Although conventional immunohistochemistry for neurotropic Rabies virus (RABV) usually shows a high preference for neurons, non-neuronal cells are also potential target cells and abortive infection of astrocytes is considered a main trigger of innate immunity in the CNS. While in vitro studies indicated differences between field and less virulent lab-adapted RABVs, a systematic and quantitative comparison of astrocyte tropism in vivo is lacking. Here, a recently developed solvent-based tissue clearing technique was used to measure the RABV cell tropism in infected brains. Immunofluorescence analysis of 1 mm-thick tissue slices enabled 3D segmentation and quantification of infection frequencies of astrocytes and neurons. Comparison of highly virulent street virus clones from fox, dog, and raccoon with three lab strains of intermediate and low virulence revealed remarkable differences in the ability to infect astrocytes in vivo. While all viruses and infection routes led to comparable neuron infection frequencies, striking differences were detected for the infection of astrocytes. Consistent and inoculation route-independent astrocyte infection by field viruses, together with route-dependent or undetectable astrocyte infection by lab-adapted or vaccine viruses strongly suggests a model in which the ability to establish productive astrocyte infection in vivo functionally distinguishes field and attenuated lab RABV strains.

Aims: To review canine rabies, mass parental vaccination, human post-exposure prophylaxis, gene therapy and costs for fighting rabies. Place and Duration of Study: Department of Animal Science – Other, Nelwan Institution for Human Resource Development, Indonesia, between December 2017 and March 2018. Methodology: The author searched the Pubmed Database at NCBI for articles on rabies disease published between 2007 and 2018. All articles were open access and in English. For rabies virus examination, Seller’s test was used. In this article, references written by the author and other relevant publications were included. The author reviewed a rabies dog case kept at Nelwan Institution for Human Resource Development. Results: The dog showed clinical signs such as inappetance, urinary frequency and soaking in a small, juicy drain. Currently, to treat rabies, no drugs are available. For rabies prevention, vaccination is the best way. To eradicate rabies, mass vaccination in dogs, post-exposure prophylaxis, and gene therapy should be used. Fort rabies disease eradication, minimum of 70% of the dog population should receive the vaccination. In addition, humans with category II exposure should receive a rabies vaccine and rabies immunoglobulin. Conclusion: To eradicate rabies, vaccinations are required. In addition, gene therapy can eliminate rabies from the infected neurons by using rAAV-N796. CRISPR/Cas9 system in combination with the MMEJ-based method. Furthermore, mass parental vaccination, post-exposure prophylaxis, and gene therapy can reduce costs in controlling rabies disease.

The growing population in Yucatan has led to the expansion of construction in the Mayan jungle for tourist spaces, residential areas, and agriculture. Recently, rabies cases in cats (Felis catus) have increased. This study aimed to perform the antigenic and genetic characterization of the RABV and to try to identify factors contributing to the increased transmission of rabies virus from wildlife to cats in the State of Yucatan. The ArcGIS software was employed to generate maps depicting the geographic locations of rabies outbreaks in cats. A mixed-effects Poisson regression model was applied to investigate the association between cat rabies cases and sociodemographic and ecological variables. A total of nine feline rabies cases occurred, three antigenic variants were detected: RVV1 (n=1); (RVV3) (n=1), and the canine-originated atypical variant (n=7). Rabies cases were spatially distributed at the urban-jungle interface, which may favor contact between cats and wildlife. No statistically significant associations with sociodemographic/ecological variables were observed. Urban areas with low (0-2%) to moderate (2-4%) changes in land cover showed a substantial increase in the probability of feline rabies cases. This study highlights the concerning resurgence of rabies infections in cats, emphasizing the looming threat of its reintroduction in dogs should vaccination rates diminish. The genetic affinity between the atypical variant and the canine virus underscores the urgent need for vigilance in maintaining high vaccination coverage across all susceptible species.

Rabies is a viral zoonotic disease caused by the lyssavirus and endemic to most tropical and sub-tropical areas of the world. This study reports on the occurrence of rabies in dogs and relates the results to the vaccination status of the dogs. A total of each of the 50 brain and 50 saliva samples from 100 dogs were tested for Lyssavirus using RT-PCR and visualized using gel electrophoresis. Additionally, a histopathological diagnosis of rabies was made on the brain samples. A lyssavirus positivity of 34 % was realized with 29/50 from brain and 5/50 from saliva being positive for rabies. Statistical significance was associated with vaccination (p=0.011) and health (p=0.0) statuses and attack by a stray dog prior to exhibition of signs of rabies (p=0.002). The rabid positive dogs had bitten at least 15 humans and a dog before their demise. This study confirms vaccination as an effective tool for the prevention of rabies. However, elimination of the disease is possible if there is an established and effective collaboration between veterinary and human medicine through one health.

