three for WGS. Genotyping was done using the augmented Centers for Disease Control and Prevention typing scheme, and WGS was performed on Illumina platforms. Multilocus sequence typing allelic profiles were predicted from whole genome sequence data. T. pallidum DNA was detected in various specimens from 17 (30.9%) of the 55 patients, and typing was done on samples from 9 patients. Four complete strain types (14d10/g, 14b9/g, 14d9/g, and 14e9/f) and five partial types were identified. WGS was successful on samples from three patients and all three strains belonged to the SS14 clade of T. pallidum. Our data reveal that multiple strain types are associated with ocular manifestations of syphilis. While genotyping and WGS were challenging due to low amounts of T. pallidum DNA in specimens, we successfully performed WGS on cerebrospinal fluid, vitreous fluid, and whole blood.IMPORTANCESyphilis is caused by the spirochete Treponema pallidum. Total syphilis rates have increased significantly over the past two decades in the United States, and the disease remains a public health concern. In addition, ocular syphilis cases has also been on the rise, coinciding with the overall increase in syphilis rates. We conducted a molecular investigation utilizing traditional genotyping and whole genome sequencing over a 5-year period to ascertain if specific T. pallidum strains are associated with ocular syphilis. Genotyping and phylogenetic analysis show that multiple T. pallidum strain types are associated with ocular syphilis in the United States.

'Protecting your body and loving yourself': strategies Black male caregivers use to protect Black girls sexuality in the USA.

Black girls in the USA face disproportionate rates of sexually transmitted infections (STIs), HIV, and sexual violence, prompting research into the sociocultural factors such as adultification, race and gender discrimination, and stereotyped messaging, impacting Black girls' sexual development. While existing literature finds that Black female caregivers utilise protective strategies to mitigate potential harm to Black girls, little is known about the role of Black male caregivers. This qualitative study employed the Becoming a Sexual Black Woman framework and one-on-one interviews to examine how 30 Black male caregivers of girls conceptualised parental protection and elated strategies. The thematic analysis revealed key themes of: Being a Provider, Being a Present Role Model, Protecting Black Girls' Bodies, Limiting Exposure to Stereotyped Messages, and Providing Emotional Protection. Black male caregivers navigated traditional gendered expectations and limited resources while utilising parental strategies that reflected patriarchal, sometimes misogynistic, and non-patriarchal ideals and norms. The study highlights the attitudes and beliefs of Black male caregivers regarding protecting Black girls from sexual violence and other related adverse outcomes. The findings offer insights for family-based interventions and programmes that empower Black girls to support their sexual development and health in the USA.

Application of ADAPT-ITT: adapting an evidence-based HIV/STI mother-daughter prevention intervention for Black male caregivers and girls.

BACKGROUND: Black girls are disproportionately impacted by HIV and sexually transmitted infections (STIs), underscoring the urgent need for innovative strategies to enhance the adoption and maintenance of HIV/STI prevention efforts. Historically, Black male caregivers have been left out of girls' programming, and little guidance exists to inform intervention development for Black girls and their male caregivers. Engaging Black male caregivers in Black girls' sexual and reproductive health may reduce sexual risk-taking and improve the sustainability of preventative behaviors. OBJECTIVE: This paper describes the formative phases, processes, and methods used to adapt an evidence-based mother-daughter sexual and reproductive health intervention for Black girls 9-18 years old and their male caregivers. METHODS: We used the ADAPT-ITT model to tailor IMARA for Black girls and their male caregivers. Diverse qualitative methods (interviews, focus groups, and theater testing) were used throughout the adaption process. RESULTS: Findings support using the ADAPT-ITT model to tailor an evidence-based HIV/STI intervention for Black girls and their Black male caregivers. Findings highlight the importance of community engagement and the use of qualitative methods to demonstrate the acceptability and feasibility of the adapted intervention. Key lessons learned are reviewed. CONCLUSIONS: Adapting evidence-based interventions to incorporate Black girls and their Black male caregivers should be driven by a relevant theoretical framework that aligns with the target population(s). Adapting the intervention in partnership with the community has been shown to improve acceptability and feasibility as it is responsive to community needs. Using a systematic process like the ADAPT-ITT model will ensure that the new program is ready for efficacy trials.

Selective Whole-Genome Amplification as a Tool to Enrich Specimens with Low Treponema pallidum Genomic DNA Copies for Whole-Genome Sequencing.

