Bat Colonies Research Articles

Detection of Antimicrobial Resistance in Faecal Escherichia coli from European Free-Tailed Bats (Tadarida teniotis) in Portugal

In the last decades, the increase of antimicrobial resistance bacteria has become a concern of public health, already affecting unintentional hosts as isolated wild populations. In the present study, we evaluated the incidence of antimicrobial resistance, molecular mechanisms of resistance and virulence genes among 42 strains of Escherichia coli isolated from faecal samples from wild colonies of European free-tailed bats (Tadarida teniotis) in Portugal. High antimicrobial resistance rates for ampicillin (57.14%) and streptomycin (52.38%) were observed. Genes encoding resistance to antimicrobial agents were detected in most of the resistant isolates. The E. coli isolates were ascribed to phylogroups A (44.24%), B1 (16.67%), B2 (2.38%) and D (35.71%). The virulence gene fimA (alone or in association) was detected in 21 samples. This study shows the presence of antimicrobial resistance in commensal bacteria from wild European free-tailed bats populations. Future studies are necessary in order to investigate the main source of contamination (through the environment or the food chain) and identify how they can be contributing to the spread of those bacteria throughout the environment.

Inside or out? Cave size and landscape effects on cave-roosting bat assemblages in Brazilian Caatinga caves

AbstractCave bats have an intimate association with their roosts. Size, structural heterogeneity, and microclimatic conditions are traits of caves known to affect the structure of these assemblages. The effects of the natural and anthropogenic landscape factors around caves on the structure of these assemblages are poorly known, especially in areas with large cave clusters. We assessed the effects of cave size and surrounding landscape attributes on the richness and species composition of cave-roosting bats in 13 caves distributed in two landscapes with large cave clusters in Caatinga dry forests, Brazil. In a 1-km buffer around caves, we obtained 13 internal cave and external landscape variables. Candidate univariate models using generalized linear models were constructed and the Akaike information criterion was used for model selection. The cave size model explained richness and variance in the species composition; larger caves tended to have greater richness and assemblage composition varied depending on the cave size, hence affecting the occurrence of certain species, some of conservation concern (Natalus macrourus, Furipterus horrens). The cave connectivity model affected only the richness; caves located in denser cave clusters had higher richness likely attributed to movement of bats among caves by a more diverse array of species. Both environmental and anthropic variables affected species composition, but differently depending of the landscape context of cave location (protected versus nonprotected area). The extent these landscape variables affected the species composition was due to species-specific responses, and observed in the mean colony sizes of the species shared between the cave systems. All the landscape variables that we tested affected the structuring process of cave-roosting bats assemblages, and evidences that variables found in disturbed karstic landscapes also affect the structure of the assemblage (e.g., large colonies of vampire bats). However, the ubiquitous effect of cave size on both richness and species composition reinforces the critical importance of the roost in the life of these flying mammals.Os morcegos que se abrigam em cavernas têm uma associação muito íntima com seus abrigos. Sabe-se que as características de caverna, como o tamanho, heterogeneidade estrutural e condições microclimáticas estáveis, afetam a estrutura das assembleias dos morcegos cavernícolas. No entanto, os efeitos dos fatores externos naturais e antrópicos da paisagem em torno de cavernas na estrutura das assembleias destes morcegos são pouco conhecidos, especialmente em paisagens cársticas com grandes aglomerados de cavernas. Assim, avaliamos os efeitos do tamanho da caverna e dos atributos da paisagem circundante na riqueza e composição de espécies de morcegos em 13 cavernas distribuídas em duas paisagens de Caatinga com grandes aglomerados de cavernas no Rio Grande do Norte. Em um buffer de 1 km ao redor de cada caverna, obtivemos 13 variáveis de paisagem interna (tamanho da caverna) e externa (número de cavernas circundantes, variáveis antropogênicas e ambientais); Modelos candidatos univariados usando GLM foram construídos e o Critério de Informação de Akaike foi usado para a seleção dos modelos. O modelo de tamanho das cavernas explicou a riqueza e a variação na composição das espécies; cavernas maiores tendem a ter maior riqueza e composição da assembleia variando de acordo com o tamanho da caverna, afetando, portanto, a ocorrência de certas espécies, algumas vulneráveis (Natalus macrourus, Furipterus horrens). O modelo de conectividade da caverna afetou apenas a riqueza; cavernas localizadas em aglomerados maiores tinham maior riqueza. As variáveis ambientais e antrópicas afetaram diferentemente a composição das espécies dependendo do contexto da paisagem da localização da caverna (área protegida versus não protegida). As variáveis da paisagem afetaram a composição das espécies ocorreu de uma forma específica em algumas espécies e foi observada no tamanho médio das colônias das espécies compartilhadas entre os sistemas de cavernas. Todas as variáveis da paisagem que testamos afetaram o processo de estruturação de assembleias de morcegos cavernícolas, incluindo aquelas que são características de uma paisagem cárstica perturbada. Todas as variáveis de paisagem que testamos afetaram o processo de estruturação de assembleias de morcegos que pousam em cavernas e evidenciam que variáveis encontradas em paisagens cársticas perturbadas também afetam a estrutura da assembleia (por exemplo, grandes colônias de morcegos vampiros). No entanto, o efeito ubíquo do tamanho da caverna sobre a riqueza e a composição das espécies reforça a importância crítica do abrigo na vida desses mamíferos voadores.

