Mary M. Cameron

Background Visceral leishmaniasis (VL) is a debilitating disease and without treatment, a fatal disease which burdens the most impoverished communities in northeastern India. Control and ultimately, elimination of VL depends heavily on prompt case detection. However, a proportion of VL cases remain undiagnosed many months after symptom onset. Delay to diagnosis increases the chance of onward transmission, and poses a risk of resurgence in populations with waning immunity. We checked the spatial variation of delayed diagnosis of VL in Bihar, India and aimed to understand the potential driving factors of delayed diagnosis. Methods The spatial distribution of diagnostic delays was explored using a Bayesian model fit to geo-located cases using the Integrated Nested Laplace Approximation (INLA) approach, assuming days of delay as Poisson-distributed and adjusting for individual-(age, sex, HIV) and local-level (recent incidence, vector control, health facility access) characteristics. Residual variance was modelled with an explicit spatial structure. Cumulative delays were estimated under different scenarios of active case detection coverage. Findings The 4,270 cases analysed were prone to excessive delays outside existing endemic “hot spots”, beyond the focus of interventions. Cases diagnosed within recently-affected blocks and villages experienced shorter delays on average (by 13% 95% Credible Interval [2.9% - 21.7%] and 7% [1.3% - 13.1%], respectively) than those in non-recently-affected areas. Interpretation Delays to VL diagnosis when incidence is low could influence whether transmission of the disease could be interrupted or resurges. Prioritising and narrowing surveillance to high-burden areas may increase the likelihood of excessive delays in diagnosis in peripheral areas. Active surveillance driven by observed incidence may lead to missing the risk posed by as-yet-undiagnosed cases in low-endemic areas, and such surveillance could be insufficient for achieving and sustaining elimination. Funding The Bill and Melinda Gates Foundation.

: Lymphatic filariasis (LF) is a leading cause of disability due to infectious disease worldwide. The Recife Metropolitan Region (RMR) is the only remaining focus of LF in Brazil, where the parasite Wuchereria bancrofti is transmitted solely by the mosquito Culex quinquefasciatus. This study reports the results of transmission assessment surveys and molecular xenomonitoring in the city of Olinda, RMR, after nearly 15 years (2015–2016) of interventions for LF elimination. Participants were screened for W. bancrofti antigen via immunochromatographic card tests (ICT) in: 1) door-to-door surveys conducted for all children aged 5–7 years from 4 out of 17 intervention areas treated with at least five annual doses of mass drug administration (MDA), and 2) a two-stage cluster sampling survey of residents aged 5 years and older in non-MDA areas. Mosquitoes were collected via handheld aspirators in four MDA areas, differentiated by species, sex, and physiological status, pooled into groups of up to 10 blood-fed, semigravid, and gravid mosquitoes, and screened for W. bancrofti infection by real-time quantitative polymerase chain reaction (RT-qPCR). All 1,170 children from MDA areas and the entire population sample of 990 residents in non-MDA areas were ICT negative. In MDA areas, a total of 3,152 female Cx. quinquefasciatus mosquitoes in 277 households (range, 0–296 mosquitoes per house) were collected via aspiration. RT-qPCR of 233 pools of mosquitos were negative for W. bancrofti RNA; an independent reference laboratory confirmed these results. These results provide evidence that LF transmission has been halted in this setting.

A rapid, cost-effective, and simple nucleic acid isolation technique coupled with a point-of-need DNA amplification assay is a desirable goal for programmatic use. For diagnosis of Visceral Leishmaniasis (VL), Recombinase Polymerase Amplification (RPA) rapid tests for the detection of Leishmania DNA are versatile and have operational advantages over qPCR. To facilitate the delivery of the RPA test at point-of-need for VL diagnosis, we compared two rapid DNA extraction methods, SwiftDx (SX) and an in-house Boil and Spin (BS) method, coupled with RPA amplification, versus more widely used methods for DNA extraction and amplification, namely Qiagen (Q) kits and qPCR, respectively. A total of 50 confirmed VL patients and 50 controls, matched for age and gender, were recruited from Mymensingh, Bangladesh, a region highly endemic for VL. Blood samples were collected from each participant and DNA was extracted using Q, SX and BS methods. Following DNA extraction, qPCR and RPA assays were performed to detect L. donovani in downstream analysis. No significant differences in sensitivity of the RPA assay were observed between DNA extraction methods, 94.00% (95% CI: 83.45–98.75%), 90% (95% CI: 78.19–96.67%), and 88% (95% CI: 75.69–95.47%) when using Q, SX, and BS, respectively. Similarly, using qPCR, no significant differences in sensitivity were obtained when using Q or SX for DNA extraction, 94.00% (95% CI: 83.45–98.75%) and 92.00% (80.77–97.78%), respectively. It is encouraging that RPA and qPCR showed excellent agreement (k: 0.919–0.980) when different extraction methods were used and that the DNA impurities using BS had no inhibitory effect on the RPA assay. Furthermore, significantly higher DNA yields were obtained using SX and BS versus Q; however, a significantly higher parasite load was detected using qPCR when DNA was extracted using Q versus SX. Considering the cost, execution time, feasibility, and performance of RPA assay, rapid extraction methods such as the Boil and Spin technique appear to have the potential for implementation in resource-limited endemic settings. Further clinical research is warranted prior to broader application.

