Dog Rabies Virus Research Articles

Genomic Characterization of a Dog-Mediated Rabies Outbreak in El Pedregal, Arequipa, Peru.

Rabies, a re-emerging zoonosis with the highest known human case fatality rate, has been largely absent from Peru, except for endemic circulation in the Puno region on the Bolivian border and re-emergence in Arequipa City in 2015, where it has persisted. In 2021, an outbreak occurred in the rapidly expanding city of El Pedregal near Arequipa, followed by more cases in 2022 after nearly a year of epidemiological silence. While currently under control, questions persist regarding the origin of the El Pedregal outbreak and implications for maintaining rabies control in Peru. We sequenced 25 dog rabies virus (RABV) genomes from the El Pedregal outbreak (n=11) and Arequipa City (n=14) from 2021-2023 using Nanopore sequencing in Peru. Historical genomes from Puno (n=4, 2010-2012) and Arequipa (n=5, 2015-2019), were sequenced using an Illumina approach in the UK. In total, 34 RABV genomes were analyzed, including archived and newly obtained samples. The genomes were analyzed phylogenetically to understand the outbreak's context and origins. Phylogenomic analysis identified two genetic clusters in El Pedregal: 2021 cases stemmed from a single introduction unrelated to Arequipa cases, while the 2022 sequence suggested a new introduction from Arequipa rather than persistence. In relation to canine RABV diversity in Latin America, all new sequences belonged to a new minor clade, Cosmopolitan Am5, sharing relatives from Bolivia, Argentina, and Brazil. Genomic insights into the El Pedregal outbreak revealed multiple introductions over a 2-year window. Eco-epidemiological conditions, including migratory worker patterns, suggest human-mediated movement drove introductions. Despite outbreak containment, El Pedregal remains at risk of dog-mediated rabies due to ongoing circulation in Arequipa, Puno, and Bolivia. Human-mediated movement of dogs presents a major risk for rabies re-emergence in Peru, jeopardizing regional dog-mediated rabies control. Additional sequence data is needed for comprehensive phylogenetic analyses.

Factors Involved in the Immunological Protection against Rabies Virus in Dogs in Spain.

Rabies, a viral disease spread by infected animal bites that causes encephalitis in humans and other mammals, is a neglected infectious disease present on all continents except Antarctica. Spain has been free of terrestrial rabies since 1978. However, due to its geographical situation, it represents a bridge for imported cases from an endemic continent such as Africa to Europe. Rabies vaccination in dogs is an essential preventive tool against this zoonosis. The aim of this study was to determine the state of the immune response against rabies virus in dogs in Spain and to demonstrate whether several factors that have been previously related to the influence of the seroprevalence of this species are involved here. The seroconversion level of this zoonotic virus was assessed in a total of 1060 animals. Indirect ELISA was used to obtain data for statistical analysis to evaluate the studied variables. Working under the concept of One Health, this study provides relevant information to be taken into consideration not only to prevent re-emergence in countries free of this disease but also for prevention and control in endemic countries.

A case of rabies outbreak in a bull-calf from Nigeria

This paper describes a rabies outbreak in a bull-calf which led to euthanasia following manifestation of clinical signs of the disease. Infection was confirmed using a rapid immunochromatographic test of the homogenates from brain tissues (the brain stem, hippocampus and cerebellum) sample. Exposure to rabies virus (RABV) had resulted due to an attack by a free-roaming dog (FRD). Mass vaccination campaigns against rabies and improving biosecurity measures to limit access of free-roaming dogs to farms can prevent the occurrence of RABV in dogs, livestock animals and personnel at risk in Nigeria.

Risk factors and molecular epidemiology of canine rabies in Beijing

Rabies is a serious public health problem in Asia and Africa. Approximately 99% of cases result from a bite by an infected dog. The high rabies risk countries are generally characterized by rapid urbanization; however, the prevalence and risk factors for rabies in urban dogs in developing countries remain unclear. Beijing, the capital of China, has been undergoing rapid urbanization, and implemented compulsory rabies immunization for dogs on October 1, 2014. We evaluated the effects of this immunization policy and investigated the dynamics and determinants of the spread of dog rabies virus (RABV) in Beijing. Between 2013 and 2018, the positive rate of rabies in biting dogs declined significantly; 79.6% of these were stray dogs. Reconstructed spatiotemporal diffusion for RABV presents a more central distribution within Beijing and a surrounding dissemination of virus lineage toward suburban areas, indicating that rabies is self-sustaining in Beijing. Continuous RABV phylogeographic reconstructions and logistic regression analysis of rabies cases revealed that rabies tended to remain in commercial areas, high-rise residences, demolition areas, and high-GDP, high-population and high-accessibility areas. Demolition areas, where many residents have abandoned their pets, are recognized as high-risk areas for rabies. Our findings reveal that the implementation of the immunization policy in Beijing has been effective in prevention and control of canine rabies, and because the prevalence of canine rabies results from multiple factors, emphasize the important role of multi-sectoral cooperation in rabies prevention and control.