The Zeyi Cave Geosite in Northern Ethiopia

Despite the high geoheritage value of caves and karsts, northern Ethiopia’s largest cave at Zeyi (13.5586°N, 39.1454°E) in the Dogu’a Tembien district has received little attention so far. We have studied its geological, geomorphic, socio-cultural and historical dimensions in a holistic way. The basal member of the Antalo Limestone, in which the Zeyi cave is located, consists of grainstone and wackestone with subordinate marly interlayers. Over a length of 364 m, the oval-shaped gallery displays stalagmites, stalactites, five columns, dissolution holes (“bell-holes”) following joints, stalagmitic floors and other concretions or speleothems. In the absence of any dating of the cave, we contrasted its elevation above the current local base level with known average incision rates of the northern Ethiopian highlands to reconstruct its age, which was calculated as at least 2 to 4 million years. The palaeo-environmental information that is archived in the Zeyi cave sediment would hence cover the Pleistocene. The graves in the sediment at the bottom of the Zeyi cave further indicate that the place could be an ancient burial site, which gives scope for archaeological research. Zeyi boosts a unique combination of abiotic, biotic and cultural components: the nineteenth c. church under the overhanging cliff; the unique cave; the speleothems, cliffs and gorges; and the cave’s bat colony which has been genetically confirmed to be composed of three syntopically roosting species. Accounting for a good balance between cave research, community-based geotourism, geoconservation and biodiversity conservation, the Zeyi cave has strong credentials to become a top geotouristic site in northern Ethiopia. However, major work needs to be done, including granting access for women and organising community-based geotourism.

PREDATION AT AN URBAN FREE-TAILED BAT (TADARIDA BRASILIENSIS) COLONY BY NIGHT HERONS

Harris County, Texas, is home to both species of North American night herons, black-crowned (Nycticorax nycticorax) and yellow-crowned (Nycticorax violacea). Both species are carnivorous, consuming a wide variety of vertebrates and invertebrates, although records of either species consuming bats have not been reported. During studies at a local colony of free-tailed bats living under the Waugh Bridge (Houston, Texas), significant feeding behavior observations of both herons were made. Both species were recorded foraging on this urban bat population, utilizing different methods for acquiring the bats as prey. Some of the behaviors are novel as well and are also discussed.

Monitoring and Characterizing Temporal Patterns of a Large Colony of Tadarida brasiliensis (Chiroptera: Molossidae) in Argentina Using Field Observations and the Weather Radar RMA1

Migratory colonies of up to thousands or millions of Brazilian free-tailed bats (Tadarida brasiliensis) are present in temperate areas of America. The monitoring of these massive colonies is crucial to know their conservation status and to evaluate the important ecosystem services that they provide. The objectives of this study were to characterize and to monitor, with an interdisciplinary approach, one of the largest bat colonies in South America, located in La Calera (Córdoba, Argentina). This study includes eight years of field observations inside of their shelter and outside when the colony emerged. Moreover, these observations were complemented with one year of weather radar detections using the Radar Meteorológico Argentino 1 (RMA1). To determine if a detection is a true or false massive emergence of bats, an algorithm was designed. We observed that this large colony of T. brasiliensis is maternal and migratory, just like others in South and North America. This colony arrives in early spring and births occur two months later, migrations occur in early autumn, meanwhile the shelter is empty or inhabited only by a small group of individuals during the cold seasons. The colony was estimated at 900,000 individuals before births occurred. The radar detection was coincident with field observations, when a simultaneous emergence was observed, as well as in the monitoring throughout the year. This represents the first study made in South America using radar technology for monitoring a bat colony. We here demonstrate that RMA1 is a powerful tool for monitoring this colony in the long term, and even to alert possible changes in permanence in time or in the number of individuals.

