Estimating the Size of Populations at High Risk of Malaria in 2 Operational Districts in Cambodia: Household-Based Survey.

BACKGROUND: Cambodia is targeting the elimination of malaria by 2025. The last remaining pockets of malaria in Cambodia are concentrated among populations exposed to forested areas, but the size of these populations is not well understood. To plan for the procurement and distribution of vector-control tools, chemoprophylaxis, and other commodities for malaria prevention and surveillance, robust estimates of the population at greatest risk are required. OBJECTIVE: This study aims to estimate the number of forest-exposed individuals residing in Cambodia's highest-burden operational districts (ODs) in 2 provinces with active malaria transmission. METHODS: In April 2023, a multistage, in-person survey was conducted among residents in the 2 ODs in Cambodia with the highest malaria burden: Sen Monorom in Mondulkiri province and Phnom Srouch in Kampong Speu province. In each OD, 10 villages were randomly selected, and 35 households were randomly selected from each village. To estimate the number of individuals at high risk of malaria-defined as residing within 1 km of a forest or traveling at least once per week to the forest-respondents were asked about the distance from their household to the nearest forested area, and their travel patterns to forested areas. To account for mobility (ie, to avoid double-counting), respondents also provided information on overnight stays at other households in the selected villages in the past month. In the 4 selected villages in Sen Monorom OD where Project BITE forest packs (an intervention in the larger research program) had been distributed prior to the survey, respondents were also asked questions to determine if they had received such a pack, to develop smaller scale "multiplier method" estimates of at-risk individuals in each of those villages. RESULTS: In Sen Monorom, 138 households and 872 individuals were enrolled in the survey, and in Phnom Srouch, 163 households and 844 individuals were enrolled. The estimated percentage of female householders was 49.7% (852/1716) across both ODs; the median age was 22 (IQR 12-37) years in Sen Monorom and 24.5 (IQR 16.0-40.5) years in Phnom Srouch (total age range 3-86). Based on mobility-adjusted survey estimates alone, 32% (280/706; 95% CI 19.9-47.2) of residents in Sen Monorom (an estimated 12,133-20,135 individuals) and 36% (68/198; 95% CI 24.5-45.5) of residents in Phnom Srouch (an estimated 1717-2203 individuals), met risk criteria for forest exposure. Between 125 and 186 individuals were estimated to be at risk in each of the 4 villages where the multiplier method could be applied. CONCLUSIONS: This study provides estimates of the number of individuals potentially at high risk for malaria infection due to forest exposure in 2 ODs in Cambodia. These estimates can support planning for malaria control and elimination efforts. The straightforward methods of household surveys and multipliers should be feasible for many national malaria control programs.

Field evaluation of a volatile pyrethroid spatial repellent and etofenprox treated clothing for outdoor protection against forest malaria vectors in Cambodia.

Cambodia's goal to eliminate malaria by 2025 is challenged by persistent transmission in forest and forest fringe areas, where people are exposed to Anopheles mosquito bites during the day and night. Volatile pyrethroid spatial repellents (VPSRs) and insecticide-treated clothing (ITC) could address these gaps. This study evaluated the outdoor application of one passive transfluthrin-based VPSR, four etofenprox-ITCs paired with a picaridin topical repellent, and a combination of VPSR and ITC against wild Anopheles landing in Cambodia. A 7 × 7 Latin-square study was conducted over 49 collection nights in temporary open structures in Mondulkiri Province. All interventions substantially reduced Anopheles landing, with protective efficacy ranging from 61 to 95%. Mathematical modeling showed significant reductions in vectoral capacity, especially with the combined ITC and VPSR and VPSR alone, albeit with decreased effectiveness over time. These interventions have the potential to reduce outdoor and daytime Anopheles biting, offering valuable contributions to malaria elimination efforts in Cambodia and the Greater Mekong Subregion, contingent upon achieving effective coverage and adherence.