Molecular characterization of rabies virus in trade dogs from Plateau state, Nigeria

Rabies is one of the most dreaded diseases known to mankind and it is endemic in Nigeria. This study was aimed at the detection and molecular characterisation of the rabies virus antigen in trade dogs in Plateau State. A cross-sectional study was performed to determine the prevalence of rabies antigen in the brain tissues of slaughtered dogs, from February to June 2018 using a direct fluorescent antibody test and mouse inoculation technique. Direct Fluorescent Antibody Test, Polymerase Chain Reaction (PCR), sequencing and phylogenetic analysis of the identified rabies virus were also undertaken. Phylogenetic analysis was done using molecular evolutionary genetic analysis (MEGA) 10 to determine their evolutionary relationship with other rabies virus sequences deposited in Genbank. Results indicated, out of 450 dog brain samples tested 14(3.1%) were positive for rabies virus antigen. Zoographic data obtained showed 66.3% of dogs were female, and the dogs were kept mainly for security (n=54) and breeding (n=28). Fourteen positive samples were used to inoculate 70 adult mice, only 8 samples were positive and 2 samples were also positive by RT PCR. Phylogenetic analysis of the nucleotide sequences showed that the two rabies virus sequences in this study, sequence MZ148311 had very close identity with EU038106 (99.8% homology) and sequence MZ148312 had perfect identity with EU038087 (100% homology) was most closely related to lyssavirus that has been reported to be circulating previously in Plateau and Nigeria, belonging to the Africa 2 lineage.

Impact and mitigation of sampling bias to determine viral spread: Evaluating discrete phylogeography through CTMC modeling and structured coalescent model approximations.

Bayesian phylogeographic inference is a powerful tool in molecular epidemiological studies, which enables reconstruction of the origin and subsequent geographic spread of pathogens. Such inference is, however, potentially affected by geographic sampling bias. Here, we investigated the impact of sampling bias on the spatiotemporal reconstruction of viral epidemics using Bayesian discrete phylogeographic models and explored different operational strategies to mitigate this impact. We considered the continuous-time Markov chain (CTMC) model and two structured coalescent approximations (Bayesian structured coalescent approximation [BASTA] and marginal approximation of the structured coalescent [MASCOT]). For each approach, we compared the estimated and simulated spatiotemporal histories in biased and unbiased conditions based on the simulated epidemics of rabies virus (RABV) in dogs in Morocco. While the reconstructed spatiotemporal histories were impacted by sampling bias for the three approaches, BASTA and MASCOT reconstructions were also biased when employing unbiased samples. Increasing the number of analyzed genomes led to more robust estimates at low sampling bias for the CTMC model. Alternative sampling strategies that maximize the spatiotemporal coverage greatly improved the inference at intermediate sampling bias for the CTMC model, and to a lesser extent, for BASTA and MASCOT. In contrast, allowing for time-varying population sizes in MASCOT resulted in robust inference. We further applied these approaches to two empirical datasets: a RABV dataset from the Philippines and a SARS-CoV-2 dataset describing its early spread across the world. In conclusion, sampling biases are ubiquitous in phylogeographic analyses but may be accommodated by increasing the sample size, balancing spatial and temporal composition in the samples, and informing structured coalescent models with reliable case count data.

Efficacy of Oral Rabies Vaccine Baits Containing SPBN GASGAS in Domestic Dogs According to International Standards.

(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.

