Bats Pteropus Research Articles

Nipah virus in India: past, present and future

Nipah virus (NiV) is one of the emerging highly pathogenic virus. Like Ebola and Zika viruses, NiV too is threatening the integrity of the mankind. The family Paramyxoviruses has been traditionally associated with a group of viruses with narrow host range and typically causes outbreaks with low mortality rates. But with the emergence of highly pathogenic Hendra virus and closely related NiV, they have evolved as a cause of fatal encephalitis across broad range of vertebrate species including humans. The natural reservoir of NiV is Pteropus bat, which is apparently distributed all over the South East Asia. The bat population from North East to North West states in India have NiV antibodies which mean there is active NiV infection among Indian bats. As NiV is associated with high morbidity and mortality they pose a risk from natural outbreaks, laboratory accidents or deliberate misuse. The development of effective prevention and treatment strategies is very crucial. Preparedness, surveillance, constant vigil needs to be carried out continuously in the country. The present outbreak in India after nearly eleven years with a high case fatality rate indicate that there is a total lack of health care systems preparedness and surveillance strategy. The anthropogenic and environmental changes occurring due to rapid urbanization and massive deforestation has made India now even more vulnerable for such recurrent outbreaks. This review highlights the changing trend of the NiV outbreaks in the past and the current outbreak in India.

<b>Taking the first steps: Initial mapping of the human-wildlife interaction of the Mauritius Fruit Bat Pteropus niger (Mammalia: Chiroptera: Pteropodidae) in Mauritius by conservation organizations</b>

Interactions between people and wildlife have both positive and negative aspects. Negative interactions, commonly termed human-wildlife conflict (HWC), have increased in recent decades due to a number of factors including difficulties in identifying and communicating the complexities of stakeholder values and positions over wildlife and its management. Here, we present the perceptions of two conservation organizations on the landscape of HWC involving the threatened Mauritius Fruit Bat Pteropus niger, Kerr 1792 in Mauritius, including damage to fruit crops and controversial government culls in 2015 and 2016. Participants identified 18 stakeholders in the conflict varying in importance and influence, examined where and how hostility is manifested, and delineated both perceived and real costs of the conflict. Additionally, 13 environmental and 17 social risk factors associated with the conflict were categorized, along with potential policy and management options for mitigation. We argue that initial in-house workshops are advantageous in understanding conservation conflicts before extending dialogue with other stakeholders.

Bat Astrovirus in Mozambique

Astroviruses (AstVs) are responsible for infection of a large diversity of mammalian and avian species, including bats, aquatic birds, livestock and humans. We investigated AstVs circulation in bats in Mozambique and Mayotte, a small island in the Comoros Archipelago located between east Africa and Madagascar. Biological material was collected from 338 bats and tested for the presence of the AstV RNA-dependent RNA-polymerase gene with a pan-AstV semi-nested polymerase chain reaction assay. None of the 79 samples obtained from Mayotte bats (Pteropus seychellensis comorensis and Chaerephon pusillus) tested positive; however, 20.1% of bats sampled in Mozambique shed AstVs at the time of sampling and significant interspecific variation in the proportion of positive bats was detected. Many AstVs sequences obtained from a given bat species clustered in different phylogenetic lineages, while others seem to reflect some level of host-virus association, but also with AstVs previously reported from Malagasy bats. Our findings support active circulation of a large diversity of AstVs in bats in the western Indian Ocean islands, including the southeastern African coast, and highlight the need for more detailed assessment of its risk of zoonotic transmission to human populations.

Emergence of Nipah Virus: Need More R&D and Public Health Infrastructure

Emergence of Nipah Virus: Need More R&D and Public Health Infrastructure

A bat on the brink? A range-wide survey of the Critically Endangered Livingstone's fruit bat Pteropus livingstonii

AbstractThe Livingstone's fruit bat Pteropus livingstonii is endemic to the small islands of Anjouan and Mohéli in the Comoros archipelago, Indian Ocean. The species is under threat from anthropogenic pressure on the little that remains of its forest habitat, now restricted to the islands’ upper elevations and steepest slopes. We report the results of the most comprehensive survey of this species to date, and present recommendations for ongoing field conservation efforts and monitoring. Morning counts were conducted at roost sites in the wet and dry seasons during 2011–2013. Habitat structure around the roosting sites was characterized and roost numbers compared, to investigate the potential effect of habitat loss and degradation. We estimate the population to comprise c. 1,260 individuals distributed across 21 roosts on the two islands. All occupied roosting sites were restricted to a narrow altitudinal range, and roosting populations in agroforestry areas were smaller than those found in degraded and undisturbed forest. Only one of the 16 roosts on Anjouan was found in undisturbed, old-growth forest with no nearby signs of clearance for agriculture or landslides following tree-felling upslope. Following a suspected severe population decline as a result of widespread and long-term forest loss Livingstone's fruit bat has been recategorized as Critically Endangered on the IUCN Red List.

