Henipavirus

Henipavirus is a genus of the family Paramyxoviridae, order Mononegavirales containing three established species: Hendra virus, Nipah virus and Cedar Virus. The henipaviruses are naturally harboured by Pteropid fruit bats (flying foxes) and some microbat species.^[1] Henipavirus is characterised by a large genome, a wide host range, and their recent emergence as zoonotic pathogens capable of causing illness and death in domestic animals and humans.^[2]

In 2009, RNA sequences of three novel viruses in phylogenetic relationship to known Henipaviruses were detected in Eidolon helvum (the African Straw-coloured fruit bat) in Ghana. The finding of these novel putative Henipaviruses outside Australia and Asia indicates that the region of potential endemicity of Henipaviruses extends to Africa.^[3]

Virus structure

Structure of henipaviruses

Henipaviruses are pleomorphic (variably shaped), ranging in size from 40 to 600 nm in diameter.^[4] They possess a lipid membrane overlying a shell of viral matrix protein. At the core is a single helical strand of genomic RNA tightly bound to N (nucleocapsid) protein and associated with the L (large) and P (phosphoprotein) proteins which provide RNA polymerase activity during replication.

Embedded within the lipid membrane are spikes of F (fusion) protein trimers and G (attachment) protein tetramers. The function of the G protein is to attach the virus to the surface of a host cell via EFNB2, a highly conserved protein present in many mammals.^[5][6] The F protein fuses the viral membrane with the host cell membrane, releasing the virion contents into the cell. It also causes infected cells to fuse with neighbouring cells to form large, multinucleated syncytia.

Genome structure

As with all viruses in the Mononegavirales order, the Hendra virus and Nipah virus genomes are non-segmented, single-stranded negative-sense RNA. Both genomes are 18.2 kb in size and contain six genes corresponding to six structural proteins.^[7]

In common with other members of the Paramyxovirinae subfamily, the number of nucleotides in the henipavirus genome is a multiple of six, consistent with what is known as the 'rule of six'.^[8] Deviation from the rule of six, through mutation or incomplete genome synthesis, leads to inefficient viral replication, probably due to structural constraints imposed by the binding between the RNA and the N protein.

Henipaviruses employ an unusual process called RNA editing to generate multiple proteins from a single gene. The specific process in henipaviruses involves the insertion of extra guanosine residues into the P gene mRNA prior to translation. The number of residues added determines whether the P, V or W proteins are synthesised. The functions of the V and W proteins are unknown, but they may be involved in disrupting host antiviral mechanisms.

Hendra virus

Emergence

Hendra virus (originally Equine morbillivirus) was discovered in September 1994 when it caused the deaths of thirteen horses, and a trainer at a training complex in Hendra, a suburb of Brisbane in Queensland, Australia.^[9]

The index case, a mare, was housed with 19 other horses after falling ill, and died two days later. Subsequently, all of the horses became ill, with 13 dying. The remaining 6 animals were subsequently euthanised as a way of preventing relapsing infection and possible further transmission.^[10] The trainer, Victory ('Vic') Rail, and a stable hand were involved in nursing the index case, and both fell ill with an influenza-like illness within one week of the first horse’s death. The stable hand recovered while Mr Rail died of respiratory and renal failure. The source of the virus was most likely frothy nasal discharge from the index case.

A second outbreak occurred in August 1994 (chronologically preceding the first outbreak) in Mackay 1,000 km north of Brisbane resulting in the deaths of two horses and their owner.^[11] The owner, Mark Preston, assisted in necropsies of the horses and within three weeks was admitted to hospital suffering from meningitis. Mr Preston recovered, but 14 months later developed neurologic signs and died. This outbreak was diagnosed retrospectively by the presence of Hendra virus in the brain of the patient.^[12]

