What would be the impact on the rabies risk of reducing the waiting period before dogs are imported? A modelling study based on the European Union legislation.

AIMS: Lyssavirus rabies (RABV) is responsible for a major zoonotic infection that is almost always lethal once clinical signs appear. Rabies can be (re)introduced into rabies-free areas through transboundary dog movements, thus compromising animal and human health. A number of measures have been implemented to prevent this happening, one of which is the waiting period (WP) after anti-rabies vaccination and serological testing. This WP ensures that antibodies assessed through the serological test are due to the vaccine, not to infection. Indeed, if antibodies are due to RABV infection, the dog should display clinical signs within this WP and would not therefore be imported. METHODS AND RESULTS: Within a framework of quantitative risk assessment, we used modelling approaches to evaluate the impact of this WP and its duration on the risk of introducing rabies via the importation of dogs into the European Union. Two types of models were used, a classical stochastic scenario tree model and an individual-based model, both parameterised using scientific literature or data specifically applicable to the EU. Results showed that, assuming perfect compliance, the current 3-month waiting period was associated with a median annual number of 0.04 infected dogs imported into the EU. When the WP was reduced, the risk increased. For example, for a 1-month WP, the median annual number of infected dogs imported was 0.17 or 0.15 depending on the model, which corresponds to a four-fold increase. CONCLUSION: This in silico study, particularly suitable for evaluating rare events such as rabies infections in rabies-free areas, provided results that can directly inform policymakers in order to adapt regulations linked to rabies and animal movements.

Retrospective analysis of sero-prevalence and bait uptake estimations in foxes after oral rabies vaccination programmes at European level: Lessons learned and paths forward.

Rabies caused by the Classical Rabies Virus (Lyssavirus rabies abbreviated RABV) in the European Union has been close to elimination mainly thanks to Oral Rabies Vaccination (ORV) campaigns targeting wildlife (primarily red foxes). ORV programmes co-financed by the European Commission include a monitoring-component to assess the effectiveness of the ORV campaigns at national level. This assessment is performed by a random collection of red foxes in the vaccinated areas with control of antibodies presence by serological analysis and control of bait uptake by detection of biomarkers (tetracycline incorporated into the baits) in the bones and teeth. ORV programmes aim to a vaccine coverage high enough to immunize (ideally) 70 % of the reservoir population to control the spread of the disease. European Union (EU) programmes that led to almost elimination of rabies on the territory have been traditionally found to have a bait uptake average of 70 % (EU countries; 2010-2020 period) while the seroconversion data showed an average level of 40 % (EU countries; 2010-2020 period). To better understand variations of these indicators, a study was been set up to evaluate the impact of several variables (linked to the vaccination programme itself and linked to environmental conditions) on the bait uptake and the seroconversion rate. Thus, pooling data from several countries provides more powerful statistics and the highest probability of detecting trends. Results of this study advocate the use of a single serological test across the EU since data variation due to the type of test used was higher than variations due to field factors, making the interpretation of monitoring results at EU level challenging. In addition, the results indicates a negative correlation between bait uptake and maximum temperatures reached during ORV campaigns questioning the potential impact of climatic change and associated increase of temperatures on the ORV programmes efficiency. Several hypotheses requesting additional investigation are drawn and discussed in this paper.

Assessment of virus and Leptospira carriage in bats in France.

With over 1,400 species worldwide, bats represent the second largest order of mammals after rodents, and are known to host major zoonotic pathogens. Here, we estimate the presence of pathogens in autochthonous bat populations. First, we set out to check our samples for PCR amplification efficiency by assessing the occurrence of inhibited PCR reactions from different types of bat samples with amplifying the housekeeping gene ß-actin. Second, we investigated the presence of five targeted pathogens in a French bat population using PCR. We targeted viral RNA of Canine distemper virus, Alphacoronavirus, Lyssavirus, Rotavirus and bacterial Leptospira DNA. To do so, we screened for these viruses in bat faecal samples as well as in oropharyngeal swab samples. The presence of Leptospira was assessed in urine, kidney, lung and faecal samples. Results showed a frequency of inhibited reactions ranging from 5 to 60% of samples, varying according to the sample itself and also suspected to vary according to sampling method and the storage buffer solution used, demonstrating the importance of the sampling and storage on the probability of obtaining negative PCR results. For pathogen assessment, rotavirus and alphacoronavirus RNA were detected in Myotis myotis, Myotis daubentonii, Myotis emarginatus and Rhinolophus ferrumequinum bats. Rotaviruses were also detected in Barbastella barbastellus. The presence of alphacoronavirus also varied seasonally, with higher frequencies in late summer and October, suggesting that juveniles potentially play an important role in the dynamics of these viruses. Leptospira DNA was detected in M. myotis and M. daubentonii colonies. The 16S rRNA sequences obtained from Leptospira positive samples showed 100% genetic identity with L. borgpetersenii. Neither canine distemper virus nor lyssavirus RNA were detected in any of the tested samples. This study is the first to show the presence of Leptospira in autochthonous French bats in addition to coronavirus and rotavirus RNA previously reported in European autochthonous bats.

