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We aimed to determine the household distribution and viability of Chlamydia trachomatis (Ct) from the eyes, face, and hands during the initial two visits of a year-long fortnightly cohort study in geographically defined adjacent households. We enrolled 298 individuals from 68 neighbouring households in Shashemene Woreda, Oromia, Ethiopia. All individuals above 2 years of age residing in these households were examined for signs of trachoma. Swab samples were taken from the conjunctiva, faces, and hands and analysed for the presence and viability of Ct. Ct viability was determined using reverse transcription (RT) PCR. At the initial visit, out of 298 individuals, 133 (44.5%) were children aged 2-9 years. Among these children, 27/133 (20.3%) had trachomatous inflammation-follicular (TF), while 8/133 (6.0%) had trachomatous inflammation-intense (TI). Ct (omcB or pORF2) was detected in 16/133 (12.0%) eye swabs, 14/105 (13.5%) face swabs, and 11/105 (10.5%) hand swabs from children aged 2-9 years. Among these children at visit one, 12/14 (85.7%) with Ct on faces and 9/11 (81.8%) with Ct on hands also had detectable ocular Ct. The severity of the disease worsened from the first visit to the second, and no participants showed clearance of the disease within the two-week period. Ct infection was associated with TF (P = 0.002) and TI (P = 0.060). At visit one, among children aged 2-9 years, viable Ct was detected in 12/16 (75.0%) ocular, 6/14 (42.9%) face, and 4/11 (36.4%) hand swab samples. All viable Ct detected on the faces and hands were identified from individuals with viable ocular infections. Among caregivers whose child tested positive for Ct on their hands, 3 caregivers also had Ct on their hands, accounting for 20% (3 out of 15). Additionally, among caregivers whose child tested positive for Ct on their faces, 2 caregivers had Ct on their faces, which accounts for 14.3% (2 out of 14). In two participants, we detected Ct on the hands of ocular-negative children at the initial visit and later detected ocular Ct at the second visit. Using RT-qPCR assay to detect Ct omp2 mRNA to define viability offers a new, informative perspective of trachoma transmission in this community in Ethiopia. The presence of viable Ct on the faces and hands of individuals living in households with people with current ocular Ct infection supports the hypothesis that hands and faces are important routes for transmission of trachoma. This highlights the importance of targeted interventions to address these sites of Ct carriage to help interrupt transmission.

IntroductionTrachoma is caused by the bacterium Chlamydia trachomatis (Ct). The WHO recommends the SAFE strategy for trachoma elimination: Surgery for trichiasis, Antibiotics, Facial cleanliness and Environmental improvement. Multiple rounds of SAFE implementation have proven insufficient to eliminate trachoma in Ethiopia, where over 50% of the global trachoma burden remains. More effective antibiotic treatment schedules and transmission-suppressing approaches are needed. The aim of stronger SAFE is to evaluate the impact of a novel package of interventions to strengthen the A, F and E of SAFE on the prevalence of ocular Ct and trachoma in Oromia, Ethiopia.Methods and analysis68 clusters were randomised in a 1:1:1:1 ratio to one of (1) standard A/standard F&E (standard SAFE), (2) standard A/enhanced F&E, (3) enhanced A/standard F&E or (4) enhanced A/enhanced F&E (stronger SAFE). Enhanced A includes two height-based doses of oral azithromycin (equivalent to 20 mg/kg) given as single doses 2 weeks apart, as mass drug administration, annually. Enhanced F&E includes fly control measures (permethrin-treated headwear and odour-baited traps) and face-washing hygiene behaviour change implemented at household level in selected communities. The interventions will be implemented and reinforced over 3 years.The primary outcome is the prevalence of ocular Ct by quantitative PCR in children aged 1–9 years at 36 months. A key secondary outcome is the prevalence of active (inflammatory) trachoma in the same children, assessed by validated trachoma graders and conjunctival photography. Laboratory technicians and photo-graders are masked to treatment allocation. Other important secondary analyses include process evaluations, assessment of behaviour change, fly indicators, adherence and coverage of interventions and a cost analysis.Ethics and disseminationStudy protocols have been approved by the National Research Ethics Review Committee of the Ethiopian Ministry of Science and Higher Education and the London School of Hygiene & Tropical Medicine Ethics Committee. An independent data safety and monitoring board oversees the trial. Results will be disseminated through peer-reviewed publications, presentations and reports.Trial registration number ISRCTN40760473.