Objective:To evaluate immunogenicity, persistence of antibodies, survival after challenge, and anamnestic response following extralabel parenteral rabies vaccination across mammalian species compared with benchmarks from on-label canine vaccination. Methods:Mammals described in published studies of extralabel parenteral rabies vaccination. A structured literature review (1980 to 2025) identified studies reporting serologic or challenge outcomes after extralabel parenteral rabies vaccination. Extracted data included taxonomic family; vaccine type and schedule, timing of serum collection, antibody titers, survival after experimental challenge, and response to booster vaccination. Random-effects meta-analyses were conducted by family and overall. Pooled proportions were compared with the World Organisation for Animal Health benchmark of 88% survival and with canine benchmarks for primary seroconversion (88.1%), antibody levels 6 to 12 months after primary vaccination (72.9%), and booster response (96.8%). Results:32 articles describing 41 studies met inclusion criteria, representing 2,630 animals from 28 species in 17 families. After primary vaccination, 84.4 ± 4.1% achieved titers ≥ 0.5 IU/mL, with 14 of 16 families noninferior to the canine benchmark. At ≥ 6 months, 63.7 ± 10.6% maintained titers ≥ 0.5 IU/mL. Among 79 vaccinated animals challenged with rabies virus, 89.0 ± 9.8% survived. Booster vaccination produced adequate titers in 89.1 ± 8.0% of 275 animals, with reduced responses in the families Elephantidae and Ursidae. Conclusions:Modern-day rabies vaccines generate strong antibody responses across diverse mammalian families, with nondomestic canids responding similarly to domestic dogs. Clinical Relevance:These findings support recognizing extralabel vaccination as biologically protective and can inform public health and animal management policies.

Oral rabies vaccination (ORV) of dogs has been suggested as a complementary tool to injectable rabies vaccination, to access free-roaming dogs during mass dog vaccination campaigns. Stringent safety requirements have been developed and implemented for ORVs targeting wildlife and dogs. Part of the safety requirements involves conducting a risk assessment of the potential for human contact with the vaccine virus and the impact on human health. The former is predominantly determined by the bait distribution method and the latter by the safety profile of the vaccine construct. This risk assessment is focused on a commercially available ORV for dogs, SPBN GASGAS. Overall, it is concluded that although human contacts with the vaccine virus do occur infrequently, the risk of serious adverse events in humans is negligible due to the safety profile of the vaccine virus construct.

BACKGROUND:Rabies is one of the oldest known zoonoses. In Europe, because cases are rare and medical experience is limited, there is often uncertainty about the indications and modalities of pre- and post-exposure prophylaxis (PrEP, PEP). METHODS:This review is based on studies of the epidemiology, diagnosis, treatment, and prophylaxis of rabies that were published from January 2000 to October 2025 and retrieved by a selective search in PubMed, supplemented by data from the WHO, CDC, ECDC, RKI, and FLI and the current recommendations of the German Standing Committee on Vaccination (STIKO). RESULTS:The WHO reports that approximately 60 000 persons die of rabies each year around the world, mainly in southern Asia and Africa, and that dogs are the source of the infection in more than 99% of cases. Once the disease has become manifest, its lethality is nearly 100%. Terrestrial rabies is considered to have been largely eliminated in the member states of the European Union (EU) since 2008, although zoonotically relevant bat lyssaviruses are still being regularly documented. As a result, even in Germany, a low risk of infection remains through contact with bats and through importation of infected animals from endemic areas abroad. The use of modern cell-culture vaccines for PrEP yields protection rates of 96-100%, with rare severe side effects (< 1:10 000). PEP is highly effective: after its administration in more than 29 million cases per year around the world from 1980 to 2022, only 122 breakthrough infections were registered, mainly because of faulty administration of rabies immunoglobulin, inadequate wound cleansing, and a late start of PEP. The main preventive measures are PrEP, avoidance of exposure, rapid wound disinfection, early PEP, and counseling by specialists in travel medicine. CONCLUSION:Despite the favorable epidemiological situation, the possibility of rabies should be considered early on for anyone in Europe who has had an unclear animal exposure-especially to bats-or a potential rabies exposure abroad. When questions remain about the indication or modalities of PEP, expert advice should be obtained from an infectious disease unit or a rabies information center.