Inference for entomological semi-field experiments: Fitting a mathematical model assessing personal and community protection of vector-control interventions.

The effectiveness of vector-control tools is often assessed by experiments as a reduction in mosquito landings using human landing catches (HLCs). However, HLCs alone only quantify a single characteristic and therefore do not provide information on the overall impacts of the intervention product. Using data from a recent semi-field study which used time-stratified HLCs, aspiration of non-landing mosquitoes, and blood feeding, we suggest a Bayesian inference approach for fitting such data to a stochastic model. This model considers both personal protection, through a reduction in biting, and community protection, from mosquito mortality and disarming (prolonged inhibition of blood feeding). Parameter estimates are then used to predict the reduction of vectorial capacity induced by etofenpox-treated clothing, picaridin topical repellents, transfluthrin spatial repellents and metofluthrin spatial repellents, as well as combined interventions for Plasmodium falciparum malaria in Anopleles minimus. Overall, all interventions had both personal and community effects, preventing biting and killing or disarming mosquitoes. This led to large estimated reductions in the vectorial capacity, with substantial impact even at low coverage. As the interventions aged, fewer mosquitoes were killed; however the impact of some interventions changed from killing to disarming mosquitoes. Overall, this inference method allows for additional modes of action, rather than just reduction in biting, to be parameterised and highlights the tools assessed as promising malaria interventions.

Applying the COM-B behaviour change model to a pilot study delivering volatile pyrethroid spatial repellents and insecticide-treated clothing to forest-exposed populations in Mondulkiri Province, Cambodia.

BACKGROUND: Southeast Asia is making tremendous progress towards their 2030 malaria elimination goal but needs new interventions to stop forest malaria. This study trials two new vector control tools, a volatile pyrethroid spatial repellent (VPSR) and insecticide-treated clothing (ITC), amongst forest-exposed populations in Mondulkiri Province Cambodia to inform their potential use for eliminating forest malaria. METHODS: 21 forest-exposed individuals were given a questionnaire on their perceptions of malaria and preventive practices used, after which they trialed two products sequentially. Clothes was treated with ITC by the study team. Mixed methods were used to understand their experience, attitudes, and preferences regarding the products trialed. Quantitative data was summarized and qualitative insights were analysed using thematic analysis, applying the Capability, Opportunity, and Motivation Behaviour Change (COM-B) model and Behaviour Change Wheel Framework to identify intervention functions to support tailored product rollout amongst these populations. RESULTS: Study participants reported a need for protection from mosquito bites in outdoor and forest-exposed settings and perceived both products trialed to be effective for this purpose. The VPSR product was preferred when travel was not required, whereas ITC was preferred for ease of use when going to the forest, especially in rainy conditions. COM-B analysis identified that key enablers for use of both products included their perceived efficacy and ease of use, which required no skill or preparation. For barriers to use, the odour of ITC was sometimes perceived as being toxic, as well as its inability to protect uncovered skin from mosquito bites, while the perceived usefulness of the VPSR product trialed was limited by its water sensitivity in rainy forest settings. Intervention components to encourage appropriate and sustained use of these products include education about how to use these products and what to expect, persuasion to use them from community leaders and targeted channels, and enablement to facilitate convenient and affordable access. CONCLUSION: The rollout of VPSRs and ITC amongst forest-exposed populations can be useful for eliminating malaria in Southeast Asia. Study findings can be applied to increase product uptake among forest exposed populations in Cambodia, while manufacturers can aim to develop products that are rainproof, easy to use in forest settings, and have favourable odour profiles to target users.

A semi-field evaluation in Thailand of the use of human landing catches (HLC) versus human-baited double net trap (HDN) for assessing the impact of a volatile pyrethroid spatial repellent and pyrethroid-treated clothing on Anopheles minimus landing.