Phenotypic and functional characterization of the major lymphocyte populations in the fruit-eating bat Pteropus alecto.

The unique ability of bats to act as reservoir for viruses that are highly pathogenic to humans suggests unique properties and functional characteristics of their immune system. However, the lack of bat specific reagents, in particular antibodies, has limited our knowledge of bat’s immunity. Using cross-reactive antibodies, we report the phenotypic and functional characterization of T cell subsets, B and NK cells in the fruit-eating bat Pteropus alecto. Our findings indicate the predominance of CD8+ T cells in the spleen from wild-caught bats that may reflect either the presence of viruses in this organ or predominance of this cell subset at steady state. Instead majority of T cells in circulation, lymph nodes and bone marrow (BM) were CD4+ subsets. Interestingly, 40% of spleen T cells expressed constitutively IL-17, IL-22 and TGF-β mRNA, which may indicate a strong bias towards the Th17 and regulatory T cell subsets. Furthermore, the unexpected high number of T cells in bats BM could suggest an important role in T cell development. Finally, mitogenic stimulation induced proliferation and production of effector molecules by bats immune cells. This work contributes to a better understanding of bat’s immunity, opening up new perspectives of therapeutic interventions for humans.

KOMPOSISI KIMIA DAGING SEGAR DAN SIFAT ORGANOLEPTIK KELELAWAR OLAHAN

CHEMICAL COMPOSITION AND ORGANOLEPTIC CHARACTERISTICS OF PROCESSING BAT (Pteropus alecto) FRESH MEAT. Study was done to evaluate chemical composition and preference degree of the consumers on the processing bat meat compared with beef, chicken and cakalang fish using steam cooking method, flavor cooking and spicy cooking method. Chemical composition was analyzed by proximate analysis using procedures of the Association of Official Analytical Chemists. The organoleptic test was done by hedonic test. Data of the hedonic test were analyzed using Kruskal-Wallis method. To evaluate data distribution of the panelist respons on the sample test, the data were using plot graphic box. The hedonic scales were using 1 to 7. The variables used were including color, taste, flavor, tenderness and general acceptance. Results of proximate analyses of bat meat based on fresh meat indicated that water content was 67.21 percents and protein was 20.48 percents. Based on dry matter, the protein contents of bat meat, pork, chicken, and cakalang fish were 48.97 percents, 69.08 percents, 67.14 percents, and 69.41 percents, respectively. The water contents were 5.76 percents, 9.92 percents, 8.27 percents, and 9.90 percents, repsectively; the calcium (Ca) contents were also 10.62 percents, 1.09 percents, 1.36 percents, 1.38 percents, respectively; while Phosphor (P) contents were 1.46 percents, 0.69 percents, 0.66 percents, and 0.72 percents, respectively. Variance analysis for organoleptic test showed that meat type with different processing affected significantly on taste, color, flavor, tenderness and general preference of bat meat, beef meat, chicken meat and cakalang fish by steam cooking, chicken flavor, and spicy cooking methods. Conclusion, bat meat contained protein relatively lower than that in pork meat, chicken meat, and cakalang fish, but mineral contents were relative higher. Results of organoleptic test showed that preference degree of processing meat were higher on using spicy flavor. The specific test on taste, color, flavor, tenderness and general responsed showed relatively the same responses by the panelists. Key word: Chemical composition, organoleptic test, processing bat meat

Factors explaining the dispersal of Hendra virus disease in South-East Queensland