Mathematical Analysis of Rabies Transmission Dynamics and Control

Rabies is a dangerous disease that kills many people than any other communicable disease and yet it is underrated. This results from the little knowledge on the myriad ways of transmission of the virus. A deterministic model is proposed to study the spread of the rabies virus in both domestic dogs (Canis familiaries) and humans (Homo sapiens). We elaborately studied the spread of the rabies virus from dogs to-dogs, dogs-to-humans and for the first time, humans-to-humans. Sensitivity analysis is performed to determine the influence of various parameters on the transmission of rabies the most. The rabies-free equilibrium and the endemic equilibrium points were determined and the conditions under which the equilibria are stable were also obtained. The stability conditions provide the conditions under which the disease will persist or get to be eradicated. Numerical solutions of the model were obtained using the ode45 routine in MATLAB. The study demonstrated that for rabies to be eradicated, the rate at which dogs are recruited must be decreased, culling of exposed and infected dogs should be increased and mass vaccination of the dog population should be targeted.

Rabies Virus Seroprevalence among Dogs in Limpopo National Park and the Phylogenetic Analyses of Rabies Viruses in Mozambique.

Rabies is considered a neglected disease among many developing Asian and African countries, including Mozambique, where its re-emergence is often attributed to low dog parenteral vaccination coverage. The objectives of this study were two-fold: (1) to assess the level of antibodies against rabies virus in dogs (n = 418) in Limpopo National Park (LNP), and (2) to genetically characterise selected rabies viruses from brain tissue samples collected in 2017 and 2018. To meet the first objective, we used the BioProTM Rabies blocking ELISA antibody kit, and the results were expressed as the percentage of blocking (%PB). Dog sera with PB ≥ 40% were considered positive for antibodies to rabies virus, whereas sera with PB < 40% were negative. Just under ninety percent (89.2%; n = 373) of dogs were seronegative, and the rest (10.8%; n = 45) had detectable levels of rabies virus-specific antibodies. All eight brain tissue samples were positive for rabies virus antigen using a direct fluorescent antibody test and amplified in a quantitative real-time PCR, but only five (n = 4 from dogs and n = 1 from a cat) were amplified in a conventional reverse-transcription PCR targeting partial regions of the nucleoprotein (N) and the glycoprotein (G) genes. All samples were successfully sequenced. Phylogenetically, the rabies viruses were all of dog origin and were very closely related to each other (Africa 1b rabies virus lineage). Furthermore, the sequences had a common progenitor with other rabies viruses from southern Africa, confirming the transboundary nature of rabies and the pivotal role of dogs in maintaining rabies cycles. The study demonstrates the principal application of the BioProTM rabies ELISA antibody for the detection of anti-lyssavirus-specific antibodies in the serum samples of dogs, and most importantly, it highlights the low levels of antibodies against rabies virus in this dog population.

A One Medicine Mission for an Effective Rabies Therapy.

Despite the disease's long history, little progress has been made toward a treatment for rabies. The prognosis for patient recovery remains dire. For any prospect of survival, patients require aggressive critical care, which physicians in rabies endemic areas may be reluctant or unable to provide given the cost, clinical expertise required, and uncertain outcome. Systematic clinical research into combination therapies is further hampered by sporadic occurrence of cases. In this Perspective, we examine the case for a One Medicine approach to accelerate development of an effective therapy for rabies through the veterinary care and investigational treatment of naturally infected dogs in appropriate circumstances. We review the pathogenesis of rabies virus in humans and dogs, including recent advances in our understanding of the molecular basis for the severe neurological dysfunction. We propose that four categories of disease process need to be managed in patients: viral propagation, neuronal degeneration, inflammation and systemic compromise. Compassionate critical care and investigational treatment of naturally infected dogs receiving supportive therapy that mimics the human clinical scenario could increase opportunities to study combination therapies that address these processes, and to identify biomarkers for prognosis and therapeutic response. We discuss the safety and ethics of this approach, and introduce the Canine Rabies Treatment Initiative, a non-profit organization with the mission to apply a One Medicine approach to the investigation of diagnostic, prognostic, and therapeutic options for rabies in naturally infected dogs, to accelerate transformation of rabies into a treatable disease for all patients.

Evolutionary analysis of rabies virus using the partial Nucleoprotein and Glycoprotein gene in Mumbai region of India.