Whole genome sequencing and phylogenetic characterisation of rabies virus strains from Moldova and north-eastern Romania.

BACKGROUND: Rabies is the oldest fatal zoonotic disease recognised as a neglected tropical disease and is caused by an RNA virus belonging to the genus Lyssavirus, family Rhabdoviridae. METHODOLOGY/PRINCIPAL FINDINGS: A deep molecular analysis was conducted on full-length nucleoprotein (N) gene and whole genome sequences of rabies virus from 37 animal brain samples collected between 2012 and 2017 to study the circulation of rabies virus (RABV) variants. The overall aim was to better understand their distribution in Moldova and north-eastern Romania. Both Sanger and high throughput sequencing on Ion Torrent and Illumina platforms were performed. Phylogenetic analysis of the RABV sequences from both Moldova and Romania revealed that all the samples (irrespective of the year of isolation and the species) belonged to a single phylogenetic group: north-eastern Europe (NEE), clustering into three assigned lineages: RO#5, RO#6 and RO#7. CONCLUSIONS/SIGNIFICANCE: High throughput sequencing of RABV samples from domestic and wild animals was performed for the first time for both countries, providing new insights into virus evolution and epidemiology in this less studied region, expanding our understanding of the disease.

Quantitative risk assessment of rabies being introduced into mainland France through worldwide noncommercial dog and cat movements.

France has been rabies-free among nonflying mammals since 2001. Despite this status, the rabies virus has been introduced several times through noncommercial pet movements, posing a threat of infection by this 100%-lethal zoonosis among local animal and human populations. To quantify the risk of rabies being introduced through worldwide noncommercial dog and cat movements, we performed a quantitative risk assessment using stochastic scenario tree modeling. The mean annual probability of at least one rabies introduction incident was 0.35 (median: 0.24, 90% prediction interval (PI) [0.04; 0.98]) and the mean annual number of rabies-infected pets introduced through pet movements was 0.96 (median: 0.27, 90% PI [0.04; 3.88]). These results highlight a nonnegligible, even high risk due to the associated consequences of such events. In alternative scenario testing, preventive anti-rabies vaccination proved to be an effective measure since removing the vaccination requirement led to a > 15-fold increase in risk. The serological testing requirement had less of an effect (approximately two-fold increase when removed) and the posttest waiting period to ensure that antibodies were not linked to an infection had a negligible effect. Any change in pet owner compliance, especially regarding vaccination, could have a major impact on the risk. This study also shows that reinforced border control staff training could be more effective in reducing risk than more frequent checks. These results provide quantitative data for assessing the probability of the rabies virus entering France, and could help policymakers decrease this risk in rabies-free areas.

Risks related to a possible reduction of the waiting period for dogs after rabies antibody titration to 30 days compared with 90 days of the current EU legislative regime.

EFSA received a mandate from the European Commission to assess the risks related to a possible reduction of the waiting period after rabies antibody titration test to 30 days compared with 90 days of the current EU legislation, for dogs moving from certain non-EU countries to the EU. This Scientific Report assessed the probability of introduction of rabies into the EU through commercial and non-commercial movements of vaccinated dogs with a positive titration test (≥ 0.5 IU/mL) if the waiting period decreases from 90 to 30 days. Assuming that all the legal requirements are complied with, the risk of transmission of rabies through the movement of a vaccinated dog is related to the risk of introducing an animal incubating rabies that was infected before the day of vaccination or shortly after vaccination but before the development of immunity (21 days post-vaccination). Using published data on the incubation period for experimental and field cases in dogs and considering the rabies incidence data in certain countries, the aggregated probability for the annual introduction of rabies through dogs was assessed. Considering the uncertainty related to the duration of the incubation period, the number of imported dogs, and the disease incidence in some countries it was concluded with a 95% certainty that the maximum number of rabies-infected imported dogs complying with the regulations in a 20-year period could increase from 5 to 20 when decreasing the waiting period from 90 to 30 days. Nevertheless, the potential impact of even a small increase in probability means the risk is increased for a region like the EU where rabies has long been a focus for eradication, to protect human and animal health.