Efficacy studies are part of the regulatory requirements for obtaining marketing authorization for rabies virus vaccines intended for veterinary use. However, increasing emphasis on animal welfare and the application of the 3Rs principles (Replacement, Reduction, and Refinement) may prompt a critical reassessment of current standards for such studies. A substantial amount of data from numerous published experimental studies is available, which could be used to reconsider or refine existing specifications and protocols—particularly concerning observation periods post-infection (pi) and dose-finding studies. To support this reassessment, we analyzed data from 289 published experimental studies. These studies covered both susceptibility and efficacy investigations across 16 mesocarnivorous target species considered for parenteral or oral rabies vaccination. The species represented seven taxonomic families or orders: Canidae, Didelphidae, Felidae, Herpestidae, Mephitidae, Musteloidea, and Procyonidae. The dataset included 1,171 records encompassing a total of 8,381 animals that had been vaccinated with either rabies virus (RABV) or non-RABV lyssaviruses. Parametric distributions (log-normal, Weibull, and gamma) were fitted to incubation periods to examine group differences. Additionally, multiple linear and logistic regression analyses were performed to identify key variables influencing incubation duration, post-infection survival, and inter-species differences among reservoir hosts. The analyses revealed that viral strain, inoculation dose, and application site were consistently associated with incubation period length and mortality across most taxonomic groups. These variables significantly influenced incubation periods and survival rates post-infection. Notable differences were also observed between reservoir host species, highlighting the importance of species-specific factors in study design and interpretation. Our findings suggest that, in alignment with the 3Rs principle, existing animal models used in rabies vaccine efficacy research can be reconsidered and potentially refined. These refinements may not only enhance scientific validity but also contribute to ethical improvements in study design. Furthermore, the results may inform future risk assessments related to the importation of pets from rabies-endemic regions and support more humane, evidence-based approaches to vaccine evaluation.

In April 2024, following the annual International Committee on Taxonomy of Viruses (ICTV) ratification vote on newly proposed taxa, the phylum Negarnaviricota was expanded by 1 new order, 1 new family, 6 new subfamilies, 34 new genera and 270 new species. One class, two orders and six species were renamed. Seven families and 12 genera were moved; ten species were renamed and moved; and nine species were abolished. This article presents the updated taxonomy of Negarnaviricota as currently accepted by the ICTV, providing an essential annual update on the classification of members of this phylum that deepen understandings of their evolution, and supports critical public health measures for virus identification and tracking.

Ancient genome sequences provide invaluable insights into the origin and evolution of viral pathogens, offering a broader temporal perspective that extends well beyond the limited timespan of clinical data, which typically covers at most a few decades. Whereas ancient viral DNA is relatively frequently recovered, ancient viral RNA genomes are scarce due to the fragile nature of RNA molecules. In this study, we explored the feasibility of detecting ancient viral RNA in ethanol-preserved bat samples from a museum collection. We not only detected viral genome fragments but also recovered the coding-complete genome of a bandavirus (species Bandavirus zwieselense, family Phenuiviridae, order Hareavirales, class Bunyaviricetes) from a Common pipistrelle (Pipistrellus pipistrellus) bat collected in northern Germany in 1919. To investigate the modern distribution of Bandavirus zwieselense, we screened bat organs collected in Germany and the Netherlands via RT-qPCR, identifying modern counterparts in nine Common Pipistrelle (P. pipistrellus) bats (collected between 2010 and 2018), and one Serotine bat (Eptesicus serotinus, collected in 1999). The resulting genomic data enabled us to map phylogenetic relationships within this previously uncharacterized virus species (Bandavirus zwieselense) and estimate the timeframe for the most recent common ancestor. Additionally, we performed functional analysis of the S segment encoded nonstructural (NSs) protein of Bandavirus zwieselense in human HEK-293T cells, demonstrating its ability to block interferon induction, a characteristic also observed in the related human-pathogenic Severe fever with thrombocytopenia syndrome virus (SFTSV, species Bandavirus dabieense). This study demonstrates the feasibility of recovering and characterizing viral genomes from ethanol-preserved ancient RNA material, underscoring the significant potential of museum collections to contribute to the understanding of RNA virus evolution. ### Competing Interest Statement The authors have declared no competing interest.

