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Rabies is a lethal zoonotic encephalomyelitis that causes an estimated 59,000 human deaths yearly worldwide. Although developing countries of Asia and Africa bear the heaviest burden, surveillance and disease detection in these countries is often hampered by the absence of local laboratories able to diagnose rabies and/or the difficulties of sample shipment from low-access areas to national reference laboratories. Filter papers offer a convenient cost-effective alternative for the sampling, shipment, and storage of biological materials for the diagnosis of many pathogens including rabies virus, yet the properties of diagnostic tests using this support have not been evaluated thoroughly. Sensitivity and specificity of molecular diagnosis of rabies infection using a reverse transcription followed by a hemi-nested polymerase chain reaction (RT-hn-PCR) either directly on brain tissue or using brain tissue dried on filter paper were assessed on 113 suspected field animal samples in comparison to the direct fluorescent antibody test (FAT) recommended by the World Health Organization as one of the reference tests for rabies diagnosis. Impact of the duration of the storage was also evaluated. The sensitivity and the specificity of RT-hn-PCR i) on brain tissue were 96.6% (95% CI: [88.1–99.6]) and 92.7% (95% CI: [82.4–98.0]) respectively and ii) on brain tissue dried on filter paper 100% (95% CI: [93.8–100.0]) and 90.9% (95% CI: [80.0–97.0]) respectively. No loss of sensitivity of RT-hn-PCR on samples of brain tissue dried on filter paper left 7 days at ambient temperature was detected indicating that this method would enable analyzing impregnated filter papers sent to the national reference laboratory at ambient temperature within a 1-week shipment time. It could therefore be an effective alternative to facilitate storage and shipment of samples from low-access areas to enhance and expand rabies surveillance., Author summary Dogs are responsible for 99% of human rabies deaths. Facilitating diagnostic of rabid dogs can help further identify i) people who were exposed to rabies in order to encourage them to seek post exposure prophylaxis and ii) other exposed animals to break the transmission chain (i.e. prevent them from further transmitting the virus to other people and animals). Yet, the reference diagnostic method for animal rabies requires brain samples to be collected post-mortem and shipped under temperature-controlled conditions. The shipping of such samples is complex in low-access areas, especially in low income countries, which are often those that bear the heaviest rabies burden. Filter papers offer a convenient alternative for biological sample shipment and storage. Here the efficiency of a molecular diagnostic method applied to brain tissue dried on filter paper is compared to one of the reference methods, the direct fluorescent antibody test. Our results show it has an excellent sensitivity (it does not miss any positive samples), even when filter papers are left 7 days at ambient temperature. These results let us foresee a cost-effective alternative facilitating shipment, storage and testing of samples from rabies suspected animals from low-access areas. This could considerably enhance and expand rabies surveillance in low-income countries, allowing a more comprehensive evaluation of rabies burden, and thus reinforcing arguments for allocating funds to rabies control policies.

Background In 2005, Bangladesh, India and Nepal agreed to eliminate visceral leishmaniasis (VL) as a public health problem. The approach to this was through improved case detection and treatment, and controlling transmission by the sand fly vector Phlebotomus argentipes, with indoor residual spraying (IRS) of insecticide. Initially, India applied DDT with stirrup pumps for IRS, however, this did not reduce transmission. After 2015 onwards, the pyrethroid alpha-cypermethrin was applied with compression pumps, and entomological surveillance was initiated in 2016. Methods Eight sentinel sites were established in the Indian states of Bihar, Jharkhand and West Bengal. IRS coverage was monitored by household survey, quality of insecticide application was measured by HPLC, presence and abundance of the VL vector was monitored by CDC light traps, insecticide resistance was measured with WHO diagnostic assays and case incidence was determined from the VL case register KAMIS. Results Complete treatment of houses with IRS increased across all sites from 57% in 2016 to 70% of houses in 2019, rising to >80% if partial house IRS coverage is included (except West Bengal). The quality of insecticide application has improved compared to previous studies, average doses of insecticide on filters papers ranged from 1.52 times the target dose of 25mg/m2 alpha-cypermethrin in 2019 to 1.67 times in 2018. Resistance to DDT has continued to increase, but the vector was not resistant to carbamates, organophosphates or pyrethroids. The annual and seasonal abundance of P. argentipes declined between 2016 to 2019 with an overall infection rate of 0.03%. This was associated with a decline in VL incidence for the blocks represented by the sentinel sites from 1.16 per 10,000 population in 2016 to 0.51 per 10,000 in 2019. Conclusion Through effective case detection and management reducing the infection reservoirs for P. argentipes in the human population combined with IRS keeping P. argentipes abundance and infectivity low has reduced VL transmission. This combination of effective case management and vector control has now brought India within reach of the VL elimination targets., Author summary Visceral Leishmaniasis (VL), also known as kala-azar, is a major parasitic disease in South Asia (Indian subcontinent), with 85% of the disease incidence in India. Historically VL had been controlled and almost eliminated with Indoor Residual Spraying (IRS) using dichlorodiphenyltrichloroethane (DDT). However, reinitiating this approach in 2015 failed due to high insecticide resistance in the sand fly vector and poor IRS quality, meaning that VL elimination targets were not met. To improve this the National Vector Borne Disease Control Programme changed to an effective insecticide, alpha-cypermethrin and altered the mode of application to compression pumps. Sentinel sites were established to monitor the entomological indicators, these showed the positive impact of these changes from 2016 to 2019. During this period the overall incidence of disease has decreased, and India is now on track to reach it’s target incidence for VL of less than 1/1000 people at the sub-district (block) level.

Mosquito vectors lay their white eggs in the aquatic milieu. During early embryogenesis water passes freely through the transparent eggshell, which at this moment is composed of exochorion and endochorion. Within two hours the endochorion darkens via melanization but even so eggs shrink and perish if removed from moisture. However, during mid-embryogenesis, cells of the extraembryonic serosa secrete the serosal cuticle, localized right below the endochorion, becoming the third and innermost eggshell layer. Serosal cuticle formation greatly reduces water flow and allows egg survival outside the water. The degree of egg resistance to desiccation (ERD) at late embryogenesis varies among different species: Aedes aegypti, Anopheles aquasalis and Culex quinquefasciatus eggs can survive in a dry environment for ≥ 72, 24 and 5 hours, respectively. In some adult insects, darker-body individuals show greater resistance to desiccation than lighter ones. We asked if egg melanization enhances mosquito serosal cuticle-dependent ERD. Species with higher ERD at late embryogenesis exhibit more melanized eggshells. The melanization-ERD hypothesis was confirmed employing two Anopheles quadrimaculatus strains, the wild type and the mutant GORO, with a dark-brown and a golden eggshell, respectively. In all cases, serosal cuticle formation is fundamental for the establishment of an efficient ERD but egg viability outside the water is much higher in mosquitoes with darker eggshells than in those with lighter ones. The finding that pigmentation influences egg water balance is relevant to understand the evolutionary history of insect egg coloration. Since eggshell and adult cuticle pigmentation ensure insect survivorship in some cases, they should be considered regarding species fitness and novel approaches for vector or pest insects control., Author summary Mosquitoes transmit various causative agents of diseases and the blockage of vector life cycle is an effective way to hamper disease transmission. The egg is the least known life stage and understanding it can contribute with novel strategies for mosquito control. Mosquitoes lay eggs in water collections, some of which are temporary. At early embryogenesis eggs are prone to lose water, leading to dehydration and death. During embryogenesis the serosal cuticle is produced, it wraps the embryo and contributes to the egg protection, allowing it to survive outside the water. Curiously, this resistance varies among mosquitoes: Aedes, Anopheles and Culex eggs can survive outside the water for long, intermediate and short periods, respectively. Here, we show that these differences are related to the degree of eggshell melanization (melanin is a dark pigment): darker eggs resists more against water loss. We confirmed that melanin increases survival outside the water employing a mosquito mutant that does not melanize properly. The protection conferred by melanin is dependent on the formation of the serosal cuticle. Our results contribute to the study of the evolution of egg coloration in insects and we identified one of the reasons why Aedes aegypti eggs survive for several months outside water.

Background Work to control the gambiense form of human African trypanosomiasis (gHAT), or sleeping sickness, is now directed towards ending transmission of the parasite by 2030. In order to supplement gHAT case-finding and treatment, since 2011 tsetse control has been implemented using Tiny Targets in a number of gHAT foci. As this intervention is extended to new foci, it is vital to understand the costs involved. Costs have already been analysed for the foci of Arua in Uganda and Mandoul in Chad. This paper examines the costs of controlling Glossina palpalis palpalis in the focus of Bonon in Côte d’Ivoire from 2016 to 2017. Methodology/Principal findings Some 2000 targets were placed throughout the main gHAT transmission area of 130 km2 at a density of 14.9 per km2. The average annual cost was USD 0.5 per person protected, USD 31.6 per target deployed of which 12% was the cost of the target itself, or USD 471.2 per km2 protected. Broken down by activity, 54% was for deployment and maintenance of targets, 34% for tsetse surveys/monitoring and 12% for sensitising populations. Conclusions/Significance The cost of tsetse control per km2 of the gHAT focus protected in Bonon was more expensive than in Chad or Uganda, while the cost per km2 treated, that is the area where the targets were actually deployed, was cheaper. Per person protected, the Bonon cost fell between the two, with Uganda cheaper and Chad more expensive. In Bonon, targets were deployed throughout the protected area, because G. p. palpalis was present everywhere, whereas in Chad and Uganda G. fuscipes fuscipes was found only the riverine fringing vegetation. Thus, differences between gHAT foci, in terms of tsetse ecology and human geography, impact on the cost-effectiveness of tsetse control. It also demonstrates the need to take into account both the area treated and protected alongside other impact indicators, such as the cost per person protected., Author summary Sleeping sickness is a fatal disease in Africa caused by trypanosomes transmitted by the bite of infected tsetse flies. The World Health Organization has set the absence of new infections as a goal for 2030. To achieve this, screening and treatment of patients is supplemented by tsetse control. Tiny Targets are small insecticide-impregnated panels of blue cloth and black netting which attract and kill tsetse. To maintain the momentum in these last stages of eliminating the disease, it is important to understand the costs of vector control. This paper presents the results of two years’ cost monitoring in Bonon, an area covering 130 km2, home to 120,000 people. Since 2016, ~2,000 Tiny Targets have been deployed annually at a cost of USD 471 per km2 per year. Bonon is a degraded forested area and a relatively high number of targets was required as tsetse are present throughout. Nevertheless, the cost comes to only USD 0.5 per person protected per year. Compared to operations using Tiny Targets in Uganda and Chad, the cost was higher per km2 protected, reflecting the need to place targets throughout the area, but low per person protected compared to Chad, highlighting the need to use a range of comparators.

Mass drug administration (MDA), targeted at school-aged children (SAC) is recommended by the World Health Organization for the control of morbidity induced by soil-transmitted helminth (STH) infection in endemic countries. However, MDA does not prevent reinfection between treatment rounds, and research suggests that only treating SAC will not be sufficient to interrupt transmission of STH. In countries with endemic infection, such as Ethiopia, the coverage, community-groups targeted, and rates of reinfection will determine how effective MDA is in suppressing transmission in the long-term. In this paper, individually-linked longitudinal data from three epidemiological STH surveys conducted between November 2018 and November 2020 in the Wolaita region of Ethiopia are analysed to determine how STH prevalence and intensity changes according to individual level treatment data collected over two rounds of MDA. This study demonstrates that while community-wide MDA successfully reduces overall infection intensity across the villages treated, the observed levels of non-compliance to treatment by individuals acts to maintain levels of parasite abundance whereby transmission interruption is not possible at to, despite reasonable levels of MDA coverage in the communities studied (ranging from 65% to 84% of the village populations). This quantifies with substantial data the often-postulated difference between coverage (accepting treatment) and compliance (swallowing of treatment), the latter impacting the former to a previously unquantified level. The paper highlights the need to focus treatment to partially treated, or never treated groups of individuals within existing community wide MDA control activities to interrupt the transmission of STH, and to reduce the basic reproductive number, R0, of the parasites to less than unity in value.

Background Visceral leishmaniasis (VL) is targeted for elimination as a public health problem in Nepal by 2023. For nearly three decades, the core vector control intervention in Nepal has been indoor residual spraying (IRS) with pyrethroids. Considering the long-term use of pyrethroids and the possible development of resistance in the vector Phlebotomus argentipes sand flies, we monitored the susceptibility status of their field populations to the insecticides of different classes, in villages with and without IRS activities in recent years. Methodology/Principal findings Sand flies were collected from villages with and without IRS in five VL endemic districts from August 2019 to November 2020. The WHO susceptibility test procedure was adopted using filter papers impregnated at the discriminating concentrations of insecticides of the following classes: pyrethroids (alpha-cypermethrin 0.05%, deltamethrin 0.05%, and lambda-cyhalothrin 0.05%), carbamates (bendiocarb 0.1%) and organophosphates (malathion 5%). Pyrethroid resistance intensity bioassays with papers impregnated with 5× of the discriminating concentrations, piperonyl butoxide (PBO) synergist-pyrethroid bioassays, and DDT cross-resistance bioassays were also performed. In the IRS villages, the vector sand flies were resistant (mortality rate 50) with pyrethroids was observed in all villages. The pyrethroid resistance intensity was low, and the susceptibility improved at 5× of the discriminating concentrations. Enhanced pyrethroid susceptibility after pre-exposure to PBO and the DDT-pyrethroid cross-resistance were evident. Conclusions/Significance Our investigation showed that P. argentipes sand flies have emerged with pyrethroid resistance, suggesting the need to switch to alternative classes of insecticides such as organophosphates for IRS. We strongly recommend the regular and systematic monitoring of insecticide resistance in sand flies to optimize the efficiency of vector control interventions to sustain VL elimination efforts in Nepal.

The expansion of urban ecosystems and climate change, both outcomes of massive lifestyle changes, contribute to a series of side effects such as environmental deterioration, spread of diseases, increased greenhouse gas emissions and introduction of invasive species. In the case of the Athens metropolitan area, an invasive mosquito species—the Asian tiger mosquito (Aedes albopictus)–has spread widely in the last decade. This spread is favoured within urban environments and is also affected by changing climatic trends. The Asian tiger mosquito is accompanied by risks of mosquito-borne diseases, greater nuisance levels, and increased expenses incurring for its confrontation. The main aims of this paper are (i) to estimate the various costs associated with the control of this invasive species, as well as its health and nuisance impacts, (ii) to evaluate the level of citizens’ well-being from averting these impacts and (iii) to record citizens’ and experts’ perceptions regarding alternative control measures. Evidence shows that experts tend to place a high value on mosquito control when associated with serious health risks, while citizens are more sensitive and concerned about the environmental impacts of control methods. The synthesis of results produced by the current study could act as a preliminary guide for the estimation of societal welfare from the confrontation of similar problems in the context of a complex ecosystem., Author summary This paper is based on several years’ collaboration among researchers from various disciplines, key health policy makers and stakeholders in an attempt to evaluate the economic dimensions related to the presence of the Asian Tiger Mosquito (Aedes albopictus) and the challenges of tackling mosquito-borne disease outbreaks in Greece and Southern Europe. Similar studies have been conducted and continue to be published in Europe and the USA examining the socioeconomic benefit from the implementation of relevant control and prevention strategies. These studies conclude that there are significant benefits related both to the reduction of nuisance levels and the reduction of the health risks posed by various mosquito species. In our case, the application of an updated economic analysis on the effectiveness of relevant public control and prevention programs provides essential information for public health decision-making, bearing in mind the significant restructuring of the public sector and the fiscal crisis apparent in the European South.