Timescale and colony-dependent relationships between environmental conditions and plasma oxidative markers in a long-lived bat species.

To increase the applicability and success of physiological approaches in conservation plans, conservation physiology should be based on ecologically relevant relationships between physiological markers and environmental variation that can only be obtained from wild populations. Given their integrative and multifaceted aspects, markers of oxidative status have recently been considered in conservation physiology, but still need to be validated across environmental conditions and locations. Here, we examined whether inter-annual variation in two oxidative markers, plasma antioxidant capacity and plasma hydroperoxides, followed inter-annual variation in temperature anomalies and associated vegetation changes in four colonies of long-lived greater mouse-eared bats (Myotis myotis) monitored over five consecutive years. We found that the plasma antioxidant capacity of bats decreased while plasma hydroperoxide concentrations increased with increasing temperature anomalies occurring in the two weeks before blood sampling. Moreover, the antioxidant defences of these bats reflected vegetation indices, which themselves reflected the thermal conditions experienced by bats in their foraging habitat. Variation in oxidative markers therefore appears to be due to variation in thermoregulatory costs and to indirect changes in foraging costs. Overall, these results validate the use of markers of oxidative status in conservation physiology to monitor thermal perturbations recently experienced by animals in their natural habitat. However, even though oxidative markers varied in the same direction in all four bat colonies across years, the amplitude of their response differed. If these different physiological responses reflect different performances (e.g. productivity, survival rate) between colonies, this implies that, if necessary, conservation measures may need to be applied at the local scale.

The temporal RNA virome patterns of a lesser dawn bat (Eonycteris spelaea) colony revealed by deep sequencing.

The virosphere is largely unexplored and the majority of viruses are yet to be represented in public sequence databases. Bats are rich reservoirs of viruses, including several zoonoses. In this study, high throughput sequencing (HTS) of viral RNA extracted from swabs of four body sites per bat per timepoint is used to characterize the virome through a longitudinal study of a captive colony of fruit nectar bats, species Eonycteris spelaea in Singapore. Through unbiased shotgun and target enrichment sequencing, we identify both known and previously unknown viruses of zoonotic relevance and define the population persistence and temporal patterns of viruses from families that have the capacity to jump the species barrier. To our knowledge, this is the first study that combines probe-based viral enrichment with HTS to create a viral profile from multiple swab sites on individual bats and their cohort. This work demonstrates temporal patterns of the lesser dawn bat virome, including several novel viruses. Given the known risk for bat-human zoonoses, a more complete understanding of the viral dynamics in South-eastern Asian bats has significant implications for disease prevention and control. The findings of this study will be of interest to U.S. Department of Defense personnel stationed in the Asia-Pacific region and regional public health laboratories engaged in emerging infectious disease surveillance efforts.

Faeces as a novel material to estimate lyssavirus prevalence in bat populations.

Rabies is caused by infection with a lyssavirus. Bat rabies is of concern for both public health and bat conservation. The current method for lyssavirus prevalence studies in bat populations is by oral swabbing, which is invasive for the bats, dangerous for handlers, time‐consuming and expensive. In many situations, such sampling is not feasible, and hence, our understanding of epidemiology of bat rabies is limited. Faeces are usually easy to collect from bat colonies without disturbing the bats and thus could be a practical and feasible material for lyssavirus prevalence studies. To further explore this idea, we performed virological analysis on faecal pellets and oral swabs of seven serotine bats (Eptesicus serotinus) that were positive for European bat 1 lyssavirus in the brain. We also performed immunohistochemical and virological analyses on digestive tract samples of these bats to determine potential sources of lyssavirus in the faeces. We found that lyssavirus detection by RT‐qPCR was nearly as sensitive in faecal pellets (6/7 bats positive, 86%) as in oral swabs (7/7 bats positive, 100%). The likely source of lyssavirus in the faeces was virus excreted into the oral cavity from the salivary glands (5/6 bats positive by immunohistochemistry and RT‐qPCR) or tongue (3/4 bats positive by immunohistochemistry) and swallowed with saliva. Virus could not be isolated from any of the seven faecal pellets, suggesting the lyssavirus detected in faeces is not infectious. Lyssavirus detection in the majority of faecal pellets of infected bats shows that this novel material should be further explored for lyssavirus prevalence studies in bats.