BACKGROUND: The mosquito landing rate measured by human landing catches (HLC) is the conventional endpoint used to evaluate the impact of vector control interventions on human-vector exposure. Non-exposure based alternatives to the HLC are desirable to minimize the risk of accidental mosquito bites. One such alternative is the human-baited double net trap (HDN), but the estimated personal protection of interventions using the HDN has not been compared to the efficacy estimated using HLC. This semi-field study in Sai Yok District, Kanchanaburi Province, Thailand, evaluates the performance of the HLC and the HDN for estimating the effect on Anopheles minimus landing rates of two intervention types characterized by contrasting modes of action, a volatile pyrethroid spatial repellent (VSPR) and insecticide-treated clothing (ITC). METHODS: Two experiments to evaluate the protective efficacy of (1) a VPSR and (2) ITC, were performed. A block randomized cross-over design over 32 nights was carried out with both the HLC or HDN. Eight replicates per combination of collection method and intervention or control arm were conducted. For each replicate, 100 An. minimus were released and were collected for 6 h. The odds ratio (OR) of the released An. minimus mosquitoes landing in the intervention compared to the control arm was estimated using logistic regression, including collection method, treatment, and experimental day as fixed effects. RESULTS: For the VPSR, the protective efficacy was similar for the two methods: 99.3%, 95% CI (99.5-99.0) when measured by HLC, and 100% (100, Inf) when measured by HDN where no mosquitoes were caught (interaction test p = 0.99). For the ITC, the protective efficacy was 70% (60-77%) measured by HLC but there was no evidence of protection when measured by HDN [4% increase (15-27%)] (interaction test p < 0.001). CONCLUSIONS: Interactions between mosquitoes, bite prevention tools and the sampling method may impact the estimated intervention protective efficacy. Consequently, the sampling method must be considered when evaluating these interventions. The HDN is a valid alternative trapping method (relative to the HLC) for evaluating the impact of bite prevention methods that affect mosquito behaviour at a distance (e.g. VPSR), but not for interventions that operate through tarsal contact (e.g., ITC).

Applying the COM-B behavior model to inform the delivery of spatial repellents and insecticide-treated clothing among forest exposed populations in Mondulkiri Province, Cambodia.

BACKGROUND: Southeast Asia is making tremendous progress towards their 2030 malaria elimination goal but needs new interventions to stop forest malaria. This study trials two new vector control tools, a volatile pyrethroid spatial repellent (VSPR) and insecticide-treated clothing (ITC), amongst forest-exposed populations in Mondulkiri Province Cambodia to inform their potential use for eliminating forest malaria. METHODS: 21 forest-exposed individuals were given a questionnaire on their perceptions of malaria and preventive practices used, after which they trialed two products sequentially. Mixed methods were used to understand their experience, attitudes, and preferences regarding the products trialed. Quantitative data was summarized and qualitative insights were analyzed using thematic analysis, applying the Capability, Opportunity, Motivation Behavior Change (COM-B) model and Behavior Change Wheel Framework to identify intervention functions to support tailored product rollout amongst these populations. RESULTS: Study participants reported a need for protection from mosquito bites in outdoor and forest-exposed settings and perceived both products trialed to be effective for this purpose. The VPSR product was preferred when travel was not required, whereas ITC was preferred for ease of use when going to the forest, especially in rainy conditions. COM-B analysis identified that key enablers for use of both products included their perceived efficacy and ease of use, which required no skill or preparation. For barriers to use, the odor of ITC was sometimes perceived as being toxic, as well as its inability to protect uncovered skin from mosquito bites, while the perceived usefulness of the VPSR product trialed was limited by its water sensitivity in rainy forest settings. Intervention components to encourage appropriate and sustained use of these products include education about how to use these products and what to expect, persuasion to use them from community leaders and targeted ads, and enablement to guarantee access. CONCLUSION: The rollout of VPSRs and ITC amongst forest-exposed populations can be useful for eliminating malaria in Southeast Asia. Study findings can be applied to increase product uptake in Cambodia, while research efforts can aim to develop products that are rainproof, easy to use in forest settings, and have favorable odor profiles to target users.