Background We were tasked by the World Health Organization (WHO) to address the following question: What techniques should be used to diagnose Schistosoma infections in snails and in the water in potential transmission sites? Our goal was to review and evaluate the available literature and provide recommendations and insights for the development of WHO’s Guidelines Development Group for schistosomiasis control and elimination. Methodology We searched several databases using strings of search terms, searched bibliographies of pertinent papers, and contacted investigators who have made contributions to this field. Our search covered from 1970 to Sept 2020. All papers were considered in a PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) framework, and retained papers were grouped by technique and subjected to our GRADE (Grading of Recommendations, Assessment, Development and Evaluations) evidence assessment profile determined in consultation with WHO. We also considered issues of sensitivity, specificity, coverage, cost, robustness, support needs, schistosome species discrimination, and relevant detection limits. Principal findings Our PRISMA process began with the perusal of 949 articles, of which 158 were retained for data extraction and evaluation. We identified 25 different techniques and for each applied a GRADE assessment considering limitations, inconsistency, imprecision, indirectness, and publication bias. We also provide advantages and disadvantages for each category of techniques. Conclusions Our GRADE analysis returned an assessment of moderate quality of evidence for environmental DNA (eDNA), qPCR and LAMP (Loop-mediated isothermal amplification). No single ideal diagnostic approach has yet been developed, but considerable recent progress has been made. We note a growing trend to use eDNA techniques to permit more efficient and replicable sampling. qPCR-based protocols for follow-up detection offer a versatile, mature, sensitive, and specific platform for diagnosis though centralized facilities will be required to favor standardization. Droplet digital PCR (ddPCR) can play a complementary role if inhibitors are a concern, or more sensitivity or quantification is needed. Snail collection, followed by shedding, is encouraged to provide specimens for sequence verifications of snails or schistosomes. LAMP or other isothermal detection techniques offer the prospect of less expensive and more distributed network of analysis but may face standardization and verification challenges related to actual sequences amplified. Ability to detect schistosome infections in snails or in the water is needed if control and elimination programs hope to succeed. Any diagnostic techniques used need to be regularly verified by the acquisition of DNA sequences to confirm that the detected targets are of the expected species. Further improvements may be necessary to identify the ideal schistosome or snail sequences to target for amplification. More field testing and standardization will be essential for long-term success., Author summary Global efforts are underway to reach the goal of elimination of schistosomiasis as a public health problem by 2030. A crucial step in elimination programs is the verification of elimination, including surveillance of former transmission foci. This systematic review assessed and evaluated a wide range of diagnostic tools for detection of Schistosoma parasites in snails and water. Our analysis revealed that along with standard snail shedding methods, molecular methods such as PCR, qPCR and LAMP are becoming the widely adopted standard approaches to detect schistosomes in snails. Recent developments in eDNA methods are further enabling novel surveillance capabilities for snails and schistosomes in water and are likely to become more widely adopted. While there is currently a plethora of techniques to choose from, there is a clear need for further field testing and development of standardized protocol for the most promising among them, including eDNA, ddPCR, qPCR and LAMP methods. Future studies focused on field-worthy detection approaches and their efficacy and sensitivity in the field will be a corner stone in development of control and elimination programs.

Introduction A number of neglected tropical diseases are targeted for elimination or eradication. An effective surveillance system is critical to determine if these goals have been achieved and maintained. Trachoma has two related but morphologically different presentations that are monitored for elimination, the active infectious form of trachoma and trachomatous trichiasis (TT), the progression of the disease. There are a number of lessons learnt from the Guinea worm surveillance system that are particularly compatible for TT surveillance and the onchocerciasis surveillance system which can provide insights for surveillance of the infectious form of trachoma. Methods/Principal findings A literature search of peer-reviewed published papers and grey literature was conducted using PUBMED and Google Scholar for articles relating to dracunculiasis or Guinea worm, onchocerciasis and trachoma, along with surveillance or elimination or eradication. The abstracts of relevant papers were read and inclusion was determined based on specified inclusion and exclusion criteria. The credibility and bias of relevant papers were also critically assessed using published criteria. A total of 41 papers were identified that were eligible for inclusion into the review. The Guinea worm programme is designed around a surveillance-containment strategy and combines both active and passive surveillance approaches, with a focus on village-based surveillance and reporting. Although rumour reporting and a monetary incentive for the identification of confirmed Guinea worm cases have been reported as successful for identifying previously unknown transmission there is little unbiased evidence to support this conclusion. More rigorous evidence through a randomised controlled trial, influenced by motivational factors identified through formative research, would be necessary in order to consider applicability for TT case finding in an elimination setting. The onchocerciasis surveillance strategy focuses on active surveillance through sentinel surveillance of villages and breeding sites. It relies on an entomological component, monitoring infectivity rates of black flies and an epidemiological component, tracking exposure to infection in humans. Challenges have included the introduction of relatively complex diagnostics that are not readily available in onchocerciasis endemic countries and target thresholds, which are practically unattainable with current diagnostic tests. Although there is utility in monitoring for infection and serological markers in trachoma surveillance, it is important that adequate considerations are made to ensure evidence-based and achievable guidelines for their utility are put in place. Conclusions/Significance The experiences of both the Guinea worm and onchocerciasis surveillance strategies have very useful lessons for trachoma surveillance, pre- and post-validation. The use of a monetary reward for identification of TT cases and further exploration into the use of infection and serological indicators particularly in a post-validation setting to assist in identifying recrudescence would be of particular relevance. The next step would be a real-world evaluation of their relative applicability for trachoma surveillance., Author summary The design of a surveillance system needs to be carefully thought out to ensure it provides sufficient evidence to determine if a disease or infection is eliminated or eradicated. If inappropriate it can lead to on-going transmission and resurgence of infection or disease or the unnecessary continuation of interventions, wasting valuable resources. Guinea worm is a disease that is painful and debilitating, for which there is no drug or vaccine. The aim is to eradicate the disease and as such the Guinea worm programme is designed around a strategy of identification of cases and their containment to prevent onward transmission. Onchocerciasis if left untreated can lead to blindness. The aim is to eliminate the disease through the interruption of transmission. A literature review was conducted to determine available evidence and identify lessons that can be learnt from the surveillance of both diseases for the design of trachoma surveillance strategies in the endgame. The potential utility of rumour reporting and a monetary incentive for the identification of a confirmed case of Guinea worm could be explored for trichiasis case finding. Trichiasis is the progression of trachoma and leads to significant ocular morbidity. The introduction of tests for infection and antibodies and the utility of sentinel surveillance as utilised for onchocerciasis are interesting considerations for active trachoma surveillance post-validation and has potential to identify recrudescence cost-effectively. The experiences of both the Guinea worm and onchocerciasis surveillance strategies have very useful lessons that can be trialled for trachoma surveillance. However, their real-world applicability and implications for trachoma need to be evaluated before any changes in guidelines are proposed.

Domestic dogs are responsible for 99% of all cases of human rabies and thus, mass dog vaccination has been demonstrated to be the most effective approach towards the elimination of dog-mediated human rabies. Namibia demonstrated the feasibility of this approach by applying government-led strategic rabies vaccination campaigns to reduce both human and dog rabies incidences in the Northern Communal Areas of Namibia since 2016. The lessons learnt using paper-based form for data capturing and management of mass dog vaccination campaign during the pilot and roll out phase of the project (2016–2018) led to the implementation of a simple and accurate data collection tool in the second phase (2019–2022) of the rabies elimination program. In this paper, we describe the implementation of such custom-developed vaccination tracking device, i.e. the Global Alliance for Rabies Control (GARC) Data Logger (GDL), and the integration of the collected data into a website-based rabies surveillance system (Rabies Epidemiological Bulletin—REB) during 2019 and 2020 campaigns. A total of 10,037 dogs and 520 cats were vaccinated during the 2019 campaign and 13,219 dogs and 1,044 cats during the 2020 campaign. The vaccination data were recorded with the GDL and visualized via REB. Subsequent GIS-analysis using gridded population data revealed a suboptimal vaccination coverage in the great majority of grid cells (82%) with a vaccination coverage below 50%. Spatial regression analysis identified the number of schools, estimated human density, and adult dog population were associated with the vaccination performance. However, there was an inverse correlation to human densities. Nonetheless, the use of the GDL improved data capturing and monitoring capacity of the campaign, enabling the Namibian government to improve strategies for the vaccination of at-risk areas towards achieving adequate vaccination coverage which would effectively break the transmission of rabies., Author summary We used a custom-developed vaccination tracking device—the Global Alliance for Rabies Control (GARC) Data Logger—to capture dog rabies vaccination data during the 2019 and 2020 mass vaccination campaign in the Northern Communal Areas of Namibia, and then integrated the collected data into the web-based Rabies Epidemiological Bulletin, a rabies-specific disease surveillance platform for rabies-endemic countries. This approach allowed automatic collation, analysis and, visualization of data and drastically improved the data capturing and monitoring capacity of the Namibian government led campaign. Additionally, subsequent GIS analysis enabled a better estimation of vaccination coverage at a much higher spatial resolution, thus identifying areas where improvements in the vaccination strategy are needed to ensure long-term success of the project.

Background Onchocerciasis transmission across international borders is not uncommon, yet a coordinated cross border stops mass drug administration (MDA) decision has not been documented. Methods/Principle findings The Galabat-Metema focus involves neighboring districts on the border between Sudan and Ethiopia. Mass drug administration (MDA) was provided once and subsequently twice per year in this focus, with twice-per-year beginning in Ethiopia’s Metema subfocus in 2016 and in the Sudan’s Galabat subfocus in 2008. Ov16 ELISA-based serosurveys were conducted in 6072 children under 10 years of age in the Metema subfocus in 2014, and 3931 in the Galabat in 2015. Between 2014 and 2016, a total of 27,583 vector Simulium damnosum flies from Metema and 9,148 flies from Galabat were tested by pool screen PCR for Onchocerca volvulus O-150 DNA. Only 8 children were Ov16 seropositive (all in the Metema subfocus); all were negative by skin snip PCR. The upper limit of the 95% confidence interval (UCL) for Ov16 seropositive was, Author summary Onchocerciasis is a vector-borne tropical disease caused by the parasitc worm, Onchocerca volvulus. The vector is Simulium flies that breed in fast flowing rivers and streams. The infection can cause skin disease and loss of vision, giving risk to the common name of ‘river blindness’ for the condition. Ivermectin (Mectizan) mass drug administration (MDA) given at high coverage for many years can interrupt transmission such that MDA can eventually be stopped. The present paper provides highlights of onchocerciasis transmission interruption in the first known coordinated international cross border focus with ivermectin MDA program in Africa, where 99% of the global onchocerciasis burden exists. This focus known as Galabat-Metema is shared between Ethiopia and Sudan, both of which have established a nationwide policy for elimination of onchocerciasis. The paper provides the history of MDA in the border area and the results leading a coordinated end to MDA in the focus in 2017.

Through national efforts and regional cooperation under the umbrella of the Regional Program for the Elimination of Rabies, dog and human rabies have decreased significantly in Latin America and Caribbean (LAC) countries over the last three decades. To achieve this decline, LAC countries had to develop national plans, and consolidate capabilities such as regular mass dog vaccination, opportune post-exposure prophylaxis and sensitive surveillance. This paper presents longitudinal data for 21 LAC countries on dog vaccination, PEP and rabies surveillance collected from the biannual regional meeting for rabies directors from 1998–2014 and from the Regional Epidemiologic Surveillance System for Rabies (SIRVERA). Differences in human and dog rabies incidence rates and dog vaccination rates were shown between low, middle and high-income countries. At the peak, over 50 million dogs were vaccinated annually in national campaigns in the countries represented. The reported number of animal exposures remained fairly stable during the study period with an incidence rate ranging from 123 to 191 reported exposures per 100,000 people. On average, over 2 million doses of human vaccine were applied annually. In the most recent survey, only 37% of countries reported that they had sufficient financial resources to meet the program objectives. The data show a sufficient and sustained effort of the LAC countries in the area of dog vaccination and provide understanding of the baseline effort required to reduce dog-mediated rabies incidence., Author summary Globally, dog-mediated rabies kills over 55,000 people on an annual basis and disproportionally affects socio-economically disadvantaged populations. For over 4 decades, the Latin America and Caribbean (LAC) region have worked towards the elimination of dog-mediated rabies succeeding in a substantial reduction of both human and dog rabies cases. This paper summarizes the last 15 years of the national and regional rabies elimination efforts, through a series of performance indicators, including government mass dog vaccination campaigns and post-exposure prophylaxis for humans. The most important message from the tail of the rabies epidemic in the LAC Region is one of endurance by the countries and the region to achieve rabies control and elimination. The performance indicators shown in this paper are tools for the LAC region to evaluate its progress and refine its approach, and for other regions to understand the scope of the effort required to reach elimination.

In recent years, an increased focus has been placed upon the possibility of the elimination of soil-transmitted helminth (STH) transmission using various interventions including mass drug administration. The primary diagnostic tool recommended by the WHO is the detection of STH eggs in stool using the Kato-Katz (KK) method. However, detecting infected individuals using this method becomes increasingly difficult as the intensity of infection decreases. Newer techniques, such as qPCR, have been shown to have greater sensitivity than KK, especially at low prevalence. However, the impact of using qPCR on elimination thresholds is yet to be investigated. In this paper, we aim to quantify how the sensitivity of these two diagnostic tools affects the optimal prevalence threshold at which to declare the interruption of transmission with a defined level of confidence. A stochastic, individual-based STH transmission model was used in this study to simulate the transmission dynamics of Ascaris and hookworm. Data from a Kenyan deworming study were used to parameterize the diagnostic model which was based on egg detection probabilities. The positive and negative predictive values (PPV and NPV) were calculated to assess the quality of any given threshold, with the optimal threshold value taken to be that at which both were maximised. The threshold prevalence of infection values for declaring elimination of Ascaris transmission were 6% and 12% for KK and qPCR respectively. For hookworm, these threshold values are lower at 0.5% and 2% respectively. Diagnostic tests with greater sensitivity are becoming increasingly important as we approach the elimination of STH transmission in some regions of the world. For declaring the elimination of transmission, using qPCR to diagnose STH infection results in the definition of a higher prevalence, than when KK is used., Author summary Soil-transmitted helminths are categorised as a neglected tropical disease and comprise four dominant species (two hookworms, Trichuris trichuria & Ascaris lumbricoides) that affect the poorest people in the world. The World Health Organisation (WHO) has made great strides in reducing the morbidity induced by STH infections in pre-school aged and school-aged children through mass drug administration. Many countries are now considering moving from morbidity reduction in school-aged children to community-wide treatment with the aim of transmission elimination. These helminths reproduce sexually within a human host and therefore both male and female worms must be present to produce fertilized eggs. The density of female and male worms below which mating success is too low to sustain parasite populations is defined as the ‘breakpoint’ in transmission. Both the prevalence and intensity of infection are very low as this breakpoint is approached when worm numbers are highly aggregated in their distribution within the human host population. Consequently, it becomes increasingly challenging to identify infected individuals using standard microscopic diagnostic tools (such as Kato-Katz). New and more sensitive molecular diagnostics tools, such as qPCR, are a necessity in settings where communities are moving towards the interruption of transmission. This paper demonstrates that the threshold to declare interruption of transmission is 50% lower when microscopic techniques are applied compared with molecular techniques.

