Search Results (729)

The rabies virus (RABV) phosphoprotein (P protein) has multiple functions, including acting as the essential non-catalytic cofactor of the viral polymerase (L protein) for genome replication and transcription; the principal viral antagonist of the interferon (IFN)-mediated innate immune response; and the chaperone for the viral nucleoprotein (N protein). Although P protein is known to undergo phosphorylation by cellular kinases, the location and functions of the phosphorylation sites remains poorly defined. Here, we report the identification by mass-spectrometry (MS) of residues of P protein that are modified by phosphorylation in mammalian cells, including several novel sites. Analysis of P protein with phospho-mimetic and phospho-inhibitory mutations of three novel residues/clusters that were commonly identified by MS (Ser48, Ser183/187, Ser217/219/220) indicate that phosphorylation at each of these sites does not have a major influence on nuclear trafficking or antagonistic functions toward IFN signalling pathways. However, phosphorylation of Ser48 in the N-terminus of P protein impaired function in transcription/replication and in the formation of replication structures that contain complexes of P and N proteins, suggestive of altered interactions of these proteins. The crystal structure of P protein containing the S48E phospho-mimetic mutation indicates that Ser48 phosphorylation facilitates the binding of residues 41–52 of P protein into the RNA-binding groove of non-RNA-bound N protein (N⁰), primarily through the formation of a salt bridge with Arg434 of N protein. These data indicate that Ser48 modification regulates the cycling of P-N⁰ chaperone complexes that deliver N protein to RNA to enable transcription/replication, such that enhanced interaction due to S48E phospho-mimetic mutation reduces N protein delivery to the RNA, inhibiting subsequent transcription/replication processes. These data are, to our knowledge, the first to implicate phosphorylation of RABV P protein in conserved replication functions of the P gene. Full article

Chiroptera includes over 1400 bat species, with at least 35 of these present in Italy. Due to their role as Lyssavirus reservoirs, bats found dead, with and without signs suggestive of this infection, are routinely submitted to the laboratory network of the Istituti Zooprofilattici Sperimentali in the framework of the rabies national passive and active surveillance program. Carcasses and biological samples collected from January to December 2021 in Latium and Tuscany, regions of our jurisdiction, were further screened for the presence of Coronaviruses (CoVs) and Herpesviruses using pan-family virus PCR tests, and relative PCR products were Sanger sequenced. Genetic characterization through sequencing detected AlphaCoVs in Miniopterus schreibersii and Beta- and Gammaherpesviruses in Tadarida teniotis. Samples were also submitted to bat genetic species identification. Full article

The history of the rabies virus dates back four millennia, with the virus being considered by many to be the first known transmitted between animals and humans. In Brazil, rabies virus variants associated with terrestrial wild animals, marmosets, and different bat species have been identified. In this study, bat samples from different regions of São Paulo State, in Southeast Brazil, were analyzed to identify their genetic variability and patterns. A total of 51 samples were collected over ten years (1999–2009) and submitted to the immunofluorescent technique using monoclonal antibodies for antigenic profile detection (the diagnostic routine used in Latin American countries) and genetic evolution analysis through maximum likelihood approaches. Three antigenic profiles were detected: one related to the rabies virus maintained by hematophagous bat populations (AgV3), part of the monoclonal antibody panel used, and two other profiles not included in the panel (called NC1 and NC2). These antigenic profiles were genetically distributed in five groups. Group I was related to hematophagous bats (AgV3), Groups II and III were related to insectivorous bats (NC1) and Groups IV and V were also related to insectivorous bats (NC2). The results presented herein show that genetic lineages previously restricted to the northwest region of São Paulo State are now found in other state regions, highlighting the need for a comprehensive genetic study of bat rabies covering geographic and temporal space, through expanded genomic analysis using a standard genomic fragment. Full article

Nitrogen vacancy (NV) diamonds are promising room temperature quantum sensors. As the technology moves towards application, efficient use of energy and cost become critical for miniaturization. This work focuses on microwave-based spin control using the short-circuited end of a slot line, analyzed by finite element method (FEM) for magnetic field amplitude and uniformity. A microstrip-to-slot-line converter with a $-10$ dB bandwidth of $3.2$ GHz was implemented. Rabi oscillation measurements with an NV microdiamond on a glass fiber show uniform excitation over $1.5$ MHz across the slot, allowing spin manipulation within the coherence time of the NV center. Full article