Nearly 1.7 million cases of dog bites are reported every year in India and many cases of animal rabies are left unattended and undiagnosed. Therefore, a mere diagnosis of rabies is not sufficient to understand the epidemiology and the spread of the rabies virus (RV) in animals. There is a paucity of information about the evolutionary dynamics of RV in dogs and its biodiversity patterns in India. In total, 50 dog-brain samples suspected of rabies were screened by the nucleoprotein- (N) and glycoprotein- (G) gene PCR. The N and G genes were subsequently sequenced to understand the molecular evolution in these genes. The phylogenetic analysis of the N gene revealed that six isolates in the Mumbai region belonged to a single Arctic lineage. Time-scaled phylogeny by Bayesian coalescent analysis of the partial N gene revealed that the time to the most recent common ancestor (TMRCA) for the sequences belonged to the cluster from 2006.68 with a highest posterior density of 95 % betweeen 2005-2008, which is assigned to Indian lineage I. Migration pattern revealed a strong Bayes factor between Mumbai to Delhi, Panji to Hyderabad, Delhi to Chennai, and Chennai to Chandigarh. Phylogenetic analysis of the G gene revealed that the RVs circulating in the Mumbai region are divided into three lineages. Time-scaled phylogeny by the Bayesian coalescent analysis method estimated that the TMRCA for sequences under study was from 1993 and Indian clusters was from 1962. In conclusion, the phylogenetic analysis of the N gene revealed that six isolates belonged to single Arctic lineages along with other Indian isolates and they were clustered into a single lineage but divided into three clades based on the G-gene sequences. The present study highlights and enhances the current molecular epidemiology and evolution of RV and revealed strong location bias and geographical clustering within Indian isolates on the basis of N and G genes.

Detection of rabies virus antigen in brain tissue of dogs slaughtered for human consumption in Taraba State, Nigeria

Rabies as an ancient zoonosis constitutes a threat to public health by causing over 59,000 annual human mortalities worldwide. The aim of this study was to detect rabies virus in brain tissue of dogs slaughtered for human consumption in Taraba State, Nigeria. A total of 150 dogs comprising 136 adults and 14 puppies consisting of 82 males and 68 females was sampled from slaughter points in five Local Government Areas. Brain samples were collected from each dog in labeled sterile sample bottles and screened for rabies virus antigen using direct fluorescent antibody test (DFAT). Results showed that 3 out of the 150 (2%) brain samples screened were positive for rabies virus; out of which 2 were from Unguwan Kasa (14.3%) and 1 was from Quarter Five (7.1%). This therefore suggests the presence of rabies virus in dogs slaughtered for human consumption in Taraba State, Nigeria and their role as reservoirs of the virus. Therefore, there is need for awareness education on safe handling of dog meat to minimize the risk for butchers/meat handlers.&#x0D; Keywords: Brain samples, dogs, Prevalence, Rabies, Taraba State

Molecular study of rabies virus in slaughtered dogs in Billiri and Kaltungo local government areas of Gombe state, Nigeria

Rabies is one of the most dreadful diseases and a major viral zoonosis which has been shown to cause an almost 100% fatality rate in infected victims. It is characterized by acute progressive encephalitis in mammals. This study determined the genotypic characteristics of rabies virus in dogs slaughtered for human consumption based on sequence of a fragment of nucleoprotein gene. Brain tissues were collected from 50 dogs slaughtered in Billiri and Kaltungo Local Government Areas of Gombe State, Nigeria. Direct fluorescent antibody test (DFAT) was used to screen for the presence of rabies virus antigen. Viral RNA isolated from DFAT positive brain tissues were subjected to the reverse transcription polymerase chain reaction (RT-PCR) followed by sequencing of the amplicons. Maximum Likelihood (ML) was used to construct a phylogenetic tree for sequences obtained with 1000 bootstrap replicates. The DFAT detected rabies antigen in 3 (6%) of the 50 dog brain tissues, from which 1 (2%) was positive by RT-PCR. ML phylogeny approach of the nucleotide sequences inferred members as originating lyssavirus genus and dog species. Essentially, MK234794 in this study displayed 99.3% sequence similarity with other related rabies viruses in the Africa 2 cluster (Nigeria, Cameroon, Chad and Niger). Interestingly, MK234794 showed no cluster relation with the Africa 1a, 1b, 3 and Africa 4 clades, respectively. This indicates there is in-country and trans-boundary circulation of the rabies viruses with no co-circulation between the Africa lineages, especially as dogs are continuously being traded due to consumption of dog meat in West Africa. This finding has given additional insight into the molecular epidemiology of rabies virus in Nigeria, therefore providing more baseline information for future design of rabies control programs in the country.

Using phylogeographic approaches to analyse the dispersal history, velocity and direction of viral lineages-Application to rabies virus spread in Iran.