Background The decade following the Regional Strategic Framework for Visceral Leishmaniasis (VL) elimination in 2005 has shown compelling progress in the reduction of VL burden in the Indian subcontinent. The Special Programme for Research and Training in Tropical Diseases (TDR), hosted by the World Health Organization (WHO) and other stakeholders, has coordinated and financed research for the development of new innovative tools and strategies to support the regional VL elimination initiative. This paper describes the process of the TDR’s engagement and contribution to this initiative. Methodology/principal findings Multiple databases were searched to identify 152 scientific papers and reports with WHO funding or authorship affiliation around the following 3 framework strategies: detection of new cases, morbidity reduction, and prevention of infection. TDR has played a critical role in the evaluation and subsequent use of the 39-aminoacid–recombinant kinesin antigen (rK39) rapid diagnostic test (RDT) as a confirmatory test for VL in the national program. TDR has supported the clinical research and development of miltefosine and single-dose liposomal amphotericin B as a first-line treatment against VL. TDR has engaged with in-country researchers, national programme managers, and partners to generate evidence-based interventions for early detection and treatment of VL patients. TDR evaluated the quality, community acceptance, and cost effectiveness of indoor residual spraying, insecticide-treated bed nets, insecticide-impregnated durable wall linings, insecticidal paint, and environmental management as tools for integrated vector management in reducing sandfly density. Conclusions/significance TDR’s engagement with country policy makers, scientists, and clinicians in the development of effective diagnosis, treatment, case detection, and vector control represents an important example of TDR’s stewardship toward the elimination of VL in the Indian subcontinent., Author summary Since the early days of the kala-azar elimination programme in the Indian subcontinent, TDR has engaged with national control and research institutions to conduct research aimed at informing country policy and practice to identify and treat cases and to prevent transmission of the infection. This includes the evaluation of the rK39 rapid diagnostic test for the diagnosis of VL, the clinical development and evaluation of first-line treatments (miltefosine and liposomal amphotericin B), the generation of evidence-based tools for early detection and complete clinical management of VL, the evaluation of the cost effectiveness of indoor residual spraying, insecticide-treated bed nets, insecticide-impregnated durable wall linings, and environmental vector management as tools for integrated vector management. The interaction and interdependence between implementation research, technical advice, partnership, and policy is yet another example of TDR’s stewardship contribution and empowerment toward VL elimination in the Indian subcontinent. Continuing investment in translational research from the bench to the bedside to public health, established jointly by national control programmes, academics, and TDR coordinators, is imperative to block transmission and prevent a resurgence of VL in the future.

Background Cutaneous leishmaniasis (CL) is a major public health problem in Libya. In this paper, we describe the eco-epidemiological parameters of CL during the armed conflict period from January 2011 till December 2012. Current spatiotemporal distributions of CL cases were explored and projected to the future using a correlative modelling approach. In addition the present results were compared with our previous data obtained for the time period 1995–2008. Methodology/Principal findings We investigated 312 CL patients who presented to the Dermatology Department at the Tripoli Central Hospital and came from 81 endemic areas distributed in 10 districts. The patients presented with typical localized lesions which appeared commonly on the face, arms and legs. Molecular identification of parasites by a PCR-RFLP approach targeting the ITS1 region of the rDNA was successful for 81 patients with two causative species identified: L. major and L. tropica comprised 59 (72.8%) and 22 (27.2%) cases, respectively. Around 77.3% of L. tropica CL and 57.7% of L. major CL caused single lesions. Five CL patients among our data set were seropositive for HIV. L. tropica was found mainly in three districts, Murqub (27.3%), Jabal al Gharbi (27.3%) and Misrata (13.7%) while L. major was found in two districts, in Jabal al Gharbi (61%) and Jafara (20.3%). Seasonal occurrence of CL cases showed that most cases (74.2%) admitted to the hospital between November and March, L. major cases from November till January (69.4%), and L. tropica cases mainly in January and February (41%). Two risk factors were identified for the two species; the presence of previously infected household members, and the presence of rodents and sandflies in patient’s neighborhoods. Spatiotemporal projections using correlative distribution models based on current case data and climatic conditions showed that coastal regions have a higher level of risk due to more favourable conditions for the transmitting vectors. Conclusion Future projection of CL until 2060 showed a trend of increasing incidence of CL in the north-western part of Libya, a spread along the coastal region and a possible emergence of new endemics in the north-eastern districts of Libya. These results should be considered for control programs to prevent the emergence of new endemic areas taking also into consideration changes in socio-economical factors such as migration, conflicts, urbanization, land use and access to health care., Author summary Cutaneous leishmaniasis (CL) is a skin infection caused by a single-celled parasite that is transmitted by the bite of a phlebotomine sandfly. CL is the most common form of leishmaniasis characterized by localized lesions in the skin and mucous membranes. The disease is prevalent in all countries around the Mediterranean Basin. In this paper, we describe spatiotemporal and eco-epidemiological parameters of CL in Libya. Moreover, we explored current spatiotemporal distributions of CL cases and explored the future projection of the disease. Our study indicates the presence of higher risk of CL in the coastal regions of Libya. Future projection until 2060 showed a trend of increasing incidence of CL in the north-western part of Libya, a spread along the coastal region and a possible emergence of new endemics in the north-eastern districts of Libya. These scenarios should be considered by health authorities in order to develop appropriate intervention strategies and plan effective control programs.

Introduction Dengue is a rapidly emerging vector-borne Neglected Tropical Disease, with a 30-fold increase in the number of cases reported since 1960. The economic cost of the illness is measured in the billions of dollars annually. Environmental change and unplanned urbanization are conspiring to raise the health and economic cost even further beyond the reach of health systems and households. The health-sector response has depended in large part on control of the Aedes aegypti and Ae. albopictus (mosquito) vectors. The cost-effectiveness of the first-ever dengue vaccine remains to be evaluated in the field. In this paper, we examine how it might affect the cost-effectiveness of sustained vector control. Methods We employ a dynamic Markov model of the effects of vector control on dengue in both vectors and humans over a 15-year period, in six countries: Brazil, Columbia, Malaysia, Mexico, the Philippines, and Thailand. We evaluate the cost (direct medical costs and control programme costs) and cost-effectiveness of sustained vector control, outbreak response and/or medical case management, in the presence of a (hypothetical) highly targeted and low cost immunization strategy using a (non-hypothetical) medium-efficacy vaccine. Results Sustained vector control using existing technologies would cost little more than outbreak response, given the associated costs of medical case management. If sustained use of existing or upcoming technologies (of similar price) reduce vector populations by 70–90%, the cost per disability-adjusted life year averted is 2013 US$ 679–1331 (best estimates) relative to no intervention. Sustained vector control could be highly cost-effective even with less effective technologies (50–70% reduction in vector populations) and in the presence of a highly targeted and low cost immunization strategy using a medium-efficacy vaccine. Discussion Economic evaluation of the first-ever dengue vaccine is ongoing. However, even under very optimistic assumptions about a highly targeted and low cost immunization strategy, our results suggest that sustained vector control will continue to play an important role in mitigating the impact of environmental change and urbanization on human health. If additional benefits for the control of other Aedes borne diseases, such as Chikungunya, yellow fever and Zika fever are taken into account, the investment case is even stronger. High-burden endemic countries should proceed to map populations to be covered by sustained vector control., Author summary Transmitted by the Aedes mosquito, dengue affects more than 100 countries and is rapidly emerging as the leading vector-borne disease. There has been a 30-fold increase in the number of cases reported since 1960. The cost of the illness to the health system and to society at large is estimated at several billions of dollars annually. The health sector response has depended in large part on controlling mosquito populations during outbreaks. Recently, the first-ever dengue vaccine received regulatory approval for use in several countries. However, its roll-out and long-term impact still needs to be evaluated in the field. In this paper, we examine how the introduction of this vaccine might alter the investment case for sustained effort to control mosquitoes. To our knowledge, this is the first economic evaluation of mosquito control in the era of the dengue vaccine. We model the cost and effects of mosquito control in Brazil, Columbia, Malaysia, Mexico, the Philippines, and Thailand. We evaluate the cost-effectiveness of mosquito control in the presence of a vaccine that does not offer full protection to all individuals. Our results suggest that sustained mosquito control will continue to be cost-effective, even if roll-out of the current vaccine is highly targeted and low-cost. These results support current global policies and strategies for the prevention and control of dengue.

The Global Program to Eliminate Lymphatic Filariasis (GPELF) has made significant progress in many countries. GPELF has delivered 5,600 million treatments to more than 763 million people living in 61 countries between the years 2000 and 2014, and it was estimated to have prevented 36 million clinical cases and saved 175 million disability adjusted life years (DALYs) [1]. Although elimination of LF is a major goal of the London Declaration on Neglected Tropical Diseases (NTDs), it is unlikely that LF will be eliminated by the target year of 2020 [2]. The key strategy of the elimination program comprises repeated annual rounds of mass drug administration (MDA) to populations at risk of acquiring the infection with diethylcarbamazine (DEC) plus albendazole outside of Africa and with ivermectin plus albendazole in Africa [3]. MDA reduces microfilariae (Mf) in human blood that are necessary for transmission by mosquito vectors, and one aim of the program is to interrupt transmission. Repeated rounds of MDA are required because current regimens fail to kill all adult worms and completely clear Mf following single-dose treatments. Successful programs have sustained annual MDA with high compliance for several years. Seventy-three countries were considered to be endemic for LF in 2015, and 55 of them still required MDA [4]. The GPELF has lagged in sub-Saharan Africa, where only 2 of 35 LF-endemic countries have stopped MDA and started post-MDA surveillance. Many countries in this region have either not started MDA or have less than 65% geographical coverage. Hence, a more effective treatment strategy could have a major impact on LF elimination in Africa. The goal of onchocerciasis intervention in Africa has also shifted in recent years from control towards elimination [5, 6]. As with LF, the key strategy for onchocerciasis intervention is MDA. In most countries, this is accomplished with annual community-directed treatment with ivermectin (CDTi). Modeling studies predict that current strategies will permit many countries to reach targets for stopping MDA for onchocerciasis by 2028, but intervention is likely to be required for a much longer period in some areas (especially in Central Africa) [7]. While annual or semiannual CDTi for 15 to 17 years eliminated the infection in some areas, it was not sufficient to stop transmission in others [8, 9]. Therefore, extensive research is currently ongoing to identify drugs that are more effective than ivermectin. Drug development studies are time and resource intensive, and it is unlikely that a novel drug for onchocerciasis will be registered to significantly advance the progress of onchocerciasis elimination within the next 5 years. In this paper, we discuss the potential value of combination therapy with three currently approved drugs (ivermectin, DEC, and albendazole, or IDA) for advancing LF and onchocerciasis elimination programs in Africa. Because DEC treatment can cause ocular adverse events in persons with heavy Onchocerca volvulus infections, studies of the new combination will have to be conducted with proper precautions. Therefore, this paper also discusses prior experience with DEC in onchocerciasis patients and a strategy for studying the effects of IDA in patients with onchocerciasis.

Background The Bangladesh Lymphatic Filariasis (LF) Elimination Programme has made significant progress in interrupting transmission through mass drug administration (MDA) and has now focussed its efforts on scaling up managing morbidity and preventing disability (MMDP) activities to deliver the minimum package of care to people affected by LF clinical conditions. This paper highlights the Bangladesh LF Programme’s success in conducting a large-scale cross-sectional survey to determine the number of people affected by lymphoedema and hydrocoele, which enabled clinical risk maps to be developed for targeted interventions across the 34 endemic districts (19 high endemic; 15 low endemic). Methodology/Principal findings In the 19 high endemic districts, 8,145 community clinic staff were trained to identify and report patients in their catchment area. In the 15 low endemic districts, a team of 10 trained field assistants conducted active case finding with cases reported via a SMS mHealth tool. Disease burden and prevalence maps were developed, with morbidity hotspots identified at sub-district level based on a combination of the highest prevalence rates per 100,000 and case-density rates per square kilometre (km2). The relationship between morbidity and baseline microfilaria (mf) prevalence was also examined. In total 43,678 cases were identified in the 19 high endemic districts; 30,616 limb lymphoedema (70.1%; female 55.3%), 12,824 hydrocoele (29.4%), and 238 breast/female genital swelling (0.5%). Rangpur Division reported the highest cases numbers and prevalence of lymphoedema (26,781 cases, 195 per 100,000) and hydrocoele (11661 cases, 169.6 per 100,000), with lymphoedema predominately affecting females (n = 21,652). Rangpur and Lalmonirhat Districts reported the highest case numbers (n = 11,199), and prevalence (569 per 100,000) respectively, with five overlapping lymphoedema and hydrocoele sub-district hotspots. In the 15 low endemic districts, 732 cases were identified; 661 lymphoedema (90.2%; female 39.6%), 56 hydrocoele (7.8%), and 15 both conditions (2.0%). Spearman’s correlation analysis found morbidity and mf prevalence significantly positively correlated (r = 0.904; p, Author summary The Global Programme to Eliminate Lymphatic Filariasis (GPELF) requires lymphatic filariasis (LF) endemic countries, such as Bangladesh, to estimate the number of lymphoedema and hydrocoele cases in order to deliver the minimum package of care required to control morbidity and reduce patient suffering. This paper highlights the Bangladesh LF Elimination Programme’s progress in training more than 8000 community health workers to identify more than 44,000 cases across 34 endemic districts where approximately 70 million people are at risk. The morbidity data collected enabled the creation of a national database and a series of risk maps of lymphoedema and hydrocoele to be developed, which highlighted the significant burden in northern Rangpur Division, especially of lymphoedema among female patients. The Bangladesh LF Elimination Programme’s efforts to identify LF cases across all endemic districts represents one of the most comprehensive national databases on LF clinical cases in the world. It provides an informative database for health workers to use in the delivery of the minimum package of care and a template for other countries to adopt and develop national strategies to manage morbidity and prevent disability as recommended by GPELF.