Rabies remains a significant global threat, yet accurate estimations of its impact are hindered by the lack of confirmatory diagnoses. Postmortem confirmation of rabies traditionally involves invasive brain tissue testing, a process met with resistance from deceased patients' families, impeding consent. This paper presents and evaluates an innovative yet unpublished transnasal approach for postmortem brain tissue collection, offering a minimally invasive, easier, faster, and safer method. This method preserves the cadaver's integrity, potentially easing family reluctance towards autopsies. The limited testing of both human and animal rabies in Ghana highlights the challenges in diagnosing this fatal disease. Scarce diagnostic resources and the complexity of obtaining brain tissue samples exacerbate the issue. Cultural and religious beliefs surrounding autopsies contribute to familial hesitation, viewing these procedures as disruptive and disfiguring, further complicating consent. The transnasal technique involves approaching the brain tissue through the nostrils and cribriform plate without any superficial manipulation of the patient's head and face, thereby preserving aesthetics and natural features of the person. Technological advancements and seamless One Health collaboration among governmental, non-governmental, and research entities locally and globally culminated in Ghana's first confirmed rabies diagnosis using this method of brain tissue collection. This success emphasizes the efficiency and feasibility of transnasal brain collection approach and the invaluable role of the One Health approach and collaborative efforts in overcoming diagnostic challenges in rabies control.

Rabies persists as a longstanding issue in Zambia, despite being preventable. The current control measures, including dog vaccination, population control, and movement restriction, guided by ‘The Control of Dogs Act Chapter 247 of the Laws of Zambia’, have not yielded the desired impact in many areas of the country including Manyinga and Mwansabombwe districts. These two districts continue to report low dog vaccination rates, unrestricted dog movements, and escalating cases of animal and human rabies, along with dog bites. Aligned with global aspirations to achieve zero human rabies cases by 2030, this study scrutinizes the determinants and obstacles hampering the execution of rabies control initiatives in Manyinga and Mwansabombwe. Spanning approximately 11 months, this cross-sectional study gathered pre- and post-vaccination data from 301 households in Manyinga and 100 households in Mwansabombwe. Questionnaires probed knowledge, attitudes, and practices related to rabies prevention and control. A transect survey, key informant interviews, and assessment of rabies vaccination and dog bite records complemented the data collection. Findings revealed that 88% of respondents from both districts possessed knowledge about rabies, confirming affected species and transmission. Moreover, 76.8% in Manyinga and 88.6% in Mwansabombwe were acquainted with rabies prevention and control methods. Concerning dog owners, 89% were aware of rabies, 66% understood its prevention and control, and the majority identified bites as the primary mode of transmission. Vaccination coverage stood at 64% in Manyinga and 21% in Mwansabombwe. Notably, education and occupation exhibited a positive significant association with rabies knowledge. In terms of dog bite cases, Manyinga recorded 538 dog bite cases from 2017 to June 2022 while Mwansabombwe recorded 81 dog bite and 23 jackal bite cases from 2021 to June 2022. The study underscores critical knowledge gaps in rural areas and emphasizes the imperative for enhanced public education and awareness programs, improved rabies surveillance, free mass vaccination campaigns, and community engagement to augment vaccination coverage and knowledge about rabies.

A mathematical model that incorporates human attacks on dogs is presented to explore the dynamics of rabies transmission. The model divided the infection rate into two categories: dog-to-dog transmission rates during the prodromal phase (βdP) and the furious phase (βdF). It has been determined that the model is well-posed and that all of the solutions are positive, as well as the feasibility and positivity of the model's solutions. Both the basic reproduction number (R0) and the effective reproductive number (Re) are computed, and it is demonstrated that the model has a unique disease-free equilibrium that is globally asymptotically stable whenever Re < 1. To identify the model's most sensitive parameters and the ones that intervention efforts should focus on, sensitivity analysis of the effective reproduction number is carried out. We can construct an optimal control system using the state equations, the adjoint equations, and the optimality condition that determines the controls by applying Pontryagin's Maximum/Minimum principle. To help decision-makers in allocating funds for rabies interventions, cost-effectiveness analyses are conducted. To determine the degree to which the intervention strategies are advantageous and cost-effective, this study performs a cost-effective analysis of one or all possible combinations of the optimal rabies control strategies (pre-exposed and post-exposed vaccination for both dog and human populations) for the four different transmission settings. After arranging the techniques to increase effectiveness (total infections prevented), a cost-effective analysis using the Incremental Cost Effectiveness Ratio (ICER) was conducted. The result supports the function that the four intervention options are performing to either completely eradicate or significantly reduce rabies disease among the populations. The dynamical behavior of the system is studied through simulations using the ode45 numerical technique from MATLAB.

