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Canine distemper (CD) is a deadly, multi-system infection caused by a Morbillivirus. The canine distemper virus (CDV) frequently affects the nervous system with demyelinating leukoencephalitis, the most common neurological lesion. The disease has been linked to multiple sclerosis (MS) in humans due to similar clinical presentation and pathophysiological mechanisms. In MS, visual evoked potentials (VEPs) have been identified as a reliable marker for disease progression, enabling the early detection of clinically suspected lesions. The aim of this study was to determine if there are any abnormalities in VEP responses in dogs with neurological CD. Visual evoked potentials and electroretinogram (ERG) were recorded at both the cranial and spinal levels in dogs naturally infected with CDV and in healthy dogs. The results in the CDV-infected group revealed a bilateral increase in the latency of N1, P1, N2, P2, and N3 waves of the VEPs, without any alterations in their amplitudes. No significant differences were observed in the ERG between the groups. These results suggest that altered VEP responses could serve as an early diagnostic indicator of neurological damage caused by distemper. Therefore, conducting these studies could potentially aid in the detection of central nervous conduction disorders during the subclinical phases of the disease. [ABSTRACT FROM AUTHOR]

Canine distemper virus (CDV) poses a substantial threat to diverse carnivorans, leading to systemic and often fatal diseases. Accurate and prompt diagnosis is paramount for effective management and curbing further transmission. This study evaluates the diagnostic performance of droplet digital PCR (ddPCR) in comparison to conventional reverse-transcription (RT-PCR) and quantitative reverse-transcription real-time PCR (RT-qPCR). Seventy-six clinical samples were collected from dogs with CDV symptoms diagnosed by specialized veterinarians, and sixteen samples from apparently healthy individuals. Conventional PCR, quantitative real-time PCR, and ddPCR were deployed, and their diagnostic capabilities were meticulously assessed. DdPCR exhibited heightened analytical sensitivity, reaching a detection limit of 3 copies/μL, whereas RT-qPCR had a detection limit of 86 copies/μL. The comparative analysis between clinical diagnosis and molecular techniques, including RT-PCR and RT-qPCR, demonstrated low concordance, with Kappa coefficients of 0.268 and 0.324, respectively. In contrast, ddPCR showed a moderate concordance, with a Kappa coefficient of 0.477. The sensitivity was 42.4% for RT-PCR, 57.9% for RT-qPCR, and 72.4% for ddPCR, with 100% specificity for all methods. This study underscores ddPCR's superior sensitivity and agreement with clinical CDV diagnosis, even at low viral concentrations, suggesting it as a promising alternative for CDV diagnosis. [ABSTRACT FROM AUTHOR]

We describe a natural coinfection with canine distemper virus (CDV) and yellow fever virus in a free-ranging neotropical primate of the genus Callithrix, found dead in the northeastern region of Brazil. The laboratory diagnosis included histopathology, immunohistochemistry, rRT-PCR, and phylogenetic analyses. The CDV sequences from this primate in Brazil represent a divergent lineage in Rio Grande do Norte, closely related to genotypes EU1/South America 1 and South America 2. To our knowledge, this is the first report of natural coinfection by CDV and yellow fever virus in a neotropical primate, underscoring the need to further investigate the circulation of this virus in Brazilian nonhuman primates and its potential implications for wildlife conservation. [ABSTRACT FROM AUTHOR]

Unlike most canids, versatile and capable of navigating vast landscapes, endangered Ethiopian wolves (Canis simensis) are endemic to an archipelago of Afroalpine islands. As a habitat specialist, the Ethiopian wolf is ill‐equipped to move across a highly transformed and densely populated agriculture matrix. Hard borders imposed by expanding subsistence agriculture lock Ethiopian wolves into further isolation, with few opportunities for dispersal and recolonisation. We report and evaluate empirical information from long‐term monitoring across the species' range to understand processes of habitat loss, recolonisation and extinction in recent and historical times, and to assess what conservation measures and strategies would ensure their persistence. Ethiopian wolves occurred in six isolated populations, totalling 454 wolves (population sizes ranged between 281 and 24) occupying 2700 km2 of Afroalpine habitat. We describe three population extinctions and three local extinctions within fragmented populations, and present evidence of factors accelerating the extinction process, such as disease (rabies and canine distemper virus), persecution, road kills and poisoning. Of all the suitable habitat available to wolves, 86% was included within nine protected areas, including three new Community Conservation Areas and two national park extensions in the past 10 years. As all Ethiopian wolf populations are small and vulnerable to stochastic events and environmental perturbation, conservation efforts to ensure the long‐term survival of the species need to integrate: 1) support for protected areas to halt agriculture encroachment and to regulate sustainable uses of natural resources; 2) efforts to minimise all causes of mortality, including but not limited to disease; and 3) conservation translocations to overcome fundamental barriers to dispersal in the highlands of Ethiopia. [ABSTRACT FROM AUTHOR]

