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Background: Accuracy of molecular tools for the identification of parasites that cause human cutaneous leishmaniasis (CL) could largely depend on the sampling method. Non-invasive or less-invasive sampling methods such as filter paper imprints and cotton swabs are preferred over punch biopsies and lancet scrapings for detection methods of Leishmania based on polymerase chain reaction (PCR) because they are painless, simple, and inexpensive, and of benefit to military and civilian patients to ensure timely treatment. However, different types of samples can generate false negatives and there is a clear need to demonstrate which sample is more proper for molecular assays. Methodology: Here, we compared the sensitivity of molecular identification of different Leishmania (Viannia) species from Peru, using three types of sampling: punch biopsy, filter paper imprint and lancet scraping. Different composite reference standards and latent class models allowed to evaluate the accuracy of the molecular tools. Additionally, a quantitative PCR assessed variations in the results and parasite load in each type of sample. Principal findings: Different composite reference standards and latent class models determined higher sensitivity when lancet scrapings were used for sampling in the identification and determination of Leishmania (Viannia) species through PCR-based assays. This was consistent for genus identification through kinetoplastid DNA-PCR and for the determination of species using FRET probes-based Nested Real-Time PCR. Lack of species identification in some samples correlated with the low intensity of the PCR electrophoretic band, which reflects the low parasite load in samples. Conclusions: The type of clinical sample can directly influence the detection and identification of Leishmania (Viannia) species. Here, we demonstrated that lancet scraping samples consistently allowed the identification of more leishmaniasis cases compared to filter paper imprints or biopsies. This procedure is inexpensive, painless, and easy to implement at the point of care and avoids the need for anesthesia, surgery, and hospitalization and therefore could be used in resource limited settings for both military and civilian populations. Author summary: Human cutaneous leishmaniasis affects low-income populations living in places far from health services. The early sampling and detection of the parasite are necessary for timely treatment, however there are no uniform sampling criteria, thus the sensitivity of molecular tests may vary due to various factors such as the type of sample, the time of the disease and the parasite load in the lesion. In this study, we compared the performance of three sampling methods for molecular identification of the genus and Leishmania (Viannia) species in Peru. Several analytical methods, including composite reference standards and latent class models, suggested that lancet scraping might be the best approach for parasite genus detection by kDNA-PCR and for parasite species determination by FRET probes-based Nested Real-Time PCR. [ABSTRACT FROM AUTHOR]

Leishmaniases are a group of neglected vector-borne infectious diseases that are among the six priority endemic diseases worldwide. Visceral leishmaniasis (VL) is the most severe clinical manifestation, characterized by systemic and chronic visceral involvement and high mortality in immunosuppressed and untreated patients. VL can be complicated into post-kala-azar dermal leishmaniasis (PKDL), and when dermatologic disorders occur simultaneously with active VL, an intermediate clinical form called para-kala-azar dermal leishmaniasis (para-KDL) occurs. This clinical form is of great epidemiological relevance, as humans act as a source of infection for vectorial transmission. In the Americas, Brazil is among the seven countries responsible for more than 90% of VL cases, though reports of PKDL and para-KDL are rare. This paper presents three cases of VL-HIV co-infection with Leishmania-containing skin lesions resembling para-kala-azar dermal leishmaniasis. The cases were investigated by the team from the Infectious Diseases Department of University Hospital (HUMAP/UFMS) in Mato Grosso do Sul, Brazil. The three patients exhibited skin lesions where amastigote forms of L. (L.) infantum were identified. All cases exhibited similar clinical manifestations of para-KDL, including fever, hepatosplenomegaly, pancytopenia, and disseminated skin lesions. The study described the prevalence of comorbidities, the incidence of VL relapse, and the therapeutic regimen in relation to the outcomes. The study underscores the importance of follow-up and secondary prophylaxis in patients with VL, which are essential for the efficacy of the treatment. Furthermore, the study provides insight into the potential epidemiological profile of para-KDL cases in Brazil, which contributes to the development of more efficient clinical management strategies for patients. Author summary: Visceral leishmaniasis (VL) is a neglected vector-borne infectious disease with a systemic and chronic involvement that is lethal when untreated. Para-kala-azar dermal leishmaniasis (para-KDL) is a complication of VL, in which the patient presents with dermatological and visceral manifestations simultaneously. Para-KDL cases are of significant epidemiological importance, as humans act as a source of infection in parasite transmission. In the Americas, Brazil accounts for over 90% of VL cases. Nevertheless, reports of para-KDL remain scarce. This paper presents three cases of VL-HIV co-infection with Leishmania-containing skin lesions resembling para-kala-azar dermal leishmaniasis. The three cases exhibit similarities in terms of signs and symptoms, diagnostic criteria for AIDS, and the presence of active VL with skin lesions. The study contributes to the understanding of the epidemiological profile of para-PKDL cases in Brazil. It highlights the significance of multidisciplinary follow-up and secondary prophylaxis for patients with VL and emphasizes the need for more effective clinical management strategies for these patients. [ABSTRACT FROM AUTHOR]

Background: Human African trypanosomiasis (HAT) is a neglected tropical disease that usually occurs in rural areas in sub-Saharan Africa. It caused devastating epidemics during the 20th century. Sustained, coordinated efforts by different stakeholders working with national sleeping sickness control programmes (NSSCPs) succeeded in controlling the disease and reducing the number of cases to historically low levels. In 2012, WHO targeted the elimination of the disease as a public health problem by 2020. This goal has been reached and a new ambitious target was stated in the WHO road map for NTDs 2021–2030 and endorsed by the 73rd World Health Assembly: the elimination of gambiense HAT transmission (i.e. reducing the number of reported cases to zero). The interruption of transmission was not considered as an achievable goal for rhodesiense HAT, as it would require vast veterinary interventions rather than actions at the public health level. Methodology/principal findings: Data reported to WHO by NSSCPs were harmonized, verified, georeferenced and included in the atlas of HAT. A total of 802 cases were reported in 2021 and 837 in 2022. This is below the target for elimination as a public health problem at the global level (< 2000 HAT cases/year); 94% of the cases were caused by infection with T. b. gambiense. The areas reporting ≥ 1 HAT case/10 000 inhabitants/year in 2018–2022 cover a surface of 73 134 km2, with only 3013 km2 at very high or high risk. This represents a reduction of 90% from the baseline figure for 2000–2004, the target set for the elimination of HAT as a public health problem. For the surveillance of the disease, 4.5 million people were screened for gambiense HAT with serological tests in 2021–2022, 3.6 million through active screening and 0.9 million by passive screening. In 2021 and 2022 the elimination of HAT as a public health problem was validated in Benin, Uganda, Equatorial Guinea and Ghana for gambiense HAT and in Rwanda for rhodesiense HAT. To reach the next goal of elimination of transmission of gambiense HAT, countries have to report zero cases of human infection with T. b. gambiense for a period of at least 5 consecutive years. The criteria and procedures to verify elimination of transmission have been recently published by WHO. Conclusions/significance: HAT elimination as a public health problem has been reached at global level, with seven countries already validated as having reached this goal. This achievement was made possible by the work of NSSCPs, supported by different public and private partners, and coordinated by WHO. The new challenging goal now is to reach zero cases by 2030. To reach this goal is crucial to maintain the engagement and support of donors and stakeholders and to keep the involvement and coordination of all partners. Along with the focus on elimination of transmission of gambiense HAT, it is important not to neglect rhodesiense HAT, which is targeted for elimination as a public health problem in the WHO road map for NTDs 2021–2030. Author summary: Human African trypanosomiasis (HAT), also known as "sleeping sickness, is a neglected tropical disease (NTD) transmitted by the bite of infected tsetse flies mainly affecting rural areas in sub-Saharan Africa. During the 20th century it caused enormous suffering in the endemic areas with a last reported peak in the late 1990s. This situation triggered a successful coordinated control effort that allowed, in 2012, the targeting of elimination of the disease as a public health problem by 2020. This paper follows previous papers to present the achievements towards HAT elimination. Annually reported cases have been kept below 1000 cases per year. HAT surveillance is maintained through active screening in endemic areas but also within a network of health facilities able to detect the disease. The World Health Organization (WHO), as defined in its 2021–2030 road map for NTDs, is now targeting the elimination of transmission of gambiense HAT and the elimination of rhodesiense HAT as a public health problem. The commitment of national health authorities and the international community will be essential to achieve these ambitious targets. [ABSTRACT FROM AUTHOR]

Background: Dengue virus (DENV) is endemic to many parts of the world and has serious health and socioeconomic effects even in high-income countries, especially with rapid changes in the climate globally. We explored the literature on dengue vector control methods used in high-income, city settings and associations with dengue incidence, dengue prevalence, or mosquito vector densities. Methods: Studies of any design or year were included if they reported effects on human DENV infection or Aedes vector indices of dengue-specific vector control interventions in high-income, city settings. Results: Of 24 eligible sources, most reported research in the United States (n = 8) or Australia (n = 5). Biocontrol (n = 12) and chemical control (n = 13) were the most frequently discussed vector control methods. Only 6 sources reported data on the effectiveness of a given method in reducing human DENV incidence or prevalence, 2 described effects of larval and adult control on Aedes DENV positivity, 20 reported effectiveness in reducing vector density, using insecticide, larvicide, source reduction, auto-dissemination of pyriproxyfen and Wolbachia, and only 1 described effects on human-vector contact. Conclusions: As most studies reported reductions in vector densities, rather than any effects on human DENV incidence or prevalence, we can draw no clear conclusions on which interventions might be most effective in reducing dengue in high-income, city areas. More research is needed linking evidence on the effects of different DENV vector control methods with dengue incidence/prevalence or mosquito vector densities in high-income, city settings as this is likely to differ from low-income settings. This is a significant evidence gap as climate changes increase the global reach of DENV. The importance of community involvement was clear in several studies, although it is impossible to tease out the relative contributions of this from other control methods used. Author summary: Dengue virus is present in many parts of the world and has serious health and socioeconomic effects even in high-income countries, especially with rapid changes in the climate globally. In this study, we explored the literature on dengue vector control methods used in high-income, city settings and associations with number of human dengue infections, and the density of mosquitoes. We found 24 papers with relevant results. Most of these described studies in the United States or Australia. Most were about various forms of biological or chemical control of the mosquitoes. Few papers discussed effects on human dengue infection. We were unable to draw any clear conclusions on how effective mosquito control methods were as we could identify little research on this subject in this specific setting. More research is needed on this topic, particularly as climate change will make more areas of the world vulnerable to dengue infection. [ABSTRACT FROM AUTHOR]

Madurella mycetomatis is the main cause of mycetoma, a chronic granulomatous infection for which currently no adequate therapy is available. To improve therapy, more knowledge on a molecular level is required to understand how M. mycetomatis is able to cause this disease. However, the genetic toolbox for M. mycetomatis is limited. To date, no method is available to genetically modify M. mycetomatis. In this paper, a protoplast-mediated transformation protocol was successfully developed for this fungal species, using hygromycin as a selection marker. Furthermore, using this method, a cytoplasmic-GFP-expressing M. mycetomatis strain was created. The reported methodology will be invaluable to explore the pathogenicity of M. mycetomatis and to develop reporter strains which can be useful in drug discovery as well as in genetic studies. Author summary: Madurella mycetomatis is the main cause of mycetoma, a Neglected Tropical Disease characterized by large tumorous swellings. Currently little is known about the role of the pathogen in mycetoma formation. Furthermore, there are only a few drugs which are able to inhibit this fungus. In order to gain insight in the disease mycetoma and identify novel drug targets we need a method to genetically modify M. mycetomatis. Up till now this was not available for M. mycetomatis. In this paper, we developed a method in which we for the first time were able to genetically modify Madurella mycetomatis. We did this by removing the M. mycetomatis cell wall and introduced a plasmid into the fungal cell which made the fungus express GFP. The reported methodology can be adapted to introduce other reporters in M. mycetomatis and will be invaluable to explore the pathogenicity of M. mycetomatis. Key Points: A genetic transformation method for Madurella mycetomatis has been developed. Hygromycin is a suitable selection marker for Madurella mycetomatis. GFP can be used as a reporter for Madurella mycetomatis. [ABSTRACT FROM AUTHOR]

Background: Pythiosis is a rare disease with high mortality, with over 94% of cases reported from Thailand and India. Prompt diagnosis and surgery improves patient outcomes. Therefore, continuing professional development (CPD) is essential for early recognition. However, a needs assessment related to a pythiosis CPD program has not been performed. Objectives: We conducted a needs assessment to develop a pythiosis CPD program. Patients/Methods: We conducted a survey study with 267 King Chulalongkorn Memorial Hospital residents (141 internal medicine (IM) residents and 126 surgery residents). A 30-item survey consisting of a knowledge assessment, demographic section, and an attitudes portion was distributed both electronically and via paper. The data was analyzed with descriptive and inferential statistics. Results: Sixty-seven percent completed the survey (110/141 IM residents, 70/126 surgery residents). The mean score [95% confidence interval] on the knowledge assessment was 41.67% [39.64%-43.69%] across all objectives. The three domains with the highest scores were pythiosis risk factors (67.22% correct), microbiologic characteristics (50.83%), and radiographic interpretation (50.56%). The three domains with the lowest scores were laboratory investigation (15.00%), epidemiology (29.17%), and symptomatology (30.83%). Most participants noted that the program should be online with both synchronous and asynchronous sessions, with a preferred length of 60–90 minutes per session. Conclusion: The pythiosis CPD program should emphasize education regarding symptomatology, laboratory investigation, and epidemiology, all of which are critical for the early detection of pythiosis to decrease mortality from this devastating disease. Most respondents felt this program was necessary and should be implemented in a virtual blended format. Author summary: Researchers conducted a needs assessment to develop a Continuing Professional Development (CPD) program on pythiosis, a rare and often fatal disease prevalent in Thailand and India. The study surveyed 267 residents at King Chulalongkorn Memorial Hospital, revealing a 67% completion rate. The residents exhibited a mean knowledge score of 41.67%, with the highest proficiency in pythiosis risk factors (67.22%), microbiologic characteristics (50.83%), and radiographic interpretation (50.56%). Conversely, lower scores were observed in laboratory investigation (15.00%), epidemiology (29.17%), and symptomatology (30.83%). Participants expressed a preference for an online CPD program with both synchronous and asynchronous sessions, each lasting 60–90 minutes. The study emphasizes the need for education on symptomatology, laboratory investigation, and epidemiology to enhance early pythiosis detection and reduce mortality. Respondents widely supported the implementation of the proposed virtual blended CPD program. [ABSTRACT FROM AUTHOR]

Rift Valley fever (RVF) is a neglected vector-borne disease which is endemic in many countries across Africa and has seen recent geographical expansions into the Arabian Peninsula. RVF can cause severe infections in both animals and humans. RVF infections in livestock can lead to mass fatalities. In humans, the symptoms are nonspecific and can often lead to misdiagnosis. However, a small proportion progresses to haemorrhagic infection with a significantly higher mortality rate. The culmination of this can cause severe socioeconomic impacts. This review aims to identify the main socioeconomic impacts caused by RVF outbreaks as well as existing knowledge gaps. Ninety-three academic and grey papers were selected, covering 19 countries and 10 methodological approaches. A variety of socioeconomic impacts were found across all levels of society: Livestock trade disruptions consequently impacted local food security, local and national economies. Most livestock farmers in endemic countries are subsistence farmers and so rely on their livestock for sustenance and income. RVF outbreaks resulted in a variety of socioeconomic impacts, e.g., the inability to pay for school fees. Main barriers to vaccine uptake in communities were lack of access, funds, interest along with other social aspects. The occupational risks for women (and pregnant women) are largely unknown. To our knowledge, this is the first review on RVF to highlight the clear knowledge gap surrounding the potential gender differences on risks of RVF exposure, as well as differences on occupational health risk in pastoral communities. Further work is required to fill the gaps identified in this review and inform control policies. Author summary: Rift Valley fever (RVF) outbreaks have affected many regions globally with varying severity. The number of reported cases in animals and humans is patchy at best, with some countries not recording animal events. This review focuses on 3 main themes: Health Burden; Economic Impacts; and Prevention and Surveillance. The review explores socioeconomic impacts across local, regional, and national levels. We found women have varying degrees of risk to exposure of RVF; however, more research is required to better understand gender differences. RVF outbreaks tend to begin in livestock and spillover into humans. The complex disease dynamics are poorly understood because of the interactions between animals, humans, and the environment. This cross-boundary dynamics fall outside of the public health remit and into veterinary health. This allows for outbreaks to spread uncontrollably for some time. More research using a One Health Lens is required which explores these complex dynamics with the combination of different social aspects. This review highlights the wider impacts experienced at different levels of society and where there are major gaps in our knowledge. Further analysis into these gaps is required to and will aid policy makers in developing new control measures around RVF. [ABSTRACT FROM AUTHOR]