Rabies is one of the most significant public and veterinary health problem, causing ap-proximately 59 000 human deaths annually in the developing countries of Asia and Afri-ca. The aetiologic agent, a viral species of the Lyssavirus genus, is highly neurotropic and has a wide host range including terrestrial mammals and several Chiropteran species. The Lyssavirus mokola (MOKV) was first isolated in the late 1960s from organ pools of shrews (Crocidura flavescens manni) in the Mokola forest (Nigeria). To date, at least 30 MOKV isola-tions have been confirmed, all exclusively from Africa, with 73% from southern Africa. There is a limited knowledge about the epidemiology of MOKV and the reservoir host spe-cies is unknown. Here we report on a retrospective identification of a new MOKV from Eswatini, based on molecular characterization of rabies viruses originating from both domestic and African wild cats. A partial region of the lyssavirus genome, encoding the nucleoprotein, was amplified and sequenced. Nucleotide sequence analysis retrospec-tively identified a rare lyssavirus, a Lyssavirus mokola from a domestic cat from Eswatini, whilst the majority of the lyssaviruses were identified as Lyssavirus rabies (98%). Further-more, the nucleotide sequence divergence between the recently identified MOKV isolate, and the historical Lyssavirus mokola isolates ranged from 6.8% and 8.3%. This study further highlights the association between the potential host species of Lyssavirus mokola and the domestic cat as an incidental host. Therefore, continuous vaccination of domestic cats against rabies is crucial even after the elimination of dog-mediated rabies as spill-over re-lated to sylvatic rabies cycles is likely to occur.

Rabies is a life-threatening and vaccine-preventable infectious disease and is triggered by an RNA virus, part of the Rhabdoviridae family, Lyssavirus genus. It is transmitted from infected saliva due to biting, but also from contact with infected blood or organs from rabid animals to humans. Children are at high risk due to their inability to defend themselves from infected animals and most deaths occur with a particularly high incidence in this category (under the age of 15 years). Postexposure prophylaxis (PEP) is standard and life-saving if given early after exposure. We are reporting the case of a ten-year-old female child, who was bitten by an unvaccinated cat ten days before admission to the Paediatric Intensive Care Unit at National Institute for Infectious Diseases „Prof. Dr Matei Bals”, Bucharest. The child had category III cat bites and did not receive PEP. Suggestive clinical manifestations such as an altered general state, psychomotor agitation, auditory and visual hallucinations, and hypersomnia followed by aerophobia and hydrophobia were present. Despite the maximal treatment introduced from early admission (Human Rabies Immunoglobulin, anti-rabies vaccination, orotracheal intubation plus mechanical ventilation, vital functions’ support, parenteral feeding, symptomatic drugs) the evolution of the patient was unfavourable, namely death two months after admission. The diagnosis was established in both pre-mortem by identifying the rabies virus in the cerebrospinal fluid based on reverse transcription-polymerase chain reaction (RT-PCR) and post-mortem by a histopathological examination (which reveals the presence of Babes-Negri bodies in the hippocampus). The objective is to understand the mechanism of infection and mortality from rabies virus in children. Conclusion: The most effective prevention methods are early administration of specific prophylaxis (immunoglobulin and vaccination) and, most importantly, early elimination of animal rabies outbreaks, while rapid diagnosis is helpful in the management of human rabies in humans. The rate of human exposure may be significantly decreased by controlling rabies through pet immunization programs.

Rabies is endemic in southern Bhutan and children were reported to be the most frequent victims. We surveyed the knowledge, attitude, and practices on rabies among school children in three schools located in southern Bhutan. Descriptive statistics and logistic regression were performed to analyze the level of knowledge and variation of perception towards rabies among secondary school children. A total of 701 students (57.9% female, 42.1% male) had participated in the survey of which 98.2% heard about rabies. Most of the students demonstrated a good level of knowledge (59.7%) and a favorable perception towards rabies (57.7%). Multivariable logistic regression analysis revealed the importance of previous rabies awareness campaign increasing the likelihood of positive knowledge about rabies at the individual student and the school level. Similarly, higher grades of students’, employed mothers of the students, and students from villages were associated with more favorable perceptions. Overall, our study in rabies endemic areas of Southern Bhutan showed that most of the students have good knowledge and favorable perception towards rabies. However, we identified several knowledge gaps. Therefore, efforts should be made to address the knowledge gaps through regular awareness programs by actively engaging key stakeholders such as school-teachers and parents.