Environmental changes associated with global warming create new opportunities for pathogen and parasite transmission in Arctic wildlife. As an apex predator ranging over large, remote areas, changes in pathogens and parasites in polar bears are a useful indicator of changing transmission dynamics in Arctic ecosystems. We examined prevalence and risk factors associated with exposure to parasites and viral and bacterial pathogens in Chukchi Sea polar bears. Serum antibodies to six pathogens were detected and prevalence increased between 1987–1994 and 2008–2017 for five: Toxoplasma gondii, Neospora caninum, Francisella tularensis, Brucella abortus/suis, and canine distemper virus. Although bears have increased summer land use, this behavior was not associated with increased exposure. Higher prevalence of F. tularensis, Coxiella burnetii, and B. abortus/suis antibodies in females compared to males, however, could be associated with terrestrial denning. Exposure was related to diet for several pathogens indicating increased exposure in the food web. Elevated white blood cell counts suggest a possible immune response to some pathogens. Given that polar bears face multiple stressors in association with climate change and are a subsistence food, further work is warranted to screen for signs of disease. [ABSTRACT FROM AUTHOR]

Background and Aim: Viral gastroenteritis in canines is primarily caused by the canine parvovirus 2 (CPV-2). Infections by this virus can cause severe consequences in dogs, such as fever, vomiting, diarrhea, septicemia, systemic inflammation, and immunosuppression. Therefore, the mortality rate of persistent infections caused by this virus is significantly high. The capsid protein VP2 genome of canine parvovirus has undergone many changes, resulting in the emergence of different genotypes, including CPV-2a, CPV-2b, and CPV-2c. Diagnostic procedures often lack the necessary specificity for early infection diagnosis. Early detection of the infection enhances the likelihood of canine survival because the canine will receive prompt therapy. Hence, this study aimed to develop a quantitative polymerase chain reaction (qPCR)-based diagnostic technique using SYBR Green for the rapid and accurate detection and quantification of CPV-2. Materials and Methods: The assay was specifically designed to identify a portion of the conserved NS gene using primers that amplify a 125-bp fragment. The qPCR method was executed in the fast mode to expedite the process using Power up SYBR Green Master Mix reagent. A standard curve was constructed using the amplified and purified PCR product of the NS gene. Results: The limit of detection and quantification were determined in the one amplified-DNA copy. The standard curve showed an efficiency of 99.5% and inter- and intra-assay coefficients of variation of 0.387%-0.976% and 0.085%-0.430%, respectively. The assay was specific for the amplification of CPV-2, as no amplification was observed for other viral genomes (canine adenovirus II, canine distemper virus, canine coronavirus, and canine astrovirus) or from the negative controls. Inter- and intra-tests for repeatability showed low test variability around the run time. To validate the present assay, 200 samples of fezzes from canines with gastroenteritis and symptoms associated with enteric infection were tested using the qPCR protocol. From the analyzed samples, 136 were positive for CPV-2 by qPCR assay, of which 110 were before diagnostic positive for the virus by endpoint PCR, showing high sensitivity of the current assay. CPV-2 was detected in dogs over 2 weeks old up to dogs 9 years old, where the highest viral concentration found was 16429595 gene copies in dogs aged 2 weeks. Conclusion: In the present study, a rapid, specific, repeatable, and sensitive assay was developed for the detection and quantification of CPV-2. Furthermore, it was demonstrated that in the population of domestic dogs in Ecuador affected with gastrointestinal disease, the virus is presented in dogs of different ages and not only in young dogs. [ABSTRACT FROM AUTHOR]

Canine Distemper Virus (CDV) infection causes a multifocal demyelinating progressive disease within the central nervous system (CNS) that results in wide range of neurological symptoms. Pathological changes in the brain or CNS could be observed by analyzing cerebrospinal fluid (CSF). Therefore, this study aimed to investigate the concentrations of amino acids in the CSF of dogs with neurological form of CDV infection, and to identify amino acids as markers that can be used in the diagnosis, pathogenesis, and treatment of the disease. Heathy dogs (n=6), confirmed by clinical and laboratory examinations (Healthy group), and CDV-infected dogs (n=10) with neurological symptoms, confirmed by clinical, laboratory, and rapid diagnostic test kits (Distemper group), were used. CSF samples were obtained with an appropriate method and were subjected to 1H-NMR analysis. Identification was made on 10, whereas quantification on 8 amino acids. L-tyrosine, L-phenylalanine, L-threonine, and L-alanine concentrations were significantly lower, while L-histidine and L-tryptophan were significantly higher than the Healthy group (p<0.05). It was concluded that L-tyrosine could be used for assessing mental status changes, L-phenylalanine for evaluating neuroprotective responses, L-threonine and L-histidine for gauging the extent of neurodegeneration and ventricular degeneration, L-alanine for exploring cellular stress and energy metabolism, and L-tryptophan for understanding the process of sympathetic nervous system activation. [ABSTRACT FROM AUTHOR]