Background: Localized cutaneous leishmaniasis (LCL) is a chronic ulcerating disease. A literature review identified inconsistencies in clinical trials. The aims of this study were to reach a consensus on the most important domains to measure when assessing LCL, agree on parameters to measure the domains, and develop a tool representing a Core Outcome Set (COS), for use in clinical assessment of LCL. Methodology & principal findings: A literature review was conducted to identify any existing COS for LCL embracing agreed Outcome Domains, i.e. what to measure and any Outcome Measurement Instruments (OMIs). As no COS was available, potential outcome domains for assessment of LCL were identified through an international collaborative approach using e-consultations and virtual discussions with expert stakeholders (n = 20) from geographically different LCL endemic countries. Subsequent judgmental validation process included a face-to-face multidisciplinary stakeholders' meeting adopting the Nominal Group Technique. A final consensual agreement on outcome domains and items required to measure these domains was established. "Clinical Cure" was defined as the ideal overall "General Concept". The five Core Outcome Domains included Signs capturing clinical morphology, diameter, and induration of an index lesion with the aid of a palpability score, Treatment Efficacy assessing percentage change in size of the lesion and re-epithelialization compared to baseline, Treatment Impact which included an investigator and patient visual analogue score, and Clinical Sequelae rating pigment change, atrophic and hypertrophic/keloid scars. It was agreed that two open-ended questions should be included to capture some aspects of Health-Related Quality of Life as a means of capturing a patient-focused approach. Conclusion: LeishCOM_LCL was generated to reflect a COS for LCL. This captured demographic details, agreed outcome domains and measures to assess these domains. Validation of LeishCOM_LCL will be reported in a separate paper. Development of a Patient Reported Outcome Measure will be considered in the future. Author summary: Localized cutaneous leishmaniasis (LCL) is a chronic ulcerating disease caused by the parasite Leishmania spp. Literature review identified inconsistencies in methods and parameters used to evaluate treatment/alternative-interventions resulting in difficulties in comparing new treatment/interventions in clinical trials. In our international consensual study, we adopted the face-to-face nominal group technique and a judgment process to identify domains key to assessment of LCL. Subsequent measures for each domain were used to form a Core Outcome Set (COS). LeishCOM_LCL was developed as an outcome measure instrument (OMI) to capture the COS incorporating existing and newly developed tools. "Clinical Cure" was agreed as the "General Concept" to be captured through five domains. It was agreed that "Signs" domain should capture clinical morphology, diameter, and induration of an index lesion with the aid of a palpability score. "Treatment Efficacy" was assessed by recording percentage change in size of the lesion and re-epithelialization compared to baseline. "Treatment Impact" was reflected through an investigator and the patient visual analogue score and "Clinical Sequelae" rated pigment change, atrophic/hypertrophic scars. Two open-ended questions were included to capture some aspects of "Health-Related Quality of Life". LeishCOM_LCL also records patient demographic details and was validated in a small cohort of patients. [ABSTRACT FROM AUTHOR]

American tegumentary leishmaniasis (ATL) is highly endemic in the Amazon basin and occurs in all South American countries, except Chile and Uruguay. Most Brazilian ATL cases are due to Leishmania (Viannia) braziliensis, however other neglected Amazonian species are being increasingly reported. They belong to the subgenus L. (Viannia) and information on suitable models to understand immunopathology are scarce. Here, we explored the use of the golden hamster Mesocricetus auratus and its macrophages as a model for L. (Viannia) species. We also studied the interaction of parasite glycoconjugates (LPGs and GIPLs) in murine macrophages. The following strains were used: L. (V.) braziliensis (MHOM/BR/2001/BA788), L. (V.) guyanensis (MHOM/BR/85/M9945), L. (V.) shawi (MHOM/BR/96/M15789), L. (V.) lindenbergi (MHOM/BR/98/M15733) and L. (V.) naiffi (MDAS/BR/79/M5533). In vivo infections were initiated by injecting parasites into the footpad and were followed up at 20- and 40-days PI. Parasites were mixed with salivary gland extract (SGE) from wild-captured Nyssomyia neivai prior to in vivo infections. Animals were euthanized for histopathological evaluation of the footpads, spleen, and liver. The parasite burden was evaluated in the skin and draining lymph nodes. In vitro infections used resident peritoneal macrophages and THP-1 monocytes infected with all species using a MOI (1:10). For biochemical studies, glycoconjugates (LPGs and GIPLs) were extracted, purified, and biochemically characterized using fluorophore-assisted carbohydrate electrophoresis (FACE). They were functionally evaluated after incubation with macrophages from C57BL/6 mice and knockouts (TLR2-/- and TLR4-/-) for nitric oxide (NO) and cytokine/chemokine production. All species, except L. (V.) guyanensis, failed to generate evident macroscopic lesions 40 days PI. The L. (V.) guyanensis lesions were swollen but did not ulcerate and microscopically were characterized by an intense inflammatory exudate. Despite the fact the other species did not produce visible skin lesions there was no or mild pro-inflammatory infiltration at the inoculation site and parasites survived in the hamster skin/lymph nodes and even visceralized. Although none of the species caused severe disease in the hamster, they differentially infected peritoneal macrophages in vitro. LPGs and GIPLs were able to differentially trigger NO and cytokine production via TLR2/TLR4 and TLR4, respectively. The presence of a sidechain in L. (V.) lainsoni LPG (type II) may be responsible for its higher proinflammatory activity. After Principal Component analyses using all phenotypic features, the clustering of L. (V.) lainsoni was separated from all the other L. (Viannia) species. We conclude that M. auratus was a suitable in vivo model for at least four dermotropic L. (Viannia) species. However, in vitro studies using peritoneal cells are a suitable alternative for understanding interactions of the six L. (Viannia) species used here. LRV1 presence was found in L. (V.) guyanensis and L. (V.) shawi with no apparent correlation with virulence in vitro and in vivo. Finally, parasite glycoconjugates were able to functionally trigger various innate immune responses in murine macrophages via TLRs consistent with their inflammatory profile in vivo. Author summary: American Tegumentary Leishmaniasis (ATL) is one of the major neglected diseases occurring in Brazil and the Amazon contributes with a wide range of species that are not completely known. These little-known species are transmitted to rural workers entering the forest during lumbering, mining, and latex extraction activities and rarely in peri domestic situation. Infection occurs by the bite of infected sand flies in different environments and biomes. One of the major constraints in studying L. (Viannia) species is the difficulty of finding suitable animal and cellular models to investigate their biology. The mouse model is not a suitable model for most L. (Viannia) species. However, hamsters were considered a good model for L. (V.) braziliensis. In the search for a better model for some neglected dermotropic Amazonian Leishmania, we evaluated interactions in vivo in the hamster and in vitro in its macrophages. All species infected cells in vitro, but hamsters could only be considered as a suitable in vivo model for L. (V.) guyanensis. Importantly the glycoconjugates of the six L. (Viannia) species were very proinflammatory, showing various levels of macrophage activation. This paper opens possibilities of determining more accurately the taxonomic position of L. (V.) lainsoni within the subgenus L. (Viannia). [ABSTRACT FROM AUTHOR]

Background: Paragonimiasis, primarily caused by Paragonimus westermani and P. skrjabini in China, is a common food-borne parasitic zoonosis. However, the national distribution of Paragonimus spp. infection and its associated environmental determinants remain poorly understood. In this paper, we summarize the infection of P. westermani and P. skrjabini and describe key biogeographical characteristics of the endemic areas in China. Methods: Data on Paragonimus infection in humans and animal hosts were extracted from eight electronic databases, including CNKI, CWFD, Chongqing VIP, SinoMed, Medline, Embase, PubMed, and Web of Science. A random-effects meta-analysis model was used to estimate the pooled prevalence. All survey locations were georeferenced and plotted on China map, and scatter plots were used to illustrate the biogeographical characteristics of regions reporting Paragonimus infection. Results: A total of 28,948 cases of human paragonimiasis have been documented, with 2,401 cases reported after 2010. Among the 11,443 cases with reported ages, 88.05% were children or adolescents. The pooled prevalence of P. skrjabini is 0.45% (95% CI: 0.27–0.66%) in snails, 31.10% (95% CI: 24.77–37.80%) in the second intermediate host, and 20.31% (95% CI: 9.69–33.38%) in animal reservoirs. For P. westermani, the pooled prevalence is 0.06% (95% CI: 0.01–0.13%) in snails, 52.07% (95% CI: 43.56–60.52%) in the second intermediate host, and 21.40% (95% CI: 7.82–38.99%) in animal reservoirs. Paragonimus are primarily distributed in regions with low altitude, high temperature, and high precipitation. In northeastern China, only P. westermani infections have been documented, while in more southern areas, infections of both P. westermani and P. skrjabini have been reported. Conclusions: Paragonimiasis remains prevalent in China, particularly among children and adolescents. Variations exist in the intermediate hosts and geographical distribution of P. westermani and P. skrjabini. Additionally, altitude, temperature, and precipitation may influence the distribution of Paragonimus. Author summary: Paragonimiasis, a foodborne zoonotic parasitic disease caused by lung flukes (Paragonimus spp.), remains a significant neglected public health threat in many Asian countries, including China. Human infection occurs through the ingestion of raw or undercooked freshwater crab or crayfish containing the metacercariae stage. Given the popularity of consuming raw or undercooked freshwater products in many areas of China, understanding the infection status and spatial distribution of Paragonimus spp. in humans and animal hosts is crucial for controlling paragonimiasis. Our study provides a comprehensive summary of the infection levels of the two most important zoonotic Paragonimus species, P. westermani and P. skrjabini, in humans and animal hosts in China, along with a description of the spatial distribution and environmental characteristics of their endemic areas. We observe a wide distribution of Paragonimus infection in China, with a significant prevalence found in freshwater crabs and crayfish. Our findings underscore the importance of avoiding the consumption of raw or undercooked freshwater products to prevent foodborne diseases, including paragonimiasis. [ABSTRACT FROM AUTHOR]

Overview: The roadmap adopted by the World Health Organization (WHO) for eliminating neglected tropical diseases aims to eliminate schistosomiasis, as a public health concern, by 2030. While progress has been made towards reducing schistosomiasis morbidity control in several sub-Saharan African countries, there is still more that needs to be done. Proper surveillance using accurate diagnostics with acceptable sensitivity and specificity is essential for evaluating the success of all efforts against schistosomiasis. Microscopy, despite its low sensitivity, remains the gold standard approach for diagnosing the disease. Although many efforts have been made to develop new diagnostics based on circulating parasite proteins, genetic markers, schistosome egg morphology, and their paramagnetic properties, none has been robust enough to replace microscopy. This review highlights common diagnostic approaches for detecting schistosomiasis in field and clinical settings, major challenges, and provides new and novel opportunities and diagnosis pathways that will be critical in supporting elimination of schistosomiasis. Methods: We searched for relevant and reliable published literature from PubMed, Scopus, google scholar, and Web of science. The search strategies were primarily determined by subtopic, and hence the following words were used (schistosom*, diagnosis, Kato–Katz, antibody test, circulating antigen, POC-CCA, UCP-LF-CAA, molecular diagnostics, nucleic acid amplification test, microfluidics, lab-on a disk, lab-on chip, recombinase polymerase amplification (RPA), LAMP, portable sequencer, nanobody test, identical multi-repeat sequences, diagnostic TPPs, REASSURED, extraction free), and Boolean operators AND and/OR were used to refine the searching capacity. Due to the global public health nature of schistosomiasis, we also searched for reliable documents, reports, and research papers published by international health organizations, World Health Organization (WHO), and Center for Disease control and Elimination. [ABSTRACT FROM AUTHOR]

Background: Ticks, as critical vectors of a variety of pathogens, pose a significant public health challenge globally. In Southeast Asia (SEA), ticks are responsible for transmitting a diverse array of pathogens affecting humans and animals. The geographical and ecological diversity of SEA provides a unique environment that supports a wide range of tick species, which complicates the management and study of tick-borne diseases (TBDs). Methodology/principal findings: This article synthesizes findings from the first international symposium on ticks and TBDs in Southeast Asia, held in Phnom Penh on June 22 and 23, 2023. It highlights regional efforts to understand tick ecology and pathogen transmission. This paper proposes to present a summary of the various presentations given during the symposium following 3 main parts. The first one is devoted to the state of knowledge regarding ticks and TBDs in SEA countries, with presentations from 6 different countries, namely Cambodia, Indonesia, Laos, Malaysia, Thailand, and Vietnam. The second part focuses on the development of new research approaches on tick-borne pathogens (TBPs) and TBDs. The last part is a summary of the round table discussion held on the final day, with the aim of defining the most important challenges and recommendations for researches on TBP and TBD in the SEA region. Conclusions/Significance: Key topics discussed include advancements in diagnostic tools, such as MALDI-TOF MS and proteomics, and the development of sustainable strategies for tick management and disease prevention. The symposium facilitated the exchange of knowledge and collaborative networks among experts from various disciplines, promoting a unified approach to tackling TBDs in the region. The symposium underscored the need for enhanced surveillance, diagnostics, and inter-regional cooperation to manage the threat of TBDs effectively. Recommendations include the establishment of a regional database for tick identification and the expansion of vector competence studies. These initiatives are crucial for developing targeted interventions and understanding the broader implications of climate change and urbanization on the prevalence of TBDs. Author summary: Ticks are arthropods that can transmit pathogens affecting both animals and people. In Southeast Asia, the diversity of tick species and the tick-borne pathogens is particularly high due to the region's varied climates and ecosystems. Our research, discussed during an international symposium, focuses on understanding these ticks and finding better ways to detect and manage the associated diseases. This symposium highlighted knowledge on ticks and tick-borne pathogens in the different countries, new technologies such as perspectives in research. This work is important because it helps in developing strategies to prevent these diseases, which is crucial for the health of people and animals in Southeast Asia and beyond. This work not only advances scientific knowledge but also supports efforts to improve public health in response to the challenges posed by environmental changes, such as climate and land-use changes. [ABSTRACT FROM AUTHOR]