Background Scabies is a common dermatological condition, affecting more than 130 million people at any time. To evaluate and/or predict the effectiveness and cost-effectiveness of scabies interventions, disease transmission modelling can be used. Objective To review published scabies models and data to inform the design of a comprehensive scabies transmission modelling framework to evaluate the cost-effectiveness of scabies interventions. Methods Systematic literature search in PubMed, Medline, Embase, CINAHL, and the Cochrane Library identified scabies studies published since the year 2000. Selected papers included modelling studies and studies on the life cycle of scabies mites, patient quality of life and resource use. Reference lists of reviews were used to identify any papers missed through the search strategy. Strengths and limitations of identified scabies models were evaluated and used to design a modelling framework. Potential model inputs were identified and discussed. Findings Four scabies models were published: a Markov decision tree, two compartmental models, and an agent-based, network-dependent Monte Carlo model. None of the models specifically addressed crusted scabies, which is associated with high morbidity, mortality, and increased transmission. There is a lack of reliable, comprehensive information about scabies biology and the impact this disease has on patients and society. Discussion Clinicians and health economists working in the field of scabies are encouraged to use the current review to inform disease transmission modelling and economic evaluations on interventions against scabies., Author summary Scabies is a neglected tropical disease affecting more than 130 million people, with major costs on health care systems worldwide. While effective treatments exist, it is unknown which treatment strategies result in the best outcomes against the lowest costs, and to what extent this differs between communities. Health economic modelling can help answer these questions, but has rarely been used in this disease area. This review discusses all available scabies transmission models (n = 4), and uses them to create a new, comprehensive modelling framework. This framework can be used as aid for creating a scabies transmission model, the details of which will be determined by the context (population) and the question being addressed. The current paper also reviews the data that is needed to inform scabies modelling: on scabies biology, quality of life and resource use. Unfortunately, available data is limited and particularly data on crusted scabies (associated with high morbidity and mortality rates) is rare. With this review, we hope to assist researchers and policy makers to predict and/or evaluate the cost-effectiveness of interventions against scabies in their population(s) of interest. To tackle scabies, it is key to use effective treatment strategies in a cost-effective and sustainable way. The models and data described in this review, may help researchers, clinicians and funding bodies to facilitate this.

Background Over the last decade, implementation research and a science of global health delivery have emerged as important vehicles to improve the effectiveness of interventions. Efforts to control neglected tropical diseases (NTD) operate in challenging circumstances and with marginalized populations, making attention to context-specific details particularly relevant. Socio-anthropological insights have much to offer a science of NTD delivery. In this paper, an accessible and actionable framework for understanding NTD intervention effectiveness, based on socio-anthropological research, is presented and its utility for program planning and monitoring and evaluation is outlined. Methodology/Principal findings The framework was developed inductively by comparatively analyzing three rapid ethnographic studies undertaken in Eastern Africa (2010–2013) on three different large-scale NTD interventions: rabies elimination in Tanzania, sleeping sickness control in Uganda and the prevention of parasitic worms in Zambia. The framework includes five “intervention domains” where the effectiveness of these interventions was negotiated and determined at the local level. This involves: 1) the terrain of intervention (including seasonality and geographical variability); 2) community agency (including local knowledge, risk perceptions, behaviors, leadership and social pressure); 3) the strategies and incentives of field staff (skills, motivations, capabilities and support); 4) the socio-materiality of technology (characteristics of intervention tools and the adoption process itself); and 5) the governance of interventions (policy narratives, available expertise, bureaucracy, politics and the utilization of knowledge). The paper illustrates the importance of each of these domains by drawing on the case study research, presenting lessons learnt and practical recommendations for how such insights could improve intervention delivery. Conclusions/Significance To help close the gap between efficacy and effectiveness in NTD programs, it is important that field staff: 1) generate meaningful knowledge about contextual factors; 2) use this knowledge to tailor field strategies; and 3) create routine mechanisms to account for the dynamic process of implementation itself. The framework presented here offers a simple analytical tool to strengthen these knowledge-to-action relationships existing project planning tools, drawing on the insights of socio-anthropology., Author summary Many efficacious tools exist to control NTDs, but effectively moving these tools and approaches from the boardroom to the village is a complicated socio-political process. In the era of Sustainable Development Goals, global health has become more focused on improving the delivery of existing interventions. Greater attention to implementation research, including the value of social science perspectives, has followed in an effort to build a science of global health delivery. This paper presents an accessible and actionable socio-anthropological framework for understanding the effectiveness factors of NTD interventions. The framework was developed by comparatively analyzing three large-scale NTD interventions in Eastern Africa: rabies elimination in Tanzania, sleeping sickness control in Uganda and the prevention of parasitic worms in Zambia. The framework includes five “intervention domains” where the effectiveness of these interventions was determined: 1) the terrain of intervention; 2) community agency; 3) the strategies and incentives of field staff; 4) the socio-materiality of technology; and 5) the governance of interventions. The paper illustrates the importance of each of these domains, presenting lessons learnt and practical recommendations. As a flexible analytical tool, the framework could be integrated into the planning and implementation process itself, bringing the insights of socio-anthropological approaches into an emerging science of NTD delivery.

Objectives Recently, improvements have been made to diagnostics for gambiense sleeping sickness control but their performance remains poorly documented and may depend on specimen processing prior to examination. In a prospective study in the Democratic Republic of the Congo, we compared the diagnostic performance of several parasite detection techniques, immune trypanolysis and of m18S PCR on whole blood stored in a stabilisation buffer or dried on filter paper. Methods Individuals with CATT whole blood (WB) titer ≥1∶4 or with clinical signs indicative for sleeping sickness were examined for presence of trypanosomes in lymph node aspirate (LNA) and/or in blood. Blood was examined with Capillary Centrifugation Technique (CTC), mini-Anion Exchange Centrifugation Technique (mAECT) and mAECT on buffy coat (BC). PCR was performed on whole blood (i) stored in guanidine hydrochloride EDTA (GE) stabilisation buffer and (ii) dried on filter paper, and repeatability and reproducibility were assessed. Immune trypanolysis (TL) was performed on plasma. Results A total of 237 persons were included. Among 143 parasitologically confirmed cases, 85.3% had a CATT-WB titre of ≥1/8, 39.2% were positive in LNA, 47.5% in CTC, 80.4% in mAECT-WB, 90.9% in mAECT-BC, 95.1% in TL and up to 89.5% in PCR on GE-stabilised blood. PCR on GE-stabilised blood showed highest repeatability (87.8%) and inter-laboratory reproducibility (86.9%). Of the 94 non-confirmed suspects, respectively 39.4% and 23.4% were TL or PCR positive. Suboptimal specificity of PCR and TL was also suggested by latent class analysis. Conclusion The combination of LNA examination with mAECT-BC offered excellent diagnostic sensitivity. For PCR, storage of blood in stabilisation buffer is to be preferred over filter paper. TL as well as PCR are useful for remote diagnosis but are not more sensitive than mAECT-BC. For TL and PCR, the specificity, and thus usefulness for management of non-confirmed suspects remain to be determined., Author Summary Human African trypanosomiasis or sleeping sickness still causes considerable suffering in sub-Sahara Africa. Diagnostics for this infectious disease constantly improve but their performance in terms of accuracy and reproducibility should be evaluated prior to implementation in control activities. We evaluated the diagnostic performance of several microscopic, serological and molecular diagnostic tests on a cohort of 237 sleeping sickness suspects in the Democratic Republic of the Congo. Since molecular diagnostics are rather sophisticated, we also assessed their repeatability and reproducibility. In the absence of a golden standard test, latent class analysis revealed that the suboptimal specificity of the serological and molecular tests is an issue. Our study shows the superior diagnostic sensitivity of the combination of lymph node aspirate examination and separation of trypanosomes from blood by mini Anion Exchange Centrifugation Techniques.

15 páginas, 2 figuras, 2 tablas, Background Scientific literature on cystic echinococcosis (CE) reporting data on risk factors is limited and to the best of our knowledge, no global evaluation of human CE risk factors has to date been performed. This systematic review (SR) summarizes available data on statistically relevant potential risk factors (PRFs) associated with human CE., Methodology/Principal Findings Database searches identified 1,367 papers, of which thirty-seven were eligible for inclusion. Of these, eight and twenty-nine were case-control and cross-sectional studies, respectively. Among the eligible papers, twenty-one were included in the meta-analyses. Pooled odds ratio (OR) were used as a measure of effect and separately analysed for the two study designs. PRFs derived from case-control studies that were significantly associated with higher odds of outcome were ªdog free to roamº (OR 5.23; 95% CI 2.45±11.14), ªfeeding dogs with visceraº (OR 4.69; 95% CI 3.02±7.29), ªslaughter at homeº (OR 4.67; 95% CI 2.02±10.78) or at ªslaughterhousesº (OR 2.7; 95% CI 1.15±6.3), ªdog ownershipº (OR 3.54; 95% CI 1.27±9.85), ªliving in rural areasº (OR 1.83; 95% CI 1.16±2.9) and ªlow incomeº (OR 1.68; 95% CI 1.02±2.76). Statistically significant PRFs from cross-sectional studies with higher odds of outcome were ªage >16 yearsº (OR 6.08; 95% CI 4.05±9.13), ªliving in rural areasº (OR 2.26; 95% CI 1.41±3.61), ªbeing femaleº (OR 1.38; 95% CI 1.06± 1.8) and ªdog ownershipº (OR 1.37; 95% CI 1.01±1.86). Conclusions/Significance Living in endemic rural areas, in which free roaming dogs have access to offal and being a dog-owner, seem to be among the most significant PRFs for acquiring this parasiticinfection. Results of data analysed here may contribute to our understanding of the PRFs for CE and may potentially be useful in planning community interventions aimed at controlling CE in endemic areas., Conclusions/Significance Living in endemic rural areas, in which free roaming dogs have access to offal and being a dog-owner, seem to be among the most significant PRFs for acquiring this parasiticinfection. Results of data analysed here may contribute to our understanding of the PRFs for CE and may potentially be useful in planning community interventions aimed at controlling CE in endemic areas., This research received funding from the European Community's Seventh Framework Programme under the grant agreement 602051 (Project HERACLES: Human cystic Echinococcosis ReseArch in CentraL and Eastern Societies; http://www.heracles-fp7.eu/).

Rabies is a fatal zoonosis that still causes nearly 70, 000 human deaths every year. In Europe, the oral rabies vaccination (ORV) of red foxes (Vulpes vulpes) was developed in the late 1970s and has demonstrated its effectiveness in the eradication of the disease in Western and some Central European countries. Following the accession of the three Baltic countries—Estonia, Latvia and Lithuania—to the European Union in 2004, subsequent financial support has allowed the implementation of regular ORV campaigns since 2005–2006. This paper reviews ten years of surveillance efforts and ORV campaigns in these countries resulting in the near eradication of the disease. The various factors that may have influenced the results of vaccination monitoring were assessed using generalized linear models (GLMs) on bait uptake and on herd immunity. As shown in previous studies, juveniles had lower bait uptake level than adults. For the first time, raccoon dogs (Nyctereutes procyonoides) were shown to have significantly lower bait uptake proportion compared with red foxes. This result suggests potentially altered ORV effectiveness in this invasive species compared to the red foxes. An extensive phylogenetic analysis demonstrated that the North-East European (NEE) rabies phylogroup is endemic in all three Baltic countries. Although successive oral vaccination campaigns have substantially reduced the number of detected rabies cases, sporadic detection of the C lineage (European part of Russian phylogroup) underlines the risk of reintroduction via westward spread from bordering countries. Vaccine induced cases were also reported for the first time in non-target species (Martes martes and Meles meles)., Author Summary This paper reviews ten years of rabies epidemiology in the three Baltic countries. Both surveillance efforts and oral rabies vaccination campaigns have resulted in the near eradication of the disease. Multivariate analysis assessed with generalized linear models (GLM) suggested lower oral vaccination effectiveness in raccoon dogs compared with red foxes, highlighting the importance of adapting oral vaccination strategy to each vector of the disease. Although eradication of the disease is almost achieved, the detection of some cases belonging with the Russian rabies lineage emphasizes a risk of rabies reintroduction in the Baltic States due to westward spread from bordering countries. This study show also the first vaccine-induced cases detected in non-target species (Martes martes and Meles meles).

Background Despite the availability of effective interventions and public recognition of the severity of the problem, rabies continues to suffer neglect by programme planners in India and other low and middle income countries. We investigate whether this state of ‘policy impasse’ is due to, at least in part, the research community not catering to the information needs of the policy makers. Methods & Findings Our objective was to review the research output on rabies from India and examine its alignment with national policy priorities. A systematic literature review of all rabies research articles published from India between 2001 and 2011 was conducted. The distribution of conducted research was compared to the findings of an earlier research prioritization exercise. It was found that a total of 93 research articles were published from India since 2001, out of which 61% consisted of laboratory based studies focussing on rabies virus. Animals were the least studied group, comprising only 8% of the research output. One third of the articles were published in three journals focussing on vaccines and infectious disease epidemiology and the top 4 institutions (2 each from the animal and human health sectors) collectively produced 49% of the national research output. Biomedical research related to development of new interventions dominated the total output as opposed to the identified priority domains of socio-politic-economic research, basic epidemiological research and research to improve existing interventions. Conclusion The paper highlights the gaps between rabies research and policy needs, and makes the case for developing a strategic research agenda that focusses on rabies control as an expected outcome., Author Summary Rabies is among the most widely spread zoonoses (diseases that are naturally transmitted between vertebrate animals and humans) in humans in most Asian, African and Latin American countries. Even though researchers have demonstrated effectiveness of strategies to control rabies at the population level, such as post exposure prophylaxis in humans and animal birth control and immunization among dogs, are well known, policy makers in most countries are hesitant to implement these strategies. This paper examines the disconnect that prevents the translation of scientific research outputs into effective policies. We contrasted the type of research papers published on rabies from India in the last eleven years with a previously identified set of priority research options. We found that most published research articles related to biomedical research focussing on development of new interventions. This was in contrast to policy and systems-related research and research to improve the performance of existing interventions that were identified as priority research options for India earlier. The findings of our study highlight the importance of moving beyond a purely researcher-driven agenda and suggest the need to promote research that has a vision of rabies control in the near future.