Genetic identification of bat species for pathogen surveillance across France.

With more than 1400 chiropteran species identified to date, bats comprise one-fifth of all mammalian species worldwide. Many studies have associated viral zoonoses with 45 different species of bats in the EU, which cluster within 5 families of bats. For example, the Serotine bats are infected by European Bat 1 Lyssavirus throughout Europe while Myotis bats are shown infected by coronavirus, herpesvirus and paramyxovirus. Correct host species identification is important to increase our knowledge of the ecology and evolutionary pattern of bat viruses in the EU. Bat species identification is commonly determined using morphological keys. Morphological determination of bat species from bat carcasses can be limited in some cases, due to the state of decomposition or nearly indistinguishable morphological features in juvenile bats and can lead to misidentifications. The overall objective of our study was to identify insectivorous bat species using molecular biology tools with the amplification of the partial cytochrome b gene of mitochondrial DNA. Two types of samples were tested in this study, bat wing punches and bat faeces. A total of 163 bat wing punches representing 22 species, and 31 faecal pellets representing 7 species were included in the study. From the 163 bat wing punches tested, a total of 159 were genetically identified from amplification of the partial cyt b gene. All 31 faecal pellets were genetically identified based on the cyt b gene. A comparison between morphological and genetic determination showed 21 misidentifications from the 163 wing punches, representing ~12.5% of misidentifications of morphological determination compared with the genetic method, across 11 species. In addition, genetic determination allowed the identification of 24 out of 25 morphologically non-determined bat samples. Our findings demonstrate the importance of a genetic approach as an efficient and reliable method to identify bat species precisely.

Management practices of dog and cat owners in France (pet traveling, animal contact rates and medical monitoring): Impacts on the introduction and the spread of directly transmitted infectious pet diseases.

A number of owner practices among the pet dog and cat population can influence the dynamics of directly transmitted infectious dog and cat diseases, including zoonotic ones. To better depict these management practices, which include pet traveling, contact rates with other companion animals and their medical monitoring (which herein includes prevention aspects), we surveyed 2,122 dog- and/or cat-owning French households through an anonymous online questionnaire. Trips with dogs within the European Union (EU) were frequent, while cats travelled less frequently within the EU and both cats and dogs travelled less frequently outside the EU. Recurrent illegal trips with dogs and cats (non-compliant with regulatory measures) were observed in a context of non-systematic pet border controls. We found that a large proportion of dogs are taken for walks in metropolitan France, with frequent intraspecific contacts (1.4 contacts/day on average), but only a minority (1.4%) of dogs were allowed to roam freely. On the other hand, 59.7% of cat owners allowed their cats to roam freely. We classified pet owners according to different profiles, some of which may be considered 'at risk' for directly transmitted infectious pet diseases. Indeed, one dog owner profile and one cat owner profile depict 'spreaders' of pet diseases (high connectivity with other individuals, little medical monitoring but no traveling) and another dog owner profile describes a potential 'introducer' and 'spreader' of pet diseases (foreign travel, high connectivity with other individuals, and intermediate medical monitoring). While these 'at risk' profiles represent only a minority of French pet owners, they should be better characterized to reinforce targeted prevention designed to minimize the risk of (re)introduction and (re)emergence of directly transmitted infectious dog and cat diseases in France, especially when considering zoonoses with a significant potential impact, such as rabies.

Benefit-Risk Assessment of the French Surveillance Protocol of Apparently Healthy Biting Dogs and Cats for Human Rabies Prevention.

In France, apparently healthy dogs and cats that bite humans must undergo an observation period of 15 days with three veterinary visits to ascertain that they remain healthy, indicating that no zoonotic transmission of rabies virus occurred via salivary presymptomatic excretion. This surveillance protocol is mandatory for all pets that have bitten humans, despite France's rabies-free status in non-flying mammals (i.e., a very low rabies risk). In this context, we aimed to perform a benefit-risk assessment of the existing regulatory surveillance protocol of apparently healthy biting animals, as well as alternative surveillance protocols. A scenario-tree modelling approach was used to consider the possible successions of events between a dog or cat bite and a human death attributed to either rabies or to lethal harm associated with the surveillance protocol (e.g., lethal traffic accidents when traveling to veterinary clinics or anti-rabies centers). The results demonstrated that the current French surveillance protocol was not beneficial, as more deaths were generated (traffic accidents) than avoided (by prompt post-exposure prophylaxis administration). We showed here that less stringent risk-based surveillance could prove more appropriate in a French context. The results in this study could allow policy-makers to update and optimize rabies management legislation.