Background: Dengue, Zika, and chikungunya, whose viruses are transmitted mainly by Aedes aegypti, significantly impact human health worldwide. Despite the recent development of promising vaccines against the dengue virus, controlling these arbovirus diseases still depends on mosquito surveillance and control. Nonetheless, several studies have shown that these measures are not sufficiently effective or ineffective. Identifying higher-risk areas in a municipality and directing control efforts towards them could improve it. One tool for this is the premise condition index (PCI); however, its measure requires visiting all buildings. We propose a novel approach capable of predicting the PCI based on facade street-level images, which we call PCINet. Methodology: Our study was conducted in Campinas, a one million-inhabitant city in São Paulo, Brazil. We surveyed 200 blocks, visited their buildings, and measured the three traditional PCI components (building and backyard conditions and shading), the facade conditions (taking pictures of them), and other characteristics. We trained a deep neural network with the pictures taken, creating a computational model that can predict buildings' conditions based on the view of their facades. We evaluated PCINet in a scenario emulating a real large-scale situation, where the model could be deployed to automatically monitor four regions of Campinas to identify risk areas. Principal findings: PCINet produced reasonable results in differentiating the facade condition into three levels, and it is a scalable strategy to triage large areas. The entire process can be automated through data collection from facade data sources and inferences through PCINet. The facade conditions correlated highly with the building and backyard conditions and reasonably well with shading and backyard conditions. The use of street-level images and PCINet could help to optimize Ae. aegypti surveillance and control, reducing the number of in-person visits necessary to identify buildings, blocks, and neighborhoods at higher risk from mosquito and arbovirus diseases. Author summary: The strategies to control Ae. aegypti require intensive work and considerable financial resources, are time-consuming, and are commonly affected by operational problems requiring urgent improvement. The PCI is a good tool for identifying higher-risk areas; however, its measure requires a high amount of human and material resources, and the aforementioned issues remain. In this paper, we propose a novel approach capable of predicting the PCI of buildings based on street-level images. This first work combines deep learning-based methods with street-level data to predict facade conditions. Considering the good results obtained with PCINet and the good correlations of facade conditions with PCI components, we could use this methodology to classify building conditions without visiting them physically. With this, we intend to overcome the high cost of identifying high-risk areas. Although we have a long road ahead, our results show that PCINet could help to optimize Ae. aegypti and arbovirus surveillance and control, reducing the number of in-person visits necessary to identify buildings or areas at risk. [ABSTRACT FROM AUTHOR]

Chagas disease is caused by Trypanosoma cruzi, a protozoan parasite that displays considerable genetic diversity. Infections result in a range of pathological outcomes, and different strains can exhibit a wide spectrum of anti-parasitic drug tolerance. The genetic determinants of infectivity, virulence and therapeutic susceptibility remain largely unknown. As experimental tools to address these issues, we have generated a panel of bioluminescent:fluorescent parasite strains that cover the diversity of the T. cruzi species. These reporters allow spatio-temporal infection dynamics in murine models to be monitored in a non-invasive manner by in vivo imaging, provide a capability to detect rare infection foci at single-cell resolution, and represent a valuable resource for investigating virulence and host:parasite interactions at a mechanistic level. Importantly, these parasite reporter strains can also contribute to the Chagas disease drug screening cascade by ensuring that candidate compounds have pan-species in vivo activity prior to being advanced into clinical testing. The parasite strains described in this paper are available on request. Author summary: Chagas disease results from infection with the protozoan parasite Trypanosoma cruzi and is a major public health problem throughout Latin America. T. cruzi is a genetically diverse species and infection can result in a wide range of pathological outcomes, mainly associated with the heart and/or digestive tract. Research on Chagas disease, ranging from fundamental biology to drug development, has been greatly aided by the availability of genetically modified parasite reporter strains that express bioluminescent:fluorescent fusion proteins. In combination with mouse models and imaging technology, these strains allow infections to be monitored in real-time, with high sensitivity, and infection foci to be visualised at single-cell resolution. Here, we describe an extensive panel of bioluminescent and fluorescent strains that cover the diversity of the T. cruzi species. These reporter strains, that are available on request, should have wide utility in many areas of Chagas disease research. In particular, as part of the drug development screening programme, they can be used to ensure that candidate compounds have in vivo activity across the species prior to being advanced into clinical testing. [ABSTRACT FROM AUTHOR]

Critical scientific questions remain regarding infection with Mycobacterium ulcerans, the organism responsible for the neglected tropical disease, Buruli ulcer (BU). A controlled human infection model has the potential to accelerate our knowledge of the immunological correlates of disease, to test prophylactic interventions and novel therapeutics. Here we present microbiological evidence supporting M. ulcerans JKD8049 as a suitable human challenge strain. This non-genetically modified Australian isolate is susceptible to clinically relevant antibiotics, can be cultured in animal-free and surfactant-free media, can be enumerated for precise dosing, and has stable viability following cryopreservation. Infectious challenge of humans with JKD8049 is anticipated to imitate natural infection, as M. ulcerans JKD8049 is genetically stable following in vitro passage and produces the key virulence factor, mycolactone. Also reported are considerations for the manufacture, storage, and administration of M. ulcerans JKD8049 for controlled human infection. Non-technical author summary: In this paper, we progress our research towards establishing a controlled human infection model (CHIM) of Buruli ulcer, caused by Mycobacterium ulcerans. It is crucial that the challenge agent used for such experiments is thoroughly characterised; not only is this important to ensure standardisation between participants, but also to reinforce their safety. Using established criteria, we systematically demonstrate why the isolate M. ulcerans JKD8049 may be a suitable challenge strain. This strain is non-genetically modified, susceptible to antibiotics, can be cultured in relatively simple media, and can be filtered and preserved as individual bacteria to ensure accurate dosing of participants. Even after repeatedly culturing the organism for months, it remains genetically stable, so significant mutations are not expected to develop during manufacture. After preserving the isolate at -80°C, it can be revived and diluted for human inoculation at very low doses. Importantly, it produces the toxin mycolactone, so it is expected to recreate typical Buruli ulcer disease in participants, which will be important to properly test vaccines and other therapeutics. We also demonstrate a set of principles to ensure the produced batch is free from contamination. [ABSTRACT FROM AUTHOR]

Background: Mozambique has one of the highest burdens of neglected tropical diseases in Africa. Lymphatic filariasis, schistosomiasis and soil-transmitted helminths are being targeted for elimination as part of integrated mass drug administration campaigns. The progress made towards interruption of transmission has been affected by recent conflict in Cabo Delgado province. The aim of this paper was to determine the potential impact of this crisis on the neglected tropical diseases programme and the challenges in reaching the elimination goals of 2030. Methodology: A desk-based secondary data analysis was conducted on publicly available sources of neglected tropical diseases, conflict incidents, internally displaced persons and geographical access between 2020 and 2022. Data were summarised and mapped using GIS software. A combined risk stratified assessment at district level was developed with five classifications i) Very high-risk; ii) High-risk; iii) Medium to high-risk; iv) Medium risk; and v) Not at risk due to conflict absence but co-endemic. Results: Lymphatic filariasis, schistosomiasis and soil-transmitted helminths were co-endemic in 115 out of 156 (74%) districts. Between 2020 and 2022 a total of 1,653 conflict-related incidents were reported, most of them in Cabo Delgado province (n = 1,397, 85%). A five-fold increase of internally displaced persons was recorded from April 2020 (n = 172,186) to November 2022 (n = 935,130). Geographical accessibility also deteriorated across the province with an increase from five (29%) in 2021 to seven (41%) districts in 2022 classified as hard-to-reach. The combined risk stratification identified that most districts (n = 7; 41%) in Cabo Delgado province had medium to high-risk; very high-risk (n = 5, 29%); medium risk (n = 3, 18%); high-risk (n = 2, 12%). Conclusion: Most of the districts of Cabo Delgado were considered to be at risk of not meeting the neglected tropical diseases road map 2021–2030 targets due to the humanitarian crisis ongoing. There is the need for practical strategies and funding to overcome these hostile challenges. Author summary: Neglected tropical diseases (NTDs) are a group of diseases that usually affect the most vulnerable population in tropical countries. Mozambique has been tackling lymphatic filariasis, schistosomiasis and soil-transmitted helminths for two decades with the aim of eliminating them through large-scale distribution of drugs to interrupt transmission. In recent years, internal conflict has emerged and led to an escalating humanitarian crisis in the endemic province of Cabo Delgado which has hampered elimination efforts. This study aimed to shed light on the situation and to determine the potential impact on the Mozambique NTDs' programme. We used a range of open-source data on NTDs, conflict, internally displaced persons and geographical access to summarise, describe, and map the situation at the district level in the most affected province. We used this data to also examine their geographical overlap and to develop a risk assessment tool to highlight the high risk and most vulnerable areas. We found that between 2021–2022 more than 1,300 conflict incidents occurred, and almost 1 million people were displaced in Cabo Delgado province with many districts having limited geographical access which impacted the NTDs programme's ability to work. The study highlights the important challenges of conflict and displacement of vulnerable populations, and the need for support and set of guidelines for NTDs endemic programmes to use to help to overcome key barriers and meet the targets of elimination set by the World Health Organization for 2030. [ABSTRACT FROM AUTHOR]

Background: Lymphatic filariasis (LF) is a parasitic disease transmitted by mosquitoes, causing severe pain, disfiguring, and disabling clinical conditions such as lymphoedema and hydrocoele. LF is a global public health problem affecting 72 countries, primarily in Africa and Asia. Since 2000, the World Health Organization (WHO) has led the Global Programme to Eliminate Lymphatic Filariasis (GPELF) to support all endemic regions. This paper focuses on the achievements of the Malawi LF Elimination Programme between 2000 and 2020 to eliminate LF as a public health problem, making it the second sub-Saharan country to receive validation from the WHO. Methodology/principal findings: The Malawi LF Programme addressed the widespread prevalence of LF infection and disease across the country, using the recommended WHO GPELF strategies and operational research initiatives in collaboration with key national and international partners. First, to stop the spread of infection (i.e., interrupt transmission) and reduce the circulating filarial antigen prevalence from as high as 74.4% to below the critical threshold of 1–2% prevalence, mass drug administration (MDA) using a two-drug regime was implemented at high coverage rates (>65%) of the total population, with supplementary interventions from other programmes (e.g., malaria vector control). The decline in prevalence was monitored and confirmed over time using several impact assessment and post-treatment surveillance tools including the standard sentinel site, spot check, and transmission assessment surveys and alternative integrated, hotspot, and easy-access group surveys. Second, to alleviate suffering of the affected populations (i.e., control morbidity) the morbidity management and disability prevention (MMDP) package of care was implemented. Specifically, clinical case estimates were obtained via house-to-house patient searching activities; health personnel and patients were trained in self-care protocols for lymphoedema and/or referrals to hospitals for hydrocoele surgery; and the readiness and quality of treatment and services were assessed with new survey tools. Conclusions: Malawi's elimination of LF will ensure that future generations are not infected and suffer from the disfiguring and disabling disease. However, it will be critical that the Malawi LF Elimination programme remains vigilant, focussing on post-elimination surveillance and MMDP implementation and integration into routine health systems to support long-term sustainability and ongoing success. Summary: Lymphatic filariasis, also known as elephantiasis, is a disabling, disfiguring, and painful disease caused by a parasite that infected mosquitoes transmit to millions of people worldwide. Since 2000, the Global Programme to Eliminate Lymphatic Filariasis (GPELF) has supported endemic countries such as Malawi in south-eastern Africa, to eliminate the disease as a public health problem. The Malawi National LF Elimination Programme has worked tirelessly over the past two decades to implement the GPELF recommended strategies to interrupt the transmission with a two-drug regime, and to alleviate suffering in patients with lymphoedema and/or hydrocoele through morbidity management and disability prevention. Additionally, the LF Programme has collaborated with national and international stakeholders to implement a range of supplementary operational research projects to address outstanding knowledge gaps and programmatic barriers. In 2020, the World Health Organisation validated that Malawi had successfully eliminated LF as a public health problem, making it the second country in sub-Saharan Africa to achieve this, which is remarkable given that Malawi previously had very high infection rates. The LF Programme now remains vigilant, putting its efforts towards post-elimination surveillance and the continued implementation of care for patients with chronic conditions. Malawi's elimination of LF will ensure that future generations are not affected by this devastating disease. [ABSTRACT FROM AUTHOR]

Objectives: Deworming programmes of soil-transmitted helminths are generally monitored and evaluated by aggregating drug coverage and infection levels at a district level. However, heterogeneity in drug coverage at finer spatial scales means indicators may remain above thresholds for elimination as a public health problem or of transmission in some areas. This paper aims to highlight the misleading information that aggregating data at larger spatial scales can have for programme decision making. Methods: Drug coverage data from the Geshiyaro project were compared at two spatial scales with reference to the World Health Organisation's targets. District (woreda) and village (kebele) level were compared. The association between infection levels and drug coverage was analysed by fitting a weighted least-squares function to the mean intensity of infection (eggs per gram of faeces) against drug coverage. Results: The data show clearly that when the evaluation of coverage is aggregated to the district level, information on heterogeneity at a finer spatial scale is lost. Infection intensity decreases significantly (p = 0.0023) with increasing drug coverage. Conclusion: Aggregating data at large spatial scales can result in prematurely ceasing deworming, prompting rapid infection bounce-back. There is a strong need to define context-specific spatial scales for monitoring and evaluating intervention programmes. Author summary: For soil-transmitted helminths (STH), control is centred around mass drug administration (MDA) of eligible subgroups without prior diagnosis, aiming to achieve 75% drug coverage. MDA programmes are monitored by assessing whether drug coverage and infection prevalence and intensity, aggregated to a spatial scale, typically an administrative unit such as a district, have met programmatic targets. Spatial heterogeneity in drug coverage, prevalence, and intensity is commonplace and well documented. However, little attention has been paid to the spatial scales of this heterogeneity and the implications it has on providing the basis to inform the cessation or continuation of prophylactic mass treatment. We present a clear analysis showing that fine scale spatial heterogeneity in drug coverage is commonplace even in well implemented MDA programmes. We show that aggregating data at the district level, which is routine for deworming programmes, can give misleading information on which to base future treatment decisions, specifically the premature reduction in frequency, or cessation of MDA, because fine scale heterogeneity is not captured by larger aggregations. This work highlights the critical need to define context specific spatial scales for monitoring and evaluating interventions implemented in different transmission settings. [ABSTRACT FROM AUTHOR]

Introduction: Schistosomiasis is a parasitic disease in Tanzania affecting over 50% of the population. Current control strategies involve mass drug administration (MDA) campaigns at the district level, which have led to problems of over- and under-treatment in different areas. WHO guidelines have called for more targeted MDA to circumvent these problems, however a scarcity of prevalence data inhibits decision makers from prioritizing sub-district areas for MDA. This study demonstrated how geostatistics can be used to inform planning for targeted MDA. Methods: Geostatistical sub-district (ward-level) prevalence estimates were generated through combining a zero-inflated poisson model and kriging approach (regression kriging). To make predictions, the model used prevalence survey data collected in 2021 of 17,400 school children in six regions of Tanzania, along with several open source ecological and socio-demographic variables with known associations with schistosomiasis. Results: The model results show that regression kriging can be used to effectively predict the ward level parasite prevalence of the two species of Schistosoma endemic to the study area. Kriging was found to further improve the regression model fit, with an adjusted R-squared value of 0.51 and 0.32 for intestinal and urogenital schistosomiasis, respectively. Targeted treatment based on model predictions would represent a shift in treatment away from 193 wards estimated to be over-treated to 149 wards that would have been omitted from the district level MDA. Conclusions: Geostatistical models can help to support NTD program efficiency and reduce disease transmission by facilitating WHO recommended targeted MDA treatment through provision of prevalence estimates where data is scarce. Author summary: In Tanzania, schistosomiasis is a vast public health problem treated through mass drug administration (MDA) campaigns that target large groups of the population. Such mass drug administration (MDA) campaigns require significant amounts of resources challenging the capacity of chronically underfunded schistosomiasis control programs to sustain annually. The way in which MDA campaigns have been conducted, by targeting whole districts believed to be endemic, have not had optimal results for reducing disease transmission, have facilitated problems of under-treating in many population groups while over-treating other non-endemic areas, and have placed a significant strain on the limited resources available. To circumvent such problems, the World Health Organization (WHO) has recommended that treatment efforts need to be more targeted to endemic communities and administrative areas within districts, and that MDA campaigns should be conducted at a sub-district level. The WHO also provides explicit guidelines for treatments of communities based on parasitological prevalence of schistosomiasis. While there is sound rationale in making this switch, most schistosomiasis endemic countries including Tanzania do not have adequate surveillance data to make such informed decisions at this level. Limited data on schistosomiasis prevalence inhibits disease control programs from making data informed decision on where to treat, as well as adhering to WHO recommendations for conducting targeted treatment at the sub-district level. Geostatistical models are spatial analysis tools that have been used in the past to help predict the likelihood of disease prevalence in nearby areas where there is limited data. They are particularly helpful in predicting diseases like schistosomiasis that have strong associations with environmental and socio-demographic elements that we do have data on widespread across much of the world. Publications on the use geostatistical models to predict schistosomiasis prevalence in different settings date back more than 20 years, but have not yet been integrated to aid the decision process of making of national programs, nor have they been specifically advocated for by the WHO. The study seeks to examine the implications of using model predictions to guide targeted, sub-district level treatment, in terms of populations requiring treatments when compared with conventional district level approaches. This publication demonstrates that use of geostatistical models for predicting schistosomiasis is possible, can be a valuable tool to inform where to treat. It furthermore estimates that the populations eligible for targeted treatment are considerably different than the populations who would be receiving treatment under conventional district level treatment approached, placing emphasis on the importance of switching to targeted treatment. The paper argues that uptake of geostatistical models can enable targeted planning below the district level, a major step to facilitate a systematic change that the WHO believes will drive down transmission of the disease, the importance of which cannot be understated in an area that is so historically burdened by the disease and its associated health complications. [ABSTRACT FROM AUTHOR]