Background: The kala-azar elimination programme has resulted in a significant reduction in visceral leishmaniasis (VL) cases across the Indian Subcontinent. To detect any resurgence of transmission, a sensitive cost-effective surveillance system is required. Molecular xenomonitoring (MX), detection of pathogen DNA/RNA in vectors, provides a proxy of human infection in the lymphatic filariasis elimination programme. To determine whether MX can be used for VL surveillance in a low transmission setting, large numbers of the sand fly vector Phlebotomus argentipes are required. This study will determine the best method for capturing P. argentipes females for MX. Methodology/Principal findings: The field study was performed in two programmatic and two non-programmatic villages in Bihar, India. A total of 48 households (12/village) were recruited. Centers for Disease Control and Prevention light traps (CDC-LTs) were compared with Improved Prokopack (PKP) and mechanical vacuum aspirators (MVA) using standardised methods. Four 12x12 Latin squares, 576 collections, were attempted (12/house, 144/village,192/method). Molecular analyses of collections were conducted to confirm identification of P. argentipes and to detect human and Leishmania DNA. Operational factors, such as time burden, acceptance to householders and RNA preservation, were also considered. A total of 562 collections (97.7%) were completed with 6,809 sand flies captured. Females comprised 49.0% of captures, of which 1,934 (57.9%) were identified as P. argentipes. CDC-LTs collected 4.04 times more P. argentipes females than MVA and 3.62 times more than PKP (pLeishmania RNA detection and quantitation between methods were observed in experimentally infected sand flies maintained in conditions simulating field conditions. CDC-LTs were favoured by householders. Conclusions/Significance: CDC-LTs are the most useful collection tool of those tested for MX surveillance since they collected higher numbers of P. argentipes females without compromising mosquito captures or the preservation of RNA. However, capture rates are still low.

Background: The kala-azar elimination programme has resulted in a significant reduction in visceral leishmaniasis (VL) cases across the Indian Subcontinent. To detect any resurgence of transmission, a sensitive cost-effective surveillance system is required. Molecular xenomonitoring (MX), detection of pathogen DNA/RNA in vectors, provides a proxy of human infection in the lymphatic filariasis elimination programme. To determine whether MX can be used for VL surveillance in a low transmission setting, large numbers of the sand fly vector Phlebotomus argentipes are required. This study will determine the best method for capturing P. argentipes females for MX. Methodology/Principal Findings: The field study was performed in two programmatic and two non-programmatic villages in Bihar, India. A total of 48 households (12/village) were recruited. Centers for Disease Control and Prevention light traps (CDC-LTs) were compared with Improved Prokopack (PKP) and mechanical vacuum aspirators (MVA) using standardised methods. Four 12×12 Latin squares, 576 collections, were attempted (12/house, 144/village,192/method). Molecular analyses of collections were conducted to confirm identification of P. argentipes and to detect human and Leishmania DNA. Operational factors, such as time burden, acceptance to householders and RNA preservation, were also considered. A total of 562 collections (97.7%) were completed with 6,809 sand flies captured. Females comprised 49.0% of captures, of which 1,934 (57.9%) were identified as P. argentipes . CDC-LTs collected 4.04 times more P. argentipes females than MVA and 3.62 times more than PKP (pP. argentipes females without compromising mosquito captures or the preservation of RNA. However, capture rates are still low. Author Summary: Molecular xenomonitoring, screening insects for pathogen DNA/RNA, may be used for surveillance of diseases transmitted by insects. Since the proportion of insects infected with pathogens is very low in areas targeted for disease elimination, large numbers of females need to be screened. We compared three different methods for collecting Phlebotomus argentipes sand fly females, the vector of parasites causing the disease visceral leishmaniasis in the Indian subcontinent, to determine which collected the largest number of females. Other factors that may also influence selection of a particular method of collection by a disease control programme, such as the time it takes to collect and sort samples, the acceptance of householders for a particular collection method and whether RNA degradation in insect samples differed between collection methods, were also considered. Centers for Disease Control and Prevention light traps (CDC-LTs) proved to be more useful than two types of aspiration methods for collecting higher numbers of sand fly females and RNA preservation was retained. Furthermore, they took less time to install than to perform aspirations and were favoured by householders. Therefore, CDC-LTs were considered to be the most suitable collection method for molecular xenomonitoring of sand flies in India.