Multi-annual performance evaluation of laboratories in post-mortem diagnosis of animal rabies: Which techniques lead to the most reliable results in practice?

Rabies diagnosis proficiency tests on animal specimens using four techniques (FAT, RTCIT, conventional RT-PCR and real-time RT-PCR) were organised over 10 years (2009-2019). Seventy-three laboratories, of which 59% were from Europe, took part. As the panels were prepared with experimentally-infected samples, the error rate of laboratories on positive and negative samples was accurately estimated. Based on fitted values produced by mixed modelling including the variable "laboratory" as a random variable to take into account the longitudinal design of our dataset, the technique that provided the most concordant results was conventional RT-PCR (99.3%; 95% CI 99.0-99.6), closely followed by FAT (99.1%; 95% CI 98.7-99.4), real-time RT-PCR (98.7%; 95% CI 98.1-99.3) and then RTCIT (96.8%; 95% CI 95.8-97.7). We also found that conventional RT-PCR provided a better diagnostic sensitivity level (99.3% ±4.4%) than FAT (98.7% ±1.6%), real-time RT-PCR (97.9% ±0.8%) and RTCIT (95.3% ±5.1%). Regarding diagnostic specificity, RTCIT was the most specific technique (96.4% ±3.9%) followed closely by FAT (95.6% ±3.8%), real-time RT-PCR (95.0% ±1.8%) and conventional RT-PCR (92.9% ±0.5%). Due to multiple testing of the samples with different techniques, the overall diagnostic conclusion was also evaluated, and found to reach an inter-laboratory concordance level of 99.3%. The concordance for diagnostic sensitivity was 99.6% ±2.0% and for diagnostic specificity, 98.0% ±8.5%. Molecular biology techniques were, however, found to be less specific than expected. The potential reasons for such findings are discussed herein. The regular organisation of performance tests has contributed to an increase in the performance of participating laboratories over time, demonstrating the benefits of such testing. Maintaining a high-quality rabies diagnosis capability on a global scale is key to achieving the goal of eliminating dog-mediated human rabies deaths. The regular organisation of exercises on each continent using selected local strains to be tested according to the local epidemiological situation is one factor that could help increase reliable diagnosis worldwide. Rabies diagnosis capabilities could indeed be enhanced by providing adequate and sustainable proficiency testing on a large scale and in the long term.

Evaluation of the Worldwide Occurrence of Rabies in Dogs and Cats Using a Simple and Homogenous Framework for Quantitative Risk Assessments of Rabies Reintroduction in Disease-Free Areas through Pet Movements.

Dog and cat rabies cases imported from rabies enzootic countries represent a major threat for areas that have acquired rabies-free status and quantitative risk analyses (QRAs) are developed in order to assess this risk of rabies reintroduction through dog and cat movements. Herein we describe a framework to evaluate dog and cat rabies incidence levels in exporting countries along with the associated uncertainty for such QRAs. For enzootic dog rabies areas (EDRAs), we extended and adapted a previously published method to specify the relationship between dog rabies vaccination coverage and canine rabies incidence; the relationship between dog and cat rabies incidences; and then to predict annual dog and cat rabies incidences. In non-enzootic dog rabies areas (nEDRAs), we provided annual incidence based on declared dog and cat rabies cases. For EDRAs, we predicted an annual incidence potentially greater than 1.5% in dogs and about ten times lower in cats with a high burden in Africa and Asia but much lower in Latin America. In nEDRAs, the occurrence of rabies was lower and of similar magnitude in dogs and cats. However, wildlife could still potentially infect dogs and cats through spillover events. This framework can directly be incorporated in QRAs of rabies reintroduction.

Zero Endemic Cases of Wildlife Rabies (Classical Rabies Virus, RABV) in the European Union by 2020: An Achievable Goal.