The geographical range of schistosomiasis is affected by the ecology of schistosome parasites and their obligate host snails, including their response to temperature. Previous models predicted schistosomiasis' thermal optimum at 21.7°C, which is not compatible with the temperature in sub-Saharan Africa (SSA) regions where schistosomiasis is hyperendemic. We performed an extensive literature search for empirical data on the effect of temperature on physiological and epidemiological parameters regulating the free-living stages of S. mansoni and S. haematobium and their obligate host snails, i.e., Biomphalaria spp. and Bulinus spp., respectively. We derived nonlinear thermal responses fitted on these data to parameterize a mechanistic, process-based model of schistosomiasis. We then re-cast the basic reproduction number and the prevalence of schistosome infection as functions of temperature. We found that the thermal optima for transmission of S. mansoni and S. haematobium range between 23.1–27.3°C and 23.6–27.9°C (95% CI) respectively. We also found that the thermal optimum shifts toward higher temperatures as the human water contact rate increases with temperature. Our findings align with an extensive dataset of schistosomiasis prevalence in SSA. The refined nonlinear thermal-response model developed here suggests a more suitable current climate and a greater risk of increased transmission with future warming for more than half of the schistosomiasis suitable regions with mean annual temperature below the thermal optimum. Author summary: In this research, we explored the complex interplay between temperature and the transmission risk of schistosomiasis, a parasitic disease currently affecting over two hundred million people, predominantly in SSA. We developed a novel mathematical model accounting for the multiple positive and negative ways temperature affects the free-living stages of the parasite and its obligate, non-human host, i.e., specific species of freshwater snails. Our models show that schistosomiasis transmission risk peaks at temperatures 1–6°C higher than previously estimated. This indicates that the impact of climate change on schistosomiasis transmission might be more extensive than previously thought, affecting a wide geographic range where mean annual temperatures are currently below the optimal temperature. Our model projections are consistent with the observed temperatures in locations of SSA where schistosomiasis is endemic and data on infection prevalence in the human population are available. These findings suggest that the current climate is conducive to schistosomiasis transmission, and future warming could escalate the risk further, emphasizing the need for targeted interventions in these regions. [ABSTRACT FROM AUTHOR]

The reported cases of scrub typhus (ST) have continued to escalate, with outbreaks occurring regionally in China. These pose an increasing public health threat at a time when public health has been overwhelmed. During the period from July to August 2022, in Rongjiang County, Guizhou Province, China, 13 out of 21 fever patients were diagnosed with scrub typhus, based on epidemiological investigation and blood test analysis. The major clinical symptoms of these patients showed fever, chills, headache, eschar, fatigue and pneumonia, which were accompanied by a rise in C-reactive protein, neutrophils, alanine transaminase (ALT) and aspartate aminotransferase (AST). Furthermore, nearly half of them exhibited abnormal electrocardiogram activity. Through semi-nested PCR, Sanger sequencing and phylogenetic tree construction, the Karp strain of Orientia tsutsugamushi (O. tsutsugamushi) was confirmed as the pathogen causing ST in Rongjiang County, which shared the same evolutionary branch with O. tsutsugamushi isolated from wild mouse liver or spleen, indicating that the wild mouse plays an important role in transmitting the disease. In contrast to the sporadic cases in the past, our study is the first to disclose an epidemic and the corresponding clinical characteristics of ST in Guizhou province, which is of great significance for the prevention and treatment of regional illnesses. Author summary: In this study, 13 cases of ST were identified through clinical and laboratory diagnosis in Rongjiang County, which is the first outbreak of the disease in Guizhou province, China. It is worth noting that ST was caused by the Karp strain, which can be transmitted by wild mice, and the infected population generally had a clear history of outdoor activity. In addition to the typical clinical symptoms such as fever, chills and pneumonia, nearly half of patients with ST also exhibited abnormal electrocardiogram activity. In conclusion, our study presents a method to confirm ST through the combination of clinical and laboratory diagnosis, which provides a reference for the development of effective strategies and measures to tackle ST, and has important public health significance. [ABSTRACT FROM AUTHOR]

Simultaneous use of domestic spaces by humans and wildlife is little understood, despite global ubiquity, and can create an interface for human exposure to wildlife pathogens. Bats are a pervasive synanthropic taxon and are associated with several pathogens that can spill over and cause disease in humans. Urbanization has destroyed much natural bat habitat and, in response, many species increasingly use buildings as roosts. The purpose of this study was to characterize human interactions with bats in shared buildings to assess potential for human exposure to and spillover of bat-borne pathogens. We surveyed 102 people living and working in buildings used as bat roosts in Taita-Taveta county, Kenya between 2021 and 2023. We characterized and quantified the duration, intensity, and frequency of human-bat interactions occurring in this common domestic setting. Survey respondents reported living with bats in buildings year-round, with cohabitation occurring consistently for at least 10 years in 38% of cases. Human contact with bats occurred primarily through direct and indirect routes, including exposure to excrement (90% of respondents), and direct touching of bats (39% of respondents). Indirect contacts most often occurred daily, and direct contacts most often occurred yearly. Domestic animal consumption of bats was also reported (16% of respondents). We demonstrate that shared building use by bats and humans in rural Kenya leads to prolonged, frequent, and sometimes intense interactions between bats and humans, consistent with interfaces that can facilitate exposure to bat pathogens and subsequent spillover. Identifying and understanding the settings and practices that may lead to zoonotic pathogen spillover is of great global importance for developing countermeasures, and this study establishes bat roosts in buildings as such a setting. Author summary: Many bat species share space with people and domestic animals. Bat use of buildings, which serve as roosts where they raise young and sleep, is increasing globally with the loss of natural habitat. In such settings, interactions between bats, people, and domestic animals may occur. This may be cause for concern as bats can carry pathogens that infect people or domestic animals and interactions with bats may create opportunities for exposure to these pathogens. Until now, the characteristics of human-bat interactions in buildings, and their potential for exposing humans and domestic animals to bat-borne parasites, were unknown. In our study, people living and working in buildings used simultaneously by bats reported frequent interactions with bats and their feces, which can facilitate human exposure to bat pathogens. These interactions happened frequently and over many years. We demonstrate that shared building use by bats and people in rural Kenya leads to prolonged, frequent, and sometimes intense interactions between bats, humans, and domestic animals, consistent with exposure opportunities that can lead to pathogen spillover. Identifying the settings that may lead to human contact with pathogens is critical for developing countermeasures to mitigate public health hazards. [ABSTRACT FROM AUTHOR]

Scabies is a neglected tropical disease (NTD) with high prevalence rate in resource-limited settings. Though street children are susceptible because of lack of sanitation and contact with vectors, few attempts have been made to identify the lived experience of street children with scabies in the global south. This study explored perceived susceptibility to scabies and related lived experiences of street children in Addis Ababa. Using in-depth interviews, we collected qualitative data from selected children of the street to identify their understanding of the causes of scabies, their experiences of managing the condition, and their health-seeking behavior. Informants were recruited to include maximum variation in terms of age, sex, and experience of infestation. The study showed that scabies was common among street children and that the infestation has physical, psychological and social impacts. Study participants believed that scabies had its origin in their living conditions (including poor environmental sanitation and lack of personal hygiene), with lice playing a significant role as vectors of transmission. The informants reported visiting modern healthcare facilities, traditional healers and self-care in response to infestation. By uncovering the embodied experience of a stigmatized skin NTD in a neglected community in the global south, this study contributes to combating neglect and addressing health disparities. Having identified living conditions as the major factor contributing to susceptibility, efforts need to be exerted to change street children's living situations and other structural conditions through reunification with their families or other communities, reintegration and other exit strategies. Author summary: Evidence indicates that children living in difficult situations are at high risk of scabies infestation. Street children are one of these groups with higher susceptibility to scabies due to their living conditions. Living as a street child in resource-limited areas substantially increases vulnerability to the condition because of lack of access to healthcare services and other resources to maintain personal hygiene. This study employed a qualitative approach to understand the susceptibility of street children in Addis Ababa to scabies and their health seeking behavior. The study revealed that scabies is very common among children of the street in the area. The participants associated susceptibility to their living conditions that include sleeping arrangements, sharing of clothes and other materials. They also reported the use of both traditional and modern medicine to treat scabies. The physical, psychological, and social consequences of scabies infestation have also been identified. Although most believe that scabies relates to poor hygiene practices and attempts to prevent the condition have been reported, lack of resources posed significant challenges. We recommend sustainable reunification and reintegration of the street children to help reduce the burden of scabies among street children. [ABSTRACT FROM AUTHOR]

Background: Epigenetic modifications, such as DNA methylation and histone modifications, are pivotal in regulating gene expression pathways related to inflammation and cancer. While there is substantial research on epigenetic marks in cholangiocarcinoma (CCA), Opisthorchis viverrini-induced cholangiocarcinoma (Ov-CCA) is overlooked as a neglected tropical disease (NTD) with limited representation in the literature. Considering the distinct etiological agent, pathogenic mechanisms, and pathological manifestations, epigenetic research plays a pivotal role in uncovering markers and potential targets related to the cancer-promoting and morbidity-inducing liver fluke parasite prevalent in the Great Mekong Subregion (GMS). Emerging studies highlight a predominant hypermethylation phenotype in Opisthorchis viverrini (O. viverrini) tumor tissues, underscoring the significance of abnormal DNA methylation and histone modifications in genes and their promoters as reliable targets for Ov-CCA. Principal findings: Relevant published literature was identified by searching major electronic databases using targeted search queries. This process retrieved a total of 81 peer-reviewed research articles deemed eligible for inclusion, as they partially or fully met the pre-defined selection criteria. These eligible articles underwent a qualitative synthesis and were included in the systematic review. Within these, 11 studies specifically explored Ov-CCA tissues to investigate potential epigenetic biomarkers and therapeutic targets. This subset of 11 articles provided a foundation for exploring the applications of epigenetics-based therapies and biomarkers for Ov-CCA. These articles delved into various epigenetic modifications, including DNA methylation and histone modifications, and examined genes with aberrant epigenetic changes linked to deregulated signalling pathways in Ov-CCA progression. Conclusions: This review identified epigenetic changes and Wnt/β-catenin pathway deregulation as key drivers in Ov-CCA pathogenesis. Promoter hypermethylation of specific genes suggests potential diagnostic biomarkers and dysregulation of Wnt/β-catenin-modulating genes contributes to pathway activation in Ov-CCA progression. Reversible epigenetic changes offer opportunities for dynamic disease monitoring and targeted interventions. Therefore, this study underscores the importance of these epigenetic modifications in Ov-CCA development, suggesting novel therapeutic targets within disrupted signalling networks. However, additional validation is crucial for translating these novel insights into clinically applicable strategies, enhancing personalised Ov-CCA management approaches. Author summary: This review examines the role of epigenetic changes and the Wnt/β-catenin signalling pathway in Opisthorchis viverrini-induced cholangiocarcinoma (Ov-CCA). Through an exhaustive analysis of relevant studies, the primary epigenetic alteration identified in Ov-CCA pathogenesis is hypermethylation of tumor suppressor gene promoters. This epigenetic dysregulation shows several affected genes involved in the Wnt/β-catenin signalling pathway, which plays a central role in opisthorchiasis progression. Network analysis revealed an interaction between these genes and β-catenin, suggesting Wnt/β-catenin pathway dysregulation contributes to Ov-CCA development and progression. Most methylated genes act as tumor suppressors by normally inhibiting the Wnt/β-catenin pathway. Targeting this pathway offers therapeutic potential for Ov-CCA. Epigenetic markers hold promise as biomarkers, emphasizing the therapeutic potential of targeting disrupted gene and signalling networks through epigenetic modulators. This approach presents an innovative strategy for developing personalized therapies tailored to the molecular profiles of Ov-CCA patients. However, translating these findings into clinical practice requires rigorous validation and further in-depth exploration. Nonetheless, the insights obtained highlight the crucial necessity for ongoing research on epigenetic-based interventions, intending to enhance both diagnostic accuracy and therapeutic effectiveness in managing Ov-CCA. [ABSTRACT FROM AUTHOR]

Introduction: Brucellosis, a zoonotic infectious disease caused by bacteria of the genus Brucella, remains a significant global health concern in many parts of the world. Traditional diagnostic methods, including serological tests, suffer from limitations, including low sensibility and high false-positive rates, emphasizing the need for improved diagnostic strategies. In this study, we aimed to optimize diagnostic accuracy by reevaluating serological tests and exploring novel diagnostic algorithms. Methods: A retrospective observational study was conducted using sera collected between June 2012 and June 2023 at the French National Reference Center for Brucella. Various serological tests, including Rose Bengal plate test (RBT), standard agglutination test (SAT), Brucellacapt, and ELISA for IgM and IgG, were performed. Different diagnostic algorithms were evaluated, combining RBT with SAT, Brucellacapt, and ELISA to enhance the performance of diagnostic tests. Results: Among 3587 sera analyzed, 148 were confirmed cases of human brucellosis. Individual serological tests exhibited good sensitivity and specificity but lacked diagnostic accuracy. However, combining RBT with SAT or Brucellacapt significantly improved diagnostic performance, with reduced false positives. The most promising results were observed when an algorithm was built combining RBT, Brucellacapt, and ELISA for IgM and IgG (a score value of 0.5 with 90.5% for sensitivity, 99.7% for specificity, 92.4% for PPV, and 99.6% for NPV). Conclusions: Serological tests remain crucial for brucellosis diagnosis, but their limitations necessitate innovative diagnostic approaches. Combining multiple serological tests in diagnostic algorithms shows promise in improving diagnostic accuracy. Efforts to refine diagnostic, strengthen surveillance, and raise awareness are essential for effective brucellosis control, particularly in resource-limited settings. Author summary: Brucellosis is a zoonotic infectious disease caused by bacteria of the genus Brucella. This disease remains a significant global health concern in many parts of the world. Traditional diagnostic methods include serological tests. However, these tests have major limitations due to their low sensibility and high false-positive rates. This poses a significant challenge in clinical practice, emphasizing the necessity for careful interpretation of test results and the development of more specific diagnostic tools. In this study, we assess the utility of traditional and easily accessible serological tests. Instead of using them individually, we decided to integrate the results of these multiple testing methods. We highlight the value of combining serological results into an algorithm to enhance the diagnosis of brucellosis and achieve greater overall accuracy in diagnosis. Efforts to streamline diagnostic, bolster surveillance, and increase awareness are essential for controlling brucellosis, particularly in resource-limited settings with a high disease burden. [ABSTRACT FROM AUTHOR]