Background: Rabies, a zoonotic disease caused by the Lyssavirus genus, is widely distributed and a significant cause of human death in Africa and Asia. This study aimed to investigate human rabies exposure and associated risk factors in Northwest Ethiopia. Methods: The study encompassed four zones, seven districts, and four kebeles from each district. The research team developed a questionnaire for data collection. Results: A total of 642 households were interviewed. The present study highlighted an exposure to rabies of 10.47%.”. Over 98% of residents displayed a strong understanding of rabies, with 99.69% correctly identifying rabies as a zoonotic disease. However, 80.22% of the interviewees believed that traditional medicine can treat the disease. Only 21.93% of victims were subjected to post-exposure prophylaxis. More than 87% of exposed residents were affected by the dogs. Of that, 48.70% were bitten by their own dogs. Metema district had the highest risk of exposure (p < 0.001) compared to the other six that were investigated. Dog ownership (p<0.05) and a higher number of households (p<0.01) were also identified as risk factors for rabies exposure. Conclusion: Residence, low education levels, high household density, free-roaming dogs, and dog ownership are key risk factors for rabies exposure. Public health awareness campaigns are crucial to emphasize the risks associated with animal bites.

Estimates of dog population sizes are a prerequisite for delivering effective canine rabies control. However, dog population sizes are generally unknown in most rabies-endemic areas. Several approaches have been used to estimate dog populations but without rigorous evaluation. We compare post-vaccination transects, household surveys and school-based surveys to determine which most precisely estimates dog population sizes. These methods were implemented across 28 districts in southeast Tanzania, in conjunction with mass dog vaccinations, covering a range of settings, livelihoods and religious backgrounds. Transects were the most precise method, revealing highly variable patterns of dog ownership, with human: dog ratios ranging from 12.4:1 to 181.3:1 across districts. Both household and school-based surveys generated imprecise and sometimes inaccurate estimates, possible due to low sample size. Transect data were subsequently used to develop a predictive model for estimating dog populations in districts lacking transect data. We predicted a dog population of 2,316,000 (95% CI 1,573,000-3,122,000) in Tanzania and an average human: dog ratio of 20.7:1. Our modelling approach has the potential be applied to predicting dog population size in other districts where mass dog vaccination is carried out, given census and livelihood data. We recommend transects as a rapid and effective method to refine dog population estimates across large geographic areas and guide dog vaccination programs.

The vaccines have to undergo independent testing at National Control laboratory (NCL) for various quality control tests to ensure safety, purity, and potency before market authorization and surveillance testing. In this study, five different inactivated cell culture rabies vaccine preparations (Aluminum adjuvanted and non-adjuvanted) marketed in India were evaluated using compendial safety test of mouse inoculation test (MIT) for residual live virus. Investigations were done to see the interference of aluminum adjuvant in rabies vaccine preparation to ensure safety as per the in-vivo compendial test for virus inactivation. After intracerebral injections, mice received the aluminum phosphate adjuvanted inactivated rabies vaccine (ARV) had neurological complications that resulted in their deaths between days 4 and 10. Further investigations were carried out to find out the root cause for mice death including fluorescent antibody test (FAT), rabies virus amplification test. Additionally, alamar blue based cell cytoxicity assay was carried out using BHK-21 cells to estimate the cell toxicity of aluminum phosphate adjuvanted inactivated rabies vaccine. The present study highlighted the challenges in ensuring the safety of adjuvated inactivated rabies vaccine at level of NCL. Further, study demonstrated the toxicity of aluminum under both in vitro and in vivo settings, sparking controversy regarding the safety of aluminum adjuvants in vaccines at the suggested compendial limit of 1.25 mg/single human dosage.

Lyme disease is the most common tick-borne illness in the United States, Europe, and Asia. Borrelia burgdorferi, a spirochete bacterium transmitted by the tick vector Ixodes scapularis, causes Lyme disease in the U.S. If untreated, Lyme arthritis, carditis, and meningitis will occur. Given the absence of a human Lyme disease vaccine, we developed a vaccine using the rabies virus vaccine vector, BNSP333, and an outer surface borrelial protein BBI39. To incorporate BBI39 into the RABV virion, we generated a chimeric BBI39 antigen, BBI39RVG, by fusing it with the final amino acids of the RABV glycoprotein. Here we demonstrated that BBI39RVG antigen was incorporated into the RABV virion, and mice vaccinated with our RABV-BBI39RVG vaccine induced high amounts of BBI39-specific antibodies which are maintained long-term. The BBI39 antibodies neutralized Borrelia in vaccinated mice when challenged with Borrelia burgdorferi by either syringe injection or infected ticks, and reduced Lyme disease pathology.