Background An outbreak of dengue fever (DF) occurred in Guangdong Province, China in 2013 with the highest number of cases observed within the preceding ten years. DF cases were clustered in the Pearl River Delta economic zone (PRD) in Guangdong Province, which accounted for 99.6% of all cases in Guangdong province in 2013. The main vector in PRD was Aedes albopictus. We investigated the socioeconomic and environmental factors at the township level and explored how the independent variables jointly affect the DF epidemic in the PRD. Methodology/Principal Findings Six factors associated with the incidence of DF were identified in this project, representing the urbanization, poverty, accessibility and vegetation, and were considered to be core contributors to the occurrence of DF from the perspective of the social economy and the environment. Analyses were performed with Generalized Additive Models (GAM) to fit parametric and non-parametric functions to the relationships between the response and predictors. We used a spline-smooth technique and plotted the predicted against the observed co-variable value. The distribution of DF cases was over-dispersed and fit the negative binomial function better. The effects of all six socioeconomic and environmental variables were found to be significant at the 0.001 level and the model explained 45.1% of the deviance by DF incidence. There was a higher risk of DF infection among people living at the prefectural boundary or in the urban areas than among those living in other areas in the PRD. The relative risk of living at the prefectural boundary was higher than that of living in the urban areas. The associations between the DF cases and population density, GDP per capita, road density, and NDVI were nonlinear. In general, higher “road density” or lower “GDP per capita” were considered to be consistent risk factors. Moreover, higher or lower values of “population density” and “NDVI” could result in an increase in DF cases. Conclusion In this study, we presented an effect analysis of socioeconomic and environmental factors on DF occurrence at the smallest administrative unit (township level) for the first time in China. GAM was used to effectively detect the nonlinear impact of the predictors on the outcome. The results showed that the relative importance of different risk factors may vary across the PRD. This work improves our understanding of the differences and effects of socioeconomic and environmental factors on DF and supports effectively targeted prevention and control measures., Author Summary Dengue fever is an infectious disease transmitted by mosquitoes. It is a major public health problem in tropical and subtropical regions around the world. Dengue fever is of great interest in the Pearl River Delta economic zone (PRD) of Guangdong province, China because the outbreak in 2013 was the largest in the previous 10 years. Due to the low degree of diversity in the climatic conditions in the PRD, socioeconomic and environmental factors may be the major contributing factors. The objective of this paper was to perform an assessment and detect the socioeconomic and environmental impact on cases at the smallest administrative unit (the township level). Six factors were identified in this work, representing urbanization, poverty, accessibility and vegetation. The effects of all these factors were found to be significant. The results showed that the relative importance of different risk factors may vary across the PRD. The higher risk areas and vulnerable populations identified in this paper will provide guidance for public health practitioners to create targeted, strategic plans and implement effective public health prevention and control measures.

Introduction In Colombia, organochloride, organophosphate, carbamate, and pyrethroid insecticides are broadly used to control Aedes aegypti populations. However, Colombian mosquito populations have shown variability in their susceptibility profiles to these insecticides, with some expressing high resistance levels. Materials and methods In this study, we analyzed the susceptibility status of ten Colombian field populations of Ae. aegypti to two pyrethroids; permethrin (type-I pyrethroid) and lambda-cyhalothrin (type-II pyrethroid). In addition, we evaluated if mosquitoes pressured with increasing lambda-cyhalothrin concentrations during some filial generations exhibited altered allelic frequency of these kdr mutations and the activity levels of some metabolic enzymes. Results Mosquitoes from all field populations showed resistance to lambda-cyhalothrin and permethrin. We found that resistance profiles could only be partially explained by kdr mutations and altered enzymatic activities such as esterases and mixed-function oxidases, indicating that other yet unknown mechanisms could be involved. The molecular and biochemical analyses of the most pyrethroid-resistant mosquito population (Acacías) indicated that kdr mutations and altered metabolic enzyme activity are involved in the resistance phenotype expression. Conclusions In this context, we propose genetic surveillance of the mosquito populations to monitor the emergence of resistance as an excellent initiative to improve mosquito-borne disease control measures., Author summary The main method of preventing Aedes-borne diseases such as dengue, Zika, and chikungunya is by targeting the primary mosquito vector, Aedes aegypti, with insecticides. However, the success of these vector control strategies is jeopardized by the widespread development of insecticide resistance in mosquito populations. Furthermore, the molecular mechanisms of insecticide resistance in Ae. aegypti are still not well understood, resulting in limited resistance mitigation and management strategies. In this paper, we found that resistance to some pyrethroid insecticides in different Colombian cities is associated with three allelic substitutions V419L, V1016I, and F1534C, on the voltage-gated sodium channel gene, known as kdr (‘knock-down resistance’) mutations, with all three mutations present in mosquitoes resistant to pyrethroids. The data also showed that kdr mutations are important in conferring low resistance levels, but after around 10-fold intensity, the allele frequencies don’t change, indicating that other mechanisms contribute to the resistance. Thus, we found that mosquitoes under selective pressure with insecticides present also altered enzymatic activities such as esterases and mixed-function oxidases, indicating that kdr mutations and metabolic enzymes are involved in the resistance expression. The findings on the extent of insecticide resistance and the molecular mechanisms underpinning the problem will impact the surveillance, selection, and rational use of insecticides by local health authorities.

Background The Wolbachia strategy aims to manipulate mosquito populations to make them incapable of transmitting dengue viruses between people. To test its efficacy, this strategy requires field trials. Public consultation and engagement are recognized as critical to the future success of these programs, but questions remain regarding how to proceed. This paper reports on a case study where social research was used to design a community engagement framework for a new dengue control method, at a potential release site in central Vietnam. Methodology/Principal Findings The approach described here, draws on an anthropological methodology and uses both qualitative and quantitative methods to design an engagement framework tailored to the concerns, expectations, and socio-political setting of a potential trial release site for Wolbachia-infected Aedes aegypti mosquitoes. The process, research activities, key findings and how these were responded to are described. Safety of the method to humans and the environment was the most common and significant concern, followed by efficacy and impact on local lives. Residents expected to be fully informed and engaged about the science, the project, its safety, the release and who would be responsible should something go wrong. They desired a level of engagement that included regular updates and authorization from government and at least one member of every household at the release site. Conclusions/Significance Results demonstrate that social research can provide important and reliable insights into public concerns and expectations at a potential release site, as well as guidance on how these might be addressed. Findings support the argument that using research to develop more targeted, engagement frameworks can lead to more sensitive, thorough, culturally comprehensible and therefore ethical consultation processes. This approach has now been used successfully to seek public input and eventually support for releases Wolbachia-infected mosquitoes, in two different international settings - Australia and Vietnam., Author Summary In recent years, a number of new strategies using novel technologies for the control of dengue fever control have emerged. These strategies are notably different from their predecessors and not without controversy. Many also require open release field trials to test their efficacy. Public consultation and engagement are recognized as critical to the future success of these programs, but questions remain regarding how to proceed. In this paper we describe an approach to public engagement that uses social research to design an engagement framework and communication materials tailored to the concerns, expectations, and socio-political setting of potential trial release sites. This approach was developed and implemented in Australia (2008–2010) where the first publicly supported field trials occurred January 2011. We report here on the implementation of this approach in Vietnam (2009–2010) where the second release will occur in 2014. This paper describes the process, research activities, outcomes and key findings from the Vietnamese field site. It highlights key public concerns and expectations about engagement and authorization and shows how these were used to develop a more targeted, culturally appropriate and comprehensible engagement framework and communication materials. The paper demonstrates the viability of this approach to community engagement for new dengue control strategies, in a ‘developing’ country context.

Background Despite doubts about methods used and the association between vector density and dengue transmission, routine sampling of mosquito vector populations is common in dengue-endemic countries worldwide. This study examined the evidence from published studies for the existence of any quantitative relationship between vector indices and dengue cases. Methodology/Principal Findings From a total of 1205 papers identified in database searches following Cochrane and PRISMA Group guidelines, 18 were included for review. Eligibility criteria included 3-month study duration and dengue case confirmation by WHO case definition and/or serology. A range of designs were seen, particularly in spatial sampling and analyses, and all but 3 were classed as weak study designs. Eleven of eighteen studies generated Stegomyia indices from combined larval and pupal data. Adult vector data were reported in only three studies. Of thirteen studies that investigated associations between vector indices and dengue cases, 4 reported positive correlations, 4 found no correlation and 5 reported ambiguous or inconclusive associations. Six out of 7 studies that measured Breteau Indices reported dengue transmission at levels below the currently accepted threshold of 5. Conclusions/Significance There was little evidence of quantifiable associations between vector indices and dengue transmission that could reliably be used for outbreak prediction. This review highlighted the need for standardized sampling protocols that adequately consider dengue spatial heterogeneity. Recommendations for more appropriately designed studies include: standardized study design to elucidate the relationship between vector abundance and dengue transmission; adult mosquito sampling should be routine; single values of Breteau or other indices are not reliable universal dengue transmission thresholds; better knowledge of vector ecology is required., Author Summary Routine sampling of mosquito vector populations is common in dengue-endemic countries worldwide despite doubts about methods used or the correlation between vector density and dengue transmission. This systematic review examined the published evidence investigating associations between vector indices and dengue cases. From a total of 1205 papers identified in database searches, 18 were included for review. A range of designs were seen, particularly in spatial sampling and analyses, and all but 3 were classed as weak study designs. Thirteen studies investigated associations between vector indices and dengue cases: 4 reported positive correlations, 4 found no correlation and 5 reported ambiguous/unreliable associations. Of 7 studies that measured the Breteau Index, 6 reported dengue transmission at levels below the currently accepted threshold of 5. There was little evidence of quantifiable associations between vector indices and dengue transmission that could reliably be used to predict outbreaks. Furthermore, appropriately designed studies are required to elucidate the relationship between vector abundance and dengue transmission. Recommendations include: standardizing study designs, particularly with respect to spatial heterogeneity; vector surveillance programs should sample adult mosquitoes; global values of the Breteau Index are not reliable universal dengue transmission thresholds; and better knowledge of vector ecology is required.

West Nile virus (WNV) is a globally distributed mosquito-borne virus of great public health concern. The number of WNV human cases and mosquito infection patterns vary in space and time. Many statistical models have been developed to understand and predict WNV geographic and temporal dynamics. However, these modeling efforts have been disjointed with little model comparison and inconsistent validation. In this paper, we describe a framework to unify and standardize WNV modeling efforts nationwide. WNV risk, detection, or warning models for this review were solicited from active research groups working in different regions of the United States. A total of 13 models were selected and described. The spatial and temporal scales of each model were compared to guide the timing and the locations for mosquito and virus surveillance, to support mosquito vector control decisions, and to assist in conducting public health outreach campaigns at multiple scales of decision-making. Our overarching goal is to bridge the existing gap between model development, which is usually conducted as an academic exercise, and practical model applications, which occur at state, tribal, local, or territorial public health and mosquito control agency levels. The proposed model assessment and comparison framework helps clarify the value of individual models for decision-making and identifies the appropriate temporal and spatial scope of each model. This qualitative evaluation clearly identifies gaps in linking models to applied decisions and sets the stage for a quantitative comparison of models. Specifically, whereas many coarse-grained models (county resolution or greater) have been developed, the greatest need is for fine-grained, short-term planning models (m–km, days–weeks) that remain scarce. We further recommend quantifying the value of information for each decision to identify decisions that would benefit most from model input.

Background Echinococcosis is a complex zoonosis that has domestic and sylvatic lifecycles, and a range of different intermediate and definitive host species. The complexities of its transmission and the sparse evidence on the effectiveness of control strategies in diverse settings provide significant challenges for the design of effective public health policy against this disease. Mathematical modelling is a useful tool for simulating control packages under locally specific transmission conditions to inform optimal timing and frequency of phased interventions for cost-effective control of echinococcosis. The aims of this review of 30 years of Echinococcus modelling were to discern the epidemiological mechanisms underpinning models of Echinococcus granulosus and E. multilocularis transmission and to establish the need to include a human transmission component in such models. Methodology/Principal Findings A search was conducted of all relevant articles published up until July 2012, identified from the PubMED, Web of Knowledge and Medline databases and review of bibliographies of selected papers. Papers eligible for inclusion were those describing the design of a new model, or modification of an existing mathematical model of E. granulosus or E. multilocularis transmission. A total of 13 eligible papers were identified, five of which described mathematical models of E. granulosus and eight that described E. multilocularis transmission. These models varied primarily on the basis of six key mechanisms that all have the capacity to modulate model dynamics, qualitatively affecting projections. These are: 1) the inclusion of a ‘latent’ class and/or time delay from host exposure to infectiousness; 2) an age structure for animal hosts; 3) the presence of density-dependent constraints; 4) accounting for seasonality; 5) stochastic parameters; and 6) inclusion of spatial and risk structures. Conclusions/Significance This review discusses the conditions under which these mechanisms may be important for inclusion in models of Echinococcus transmission and proposes recommendations for the design of dynamic human models of transmission. Accounting for the dynamic behaviour of the Echinococcus parasites in humans will be key to predicting changes in the disease burden over time and to simulate control strategies that optimise public health impact., Author Summary Echinococcosis is a complex zoonosis for which there is sparse evidence on the effectiveness of control strategies in diverse settings. This presents significant challenges for the design of effective public health policy against this disease. Mathematical modelling is a useful tool for simulating control packages under locally specific transmission conditions to inform optimal timing and frequency of phased interventions for cost-effective control of echinococcosis. This systematic review of 30 years of Echinococcus modelling discusses the importance of six key epidemiological mechanisms underpinning models of Echinococcus granulosus and E. multilocularis transmission and establishes the need to include a human transmission component. Accounting for the dynamic behaviour of the Echinococcus parasites in humans will be key to predicting changes in the disease burden over time and to simulate control strategies that optimise public health impact.

Background One Health is particularly relevant to the Horn of Africa where many people’s livelihoods are highly dependent on livestock and their shared environment. In this context, zoonoses may have a dramatic impact on both human and animal health, but also on country economies. This scoping review aimed to characterise and evaluate the nature of zoonotic disease research in the Horn region. Specifically, it addressed the following questions: (i) what specific zoonotic diseases have been prioritised for research, (ii) what data have been reported (human, animal or environment), (iii) what methods have been applied, and (iv) who has been doing the research? Methodology/principal findings We used keyword combinations to search online databases for peer-reviewed papers and theses. Screening and data extraction (disease, country, domain and method) was performed using DistillerSR. A total of 2055 studies focusing on seven countries and over 60 zoonoses were included. Brucellosis attracted the highest attention in terms of research while anthrax, Q fever and leptospirosis have been comparatively under-studied. Research efforts did not always align with zoonoses priorities identified at national levels. Despite zoonoses being a clear target for ‘One Health’ research, a very limited proportion of studies report data on the three domains of human, animal and environment. Descriptive and observational epidemiological studies were dominant and only a low proportion of publications were multidisciplinary. Finally, we found that a minority of international collaborations were between Global South countries with a high proportion of authors having affiliations from outside the Horn of Africa. Conclusions/significance There is a growing interest in zoonoses research in the Horn of Africa. Recommendations arising from this scoping review include: (i) ensuring zoonoses research aligns with national and global research agendas; (ii) encouraging researchers to adopt a holistic, transdisciplinary One Health approach following high quality reporting standards (COHERE, PRISMA, etc.); and (iii) empowering local researchers supported by regional and international partnerships to engage in zoonoses research., Author summary Zoonoses are diseases that are transmissible between animals and humans. Some emerging or re-emerging zoonoses, like avian influenza, regularly make the headlines in international media. Others, like rabies or echinococcosis, which mainly affect poor communities, attract much less attention, and are considered neglected by the World Health Organisation (WHO). In the Horn of Africa, many people live in close proximity to livestock and depend on them for food and income. Their frequent interaction with animals increases the risk of contracting zoonoses. In our work, we have searched for existing research publications on zoonoses in the Horn of Africa to guide future research on most neglected areas. Based on 2055 publications, we have described which zoonoses have been studied where and using which method. Notably, we found that very limited research followed One Health approaches. That implies that separate focus was given to animals or humans and a single method or discipline was used, while the One Health approach advocates for multidisciplinary and multisectoral collaboration to address complex issues like zoonoses. Finally, we identified that a majority of authors were affiliated with countries from the Global North which hinders relevance, equity and sustainability of Global North-Global South research collaborations.

