Newly discovered viruses of flying foxes.

Flying foxes have been the focus of research into three newly described viruses from the order Mononegavirales, namely Hendra virus (HeV), Menangle virus and Australian Bat Lyssavirus (ABL). Early investigations indicate that flying foxes are the reservoir host for these viruses. In 1994, two outbreaks of a new zoonotic disease affecting horses and humans occurred in Queensland. The virus which was found to be responsible was called equine morbillivirus (EMV) and has since been renamed HeV. Investigation into the reservoir of HeV has produced evidence that antibodies capable of neutralising HeV have only been detected in flying foxes. Over 20% of flying foxes in eastern Australia have been identified as being seropositive. Additionally six species of flying foxes in Papua New Guinea have tested positive for antibodies to HeV. In 1996 a virus from the family Paramyxoviridae was isolated from the uterine fluid of a female flying fox. Sequencing of 10000 of the 18000 base pairs (bp) has shown that the sequence is identical to the HeV sequence. As part of investigations into HeV, a virus was isolated from a juvenile flying fox which presented with neurological signs in 1996. This virus was characterised as belonging to the family Rhabdoviridae, and was named ABL. Since then four flying fox species and one insectivorous species have tested positive for ABL. The third virus to be detected in flying foxes is Menangle virus, belonging to the family Paramyxoviridae. This virus was responsible for a zoonotic disease affecting pigs and humans in New South Wales in 1997. Antibodies capable of neutralising Menangle virus, were detected in flying foxes.     

亨得拉病毒核蛋白基因(N)在大肠杆菌的高效表达与反应原性鉴定

从引进的含亨得拉病毒（Hendra virus，HeV）基因的克隆质粒中扩增得到该病毒核蛋白基因（N），将其克隆到pET-28a（＋）原核表达栽体，转化大肠杆菌后成功地表达了N蛋白，SDS-PAGE电泳分析结果表明目的基因在JM109（DE3）大肠杆菌菌株中得到了良好的表达，目的蛋白量占菌体总蛋白的14．6％。Western-blotting检测表明，表达的蛋白可以被兔抗亨得拉全病毒阳性血清识别，表明该蛋白具有良好的反应性。该研究结果可进一步应用于诊断试剂和单克隆抗体的制备以及流行病学调查，以防范该病在我国的流行。     

Avoiding bites and scratches? Understanding the public health implication of human–bat interactions in Ghana

Zoonotic pathogens cause an estimated 70% of emerging and re‐emerging infectious diseases in humans, affecting various aspects of human development on a global scale. The significance of bats as a source of emerging infectious diseases is being progressively appreciated. This study was undertaken post‐Ebola virus disease in West Africa and assessed the public health implications of human–bat interactions by exploring the reasons for contact between humans and bats, as well as reported actions taken upon experiencing bat bites or scratches. The paper highlights the nuances of human–bat interactions, stressing zoonotic disease risk awareness as well as the sources of information. The study used questionnaires to solicit information from 788 respondents in five communities with significant bat populations. We show that bat consumption was one of the main reasons for human–bat interactions. More men across the various communities ate bat meat. Only a small number of respondents (4.4%) reported being bitten by a bat, and 6.1% had been scratched by a bat. More than 21% had come into direct contact with bat blood. An even lower number went to the hospital after been bitten or scratched by bats. There was little knowledge on post‐exposure management. The most common places human–bat interactions occurred were at home and on farms. Seventy‐three per cent of the respondents believed that bats carried diseases, with Ebola virus disease being the most mentioned. Respondents indicated that the way they interacted with bats had not changed, even though they believed bats carried diseases and 46% stated that they had not changed the way they interacted with bats over the last two years. Apart from providing information on avoiding bites and scratches, a more holistic framework is needed to reduce human–bat interactions. The paper recommends a comprehensive and coordinated approach to optimizing an effective response to a potential bat‐borne zoonotic disease spillover.     

Animal models of henipavirus infection: A review

Hendra virus (HeV) and Nipah virus (NiV) form a separate genus Henipavirus within the family Paramyxoviridae, and are classified as biosafety level four pathogens due to their high case fatality rate following human infection and because of the lack of effective vaccines or therapy. Both viruses emerged from their natural reservoir during the last decade of the 20th century, causing severe disease in humans, horses and swine, and infecting a number of other mammalian species. The current review summarises current published data relating to experimental infection of small and large animals, including the natural reservoir species, the Pteropus bat, with HeV or NiV. Susceptibility to infection and virus distribution in the individual species is discussed, along with the pathogenesis, pathological changes, and potential routes of transmission.     