Hendra virus (HeV) was first described in 1994 following the outbreak of a new disease fatally affecting horses and humans in South-east Queensland. The disease kills 70% of the infected horses and under some circumstances the virus is spread to humans who have had close contact with the infected horses. Fruit bats (Pteropus spp.) commonly known as flying foxes have been identified as the natural host of the virus. A preliminary analysis of the incidents and the flying fox roosting sites revealed a strong relationship between the temporary and seasonal roosting sites in the South-east Queensland. The aim of this paper is to determine the potential factors that can explain the dispersal of HeV incidents in the study area. Based on the preliminary results, a further analysis was done on the roosting sites by considering factors such as the species of flying foxes, foraging range and pregnancy period. Spatial autocorrelation (Global Moran’s I) revealed significant clustering of P. alecto and P. scapulatus species. Kernel density estimation analysis helped in identifying a strong relationship between P. alecto & P. scapulatus species density and the outbreak events in the study area. Buffer analysis established an initial relationship between P. alecto & P. poliocephalus species birth periods and the incidents. Ordinary least-squares regression identified P. poliocephalus species as statistically significant at a global context across the study area. Geographically weighted regression analysis was performed to study the local spatial variations of the explanatory variables. P. alecto & P. poliocephalus species exhibited a significant positive relationship in most of the regions where as pregnancy period variable exhibited a significant negative relationship to the HeV incidents in the study area.

Molecular characterization of Nipah virus from Pteropus hypomelanus in Southern Thailand.

BackgroundNipah virus (NiV) first emerged in Malaysia in 1998, with two bat species (Pteropus hypomelanus and P. vampyrus) as the putative natural reservoirs. In 2002, NiV IgG antibodies were detected in these species from Thailand, but viral RNA could not be detected for strain characterization. Two strains of NiV (Malaysia and Bangladesh) have been found in P. lylei in central Thailand, although Bangladesh strain, the causative strain for the outbreak in Bangladesh since 2001, was dominant. To understand the diversity of NiV in Thailand, this study identified NiV strain, using molecular characterizations, from P. hypomelanus in southern Thailand.FindingsPooled bat urine specimens were collected from plastic sheet underneath bat roosts in April 2010, and then monthly from December 2010 to May 2011 at an island in southern Thailand. Five in 184 specimens were positive for NiV, using duplex nested RT-PCR assay on partial nucleocapsid fragment (357 bp). Whole sequences of nucleocapsid gene from four bats were characterized. All 5 partial fragments and 4 whole nucleocapsid genes formed a monophyletic with NiV-MY.ConclusionsOur study showed that P. hypomelanus in southern Thailand and from Malaysia, a bordering country, harbored similar NiV. This finding indicates that NiV is not limited to central Thailand or P. lylei species, and it may be a source of inter-species transmission. This indicates a higher potential for a widespread NiV outbreak in Thailand. NiV surveillance in Pteropus bats, the major natural reservoirs, should be conducted continuously in countries or regions with high susceptibility to outbreaks.

Mapping the risk of Nipah virus spillover into human populations in South and Southeast Asia.

Nipah virus (NiV) is a significant emerging zoonotic pathogen given its wide geographic distribution, and the severe morbidity and high mortality that accompanies infection. Moreover, the layered landscape epidemiology surrounding spillover from reservoir host species to humans is ill-defined. Identifying landscape features that contribute to NiV spillover would likely prove helpful in preventing emergence in human populations. Using an inhomogeneous Poisson model, this study investigated the role of vegetation cover, the human footprint (HFP) and reservoir Pteropus bat distribution to identify the spatial dependence of spillover and map risk across South and Southeast Asia. The spatial model identified HFP (RR=1.08; 95% CI 1.05-1.11) and bat distribution (RR=19.44; 95% CI 1.92-196.7) as significant predictors of NiV risk, while vegetation cover was not significant after accounting for HFP and the presence of Pteropus bats. These findings further inform the landscape epidemiology of NiV and suggest specific conduits for spillover in the landscape. However, more detailed field studies will be required to validate these results.

Identification of a novel Afipia species isolated from an Indian flying fox.

An old world fruit bat Pteropus giganteus, held in captivity and suffering from necrosis of its wing digits, failed to respond to antibiotic therapy and succumbed to the infection. Samples submitted to the National Centre for Foreign Animal Disease were tested for viral infection. Vero E6 cells exhibited minor but unique cytopathic effects on second blind passage, and full CPE by passage four. Utilizing an unbiased random amplification technique from cell culture supernatant, we identified a bacterium belonging to the Bradyrhizobiaceae. Purification of cell culture supernatant on TY media revealed a slow growing bacterial isolate. In this study using electron microscopy, 16S rRNA gene analysis and whole genome sequencing, we identify a novel bacterial species associated with the site of infection belonging to the genus Afipia. This genus of bacteria is very diverse, with only a limited number of species characterized. Afipia felis, previously described as the etiological agent to cause cat scratch disease, and Afipia septicemium, most recently shown to cause disease in humans, highlight the potential for members of this genus to form a branch of opportunistic pathogens within the Bradyrhizobiaceae. Increased utilization of next generation sequencing and genomics will aid in classifying additional members of this intriguing bacterial genera.