Over the past decades, the number of arthropod-borne virus (arbovirus) outbreaks has increased worldwide. Knowledge regarding the sylvatic cycle (i.e., non-human hosts/environment) of arboviruses is limited, particularly in Africa, and the main hosts for virus maintenance are unknown. Previous studies have shown the presence of antibodies against certain arboviruses (i.e., chikungunya-, dengue-, and zika virus) in African non-human primates and bats. We hypothesize that small mammals, specifically rodents, may function as amplifying hosts in anthropogenic environments. The detection of RNA of most arboviruses is complicated by the viruses' short viremic period within their hosts. An alternative to determine arbovirus hosts is by detecting antibodies, which can persist several months. Therefore, we developed a high-throughput multiplex immunoassay to detect antibodies against 15 medically relevant arboviruses. We used this assay to assess approximately 1,300 blood samples of the multimammate mouse, Mastomys natalensis from Tanzania. In 24% of the samples, we detected antibodies against at least one of the tested arboviruses, with high seroprevalences of antibodies reacting against dengue virus serotype one (7.6%) and two (8.4%), and chikungunya virus (6%). Seroprevalence was higher in females and increased with age, which could be explained by inherent immunity and behavioral differences between sexes, and the increased chance of exposure to an arbovirus with age. We evaluated whether antibodies against multiple arboviruses co-occur more often than randomly and found that this may be true for some members of the Flaviviridae and Togaviridae. In conclusion, the development of an assay against a wide diversity of medically relevant arboviruses enabled the analysis of a large sample collection of one of the most abundant African small mammals. Our findings highlight that Mastomys natalensis is involved in the transmission cycle of multiple arboviruses and provide a solid foundation to better understand the role of this ubiquitous rodent in arbovirus outbreaks. Author summary: One of the main causes of zoonotic related human morbidity and mortality is the transmission of arthropod-borne viruses such as dengue virus, Yellow Fever virus, and chikungunya virus. These viruses cannot only infect humans but also livestock, pets, and wildlife, though our understanding of their non-human hosts remains limited. Rodents are thought to be an important host for these viruses because they can be abundant, often live near humans, and some are already known to be viral hosts. However, research has focused mostly on non-human primates, neglecting other potential wild hosts. To address this gap, we have developed a high-throughput antibody test to screen rodent blood against 15 different arboviruses. Our findings reveal that Mastomys natalensis, a common African rodent species, carries antibodies that (cross-)react against these viruses. We hypothesize that immunologically naïve juveniles may drive transmission, particularly during population outbreaks. These outbreaks coincide with environmental conditions that are favorable for mosquitoes, thus increasing the risk of spillover to humans, livestock, and wildlife. Understanding the role of rodents in arbovirus transmission dynamics is crucial for mitigating zoonotic disease risks. [ABSTRACT FROM AUTHOR]

Background: Loiasis is a disease caused by the nematode Loa loa. Serious adverse events sometimes occur in people with heavy L. loa microfilaremia after ivermectin treatment. In regions of Central Africa where loiasis is endemic, this significantly impedes global elimination programs for lymphatic filariasis and onchocerciasis that use mass distribution of ivermectin. Improved diagnostic tests to identify individuals at increased risk of serious adverse events could facilitate efforts to eliminate lymphatic filariasis and onchocerciasis in this region. Methods and findings: We previously identified the L. loa protein Ll-Bhp-1 in loiasis patient sera. Here, we further characterize Ll-Bhp-1 and report development of an antigen capture ELISA to detect this antigen. This assay detected Ll-Bhp-1 in 74 of 116 (63.8%) loiasis patient sera. Ll-Bhp-1 levels were significantly correlated with L. loa microfilarial counts, and the sensitivity of the assay was highest for samples from people with high counts, (94% and 100% in people with ≥20,000 and ≥50,000 microfilaria per milliliter of blood, respectively). The antigen was not detected in 112 sera from people with other filarial infections, or in 34 control sera from the USA. Conclusions: This Ll-Bhp-1 antigen assay is specific for loiasis, and highly sensitive for identifying people with high L. loa microfilarial counts who are at increased risk for serious adverse events after ivermectin treatment. L. loa antigen detection has the potential to facilitate loiasis mapping efforts and programs to eliminate lymphatic filariasis and onchocerciasis in Central Africa. Author summary: Lymphatic filariasis and onchocerciasis are major neglected tropical diseases that have been targeted for elimination by the World Health Organization. Elimination campaigns for these infections using mass drug administration have had a huge impact in Africa. However, they face a major challenge in 11 countries in Central Africa where there is co-endemic loiasis, another filarial infection caused by the parasite Loa loa. The anti-filarial medication ivermectin that is used in elimination programs can cause serious and fatal adverse drug events in people with heavy loiasis infections, or high blood microfilarial counts. These programs would benefit from practical and affordable screening tests to detect people at high risk of adverse events from ivermectin. Furthermore, loiasis itself causes a variety of negative health outcomes and improved diagnostic tests would help control this infection. Here, we report the development of a sandwich immunoassay that detects the L. loa protein Ll-Bhp-1 in human serum samples. This assay is specific for L. loa infection and is highly sensitive in people with high microfilarial counts. Further work is needed to refine this prototype test and to determine whether antigen testing for loiasis can facilitate filarial eliminations efforts in Central Africa. [ABSTRACT FROM AUTHOR]

The formamidase (FMD) enzyme plays an important role in fungal thriving by releasing a secondary nitrogen source as a product of its activity. In Paracoccidioides species, previous studies have demonstrated the upregulation of this enzyme in a wide range of starvation and infective-like conditions. However, Paracoccidioides lutzii formamidase has not yet been defined as a virulence factor. Here, by employing in vivo infections using an fmd-silenced strain in Galleria mellonella larvae model, we demonstrate the influence of formamidase in P. lutzii's immune stimulation and pathogenicity. The formamidase silencing resulted in improper arrangement of the nodules, poor melanogenesis and decreased fungal burden. Thus, we suggest that formamidase may be a piece composing the process of molecular recognition by Galleria immune cells. Furthermore, formamidase silencing doubled the observed survival rate of the larvae, demonstrating its importance in fungal virulence in vivo. Therefore, our findings indicate that formamidase contributes to Galleria's immune incitement and establishes the role of this enzyme as a P. lutzii virulence factor. Author summary: The study of fungal neglected pathogens is of significant importance for the elucidation of the mechanisms underlying the diseases they cause. The Paracoccidioides lutzii fungus is a pathogen endemic to Latin America, which causes disease in vulnerable portions of society. To cause the disease, the fungal cells must overcome the immune system and obtain nutrients that are typically withdrawn by the host. In this regard, different molecules are crucial for the establishment of infection. The formamidase enzyme plays a role in the survival of the fungus P. lutzii. Our study, presented below, demonstrates that formamidase silencing affects the immune response of G. mellonella larvae to the fungus, resulting in impairment of nodule formation, diminished melanin production, and a reduction in fungal burden. Furthermore, silencing formamidase expression in P. lutzii doubles the survival rate of larvae infected by the fungus, showing its importance for these yeast cells to infect this animals. [ABSTRACT FROM AUTHOR]

Background: Indian subcontinent being an important region in the fight to eliminate cholera needs better cholera surveillance. Current methods miss most infections, skewing disease burden estimates. Triangulating serosurvey data, clinical cases, and risk factors could reveal India's true cholera risk. Methods: We synthesized data from a nationally representative serosurvey, outbreak reports and risk factors like water, sanitation and the Multidimensional Poverty Index, to create a composite vulnerability index for assessing state-wise cholera risk in India. We tested 7,882 stored sera samples collected during 2017–18 from individuals aged 9–45 years, for vibriocidal antibodies to Vibrio cholerae O1 using a cut-off titre ≥320 defining as elevated titre. We also extracted data from the 2015–19 Integrated Disease Surveillance Programme and published cholera reports. Results: Overall, 11.7% (CI: 10.4–13.3%) of the sampled population had an elevated titre of cholera vibriocidal antibodies (≥320). The Southern region experienced the highest incidence (16.8%, CI: 12.1–22.8), followed by the West (13.2%, CI: 10.0–17.3) and North (10.7%, CI: 9.3–12.3). Proportion of samples with an elevated vibriocidal titre (≥320) was significantly higher among individuals aged 18–45 years (13.0% CI: 11.2–15.1) compared to children 9–17 years (8.6%, CI 7.3–10.0, p<0.05); we found no differences between sex or urbanicity. Between 2015–2019, the Integrated Disease Surveillance Program (IDSP) reported 29,400 cases of cholera across the country. Using the composite vulnerability index, we found Karnataka, Madhya Pradesh, and West Bengal were the most vulnerable states in India in terms of risk of cholera. Conclusion: The present study showed that cholera infection is present in all five regions across India. The states with high cholera vulnerability could be prioritized for targeted prevention interventions. Author summary: Cholera remains a significant public health threat in areas with inadequate water, sanitation, and hygiene (WASH). The World Health Organization's Global Task Force on Cholera Control (GTFCC) launched "Ending Cholera: A Global Roadmap to 2030" to reduce cholera deaths and eliminate the disease in 20 countries by 2030 through targeted interventions, including improved WASH practices and disease surveillance. cholera surveillance, especially in developing countries seldom poses difficulty in terms of the need for laboratory infrastructure, and other resource constraints. Thereby, in this study, we utilized a viable approach with serosurvey data from 2017, cholera outbreak data, WASH indicators, and the Multidimensional Poverty Index (MPI) to create a composite vulnerability index for cholera risk in India. Overall, 11.7% of the sampled population had detectable levels of cholera antibodies reflecting previous infection. The Southern region of India experienced the highest incidence, followed by the West and North. the study showed that cholera infection is present in all five regions across India. Further, the vulnerability index which we built could be utilized in other settings so that effective tailored interventions could be applied in identified priority areas. [ABSTRACT FROM AUTHOR]

Background: Japanese encephalitis (JE) virus is the leading cause of vaccine-preventable encephalitis and a significant cause of disability in Asia and the western Pacific. Many countries have introduced JE vaccination programs, including several low resource countries following WHO's prioritization of JE vaccination in 2006. We sought to characterize the public health impact of JE vaccination programs. Methodology/Principal findings: JE case data and vaccination coverage rates, were requested from country health officials in 23 JE endemic countries and Chinese Taipei. Additional data were extracted from meeting presentations and published literature. JE incidence was compared before and after vaccination using a minimum three year period pre and post program introduction or expansion. Data suitable for analysis were available for 13 JE-endemic countries and Chinese Taipei, for either all age groups or for children aged under 15 years only. Five countries and Chinese Taipei introduced vaccine prior to 2006 and the all-age JE incidence was reduced by 73–100% in about 5–20 years following introduction. Six countries have introduced JE vaccine since 2006, and JE incidence in children aged younger than 15 years has been reduced by 14–79% as of 2015–2021. JE-specific data were unavailable before introduction in Thailand and Vietnam, but vaccination programs reduced acute encephalitis incidence by 80% and 74%, respectively. Even in the programs with greatest impact, it took several years to achieve their results. Conclusions/Significance: JE vaccination has greatly reduced JE in 13 JE-endemic countries and Chinese Taipei. Highest impact has been observed in countries that introduced prior to 2006, but it often took roughly two decades and substantial resources to achieve that level of success. For greatest possible impact, more recently introducing countries and funding agencies should commit to continuous improvements in delivery systems to sustain coverage after initial vaccine introduction. Author summary: While JE has been controlled by vaccination in a handful of economically developed countries endemic for JE transmission with disease reductions of 73–100%, introduction of JE vaccines has ocurred later in low and low-middle income countries (LLMIC) endemic for JE. Since 2006 focused effort has been undertaken to assist with establishment of sustained vaccination efforts against JE in LLMIC. The outcome has been growing evidence of the impact of vaccination against JE in LLMIC endemic for JE transmission with disease reductions of 14–79% among children aged under 15 years. We describe the public health impact of JE vaccination programs in both economically developed and LLMIC JE endemic countries and discuss how time and sustained effort on JE vaccination in LLMIC has high potential to reach the level of control seen in earlier decades in economically developed countries. [ABSTRACT FROM AUTHOR]

Background: About 600 million people are estimated to be infected with Strongyloides stercoralis, the species that causes most of the human strongyloidiasis cases. S. stercoralis can also infect non-human primates (NHPs), dogs and cats, rendering these animals putative sources for zoonotic human S. stercoralis infection. S. fuelleborni is normally found in old world NHPs but occasionally also infects humans, mainly in Africa. Dogs in southeast Asia carry at least two types of Strongyloides, only one of which appears to be shared with humans ("dog only" and "human and dog" types). For S. stercoralis with molecular taxonomic information, there is a strong sampling bias towards southeast and east Asia and Australia. Methodology/Principle findings: In order to extend the geographic range of sampling, we collected human and dog derived Strongyloides spp. and hookworms from two locations in Bangladesh and subjected them to molecular taxonomic and genomic analysis based on nuclear and mitochondrial sequences. All hookworms found were Necator americanus. Contrary to earlier studies in Asia, we noticed a rather high incidence of S. fuelleborni in humans. Also in this study, we found the two types of S. stercoralis and no indication for genetic isolation from the southeast Asian populations. However, we found one genomically "dog only" type S. stercoralis in a human sample and we found two worms in a dog sample that had a nuclear genome of the "dog only" but a mitochondrial genome of the "human and dog" type. Conclusions/Significance: S. fuelleborni may play a more prominent role as a human parasite in certain places in Asia than previously thought. The introgression of a mitochondria haplotype into the "dog only" population suggests that rare interbreeding between the two S. stercoralis types does occur and that exchange of genetic properties, for example a drug resistance, between the two types is conceivable. Author summary: More than 600 million people are infected with the nematode intestinal parasite Strongyloides stercoralis. Dogs can also carry S. stercoralis. In southeast Asia different genetic types that either infect only dogs or humans and dogs were described. Strongyloides fuelleborni, (normally found in old-world monkeys) can also infect humans, mainly in Africa. We collected Strongyloides spp. and hook worms, from humans and a dog in Bangladesh and analyzed their nuclear and mitochondrial genomes. All hookworms were Necator americanus, one of the two major human hookworm species. Contrary to the general believe that human infections with S. fuelleborni are extremely rare in Asia, we found multiple such cases, suggesting that S. fuelleborni plays a more important role as a human parasite than previously thought also in Asia. We found the two expected genetic types of S. stercoralis. For the first time we found a genomically "dog only" type worm in a person and we found two worms with nuclear genomes of the "dog only" type but mitochondrial genomes of the "human and dog" type. This suggest that rare interbreeding between the two types occurs, such that exchange of genetic properties, such as a drug resistance, between the two types is conceivable. [ABSTRACT FROM AUTHOR]