Hendra and Nipah virus infections.

The most important clinical and pathological manifestation of Hendra virus infection in horses and humans is that of severe interstitial pneumonia caused by viral infection of small blood vessels. The virus is also capable of causing nervous disease. Hendra virus is not contagious in horses and is spread by close contact with body fluids, such as froth from infected lungs. Diagnosis should be based on the laboratory examination of blood, lung, kidney, spleen, and, if nervous signs are present, also of the brain. Evidence of infection with the more recently identified and related Nipah virus was found in the brain of one horse in which there was inflammation of the meningeal blood vessels. Fruit bats, especially Pteropus s., have been incriminated as the natural and reservoir hosts of both Hendra and Nipah viruses.     

No Evidence of Hendra Virus Infection in the Australian Flying‐fox Ectoparasite Genus Cyclopodia

Hendra virus (HeV) causes potentially fatal respiratory and/or neurological disease in both horses and humans. Although Australian flying‐foxes of the genus Pteropus have been identified as reservoir hosts, the precise mechanism of HeV transmission has yet to be elucidated. To date, there has been limited investigation into the role of haematophagous insects as vectors of HeV. This mode of transmission is particularly relevant because Australian flying‐foxes host the bat‐specific blood‐feeding ectoparasites of the genus Cyclopodia (Diptera: Nycteribiidae), also known as bat flies. Using molecular detection methods, we screened for HeV RNA in 183 bat flies collected from flying‐foxes inhabiting a roost in Boonah, Queensland, Australia. It was subsequently demonstrated that during the study period, Pteropus alecto in this roost had a HeV RNA prevalence between 2 and 15% (95% CI [1, 6] to [8, 26], respectively). We found no evidence of HeV in any bat flies tested, including 10 bat flies collected from P. alecto in which we detected HeV RNA. Our negative findings are consistent with previous findings and provide additional evidence that bat flies do not play a primary role in HeV transmission.     

Hendra Virus Infection in Horses: A Review on Emerging Mystery Paramyxovirus

Hendra virus (HeV) is a zoonotic paramyxovirus which causes acute and deadly infection in horses (Equus caballus). It is a rare and unmanaged emerging viral infection in horses which is harbored by bats of the genus Pteropus (Australian flying foxes or fruit bats). The virus is pleomorphic in shape and its genome contains nonsegmented negative-stranded RNA with 18234 nucleotides in length. The virus is transmitted from flying foxes to horses, horse to horse, and horse to humans. Human-to-human transmission of HeV infection is not reported yet. The infection of HeV in horses is highly variable and shows broad range of signs and lesions including distinct respiratory and neurological disorders. Currently, there are no specific antiviral drugs available for the treatment of HeV infection in horses. Vaccination is considered as prime option to prevent HeV infection in horses. A subunit vaccine, called as “Equivac HeV vaccine” has been approved recently for preventing this viral infection in horses. In addition, a plethora of common preventive strategies could help restrict the inter- and intra-species transmission of HeV. Considering the scanty but severe fatality cases of this mystery virus as well as lack of proper attention by veterinary scientists, this review article spotlights not only on the clinical signs, transmission, epidemiology, biology, pathogenesis, and diagnosis of HeV but also the preventive managements of this uncommon infection in horses by vaccination and other precautious strategies.     

Zoonotic origin and transmission of Middle East respiratory syndrome coronavirus in the UAE

Since the emergence of Middle East respiratory syndrome coronavirus (MERS‐CoV) in 2012, there have been a number of clusters of human‐to‐human transmission. These cases of human‐to‐human transmission involve close contact and have occurred primarily in healthcare settings, and they are suspected to result from repeated zoonotic introductions. In this study, we sequenced whole MERS‐CoV genomes directly from respiratory samples collected from 23 confirmed MERS cases in the United Arab Emirates (UAE). These samples included cases from three nosocomial and three household clusters. The sequences were analysed for changes and relatedness with regard to the collected epidemiological data and other available MERS‐CoV genomic data. Sequence analysis supports the epidemiological data within the clusters, and further, suggests that these clusters emerged independently. To understand how and when these clusters emerged, respiratory samples were taken from dromedary camels, a known host of MERS‐CoV, in the same geographic regions as the human clusters. Middle East respiratory syndrome coronavirus genomes from six virus‐positive animals were sequenced, and these genomes were nearly identical to those found in human patients from corresponding regions. These data demonstrate a genetic link for each of these clusters to a camel and support the hypothesis that human MERS‐CoV diversity results from multiple zoonotic introductions.     