Serological evidence of henipavirus exposure in cattle, goats and pigs in Bangladesh.

BackgroundNipah virus (NiV) is an emerging disease that causes severe encephalitis and respiratory illness in humans. Pigs were identified as an intermediate host for NiV transmission in Malaysia. In Bangladesh, NiV has caused recognized human outbreaks since 2001 and three outbreak investigations identified an epidemiological association between close contact with sick or dead animals and human illness.MethodologyWe examined cattle and goats reared around Pteropus bat roosts in human NiV outbreak areas. We also tested pig sera collected under another study focused on Japanese encephalitis.Principal FindingsWe detected antibodies against NiV glycoprotein in 26 (6.5%) cattle, 17 (4.3%) goats and 138 (44.2%) pigs by a Luminex-based multiplexed microsphere assay; however, these antibodies did not neutralize NiV. Cattle and goats with NiVsG antibodies were more likely to have a history of feeding on fruits partially eaten by bats or birds (PR = 3.1, 95% CI 1.6–5.7) and drinking palmyra palm juice (PR = 3.9, 95% CI 1.5–10.2).ConclusionsThis difference in test results may be due to the exposure of animals to one or more novel viruses with antigenic similarity to NiV. Further research may identify a novel organism of public health importance.

Scanning electron microscopy study of guard hair of three Indian pteropodid bats

A comparative study on the hair microstructure of pteropodid bats Pteropus giganteus , Rousettus leschenaulti , and Cynopterus sphinx was performed using scanning electron microscope. Hair samples were taken from the dorsal, ventral, and neck regions. Among the three pteropodids examined, an imbricate and tightly appressed hair type was observed in P. giganteus , while a coronal type of cuticle with margins diverging from the shaft was observed in R. leschenaulti and C. sphinx . The coronal cuticles of R. leschenaulti and C. sphinx had relatively thinner scale widths than the imbricate cuticles of P. giganteus . There was a significant difference in hair length, scale lengths and scale widths, and scale indices among the three species of fruit bats. However, there was no significant difference in hair length and scale index of male and female R. leschenaulti as well as in hair length and scale indices of male and female C. sphinx . The presence of bell-shaped coronal cuticle and wide angular scale margin in R. leschenaulti and C. sphinx reveal their role as active pollinators. The hair patterns, hair length, scale length and width confirm the presence of species-specific characteristics of pteropodid bats and can be used for species identification.

Chapter 32 - Henipavirus encephalitis

Henipaviruses are new zoonotic paramyxoviruses that can cause fatal encephalitis in humans. They are comprised of Hendra virus, which was first discovered in Australia in 1994 in association with infected horses, and Nipah virus, which was first discovered in Malaysia in 1999 during a fatal outbreak involving more than 350 patients with 40% mortality, in association with infected pigs. The mechanism of zoonotic transfer is via droplet inhalation or direct contact with secretion or excretion from infected animals. The natural host reservoir is identified as fruit bats of the Pteropus species based on the discovery of neutralizing antibody, with horses and pigs acting as amplifying hosts. The pathologic hallmark of Henipavirus infection is disseminated microinfarctions as a result of vasculitis-induced thrombosis. The diagnosis of Henipavirus infection is based on a history of contact with sick animals (mammals), evidence of encephalitis and/or pneumonia, with serologic evidence of infection using enzyme-linked immunosorbent assay testing or polymerase chain reaction. Magnetic resonance imaging of the brain was a sensitive and specific diagnostic tool. Electroencephalogram was non-specific but correlated well with the severity of disease. Ribavirin has been used in the treatment of Henipavirus encephalitis, with a 36% reduction in mortality during the Malaysian outbreak. Relapsed and late-onset encephalitis has been reported in less than 10% of survivors. In view of the wide geographic distribution of Pteropus bats, Henipavirus remains an important cause of zoonotic fatal encephalitis worldwide.