West Nile virus (WNV) is the leading mosquito-borne disease causing-pathogen in the United States. Concerningly, there are no prophylactics or drug treatments for WNV and public health programs rely heavily on vector control efforts to lessen disease incidence. Insecticides can be effective in reducing vector numbers if implemented strategically, but can diminish in efficacy and promote insecticide resistance otherwise. Vector control programs which employ mass-fogging applications of insecticides, often conduct these methods during the late-night hours, when diel temperatures are coldest, and without a-priori knowledge on daily mosquito activity patterns. This study's aims were to 1) quantify the effect of temperature on the toxicity of two conventional insecticides used in fogging applications (malathion and deltamethrin) to Culex tarsalis, an important WNV vector, and 2) quantify the time of host-seeking of Cx. tarsalis and other local mosquito species in Maricopa County, Arizona. The temperature-toxicity relationship of insecticides was assessed using the WHO tube bioassay, and adult Cx. tarsalis, collected as larvae, were exposed to three different insecticide doses at three temperature regimes (15, 25, and 35°C; 80% RH). Time of host-seeking was assessed using collection bottle rotators with encephalitis vector survey traps baited with dry ice, first at 3h intervals during a full day, followed by 1h intervals during the night-time. Malathion became less toxic at cooler temperatures at all doses, while deltamethrin was less toxic at cooler temperatures at the low dose. Regarding time of host-seeking, Cx. tarsalis, Aedes vexans, and Culex quinquefasciatus were the most abundant vectors captured. During the 3-hour interval surveillance over a full day, Cx. tarsalis were most-active during post-midnight biting (00:00–6:00), accounting for 69.0% of all Cx. tarsalis, while pre-midnight biting (18:00–24:00) accounted for 30.0% of Cx. tarsalis. During the 1-hour interval surveillance overnight, Cx. tarsalis were most-active during pre-midnight hours (18:00–24:00), accounting for 50.2% of Cx. tarsalis captures, while post-midnight biting (00:00–6:00) accounted for 49.8% of Cx. tarsalis. Our results suggest that programs employing large-scale applications of insecticidal fogging should consider temperature-toxicity relationships coupled with time of host-seeking data to maximize the efficacy of vector control interventions in reducing mosquito-borne disease burden. Author summary: Mosquito-borne pathogens such as West Nile virus (WNV) are expected to increase due to climate change, and public health programs rely on insecticides to reduce mosquito vector populations. Insecticidal fogging is often conducted during post-midnight hours when temperatures are coldest. Temperature has been shown to impact the toxicity of insecticides, but this effect is understudied, particularly for WNV vectors. The researchers aimed to test the effect of temperature on the toxicity of two common insecticides to Culex tarsalis, a primary mosquito vector of WNV, and to identify when local vectors were actively seeking a host to bite. The research location was a productive riparian (wetland) area adjacent to many urban centers in Arizona. The researchers found that both insecticides became less toxic at colder temperatures and that early evening accounted for a significant portion of captures, yet is outside of normal fogging hours. Human outdoor activities also increase during these hours, which may result in elevated risk of disease transmission. This research serves to improve our understanding and usage of insecticides to reduce disease burden. As insecticides are used globally, the findings of this research may relate to all mosquito-borne diseases, including malaria, and are relevant for agricultural pest management. [ABSTRACT FROM AUTHOR]

Background: In Victoria, Australia, children with Pacific Islander ('Pacific') ethnicities are overrepresented in acute rheumatic fever (ARF) and rheumatic heart disease (RHD). In June 2023, ARF and RHD became notifiable in Victoria. To inform public health and clinical practice, we described young Pacific patients' and their caregivers' understandings and experiences of ARF/RHD, and identified possible ways to improve the delivery of clinical care. Methods: We established a project reference group including local Pacific people to guide this research. Pacific patients who attended an ARF/RHD clinic at The Royal Children's Hospital, Melbourne, were invited to participate, as were their caregivers. A Samoan researcher conducted qualitative 'talanoa' (conversational) interviews with patients and caregivers. A second researcher conducted semi-structured interviews with treating clinicians and other stakeholders. Interview transcripts underwent thematic analysis guided by the Tuilaepa Youth Mentoring Services Pacific Youth Wellbeing Framework. Results: We interviewed 27 participants. This included nine patients and nine caregivers, all of whom were Samoan. These 18 participants expressed a desire to learn more about ARF/RHD and connect with other affected people. While some shared their experiences of having well-liked and trusted healthcare providers, patients often struggled to have two-way clinical conversations. The need to support clinicians working with high-risk populations to improve their awareness of ARF was identified. Receiving treatment on time was a top priority for affected families, despite injection pain, inconvenience and financial costs. The need to support continuity of care for young adult patients was raised by participants. Conclusions: Pacific people living with ARF/RHD and their families require additional support to receive high quality management in Victoria. Introducing a patient register and a specialist RHD nurse would enhance access to treatment, as would removing cost barriers, improving clinical awareness of ARF/RHD and creating Victoria-specific patient resources. Author summary: Acute rheumatic fever (ARF) and its serious complication rheumatic heart disease (RHD) are key markers of social injustice. In Victoria, Australia, ARF and RHD became notifiable in mid-2022. This change was instigated in response to recent work showing an ongoing burden of disease in young Victorians, particularly people of Pacific Islander ('Pacific') ethnicities. Until recently, nothing has been documented of the experiences that patients and their families have had when seeking clinical care for ARF/RHD in Victoria. We undertook this qualitative study to address this knowledge gap. Our study was co-designed by a project reference group which included local Pacific people. We used traditional Pacific 'talanoa' conversational style interviews to report on patients' and their caregivers' experiences of receiving clinical care. Our findings provide critical guidance for best practice public health management in areas such as Victoria with a low but severely ethnically inequitable disease burden. We make recommendations to support clinicians in areas with at-risk populations to enhance their awareness of ARF and RHD, and improve the delivery of clinical care. Our recommendations provide direction for future initiatives to prevent and control these serious health conditions, and better support affected people. [ABSTRACT FROM AUTHOR]

Background: Paracoccidioidomycosis (PCM) and histoplasmosis are endemic fungal diseases in South America. Both can lead to lung involvement with fungal dissemination progressing to systemic and severe clinical manifestations, especially in immunosuppressed hosts. As the population of immunosuppressed individuals has been rising, a higher occurrence of fungal infections is predicted in this setting. This poses challenges regarding the differential diagnosis due to overlapping clinical and laboratorial findings, hampering the management of cases. Objectives: In this study, the authors discuss the occurrence of a false-positive Histoplasma urinary antigen detection in a kidney transplant recipient with acute PCM. Given the scarce information about this subject, a review on literature data is provided. Methods: A comprehensive literature search was conducted to investigate previous studies that found cross-reactivity between Histoplasma urinary antigen assays in human patients with confirmed diagnosis of PCM. Additionally, an update of PCM in transplant recipients is provided. Findings: The included studies reported 120 samples from patients with PCM tested for Histoplasma antigen, presenting an overall cross-reactivity of 51.67% and 17 cases of PCM in transplant recipients. CONCLUSIONS: The galactomannan urinary antigen developed to diagnose histoplasmosis can cross react with PCM, which may represent a concern in countries where both mycoses overlap. Author summary: Paracoccidioidomycosis (PCM) and histoplasmosis are severe fungal diseases occurring simultaneously in South America. They can cause similar clinical manifestations, especially in immunosuppressed hosts. This poses challenges regarding the differential diagnosis due to overlapping clinical and laboratorial findings, hampering the management of individual cases. In this study, the authors report the occurrence of a false-positive urinary test to detect histoplasmosis in a kidney transplant recipient with acute PCM. Given the scarce information about this subject, a review on literature data is provided. As conclusion, this study highlights the urinary test developed to diagnose histoplasmosis can cross react with PCM, which may represent a concern in countries where both mycoses overlap. [ABSTRACT FROM AUTHOR]

Hemorrhagic fever with renal syndrome caused by hantaviruses has long been a serious public health issue in Yunnan Province. Hantaviruses exhibit a high extent of biodiversity in their natural hosts, particularly in mammalian hosts. This study was conducted to screen for hantaviruses in bats and rodents in Yunnan Province and elucidate their genetic characteristics and possible zoonotic disease risk. Hantaviruses were detected in 202 bats and 372 rodents with the positive rates 27.49% and 1.25% respectively. A novel lineage (named Lineage 10) of the Seoul virus (SEOV) from rodents and the geographic clustering of hantavirus in bats were identified using phylogenetic analyses of the full-length M- and S-segments. Our study suggest a high cross-species transmissibility of hantaviruses in bats and existence of a new lineage of SEOV in rodents differing significantly from other SEOVs. These results provide data to support the prevention and control of hantavirus-associated diseases in Yunnan Province. Author summary: Hantavirus is an important pathogen that can cause hemorrhagic fever with renal syndrome (HFRS) and hantavirus cardiopulmonary syndrome (HCPS). HFRS has long been a serious public health problem in Yunnan Province. However, the molecular epidemiology and host characteristics of hantavirus in Yunnan Province are still unclear. Therefore, we screened hantaviruses in bats and rodents in selected areas of Yunnan from 2018 to 2022 and found the 10th SEOV lineage in rodents, as well as new geographic clusters of Xuan son virus (XSV) and novel hantavirus in bats. We proposed the possibility of the 10th SEOV lineage in rodents as an ancestral lineage and explained the related host-switching events in bat hantaviruses. These results provide data to reveal a greater risk of zoonotic disease and support the prevention and control of hantavirus-associated diseases and in Yunnan Province. [ABSTRACT FROM AUTHOR]

Background: Coxiella burnetii, the causative agent of Q fever, and Rift Valley fever virus are two under-researched zoonotic pathogens in Ethiopia. Potential outbreaks of these diseases, in light of the high dependency of nomadic pastoralists on their livestock, poses a risk to both human and animal health in addition to risking the pastoralists livelihoods. Our study aimed to determine the seroprevalence and associated risk factors for Q fever and Rift Valley fever in pastoral communities in the Afar region of north-eastern Ethiopia. Methodology/Principal findings: This cross-sectional study screened pastoralists (n = 323) and their livestock (n = 1377) for IgG antibodies to Coxiella burnetii and Rift Valley fever virus. A seroprevalence for Q fever of 25.0% (95%CI 18.6–32.6) was found in pastoralists and 34.3% (95%CI 27.9–41.3) in livestock overall; with 51.9% in goats (95%CI 44.9–58.8), 39.9% in sheep (95%CI 24.6–51.2), 16.3% in camels (95%CI 10.4–24.6) and 8.8% in cattle (95%CI 5.0–15.0). For Rift Valley fever the seroprevalence in pastoralists was 6.1% (95%CI 3.3–11.0) and 3.9% (95%CI 2.6–5.7) in livestock overall; cattle had the highest seroprevalence (8.3%, 95%CI 3.3–19.2), followed by goats (2.7%; 95%CI 1.4–5.1), sheep (2.5%; 95%CI 1.0–5.9) and camels (1.8%; 95%CI 0.4–6.9). Human Q fever seropositivity was found to be associated with goat abortions (OR = 2.11, 95%CI 1.18–3.78, p = 0.011), while Rift Valley fever seropositivity in livestock was found to be associated with cattle abortions (OR = 2.52, 95%CI 1.05–6.08, p = 0.039). Conclusions/Significance: This study provides evidence for a notable exposure to both Q fever and Rift Valley fever in pastoralists and livestock in Afar. The outbreak potential of these pathogens warrants ongoing integrated human and animal surveillance requiring close collaboration of the human and animal health sectors with community representatives following a One Health approach. Author summary: Q fever and Rift Valley fever are two diseases that can affect both humans and animals, causing illness and death. These two diseases can cause large-scale outbreaks, not only affecting the health of individuals and communities but also resulting in a substantial economic loss at the individual, community, regional and national levels through losses in livestock products. We conducted a sero-epidemiological study in the Afar region of north-eastern Ethiopia to determine the presence of these two diseases in nomadic pastoralists and their livestock. Our results indicate that 25.0% of the pastoralists and 34.3% of their animals have had previous exposure to Q fever and 6.1% of the pastoralists and 3.9% of their animals have been exposed to Rift Valley fever. Goats appear to be the most common livestock species associated with Q fever in this region, while cattle are the most common species associated with Rift Valley fever. Our findings highlight the importance of continuous surveillance; additionally, we would recommend the development of rapid response plans for potential outbreaks that integrate the human and animal health sectors. [ABSTRACT FROM AUTHOR]

Melioidosis, a severe bacterial illness caused by Burkholderia pseudomallei, is prevalent in most parts of Thailand, including its southern region situated within the Malay Peninsula. Despite a lower reported incidence rate of melioidosis in the South compared to the Northeast, the mortality rate remains persistently high. This study aimed to better understand the epidemiology and investigate the presence of B. pseudomallei in the natural environment of southern Thailand. Using multi-locus sequence typing (MLST), we characterized B. pseudomallei isolates derived from human cases and compared them with previously reported sequence types (STs) from the same region. A total of 263 clinical isolates retrieved from 156 melioidosis patients between 2014 and 2020 were analyzed, revealing 72 distinct STs, with 25 (35%) matching STs from Finkelstein's environmental isolates collected in southern Thailand during 1964–1967. Notably, strains bearing STs 288, 84, 54, 289, and 46 were frequently found among patients. Additionally, we observed strain diversity with multiple STs in 13 of 59 patients, indicating exposure to various B. pseudomallei genotypes in the environmental sources of the infection. Environmental surveys were conducted in Songkhla Province to detect B. pseudomallei in soil and water samples where local patients lived. Of the 2737 soil samples from 208 locations and 244 water samples from diverse sources, 52 (25%) soil sampling locations and 63 (26%) water sources were cultured positive for B. pseudomallei. Positive soil samples were predominantly found in animal farming area and non-agricultural zones like mountains and grasslands, while water samples were frequently positive in waterfalls, streams, and surface runoffs, with only 9% of rice paddies testing positive. Collectively, a significant proportion of recent melioidosis cases in Songkhla Province can be attributed to known B. pseudomallei STs persisting in the environment for at least the past six decades. Further characterization of B. pseudomallei isolates from recent environment surveys is warranted. These findings illuminate the contemporary landscape of B. pseudomallei infections and their environmental prevalence in southern Thailand, contributing to the regional threat assessment in Thailand and Southeast Asia. Author summary: This study conducted a genetic analysis of Burkholderia pseudomallei, the bacterium causing a deadly tropical disease, from 156 patients in southern Thailand during 2014–2020. It found various genotypes, some of which were consistent with the genotypes found in the environment dating back to the 1960s. Extensive testing of soil and water in the region revealed contamination with B. pseudomallei. Areas such as animal farms, mountains, grasslands, and natural water sources e.g., streams and waterfalls, were mostly affected. This finding confirmed that some B. pseudomallei genotypes have had a long-standing presence in the environment, contributing to recent human cases. This study not only enhance our understanding of the spread of melioidosis throughout Southeast Asia but also highlights the importance of ongoing research and monitoring of B. pseudomallei in natural environments to address potential risks associated with the bacterium's persistence. [ABSTRACT FROM AUTHOR]

Background: This study compared the clinical sensitivity and the time-to-result of an individual testing (IT) and a cascaded pooled testing approach (CPT; a positive test result in a pooled sample triggers examination of smaller-sized pools or individual samples) for assessing the prevalence and the intensity of Schistosoma haematobium infection. We also compared the sensitivity of the CPT in detecting S. haematobium infection when deploying urine filtration microscopy (UFM) vs. urine reagent strips (URS), and testing 10 mL vs. 15 mL of urine. Methodology/Principal findings: Between October 2021 and April 2022, S. haematobium eggs were counted in urine samples collected from school-aged children living in the Afar and Gambella Regional States of Ethiopia. Urine samples were collected at baseline (n = 1,288), and one month after administration of praziquantel (n = 118). All urine samples were processed through both an IT and a CPT approach (pools of 5, 10, 20, and 40 individual samples), deploying UFM (10 mL) and URS (10 mL). In addition, 15 mL urine was processed through the CPT deploying UFM. At baseline, the prevalence of S. haematobium infection estimated when using UFM and deploying a CPT approach was significantly lower (17.3%) compared to an IT approach (31.5%). The clinical sensitivity of the CPT in detecting S. haematobium eggs was 51.7%. The sensitivity increased significantly as a function of increasing log transformed urine egg counts (UECs) of the individual samples (OR 2.71, 95%CI 1.63 ― 4.52). The sensitivity was comparable when the amount of urine examined was 10 mL (51.7%) vs. 15 ml (50.8%), and when UFM was used for testing vs. URS (51.5%). The mean log UECs estimated following the CPT approach was lower compared to the estimate by the IT (p <0.001). UECs of the individual samples estimated using the IT and CPT approaches were moderately correlated (r = 0.59 when 10 mL and 15 mL urine was examined after pooling). CPT reduced the time needed for processing urine samples and testing for S. haematobium infection by 29% with UFM and by 27.7% with URS. Conclusions/Significance: CPT based on UFM and URS techniques may help to rapidly identify areas with higher prevalence of S. haematobium infection (hotspots) in a population. However, the performance of this approach in estimating the prevalence of infection may be compromised, particularly in endemic areas with low intensity infection. Author summary: We examined the sensitivity and the time-to-result of a cascaded pooled testing approach for detecting and estimating the intensity of Schistosoma haematobium infection. Urine samples collected from school-aged children in the Afar and Gambella regions of Ethiopia were analyzed individually and in pools using both urine filtration microcopy (UFM) and urine reagent strips (URS). The cascaded pooled testing approach detected close to 52% of urine samples with S. haematobium eggs. The sensitivity increased as the egg count of the individual samples increased but was comparable when the amount of urine examined was 10 mL vs. 15 mL, and when the diagnostic test used was UFM vs. URS. Pooled testing reduced the time needed for processing urine samples and counting S. haematobium eggs by 29%. Pooled testing based on UFM may help rapidly identify areas with higher prevalence of S. haematobium infection in a population. [ABSTRACT FROM AUTHOR]