Contact Variables for Exposure to Avian Influenza H5N1 Virus at the Human–Animal Interface

Although the highly pathogenic avian influenza H5N1 virus continues to cause infections in both avian and human populations, the specific zoonotic risk factors remain poorly understood. This review summarizes available evidence regarding types of contact associated with transmission of H5N1 virus at the human–animal interface. A systematic search of the published literature revealed five analytical studies and 15 case reports describing avian influenza transmission from animals to humans for further review. Risk factors identified in analytical studies were compared, and World Health Organization‐confirmed cases, identified in case reports, were classified according to type of contact reported using a standardized algorithm. Although cases were primarily associated with direct contact with sick/unexpectedly dead birds, some cases reported only indirect contact with birds or contaminated environments or contact with apparently healthy birds. Specific types of contacts or activities leading to exposure could not be determined from data available in the publications reviewed. These results support previous reports that direct contact with sick birds is not the only means of human exposure to avian influenza H5N1 virus. To target public health measures and disease awareness messaging for reducing the risk of zoonotic infection with avian influenza H5N1 virus, the specific types of contacts and activities leading to transmission need to be further understood. The role of environmental virus persistence, shedding of virus by asymptomatic poultry and disease pathophysiology in different avian species relative to human zoonotic risk, as well as specific modes of zoonotic transmission, should be determined.     

Risk factors for disease associated with influenza virus infections during three epidemics in horses

OBJECTIVE: To identify risk factors associated with respiratory tract disease in horses during 3 epidemics caused by influenza virus infections. DESIGN: Cross-sectional and prospective longitudinal observational studies. ANIMALS: 1,163 horses stabled at a Thoroughbred racetrack. PROCEDURES: Investigations were conducted during a 3-year period. An epidemic of respiratory tract disease caused by influenza virus infections was identified in each year. Routine observations and physical examinations were used to classify horses' disease status. Data were analyzed to identify factors associated with development of disease. RESULTS: Results were quite similar among the epidemics. Concentrations of serum antibodies against influenza virus and age were strongly associated with risk of disease; young horses and those with low antibody concentrations had the highest risk of disease. Calculation of population attributable fractions suggested that respiratory tract disease would have been prevented in 25% of affected horses of all horses had high serum antibody concentrations prior to exposure. However, recent history of vaccination was not associated with reduction in disease risk. Exercise ponies had greater risk of disease than racehorses, which was likely attributable to frequent horse-to-horse contact. CONCLUSIONS AND CLINICAL RELEVANCE: Particular attention should be paid to young horses, those with low serum antibody concentrations, and horses that have frequent contact with other horses when designing and implementing control programs for respiratory tract disease caused by influenza virus infections. It appears that control programs should not rely on the efficacy of commercial vaccines to substantially reduce the risk of disease caused by influenza virus infections.     

Human–Dromedary Camel Interactions and the Risk of Acquiring Zoonotic Middle East Respiratory Syndrome Coronavirus Infection

Middle East respiratory syndrome coronavirus (MERS‐CoV) cases without documented contact with another human MERS‐CoV case make up 61% (517/853) of all reported cases. These primary cases are of particular interest for understanding the source(s) and route(s) of transmission and for designing long‐term disease control measures. Dromedary camels are the only animal species for which there is convincing evidence that it is a host species for MERS‐CoV and hence a potential source of human infections. However, only a small proportion of the primary cases have reported contact with camels. Other possible sources and vehicles of infection include food‐borne transmission through consumption of unpasteurized camel milk and raw meat, medicinal use of camel urine and zoonotic transmission from other species. There are critical knowledge gaps around this new disease which can only be closed through traditional field epidemiological investigations and studies designed to test hypothesis regarding sources of infection and risk factors for disease. Since the 1960s, there has been a radical change in dromedary camel farming practices in the Arabian Peninsula with an intensification of the production and a concentration of the production around cities. It is possible that the recent intensification of camel herding in the Arabian Peninsula has increased the virus' reproductive number and attack rate in camel herds while the ‘urbanization’ of camel herding increased the frequency of zoonotic ‘spillover’ infections from camels to humans. It is reasonable to assume, although difficult to measure, that the sensitivity of public health surveillance to detect previously unknown diseases is lower in East Africa than in Saudi Arabia and that sporadic human cases may have gone undetected there.     