Perceptions and preparedness of senior medical students about antimicrobial stewardship programs: are we providing adequate training to future prescribers?

BACKGROUND: Senior medical students will become one of the key partners in antimicrobial stewardship efforts in the future, yet the level of education and their perceptions toward this topic are not well documented in China. RESEARCH DESIGN AND METHODS: We conducted a cross-sectional, anonymous, online survey between December 2021 and February 2022. The students came from six universities of all five provinces/autonomous regions in northwest China. Students completed the survey by using WeChat. RESULTS: More than half of students agreed/strongly agreed that antimicrobials are overused (53.1%) and that antimicrobial resistance is a significant problem nationally (50.2%). Most of the respondents (70%) were interested in learning more about antimicrobials. Around 60% of the respondents thought they were well prepared for future use of antimicrobials. Only 30% of the respondents were familiar with the term 'Antimicrobial Stewardship,' but 80.7% were interested in taking part in an antimicrobial stewardship program training. More than half of the senior medical students thought that courses in antibacterial is suitable for second and third academic years. CONCLUSIONS: It is therefore suggested to provide specific curriculum and strengthen training of antimicrobial use for medical students in the future, as well as more rotation practice in infectious diseases related departments.

Preferences of pregnant women toward a future maternal Group B Streptococcus vaccine in China: A cross-sectional survey with a discrete choice experiment.

Group B streptococcus (GBS) is a major cause of neonatal death worldwide. A GBS vaccine for pregnant women is under development and is expected to be available in the near future. The perceptions and preferences of pregnant women in China of GBS vaccines has not been investigated, and this study aimed to investigate pregnant women's awareness of GBS and their potential preferences for the GBS vaccine. A discrete choice experiment was conducted among pregnant women in hospitals from Shaanxi, Hunan, and Zhejiang provinces located in Western, Central, and Eastern China, respectively. A conditional logit model was used to analyze the data and calculate willingness to pay values and choice probabilities of different GBS vaccine programs. A total of 354 pregnant women were included in the final analysis, 45.8% of whom were willing to receive a GBS vaccine if it were licensed. Vaccine safety was the most important attribute of a future vaccine, while cost was the least important attribute. Compared with no vaccination, pregnant women had a strong preference for future GBS vaccination (ASC = 1.267, p < .001). Pregnant women's decisions were highly influenced by those of other pregnant women. Improving the safety, efficacy, and vaccination rate of the GBS vaccine in China is of great significance for future GBS vaccine development and vaccination. Compared to other variable options, the cost of a GBS vaccine was of the least importance among pregnant women in mainland China. These findings can inform public health policy decisions related to GBS vaccination in China.

Targeted genomic sequencing with probe capture for discovery and surveillance of coronaviruses in bats.

Public health emergencies like SARS, MERS, and COVID-19 have prioritized surveillance of zoonotic coronaviruses, resulting in extensive genomic characterization of coronavirus diversity in bats. Sequencing viral genomes directly from animal specimens remains a laboratory challenge, however, and most bat coronaviruses have been characterized solely by PCR amplification of small regions from the best-conserved gene. This has resulted in limited phylogenetic resolution and left viral genetic factors relevant to threat assessment undescribed. In this study, we evaluated whether a technique called hybridization probe capture can achieve more extensive genome recovery from surveillance specimens. Using a custom panel of 20,000 probes, we captured and sequenced coronavirus genomic material in 21 swab specimens collected from bats in the Democratic Republic of the Congo. For 15 of these specimens, probe capture recovered more genome sequence than had been previously generated with standard amplicon sequencing protocols, providing a median 6.1-fold improvement (ranging up to 69.1-fold). Probe capture data also identified five novel alpha- and betacoronaviruses in these specimens, and their full genomes were recovered with additional deep sequencing. Based on these experiences, we discuss how probe capture could be effectively operationalized alongside other sequencing technologies for high-throughput, genomics-based discovery and surveillance of bat coronaviruses.