Recent years have seen the extensive use of phylogeographic approaches to unveil the dispersal history of virus epidemics. Spatially explicit reconstructions of viral spread represent valuable sources of lineage movement data that can be exploited to investigate the impact of underlying environmental layers on the dispersal of pathogens. Here, we performed phylogeographic inference and applied different post hoc approaches to analyse a new and comprehensive data set of viral genomes to elucidate the dispersal history and dynamics of rabies virus (RABV) in Iran, which have remained largely unknown. We first analysed the association between environmental factors and variations in dispersal velocity among lineages. Second, we present, test and apply a new approach to study the link between environmental conditions and the dispersal direction of lineages. The statistical performance (power of detection, false-positive rate) of this new method was assessed using simulations. We performed phylogeographic analyses of RABV genomes, allowing us to describe the large diversity of RABV in Iran and to confirm the cocirculation of several clades in the country. Overall, we estimate a relatively high lineage dispersal velocity, similar to previous estimates for dog rabies virus spread in northern Africa. Finally, we highlight a tendency for RABV lineages to spread in accessible areas associated with high human population density. Our analytical workflow illustrates how phylogeographic approaches can be used to investigate the impact of environmental factors on several aspects of viral dispersal dynamics.

Comparison of intra- and inter-host genetic diversity in rabies virus during experimental cross-species transmission.

The development of high-throughput genome sequencing enables accurate measurements of levels of sub-consensus intra-host virus genetic diversity and analysis of the role played by natural selection during cross-species transmission. We analysed the natural and experimental evolution of rabies virus (RABV), an important example of a virus that is able to make multiple host jumps. In particular, we (i) analyzed RABV evolution during experimental host switching with the goal of identifying possible genetic markers of host adaptation, (ii) compared the mutational changes observed during passage with those observed in natura, and (iii) determined whether the colonization of new hosts or tissues requires adaptive evolution in the virus. To address these aims, animal infection models (dog and fox) and primary cell culture models (embryo brain cells of dog and fox) were developed and viral variation was studied in detail through deep genome sequencing. Our analysis revealed a strong unidirectional host evolutionary effect, as dog-adapted rabies virus was able to replicate in fox and fox cells relatively easily, while dogs or neuronal dog cells were not easily susceptible to fox adapted-RABV. This suggests that dog RABV may be able to adapt to some hosts more easily than other host variants, or that when RABV switched from dogs to red foxes it lost its ability to adapt easily to other species. Although no difference in patterns of mutation variation between different host organs was observed, mutations were common following both in vitro and in vivo passage. However, only a small number of these mutations also appeared in natura, suggesting that adaptation during successful cross-species virus transmission is a complex, multifactorial evolutionary process.

Molecular characterization of the rabies virus' nucleoprotein in dogs from Arequipa, Peru

. To characterize the nucleoprotein (N) and establish the origin of the rabies virus in dogs coming from Arequipa. Thirty samples of nervous tissue from the departments of Arequipa and Puno were analyzed. Total RNA was extracted from the samples and cDNA was synthesized to amplify the nucleoprotein gene, sequence it, and perform bioinformatics analysis. . A defined group was formed with respect to the external group (European bat lyssavirus). This group was classified into two subgroups, one constituted by samples coming from Puno and Arequipa (subgroup A), and another one by samples from Puno (subgroup B), exhibiting a nucleotide identity of 99.9% in subgroup A. This group was classified in two subgroups, one constituted by samples coming from Puno and Arequipa (subgroup A), and another one by samples from Puno (subgroup B), observing a nucleotide identity of 99.9% in subgroup A. The groupings of viral sequences show that the cases of canine rabies reported in Arequipa are the result of the expansion of canine rabies from the endemic region of Puno.

Transmission dynamics of re-emerging rabies in domestic dogs of rural China.

Despite ongoing efforts to control transmission, rabies prevention remains a challenge in many developing countries, especially in rural areas of China where re-emerging rabies is under-reported due to a lack of sustained animal surveillance. By taking advantage of detailed genomic and epidemiological data for the re-emerging rabies outbreak in Yunnan Province, China, collected between 1999 and 2015, we reconstruct the demographic and dispersal history of domestic dog rabies virus (RABV) as well as the dynamics of dog-to-dog and dog-to-human transmission. Phylogeographic analyses reveal a lower diffusion coefficient than previously estimated for dog RABV dissemination in northern Africa. Furthermore, epidemiological analyses reveal transmission rates between dogs, as well as between dogs and humans, lower than estimates for Africa. Finally, we show that reconstructed epidemic history of RABV among dogs and the dynamics of rabid dogs are consistent with the recorded human rabies cases. This work illustrates the benefits of combining phylogeographic and epidemic modelling approaches for uncovering the spatiotemporal dynamics of zoonotic diseases, with both approaches providing estimates of key epidemiological parameters.