Experimental inoculation study indicates swine as a potential host for Hendra virus

Hendra virus (HeV) is a zoonotic virus from the family Paramyxoviridae causing fatal disease in humans and horses. Five-week-old Landrace pigs and 5-month-old Gottingen minipigs were inoculated with approximately 10⁷ plaque forming units per animal. In addition to fever and depression exhibited in all infected pigs, one of the two Landrace pigs developed respiratory signs at 5 days post-inoculation (dpi) and one of the Gottingen minipigs developed respiratory signs at 5 dpi and mild neurological signs at 7 dpi. Virus was detected in all infected pigs at 2–5 dpi from oral, nasal, and rectal swabs and at 3–5 dpi from ocular swabs by real-time RT-PCR targeting the HeV M gene. Virus titers in nasal swab samples were as high as 10^4.6 TCID₅₀/mL. The viral RNA was mainly distributed in tissues from respiratory and lymphoid systems at an early stage of infection and the presence of virus was confirmed by virus isolation. Pathological changes and immunohistochemical staining for viral antigen were consistent with the tissue distribution of the virus. This new finding indicates that pigs are susceptible to HeV infections and could potentially play a role as an intermediate host in transmission to humans.     

Transmission studies of Hendra virus (equine morbilli-virus) in fruit bats, horses and cats

Objective To determine the infectivity and transmissibility of Hendra virus (HeV). Design A disease transmission study using fruit bats, horses and cats. Procedure Eight grey-headed fruit bats (Pteropus poliocephalus) were inoculated and housed in contact with three uninfected bats and two uninfected horses. In a second exper iment, four horses were inoculated by subcutaneous injection and intranasal inoculation and housed in contact with three uninfected horses and six uninfected cats. In a third experi ment, 12 cats were inoculated and housed in contact with three uninfected horses. Two surviving horses were inoculated at the conclusion of the third experiment: the first orally and the second by nasal swabbing. All animals were necropsied and examined by gross and microscopic pathological methods, immunoperoxidase to detect viral antigen in formalin-fixed tissues, virus isolation was attempted on tissues and SNT and ELISA methods were used to detect HeV-specific antibody. Results Clinical disease was not observed in the fruit bats, although six of eight inoculated bats developed antibody against HeV, and two of six developed vascular lesions which contained viral antigen. The in-contact bats and horses did not seroconvert. Three of four horses that were inoculated devel oped acute disease, but in-contact horses and cats were not infected. In the third experiment, one of three in-contact horses contracted disease. At the time of necropsy, high titres of HeV were detected in the kidneys of six acutely infected horses, in the urine of four horses and the mouth of two, but not in the nasal cavities or tracheas. Conclusions Grey-headed fruit bats seroconvert and develop subclinical disease when inoculated with HeV. Horses can be infected by oronasal routes and can excrete HeV in urine and saliva. It is possible to transmit HeV from cats to horses. Transmission from P poliocephalus t o horses could not be proven and neither could transmission from horses to horses or horses to cats. Under the experimental conditions of the study the virus is not highly contagious.     

SARS,possible zoonosis in the area of conflict of pathogenic coronaviruses of animals

Severe acute respiratory syndrome (SARS) is an emerging disease, which was first recognized in Guangdong Province, China, in November 2002. In the meantime, SARS has been recognized in patients on all five continents. A novel coronavirus, which is not related to the hitherto known coronaviruses, has been proven to be associated with the disease. Our genomic analyses strongly suggest that the new SARS-coronavirus did not emerge through mutation or recombination and that it has probably been transmitted from a so far not identified animal species to humans. Therefore, it is most likely that SARS virus is a zoonotic agent. A broad body of knowledge originating from research in veterinary medicine indicates that development of vaccines against the SARS-coronavirus may be problematic. The potential danger of such vaccines should not be neglected during the process of vaccine development.     

Infectious disease surveillance of apparently healthy horses at a multi-day show using a novel nanoscale real-time PCR panel

In the United States, horses are used for a variety of purposes including recreation, exhibition, and racing. As farm, performance, and companion animals, horses are a unique species from a zoonotic disease risk perspective, and the risks of subclinical infections spreading among horses can pose challenges. Using a nanoscale real-time PCR platform, we investigated the prevalence of 14 enteric pathogens, 11 Escherichia coli genes, and 9 respiratory pathogens in fecal samples from 97 apparently healthy horses at a multi-day horse event. In addition, sugar flotation test was performed for fecal parasites. E. coli f17 was commonly detected, prevalent in 59% of horses, followed closely by Streptococcus equi subsp. zooepidemicus (55%). Additional pathogens recognized included betacoronavirus, Campylobacter jejuni, Cryptosporidium sp., E. coli O157, equine adenovirus 1, equine rhinitis B virus, and others. The use of PCR data may overestimate the true prevalence of these pathogens but provides a sensitive overview of common pathogens present in healthy horses. Our results prompt the continued need for practical biosecurity measures at horse shows, both to protect individuals interacting with these horses and to minimize transmission among horses.     