A survey of wildlife in the outbreak areas was conducted, and identified pteropid fruit bats as the most likely source of Hendra virus, with a seroprevalence of 47%. All of the other 46 species sampled were negative. Virus isolations from the reproductive tract and urine of wild bats indicated that transmission to horses may have occurred via exposure to bat urine or birthing fluids.^[13] However, the only attempt at experimental infection reported in the literature, conducted at CSIRO Geelong, did not result in infection of a horse from infected flying foxes. This study looked at potential infection between bats, horses and cats, in various combinations. The only species that was able to infect horses was the cat (Felix spp.).^[14]

Australian outbreaks

Hendra

Mackay

Townsville

Bowen

Hervey Bay

Chinchilla

Rockhampton

Ingham

Cairns

Outbreaks in Queensland

Ballina 

Macksville

Kempsey

Outbreaks in New South Wales

As of September 2012, a total of thirty-nine outbreaks of Hendra virus have occurred in Australia, all involving infection of horses. As a result of these events, seventy-six horses have died or been euthanised. A further four died or were euthanised as a result of possible hendra infection. Case fatality rate in humans is 60% and in horses 75%.^[15]

Four of these outbreaks have spread to humans as a result of direct contact with infected horses. On 26 July 2011 a dog living on the Mt Alford property was reported to have HeV antibodies, the first time an animal other than a flying fox, horse, or human has tested positive outside an experimental situation.^[16]

These events have all been on the east coast of Australia, with the most northern event at Cairns, Queensland and the event furthest south at Kempsey, NSW. Until the event at Chinchilla, Queensland in July 2011, all outbreak sites had been within the distribution of at least two of the four mainland flying-foxes (fruit bats); Little red flying-fox, (Pteropus scapulatus), black flying-fox, (Pteropus alecto), grey-headed flying-fox, (Pteropus poliocephalus) and spectacled flying-fox, (Pteropus conspicillatus). Chinchilla is considered to be only within the range of little red flying-fox and is west of the Great Dividing Range. This is the furthest west the infection has ever been identified in horses.

The timing of incidents indicates a seasonal pattern of outbreaks, possibly related to the breeding cycle of the little red flying-foxes. These species typically give birth between April and May.^[17][18] There is no evidence of transmission to humans directly from bats, and, as such it appears that human infection only occurs via an intermediate host, a horse.^[19]

Timing of outbreaks also appears more likely during the cooler months when it is possible the temperature and humidity are more favourable to the longer term survival of the virus in the environment.^[20]

Events of June – August 2011

In the years 1994–2010, fourteen events were recorded. Between 20 June 2011 and 28 August 2011, a further seventeen events were identified, during which twenty-one horses died.

It's not clear why there has been a sudden increase in the number of spillover events between June and August 2011. Typically HeV spillover events are more common between May and October. This time is sometimes called "Hendra Season".,^[85] which is a time when there are large numbers of fruit bats of all species congregated in SE Queensland's valuable winter foraging habitat. The weather (warm and humid) is favourable to the survival of henipavirus in the environment.^[86]

It is possible flooding in SE Queensland and Northern NSW in December 2010 and January 2011 may have had an impact on the health of the fruit bats. Urine sampling in flying-fox camps indicate that a larger proportion of flying-foxes than usual are shedding live virus. Biosecurity Queensland's ongoing surveillance usually shows 7% of the animals are shedding live virus. In June and July nearly 30% animals have been reported to be shedding live virus.^[87] Present advice is that these events are not being driven by any mutation in HeV itself.^[88]

Other suggestions include that an increase in testing has led to an increase in detection. As the actual mode of transmission between bats and horses has not been determined, it is not clear what, if any, factors can increase the chance of infection in horses.^[89]

Following the confirmation of a dog with HeV antibodies, on 27 July 2011, the Queensland and NSW governments will boost research funding into the Hendra virus by $6 million to be spent by 2014/2015. This money will be used for research into ecological drivers of infection in the bats and the mechanism of virus transmission between bats and other species.^[90][91] A further 6 million dollars was allocated by the federal government with the funds being split, half for human health investigations and half for animal health and biodiversity research.^[92]

Prevention, detection and treatment

Three main approaches are currently followed to reduce the risk to humans.^[93]

