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Nipah virus causes outbreaks in humans with high case-fatality rates. In this study, we confirmed the presence of Nipah virus in Sri Lanka in Pteropus medius fruit bats, one of the known natural reservoir species. Sequences we generated were genetically related to Nipah virus strains from outbreaks in southern India.

The 2018 outbreak of Nipah virus in Kerala, India, highlights the need for global surveillance of henipaviruses in bats, which are the reservoir hosts for this and other viruses. Nipah virus, an emerging paramyxovirus in the genus Henipavirus, causes severe disease and stuttering chains of transmission in humans and is considered a potential pandemic threat. In May 2018, an outbreak of Nipah virus began in Kerala, > 1800 km from the sites of previous outbreaks in eastern India in 2001 and 2007. Twenty-three people were infected and 21 people died (16 deaths and 18 cases were laboratory confirmed). Initial surveillance focused on insectivorous bats (Megaderma spasma), whereas follow-up surveys within Kerala found evidence of Nipah virus in fruit bats (Pteropus medius). P. medius is the confirmed host in Bangladesh and is now a confirmed host in India. However, other bat species may also serve as reservoir hosts of henipaviruses. To inform surveillance of Nipah virus in bats, we reviewed and analyzed the published records of Nipah virus surveillance globally. We applied a trait-based machine learning approach to a subset of species that occur in Asia, Australia, and Oceana. In addition to seven species in Kerala that were previously identified as Nipah virus seropositive, we identified at least four bat species that, on the basis of trait similarity with known Nipah virus-seropositive species, have a relatively high likelihood of exposure to Nipah or Nipah-like viruses in India. These machine-learning approaches provide the first step in the sequence of studies required to assess the risk of Nipah virus spillover in India. Nipah virus surveillance not only within Kerala but also elsewhere in India would benefit from a research pipeline that included surveys of known and predicted reservoirs for serological evidence of past infection with Nipah virus (or cross reacting henipaviruses). Serosurveys should then be followed by longitudinal spatial and temporal studies to detect shedding and isolate virus from species with evidence of infection. Ecological studies will then be required to understand the dynamics governing prevalence and shedding in bats and the contacts that could pose a risk to public health.

Nipah virus, a zoonotic virus with a high mortality rate, threatens people from Indonesia because of its proximity to affected regions and the presence of bat reservoirs. Molecular screening of 64 Pteropus hypomelanus bats in Central Java detected 2 positive bats. Public health authorities should increase surveillance to help prevent human transmission.

Nipah virus (NiV) is a highly pathogenic zoonotic paramyxovirus that causes fatal encephalitis and respiratory disease in humans. There is currently no approved therapeutic for human use against NiV infection. Griffithsin (GRFT) is high-mannose oligosaccharide binding lectin that has shown in vivo broad-spectrum activity against viruses, including severe acute respiratory syndrome coronavirus, human immunodeficiency virus 1, hepatitis C virus, and Japanese encephalitis virus. In this study, we evaluated the in vitro antiviral activities of GRFT and its synthetic trimeric tandemer (3mG) against NiV and other viruses from 4 virus families. The 3mG had comparatively greater potency than GRFT against NiV due to its enhanced ability to block NiV glycoprotein-induced syncytia formation. Our initial in vivo prophylactic evaluation of an oxidation-resistant GRFT (Q-GRFT) showed significant protection against lethal NiV challenge in Syrian golden hamsters. Our results warrant further development of Q-GRFT and 3mG as potential NiV therapeutics.

Samples known or suspected to be infected with high-consequence viruses such as Ebola, Nipah, and Lassa must be handled under high biocontainment. Studies involving animal infections with these pathogens can generate tissues that require downstream analyses, including molecular assays and histopathology, which are more readily performed, or in some cases only feasible, at lower containment levels. Before removal from high containment for analyses at lower containment levels, specimens must undergo validated inactivation procedures. Here, we quantified viral load reduction in tissues infected with these pathogens following treatment with neutral-buffered formalin for 10 min, 1 h, 3 days, or 7 days, and MagMAX lysis/binding solution concentrate or TriPure isolation reagent for 1 or 10 min. To ensure accurate detection of any residual infectious virus, samples were purified through resins or centrifugal filters to reduce reagent cytotoxicity and maximize volume of testable material. We demonstrated effective inactivation (≥ 4 log 10 reduction) of all three pathogens and quantified log-reduction values over multiple timepoints. These findings provide validation data to support safe handling of infectious tissues for research, field studies, and outbreak response.

Hendra virus (HeV) and Nipah virus (NiV) are two highly pathogenic RNA viruses with zoonotic potential, which can cause severe diseases with high mortality rates (50–100%) in humans and animals. Given this context, these viruses are classified as Biosafety Level 4 (BSL-4) pathogens, thus limiting research studies. Despite the high case fatalities, there are currently no human vaccines available for either virus, owing in part to the limitations in research and hesitancy in funding. In the absence of widespread vaccination, diagnostic tests are crucial for the rapid identification of cases and disease surveillance. This review synthesizes current knowledge on the epidemiology, transmission dynamics, and pathogenesis of NiV and HeV to contextualize a detailed assessment of the available diagnostic tools. We examined molecular and serological assays, including RT-PCR, ELISA, and LAMP, highlighting sample sources, detection windows, and performance. Diagnostic considerations across human and animal hosts are discussed, with emphasis on outbreak applicability and field-readiness, given the need for diagnostic assays that are suitable for use in low-income areas. Further development of diagnostic assays, including isothermal amplification tests and other next-generation approaches, is recommended to fill the gap in rapid, point-of-care diagnostics.