Vampire bats.

Gerald Wilkinson introduces the blood-drinking vampire bats.

Fluorescent biomarkers demonstrate prospects for spreadable vaccines to control disease transmission in wild bats.

Vaccines that autonomously transfer among individuals have been proposed as a strategy to control infectious diseases within inaccessible wildlife populations. However, rates of vaccine spread and epidemiological efficacy in real world systems remain elusive. Here, we investigated whether topical vaccines that transfer among individuals through social contacts can control vampire bat rabies, a medically and economically important zoonosis in Latin America. Field experiments in 3 Peruvian bat colonies which used fluorescent biomarkers as a proxy for the bat-to-bat transfer and ingestion of an oral vaccine revealed that vaccine transfer would increase population-level immunity up to 2.6 times beyond the same effort using conventional, non-spreadable vaccines. Mathematical models demonstrated that observed levels of vaccine transfer would reduce the probability, size, and duration of rabies outbreaks, even at low, but realistically achievable levels of vaccine application. Models further predicted that existing vaccines provide substantial advantages over culling bats, the policy currently implemented in North, Central, and South America. Linking field studies with biomarkers to mathematical models can inform how spreadable vaccines may combat pathogens of health and conservation concern prior to costly investments in vaccine design and testing.

Demographic and environmental drivers of metagenomic viral diversity in vampire bats.

Viruses infect all forms of life and play critical roles as agents of disease, drivers of biochemical cycles and sources of genetic diversity for their hosts. Our understanding of viral diversity derives primarily from comparisons among host species, precluding insight into how intraspecific variation in host ecology affects viral communities or how predictable viral communities are across populations. Here we test spatial, demographic and environmental hypotheses explaining viral richness and community composition across populations of common vampire bats, which occur in diverse habitats of North, Central and South America. We demonstrate marked variation in viral communities that was not consistently predicted by a null model of declining community similarity with increasing spatial or genetic distances separating populations. We also find no evidence that larger bat colonies host greater viral diversity. Instead, viral diversity follows an elevational gradient, is enriched by juvenile‐biased age structure, and declines with local anthropogenic food resources as measured by livestock density. Our results establish the value of linking the modern influx of metagenomic sequence data with comparative ecology, reveal that snapshot views of viral diversity are unlikely to be representative at the species level, and affirm existing ecological theories that link host ecology not only to single pathogen dynamics but also to viral communities.

Weather surveillance radar as an objective tool for monitoring bat phenology and biogeography

Bats provide a wealth of vital services to humans, such as pollinating plants and managing populations of insects that act as agricultural pests and disease vectors. Despite their direct importance, it is inherently difficult to monitor bats across large spatiotemporal scales, in part due to their nocturnal activity and long-distance, high-altitude flights. Networks of weather surveillance radars provide continuous measurements of the airspace at continental scales, revealing the abundance, distribution, behaviour, and phenology of bats aloft. Using a network of polarimetric Doppler weather radars in the USA, we demonstrate applications of this technology to monitor large bat colonies, highlight recent discoveries made using radar surveillance of bats, and discuss future prospects for extending these techniques as part of a standardised system of global bat monitoring infrastructure.

The role of past experience in development of feeding behavior in common vampire bats.

The life history strategy of common vampire bats (Desmodus rotundus) suggests that learning might play a role in development of their foraging skills. We took advantage of 12 captive births in a study colony of vampire bats to test the role of past experience in two aspects of feeding. First, we compared preferences for blood temperature in 32 wild-born vampire bats versus 11 captive-born vampire bats that had only previously fed on blood of ambient temperature or colder. We found no evidence for a preference in either group for blood presented at 4 °C versus 37 °C. Second, we tested whether captive-born vampire bats with no previous experience of feeding on live animals could successfully feed on a live chicken. Five of 12 naïve captive-born bats were able to bite the chicken and draw blood, but only one bat gained more than 5% of body mass. We were unable to reasonably compare their feeding performance with that of wild-born bats because only two of three wild-born, short-term captive bats fed on the chicken and none of the seven wild-born, long-term captive mothers attempted to feed. This unexpected lack of feeding might be due to a previously reported age-dependent neophobia. When six of the captive-born bats were released in the wild, they appeared to feed successfully because they survived for more than three consecutive nights. We suggest further tests that would better clarify the role of learning in the development of foraging in vampire bats.