Lack of population genetic structure and host specificity in the bat fly, Cyclopodia horsfieldi, across species of Pteropus bats in Southeast Asia

BackgroundPopulation-level studies of parasites have the potential to elucidate patterns of host movement and cross-species interactions that are not evident from host genealogy alone. Bat flies are obligate and generally host-specific blood-feeding parasites of bats. Old-World flies in the family Nycteribiidae are entirely wingless and depend on their hosts for long-distance dispersal; their population genetics has been unstudied to date.MethodsWe collected a total of 125 bat flies from three Pteropus species (Pteropus vampyrus, P. hypomelanus, and P. lylei) from eight localities in Malaysia, Cambodia, and Vietnam. We identified specimens morphologically and then sequenced three mitochondrial DNA gene fragments (CoI, CoII, cytB; 1744 basepairs total) from a subset of 45 bat flies. We measured genetic diversity, molecular variance, and population genetic subdivision (FST), and used phylogenetic and haplotype network analyses to quantify parasite genetic structure across host species and localities.ResultsAll flies were identified as Cyclopodia horsfieldi with the exception of two individuals of Eucampsipoda sundaica. Low levels of population genetic structure were detected between populations of Cyclopodia horsfieldi from across a wide geographic range (~1000 km), and tests for isolation by distance were rejected. AMOVA results support a lack of geographic and host-specific population structure, with molecular variance primarily partitioned within populations. Pairwise FST values from flies collected from island populations of Pteropus hypomelanus in East and West Peninsular Malaysia supported predictions based on previous studies of host genetic structure.ConclusionsThe lack of population genetic structure and morphological variation observed in Cyclopodia horsfieldi is most likely due to frequent contact between flying fox species and subsequent high levels of parasite gene flow. Specifically, we suggest that Pteropus vampyrus may facilitate movement of bat flies between the three Pteropus species in the region. We demonstrate the utility of parasite genetics as an additional layer of information to measure host movement and interspecific host contact. These approaches may have wide implications for understanding zoonotic, epizootic, and enzootic disease dynamics. Bat flies may play a role as vectors of disease in bats, and their competence as vectors of bacterial and/or viral pathogens is in need of further investigation.

The pandemic potential of Nipah virus

Nipah virus, a paramyxovirus whose wildlife reservoir is Pteropus bats, was first discovered in a large outbreak of acute encephalitis in Malaysia in 1998 among persons who had contact with sick pigs. Apparently, one or more pigs was infected from bats, and the virus then spread efficiently from pig to pig, then from pigs to people. Nipah virus outbreaks have been recognized nearly every year in Bangladesh since 2001 and occasionally in neighboring India. Outbreaks in Bangladesh and India have been characterized by frequent person-to-person transmission and the death of over 70% of infected people. Characteristics of Nipah virus that increase its risk of becoming a global pandemic include: humans are already susceptible; many strains are capable of limited person-to-person transmission; as an RNA virus, it has an exceptionally high rate of mutation: and that if a human-adapted strain were to infect communities in South Asia, high population densities and global interconnectedness would rapidly spread the infection. Appropriate steps to estimate and manage this risk include studies to explore the molecular and genetic basis of respiratory transmission of henipaviruses, improved surveillance for human infections, support from high-income countries to reduce the risk of person-to-person transmission of infectious agents in low-income health care settings, and consideration of vaccination in communities at ongoing risk of exposure to the secretions and excretions of Pteropus bats.

Duration of Maternal Antibodies against Canine Distemper Virus and Hendra Virus in Pteropid Bats

Old World frugivorous bats have been identified as natural hosts for emerging zoonotic viruses of significant public health concern, including henipaviruses (Nipah and Hendra virus), Ebola virus, and Marburg virus. Epidemiological studies of these viruses in bats often utilize serology to describe viral dynamics, with particular attention paid to juveniles, whose birth increases the overall susceptibility of the population to a viral outbreak once maternal immunity wanes. However, little is understood about bat immunology, including the duration of maternal antibodies in neonates. Understanding duration of maternally derived immunity is critical for characterizing viral dynamics in bat populations, which may help assess the risk of spillover to humans. We conducted two separate studies of pregnant Pteropus bat species and their offspring to measure the half-life and duration of antibodies to 1) canine distemper virus antigen in vaccinated captive Pteropus hypomelanus; and 2) Hendra virus in wild-caught, naturally infected Pteropus alecto. Both of these pteropid bat species are known reservoirs for henipaviruses. We found that in both species, antibodies were transferred from dam to pup. In P. hypomelanus pups, titers against CDV waned over a mean period of 228.6 days (95% CI: 185.4–271.8) and had a mean terminal phase half-life of 96.0 days (CI 95%: 30.7–299.7). In P. alecto pups, antibodies waned over 255.13 days (95% CI: 221.0–289.3) and had a mean terminal phase half-life of 52.24 days (CI 95%: 33.76–80.83). Each species showed a duration of transferred maternal immunity of between 7.5 and 8.5 months, which was longer than has been previously estimated. These data will allow for more accurate interpretation of age-related Henipavirus serological data collected from wild pteropid bats.