Low level of the immune response against rabies virus in dogs and cats, a cross-sectional study in sheltered animals, Santander, Colombia

ABSTRACT: Rabies is an incurable and fatal progressive viral encephalomyelitis that causes approximately 61,000 deaths a year globally mainly by dog bites, which show the importance of anti-rabies vaccination in dogs to achieve elimination of the disease. In Colombia, multiple cases of human rabies transmitted by dogs and cats have been reported since 1999, showing an increased significance of cats in the transmission of rabies, mainly in rabies of wild origin. Therefore, the aim of the present study was to evaluate the development of neutralizing antibodies in dogs and cats during the mass vaccination campaign of the second half of 2015 in the city of Bucaramanga. For this purpose, a descriptive cross-sectional study with convenience sampling was conducted in 382 dogs and cats (295 dogs, 87 cats), and an evaluation of the humoral immune response of the animals was performed by quantitative ELISA. The prevalence of optimal neutralizing antibodies (&gt;0.5 IU/ml) was only 32.76% (95% CI=28.05-37.46%) in the entire population studied and most of the animals did not have an adequate response to the vaccination, or seroconversion was not detected on them (65.45-95% confidence interval, CI=60.68-70.21%). Significant difference was found between the neutralizing antibody titers in cats and dogs, with a higher neutralizing response in cats. In conclusion, although mass vaccination campaigns for dogs and cats are the most important measure to interrupt virus circulation among the animals, achievement of a good neutralizing immune response in the animals is useful to demonstrate that vaccination has been successful, allowing the maintenance of the required minimum levels of population immunity. These results will allow the implementation of corrective measures in Bucaramanga to achieve better seroconversion rates. Other cities are expected to implement similar seroconversion assessments to verify the quality of effective anti-rabies vaccination in animal populations.

Rabies Virus.

This infographic describes the transmission cycle of rabies virus in domestic dogs and the necessity of a One Health approach, integrating medical and veterinary interventions, to control and eliminate human rabies deaths. Rabies virus (RABV) causes an acute, fatal neurological infection in humans and other mammals, transmitted through the saliva of rabid animals via a bite or scratch. From the site of infection the virus travels along neurons to the central nervous system (CNS), where viral replication leads to symptoms and systemic spread. Once symptomatic, the disease is nearly 100% fatal. However, the disease is 100% vaccine-preventable through the prompt administration of human postexposure prophylaxis (PEP) and vaccination of animal reservoirs. While RABV has a broad host range, domestic dogs cause over 99% of all human cases, killing 59000 people every year. Human PEP is costly (US$11-150 per dose) and often difficult to obtain. Dog vaccination is a considerably more cost-effective and feasible method to reduce the incidence of human rabies. With this in mind, the World Health Organisation (WHO) and partners have set a target for the global elimination of dog-mediated human rabies, through control of the disease in dogs, by 2030.

Development of a Blocking ELISA for Measuring Rabies Virus-specific Antibodies in Animals

Rabies virus (RABV)-specific antibodies in animals and humans are measured using standard methods such as fluorescent antibody virus neutralization (FAVN) tests and rapid fluorescent focus inhibition tests, which are based on cell culture systems. An alternative assay that is safe and easy to perform is required for rapid sero-surveillance following mass vaccination of animals. Two purified monoclonal antibodies (4G36 and B2H17) against RABV were selected as capture and detection antibodies, respectively. A genetically modified RABV, the ERAGS strain, was propagated and concentrated by polyethylene glycol precipitation. Optimal conditions for the RABV antigen, antibodies, and serum dilution for a blocking enzymelinked immune sorbent assay (B-ELISA) were established. We evaluated the sensitivity, specificity, and accuracy of the B-ELISA using serum samples from 138 dogs, 71 raccoon dogs, and 25 cats. The B-ELISA showed a diagnostic sensitivity of 95.8-96.3%, specificity of 91.3-100%, and accuracy of 96.0-97.2% compared to the FAVN test. These results suggest that the B-ELISA is useful for sero-surveillance of RABV in dogs, raccoon dogs, and cats.