Bluetongue virus (BTV) and African horse sickness virus (AHSV) cause economically important diseases that are currently exotic to the United Kingdom (UK), but have significant potential for introduction and onward transmission. Given the susceptibility of animals kept in zoo collections to vector-borne diseases, a qualitative risk assessment for the introduction of BTV and AHSV to ZSL London Zoo was performed. Risk pathways for each virus were identified and assessed using published literature, animal import data and outputs from epidemiological models. Direct imports of infected animals, as well as wind-borne infected Culicoides, were considered as routes of incursion. The proximity of ongoing disease events in mainland Europe and proven capability of transmission to the UK places ZSL London Zoo at higher risk of BTV release and exposure (estimated as low to medium) than AHSV (estimated as very low to low). The recent long-range expansion of AHSV into Thailand from southern Africa highlights the need for vector competence studies of Palearctic Culicoides for AHSV to assess the risk of transmission in this region.

International laws for commercial aviation have achieved an exceptional degree of harmonisation and greatly improved passenger safety. Yet, despite much international guidance, enforceable laws for public health protection in aviation are mainly the responsibility of national authorities. As a result, public health laws may be incoherent, in conflict with other countries and/or based on disputed scientific evidence. The COVID-19 pandemic has highlighted the responsibility of airlines and regulatory authorities to protect not only air passengers but also populations in destination countries. While the greatest risk to global public health is the potential spread of disease by infected passengers or vectors, lesser-known risks include food contamination, inadequate sanitary facilities and poor air quality within the cabin. In preparedness for inevitable future disease outbreaks and pandemics, an urgent review of international law as it applies to public health in commercial aviation is needed, with greater investment in scientific research to enable more accurate and effective risk assessment and management, supported by enforceable laws and clear responsibility.

The scientific community recognizes that molecular xenomonitoring (MX) can allow infected mosquitoes to serve as a proxy for human infection in vector-borne disease surveillance, but developing reliable MX systems for programmatic use has been challenging. The primary aim of this article is to examine the available evidence to recommend how MX can best be used for various purposes. Although much of the literature published within the last 20 years focuses on using MX for lymphatic filariasis elimination, a growing body of evidence supports its use in early warning systems for emerging infectious diseases (EIDs). An MX system design must consider the goal and target (e.g. diseases targeted for elimination versus EIDs), mosquito and pathogen characteristics, and context (e.g. setting and health system). MX is currently used as a ‘supplement’ to human surveillance and will not be considered as a ‘replacement’ until the correlation between pathogen-infection rates in human and mosquito populations is better understood. Establishing such relationships may not be feasible in elimination scenarios, due to increasingly dwindling human infection prevalence after successful control, but may still be possible for EIDs and in integrated disease surveillance systems. This article is part of the theme issue ‘Novel control strategies for mosquito-borne diseases'.

Mosquito-borne diseases are an increasing global health challenge, threatening over 40% of the world's population. Despite major advances in malaria control since 2000, recent progress has stalled. Additionally, the risk of Aedes -borne arboviruses is rapidly growing, with the unprecedented spread of dengue and chikungunya viruses, outbreaks of yellow fever and the 2015 epidemic of Zika virus in Latin America. To counteract this growing problem, diverse and innovative mosquito control technologies are currently under development. Conceptually, these span an impressive spectrum of approaches, from invasive transgene cassettes with the potential to crash mosquito populations or reduce the vectorial capacity of a population, to low-cost alterations in housing design that restrict mosquito entry. This themed issue will present articles providing insight into the breadth of mosquito control research, while demonstrating the requirement for an interdisciplinary approach. The issue will highlight mosquito control technologies at varying stages of development and includes both opinion pieces and research articles with laboratory and field-based data on control strategy development. This article is part of the theme issue ‘Novel control strategies for mosquito-borne diseases'.

Significance: Methods for analyzing individual-level geo-located disease data have existed for some time, but have rarely been used to analyze endemic human diseases. Here we apply such methods to nearly a decade’s worth of uniquely detailed epidemiological data on incidence of the deadly vector-borne disease visceral leishmaniasis (VL) and its secondary condition, post–kala-azar dermal leishmaniasis (PKDL), to quantify the spread of infection around cases in space and time by inferring who infected whom, and estimate the relative contribution of different infection states to transmission. Our findings highlight the key role long diagnosis delays and PKDL play in maintaining VL transmission. This detailed characterization of the spatiotemporal transmission of VL will help inform targeting of interventions around VL and PKDL cases.