Pneumocystis pneumonia (PCP) is a serious fungal lung infection caused by Pneumocystis jirovecii in immunosuppressed individuals. The lack of viable in vitro/ex vivo PCP models has greatly hindered the progress in studying the biology of these fungi, the host/pathogen interactions, as well as antifungal susceptibility testing. In this study, we show the utility of precision-cut lung slices (PCLS) to support the survival of Pneumocystis murina in vitro . We cultured PCLS tissue derived from wild type and immunocompromised mice with a P. murina inoculum in submerged or air-liquid interface models for up to 14 days. We isolated total RNA from the cultured lung tissues at days 0, 3, 7, 10, and 14 and analyzed for the expression of host lung genes and P. murina genes ( Gsc1 for asci and Sp for trophs) by real-time quantitative PCR. When cultured in media alone, P. murina died gradually within a few days. However, when cultured on PCLS, both the troph and ascus forms survived throughout the incubation period of 2 weeks. Moreover, immunohistochemistry staining of P. murina inoculated PCLS sections using polyclonal anti- Pneumocystis sera and showed evidence of fungal aggregation and possible biofilm formation. Additionally, in vitro (PCLS) antibiotic susceptibility testing using commonly used antifungal drugs confirmed successful targeting of the troph and ascus forms by trimethoprim sulfamethoxazole and the ascus form by echinocandins. IMPORTANCE: Our study reveals the potential of precision-cut lung slices as an ex vivo platform to study the growth/survival of Pneumocystis spp. that can facilitate the development of new anti-fungal drugs.

Introduction: The objective of this narrative review is to examine the health risks associated with informal mining in Sub-Saharan Africa (SSA), with a particular focus on HIV prevention. It aims to review existing interventions targeting this population and identify challenges and opportunities for improvement. Methods: A comprehensive literature review was conducted using databases such as Google Scholar, PubMed, ScienceDirect, and Cochrane Library. Studies and reports published between 2000 and 2023 that focused on occupational health risks and HIV prevention interventions in the informal mining sector of SSA were included in the analysis. Results: Informal mining in SSA presents significant health risks to workers, including exposure to hazardous substances, poor working conditions, and limited access to healthcare. These factors, combined with the high prevalence of HIV/AIDS in the region, make informal miners particularly vulnerable to infection. The review identified several key themes related to occupational health risks, such as poor environmental and sanitation conditions, increased vulnerability due to factors like poverty and lack of education, and limited access to healthcare services. A variety of interventions have been implemented to address HIV prevention among informal miners in SSA. These include behavioral strategies like peer education and social marketing campaigns, as well as biomedical interventions such as counseling, testing, and pre-exposure prophylaxis (PrEP). However, challenges persist in delivering effective HIV prevention services to this population. These challenges include restricted access to healthcare, the high mobility of miners, limited resources, and stigma associated with HIV. Conclusions: The findings of this review highlight the urgent need for integrated health services and tailored interventions that address the specific issues faced by informal miners in SSA. Community-based and culturally sensitive programs, developed in collaboration with mining communities, are essential for effective HIV prevention. Future research should evaluate the long-term impact of interventions and explore their scalability. To improve intervention effectiveness and sustainability, stakeholders should focus on community engagement, strengthening health systems, and addressing structural barriers. A multi-sectoral approach is necessary to tackle the broader determinants of health in mining communities.