Rabies, mankind's oldest known zoonotic disease, is caused by single-stranded, negative-sense RNA viruses of the Lyssavirus genus, family Rhabdoviridae of the Mononegavirales order. It is transmitted following the bites of infected mammals. The recognized etiological agents are classified into 16 recognized and 2 putative lyssavirus species, of which the rabies virus species (RABV) represent the prototype member of the genus. The reservoir for terrestrial rabies caused by RABV in its entire complexity is cryptic. Only a few abundant hosts in the orders Carnivora and Chiroptera are able to maintain viral RABV variants, also referred to as primary reservoir hosts, while the great majority of mammals, including humans, though susceptible to rabies, are considered spillover (dead-end) hosts. Distribution of terrestrial rabies encompasses all continents, with the exception of Antarctica and Australia, where it still poses a serious public health problem. Substantial progress in eliminating terrestrial rabies is limited to Western and Central Europe and parts of the Americas.

In 2021, a comprehensive dog demographic questionnaire combined with a KAP survey were conducted in the northern communal areas (NCAs) of Namibia with the aim of gaining a better understanding of dog populations, owner behaviour, and knowledge, attitudes and practices (KAP) relating to rabies. The survey of 3,726 households across the eight regions of the NCAs provided insights that will inform interventions in order to improve human rabies prevention and Namibia's dog rabies control strategy. The results showed a relatively low average human/dog ratio (HDR) of 5.4:1 indicating a surprisingly high dog population of at least 272,000 dogs in the NCAs, 93% of which appear to be owned but are free-roaming. Data analysis revealed opportunities but also highlighted needs for improvements in rabies surveillance and mass dog vaccinations. Although knowledge, attitude, and practice scores towards epidemiologic and clinical aspects, human rabies prevention, and dog rabies vaccination were deemed to be acceptable, the survey nevertheless revealed deficiencies in certain aspects in some of the population. Interestingly, data seemed to indicate relatively high dog bite incidences per 100,000 people, ranging between 262 and 1,369 and a certain number of unreported human rabies cases. Despite the very high number of dogs, only 50% of dog-owning households reported having vaccinated their dogs. In order to address these issues, the planning, announcement, and implementation of mass dog vaccination campaigns needs to be adapted to achieve adequate vaccination coverage. Another focus needs to be on rabies awareness and education if Namibia is to be significantly contributing to the global goal of "Zero by 30".

Background: Rabies in Turkey is maintained by dogs, but following a sustained spill-over, red fox mediated rabies had spread from the Aegean region to the central part of T & uuml;rkiye. During the past four years from 2019 to 2023 large scale efforts used oral rabies vaccination (ORV) to control rabies in red foxes. Here, we present the results of the largest ORV campaign on the Asian continent. Methods: ORV campaigns were carried out twice a year in spring and autumn with a targeted bait density of 20-23 baits/km(2). Monitoring of ORV campaigns included the GIS-based analyses of bait distribution, the assessment of bait uptake through biomarker detection and the determination of seroconversion (sero-positivity in ELISA) in the target species collected within the vaccination area. For determination of fox rabies incidence in vaccination areas as the main indicator of the performance of the ORV campaigns, epidemiological data was obtained from the national passive surveillance program. Results: Aerial bait distribution was highly accurate, with >99 % of baits being recorded from targeted zones, thus meeting the desired bait densities. Although the overall bait uptake (28.1 %; 95 %CI: 23.2-32.8) and seroprevalance (36.3 %; 95 %CI: 30.0-43.2) were low, rabies incidence drastically decreased in ORV areas and rabies was eliminated from western and central parts of Turkey, with no reported cases in foxes from ORV areas in 2022 and 2023. Conclusions: A large-scale ORV campaign against fox rabies using high quality vaccine baits and the GIS-aided and monitored bait distribution was able to control fox mediated rabies in the western and central parts of T & uuml;rkiye. Rabies control both in dogs and foxes should be expanded to cover also the eastern parts of T & uuml;rkiye, to become eventually rabies free.

Vaccination of the reservoir species is a key component in the global fight against rabies. For wildlife reservoir species and hard to reach spillover species (e. g. ruminant farm animals), oral vaccination is the only solution. In search for a novel potent and safe oral rabies vaccine, we generated a recombinant vector virus based on lentogenic Newcastle disease virus (NDV) strain Clone 30 that expresses the glycoprotein G of rabies virus (RABV) vaccine strain SAD L16 (rNDV_GRABV). Transgene expression and virus replication was verified in avian and mammalian cells. To test immunogenicity and viral shedding, in a proof-of-concept study six goats and foxes, representing herbivore and carnivore species susceptible to rabies, each received a single dose of rNDV_GRABV (108.5 TCID50/animal) by direct oral application. For comparison, three animals received the similar dose of the empty viral vector (rNDV). All animals remained clinically inconspicuous during the trial. Viral RNA could be isolated from oral and nasal swabs until four (goats) or seven days (foxes) post vaccination, while infectious NDV could not be re-isolated. After four weeks, three out of six rNDV_GRABV vaccinated foxes developed RABV binding and virus neutralizing antibodies. Five out of six rNDV_GRABV vaccinated goats displayed RABV G specific antibodies either detected by ELISA or RFFIT. Additionally, NDV and RABV specific T cell activity was demonstrated in some of the vaccinated animals by detecting antigen specific interferon γ secretion in lymphocytes isolated from pharyngeal lymph nodes. In conclusion, the NDV vectored rabies vaccine rNDV_GRABV was safe and immunogenic after a single oral application in goats and foxes, and highlight the potential of NDV as vector for oral vaccines in mammals.