Rabies, caused by the rabies virus (RABV), remains a significant public health issue in the Philippines despite efforts to control it. To eliminate rabies by 2030, effective surveillance strategies are crucial. In this study, we examined RABV evolution and phylodynamics in the Davao Region using genome sequences from Davao City and nearby provinces. We adapted the RABV ARTIC Protocol for Oxford Nanopore High-Throughput Sequencing to optimize workflow efficiency under limited resources. Comparing new virus samples collected from June 2019 to June 2021 (n=38) with baseline samples from June 2018 to May 2019 (n=49), new sub-clades were observed in the phylogenetic tree, suggesting divergence from older variants that were previously undetected. Most of the new viruses belonged to the Asian SEA4_A1.1.1 lineage, but new (SEA4_B1 and SEA4_B1.1) and emerging (SEA4_B1.1_E1) lineages that have never been reported in the Philippines were also identified. The baseline study reported phylogeographic clustering of RABV isolates from the same areas. However, this pattern was disrupted in the current biosurveillance, with variants detected in areas outside the original cluster. Furthermore, our findings revealed significant transmission routes between Davao City and neighboring provinces, contrasting with the predominantly intra-city transmission observed in the baseline study. These results underscore the need for ongoing and timely genomic surveillance to monitor genetic diversity changes and the emergence of novel strains, as well as to track alterations in transmission pathways. Implementing cost-effective next generation sequencing workflows will facilitate the integration of genomic surveillance into rabies control programs, particularly in resource-limited settings. Collaborations between different sectors can empower local laboratories and experts in genomic technologies and analysis.

Since 2012, the northeast of Tunisia has been experiencing an emergence of dog rabies. Published data revealed that the disease is widespread in the north of Tunisia. Even with studies on rabies, lacking knowledge on the associated risk factors was highlighted. Therefore, we conducted a case-control study on dog rabies in northeast Tunisia to identify potential risk factors for dog rabies occurrence. The cases group (n=77) included positive dog rabies confirmed at the referral laboratory using Fluorescent Antibody Test (FAT) in 2014 and 2018. The controls group (n=77) involved all negative cases received at the laboratory and 15-days quarantine dogs that received a certificate of absence of rabies after the observation period. Univariate and multivariate logistic regression was conducted to explore the risk factors (age, sex, breed, confinement, vaccination status, presence of bites, owned or not and origin of dogs, sector, presence of rabies cases, and slaughterhouses, etc.…) related to the occurrence of rabies. The final logistic regression model revealed that unconfined dogs were almost twice as likely to be affected as confined dogs (OR=1.9; 95% CI: 1.17 to 3.27). The odds of rabies occurrence were 25 times higher in areas where rabies cases have been reported between 3 months and 1 year compared to the uninfected areas (25.7; 95% CI: 3.02-219.14). Similarly, the risk of rabies is significantly higher in dogs born in the home to the owner bitch (OR=2.41; 95% CI: 1.14-5.13). Living in areas with rabies cases in the last three months increases by 2.8 times the risk of getting rabies (2.8; 95% CI: 1.16-6.77). However, vaccination reduces by 1.6 times the risk of contracting rabies (0.6; 95% CI: 0.38-0.97). Our findings provide reliable data on dog rabies that will be very useful for decision-makers and could contribute to improving the ongoing national control program of rabies.

Australian bat lyssavirus (ABLV) is a rhabdovirus that circulates in four species of pteropid bats (ABLVp) and the yellow-bellied sheath-tailed bat (ABLVs) in mainland Australia. In the three confirmed human cases of ABLV, rabies illness preceded fatality. As with rabies virus (RABV), post-exposure prophylaxis (PEP) for potential ABLV infections consists of wound cleansing, ad-ministration of the rabies vaccine and injection of rabies immunoglobulin (RIG) proximal to the wound. Despite the efficacy of PEP, the inaccessibility of human RIG (HRIG) in the developing world and the high immunogenicity of equine RIG (ERIG) has led to consideration of human monoclonal antibodies (hmAbs) as a passive immunization option that offers enhanced safety and specificity. Using a recombinant vesicular stomatitis virus (rVSV) expressing the glycoprotein (G) protein of ABLVs and phage display, we identified two hmAbs, A6 and F11, which completely neutralize ABLVs/ABLVp, and RABV at concentrations ranging from 0.19-3.12 µg/mL and 0.39-6.25 µg/mL respectively. A6 and F11 recognize overlapping epitopes in the lyssavirus G protein, ef-fectively neutralizing phylogroup 1 lyssaviruses, while having little effect on phylogroup 2 and non-grouped diverse lyssaviruses. These results suggest A6 and F11 could be effective therapeutic and diagnostic tools for phylogroup 1 lyssavirus infections.

We evaluated the immunogenicity persistence of different immunization regimens of freeze-dried human rabies vaccine (Vero cells) in the 10–60 years age group of the Chinese population. Based on phase III clinical trials of Vero cells, participants who completed the full course of immunization were continuously enrolled in four-dose trial, five-dose, and five-dose control groups. Venous blood samples were collected 12 months after completion of the full immunization for rabies virus-neutralizing antibody (RVNA) testing, and the antibody-positive rate and geometric mean concentration (GMC) were calculated. Analysis of variance was used to compare three groups. The four-dose trial, five-dose trial, and five-dose control groups included 200, 186, and 214 participants, respectively. Twelve months after full immunization, the overall RVNA-positive rates were 97.00%, 93.55%, and 94.86% in the four-dose trial, five-dose control, and five-dose trial groups, respectively. The post-immunization RVNA GMCs values were 2.50, 2.05, and 2.04 IU/mL, respectively, with no statistically significant differences between them. Twelve months after full immunization, the overall RVNA-positive rate remained high, demonstrating satisfactory immunogenicity persistence for both immunization regimens.