Predicting the potential for zoonotic transmission and host associations for novel viruses.

Host-virus associations have co-evolved under ecological and evolutionary selection pressures that shape cross-species transmission and spillover to humans. Observed virus-host associations provide relevant context for newly discovered wildlife viruses to assess knowledge gaps in host-range and estimate pathways for potential human infection. Using models to predict virus-host networks, we predicted the likelihood of humans as hosts for 513 newly discovered viruses detected by large-scale wildlife surveillance at high-risk animal-human interfaces in Africa, Asia, and Latin America. Predictions indicated that novel coronaviruses are likely to infect a greater number of host species than viruses from other families. Our models further characterize novel viruses through prioritization scores and directly inform surveillance targets to identify host ranges for newly discovered viruses.

Wildlife in Cameroon harbor diverse coronaviruses, including many closely related to human coronavirus 229E.

Zoonotic spillover of animal viruses into human populations is a continuous and increasing public health risk. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) highlights the global impact of emergence. Considering the history and diversity of coronaviruses (CoVs), especially in bats, SARS-CoV-2 will likely not be the last to spillover from animals into human populations. We sampled and tested wildlife in the Central African country Cameroon to determine which CoVs are circulating and how they relate to previously detected human and animal CoVs. We collected animal and ecological data at sampling locations and used family-level consensus PCR combined with amplicon sequencing for virus detection. Between 2003 and 2018, samples were collected from 6,580 animals of several different orders. CoV RNA was detected in 175 bats, a civet, and a shrew. The CoV RNAs detected in the bats represented 17 different genetic clusters, coinciding with alpha (n = 8) and beta (n = 9) CoVs. Sequences resembling human CoV-229E (HCoV-229E) were found in 40 Hipposideridae bats. Phylogenetic analyses place the human-derived HCoV-229E isolates closest to those from camels in terms of the S and N genes but closest to isolates from bats for the envelope, membrane, and RNA-dependent RNA polymerase genes. The CoV RNA positivity rate in bats varied significantly (P < 0.001) between the wet (8.2 per cent) and dry seasons (4.5 per cent). Most sampled species accordingly had a wet season high and dry season low, while for some the opposite was found. Eight of the suspected CoV species of which we detected RNA appear to be entirely novel CoV species, which suggests that CoV diversity in African wildlife is still rather poorly understood. The detection of multiple different variants of HCoV-229E-like viruses supports the bat reservoir hypothesis for this virus, with the phylogenetic results casting some doubt on camels as an intermediate host. The findings also support the previously proposed influence of ecological factors on CoV circulation, indicating a high level of underlying complexity to the viral ecology. These results indicate the importance of investing in surveillance activities among wild animals to detect all potential threats as well as sentinel surveillance among exposed humans to determine emerging threats.

The Impact of a National Stewardship Policy on the Usage Patterns of Key Monitoring Drugs in a Tertiary Teaching Hospital: An Interrupted Time Series Analysis.