Background In recent years, new strategies aimed at reducing the capacity of mosquito vectors to transmit dengue fever have emerged. As with earlier control methods, they will have to be employed in a diverse range of communities across the globe and into the main settings for disease transmission, the homes, businesses and public buildings of residents in dengue-affected areas. However, these strategies are notably different from previous methods and draw on technologies that are not without controversy. Public engagement and authorization are critical to the future success of these programs. Methodology/Principal Findings This paper reports on an Australian case study where long-term social research was used to enable participation and the design of an engagement strategy tailored specifically to the sociopolitical setting of a potential trial release site of Wolbachia-infected Aedes aegytpi mosquitoes. Central themes of the social research, methods used and conclusions drawn are briefly described. Results indicate that different communities are likely to have divergent expectations, concerns and cultural sensibilities with regard to participation, engagement and authorization. Conclusions/Significance The findings show that a range of issues need to be understood and taken into account to enable sensitive, ethical and effective engagement when seeking public support for new dengue control methods., Author Summary A number of new strategies are emerging to combat dengue fever. Many are notably different from earlier methods and not without controversy. Public participation, engagement, enablement and authorization are recognized as critical to the success of field trials and future use and acceptability of new strategies. It is well established in cultural anthropology and public health that interventions often fail because of a lack of understanding of the local context, lay knowledge of the disease, and community, local or regional expectations around engagement. In this paper we demonstrate the critical importance of using long-term social research to address these issues and to design engagement strategies and communication materials that are tailored specifically to the needs, expectations and concerns of residents at a potential release site. We report on the results of an extensive and systematic social research and engagement program undertaken in the two years prior to the first successful release of Wolbachia-infected Aedes aegypti mosquitoes in Cairns, northern Australia.

BackgroundWolbachia is an endosymbiont bacterium generally found in about 40% of insects, including mosquitoes, but it is absent in Aedes aegypti which is an important vector of several arboviral diseases. The evidence that Wolbachia trans-infected Ae. aegypti mosquitoes lost their vectorial competence and became less capable of transmitting arboviruses to human hosts highlights the potential of using Wolbachia- based approaches for prevention and control of arboviral diseases. Recently, release of Wolbachia trans-infected Ae. aegypti has been deployed widely in many countries for the control of mosquito-borne viral diseases. Field surveillance and monitoring of Wolbachia presence in released mosquitoes is important for the success of these control programs. So far, a number of studies have reported the development of loop mediated isothermal amplification (LAMP) assays to detect Wolbachia in mosquitoes, but the methods still have some specificity issues.Methodology/Principal FindingsWe describe here the development of a LAMP combined with the DNA strand displacement-based electrochemical sensor (BIOSENSOR) method to detect wAlbB Wolbachia in trans-infected Ae. aegypti. Our developed LAMP primers were more specific to wAlbB detection than those of the previous published ones if the assays were conducted with low-cost and non-specific detecting dyes. The detection capacity of our LAMP technique was 3.8 nM and the detection limit reduced to 2.16 fM when combined with the BIOSENSOR. Our study demonstrates that the BIOSENSOR can also be applied as a stand-alone method for detecting Wolbachia; and it showed high sensitivity when used with the crude DNA extracts of macerated mosquito samples without DNA purification.Conclusions/SignificanceOur results suggest that both LAMP and BIOSENSOR, either used in combination or stand-alone, are robust and sensitive. The methods have good potential for routine detection of Wolbachia in mosquitoes during field surveillance and monitoring of Wolbachia-based release programs, especially in countries with limited resources.Author SummaryMosquito-borne diseases such as dengue, chikungunya, Zika, and yellow fever are transmitted to humans mainly by the bites of Aedes aegypti mosquitoes. Controlling these diseases relies mostly on the use of insecticides, in which the efficiency has been reduced through development of insecticide resistance in mosquitoes. Wolbachia is the endosymbiotic bacteria that are naturally found in 40% of insects, including mosquitoes. The bacteria could protect their hosts from viral infections and could also cause sterility in host populations, therefore, providing an opportunity to use them for disease control. Application of a Wolbachia-based strategy needs simple, rapid and sensitive methods for detecting the bacteria in released mosquitoes. In this paper, we develop the combined methods of LAMP and BIOSENSORS for detecting wAlbB Wolbachia in mosquitoes. Our positive LAMP reaction can be visualized by color change from violet to blue at a sensitivity of ≥ 60 pg of genomic DNA. When used in combination with the BIOSENSOR method, the sensitivity increased a million fold without losing specificity. Our study indicates that both developed methods, either used in combination or stand-alone, are efficient and cost-effective, hence, it could be applied for routine surveys of Wolbachia in mosquito control programs that use Wolbachia-based approaches.

Public health institutions with sectorized structure and low integration among field teams, old-fashioned practices such as paper-based storage system, and poorly qualified health agents have limited ability to conduct accurate surveillance and design effective timely interventions. Herein, we describe the steps taken by the Zoonosis Control Center of Foz do Iguaçu (CCZ-Foz) in the last 23 years to move from an archaic and sectorized structure to a modern and timely surveillance program embracing zoonotic diseases, venomous animal injuries, and vector-borne diseases epidemiology under the One Health approach. The full implementation of the One Health approach was based on 5 axes: (1) merging sectorized field teams; (2) adoption of digital solutions; (3) health agents empowerment and permanent capacitation; (4) social mobilization; and (5) active surveys. By doing so, notifications related to zoonotic diseases and venomous animals increased 10 and 21 times, respectively, with no impairment on arbovirus surveillance (major concern in the city). Open sources database (PostgreSQL) and software (QGis) are daily updated and create real-time maps to support timely decisions. The adoption of One Health approach increased preparedness for endemic diseases and reemerging and emerging threats such as Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2).

Background The development of insecticide resistance in mosquitoes can have pleiotropic effects on key behaviours such as mating competition and host-location. Documenting these effects is crucial for understanding the dynamics and costs of insecticide resistance and may give researchers an evidence base for promoting vector control programs that aim to restore or conserve insecticide susceptibility. Methods and findings We evaluated changes in behaviour in a backcrossed strain of Aedes aegypti, homozygous for two knockdown resistance (kdr) mutations (V1016G and S989P) isolated in an otherwise fully susceptible genetic background. We compared biting activity, host location behaviours, wing beat frequency (WBF) and mating competition between the backcrossed strain, and the fully susceptible and resistant parental strains from which it was derived. The presence of the homozygous kdr mutations did not have significant effects on blood avidity, the time to locate a host, or WBF in females. There was, however, a significant reduction in mean WBF in males and a significant reduction in estimated male mating success (17.3%), associated with the isolated kdr genotype. Conclusions Our results demonstrate a cost of insecticide resistance associated with an isolated kdr genotype and manifest as a reduction in male mating success. While there was no recorded difference in WBF between the females of our strains, the significant reduction in male WBF recorded in our backcrossed strain might contribute to mate-recognition and mating disruption. These consequences of resistance evolution, especially when combined with other pleiotropic fitness costs that have been previously described, may encourage reversion to susceptibility in the absence of insecticide selection pressures. This offers justification for the implementation of insecticide resistance management strategies based on the rotation or alternation of different insecticide classes in space and time., Author summary The mosquito Aedes aegypti is the main vector of dengue, chikungunya, and Zika. Its control relies heavily on the use of insecticides but the rapid evolution of resistance to these chemicals compromises their efficacy. The conservation or restoration of insecticide susceptibility in Ae. aegypti populations is therefore of great importance. Insecticide susceptibility can be encouraged if the evolution of resistance is accompanied by fitness costs that favour susceptible mosquitoes in the absence of insecticides. This paper documents the first report of a reduction in mating success directly associated with an isolated mutation that confers insecticide resistance in Ae. aegypti. This change in behaviour appears related to alterations in male wing-beat frequency. Our results provide evidence of behavioural changes related to insecticide resistance in Ae. aegypti, suggesting a competitive advantage of susceptible individuals in the absence of insecticides in the field.

The phrase “the 800-pound gorilla in the room” refers to an obvious problem that everyone knows exists but pretends or chooses to ignore. In my December 2007 PLoS Neglected Tropical Diseases editorial I wrote about an unseemly underbelly of poverty that exists in my country, the United States of America, and the unacceptable disease burden among our poor that results from such neglected diseases as toxocariasis, cysticercosis, and toxoplasmosis [1]. According to the Pan American Health Organization (PAHO) (the regional office of the World Health Organization in the Americas) the Latin American and Caribbean region suffers from much larger pockets of poverty, and with it endemic neglected tropical diseases (NTDs) [2]. While researching a review of the NTDs in Latin America, I was particularly struck by the disproportionate concentration of these conditions among the poor living in Brazil. Although there are no gorillas in the rooms of Brazil I have concluded that the NTDs represent an ominous giant anteater (Myrmecophaga tridactyla, the largest species of anteater found in Brazil and elsewhere in the American tropics) that requires notice, attention, and urgent action. According to the World Bank, 22% of the population of Latin America and the Caribbean lives on less than US$2 per day [3]. An identical percentage of Brazil's population is also impoverished [4]. However, based on the Gini coefficient index, a metric that measures inequality of income or wealth distribution, Brazil at 0.59 has one of the greatest disparities between wealthy and poor people anywhere in the world [5]. One of the ways that this inequality manifests is in a shockingly high burden of NTDs. The roughly 40 million Brasileiros who live on less than US$2 per day [4] account for one-third of all of the poor people living in the Latin American and Caribbean region [3], but they suffer disproportionately from NTDs. Shown in Table 1 is a summary of Brazil's NTD burden. Based on information available from published peer-reviewed papers in the biomedical literature, as well as publicly available Web sites from PAHO and WHO [6]–[13], most of the NTD disease burden in Latin America and the Caribbean now occurs in Brazil, including virtually all of the cases of blinding trachoma and leprosy, and the majority of ascariasis, dengue, hookworm infection, schistosomiasis, and visceral leishmaniasis. Practically speaking these data mean that most of Brazil's poorest 40 million people are infected with one or more NTD, especially either hookworms (Necator americanus) or roundworms (Ascaris lumbricoides) or both. For instance, in collaboration with the Centro des Pesquisas Rene Rachou of FIOCRUZ (Fundacao Oswaldo Cruz) we recently showed that 68% of some rural communities in Minas Gerais State are infected with hookworms [14], which cause anemia and malnutrition [15]. In these same poor people the presence of hookworms in their intestines promotes susceptibility to coinfections with other worms, especially Ascaris roundworms and schistosomes (14). Table 1 Burden of Neglected Tropical Diseases in Brazil Throughout Minas Gerais and neighboring Bahia states, hookworm is known as amarelao, referring to the yellow–sallow complexion that results from chronic infection. Hookworm and other important NTDs are also major public health problems among Brazil's indigenous people [16]–[19] and citizens of African descent [20]. Recently, an economist at the University of Chicago found that chronic hookworm infection during childhood reduces future wage-earning by 43% [21] meaning that hookworm represents an important reason why poor people become trapped in poverty. This is the very same hookworm that sapped the energy from Jeca Tatu, the laborer popularized by the Brazilian writer Monteiro Lobato in the early twentieth century. An important feature of almost all of the NTDs is that they not only occur in the poverty setting but also promote poverty [22]. In 2003, President Luiz Inacio Lula a Silva launched Zero Hunger, an ambitious $500 million antipoverty drive focused on malnutrition [23]. However, as the World Food Programme has already discovered [24], in the absence of concurrent NTD control, measures that feed the children often simply result in feeding the worms first. Concurrent NTD control would make great sense, as it is now ranked among the most cost-effective health measures [22], and at a 15%–30% rate of return, a highly cost-effective antipoverty measure as well [25],[26]. NTD control would represent one of the most efficient and effective mechanisms for lifting the 40 million most poor Brazilians out of poverty. Everything is in place for Brazil to launch a nationwide effort geared at solving one of its greatest health disparities. The country has a charismatic president who is committed to the poor; it has in FIOCRUZ and other health ministry agencies and its universities some of the best disease control experts anywhere in the world (many of whom serve on the Editorial Board of PLoS Neglected Tropical Diseases); and, through these same organizations and a sophisticated biotechnology manufacturing infrastructure at Instituto Butantan (Sao Paulo) and Bio-manguinhos (Rio de Janeiro), the ability to innovate and produce a new and improved generation of “antipoverty” vaccines for NTDs. These vaccines target dengue, hookworm, leishmaniasis, leptospirosis, schistosomiasis, and yellow fever [27],[28]. Together with new foundations committed to solving health and other disparities in the Americas and some of the highest net-worth individuals anywhere, Brazil is in an excellent position to establish vital public–private partnerships to control, or in some cases, eliminate its most important NTDs. It has already taken leadership in the elimination of its Chagas disease problem, formerly one of the most devastating NTDs in the Southern Cone of South America [29], and has made great strides in the control and elimination of lymphatic filariasis and onchocerciasis [2],[12]. Now, just as Jeca Tatu who, once cured of his hookworm, championed social change, so too a new Brazilian public–private partnership could one day eliminate its NTDs as a substantive means for reducing poverty and achieving important Millennium Development Goals (an international set of goals and targets for sustainable poverty reduction in developing countries by the year 2015). As we enter the season of Carnaval this is a special time to remember Brazil's poorest 40 million people. In the meantime, PLoS Neglected Tropical Diseases remains committed to receiving and reviewing papers on the NTDs of Brazil and helping to facilitate communication among the Brazilian scientific community.