Pest consumption in a vineyard system by the lesser horseshoe bat (Rhinolophus hipposideros).

Herbivorous arthropods cause immense damage in crop production annually. Consumption of these pests by insectivorous animals is of significant importance to counteract their adverse effects. Insectivorous bats are considered amongst the most voracious predators of arthropods, some of which are known crop pests. In vineyard-dominated Mediterranean agroecosystems, several crops are damaged by the attack of insect pests. In this study we aimed 1) to explore the diet and pest consumption of the lesser horseshoe bat Rhinolophus hipposideros and 2) analyse whether the composition of pest species in its diet changes throughout the season. We employed a dual-primer DNA metabarcoding analysis of DNA extracted from faeces collected in three bat colonies of a wine region in Southwestern Europe during the whole active period of most pest species. Overall, 395 arthropod prey species belonging to 11 orders were detected; lepidopterans and dipterans were the most diverse orders in terms of species. Altogether, 55 pest species were identified, 25 of which are known to cause significant agricultural damage and 8 are regarded as pests affecting grapevines. The composition of pest species in faeces changed significantly with the season, thus suggesting several periods should be sampled to assess the pest consumption by bats. As a whole, the results imply that R. hipposideros acts as a suppressor of a wide array of agricultural pests in Mediterranean agroecosystems. Therefore, management measures favouring the growth of R. hipposideros populations should be considered.

Explaining diversity patterns in dark waters – a study of aquatic caves in Yucatán, Mexico

AbstractIn the tropics, limestone caves in karstic areas are known for their unique biodiversity. However, many caves remain unstudied and little is known about underlying gradients that determine diversity and biomass in aquatic microhabitats. Here, we sampled zooplankton and benthos in a set of 12 aquatic caves, locally called closed cenotes in Yucatán, Mexico. Our aim was to explain diversity patterns and differences in biomass with particular attention for correlations between bat colony characteristics and other biota. Compared with caves that support photosynthesis, diversity was low with an average of four planktonic and two benthic species in these dark caves. Undetectable phosphorus concentrations in the water suggest this nutrient is limiting. Several associations hint at a potential link between bat abundance and functional guild composition, water quality and aquatic biota. As such, more bats were linked to higher nitrate concentrations. Yet this was not translated to higher invertebrate biomass, probably since phosphorus is limiting. Overall, the trends found in this survey suggest that bats could be important as fertilizers of the caves although mechanistic links that mediate the flux of nutrients need to be confirmed experimentally.

Rabies Outbreak in Captive Big Brown Bats (Eptesicus fuscus) Used in a White-Nose Syndrome Vaccine Trial

An outbreak of rabies occurred in a captive colony of wild-caught big brown bats (Eptesicus fuscus). Five of 27 bats exhibited signs of rabies virus infection 22-51 d after capture or 18-22 d after contact with the index case. Rabid bats showed weight loss, aggression, increased vocalization, hypersalivation, and refusal of food. Antigenic typing and virus sequencing confirmed that all five bats were infected with an identical rabies virus variant that circulates in E. fuscus in the US. Two bats with no signs of rabies virus infection were seropositive for rabies virus-neutralizing antibodies; the brains of these bats had no detectable viral proteins by the direct fluorescence antibody test. We suspect bat-to-bat transmission of rabies virus occurred among our bats because all rabies-infected bats were confined to the cage housing the index case and were infected with viruses having identical sequences of the entire rabies nucleoprotein gene. This outbreak illustrates the risk of rabies virus infection in captive bats and highlights the need for researchers using bats to assume that all wild bats could be infected with rabies virus.