Downstream next-generation sequencing (NGS) of the syphilis spirochete Treponema pallidum subspecies pallidum (T. pallidum) is hindered by low bacterial loads and the overwhelming presence of background metagenomic DNA in clinical specimens. In this study, we investigated selective whole-genome amplification (SWGA) utilizing multiple displacement amplification (MDA) in conjunction with custom oligonucleotides with an increased specificity for the T. pallidum genome and the capture and removal of 5'-C-phosphate-G-3' (CpG) methylated host DNA using the NEBNext Microbiome DNA enrichment kit followed by MDA with the REPLI-g single cell kit as enrichment methods to improve the yields of T. pallidum DNA in isolates and lesion specimens from syphilis patients. Sequencing was performed using the Illumina MiSeq v2 500 cycle or NovaSeq 6000 SP platform. These two enrichment methods led to 93 to 98% genome coverage at 5 reads/site in 5 clinical specimens from the United States and rabbit-propagated isolates, containing >14 T. pallidum genomic copies/µL of sample for SWGA and >129 genomic copies/µL for CpG methylation capture with MDA. Variant analysis using sequencing data derived from SWGA-enriched specimens showed that all 5 clinical strains had the A2058G mutation associated with azithromycin resistance. SWGA is a robust method that allows direct whole-genome sequencing (WGS) of specimens containing very low numbers of T. pallidum, which has been challenging until now. IMPORTANCE Syphilis is a sexually transmitted, disseminated acute and chronic infection caused by the bacterial pathogen Treponema pallidum subspecies pallidum. Primary syphilis typically presents as single or multiple mucocutaneous lesions and, if left untreated, can progress through multiple stages with various clinical manifestations. Molecular studies often rely on direct amplification of DNA sequences from clinical specimens; however, this can be impacted by inadequate samples due to disease progression or timing of patients seeking clinical care. While genotyping has provided important data on circulating strains over the past 2 decades, WGS data are needed to better understand strain diversity, perform evolutionary tracing, and monitor antimicrobial resistance markers. The significance of our research is the development of an SWGA DNA enrichment method that expands the range of clinical specimens that can be directly sequenced to include samples with low numbers of T. pallidum.

The Impact of COVID-19 Among Black Girls: A Social-Ecological Perspective.

OBJECTIVE: To explore the impact of the COVID-19 pandemic on Black adolescent girls' physical, psychological, and sexual development. METHODS: Semistructured interviews were conducted with 25 girls (9-18 years old) primarily across the Midwestern United States. Interviews were conducted until theoretical saturation was reached. We conducted a conventional content analysis by reviewing transcripts, coding data, and developing themes. RESULTS: Utilizing the social-ecological model findings revealed themes related to the individual-, interpersonal-, institutional-, and community-level changes that Black girls experienced during the COVID-19 pandemic. CONCLUSIONS: Black adolescent girls need opportunities to discuss the changes, receive relevant resources, and have spaces to support their physical, psychological, and sexual development. Systems such as families, schools, and clinics should implement programming to facilitate discussions around the impact of COVID-19 on Black girls.

Successful isolation of Treponema pallidum strains from patients' cryopreserved ulcer exudate using the rabbit model.

Clinical isolates of Treponema pallidum subspecies pallidum (T. pallidum) would facilitate study of prevalent strains. We describe the first successful rabbit propagation of T. pallidum from cryopreserved ulcer specimens. Fresh ulcer exudates were collected and cryopreserved with consent from syphilis-diagnosed patients (N = 8). Each of eight age-matched adult male rabbits were later inoculated with a thawed specimen, with two rabbits receiving 1.3 ml intratesticularly (IT), and six receiving 0.6 ml intravenously (IV) and IT. Monitoring of serology, blood PCR and orchitis showed that T. pallidum grew in 2/8 rabbits that were inoculated IV and IT with either a penile primary lesion specimen (CDC-SF003) or a perianal secondary lesion specimen (CDC-SF007). Rabbit CDC-SF003 was seroreactive by T. pallidum Particle Agglutination (TP-PA) and Rapid Plasma Reagin (RPR) testing, PCR+, and showed orchitis by week 6. Euthanasia was performed in week 7, with treponemal growth in the testes confirmed and quantified by qPCR and darkfield microscopy (DF). Serial passage of the extract in a second age-matched rabbit also yielded treponemes. Similarly, rabbit CDC-SF007 showed negligible orchitis, but was seroreactive and PCR+ by week 4 and euthanized in week 6 to yield T. pallidum, which was further propagated by second passage. Using the 4-component molecular typing system for syphilis, 3 propagated strains (CDC-SF003, CDC-SF007, CDC-SF008) were typed as 14d9f, 14d9g, and 14d10c, respectively. All 3 isolates including strain CDC-SF011, which was not successfully propagated, had the A2058G mutation associated with azithromycin resistance. Our results show that immediate cryopreservation of syphilitic ulcer exudate can maintain T. pallidum viability for rabbit propagation.