The number of puppies imported to the United Kingdom (UK) from the European Union (EU) is increasing, as are concerns about the legality, welfare and potential disease risks associated with their trade. This study aimed to use an online survey to collect questionnaire data from UK residents about their awareness and knowledge of EU Pet Passports, illegal puppy importation, and dog-borne exotic diseases and the risks they might pose to UK-resident dogs and people. Valid responses were collected from n = 7184 UK residents, mainly current dog owners. We identified substantial knowledge gaps relating to EU Pet Passports, how puppies might be moved across Europe, and the legal age of importation. Awareness of exotic diseases was poor; knowledge about the risks they posed was even poorer, particularly in relation to dog–human transmission. With some respondents not even being confident that they had heard of rabies, there is substantial work to do to ensure that the UK canine and human populations remain safe in the face of significant welfare and exotic disease threats. These knowledge deficits may facilitate the burgeoning illegal trade in puppies across the EU. Legal change may be required to prevent consumers from unintentionally supporting poor welfare sources of puppies. Full article

Rabies is a fatal but entirely vaccine-preventable disease, with the highest risk in areas where free-roaming domestic dogs are prevalent. Understanding dog owners’ knowledge, attitudes, and practices (KAP) is crucial for shaping effective rabies control strategies. This scoping review aimed to synthesize global evidence from studies evaluating dog owners’ KAP to identify behavioral factors relevant to rabies prevention and control. A systematic literature search was conducted using PubMed, Web of Science, and Scopus, covering the period from 2012 to 2025. Seventy full-text articles were included based on predefined criteria. The findings reveal substantial gaps in dog owners’ knowledge, beliefs, and behaviors regarding rabies prevention. While general awareness of rabies is high among dog owners, their knowledge about transmission, clinical signs, and the fatal nature of the disease is inconsistent, with significant variability across studies. The vaccination uptake also varied widely across studies, ranging from less than 1% to over 90%, with no study reporting full coverage. Furthermore, a strong positive correlation was found between vaccination practice and the awareness of vaccine benefits (r = 0.69, p = 0.004). Common barriers to vaccination include lack of information, vaccine accessibility, distance to clinics, and personal constraints. These insights underscore the importance of early and targeted communication about vaccination campaigns. Future research should focus on periodically evaluating KAP before and after interventions to better inform rabies control efforts. Full article

Background: Since the introduction of Pasteur’s rabies vaccine in 1885, rabies prophylaxis and post-exposure prophylaxis (PEP) have been widely administered globally under the recommendation of the World Health Organization (WHO). However, 124 documented cases of PEP failure had been reported worldwide between 1980 and 2023. Additionally, sporadic media reports from China showed occasional PEP failures between 2017 and 2024. Rabies remains a serious public health problem in over 150 countries and regions. Methods: In this review, we summarize PEP procedures recommended by the Advisory Committee on Immunization Practices (ACIP) and the WHO. We also analyze potential contributing factors to PEP failure, propose a concept of circulating antibodies, and discuss their roles in PEP. Furthermore, we summarize key guidelines for clinical trial design from the U.S. Food and Drug Administration (FDA) and China’s Center for Drug Evaluation (CDE), as well as the latest developments in monoclonal antibody (cocktail) therapies. Results: Adherence to core PEP practices, such as wound cleansing, infiltration of wounds with immunoglobulin (mAbs), and administration of vaccines, and broader societal involvement are crucial for preventing rabies infection in most cases. For high-risk exposures or immunocompromised individuals, the provision of circulating antibodies through high-dose human rabies immune globulin (HRIG) or mAbs is of utmost importance for preventing PEP failure. Conclusions: Early, high-concentration circulating antibodies are important for preventing PEP failure. Addressing the global issue of rabies requires involvement of the entire society. Only through collective efforts can we tackle this neglected disease and achieve WHO’s goal of “zero by 30”. Full article

Urbanization is causing a decrease in agricultural land. This leads to changes in cropping patterns, irrigation water availability, and water allowance. Therefore, change in cropping pattern, irrigation water availability, and water allowance were investigated in the Multan region of Pakistan using remote sensing and GIS techniques. The multi-temporal Landsat images with 30 m resolution were acquired for both Rabi (winter) and Kharif (summer) seasons for the years of 1988, 1999 and 2020. The image processing tasks including layer stacking, sub-setting, land use/land cover (LULC) classification, and accuracy assessment were performed using ERDAS Imagine (2015) software. The LULC maps showed a considerable shift of orchard area to urban settlements and other crops. About 82% of orchard areas have shifted to urban settlements and other crops from 1988 to 2020. The LULC maps for Kharif season indicated that cropped areas for cotton have decreased by 42.5% and the cropped areas for rice have increased by 718% in the last 32 years (1988–2020). During the rabi season, the cropped areas for wheat (Triticum aestivum L.) have increased by 27% from 1988 to 2020. The irrigation water availability and water allowance have increased up to 125 and 110% due to decrease in agricultural land, respectively. The overall average accuracies were found as 87 and 89% for Rabi and Kharif crops, respectively. The LULC mapping technique may be used to develop a decision support system for evaluating the changes in cropping pattern and their impacts on net water availability and water allowances. Full article