Background: The management of Key Monitoring Drugs has become one of important aspects to control the growth of pharmaceutical expenditures in China. The first batch of the China National Key Monitoring Drugs (NKMDs) policy was released in July 2019. However, little is known about the impact of the national stewardship on the trends of NKMDs prescribing practice in hospitals, especially in the Northwestern China. Methods: We collected 8-years of monthly NKMDs usage data from a tertiary hospital between 2014 and 2021. A segmented regression model of interrupted time series (ITS) analysis was used to evaluate the Defined Daily Doses (DDDs) and spending trends of ten NMKDs in the hospital throughout the study period. The pre-implementation period was from January 2014 to November 2019 and the post-implementation period was from December 2019 to June 2021. Results: Prior to the implementation of the NKMDs policy, there was an increasing trend both in DDDs and spending for 8 of 10 NKMDs. The interventions managed by clinical pharmacists after the implementation of the national stewardship policy led to a significant decreasing trend of DDDs in the 19 months following implementation, of 430 fewer DDDs per month in total, compared to the pre-implementation period (p < 0.001). A similar decrease in spending was seen in the post-implementation period, with a trend of $4,682 less total spending on medications in those months compared to the pre-implementation trend (p = 0.003). There was a significant decrease in both monthly DDDs and spending for 6 of the 10 medications in the post-implementation period, while there was a significant increased trend both in monthly DDDs and spending on 1 medication in that period. Conclusion: Using ITS analysis, the total DDDs and spending on 10 NKMDs in this hospital indicated sustained reductions over 19 months after multidimensional interventions under the implementation of the national policy guidance. The national stewardship policy could therefore be considered an effective strategy. Additional comprehensive policies should be introduced to further improve the rational use of NKMDs.

Risk perceptions and behaviors of actors in the wild animal value chain in Kinshasa, Democratic Republic of Congo.

In the Democratic Republic of Congo (DRC) which contains the greatest area of the second largest rainforest on Earth, people have long been connected to the forest for subsistence and livelihood from wild animals and bushmeat. This qualitative study sought to characterize the bushmeat movement-from hunting wild animals to market sale-and the roles of participants in the animal value chain, as well as their beliefs surrounding zoonotic disease and occupational risk. Actors in in eight bushmeat markets and two ports in Kinshasa, DRC completed semi-structured interviews between 2016 and 2018 in which they expressed belief in transmission of illness from domestic animals to humans, but not from wild animals to humans. Wild animals were viewed as pure and natural, in contrast to domestic animals which were considered tainted by human interference. Participants reported cutting themselves during the process of butchering yet did not consider butchering bushmeat to be a risky activity. Instead, they adopted safety practices learned over time from butchering experts and taught themselves how to butcher in a fashion that reduced the frequency of cutting. In general, butcherers rejected the idea of personal protective equipment use. Port markets were identified as important access points for meat coming from the Congo river and plane transport was identified as important for fresh and live meat coming from Équateur province. Most participants reported having heard about Ebola, but their mistrust in government messaging privileged a word-of-mouth story of witchcraft to be propagated about Ebola's origins. It is critical to better understand how public health messaging about outbreaks can successfully reach high risk communities, and to develop creative risk mitigation strategies for populations in regular contact with animal blood and body fluids. In this paper, we offer suggestions for formal and informal trusted channels through which health messages surrounding zoonotic risk could be conveyed to high-risk populations in Kinshasa.

Coronavirus surveillance in wildlife from two Congo basin countries detects RNA of multiple species circulating in bats and rodents.

Coronaviruses play an important role as pathogens of humans and animals, and the emergence of epidemics like SARS, MERS and COVID-19 is closely linked to zoonotic transmission events primarily from wild animals. Bats have been found to be an important source of coronaviruses with some of them having the potential to infect humans, with other animals serving as intermediate or alternate hosts or reservoirs. Host diversity may be an important contributor to viral diversity and thus the potential for zoonotic events. To date, limited research has been done in Africa on this topic, in particular in the Congo Basin despite frequent contact between humans and wildlife in this region. We sampled and, using consensus coronavirus PCR-primers, tested 3,561 wild animals for coronavirus RNA. The focus was on bats (38%), rodents (38%), and primates (23%) that posed an elevated risk for contact with people, and we found coronavirus RNA in 121 animals, of which all but two were bats. Depending on the taxonomic family, bats were significantly more likely to be coronavirus RNA-positive when sampled either in the wet (Pteropodidae and Rhinolophidae) or dry season (Hipposideridae, Miniopteridae, Molossidae, and Vespertilionidae). The detected RNA sequences correspond to 15 alpha- and 6 betacoronaviruses, with some of them being very similar (>95% nucleotide identities) to known coronaviruses and others being more unique and potentially representing novel viruses. In seven of the bats, we detected RNA most closely related to sequences of the human common cold coronaviruses 229E or NL63 (>80% nucleotide identities). The findings highlight the potential for coronavirus spillover, especially in regions with a high diversity of bats and close human contact, and reinforces the need for ongoing surveillance.