Detection of Nipah Virus RNA in Fruit Bat (Pteropus giganteus) from India

The study deals with the survey of different bat populations (Pteropus giganteus, Cynopterus sphinx, and Megaderma lyra) in India for highly pathogenic Nipah virus (NiV), Reston Ebola virus, and Marburg virus. Bats (n = 140) from two states in India (Maharashtra and West Bengal) were tested for IgG (serum samples) against these viruses and for virus RNAs. Only NiV RNA was detected in a liver homogenate of P. giganteus captured in Myanaguri, West Bengal. Partial sequence analysis of nucleocapsid, glycoprotein, fusion, and phosphoprotein genes showed similarity with the NiV sequences from earlier outbreaks in India. A serum sample of this bat was also positive by enzyme-linked immunosorbent assay for NiV-specific IgG. This is the first report on confirmation of Nipah viral RNA in Pteropus bat from India and suggests the possible role of this species in transmission of NiV in India.

Proteolytic Processing of Hendra Virus Fusion Protein in Cells of the Bat Reservoir Host

Hendra and Nipah are recently emerged, zoonotic paramyxoviruses with high mortality rates. While the majority of paramyxovirus fusion (F) proteins are proteolytically activated by the host protease furin, Henipavirus F proteins utilize the protease cathepsin L. The primary natural reservoir of Henipaviruses is fruit bats. Interestingly, fruit bats act as reservoirs for SARS‐like coronavirus and Ebola virus, both of which use cathepsins during their life cycles. We analyzed proteolytic processing of viral glycoproteins in recently generated cell lines from the fruit bats Pteropus alecto and Rousettus aegyptus. Both the cathepsin‐dependent Hendra virus F and the furin‐dependent parainfluenza virus 5 F proteins were properly cleaved, suggesting that furin and cathepsin‐like proteases function in these cells. Syncytia formation was observed with expression of either F protein, confirming that proteolytic activation resulted in fusogenic molecules. Addition of cathepsin L inhibitors inhibited Hendra F, but not PIV5 F cleavage in all cell types. There results demonstrate that proteolytic activation of Henipavirus F proteins by cathepsin is not due to lack of furin‐like proteases in Pteropus bat cells.This work was supported by NIAID/NIH grant R01AI051517 and NIH grant #U54 AI057157 from the Southeastern Regional Center of Excellence for Emerging Infections and Biodefense.

Hendra virus detection using Loop-Mediated Isothermal Amplification

Hendra virus (HeV) is a zoonotic paramyxovirus endemic in Australian Pteropus bats (fruit bats or flying foxes). Although bats appear to be unaffected by the virus, HeV can spread from fruit bats to horses, causing severe disease. Human infection results from close contact with the blood, body fluids and tissues of infected horses. HeV is a biosecurity level 4 (BSL-4) pathogen, with a high case-fatality rate in humans and horses. Current assays for HeV detection require complex instrumentation and are generally time consuming. The aim of this study was to develop a Loop-Mediated Isothermal Amplification (LAMP) assay to detect nucleic acid from all known HeV strains in horses without the requirement for complex laboratory equipment. A LAMP assay targeting a conserved region of the HeV P-gene was combined with a Lateral Flow Device (LFD) for detection of amplified product. All HeV isolates, the original HeV isolated in 1994 as well as the most recent isolates from 2011 were detected. Analytical sensitivity and specificity of the HeV-LAMP assay was equal to a TaqMan assay developed previously. Significantly, these assays detected HeV in horses before clinical signs were observed. The combined LAMP-LFD procedure is a sensitive method suitable for HeV diagnosis in a resource-limited situation or where rapid test results are critical.