Rabies is a fatal zoonosis that is considered a re-emerging infectious disease. Although rabies remains endemic in canines throughout much of the world, vaccination programs have essentially eliminated dog rabies in the Americas and much of Europe. However, despite the goal of eradicating dog rabies in the European Union by 2020, sporadic cases of dog rabies still occur in Eastern Europe, including Georgia. To assess the genetic diversity of strains recently circulating in Georgia, we sequenced 78 RABV-positive samples from brain tissues of rabid dogs and jackals using Illumina short-read sequencing of total RNA shotgun libraries. Seventy-seven RABV genomes were successfully assembled and annotated, 74 of them to the coding complete status. Phylogenetic analyses of the nucleoprotein (N) and attachment glycoprotein (G) genes placed all the assembled genomes into the Cosmopolitan clade, consistent with the Georgian origin of the samples. Amino acid alignment of the G glycoprotein ectodomain identified twelve different sequences for this domain among the samples. Only one of the ectodomain groups contained a residue change in an antigenic site, an R264H change in the G5 antigenic site. Three isolates were cultured, and these were found to be efficiently neutralized by human monoclonal antibody A6. Overall, our data show that recently circulating RABV isolates from Georgian canines are predominantly closely related phylogroup I viruses of the Cosmopolitan clade. Current rabies vaccines should offer protection against infection by Georgian canine RABVs. The genomes have been deposited in GenBank (accessions: OQ603609-OQ603685).

Proteomic analysis of purified rabies virus (RABV) revealed 47 entrapped host proteins within viral particles. Out of these, 11 proteins were highly disordered. Our study was particularly focused on five of the RABV-entrapped mouse proteins with the highest levels of disorder, Neuromodulin, Chmp4b, DnaJB6, Vps37B, and Wasl. We extensively utilized bioinformatics tools, such as FuzDrop, D2P2, UniProt, RIDAO, STRING, AlphaFold, and ELM for comprehensive analysis of the intrinsic disorder propensity of these proteins. Our analysis suggested that these disordered host proteins might play a significant role in facilitating the rabies virus pathogenicity, immune system evasion, and the development of the antiviral drug resistance. Our study highlighted the complex interaction of the virus with its host, with the focus on how the intrinsic disorder can play a crucial role in virus pathogenic processes and suggested that these intrinsically disordered proteins (IDPs) and disorder-related host interactions can also be a potential target for the therapeutic strategies.

Rabies is a neurological disease that causes between 40,000 and 70,000 deaths every year. Once a rabies patient has become symptomatic, there is no effective treatment for the illness, and in unvaccinated individuals, the case-fatality rate of rabies is close to 100%. French scientists Louis Pasteur and Émile Roux developed the first vaccine for rabies in 1885. If administered before the virus reaches the brain, the modern rabies vaccine imparts long-lasting immunity to the virus and saves more than 250,000 people every year. However, the rabies virus can suppress the host’s immune response once it has entered the cells of the brain, making death likely. This study aims to make use of disorder-based proteomics and bioinformatics to determine the impact that intrinsically disordered protein regions (IDPRs) in the proteome of the rabies virus have on the infectivity and lethality of the disease. This study uses the proteome of Rabies Lyssavirus (RABV) strain Pasteur Vaccins (PV), one of the best understood strains due to its use in the first rabies vaccine, as a model. The study suggests that the high levels of intrinsic disorder in the phosphoprotein (P-protein) and nucleoprotein (N-protein) allow them to participate in creation of the Negri bodies and help this virus suppress the antiviral immune response in the host cells. Additionally, the study suggests that there is a link between disorder in the matrix (M) protein and the modulation of viral transcription. The disordered regions in the M protein have a possible role in initiating viral budding within the cell. Furthermore, we checked the prevalence of functional disorder in a set of 37 host proteins directly involved in the interaction with the RABV proteins. The hope is that these new insights will aid in the development of treatments for rabies that are effective after infection.