The elimination of rabies transmitted by Classical Rabies Virus (RABV) in the European Union (EU) is now in sight. Scientific advances have made it possible to develop oral vaccination for wildlife by incorporating rabies vaccines in baits for foxes. At the start of the 1980s, aerial distribution of vaccine baits was tested and found to be a promising tool. The EU identified rabies elimination as a priority, and provided considerable financial and technical resources to the infected EU Member States, allowing regular and large-scale rabies eradication programs based on aerial vaccination. The EU also provides support to non-EU countries in its eastern and south eastern borders. The key elements of the rabies eradication programs are oral rabies vaccination (ORV), quality control of vaccines and control of their distribution, rabies surveillance and monitoring of the vaccination effectiveness. EU Member States and non-EU countries with EU funded eradication programs counted on the technical support of the rabies subgroup of the Task Force for monitoring disease eradication and of the EU Reference Laboratory (EURL) for rabies. In 2018, eight rabies cases induced by classical rabies virus RABV (six in wild animals and two in domestic animals) were detected in three EU Member States, representing a sharp decrease compared to the situation in 2010, where there were more than 1500 cases in nine EU Member States. The goal is to reach zero cases in wildlife and domestic animals in the EU by 2020, a target that now seems achievable.

An inter-laboratory comparison to evaluate the technical performance of rabies diagnosis lateral flow assays.

As in previous years, the European Union Reference Laboratory (EURL) for rabies organised in 2018 an Inter-laboratory trial (ILT) on rabies diagnosis. Contrarily to past years, the 2018 ILT did not aim to evaluate the performance of participating laboratories, but the technical performance of new rapid tests. Two lateral Flow Assays (LFA), namely the Anigen® and the CDIA™ Rabies Virus Antigen Rapid Test" (commercialized by Bionote and Creative Diagnostics Cie respectively), were evaluated together with the Fluorescent Antibody Test (FAT). One panel of virus samples (including RABV as well as EBLV1a, EBLV-1b, and EBLV2 strains) was sent to participating laboratories to compare results obtained with these different techniques. The study revealed that the FAT provided a good agreement toward expected results for both negative/positive samples (99.1%). The Anigen® test produced similar results to the FAT, with only one false negative result (0.5%) reported by all participants and a concordance of 100% for all but one sample demonstrating a good inter-laboratory reproducibility of the Anigen® batch. The CDIA™ test produced reproducible results for Rabies Virus (RABV) samples only. However, it hardly detected the Bokeloh Bat Lyssavirus (BBLV) and the European Bat Lyssaviruses types 1b and 2 (EBLV-1b and EBLV-2) in most laboratories resulting in a moderate inter-laboratory concordance (58.4%-82.7%) for these lyssaviruses. The two LFAs provided reliable and reproducible results on all RABV samples (100%) but lead to heterogeneous performances with other lyssaviruses leading to different levels of diagnostic/analytical sensitivity, specificity. The study confirmed that LFAs should be used with caution and that their validation are of upmost importance before any use in laboratories.

In-depth genome analyses of viruses from vaccine-derived rabies cases and corresponding live-attenuated oral rabies vaccines.

Live-attenuated rabies virus strains such as those derived from the field isolate Street Alabama Dufferin (SAD) have been used extensively and very effectively as oral rabies vaccines for the control of fox rabies in both Europe and Canada. Although these vaccines are safe, some cases of vaccine-derived rabies have been detected during rabies surveillance accompanying these campaigns. In recent analysis it was shown that some commercial SAD vaccines consist of diverse viral populations, rather than clonal genotypes. For cases of vaccine-derived rabies, only consensus sequence data have been available to date and information concerning their population diversity was thus lacking. In our study, we used high-throughput sequencing to analyze 11 cases of vaccine-derived rabies, and compared their viral population diversity to the related oral rabies vaccines using pairwise Manhattan distances. This extensive deep sequencing analysis of vaccine-derived rabies cases observed during oral vaccination programs provided deeper insights into the effect of accidental in vivo replication of genetically diverse vaccine strains in the central nervous system of target and non-target species under field conditions. The viral population in vaccine-derived cases appeared to be clonal in contrast to their parental vaccines. The change from a state of high population diversity present in the vaccine batches to a clonal genotype in the affected animal may indicate the presence of a strong bottleneck during infection. In conclusion, it is very likely that these few cases are the consequence of host factors and not the result of the selection of a more virulent genotype. Furthermore, this type of vaccine-derived rabies leads to the selection of clonal genotypes and the selected variants were genetically very similar to potent SAD vaccines that have undergone a history of in vitro selection.

Reconsidering Oral Rabies Vaccine Bait Uptake Evaluation at Population Level: A Simple, Noninvasive, and Ethical Method by Fecal Survey Using a Physical Biomarker.