• Vaccine for horses. In November 2012, a vaccine became available for horses. The vaccine is to be used in horses only, since, according to CSIRO veterinary pathologist Dr Deborah Middleton, breaking the transmission cycle from flying foxes to horses prevents it from passing to humans, as well as, "a vaccine for people would take many more years."^[94][95]

The vaccine is a subunit vaccine that neutralises Hendra virus and is composed of a soluble version of the G surface antigen on Hendra virus and has been successful in ferret models.^[96][97][98]

• Stall-side test to assist in diagnosing the disease in horses rapidly. Although the research on the Hendra virus detection is undergoing, a promising result has found using antibody-conjugated magnetic particles and quantum dots.^[99][100]
• Post-exposure treatment for humans.

Nipah virus and Hendra virus are closely related paramyxoviruses that emerged from bats during the 1990s to cause deadly outbreaks in humans and domesticated animals. National Institute of Allergy and Infectious Diseases (NIAID)-supported investigators developed vaccines for Nipah and Hendra virus based on the soluble G-glycoproteins of the viruses formulated with adjuvants. Both vaccines have been shown to induce strong neutralizing antibodies in different laboratory animals.

Pathology

Flying foxes experimentally infected with the Hendra virus develop a viraemia then excrete the virus in their urine, faeces and saliva for approximately one week. Although they excrete live virus during this time there is no other indication of an illness.^[101] Symptoms of Hendra virus infection of humans may be respiratory, including hemorrhage and edema of the lungs, or encephalitic, resulting in meningitis. In horses, infection usually causes pulmonary oedema, congestion and / or neurological signs.^[102]

Ephrin B2 has been identified as the main receptor for the henipaviruses.^[103]

Hendra virus has been classified as a Bio-safety Level 4 Hot Agent.

Nipah virus

Emergence

Pteropus vampyrus (Large flying fox), one of the natural reservoirs of Nipah virus

Nipah virus was identified in April 1999, when it caused an outbreak of neurological and respiratory disease on pig farms in peninsular Malaysia, resulting in 257 human cases, including 105 human deaths and the culling of one million pigs.^[11][104] In Singapore, 11 cases, including one death, occurred in abattoir workers exposed to pigs imported from the affected Malaysian farms. The Nipah virus has been classified by the Centers for Disease Control and Prevention as a Category C agent.^[105] The name "Nipah" refers to the place, Kampung Baru Sungai Nipah in Negeri Sembilan State, Malaysia, the source of the human case from which Nipah virus was first isolated.^[106]

The outbreak was originally mistaken for Japanese encephalitis (JE), however, physicians in the area noted that persons who had been vaccinated against JE were not protected, and the number of cases among adults was unusual^[107] Despite the fact that these observations were recorded in the first month of the outbreak, the Ministry of Health failed to react accordingly, and instead launched a nationwide campaign to educate people on the dangers of JE and its vector, Culex mosquitoes.

Symptoms of infection from the Malaysian outbreak were primarily encephalitic in humans and respiratory in pigs. Later outbreaks have caused respiratory illness in humans, increasing the likelihood of human-to-human transmission and indicating the existence of more dangerous strains of the virus.

Based on seroprevalence data and virus isolations, the primary reservoir for Nipah virus was identified as Pteropid fruit bats, including Pteropus vampyrus (Large Flying Fox), and Pteropus hypomelanus (Small Flying-fox), both of which occur in Malaysia.

The transmission of Nipah virus from flying foxes to pigs is thought to be due to an increasing overlap between bat habitats and piggeries in peninsular Malaysia. At the index farm, fruit orchards were in close proximity to the piggery, allowing the spillage of urine, faeces and partially eaten fruit onto the pigs.^[108] Retrospective studies demonstrate that viral spillover into pigs may have been occurring in Malaysia since 1996 without detection.^[11] During 1998, viral spread was aided by the transfer of infected pigs to other farms, where new outbreaks occurred.