Nipah virus (NiV) is an emerging high-fatality zoonotic threat lacking approved vaccines. Current virus-like particle (VLP) production methods rely on costly mammalian cell systems and non-scalable ultracentrifugation purification. We developed an alternative enveloped Nipah virus-like particle (NiVLP) expression system in Sf9 insect cells by co-expressing structural proteins F, G, and M using a single recombinant baculovirus. A novel multi-step chromatographic purification process was established using monolith convective media, integrating steric exclusion chromatography with sequential cation and anion exchange steps. Purified NiVLPs were characterized by nanoparticle tracking analysis and transmission electron microscopy, followed by immunogenicity study in Syrian golden hamsters. The optimized process yielded enveloped particles of approximately 100–120 nm that morphologically resemble native NiV virions. A single 25 µg NiVLP dose induced robust systemic anti-NiV G IgG responses within 14 days, demonstrating rapid immunogenicity suitable for outbreak response. However, neutralizing antibody titers against NiV remained limited compared to total IgG responses. This study establishes the first chromatography-based manufacturing platform for morphologically correct NiVLPs from insect cells. A deeper understanding of the immunity generated is needed to support their potential as a rapidly deployable vaccine platform against NiV.

Bats are natural reservoirs for a wide range of microorganisms, including many notable zoonotic pathogens. However, the composition of the infectome (i.e., the collection of viral, bacterial and eukaryotic microorganisms) within bat kidneys remains poorly understood. To address this gap, we performed meta-transcriptomic sequencing on kidney tissues from 142 bats, spanning ten species sampled at five locations in Yunnan province, China. This analysis identified 22 viral species, including 20 novel viruses, two of which represented newly discovered henipaviruses closely related to the highly pathogenic Hendra and Nipah viruses. These henipaviruses were found in the kidneys of bats inhabiting an orchard near villages, raising concerns about potential fruit contamination via bat urine and transmission risks to livestock or humans. Additionally, we identified a novel protozoan parasite, tentatively named Klossiella yunnanensis , along with two highly abundant bacterial species, one of which is a newly discovered species— Flavobacterium yunnanensis . These findings broaden our understanding of the bat kidney infectome, underscore critical zoonotic threats, and highlight the need for comprehensive, full-spectrum microbial analyses of previously understudied organs to better assess spillover risks from bat populations.

The Nipah Virus (NiV) is a zoonotic pathogen with the mortality rate of up to 75%, recurring in Asia over the past two decades. Due to increasing the risk of human transmission mediated by various intermediate hosts such as pigs and bats, it is necessary to produce an accurate and reliable point-of-care molecular detection method for NiV field diagnosis. In this study, we designed two pairs of primers targeting the conserved G and P genes and developed a point-of-care nucleic acid detection (POC-NAD) system by integrating one-step RT-PCR, lateral flow immunoassay, and microfluidic technologies. The POC-NAD system shows high specificity and sensitivity, with a Limit of Detection (LoD) of 199.1 copies/rxn. The primers aiming to the conserved sequences enables simultaneous detection of both NiV-M and NiV-B strains. Continuous evaluation of 21 simulated clinical samples demonstrated 100% concordance with RT-PCR results. Lateral flow-based visualization improves the display time and legibility of RT-PCR results. Additionally, microfluidic chips or chambers offer disposable reagent containers and consistent PCR amplification results across various field conditions. Therefore, the diagnostic tool is suitable for real-time nucleic acid testing and NiV surveillance in resource-limited field environments.

To better understand the potential risks of Nipah virus emergence in Cambodia by studying different components of the interface between humans and bats. From 2012 to 2016, we conducted a study at two sites in Kandal and Battambang provinces where fruit bats ( ) roost. We combined research on: bat ecology (reproductive phenology, population dynamics and diet); human practices and perceptions (ethnographic research and a knowledge, attitude and practice study); and Nipah virus circulation in bat and human populations (virus monitoring in bat urine and anti-Nipah-virus antibody detection in human serum). Our results confirmed circulation of Nipah virus in fruit bats (28 of 3930 urine samples positive by polymerase chain reaction testing). We identified clear potential routes for virus transmission to humans through local practices, including fruit consumed by bats and harvested by humans when Nipah virus is circulating, and palm juice production. Nevertheless, in the serological survey of 418 potentially exposed people, none of them were seropositive to Nipah virus. Differences in agricultural practices among the regions where Nipah virus has emerged may explain the situation in Cambodia and point to actions to limit the risks of virus transmission to humans. Human practices are key to understanding transmission risks associated with emerging infectious diseases. Social science disciplines such as anthropology need to be integrated in health programmes targeting emerging infectious diseases. As bats are hosts of major zoonotic pathogens, such integrated studies would likely also help to reduce the risk of emergence of other bat-borne diseases.