Background: Cutaneous leishmaniasis is a widespread parasitic infection in Ethiopia. Few studies have been conducted on knowledge, attitudes, and prevention practice related to cutaneous leishmaniasis, and the existing studies have been mainly without qualitative support. This study aimed to assess the knowledge, attitude, prevention practice and lived experience towards cutaneous leishmaniasis and associated factors among residents of the Kutaber district, Northeast Ethiopia. Methods: A convergent parallel mixed method was conducted among 636 residents (for quantitative) and 20 in-depth interview participants (for qualitative) of the Kutaber district from July 1 to August 15, 2022. Quantitative data were entered into Epi-Data version 4.6 and exported to SPSS version 25 for cleaning and analysis. ATLAS.ti software version 8.0 was used for the analysis of qualitative data. Results: The survey respondents in Kutaber district showed good knowledge (47.5%), a positive attitude towards (54.1%) and a good prevention practice (35.3%) regarding cutaneous leishmaniasis. Residents who were unable to read and write [AOR = 0.15] had lower odds to have good knowledge about cutaneous leishmaniasis. Residents aged >54.5 years [AOR = 0.33] had lower odds to have a positive attitude towards cutaneous leishmaniasis. Males [AOR = 1.76] had a good prevention practice towards cutaneous leishmaniasis. Five main study themes were formed following the qualitative analysis of the data. Conclusion: Residents of Kutaber district have a poor overall knowledge and prevention practice towards cutaneous leishmaniasis, despite having a positive attitude. The educational status of residents and years of residence were factors significantly associated with knowledge about cutaneous leishmaniasis. Age and years of residence were factors significantly associated with attitude towards cutaneous leishmaniasis. Gender, age, and household wealth were factors significantly associated with prevention practice towards cutaneous leishmaniasis. Cutaneous leishmaniasis in Kutaber is a true health problem. Author summary: Cutaneous leishmaniasis (CL), a prevalent disease caused by an intracellular protozoan transmitted by a vector, is a serious public health issue, particularly in Kutaber district, where it causes significant physical and psychosocial problems among the residents. Even though it is not fatal, cutaneous leishmaniasis can lead to complications, lifelong scar and disfigurement, resulting in social isolation and discrimination. We showed that this is strongly associated with the district residents' poor knowledge and prevention practice towards the disease. Cutaneous leishmaniasis, a true health problem of the community, is worsened due to limited efforts of healthcare professionals, limited roles played by the media in health information communication, budget problem, low attention given from the government, lack of information dissemination, and lack of awareness. This implied that there is a need of rigorous provision of health education on the cause, transmission mechanism, intermediate host, breeding site, sandfly vector, treatment and implementation of prevention strategies. [ABSTRACT FROM AUTHOR]

Background: In clinical practice guidelines, there is no consensus about the medications that should be initially offered to patients with brucellosis. To provide informative evidence, we compared and ranked brucellosis medications based on their efficacy and safety. Methods: For this systematic review and network meta-analysis, we searched 4 English databases and 3 Chinese databases, from the date of database inception to December 13, 2023. We included randomized controlled trials (RCTs) involving children and adolescents with brucellosis, comparing different antibiotic regimens. We excluded studies explicitly targeting patients with spondylitis brucellosis, endocarditis brucellosis, and neuro-brucellosis. The primary outcomes were overall failure (efficacy) and side effects (safety). Secondary outcomes were relapse and therapeutic failure. Pairwise meta-analysis was first examined. Data were analyzed using random effects network meta-analysis, with subgroup and sensitivity analyses performed. The Confidence in Network Meta-Analysis (CINeMA) framework was used to assess the certainty of evidence. The protocol was preregistered in PROSPERO (CRD42023491331). Results: Of the 11,747 records identified through the database search, 43 RCTs were included in the network meta-analysis. Compared with standard therapy (Doxycycline + Rifampicin), Rifampicin + Tetracyclines (RR 4.96; 95% CI 1.47 to 16.70; very low certainty of evidence), Doxycycline + TMP/SMX (RR 0.18; 95% CI 0.06 to 0.52; low certainty of evidence), Doxycycline + Quinolones (RR 0.27; 95% CI 0.11 to 0.71; low certainty of evidence), Streptomycin + Tetracyclines (RR 0.04; 95% CI 0.01 to 0.16; low certainty of evidence), and Single (RR 0.05; 95% CI 0.02 to 0.16; moderate certainty of evidence) were less efficacious. Doxycycline + Gentamicin ranked the best in efficacy (SUCRA values: 0.94), the second is Triple (SUCRA values: 0.87), and the third is Doxycycline + Streptomycin (SUCRA values: 0.78). Conclusions: Brucellosis medications differ in efficacy and safety. Doxycycline + Gentamicin, Triple, and Doxycycline + Streptomycin have superior efficacy and safety. Treatment of brucellosis should strike a balance between efficacy, safety, and cost. Author summary: Brucellosis, with nearly 500,000 new cases outbreak in more than 170 countries or regions worldwide each year, is a zoonotic infectious disease caused by Brucella. Due to the lack of an effective human vaccine and the impracticality of eradicating brucellosis in animals globally, treatment remains the primary strategy for addressing human brucellosis. However, there is no consensus in clinical practice guidelines regarding the initial medications for brucellosis treatment. Therefore, we compared and ranked brucellosis medications based on their efficacy and safety to aid clinicians in matching risk/benefits of drugs to brucellosis treatment. In this systematic review and network meta-analysis of 43 randomized clinical trials with 4,283 patients, 6 weeks of doxycycline plus 1 to 2 weeks of gentamicin or plus 2 to 3 weeks of streptomycin is the best therapy for human brucellosis. Six weeks of rifampicin plus quinolones treatment is considered as an alternative therapy. If finances allow, consider triple therapy. Single, Streptomycin + Tetracyclines, Streptomycin + TMP/SMX, and Rifampicin + TMP/SMX therapies cannot currently be recommended. [ABSTRACT FROM AUTHOR]

Mycetoma profoundly affects marginalised communities, especially in impoverished and remote areas with limited access to healthcare. This chronic and debilitating inflammatory disease highlights the typical issues of neglected tropical diseases (NTDs), such as insufficient attention, funding, and resources, which perpetuate neglect and suffering. Patients often delay seeking medical help, leading to advanced disease stages, severe complications, and lasting disabilities. The lack of medical infrastructure and skilled healthcare professionals worsens the situation, causing delays in diagnosis and inadequate treatment. Engaging affected communities in tailored interventions is essential to tackle these challenges, promote collaboration, raise awareness, and mobilise resources to improve healthcare access and enhance diagnostic and treatment capabilities. Since 1991, the Mycetoma Research Center (MRC) at the University of Khartoum, Sudan, has led community engagement initiatives aimed at improving the quality of life for mycetoma-affected individuals through education, advocacy, and local collaboration. In this communication, the MRC shares its extensive experience in community engagement to benefit mycetoma-affected communities. [ABSTRACT FROM AUTHOR]

Background: Snakebite envenoming (SBE) is a potentially life-threatening event that can lead to severe physical, mental, and economic hardships, particularly in under-resourced regions like sub-Saharan Africa. In Rwanda, there have been no epidemiological assessments of SBE to guide the Ministry of Health in its efforts to reduce the burden. This study had two main objectives: first, to estimate the incidence of snakebites across districts, and second, to describe formal versus informal healthcare seeking behaviors among snakebite victims in Eastern Province, Rwanda in 2020. Methodology: This cross-sectional study utilized a cluster sampling approach, involving Community Health Workers (CHWs) who recorded snakebite cases across seven districts. The descriptive analysis considered sampling weights, and healthcare seeking behavior was assessed based on the type of care sought as the first point of treatment. Findings: The study surveyed 390,546 individuals across 763 villages and estimated a provincial annual incidence rate of 4.3 cases per 1,000 individuals. Incidence estimates ranged from 1.1 cases per 1,000 in Nyagatare to 9.1 cases per 1,000 individuals in Bugesera and Ngoma districts. Among the 2,545 cases recorded by CHWs, three resulted in deaths. Regarding healthcare-seeking behavior, 13% of snakebite victims (143 out of 1,098) initially consulted formal care providers (CHWs, health post/center, or hospital), while 87% sought informal care (family/friends, pharmacist, or traditional healer). Approximately half of the victims (583, 53.1%) reported severe symptoms. Unsafe practices included skin cutting/burning, tourniquet application, use of black stones, and venom extraction; only 24 cases (2.2%) received anti-venom. Conclusions: This large-scale community-based assessment highlights variations in snakebite incidence between districts and confirms frequent involvement of traditional healers in management. Improving access to anti-venom and community education on the risks of ineffective practices, along with timely use of formal healthcare, are crucial. Collaboration between healthcare providers, traditional healers, community leaders, and policymakers is essential to implement targeted interventions for enhancing snakebite prevention and management strategies. Author summary: Snakebites can be life-threatening, especially in places like sub-Saharan Africa. In Rwanda, there haven't been any studies to understand how many snakebites happen and where they happen. This study wanted to find out two main things: first, how many snakebites happened in different parts of the Eastern Province of Rwanda in 2020, and second, where people who got bitten by snakes went for help—to doctors or traditional healers. The study involved 390,546 people across 763 villages. It found that each year, about 4.3 out of 1,000 people in the province get bitten by snakes, but the numbers vary in different areas. Surprisingly, only 13% of the victims initially consulted formal healthcare providers like community health workers, health centers, or hospitals. In contrast, a larger group, 87%, sought informal care from family, friends, pharmacists, or traditional healers first. About half of the people bitten by snakes experienced severe symptoms. Unfortunately, many people used unsafe methods to treat snakebites, like cutting or burning the skin, tying a tight band around the bitten area, using black stones, or trying to suck out the venom. Only a very small number of cases, just 24 out of all 1,098 victims, received anti-venom. The study suggests we need to make sure anti-venom is easier to get and teach communities about the risks of unsafe practices and why it's important to get formal healthcare quickly. It's important for doctors, traditional healers, community leaders, and government officials to work together to make sure people get safe and appropriate treatment and learn how to prevent snakebites. [ABSTRACT FROM AUTHOR]

Introduction: Scabies undermines quality of life through its highly disturbing disease symptoms, by distorting self-perception, and secondary to social stigma. Knowledge of its effect on quality of life in general and on specific aspects of day-to-day life is key to addressing the health needs of individual patients and to evaluating gains from community-based disease control interventions. Objectives: To measure the effect of scabies on the quality of life of people with the infestation. Methods: A community-based cross-sectional study was conducted in a scabies outbreak-affected district in north-western Ethiopia. The study involved 381 households and 86 adults with scabies. We used the ten-item Cardiff Dermatology Life Quality Index (DLQI) tool to collect data. Cronbach's alpha value was used to determine the internal consistency of the Amharic version of the scale. Overall and Dermatology Life Quality (DLQ) domain specific mean scores were calculated. The association between sociodemographic characteristics and scabies-related life quality impairment was tested using Kruskal-Wallis test. Results: Scabies moderately affected the quality of life of adults with scabies. The overall mean DLQI (mDLQI) score was 9.2 (SD = 7.6). 'Symptoms and feelings' and 'daily activity' DLQ domains had the highest mDLQI scores (3.5, SD = 1.9 and 2.2, SD = 2.5, respectively). 'Leisure activities' was the least affected domain 0.8 (SD = 1.1). In terms of severity, scabies had moderate or severe effect on DLQ of 54.7% of the participants and extremely severe effect was reported among 27% of the participants. However, no association was observed between sociodemographic characteristics and quality of life impairment. Conclusion: Quality of life was moderately impaired among people affected by scabies. Refocusing attention on management of disease symptoms, using standard scabies treatment, and providing psychosocial support to improve self-perception of people affected with scabies may help reduce quality of life impairment. Author summary: Scabies is a parasitic infestation of the outermost layer of the skin. Disease symptoms such as papular rash (mainly involving the crevices of the body), severe generalized itch and stigma attached to the infestation are believed to impair the quality of life of scabies patients. Despite the need for data, there is limited evidence about the relationship between scabies and quality of life. This study presents novel data on quality of life impairment associated with scabies and which aspects of patients' day-to-day lives are the most affected. This study was conducted in an outbreak-affected area of north-western Ethiopia where scabies had a moderate impact on the quality of life of the affected people. Disease symptoms and their psychosocial impact contributed the most to quality of life impairment. Participants in this study came from different sociodemographic backgrounds. Nonetheless, there were no major differences in DLQI scores by background characteristics. [ABSTRACT FROM AUTHOR]

The skin is an anatomical reservoir for African trypanosomes, yet the prevalence of extravascular parasite carriage in the population at risk of gambiense Human African Trypanosomiasis (gHAT) remains unclear. Here, we conducted a prospective observational cohort study in the HAT foci of Forecariah and Boffa, Republic of Guinea. Of the 18,916 subjects serologically screened for gHAT, 96 were enrolled into our study. At enrolment and follow-up visits, participants underwent a dermatological examination and had blood samples and superficial skin snip biopsies taken for examination by molecular and immuno-histological methods. In seropositive individuals, dermatological symptoms were significantly more frequent as compared to seronegative controls. Trypanosoma brucei DNA was detected in the blood of 67% of confirmed cases (22/33) and 9% of unconfirmed seropositive individuals (3/32). However, parasites were detected in the extravascular dermis of up to 71% of confirmed cases (25/35) and 41% of unconfirmed seropositive individuals (13/32) by PCR and/or immuno-histochemistry. Six to twelve months after treatment, trypanosome detection in the skin dropped to 17% of confirmed cases (5/30), whereas up to 25% of unconfirmed, hence untreated, seropositive individuals (4/16) were still found positive. Dermal trypanosomes were observed in subjects from both transmission foci, however, the occurrence of pruritus and the PCR positivity rates were significantly higher in unconfirmed seropositive individuals in Forecariah. The lower sensitivity of superficial skin snip biopsies appeared critical for detecting trypanosomes in the basal dermis. These results are discussed in the context of the planned elimination of gHAT. Author summary: The skin is a reservoir for African trypanosomes. Here, we conducted a prospective study in Forecariah and Boffa, Guinea, to estimate the proportion of skin-dwelling parasites in the population. Of the 18,916 subjects screened for HAT, 96 were enrolled into our study. Participants underwent a dermatological examination and had blood samples and superficial skin biopsies taken for examination by molecular and immuno-histological methods. In individuals seropositive for HAT, dermatological symptoms were significantly more frequent. Trypanosome DNA was detected in the blood of 67% of confirmed cases and 9% of unconfirmed seropositive individuals. However, parasites were detected in the skin of up to 71% of confirmed cases and 41% of unconfirmed seropositive individuals. After treatment, trypanosome detection in the skin dropped to 17% of confirmed cases, whereas up to 25% of unconfirmed, hence untreated, seropositive individuals were still found positive. Dermal trypanosomes were observed in subjects from both regions; however, the occurrence of itching and the PCR positivity were significantly higher in unconfirmed seropositive individuals in Forecariah. The lower sensitivity of superficial skin biopsies appeared critical for detecting trypanosomes. These results are discussed in the context of the planned elimination of HAT. [ABSTRACT FROM AUTHOR]