Tetracycline and rhodamine are bait uptake biomarkers commonly used for decades in oral rabies vaccination campaigns. They require tooth collection and the capture or death of animals. Here, we considered the use of marked baits with plastic beads, a physical biomarker allowing noninvasive scat survey by direct observation in the field. Such methodology would be in compliance with animal welfare concerns. The development of a relative bait uptake estimation on the basis of observation marked scat could indeed be beneficial for the monitoring of oral vaccination programs, especially in programs dedicated to dogs ( Canis familiaris) or protected species.

Lleida Bat Lyssavirus isolation in Miniopterus schreibersii in France.

Bat rabies cases are attributed in Europe to five different Lyssavirus species of 16 recognized Lyssavirus species causing rabies. One of the most genetically divergent Lyssavirus spp. has been detected in a dead Miniopterus schreibersii bat in France. Brain samples were found positive for the presence of antigen, infectious virus and viral RNA by classical virological methods and molecular methods respectively. The complete genome sequence was determined by next-generation sequencing. The analysis of the complete genome sequence confirmed the presence of Lleida bat lyssavirus (LLEBV) in bats in France with 99.7% of nucleotide identity with the Spanish LLEBV strain (KY006983).

Evaluation of six TaqMan RT-rtPCR kits on two thermocyclers for the reliable detection of rabies virus RNA.

Rabies is diagnosed postmortem in animals, based on tests prescribed by the World Organization for Animal Health (OIE), such as the fluorescent antibody test, the direct rapid immunohistochemistry test, or pan-lyssavirus PCR assays. Several reverse-transcription real-time PCR (RT-rtPCR) methods have been developed and validated for rapid and accurate detection of lyssaviruses. We evaluated the performance of 6 TaqMan RT-rtPCR kits using different commercial master mixes and 2 real-time thermocyclers. Changing the master mix overall did not influence the TaqMan RT-rtPCR performance, regardless of the thermocycler used. The limits of detection at the 95% confidence level were 18.1-25.8 copies/µL for the Rotor-Gene Q MDx thermocycler and 16.7-21.5 for the Mx3005P thermocycler. Excellent repeatability was demonstrated for rabies virus (RABV) RNA samples of 100, 50, and 25 copies/µL regardless of the thermocycler used. RABV field samples ( n = 35) isolated worldwide gave positive results using the most efficient of the 6 kits tested, with a copy number of 6.03 × 102 to 6.78 × 107 RNA copies per reaction. The TaqMan RT-rtPCR assay provides sensitive and rapid amplification of RABV RNA.

Control and elimination of rabies in Croatia.

Despite the implementation of control measures (preventive dog vaccination), rabies has become endemic in Croatia, with red foxes being the main reservoir species. Oral rabies vaccination (ORV) campaigns supported by the European Commission have been conducted twice a year since the spring of 2011. The first campaigns were limited to the northern and eastern parts of the country, and from the autumn of 2012, the program was extended to the entire country. The Lysvulpen vaccine containing the SAD Bern strain was used for ORV. Following the vaccination campaigns, the number of rabies cases decreased, and the last positive case was recorded in February 2014. The bait uptake ranged from 24.86% to 84.62% and the immunisation rate from 11.24% to 35.64%.

Correction to: Oral vaccination of dogs: a well-studied and undervalued tool for achieving human and dog rabies elimination.

The original article [1] contained an error in the Author details paragraph. "5Neglected Zoonotic Diseases, World Health Organization, Geneva, Switzerland" should be replaced by "5Le Grand-Saconnex, Switzerland".

Oral vaccination of dogs: a well-studied and undervalued tool for achieving human and dog rabies elimination.

The mass vaccination of dogs is a proven tool for rabies prevention. Besides parenteral delivery of inactivated vaccines, over the past several decades, several self-replicating biologics, including modified-live, attenuated and recombinant viruses, have been evaluated for the oral vaccination of dogs against rabies. Vaccines are included within an attractive bait for oral consumption by free-ranging dogs. Due to the high affinity between dogs and humans, such biologics intended for oral vaccination of dogs (OVD) need to be efficacious as well as safe. Baits should be preferentially attractive to dogs and not to non-target species. Although many different types have been evaluated successfully, no universal bait has been identified to date. Moreover, high bait acceptance does not necessarily mean that vaccine efficacy and programmatic success is predictable. The use of OVD in the laboratory and field has demonstrated the safety and utility of this technology. Within a One Health context, OVD should be considered as part of a holistic plan for the global elimination of canine rabies.