(1) Background: The oral vaccination of free-roaming dogs against rabies has been developed as a promising complementary tool for mass dog vaccination. However, no oral rabies vaccine has provided efficacy data in dogs according to international standards. (2) Methods: To test the immunogenicity and efficacy of the third-generation oral rabies virus vaccine strain, SPBN GASGAS, in domestic dogs, dogs were offered an egg-flavoured bait containing 3.0 mL of the vaccine (107.5 FFU/mL) or a placebo egg-flavoured bait. Subsequently, these 25 vaccinated and 10 control animals were challenged approximately 6 months later with a dog rabies virus isolate. Blood samples were collected at different time points postvaccination and examined by ELISA and RFFIT. (3) Results: All but 1 of the 25 vaccinated dogs survived the challenge infection; meanwhile, all 10 control dogs succumbed to rabies. The serology results showed that all 25 vaccinated dogs seroconverted in ELISA (>40% PB); meanwhile, only 13 of the 25 vaccinated dogs tested seropositive ≥ 0.5 IU/mL) in RFFIT. (4) Conclusions: The SPBN GASGAS rabies virus vaccine meets the efficacy requirements for live oral rabies vaccines as laid down by the European Pharmacopoeia and the WOAH Terrestrial Manual. SPBN GASGAS already fulfilled the safety requirements for oral rabies vaccines targeted at dogs. Hence, the egg-flavoured bait containing SPBN GASGAS is the first oral vaccine bait that complies with WOAH recommendations for the intended use of oral vaccination of free-roaming dogs against rabies.

In a long-term immunogenicity study (1100 days post vaccination) in local Thai dogs the immune response of the oral rabies vaccine SPBN GASGAS was compared to those elicited by a commercial inactivated vaccine using immunobridging. Based on the detection of rabies virus binding (rVBA) and rabies virus neutralizing antibodies (rVNA) as measured by ELISA and Rapid Fluorescent Focus Inhibition Test (RFFIT) the long-term immune response in dogs vaccinated orally with the SPBNA GASGAS strain of rabies vaccine in a bait was non-inferior to a conventional inactivated rabies vaccine. The outcome of this study supports extending the originally claimed duration of immunity (DOI) of SPBN GASGAS after oral vaccination for dogs from 6 to 30 months.

Rabies is endemic in Namibia and is present both in wildlife carnivores and domestic free-roaming dogs. The disease thus represents a challenge for public human and veterinary disease control. Namibia has implemented a national strategic plan to control rabies and the country’s activities are supported by international organizations. To this end, rabies diagnosis at the Central Veterinary Laboratory (CVL) was improved in the frame of a World Organization for Animal Health (WOAH) laboratory twinning program: from practical sampling techniques and the use of lateral flow devices to a novel universal and discriminatory quantitative real-time Reverse transcription polymerase chain reaction (RT-qPCR), which easily identify dog-associated rabies viruses. The procedures applied and the results can be used as a template to improve rabies laboratory diagnosis.

Dog-mediated rabies is responsible for tens of thousands of human deaths annually, and in resource-constrained settings, vaccinating dogs to control the disease at source remains challenging for various reasons. Currently, rabies elimination efforts rely on mass dog vaccination by the parenteral route. While oral rabies vaccination (ORV) of dogs is primarily considered a tool to increase herd immunity, particularly by targeting free-roaming and stray dogs, here, we are showcasing an ORV-only approach as an emergency response model. Using a third-generation vaccine and a standardized egg-flavored bait, we assessed the effectiveness and vaccination under field conditions in the Zambezi region of Namibia. During this trial, with four teams and within four working days, 3097 dogs were offered a bait, of which 88,0% were considered vaccinated. Teams managed to vaccinate, on average, over 20 dogs/h, despite using a door-to-door vaccination approach. The favorable results both in terms of bait acceptance and successful vaccination as well as field applicability and effectiveness further support the great potential of ORV in dog rabies control programmes.