Background: Loiasis is a disease of relevance in endemic populations and there has been advocacy for its inclusion on the World Health Organization's neglected tropical diseases list. As loiasis-related healthcare-seeking behaviors and related costs are unknown, we aimed to evaluate these aspects in a population residing in an endemic region in Gabon. Methods: Data were collected during a community-based, cross-sectional study assessing the disease burden due to loiasis. Diagnostics for microfilaremia were performed and a history of eyeworm was obtained. In addition, a standardized questionnaire about type of healthcare resources and frequency of use, as well as respective associated costs was administered to each participant. Loiasis related healthcare-seeking behaviors were evaluated, and the associated monetary burden was estimated as a secondary outcome of the study. Findings: Individuals diagnosed with loiasis more frequently reported any healthcare-seeking (OR 1.52 (95%CI: 1.21–1.91)), self-medicating (OR 1.62 (1.26–2.08)), inability to work (OR 1.86 (1.47–2.35)), and consulting with traditional healers (logOdds 1.03 (0.52–1.53)), compared to loiasis negative individuals. The most frequently reported treatment for the eyeworm was traditional herbs. The estimated healthcare associated costs, per positive individual, was US-$ 58 (95% CI: 21–101) per year, which would correspond to 3.5% of the reported mean household income. Extrapolation to the rural population of Gabon (n = 204,000), resulted in an annual monetary burden estimate of US-$ 3,206,000 (1,150,000–5,577,000). Interpretation: Loiasis patients have demonstrated healthcare needs, often consulted traditional healers, and used traditional treatments for disease specific symptoms. Further, loiasis seems to be associated with substantial direct and indirect costs for individuals and thus may cause a relevant economic burden for endemic populations and economies of affected countries. Author summary: Loiasis has long been considered a negligible infection, however disease perception is changing as more data on significant disease-associated morbidity and mortality are being published. Still, many important aspects of the disease, such as the socioeconomic impact, are unknown. This study was part of a larger project on loiasis associated disease burden and aimed to evaluate healthcare seeking behaviors and estimate associated costs in individuals with loiasis living in a highly endemic region. We found that loiasis is associated with healthcare consultation, self-medicating, and sick leave. Self-medicating for treatment of disease specific symptoms was frequent. Traditional treatments and pain medication were most frequently used. Based on these data, we estimated loiasis related direct healthcare costs and indirect costs due to sickness, resulting in a monetary burden of US-$ 58 (95% CI: 21–101) per loiasis positive individual per year. Extrapolating these numbers to the rural population of Gabon (n = 204,000) would correspond to a monetary burden of US-$ 3,206,000 (95% CI: 1,150,000–5,577,000). Thus, loiasis is not only associated with morbidity and mortality but also with healthcare-seeking and may cause a relevant socioeconomic burden on affected populations. Therefore, considering loiasis a negligible disease is not appropriate anymore. [ABSTRACT FROM AUTHOR]

Rift Valley fever (RVF) is an important zoonotic viral disease affecting several species of domestic and wild ruminants, causing major economic losses and dozens of human deaths in various geographical areas of Africa, where it is endemic. Although it is not present in Europe, there is a risk of its introduction and spread linked to globalisation and climate change. At present, the only measure that could help to prevent the disease is vaccination of flocks in areas at risk of RVF. Available live attenuated vaccines are an effective means of controlling the disease, but their use is often questioned due to residual virulence, particularly in susceptible hosts such as pregnant sheep. On the other hand, no vaccine is currently licensed for use in humans. The development of safe and effective vaccines is therefore a major area of research. In previous studies, we selected under selective mutagenic pressure a highly attenuated RVFV 56/74 virus variant called 40Fp8. This virus showed an extremely attenuated phenotype in both wild-type and immunodeficient A129 (IFNARKO) mice, yet was still able to induce protective immunity after a single inoculation, thus supporting its use as a safe, live attenuated vaccine. To further investigate its safety, in this work we have analysed the attenuation level of 40Fp8 in immunosuppressed mice (A129) when administered by the intranasal route, and compared it with other attenuated RVF viruses that are the basis of vaccines in use or in development. Our results show that 40Fp8 has a much higher attenuated level than these other viruses and confirm its potential as a candidate for safe RVF vaccine development. Author summary: Vaccines that use attenuated viruses are highly effective in terms of providing protection, duration, breadth, and quality of the immune response. However, they may pose a risk to immunosuppressed individuals or in certain situations, such as during pregnancy. It is therefore important to analyze the residual virulence of attenuated vaccines to ensure their safety. In this work, we analysed the safety of the RVF virus vaccine variant 40Fp8 developed in our laboratory. We studied its effect on immunodeficient mice and used a more aggressive route of administration such as the intranasal delivery route. To assess the degree of attenuation of 40Fp8, we compared it with other attenuated prototype RVF vaccines. The results clearly demonstrate that 40Fp8 is highly attenuated, and its residual virulence level is extremely low compared to other vaccine viruses tested. [ABSTRACT FROM AUTHOR]

Introduction: Culex quinquefasciatus is one of the most important carriers of human pathogens. Using the insecticides is one of the most important methods of combating this vector. But the genetic resistance created in Culex quinquefasciatus led to disruption in the fight against this pest. Consequently, it is necessary to know the level of resistance to fight this vector. Based on this, the present study was conducted to investigate the prevalence of kdr resistance in Culex quinquefasciatus against organochlorine insecticides in the world. Methods: This study was conducted by systematic review, and meta-analysis on the prevalence of kdr resistance and mortality rate in Culex quinquefasciatus against organochlorine insecticides in the world. All pertinent articles were extracted and analyzed in accordance with this information during an unrestricted search of the scientific databases Web of Science, PubMed, Scopus, biooan.org, Embase, ProQuest, and Google Scholar until the end of November 2023. Statistical analysis of data was done using fixed and random effects model in meta-analysis, I2 index, Cochran's test, and meta-regression by STATA version 17 software. Results: Seventy articles were included in the meta-analysis process. Based on the findings, the prevalence of Kdr in Culex quinquefasciatus against organochlorine insecticide was estimated at 63.1%. Moreover, the mortality rate against the insecticide deltamethrin was 46%, DDT 18.5%, permethrin 42.6%, malathion 54.4% and lambdacyhalothrin 53%. Conclusion: More than half of Cx. quinquefasciatus had Kdr. This vector was relatively resistant to DDT and permethrin insecticides and sensitive to malathion, deltamethrin and lambdacyhalothrin. In order to prevent the development of resistance to alternative insecticides, it is consequently critical to combat this vector with efficacious insecticides. Author summary: Using the insecticides is one of the most important methods of combating this vector. Based on this, the present study was conducted to investigate the prevalence of kdr resistance in Culex quinquefasciatus against organochlorine insecticides in the world. This study was conducted by systematic review, and meta-analysis on the prevalence of kdr resistance and mortality rate in Culex quinquefasciatus against organochlorine insecticides in the world. Based on the findings, the prevalence of Kdr in Culex quinquefasciatus against organochlorine insecticide was estimated at 63.1%. Moreover, the mortality rate against the insecticide deltamethrin was 46%, DDT 18.5%, permethrin 42.6%, malathion 54.4% and lambdacyhalothrin 53%. More than half of Cx. quinquefasciatus had Kdr. This vector was relatively resistant to DDT and permethrin insecticides and sensitive to malathion, deltamethrin and lambdacyhalothrin. [ABSTRACT FROM AUTHOR]

Cutaneous leishmaniasis (CL) is a neglected tropical disease caused by Leishmania parasites, that can cause long-term chronic disabilities. The clinical presentation of CL varies in both type and severity. CL presents as three main clinical forms: localised lesions (localised cutaneous leishmaniasis, LCL); mucocutaneous leishmaniasis (MCL) that affects the mucosa of the nose or the mouth; or as disseminated not ulcerating nodules (diffuse cutaneous leishmaniasis, DCL). Here we recruited a cohort of CL patients in a newly established leishmaniasis treatment centre (LTC) in Lay Gayint, Northwest Ethiopia, and collected detailed demographic and clinical data. The results of our study show that more males than females present to the LTC to seek diagnosis and treatment. 70.2% of CL patients presented with LCL and 20.8% with MCL. A small number of patients presented with DCL, recidivans CL (a rare form of CL where new lesions appear on the edges of CL scars) or with a combination of different clinical presentations. The duration of illness varied from 1 month to 180 months. Over a third of CL patients had additional suspected CL cases in their household. Despite the majority of CL patients having heard about CL, only a minority knew about its transmission or that it could be treated. Most CL patients lived in areas where environmental factors known to be associated with the transmission of CL were present. This work highlights that CL is an important public health problem in Lay Gayint and emphasises the urgent need for more CL awareness campaigns, better health education and better disease management practices. Author summary: Cutaneous leishmaniasis (CL) is a neglected tropical disease caused by species of the Leishmania parasite. It causes skin lesions that can be categorised into three main clinical presentations: i) localised cutaneous leishmaniasis (LCL), with one or several ulcerated lesions; ii) mucocutaneous leishmaniasis (MCL) that affects the mucosa of the nose or the mouth; iii) disseminated CL (DCL), that is characterised by widespread nodules. While LCL usually heals spontaneously, MCL and DCL require treatment; and patients often experience relapses. CL leaves disfiguring scars, that can result in societal stigmatisation. In this study, we recruited a cohort of CL patients in a newly established leishmaniasis treatment centre (LTC) in Lay Gayint, Northwest Ethiopia, and collected detailed demographic and clinical data. We found that most CL patients presented with LCL, followed by MCL, and that more males than females present to the LTC. The duration of illness varied from 1 to 180 months. Over a third of CL patients had additional suspected CL cases in their home. Despite most patients having heard about CL, only a minority knew about its transmission or that it could be treated. This work emphasises that CL is an important public health problem in this area of Ethiopia. [ABSTRACT FROM AUTHOR]

Chagas disease, caused by the parasite Trypanosoma cruzi, affects over 6 million people, mainly in Latin America. Two different clinical phases, acute and chronic, are recognised. Currently, 2 anti-parasitic drugs are available to treat the disease (nifurtimox and benznidazole), but diagnostic methods require of a relatively complex infrastructure and trained personnel, limiting its widespread use in endemic areas, and the access of patients to treatment. New diagnostic methods, such as rapid tests (RDTs) to diagnose chronic Chagas disease, or loop-mediated isothermal amplification (LAMP), to detect acute infections, represent valuable alternatives, but the parasite's remarkable genetic diversity might make its implementation difficult. Furthermore, determining the efficacy of Chagas disease treatment is complicated, given the slow reversion of serological anti-T. cruzi antibody reactivity, which may even take decades to occur. New biomarkers to evaluate early therapeutic efficacy, as well as diagnostic tests able to detect the wide variety of circulating genotypes, are therefore, urgently required. To carry out studies that address these needs, high-quality and traceable samples from T. cruzi-infected individuals with different geographical backgrounds, along with associated clinical and epidemiological data, are necessary. This work describes the framework for the creation of such repositories, following standardised and uniform protocols, and considering the ethical, technical, and logistic aspects of the process. The manual can be adapted according to the resources of each laboratory, to guarantee that samples are obtained in a reproducible way, favouring the exchange of data among different work groups, and their generalizable evaluation and analysis. The main objective of this is to accelerate the development of new diagnostic methods and the identification of biomarkers for Chagas disease. Author summary: The diagnosis of Chagas disease requires costly equipment and trained personnel, which unavailability hinders access to diagnosis, and treatment, in vast areas of endemic regions. Additionally, timely assessment of treatment efficacy is complicated due to the slow reversion of serological anti-T. cruzi reactivity. Hence, there is an urgent need for biomarkers of early therapeutic efficacy and disease prognosis, as well as more practical diagnostic tools. To conduct studies that can address these needs, it is essential to have collections of clinical samples with good quality, traceability, and appropriately associated clinical-epidemiological information. In this work, we provide a standard protocol to collect, process, store, and transport clinical samples from Chagas disease patients. The manual was produced upon reaching a consensus among the experts within the NHEPACHA network, a coalition of clinical and academic researchers from the Americas and Spain that pursues the identification and validation of new biomarkers and diagnostics for Chagas disease. [ABSTRACT FROM AUTHOR]

There is currently no single, easy-to-use, reliable indicator to assess whether a face has been washed with soap in the context of trachoma elimination. This study aimed to compare survey report, script-based pictorial recall and facial cleanliness indicators as alternatives to structured observation for measuring face washing behaviour. This method validation study was nested in the Stronger-SAFE trial, Oromia Region, Ethiopia. Structured observation was conducted in randomly selected households for three hours from dawn. The primary caregiver in each household participated in a survey to capture (self)-reported behaviour and/or script-based pictorial recall, a routine-based diary activity to covertly capture information on face washing behaviour of themself and any children aged 1–12. Children 4–12 years old directly participated in the survey and pictorial recall in a subset of households. The facial cleanliness of children aged 1–12 was assessed qualitatively and using the quantitative Personal Hygiene Assessment Tool (qPHAT). Prevalence estimates, sensitivity, specificity and predictive values were computed for each behavioural indicator with observation data as the gold standard. The appropriateness of script-based pictorial recall was assessed using baseline and 3-month follow-up data. Baseline data were collected from 204 households in 68 clusters. Survey estimates of face washing and face washing with soap among caregivers and children were 32% to 60% and 5% to 31% higher than observed behaviour, respectively. Face washing prevalence estimates from pictorial recall were lower than survey estimates and comparable with observations for some face washing with soap indicators (0.3% to 13% higher than observations). Specificity of pictorial recall indicators was high (85% to 99%), but the sensitivity was low (0% to 67%), resulting in a low positive predictive value for all indicators. Both qualitative facial cleanliness indicators and qPHAT scores were poorly correlated with observed face washing earlier that morning. Pictorial recall overestimated face washing with soap among both caregivers and children following intervention delivery but not at baseline. Survey (self)-reported data on face washing is highly inaccurate. Script-based pictorial recall does not correctly classify those who wash their face with soap, and is subject to differential bias following intervention exposure, and facial cleanliness is a poor indicator of recent face washing in settings where faces become rapidly dirty again after washing. Alternatives to structured observation cannot be recommended to monitor the effectiveness of face washing interventions in community settings. Trial Registration ISRCTN registry ISRCTN40760473, https://doi.org/10.1186/ISRCTN40760473. Author summary: This study aimed to identify easy-to-use indicators for face washing behaviour to improve the evaluation of behaviour change interventions in the context of trachoma elimination. The study compared reported behaviour, script-based pictorial recall (a routine-based diary activity to covertly capture information on face washing behaviour) and facial cleanliness indicators with direct observation of face washing. Facial cleanliness assessment and qPHAT scores were poorly correlated with observed behaviour. Caution should be applied before using facial cleanliness metrics as a proxy for face washing. Survey report produced much higher estimates of face washing and face washing with soap than were observed. Pictorial recall overestimated face washing with soap in the intervention arm following intervention exposure, suggesting the metric reflects knowledge rather than behaviour. None of the tested alternatives to structured observation can be recommended to monitor the effectiveness of face washing interventions in community settings. [ABSTRACT FROM AUTHOR]