Background Early warning systems (EWS) are management tools to predict the occurrence of epidemics of infectious diseases. While climate-based EWS have been developed for malaria, no standard protocol to evaluate and compare EWS has been proposed. Additionally, there are several neglected tropical diseases whose transmission is sensitive to environmental conditions, for which no EWS have been proposed, though they represent a large burden for the affected populations. Methodology/Principal Findings In the present paper, an overview of the available linear and non-linear tools to predict seasonal time series of diseases is presented. Also, a general methodology to compare and evaluate models for prediction is presented and illustrated using American cutaneous leishmaniasis, a neglected tropical disease, as an example. The comparison of the different models using the predictive R 2 for forecasts of “out-of-fit” data (data that has not been used to fit the models) shows that for the several linear and non-linear models tested, the best results were obtained for seasonal autoregressive (SAR) models that incorporate climatic covariates. An additional bootstrapping experiment shows that the relationship of the disease time series with the climatic covariates is strong and consistent for the SAR modeling approach. While the autoregressive part of the model is not significant, the exogenous forcing due to climate is always statistically significant. Prediction accuracy can vary from 50% to over 80% for disease burden at time scales of one year or shorter. Conclusions/Significance This study illustrates a protocol for the development of EWS that includes three main steps: (i) the fitting of different models using several methodologies, (ii) the comparison of models based on the predictability of “out-of-fit” data, and (iii) the assessment of the robustness of the relationship between the disease and the variables in the model selected as best with an objective criterion., Author Summary Early Warning Systems (EWS) are management tools to predict the occurrence of epidemics. They are based on the dependence of a given infectious disease on environmental variables. Although several neglected tropical diseases are sensitive to the effect of climate, our ability to predict their dynamics has been barely studied. In this paper, we use several models to determine if the relationship between cases and climatic variability is robust—that is, not simply an artifact of model choice. We propose that EWS should be based on results from several models that are to be compared in terms of their ability to predict future number of cases. We use a specific metric for this comparison known as the predictive R 2, which measures the accuracy of the predictions. For example, an R 2 of 1 indicates perfect accuracy for predictions that perfectly match observed cases. For cutaneous leishmaniasis, R 2 values range from 72% to77%, well above predictions using mean seasonal values (64%). We emphasize that predictability should be evaluated with observations that have not been used to fit the model. Finally, we argue that EWS should incorporate climatic variables that are known to have a consistent relationship with the number of observed cases.

Background Like many countries from the Americas, Cuba is threatened by Aedes aegypti-associated arboviruses such as dengue (DENV), Zika (ZIKV), and chikungunya (CHIKV) viruses. Curiously, when CHIKV was actively circulating in the region in 2013–2014, no autochthonous transmission of this virus was detected in Havana, Cuba, despite the importation of chikungunya cases into this city. To investigate if the transmission ability of local mosquito populations could explain this epidemiological scenario, we evaluated for the first time the vector competence of two Ae. aegypti populations (Pasteur and Párraga) collected from Havana for dengue virus type 1 (DENV-1), CHIKV, and ZIKV. Methodology/Principal findings Mosquito populations were fed separately using blood containing ZIKV, DENV-1, or CHIKV. Infection, dissemination, and transmission rates, were estimated at 3 (exclusively for CHIKV), 7, and 14 days post exposure (dpe) for each Ae. aegypti population-virus combination. Both mosquito populations were susceptible to DENV-1 and ZIKV, with viral infection and dissemination rates ranging from 24–97% and 6–67% respectively. In addition, CHIKV disseminated in both populations and was subsequently transmitted. Transmission rates were low (, Author summary Dengue (DENV), Zika (ZIKV), and chikungunya (CHIKV) viruses are mainly transmitted by Aedes aegypti mosquitoes and together caused near 10 million human cases in the last decade in the Americas. The diseases caused by these viruses share symptoms ranging from mild clinical manifestations to severe complications, including fatal outcomes. Cuba has not been exempted from these viruses and is frequently subjected to dengue and more recently by diseases caused by ZIKV infection. Surprisingly, despite multiple CHIKV outbreaks recorded in neighboring countries, local circulation has not been reported in Havana. Although Ae. aegypti has been implicated as the major DENV and ZIKV vector because of its wide distribution on the island, neither field nor experimental approaches have been used to demonstrate the ability of Cuban Ae. aegypti populations to transmit DENVs, ZIKV, and CHIKV. In this laboratory study, we demonstrated that two Ae. aegypti populations from Havana could become infected with and transmit these viruses, albeit at low rates. Our paper suggests that the absence of CHIKV circulation in Havana was not related to refractoriness of Ae. aegypti populations to this virus; therefore, vector control remains being essential in preventing future arboviruses outbreaks.

The Global Programme to Eliminate Lymphatic Filariasis has made considerable progress but is experiencing challenges in meeting targets in some countries. Recent World Health Organization guidelines have recommended two rounds of triple-drug therapy with ivermectin, diethylcarbamazine (DEC), and albendazole (IDA), in areas where mass drug administration (MDA) results with two drugs (DEC and albendazole) have been suboptimal, as is the case in Samoa. In August 2018, Samoa was the first country in the world to implement countrywide triple-drug MDA. This paper aims to describe Samoa’s experience with program coverage and adverse events (AEs) in the first round of triple-drug MDA. We conducted a large cross-sectional community survey to assess MDA awareness, reach, compliance, coverage and AEs in September/October 2018, 7–11 weeks after the first round of triple-drug MDA. In our sample of 4420 people aged ≥2 years (2.2% of the population), age-adjusted estimates indicated that 89.0% of the eligible population were offered MDA, 83.9% of the eligible population took MDA (program coverage), and 80.2% of the total population took MDA (epidemiological coverage). Overall, 83.8% (2986/3563) reported that they did not feel unwell at all after taking MDA. Mild AEs (feeling unwell but able to do normal everyday things) were reported by 13.3% (476/3563) and moderate or severe AEs (feeling unwell and being unable to do normal everyday activities such as going to work or school) by 2.9% (103/3563) of participants. This study following the 2018 triple-drug MDA in Samoa demonstrated a high reported program awareness and reach of 90.8% and 89.0%, respectively. Age-adjusted program coverage of 83.9% of the total population showed that MDA was well accepted and well tolerated by the community., Author summary Lymphatic filariasis is a disease caused by worms and transmitted between humans by mosquitoes. A global program underway aims to eliminate lymphatic filariasis as a public health problem by distributing two deworming drugs to the whole population once a year for at least five years. In some countries, including Samoa, this strategy has not been sufficient to eliminate transmission. A new drug has been added, and in 2018, Samoa was the first country in the world to distribute country-wide triple-drug mass drug administration (MDA) using ivermectin, diethylcarbamazine, and albendazole. This study reports on the coverage achieved (percentage of people who reported taking the drugs) and adverse events after taking the drugs. The study was a large community survey of over 4000 people, done 7–11 weeks after the distribution of the first round of triple-drug MDA. We found that the program reached and offered MDA to approximately 90% of the whole population, and approximately 80% of the whole population swallowed the drugs. We also collected data on whether people felt unwell after taking the pills, and found that 84% of people taking the pills did not feel unwell at all, while 13% reported feeling unwell afterwards but were able to do normal everyday activities, and 3% reported that they had felt unwell and that it stopped them doing normal everyday activities, such as going to school or work.

Background Dengue (DENV), Ross River (RRV) and Barmah Forest viruses (BFV) are the most common human arboviral infections in Australia and the Pacific Island Countries and Territories (PICTs) and are associated with debilitating symptoms. All are nationally notifiable in Australia, but routine surveillance is limited to a few locations in the PICTs. Understanding the level of human exposure to these viruses can inform disease management and mitigation strategies. To assess the historic and current seroprevalence of DENV, RRV and BFV in Australia and the PICTs we conducted a systematic literature review of all published quantitative serosurveys. Methodology and principal findings The Preferred Reporting of Items for Systematic Reviews and Meta-Analyses procedures were adopted to produce a protocol to systematically search for published studies reporting the seroprevalence of DENV, RRV and BFV in Australia and the PICTs. Data for author, research year, location, study population, serosurvey methods and positive tests were extracted. A total of 41 papers, reporting 78 serosurveys of DENV, RRV and BFV including 62,327 samples met the inclusion criteria for this review. Seroprevalence varied depending on the assay used, strategy of sample collection and location of the study population. Significant differences were observed in reported seropositivity depending on the sample collection strategy with clinically targeted sampling reporting the highest seroprevalence across all three viruses. Non-stratified seroprevalence showed wide ranges in reported positivity with DENV 0.0% – 95.6%, RRV 0.0% – 100.0%, and BFV 0.3% – 12.5%. We discuss some of the causes of variation including serological methods used, selection bias in sample collection including clinical or environmental associations, and location of study site. We consider the extent to which serosurveys reflect the epidemiology of the viruses and provide broad recommendations regarding the conduct and reporting of arbovirus serosurveys. Conclusions and significance Human serosurveys provide important information on the extent of human exposure to arboviruses across: (1) time, (2) place, and (3) person (e.g., age, gender, clinical presentation etc). Interpreting results obtained at these scales has the potential to inform us about transmission cycles, improve diagnostic surveillance, and mitigate future outbreaks. Future research should streamline methods and reduce bias to allow a better understanding of the burden of these diseases and the factors associated with seroprevalence. Greater consideration should be given to the interpretation of seroprevalence in studies, and increased rigour applied in linking seroprevalence to transmission dynamics.

Background: American Samoa completed seven rounds of mass drug administration from 2000-2006 as part of the Global Programme to Eliminate Lymphatic Filariasis (LF). However, resurgence was confirmed in 2016 through a community-based survey and a WHO-recommended school-based transmission assessment survey (TAS). This paper uses data from the 2016 community survey to compare different spatial and non-spatial methods to characterise clustering and hotspots of LF.Method: Non-spatial clustering of infection markers (antigen [Ag], microfilaraemia [Mf], and antibodies (Ab [Wb123, Bm14, Bm33]) was assessed using intra-cluster correlation coefficients (ICC) at household and village levels. Spatial dependence, clustering and hotspots were explored using semivariograms, Kulldorf’s scan statistic and Getis-Ord Gi* statistics at locations of surveyed households.Results: The survey included 2671 persons (750 households, 730 unique locations in 30 villages). ICCs were higher at household (0.20-0.69) than village levels (0.10-0.30) for all infection markers. Semivariograms identified significant spatial dependency for all markers (range 207-562 metres). Using Kuldorff’s scan statistic, significant clustering was observed in two previously known locations of ongoing transmission: for all markers in Fagali’i and all Abs in Vaitogi. Getis-Ord Gi* statistic identified hotspots of all markers in Fagali’i, Vaitogi, and Pago Pago-Anua areas. A hotspot of Ag and Wb123 Ab was identified around the villages of Nua-Seetaga-Afao. Bm14 and Bm33 Ab hotspots were seen in Maleimi and Vaitogi-Ili’ili-Tafuna.Conclusion:Our study demonstrated the utility of different non-spatial and spatial methods for investigating clustering and hotspots, the benefits of using multiple infection markers, and the value of triangulating results between methods.Funding Statement: This work received financial support from the Coalition for Operational Research on Neglected Tropical Diseases (COR-NTD), which is funded at The Task Force for Global Health primarily by the Bill & Melinda Gates Foundation, by UK aid from the British government, and by the United States Agency for International Development through its Neglected Tropical Diseases Program. CLL was funded by an Australian National Health and Medical Research Council Fellowship (1109035). The funders had no role in interpretation of results, decision to publish, or preparation of the manuscript. MS is funded by a fellowship from the Westpac Scholars Trust.Declaration of Interests: The authors declare that they have no competing interests.Ethics Approval Statement: This study was approved by the American Samoa Institutional Review Board and the Human Research Ethics Committee at the Australian National University (protocol number 2016/482).

Background Like many infectious diseases, there is no practical gold standard for diagnosing clinical visceral leishmaniasis (VL). Latent class modeling has been proposed to estimate a latent gold standard for identifying disease. These proposed models for VL have leveraged information from diagnostic tests with dichotomous serological and PCR assays, but have not employed continuous diagnostic test information. Methods/Principal findings In this paper, we employ Bayesian latent class models to improve the identification of canine visceral leishmaniasis using the dichotomous PCR assay and the Dual Path Platform (DPP) serology test. The DPP test has historically been used as a dichotomous assay, but can also yield numerical information via the DPP reader. Using data collected from a cohort of hunting dogs across the United States, which were identified as having either negative or symptomatic disease, we evaluate the impact of including numerical DPP reader information as a proxy for immune response. We find that inclusion of DPP reader information allows us to illustrate changes in immune response as a function of age. Conclusions/Significance Utilization of continuous DPP reader information can improve the correct discrimination between individuals that are negative for disease and those with clinical VL. These models provide a promising avenue for diagnostic testing in contexts with multiple, imperfect diagnostic tests. Specifically, they can easily be applied to human visceral leishmaniasis when diagnostic test results are available. Also, appropriate diagnosis of canine visceral leishmaniasis has important consequences for curtailing spread of disease to humans.

Background As China is moving onto schistosomiasis elimination/eradication, diagnostic methods with both high sensitivity and specificity for Schistosoma japonicum infections in humans are urgently needed. Microscopic identification of eggs in stool is proven to have poor sensitivity in low endemic regions, and antibody tests are unable to distinguish between current and previous infections. Polymerase chain reaction (PCR) technologies for the detection of parasite DNA have been theoretically assumed to show high diagnostic sensitivity and specificity. However, the reported performance of PCR for detecting S. japonicum infection varied greatly among studies. Therefore, we performed a meta-analysis to evaluate the overall diagnostic performance of variable-temperature PCR technologies, based on stool or blood, for detecting S. japonicum infections in humans from endemic areas. Methods We searched literatures in eight electronic databases, published up to 20 January 2021. The heterogeneity and publication bias of included studies were assessed statistically. The risk of bias and applicability of each eligible study were assessed using the Quality Assessment of Diagnostic Accuracy Studies 2 tool (QUADAS-2). The bivariate mixed-effects model was applied to obtain the summary estimates of diagnostic performance. The hierarchical summary receiver operating characteristic (HSROC) curve was applied to visually display the results. Subgroup analyses and multivariate regression were performed to explore the source of heterogeneity. This research was performed following the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines and was registered prospectively in PROSPERO (CRD42021233165). Results A total of 2791 papers were retrieved. After assessing for duplications and eligilibity a total of thirteen publications were retained for inclusion. These included eligible data from 4268 participants across sixteen studies. High heterogeneity existed among studies, but no publication bias was found. The pooled analyses of PCR data from all included studies resulted in a sensitivity of 0.91 (95% CI: 0.83 to 0.96), specificity of 0.85 (95% CI: 0.65 to 0.94), positive likelihood ratio of 5.90 (95% CI: 2.40 to 14.60), negative likelihood ratio of 0.10 (95% CI: 0.05 to 0.20) and a diagnostics odds ratio of 58 (95% CI: 19 to 179). Case-control studies showed significantly better performances for PCR diagnostics than cross-sectional studies. This was further evidenced by multivariate analyses. The four types of PCR approaches identified (conventional PCR, qPCR, Droplet digital PCR and nested PCR) differed significantly, with nested PCRs showing the best performance. Conclusions Variable-temperature PCR has a satisfactory performance for diagnosing S. japonicum infections in humans in endemic areas. More high quality studies on S. japonicum diagnostic techniques, especially in low endemic areas and for the detection of dual-sex and single-sex infections are required. These will likely need to optimise a nested PCR alongside a highly sensitive gene target. They will contribute to successfully monitoring endemic areas as they move towards the WHO 2030 targets, as well as ultimately helping areas to achieve these goals., Author summary Schistosoma japonicum is a parasite that can cause serious intestinal schistosomiasis. The infection is mainly diagnosed by parasitological and immunological methods, such as Kato-Katz test and indirect hemagglutination assay. However, both of these are not sensitive enough to accurately assess schistosomiasis elimination/eradication. PCR assays, detecting parasite DNA, are theoretically a highly suitable, sensitive and specific, alternative. However, reported performance varies greatly among studies. Therefore, we performed this meta-analysis (PROSPERO, registered No. CRD42021233165) to analyze and summarize the data from relevant studies of variable-temperature PCR for the diagnosis of S. japonicum infections, using stool or blood samples from humans in endemic areas. We retrieved a total of 13 eligible articles including data from 4268 participants across 16 studies. There was high heterogeneity among studies, but no publication bias was found. Analyses revealed that PCR techniques had a satisfactory performance for diagnosing S. japonicum infections in humans in endemic areas, with both high sensitivity and specificity. Further research on S. japonicum diagnostic techniques, especially in low endemicity areas and for detection of dual-sex and single-sex infections are required. These may be best using highly sensitive gene targets in nested PCR reactions.