Outbreaks

Locations of henipavirus outbreaks (red stars–Hendra virus; blue stars–Nipah virus) and distribution of henipavirus flying fox reservoirs (red shading–Hendra virus ; blue shading–Nipah virus)

Eight more outbreaks of Nipah virus have occurred since 1998, all within Bangladesh and neighbouring parts of India. The outbreak sites lie within the range of Pteropus species (Pteropus giganteus). As with Hendra virus, the timing of the outbreaks indicates a seasonal effect. Cases occurring in Bangladesh during the winters of 2001, 2003, and 2004, were determined to have been caused by the Nipah virus.^[109] In February 2011, a Nipah outbreak began at Hatibandha Upazila in the Lalmonirhat District of northern Bangladesh. To date (7 February 2011), there have been 24 cases and 17 deaths in this outbreak.^[110]

• 2001 January 31–23 February, Siliguri, India: 66 cases with a 74% mortality rate.^[111] 75% of patients were either hospital staff or had visited one of the other patients in hospital, indicating person-to-person transmission.
• 2001 April – May, Meherpur District, Bangladesh: 13 cases with nine fatalities (69% mortality).^[112]
• 2003 January, Naogaon District, Bangladesh: 12 cases with eight fatalities (67% mortality).^[112]
• 2004 January – February, Manikganj and Rajbari districts, Bangladesh: 42 cases with 14 fatalities (33% mortality).
• 2004 19 February – 16 April, Faridpur District, Bangladesh: 36 cases with 27 fatalities (75% mortality). 92% of cases involved close contact with at least one other person infected with Nipah virus. Two cases involved a single short exposure to an ill patient, including a rickshaw driver who transported a patient to hospital. In addition, at least six cases involved acute respiratory distress syndrome which has not been reported previously for Nipah virus illness in humans. This symptom is likely to have assisted human-to-human transmission through large droplet dispersal. ^{[citation needed]}
• 2005 January, Tangail District, Bangladesh: 12 cases with 11 fatalities (92% mortality). The virus was probably contracted from drinking date palm juice contaminated by fruit bat droppings or saliva.^[113]
• 2007 February – May, Nadia District, India: up to 50 suspected cases with 3–5 fatalities. The outbreak site borders the Bangladesh district of Kushtia where eight cases of Nipah virus encephalitis with five fatalities occurred during March and April 2007. This was preceded by an outbreak in Thakurgaon during January and February affecting seven people with three deaths.^[114] All three outbreaks showed evidence of person-to-person transmission.
• 2008 February – March, Manikganj and Rajbari districts, Bangladesh: Nine cases with eight fatalities.^[115]

• 2010 January, Bhanga subdistrict, Faridpur, Bangladesh: Eight cases with seven fatalities. During March, one physician of Faridpur Medical College Hospital caring for confirmed Nipah cases died^[116]

• 2011 February: An outbreak of Nipah Virus has occurred at Hatibandha, Lalmonirhat, Bangladesh. The deaths of 21 schoolchildren due to Nipah virus infection were recorded on 4 February 2011. IEDCR has confirmed the infection is due to this virus.^[117] Local schools were closed for one week to prevent the spread of the virus. People were also requested to avoid consumption of uncooked fruits and fruit products. Such foods, contaminated with urine or saliva from infected fruit bats, were the most likely source of this outbreak.^[118]

Nipah virus has been isolated from Lyle's flying fox (Pteropus lylei) in Cambodia^[119] and viral RNA found in urine and saliva from P. lylei and Horsfield's roundleaf bat (Hipposideros larvatus) in Thailand.^[120] Infective virus has also been isolated from environmental samples of bat urine and partially eaten fruit in Malaysia.^[121] Antibodies to henipaviruses have also been found in fruit bats in Madagascar (Pteropus rufus, Eidolon dupreanum)^[122] and Ghana (Eidolon helvum)^[123] indicating a wide geographic distribution of the viruses. No infection of humans or other species have been observed in Cambodia, Thailand or Africa thus far.

Pathology

In humans, the infection presents as fever, headache and drowsiness. Cough, abdominal pain, nausea, vomiting, weakness, problems with swallowing and blurred vision are relatively common. About a quarter of the patients have seizures and about 60% become comatose and might need mechanical ventilation. In patients with severe disease, their conscious state may deteriorate and they may develop severe hypertension, fast heart rate, and very high temperature.