Background The high susceptibility of carnivores to Suid Alphaherpesvirus 1 [SuAHV1, synonymous pseudorabies virus (PrV)], renders them inadvertent sentinels for the possible occurrence of Aujeszky’s disease (AD) in domestic and wild swine populations. The aim of this study was to epidemiologically analyse the occurrence of PrV infections in domestic and wild animals in Germany during the last three decades and to genetically characterise the causative PrV isolates. Methods PrV in dogs was detected using standard virological techniques including conventional and real time PCR, virus isolation or by immunohistochemistry. Available PrV isolates were characterized by partial sequencing of the open gC reading frame and the genetic traits were compared with those of archived PrV isolates from carnivores and domestic pigs from Germany before the elimination of AD in the domestic pig population. Results During 1995 and 2022, a total of 38 cases of AD in carnivores, e.g. dogs and red foxes, were laboratory confirmed. Sequencing and subsequent phylogenetic analysis of PrV isolates established a strong connection between AD cases in carnivores and the occurrence of PrV infections in European wild boars in the end phase of and after elimination of AD from the domestic pig population. While PrV infections occur at low numbers but regularly in hunting dogs, interestingly, PrV was not observed in grey wolves in Germany. In none of 682 dead-found grey wolves and wolf-dog hybrids tested from Germany during 2006–2022 could PrV infection be detected by molecular means. Conclusions Although PrV has been eliminated from domestic pigs, spillover infections in domestic and wild carnivores should always be expected given the endemic presence of PrV in wild pig populations. Since detection of PrV DNA and virus in carnivores is sporadic even in areas with high seroprevalence of PrV in wild pigs, it may not reflect the full diversity of PrV.

EDITORIAL article Front. Vet. Sci., 18 July 2023Sec. Veterinary Epidemiology and Economics Volume 10 - 2023 | https://doi.org/10.3389/fvets.2023.1220327

Despite being vaccine preventable, dog-mediated rabies continues unabated in low-resourced countries in Africa and Asia. For interventions into dog rabies control, an estimate of the dog population is a prerequisite. Here we used a High-Resolution Settlement Layer (HRSL) with an unprecedented resolution of 30m grid length that is Open Source for dog populations estimates and studies on vaccination coverages, with the Oshana region of Namibia as an example. Our analyses show that the average dog density per km 2 is 8.15 but ranges between 0 and 40 per constituency, with individual densities being as high as 551. Spatial analyses for different settings of static vaccination points indicate that the previously used vaccination points during the pilot phase and cattle crush-pens are insufficient for reaching a 70% vaccination level in the Oshana region. Based on cost calculations, between US$5.29 and US$7.77 are needed to parenterally vaccinate dogs in this region, suggesting that oral rabies vaccination may be a cost-effective supplement or even replacement. The high-resolution spatial analyses are exemplified for rabies, but any other One Health intervention, particularly for Neglected tropical diseases in highly heterogenous and remote areas could use our approach as a template. Author Summary Here, we used high-resolution geospatial data for demonstrating and validating its utility to assist veterinary authorities in their fight against dog-mediated rabies on the example of the Oshana region in Namibia. With such detailed data it is possible estimate the dog population and to analyse and optimize vaccination strategies in dog rabies endemic areas. Such analyses are exemplified for rabies, but any other One Health intervention, particularly for Neglected tropical diseases in highly heterogenous and remote areas could use our approach as a template.

Licensed rabies virus vaccines based on whole inactivated virus are effective in humans. However, there is a lack of detailed investigations of the elicited immune response, and whether responses can be improved using novel vaccine platforms. Here we show that two doses of a lipid nanoparticle-formulated unmodified mRNA vaccine encoding the rabies virus glycoprotein (RABV-G) induces higher levels of RABV-G specific plasmablasts and T cells in blood, and plasma cells in the bone marrow compared to two doses of Rabipur in non-human primates. The mRNA vaccine also generates higher RABV-G binding and neutralizing antibody titers than Rabipur, while the degree of somatic hypermutation and clonal diversity of the response are similar for the two vaccines. The higher overall antibody titers induced by the mRNA vaccine translates into improved cross-neutralization of related lyssavirus strains, suggesting that this platform has potential for the development of a broadly protective vaccine against these viruses.