Southeast Asia is a biodiversity hotspot for bats that can carry lyssaviruses, causing zoonotic diseases. This study detects and quantifies IgG antibodies against Lyssavirus glycoproteins in cave-dwelling bat populations on Cat Ba Island, northern Vietnam, to determine their past exposure history and the prevalence of immune responses. Samples were collected from five caves, encompassing three families and five key species (Hipposideros armiger, H. alongensis, H. poutensis, Taphozous melanopogon, and Myotis pilosus). Using ELISA with the Platelia™ Rabies II kit,(Bio-Rad Laboratories, Marnes-la-Coquette, France) 29.0% (18/62) of the bats tested positive, indicating prior exposure. The detection rate was slightly higher in females (35.7%) than in males (30.4%). Lyssavirus-specific antibodies were detected in four species, with the highest levels found in M. pilosus, followed by H. alongensis, H. armiger, and H. poutensis; no positives were found in T. melanopogon samples. One bat exhibited high seroconversion value (>4 EU/mL). The findings provide serological evidence of widespread lyssaviruses exposure in asymptomatic bats on Cat Ba Island, confirming their role as reservoirs that elicit an immune response without exhibiting rabies symptoms. This highlights the role of caves in facilitating close contact among bats, which may increase viral transmission, highlighting the need for continued surveillance in these unique roosting environments. Full article

Background: Many new mRNA-based vaccine candidates in liquid mRNA-LNP formulations are under development; however, their stability limitations necessitate frozen storage, posing a significant challenge for long-term storage and transportation. Methods: In this study, an mRNA-LNP rabies vaccine, ABO1005, was prepared, freeze-dried and stored at 2–8 °C for 12-month storage stability evaluation. The immunogenicity, vaccine potency (the NIH method), and protective efficacy of ABO1005 were assessed in mice or dogs and compared to a commercialized inactivated vaccine. Results: Research conducted in mice indicated that the lyophilized vaccine exhibited comparable immunogenicity to its liquid form counterpart. Furthermore, the vaccine candidate elicited a robust humoral response lasting at least 175 days, and the specific antibody titers were not affected by the pre-administration of hyperimmune serum. In comparison to the commercialized inactivated vaccine (HDCV or PVRV), ABO1005 elicited significantly higher levels of humoral and cellular immunity. Vaccine potency testing (NIH) revealed that the potency of ABO1005 at 15 μg/dose was 8.85 IU/dose, which is substantially higher than the standard required for the lot release of rabies vaccines for current human use. In a post-exposure prophylaxis (PEP) study in Beagle dogs, the lyophilized vaccine provided 100% protection for dogs following a two-dose regimen (D0-D7), whereas commercially approved inactivated vaccine offered 83% protection. After storage at 2–8 °C for 12 months, no notable changes were observed in the particle size, encapsulation efficiency, and integrity of mRNA or in the immunogenicity of the lyophilized vaccine. Conclusions: This study successfully developed a formulation and process of freeze-drying for a rabies mRNA vaccine, paving the way for future lyophilized mRNA vaccine development. Full article

In Tunisia, rabies is endemic and represents a significant public health issue. The objectives of our study were to describe the epidemiological and clinical profiles of human rabies cases and report the risk factors associated with their occurrence. We conducted a retrospective, descriptive, and analytical study of human rabies cases confirmed at the Rabies Laboratory of the Pasteur Institute in Tunis from January 2000 to November 2022. Temporal–spatial, sociodemographic, and clinical variables and factors related to the exposure context, post-exposure, and response were collected for each patient. A total of 58 human rabies cases were identified. The governorates of Kairouan and Nabeul were the most affected, with a predominance of rural areas (77%, 34/44). The highest number of cases was recorded between May and November (74%, 43/58). The cases predominantly involved males, with the most affected age group being individuals aged from 31 to 59 years (30%, 17/57). Rabies transmission was primarily due to dogs (86%, 43/50) and a single bite (55%, 32/58). After an average incubation period of 60.3 days, hydrophobia and behavioral disturbances were the most common symptoms. This study demonstrates that the risk of human rabies remains present in Tunisia, highlighting the need to improve awareness and post-exposure prophylaxis practices. Full article