Nipah virus is also known to cause relapse encephalitis. In the initial Malaysian outbreak, a patient presented with relapse encephalitis some 53 months after his initial infection. There is no definitive treatment for Nipah encephalitis, apart from supportive measures, such as mechanical ventilation and prevention of secondary infection. Ribavirin, an antiviral drug, was tested in the Malaysian outbreak, and the results were encouraging, though further studies are still needed.

While no vaccine currently exists, a recent (2012) study of a trial vaccine developed using the outer proteins of Hendra virus was shown to induce protection against Nipah in African Green Monkeys.^[124]

In animals, especially in pigs, the virus causes a porcine respiratory and neurologic syndrome, locally known as "barking pig syndrome" or "one mile cough."

Ephrin B2 has been identified as the main receptor for the henipaviruses.^[103]

In popular culture

Nipah virus was the inspiration for the MEV-1 virus in the film Contagion.^[125]

In the pilot episode of The Blacklist, Ranko Zamani (Jamie Jackson) has contracted the Nipah virus and wants to launch a chemical weapons attack in Washington D.C. before he dies.

Cedar virus

Emergence

Cedar Virus (CedPV) was first identified in pteropid urine during work on Hendra virus undertaken in Queensland in 2009.^[126]

Although the virus is reported to be very similar to both Hendra and Nipah, it does not cause illness in laboratory animals usually susceptible to paramyxoviruses. Animals were able to mount an effective response and create effective antibodies.^[126]

The scientists who identified the virus report:

Hendra and Nipah viruses are 2 highly pathogenic paramyxoviruses that have emerged from bats within the last two decades. Both are capable of causing fatal disease in both humans and many mammal species. Serological and molecular evidence for henipa-like viruses have been reported from numerous locations including Asia and Africa, however, until now no successful isolation of these viruses have been reported. This paper reports the isolation of a novel paramyxovirus, named Cedar virus, from fruit bats in Australia. Full genome sequencing of this virus suggests a close relationship with the henipaviruses. Antibodies to Cedar virus were shown to cross react with, but not cross neutralize Hendra or Nipah virus. Despite this close relationship, when Cedar virus was tested in experimental challenge models in ferrets and guinea pigs, we identified virus replication and generation of neutralizing antibodies, but no clinical disease was observed. As such, this virus provides a useful reference for future reverse genetics experiments to determine the molecular basis of the pathogenicity of the henipaviruses.^[126]

Causes of emergence

The emergence of henipaviruses parallels the emergence of other zoonotic viruses in recent decades. SARS coronavirus, Australian bat lyssavirus, Menangle virus and probably Ebola virus and Marburg virus are also harbored by bats and are capable of infecting a variety of other species. The emergence of each of these viruses has been linked to an increase in contact between bats and humans, sometimes involving an intermediate domestic animal host. The increased contact is driven both by human encroachment into the bats’ territory (in the case of Nipah, specifically pigpens in said territory) and by movement of bats towards human populations due to changes in food distribution and loss of habitat.

There is evidence that habitat loss for flying-foxes, both in South Asia and Australia (particularly along the east coast) as well as encroachment of human dwellings and agriculture into the remaining habitats, is creating greater overlap of human and flying fox distributions.

See also

• Animal viruses
• Paramyxovirus

References

External links

• Biosecurity Queensland Hendra virus
• Hendra virus
• ViralZone: Henipavirus
• Hendra virus factsheet – CSIRO
• Nipah virus – CSIRO
• Henipavirus – Henipavirus Ecology Research Group (HERG) INFO
• The science and mystery of hendra virus – by Renee du Preez (ABC Rural) – Australian Broadcasting Corporation.
• Enserink M (February 2009). "Virus's Achilles' Heel Revealed". Science Now. AAAS. 
• Virus Pathogen Database and Analysis Resource (ViPR): Paramyxoviridae