Antimicrobial Stewardship Programs in Northwest China: A Cross-Sectional Survey of Perceptions, Involvement, and Perceived Barriers Among Hospital Pharmacists.

Background: Antimicrobial stewardship (AMS) is a key prevention strategy in addressing the global concern of increasing antimicrobial resistance (AMR). Pharmacists are one of the integral members of AMS hospital teams around the world. Toward reducing AMR, a major strategy in China is to improve the capacity and participation of pharmacists in the AMS framework. However, little is known about how hospital pharmacists perceive their position and participation in AMS work, and the barriers to this work in China, especially in the Northwest region. Methods: Region this work describes a cross-sectional, anonymous, online survey study. Hospital pharmacists from five provinces/autonomous regions in northwest China were invited to participate in June and July 2020. Participants completed the survey by using WeChat, a popular social application in China. We purposefully distributed the questionnaire link and QR code to hospital pharmacists through the hospital antimicrobial resistance surveillance network, hospital antimicrobial consumption surveillance network, provincial and city pharmaceutical associations, and hospital pharmacist WeChat groups. Results: Out of 1032 respondents, 93.1% believed that AMS programs promote the judicial prescribing of antimicrobials, 95.5% strongly agreed that AMS could reduce the widespread use of antimicrobials, and 92.3% believed that AMS could improve medical services. Pharmacists were most likely to be involved in AMS through reviewing prescriptions of antimicrobials, intervening in inappropriate prescriptions, and providing feedback on antimicrobial prescriptions and medical orders. Barriers to participating in AMS included workload (59.5% of respondents), ineffective communication between pharmacists and doctors (57.7%), and inadequate knowledge of AMS (47.0%). Differences in responses were found between the five surveyed provinces. A significant association was found between median involvement scores and gender, age, education, level of superiority, experience, and type of hospital (p < 0.05). Conclusion: Pharmacists perceived that AMS programs are important, but that their involvement in related activities is limited in all provinces. Further studies and strategies should consider how to overcome the identified barriers to optimize the participation of pharmacists in AMS programs.

Evaluation of bat adenoviruses suggests co-evolution and host roosting behaviour as drivers for diversity.

Adenoviruses (AdVs) are diverse pathogens of humans and animals, with several dozen bat AdVs already identified. Considering that over 100 human AdVs are known, and the huge diversity of bat species, many bat AdVs likely remain undiscovered. To learn more about AdV prevalence, diversity and evolution, we sampled and tested bats in Cameroon using several PCR assays for viral and host DNA. AdV DNA was detected in 14 % of the 671 sampled animals belonging to 37 different bat species. There was a correlation between species roosting in larger groups and AdV DNA detection. The detected AdV DNA belonged to between 28 and 44 different, mostly previously unknown, mastadenovirus species. The novel isolates are phylogenetically diverse and while some cluster with known viruses, others appear to form divergent new clusters. The phylogenetic tree of novel and previously known bat AdVs does not mirror that of the various host species, but does contain structures consistent with a degree of virus-host co-evolution. Given that closely related isolates were found in different host species, it seems likely that at least some bat AdVs have jumped species barriers, probably in the more recent past; however, the tree is also consistent with such events having taken place throughout bat AdV evolution. AdV diversity was highest in bat species roosting in large groups. The study significantly increased the diversity of AdVs known to be harboured by bats, and suggests that host behaviours, such as roosting size, may be what limits some AdVs to one species rather than an inability of AdVs to infect other related hosts.