Zoonotic diseases are transmitted from animals to humans, and they impose a significant global burden by impacting both animal and human health. It can lead to substantial economic losses and cause millions of human deaths. The emergence and re-emergence of zoonotic diseases are heavily influenced by both anthropogenic and natural drivers such as climate change, rapid urbanization, and widespread travel. Over time, the unprecedented rise of new and re-emerging zoonotic diseases has prompted the need for rapid and effective vaccine development. Following the success of the COVID-19 mRNA vaccines, mRNA-based platforms hold great promise due to their rapid design, swift development and ability to elicit robust immune responses, thereby highlighting their potential in combating emerging and pre-pandemic zoonotic viruses. In recent years, several mRNA vaccines targeting emerging and re-emerging zoonotic viral diseases, such as rabies, Nipah, Zika, and influenza, have advanced to clinical trials, demonstrating promising immunogenicity. This review explores recent advances, challenges, and future directions in developing mRNA vaccines against emerging and re-emerging zoonotic viral diseases. Full article

Rabies, a viral encephalitis caused by rabies virus (RABV), is 100% fatal upon the onset of symptoms. Effective post-exposure prophylaxis (PEP) measures are available, but they are often difficult to access in low-income countries. WHO estimates about 59,000 deaths due to rabies globally, and the majority are contributed by developing countries. Hence, developing drugs for the treatment of post-symptomatic rabies is an urgent and unmet demand. It is worth noting that previous efforts regarding antiviral strategies, such as small-interfering RNA, antibodies and small-molecule inhibitors, against the rabies virus have failed to show efficacy in pre-clinical studies, especially when the virus has reached the central nervous system (CNS). Therefore, drug repurposing seems to be an alternative tool for the development of new anti-rabies drugs. We validated and used a high-throughput, FITC-conjugated antibody-based flow cytometry assay to expedite the identification of repurposable new drug candidates against the RABV. The assay was validated using ribavirin and salinomycin as reference compounds, which showed EC50 values of 10.08 µM and 0.07 µM, respectively. We screened a SelleckChem library comprising 3035 FDA-approved compounds against RABV (CVS-11) at 10 µM concentration. Five compounds (clofazimine, tiamulin, difloxacin, harringtonine and homoharringtonine) were active against RABV, with greater than 90% inhibition. Homoharringtonine (HHT) identified in the present study is active against laboratory-adapted RABV (CVS-11) and clinical isolates of RABV, with an average EC50 of 0.3 µM in both BHK-21 and Neuro-2a cell lines and exhibits post-entry inhibition. Full article

Free-roaming dogs (FRDs) pose significant public health and animal welfare challenges globally, particularly in developing nations, where they serve as major reservoirs for zoonotic diseases, especially rabies. Determining the population size of FRDs, their demographic characteristics, and the factors influencing population size is essential for effective dog population management, which constitutes a critical component of rabies control programs. This study aimed to estimate the size and describe the characteristics of the FRD population and evaluate temporal and spatial variations in FRD sightings on the Kasetsart University Kamphaeng Saen Campus, Thailand. Factors influencing population size were assessed using a negative binomial regression model. The campus area was divided into 52 accessible blocks for dog-counting purposes. In 2018 and 2019, each block was surveyed once annually for a comprehensive census. In 2021, each block was surveyed on six occasions, three in the morning and three in the evening, using a photographic recapture method. Dogs sighted were photographed, and their characteristics and spatial coordinates were recorded. The total population sizes of FRDs on campus were documented as 529, 577, and 531 in 2018, 2019, and 2021, respectively. Intact female dogs predominantly constituted the majority of the FRD population. Regression analysis indicated a significant association between the number of FRDs within a block and the number of intact adult female dogs. The percentage of dogs sighted per occasion was significantly higher during evening surveys compared to morning surveys. This study proposes a practical census approach that combines total block counts with photographic recapture to estimate the population of FRDs. The findings offer valuable insights for developing effective dog population management strategies and planning rabies vaccination campaigns. Full article

Zoonotic diseases are a persistent public health concern, especially in low- and middle-income countries like Brazil. This cross-sectional study evaluated the knowledge and perceptions of 132 high school students (70 public and 62 private) in Goiânia, Brazil, regarding zoonoses, using a structured questionnaire. Statistical analyses (Chi-square and Fisher’s exact tests) revealed significant differences (p < 0.05) between public and private school students in knowledge levels, pet care practices, and the awareness of zoonotic risks. While pet ownership was common in both groups, only 53% of private and 21% of public school students correctly defined “zoonosis.” Rabies, taeniasis, leptospirosis, tuberculosis, cysticercosis, cutaneous larva migrans, and leishmaniasis were the most frequently cited diseases, with private school students demonstrating greater recognition across all categories. However, most participants lacked detailed knowledge about transmission routes and prevention. Misconceptions—such as zoonoses affecting only low-income populations—were also identified. Preventive actions like sanitation, public education, and vaccination were commonly suggested but not consistently linked to zoonoses. These findings highlight critical educational gaps and emphasize the need to incorporate One Health principles into school curricula to improve youth understanding and support public health strategies for zoonosis prevention. Full article