Parametric Study of Dual-particle Swarm Optimisation-modified Adaptive Bats Sonar Algorithm on Multi-objective Benchmark Test Functions


 
 
 
 An integrated algorithm for solving multi-objective optimisation problems using a dual- level searching approach is presented. The proposed algorithm named as dual-particle swarm optimisation-modified adaptive bats sonar algorithm (D-PSO-MABSA) where the concept of echolocation of a colony of bats to find prey in the modified adaptive bats sonar algorithm is combined with the established particle swarm optimisation algorithm. The proposed algorithm combines the advantages of both particle swarm optimisation and modified adaptive bats sonar algorithm approach to handling the complexity of multi-objective optimisation problems. These include swarm flight attitude and swarm searching strategy. The performance of the algorithm is verified through several multi- objective optimisation benchmark test functions. The acquired results show that the proposed algorithm performs well to produce a reliable Pareto front. The proposed algorithm can thus be an effective method for solving multi-objective optimisation problems.
 
 
 

Modeling vaccination strategies to control white-nose syndrome in little brown bat colonies

Since 2006, the North American bat population has been in rapid decline due to white-nose syndrome (WNS), which is caused by an invasive fungus (Pseudogymnoascus destructans). The little brown bat (Myotis lucifugus) is the species most affected by this emerging disease. We consider how best to prevent local extinctions of this species using mathematical models. In 2017, development began on a new vaccine for WNS; we analyze the effects of implementing vaccination as a control measure. We create a Susceptible-Exposed-Infectious-Vaccinated hybrid ordinary differential equation and difference equation model informed by the phenology of little brown bats. We compare the effectiveness of annual, biennial, and one-time vaccination programs for multiple durations of immunity length. We also determine the optimal time to vaccinate, if vaccinating only once, as a function of average duration of immunity. Next, we perform a sensitivity analysis to determine the robustness of our results. Finally, we consider other possible control measures together with vaccination to determine the optimal control strategy. We find that if the vaccine offers lifelong immunity, then it will be the most effective control measure considered thus far.

The Bat Revolt in Values

Written in the wake of the strike action that has seen academics and students across more than 60 UK universities create new interstices and alliances against the marketization of Higher Education and its generalized forms of impoverishment, this parabolic essay experiments with the possibility of developing a radically immanent approach to the creation and destruction of value, and explores its implications for the concept and politics of “evaluation” at the heart of contemporary transformations in universities across the Global North. In an attempt to dramatize the interstitial spaces the strike has generated, the essay functions as a parable, relaying the complex value-ecology of the Joanina Library in Coimbra, Portugal, with its imperial and enlightened symbolisms, and its colony of bats, as a singular story that might help us envisage some elements of what might be at stake in the immanent transvaluation of the values we generate together inside and in spite of the marketizing University. This is a parable, then, to think with what is still a collective work to be done: the collective work of living, thinking, and doing otherwise in the academic ruins.

Evaluating the public health response to a mass bat exposure-Wyoming, 2017.

Mass bat exposures (MBEs) occur when multiple people are exposed to a bat or a bat colony, often over an extended period. In August 2017, a public health investigation was started in response to an MBE that occurred during May-August 2017 at a national park research station in Wyoming. We identified 176 people who had slept primarily in two lodges (Lodges A and B) at the research station, and successfully contacted 165 (93.8%) of these individuals. Risk assessments (RAs) were administered to all 165 individuals to determine degree and type of exposures to bats (e.g., biting or scratching). Exposure status for research station guests was classified as "non-exposed," "low risk" or "high risk," and counselling was provided to guide post-exposure prophylaxis (PEP) recommendations. Prior to public health notification and intervention, 19 persons made the decision to pursue PEP. The healthcare-seeking behaviours of this group were taken to represent outcomes in the absence of public health intervention. (These persons received a RA, and their risk classification was retrospectively assigned.) Approximately 1-2weeks after conducting the RAs, we conducted a follow-up survey to determine whether recommendations regarding PEP were ultimately followed. The proportion of individuals that unnecessarily pursued PEP was higher among the 19 individuals that sought health care prior to receiving the RA (p<0.00001). Among those receiving the RA first, all persons classified as high risk followed public health guidance to seek PEP treatment. Despite this, upon re-interview, only 21/79 (26.6%) of guests could accurately recall their risk classification, with most people (55.7%) overestimating their risk. Study findings demonstrate that early public health interventions such as RAs can reduce unnecessary use of PEP and that messaging used during rabies counselling should be clear.