Market characteristics and zoonotic disease risk perception in Cameroon bushmeat markets.

Behavioral practices are one of the key factors facilitating zoonotic disease transmission, especially in individuals who have frequent contact with wild animals, yet practices of those who work and live in high-risk animal-human interfaces, such as wild animal 'bushmeat' markets in the Congo Basin are not well documented in the social, health and medical sciences. This region, where hunting, butchering, and consumption of wild animal meat is frequent, represents a hotspot for disease emergence, and has experienced zoonotic disease spillover events, traced back to close human-animal contact with bats and non-human primates. Using a One Health approach, we conducted wildlife surveillance, human behavioral research, and concurrent human and animal biological sampling to identify and characterize factors associated with zoonotic disease emergence and transmission. Research was conducted through the USAID Emerging Pandemic Threats program between 2010 and 2019 including qualitative studies of bushmeat markets, with selected study sites prioritized based on proximity to bushmeat markets. Sites included two hospitals where we conducted surveillance of individuals with syndromes of acute febrile illness, community sites where we enrolled actors of the animal value chain (ie. hunters, middlemen, transporters), and bushmeat markets, where we enrolled bushmeat vendors, butchers, market managers, cleaners, and shoppers. Mixed methods research was undertaken at these sites and included investigation of bushmeat market dynamics through observational research, focus group discussions, quantitative questionnaires, and interviews. Participants were asked about their risk perception of zoonotic disease transmission and specific activities related to bushmeat trade, local market conditions, and regulations on bushmeat trade in Cameroon. Risks associated with blood contact and animal infection were not well understood by most market actors. As bushmeat markets are an important disease interface, as seen with CoVID19, risk mitigation measures in markets and bushmeat alternative strategies are discussed.

Human Interactions with Bat Populations in Bombali, Sierra Leone.

Human contact with bats has been epidemiologically linked to several of the most recent Ebola outbreaks, including the 2014 West Africa epidemic and the 2007 Luebo, Democratic Republic of the Congo, outbreak. While fruit bats remain the likely primary reservoir for Ebola virus (Zaire ebolavirus), recent wildlife surveillance efforts have identified a new species of ebolavirus (Bombali ebolavirus) in microchiropteran insect-eating bats in West and East Africa. Given the role of bats as potential Ebola reservoirs and sources of spillover into human populations, it is critically important to understand the circumstances and behaviors that bring human populations into close contact with bats. This study explores two sites in Bombali, Sierra Leone, where human populations have had close contact with microchiropteran bats via household infestations and fruit bats by hunting practices. Through interviews and focus groups, we identify the knowledge, beliefs, perceptions, and behaviors that may potentially protect or expose individuals to zoonotic spillover through direct and indirect contact with bats. We also describe how this research was used to develop a risk reduction and outreach tool for living safely with bats.

Serology and Behavioral Perspectives on Ebola Virus Disease Among Bushmeat Vendors in Equateur, Democratic Republic of the Congo, After the 2018 Outbreak.

After the 2018 Ebola outbreak in Equateur Province, Democratic Republic of the Congo, we conducted behavioral interviews and collected samples from bushmeat vendors and primates in Mbandaka to test for evidence of Ebola virus exposure. Although participants indicated being aware of Ebola, they did not consider themselves at occupational risk for infection. We found antibodies against Zaire ebolavirus in one participant despite no reported history of disease or contact with infected individuals. Our data underline concerns of possible subclinical or undiagnosed Ebola virus infections and the importance and challenges of risk communication to populations who are occupationally exposed to bushmeat. Following the 2018 Ebola outbreak in Equateur Province, Democratic Republic of the Congo, bushmeat vendors interviewed in Mbandaka indicated being aware of Ebola, but did not consider themselves at occupational risk. Antibodies against Zaire ebolavirus were detected in one participant.