Most pathogens are capable of infecting multiple host species and such cross-species transmission (CST) can have dramatic consequences, as highlighted by recent disease emergence events affecting human, animal and plant health. Understanding the ecological and evolutionary factors that constrain or facilitate the ability of disease agents to infect and establish in a novel host is therefore a timely and important research area. Previous work across different pathogens, including rabies virus (RABV), found that increased evolutionary distance between hosts reduces the frequency of cross-species transmission and of permanent host shifts. However, whether this effect of host relatedness still holds for transmission among recently diverged hosts, and the importance of this effect relative to other predictors, is not well understood. We addressed this question by quantifying the CST frequency of RABV between North American bat species within the genus Myotis, using a multi-decade data set containing 128 nucleoprotein (N) sequences from ten host species. For comparison, we also conducted an equivalent analysis of a RABV dataset from North American bat species comprising nine genera. We found that at the within genus scale, host relatedness failed to explain the frequency of CST events. However, CST frequency increased with overlap in species’ host range. Moreover, we found evidence of CST occurring among a higher proportion of species, and CST more frequently resulting in sustained transmission in the novel host in the Myotis dataset compared to the multi-genus dataset. Our results suggest that among recently diverged species, the ability to infect a novel host is no longer restricted by physiological barriers but instead is limited by physical contact. Our results improve predictions of where future CST events for RABV might occur and clarify the relationship between host divergence and pathogen emergence.

Effectively preventing and controlling zoonotic diseases requires a One Health approach that involves collaboration across sectors responsible for human health, animal health (both domestic and wildlife), and the environment, as well as other partners. Here we describe the Generalizable One Health Framework (GOHF), a five-step framework that provides structure for using a One Health approach in zoonotic disease programs being implemented at the local, sub-national, national, regional, or international level. Part of the framework is a toolkit that compiles existing resources and presents them following a stepwise schematic, allowing users to identify relevant resources as they are required. Coupled with recommendations for implementing a One Health approach for zoonotic disease prevention and control in technical domains including laboratory, surveillance, preparedness and response, this framework can mobilize One Health and thereby enhance and guide capacity building to combat zoonotic disease threats at the human-animal-environment interface.

A high intensity Catch, Neuter, Vaccinate and Return (CNVR) intervention was used over 5 years to manage the free-roaming dog population of Greater Bangkok, using nearly 300,000 CNVR operations across cover 6 Provinces. An evaluation used multiple methods to assess the impact of this intervention, including clinic data, an observational street survey, an online attitude survey and reported cases of dog rabies confirmed with laboratory testing. The evaluation found evidence of a reduction in free-roaming dog density over time (24.7% reduction over 5 years), a reduction in dog rabies cases (average reduction of 5.7% rabies cases per month) and an improvement in dog-human relationships (a 39% increase per year in free-roaming dogs with visible signs of ownership or care, and a perception of less trouble with free-roaming dogs in districts benefiting from CNVR). The CNVR intervention appears to have been effective at managing the current free-roaming dog population and minimizing one future source of free-roaming dogs by limiting breeding of dogs accessible on the streets. However, there is evidence that other sources of free-roaming dogs exist, presumed to be predominately abandoned or lost owned dogs that were previously inaccessible to the CNVR intervention because they were ordinarily confined or living outside the project area. Hence a fully effective dog population management will require further interventions targeting owned dogs in addition to this CNVR effort.

Bats are reservoirs of many pathogenic viruses including the lyssaviruses rabies virus (RABV) and Australian bat lyssavirus (ABLV). Lyssavirus strains are closely associated with particular host reservoir species, with evidence of specific adaptation. Associated phenotypic changes remain poorly understood but are likely to involve P protein, a key mediator of the intracellular virus-host interface. Here, we examine the phenotype of P protein of ABLV, which circulates as two defined lineages associated with frugivorous and insectivorous bats, providing the opportunity compare proteins of viruses adapted to divergent bat species. We report that key functions of P protein in interferon/STAT1 antagonism and the capacity of P protein to undergo nuclear trafficking differ between lineages. Molecular mapping indicates that these differences are functionally distinct, and appear to involve modulatory effects on regulatory regions or structural impact, rather than changes to defined interaction sequences. This results in partial but significant phenotypic divergence, consistent with ‘fine-tuning’ to host biology, and with potentially distinct properties in the virus-host interface between bat families that represent key zoonotic reservoirs.

The order Mononegavirales harbors numerous viruses of significant relevance for human health, including both established and emerging infections. Currently, vaccines are only available for a small subset of these viruses and antiviral therapies remain limited. Being obligate cellular parasites, viruses must utilize the cellular machinery for their replication and spread. Therefore, targeting cellular pathways used by viruses can provide novel therapeutic approaches. One of the key challenges confronted by both hosts and viruses alike is the successful folding and maturation of proteins. In cells, this task is faced by cellular molecular chaperones, a group of conserved and abundant proteins that oversee protein folding and help maintain protein homeostasis. In this review, we summarize the current knowledge of how the mononegavirales interact with cellular chaperones, highlight key gaps in our knowledge, and discuss the potential of chaperone inhibitors as antivirals.