Cutaneous leishmaniasis (CL) due to Leishmania tropica is a neglected tropical disease characterized by a wide geographical distribution in the Mediterranean basin and is endemic in several of its countries. In addition, the vector Phlebotomus sergenti is abundantly present all around the basin. Its transmission cycle is still subject to debate. In some countries, the presence of an animal reservoir has been confirmed. In Morocco, CL due to L. tropica has risen since the 1980s and has spread widely to become the most abundant form of leishmaniasis in the territory. However, the anthroponotic transmission is so far the only recognized mode, despite recordings of L. tropica infection in animal hosts. In this review article, we assess the situation of CL due to L. tropica in the Mediterranean basin with a focus on Morocco and gather knowledge about any potential zoonotic transmission in the country. A concomitant zoonotic transmission could explain the persistence of the disease in areas where human protective measures combined with vector management did not help reduce the disease burden., Author summary Cutaneous leishmaniasis (CL) due to Leishmania tropica is a neglected tropical disease still considered to be anthroponotic in Morocco despite evidence of zoonotic transmission in other endemic countries of the Mediterranean basin. Its high incidence in the country makes it necessary to investigate whether or not a potential concomitant zoonotic transmission exists alongside the anthroponotic one. Moreover, several reports of natural animal infection have been noted in the country. In this paper, we review how CL due to L. tropica is present around the Mediterranean basin, with a particular focus on the Moroccan foci, and try to unveil the existence of a zoonotic transmission in the area. A deep understanding of the transmission patterns of L. tropica in the region is necessary for the implementation of effective control measures and a better assessment of the disease burden. In addition, the complexity of this neglected disease requires more fundamental and applied research to unveil potential factors involved in its spread.

Background Anthrax is a zoonotic disease that can be transmitted to humans from infected animals. During May–June 2017, three persons with probable cutaneous anthrax were reported in Arua District, Uganda; one died. All had recently handled carcasses of livestock that died suddenly and a skin lesion from a deceased person tested positive by PCR for Bacillus anthracis. During July, a bull in the same community died suddenly and the blood sample tested positive by PCR for Bacillus anthracis. The aim of this investigation was to establish the scope of the problem, identify exposures associated with illness, and recommend evidence-based control measures. Methods A probable case was defined as acute onset of a papulo-vesicular skin lesion subsequently forming an eschar in a resident of Arua District during January 2015–August 2017. A confirmed case was a probable case with a skin sample testing positive by polymerase chain reaction (PCR) for B. anthracis. Cases were identified by medical record review and active community search. In a case-control study, exposures between case-patients and frequency- and village-matched asymptomatic controls were compared. Key animal health staff were interviewed to learn about livestock deaths. Results There were 68 case-patients (67 probable, 1 confirmed), and 2 deaths identified. Cases occurred throughout the three-year period, peaking during dry seasons. All cases occurred following sudden livestock deaths in the villages. Case-patients came from two neighboring sub-counties: Rigbo (attack rate (AR) = 21.9/10,000 population) and Rhino Camp (AR = 1.9/10,000). Males (AR = 24.9/10,000) were more affected than females (AR = 0.7/10,000). Persons aged 30–39 years (AR = 40.1/10,000 population) were most affected. Among all cases and 136 controls, skinning (ORM-H = 5.0, 95%CI: 2.3–11), butchering (ORM-H = 22, 95%CI: 5.5–89), and carrying the carcass of livestock that died suddenly (ORM-H = 6.9, 95%CI: 3.0–16) were associated with illness. Conclusions Exposure to carcasses of animals that died suddenly was a likely risk factor for cutaneous anthrax in Arua District during 2015–2017. The recommendations are investigation of anthrax burden in livestock, prevention of animal infections through vaccinations, safe disposal of the carcasses, public education on risk factors for infection and prompt treatment of illness following exposure to animals that died suddenly., Author summary We are honored to submit an original research article entitled “Cutaneous anthrax associated with handling carcasses of animals that died suddenly of unknown cause: Arua District, Uganda, January 2015–August 2017”. Anthrax is a vaccine-preventable disease that can be transmitted to humans from infected animals. Our paper is based on 68 cutaneous anthrax case-patients and cases occurred throughout the three-year period, peaking during dry seasons in Arua District of Uganda. All cases occurred following sudden livestock deaths in the villages. Our findings are based on outbreak investigation which gives a better representation of the actual situation. Our article looks at the scope of the anthrax outbreak, exposures associated with illness, and evidence-based control measures. In this manuscript, we show that exposure to carcasses of animals that died suddenly was a risk factor for cutaneous anthrax in Arua District during 2015–2017. We believe the content and objectives of our work provide knowledge on epidemiology of cutaneous anthrax.

Background Schistosomiasis is a widespread public health concern in the poorest regions of the world. The principal control strategy is regular praziquantel administration to school-aged children in endemic areas. With calls for the elimination of schistosomiasis as a public health problem, expanding praziquantel delivery to all community members has been advocated. This systematic review and meta-analysis compares the impact of community-wide and child-targeted praziquantel distribution on schistosomiasis prevalence and intensity in school-aged children. Methodology/Principal findings We searched MEDLINE, Embase and Web of Science to identify papers that reported schistosome prevalence before and after praziquantel administration, either to children only or to all community members. Extracted data included Schistosoma species, drug administration strategy, number of treatment rounds, follow-up interval and prevalence and intensity before and after treatment. We used inverse variance weighted generalised linear models to examine the impact of mass versus targeted drug administration on prevalence reduction, and weighted boxplots to examine the impact on infection intensity reduction. This study is registered with PROSPERO, number CRD42018095377. In total, 34 articles were eligible for systematic review and 28 for meta-analysis. Schistosoma mansoni was reported in 20 studies; Schistosoma haematobium in 19 studies, and Schistosoma japonicum in two studies. Results of generalised linear models showed no detectable difference between mass and targeted treatment strategies on prevalence reduction in school-aged children for S. mansoni (odds ratio 0.47, 95%CI 0.13–1.68, p = 0.227) and S. haematobium (0.41, 95%CI 0.06–3.03, p = 0.358). Box plots also showed no apparent differences in intensity reduction between the two treatment strategies. Conclusions/Significance The results of this meta-analysis do not support the hypothesis that community-wide treatment is more effective than targeted treatment at reducing schistosomiasis infections in children. This may be due to the relatively small number of included studies, insufficient treatment coverage, persistent infection hotspots and unmeasured confounders. Further field-based studies comparing mass and targeted treatment are required., Author summary Schistosomiasis is a neglected tropical disease, caused by parasitic worms, that affects more than 143 million people worldwide. Chronic infections can lead to significant morbidity including kidney damage, anaemia, malnutrition, infertility and growth impairment. School-aged children between six and 15 years are often targeted for regular treatment with praziquantel in large-scale drug delivery programs, because they suffer a disproportionate burden of morbidity. On the other hand, a mass drug delivery strategy that treats all members of the community has been suggested in a move towards elimination of schistosomiasis as a public health problem. In this systematic review, we assess the impact of community-wide versus children-only praziquantel distribution in reducing schistosomiasis infections in school-aged children. We did not detect a difference between mass and targeted treatment strategies, possibly due to factors including insufficient treatment coverage and persistent sources of reinfection. Addressing these factors may assist in optimising control programs.

School health and nutrition (SHN) programmes are recognized as a significant contributor to both health and education sector goals. The school system offers an ideal platform from which to deliver basic health interventions that target the most common health conditions affecting school-age children (SAC) in low-income countries, leading to improved participation and learning outcomes. However, governments require evidence to cost, design, and implement these programmes. In Ethiopia, prevalent health conditions affecting SAC's education participation and learning outcomes include infection with soil-transmitted helminths (STHs), hunger, and malnutrition. In recognition of the multiple issues affecting the health and education of SAC, the government has taken a proactive approach, coordinating an integrated SHN programme designed to be implemented in partnership and monitored and financed through a single, integrated mechanism. The programme, known as the Enhanced School Health Initiative (ESHI), integrates three complimentary health interventions: deworming; school feeding; and provision of a water, sanitation, and hygiene (WASH) package in schools, which in delivery aim to maximize the benefits of each of the individual components. Operational research surrounding the ESHI programme includes both qualitative and quantitative analyses. Here, we present an overview of the ESHI programme and its genesis. We also introduce three additional supporting papers that provide in-depth analyses of key findings, including the baseline situational analysis, the costs, and community perceptions of the programme. The findings from ESHI provide initial evidence to develop an understanding of the related costs and synergies of integrating multiple health interventions onto a single platform. The work has translated into strengthened institutional capacity and improved cross-sectoral coordination. The government is now committed to supporting 25 million school children in Ethiopia through SHN. The ESHI model serves as a reference point for other countries looking to scale up targeted SHN interventions.

Dog rabies has been recognized from ancient times and remains widespread across the developing world with an estimated 59,000 people dying annually from the disease. In 2011 a tri-partite alliance consisting of the OIE, the WHO and the FAO committed to globally eliminating dog-mediated human rabies by 2030. Regardless of global support, the responsibility remains with local program managers to implement successful elimination programs. It is well known that vaccination programs have a high probability of successful elimination if they achieve a population-coverage of 70%. It is often quoted that reducing population turnover (typically through sterilizations) raises the probability for local elimination by maintaining herd immunity for longer. Besides this, other factors that affect rabies elimination are rarely mentioned. This paper investigates the probability for local elimination as it relates to immunity, fecundity, dog population size, infectivity (bite rates), in-migration of immune-naïve dogs, and the initial incidence. To achieve this, an individual-based, stochastic, transmission model was manipulated to create a dataset covering combinations of factors that may affect elimination. The results thereof were analysed using a logistic regression model with elimination as the dependent variable. Our results suggest that smaller dog populations, lower infectivity and lower incidence (such as when epidemics start with single introductions) strongly increased the probability for elimination at wide ranges of vaccination levels. Lower fecundity and lower in-migration had weak effects. We discuss the importance of these findings in terms of their impact and their practical application in the design of dog-mediated rabies control programs., Author summary Most guidelines for rabies control call for at least 70% vaccination coverage of dogs. This level of immunity has a very high probability for the local elimination of rabies, but it is often not an achievable ideal due to resource constraints. Campaign managers can be strategic on how they allocate their resources. Lower infectivity rates are present in areas with more restricted dog movements and have higher probabilities for elimination at lower vaccination rates. Smaller sub-populations have higher probabilities for elimination at the same vaccination coverage levels compared to larger sub-populations. Vaccinating immune corridors can divide meta-populations into smaller sub-populations that are likely to result in elimination either due to their small size or due to the local low infectivity. Areas already free of rabies require lower vaccination levels to maintain freedom compared to endemic areas. Where donors do not specifically require sterilization campaigns, funds meant for rabies control should not be diverted to sterilizations.

The Peruvian-Brazilian border is a highly endemic tegumentary leishmaniasis region in South America. The interoceanic highway is a commercial route that connects Peru and Brazil through Madre de Dios and has raised concerns about its impact on previously undisturbed areas. In order to assess leishmaniasis transmission risk along this highway, we conducted a surveillance study of the sand fly populations in this area. Sand flies were collected between 2009 and 2010 along transects at 200 m, 600 m and 1000 m from six study sites located along the highway (Iberia, La Novia, Alto Libertad, El Carmen, Florida Baja, Mazuko and Mavila) and an undisturbed area (Malinowski). Collected specimens were identified based on morphology and non-engorged females of each species were pooled and screened by kinetoplast PCR to detect natural Leishmania infections. A total of 9,023 specimens were collected belonging to 54 different Lutzomyia species including the first report of Lu. gantieri in Peru. Four species accounted for 50% of all specimens (Lutzomyia carrerai carrerai, Lu. davisi, Lu. shawi and Lu. richardwardi). El Carmen, Alto Libertad, Florida Baja and Malinowski presented higher Shannon diversity indexes (H = 2.36, 2.30, 2.17 and 2.13, respectively) than the most human disturbed sites of Mazuko and La Novia (H = 1.53 and 1.06, respectively). PCR detected 10 positive pools belonging to Lu. carrerai carrerai, Lu. yuilli yuilli, Lu. hirsuta hirsuta, Lu. (Trichophoromyia) spp., and Lu. (Lutzomyia) spp. Positive pools from 1,000 m transects had higher infectivity rates than those from 600 m and 200 m transects (9/169 = 5.3% vs 0/79 = 0% and 1/127 = 0.8%, p = 0.018). El Carmen, accounted for eight out of ten positives whereas one positive was collected in Florida Baja and Mazuko each. Our study has shown differences in sand fly diversity, abundance and species composition across and within sites. Multiple clustered Lutzomyia pools with natural Leishmania infection suggest a complex, diverse and spotty role in leishmaniasis transmission in Madre de Dios, with increased risk farther from the highway., Author summary Leishmaniasis is a neglected disease that is endemic in tropical and subtropical regions affecting up to 2 million people every year. Currently, there is concern about the effects of human-made environmental changes on the transmission of leishmaniasis due to increased exposure to the sand fly vector. In this paper, the authors explored changes in the distribution of sand flies and natural leishmania infection across communities located along the interoceanic highway that connects Peru and Brazil. The study found differences in sand fly species composition and abundance between and within sites as well as a lower sand fly diversity in communities with high human disturbance. Ten pools belonging to Lutzomyia shawi, Lu. carrerai carrerai, Lu. yuilli yuilli, Lu. hirsuta hirsuta, Lu. (Helcocyrtomyia) spp. and Lu. (Lutzomyia) spp. were found positive for Leishmania spp. The majority of these positive pools were collected from a single community and far from the highway. The information provided by this manuscript will serve as a baseline to assess the effects of human activities in the region and guide future surveillance and intervention strategies.