Anti-insect defensive behaviors in equines post-West Nile virus infection

West Nile virus (WNV), a zoonotic mosquito transmitted Flavivirus, has had significant health effects on horses in the United States, with over 23,000 United States equine cases since the disease was first recognized in 1999. Previous research has focused on how this disease progresses and affects equids days to weeks post infection. The purpose of this study was to evaluate if permanent equine behavioral changes had occurred in horses that had recovered from acute West Nile fever or encephalitis. Specifically, we examined if surviving this disease caused changes in the defensive behaviors of the animal against biting and stinging insects, presumably because of neurological sequelae that can result from the infection. Results from behavioral observations and neurologic reflex testing suggest that long-term survivors of WNV do not show a change in the frequency or types of behaviors used compared to uninfected horses, supporting the concept that lasting deficits from WNV usually resolve within the following 1–3 years post-infection. However, microhabitat and grouping behavior did have a significant impact on the frequency of defensive behaviors, with indoor locales and larger groups of horses showing less insect avoidance behaviors. These principles may play a more pivotal role in protecting equines from biting insects and disease than thought previously.     

Detection of the Severe Acute Respiratory Syndrome‐Related Coronavirus and Alphacoronavirus in the Bat Population of Taiwan

Bats have been demonstrated to be natural reservoirs of severe acute respiratory syndrome coronavirus (SARS CoV) and Middle East respiratory syndrome (MERS) CoV. Faecal samples from 248 individuals of 20 bat species were tested for partial RNA‐dependent RNA polymerase gene of CoV and 57 faecal samples from eight bat species were tested positive. The highest detection rate of 44% for Scotophilus kuhlii, followed by 30% for Rhinolophus monoceros. Significantly higher detection rates of coronaviral RNA were found in female bats and Scotophilus kuhlii roosting in palm trees. Phylogenetic analysis classified the positive samples into SARS‐related (SARSr) CoV, Scotophilus bat CoV 512 close to those from China and Philippines, and Miniopterus bat CoV 1A‐related lineages. Coronaviral RNA was also detected in bat guano from Scotophilus kuhlii and Myotis formosus flavus on the ground and had potential risk for human exposure. Diverse bat CoV with zoonotic potential could be introduced by migratory bats and maintained in the endemic bat population in Taiwan.     

Transmission ecology of rodent‐borne diseases: New frontiers

Rodents are recognized reservoir hosts for many human zoonotic pathogens. The current trends resulting from anthropocene defaunation suggest that in the future they, along with other small mammals, are likely to become the dominant mammals in almost all human‐modified environments. Recent intricate studies on bat‐borne emerging diseases have highlighted that many gaps exist in our understanding of the zoonotic transmission of rodent‐borne pathogens. This has emphasized the need for scientists interested in rodent‐borne diseases to integrate rodent ecology into their analysis of rodent‐borne pathogen transmission in order to identify in more detail the mechanisms of spillover and chains of transmission. Further studies are required to better understand the true impact of rodent abundance and the importance of pathogen sharing and circulation in multi‐host– multi‐pathogen communities. We also need to explore in more depth the roles of generalist and abundant species as the potential links between pathogen‐sharing, co‐infections and disease transmission.     

Implications of simian retroviruses for captive primate population management and the occupational safety of primate handlers.

Nonhuman primates can be naturally infected with a plethora of viruses with zoonotic potential, including retroviruses. These simian viruses present risks to both captive nonhuman primate populations and persons exposed to nonhuman primates. Simian retroviruses, including simian immunodeficiency virus, simian type D retrovirus, simian T-lymphotropic virus, and gibbon ape leukemia virus, have been shown to cause clinical disease in nonhuman primates. In contrast, simian foamy virus, a retrovirus that is highly prevalent in most nonhuman primates, has not been associated with clinical disease in naturally infected primates. Although it has been shown that human retrovirus infections with human T-lymphotropic virus and human immunodeficiency virus originated through multiple independent introductions of simian retroviruses into human populations that then spread globally, little is known about the frequency of such zoonotic events. In this article, exogenous simian retroviruses are reviewed as a concern for zoo and wildlife veterinarians, primate handlers, other persons in direct contact with nonhuman primates, and other nonhuman primates in a collection. The health implications for individual animals as well as managed populations in zoos and research institutions are discussed, the cross-species transmission and zoonotic disease potential of simian retroviruses are described, and suggestions for working safely with nonhuman primates are provided.