Engineered viral vectors designed to deliver genetic material to specific targets offer significant potential for disease treatment, safer vaccine development, and the creation of novel biochemical research tools. Viral tropism, the specificity of a virus for infecting a particular host, is often modified in recombinant viruses to achieve precise delivery, minimize off-target effects, enhance transduction efficiency, and improve safety. Key factors influencing tropism include surface protein interactions between the virus and host cell, the availability of host cell machinery for viral replication, and the host immune response. This review explores current strategies for modifying the tropism of recombinant viruses by altering their surface proteins. We provide an overview of recent advancements in targeting non-enveloped viruses (Adenovirus and Adeno-associated vi-rus) and enveloped viruses (Retro/Lentivirus, Rabies, Vesicular stomatitis virus, and Herpesvirus) to specific cell types. Additionally, we discuss approaches such as rational design, directed evo-lution, in silico and machine learning-based methods for generating novel AAV variants with desired tropism, and the use of chimeric envelope proteins for pseudotyping enveloped viruses. Finally, we highlight the applications of these advancements, and discuss the challenges and future directions in engineering viral tropism.

Despite being a rare disease worldwide, rabies has the highest morbidity and mortality rates, with nearly all symptomatic cases leading to coma and death. Rabies represents an infectious disease caused by the Rabies virus (RABV), which is part of the Lyssavirus group and the Rhabdoviridae family, and it mainly spreads through the bite and scratch of an infected mammal, but particularly of wild animals, such as bats, foxes, wolves and racoons, and of domestic animals, such as dogs and cats, in rabies-prone areas of the world. Airborne transmission has been deemed as extremely rare, and no clinical case as such has been recorded worldwide yet, except in the enclosed environment, such as research laboratories and caves where infected bats are present. Domestic mammals, such as dogs and ferrets, represent other important reservoirs of disease transmission, and the human cases of Asia and Africa amount approximately 95% of all human cases worldwide. Infected animals most commonly start transmitting the virus once the first symptoms have occurred, and if they experience disease aggravation and death within 10 days, a case of rabies is registered, more easily if the incidence occurred in the urban area and then, any person or animal that had been potentially exposed are strongly recommended to receive the inoculation. It is rare for asymptomatic mammals to transmit the illness. Most First-World and several Second-World countries have recently been declared dog rabies-free by the World Health Organization. The disease can only be treated prophylactically, with three doses of a vaccine containing an inactivated form of RABV, or with five doses of the vaccine and two doses of anti-RABV immunoglobulins within 28 days if the patient is believed to have been exposed to the virus beforehand. It has been projected that, once the viral load reaches elements of the central nervous system, prophylactic approaches are no longer effective, even if symptoms have not begun yet, and this highlights the urgent trait of the medical condition, strongly recommending exposed people to receive the prophylactic doses immediately after the potential exposure to the virus. The pathogen first infects the bodily fluids, before reaching the peripheral nervous system, from where it will gradually move toward the spinal cord or the encephalon, at a speed of movement ranging from 1 to 40 cm per day. It was also found, in extremely rare circumstances, to infect the nasopharyngeal cavity and the lungs. The primary cause of a successful, gradual advance of the viral load toward the point of clinical no-return for the patient - the CNS - is a complex mechanism of induced innate immune evasion, with the interferon system being heavily targeted and silenced by RABV proteins. The ‘Milwaukee’ protocol is locally believed to decrease the mortality rate of the clinical illness to approximately 80%, although significantly more research is required in this sense. First-line immune evasion represents the central mechanism developed by viruses during their evolutionary process to gain control over human immunity, so it could be the development and adjustment of a counter-offensive to this evolutionary operating system that could address the core elements of the problem. Human recombinant Type I and Type III Interferons were found to be significant vaccine adjuvants and to considerably delay the clinical onset of the disease. Despite their central role in natural immunity-based prophylaxis, vaccine support and, in often cases, vaccination per se, a local administration of IFNs as such may not be enough to tackle the core problem of the endemic disease, and a specific and systemic treatment of potential host cells with IFN I and III, as well as IFN-stimulating proteins, may constitute a major research requirement in the coming years of disease investigation, as the inoculation efforts with the inactivated virus and immunoglobulin administration continue. The administration of a relatively low dosage of somatic Natural Killer cells, gamma-interferon and perhaps, of somatic helper CD4+ and somatic cytotoxic CD8+ T-lymphocytes treated with alpha-, beta- and lambda-interferon could be merged with the administration of a similar dosage of alpha-, beta- and lambda-interferon during the efforts to develop an effective and less costly prophylactic vaccine against rabies. A combination of a nasal substance containing a low dosage of IFN I and III with a reduced concentration of neutralized RABV copies, and/or with a low dose of anti-RABV IgA antibodies, could also be tested for humans for the purposes of pre- and post-exposure prophylaxis.