Canine distemper virus (CDV) poses a significant threat to dogs and wildlife worldwide, and this study sought to provide an updated genetic characterization of CDV field strains in Chile during 2022–2023. We collected samples from 52 suspected CDV cases in domestic dogs and detected viral RNA through real‐time RT‐PCR in 28 dogs (54%). Factors like age and vaccination status were determinants of CDV positivity, with young adult and unvaccinated dogs facing a higher infection risk. We isolated CDV from positive samples in VeroDogSLAM cells. From these isolates and direct samples, we obtained sequences and estimated the phylogeny based on gene H. CDV isolates from nasal and conjunctival swabs exhibited cytopathic effects, and sequence analysis unveiled a substantial genetic diversity among the strains. Chilean CDV strains demonstrated a genetic distance to vaccine strains of approximately 10%, antigenic‐change‐related amino acid substitutions, and novel putative glycosylation sites. In the phylogeny, Chilean CDV field strains clustered into two lineages, Europe/South America‐1 and North/South America‐4, indicating the emergence of the North/South America‐4 lineage in Chile and underscoring the genetic complexity of CDV in the country. Interestingly, certain Chilean viruses shared a close common ancestor with Brazilian and Peruvian viruses, suggesting viral spreading patterns. Further investigations are warranted to comprehend the potential antigenic implications of these genetically diverse CDV strains. [ABSTRACT FROM AUTHOR]

Background: Morbillivirus canis is the etiological agent of a highly contagious disease that affects diverse domestic and wild animals. Vaccination is considered the most suitable strategy for controlling CDV dissemination, transmission, and distemper disease. However, the emergence of new CDV strains has led to the need to update the current vaccine strategies employed to prevent CDV infection in domestic and wild animals. Currently, there is a lack of effective alternatives for wild animals. Diverse computational tools, especially peptide-based therapies, enable the development of new universal vaccines. Objective: The aim of this study was to evaluate the safety and humoral and cellular immune response of a new generation of vaccines based on CDV peptides as single-peptide mixtures or multiepitope CDV polypeptides in mice. Methods: Twenty-four BALB/c mice were subjected to a three-dose regimen for 28 days. Seroconversion was evaluated via ELISA, and cellular immune responses were evaluated via flow cytometry through activation-induced markers (AIMs). Results: Compared with the placebo, the peptide mixture and multiepitope CDV polypeptide were safe, and seroconversion was statistically significant in the multiepitope CDV polypeptide and commercial vaccine (CV) groups. The numbers of antigen-specific CD4+CD134+ and IFN-γ+ T cells, CD8+ T cells and TNF-α- and IL-6-producing cells were greater in the mice immunized with the multiepitope CDV polypeptide than in the control mice. Conclusion: This combined approach represents a potential step forward in developing new immunization candidates or enhancing current commercial vaccines to control CDV disease in domestic dogs and wild animals. [ABSTRACT FROM AUTHOR]

Abstract Canine distemper virus (CDV) is a highly contagious and often fatal disease affecting wild and domesticated carnivores. The virus is a single-stranded RNA virus from the genus Morbillivirus and the family Paramyxoviridae. While domestic dogs are the most common hosts, the virus poses a significant threat to endangered wildlife due to its broad host range. This study aimed to characterize the CDV Haemagglutinin (H) gene in golden jackals and explore the molecular evolution of the virus in an underrepresented host. A total of 88 brain samples from hunted golden jackals were tested for the presence of CDV viral nucleic acid, and the H gene of positive samples was amplified and sequenced using the Sanger method. Phylogenetic analysis, conducted using maximum likelihood methods, revealed that all Serbian sequences clustered within the Arctic lineage. Notably, the analysis identified a tyrosine (Y) at position 549 of the H protein, a mutation commonly associated with wildlife hosts, instead of the histidine (H) typically found in domestic strains. Additionally, a mutation at position 310 was observed, which could potentially affect the protein’s function and virus-host interactions. These findings provide valuable insights into the genetic diversity and evolutionary dynamics of CDV in golden jackals, with broader implications for understanding the virus’s adaptability to different hosts. Further research is needed to investigate the functional impact of these mutations, particularly their role in vaccine efficacy and disease transmission across wildlife and domestic species.

Canine distemper virus (CDV) is a highly contagious and often fatal disease affecting wild and domesticated carnivores. The virus is a single-stranded RNA virus from the genus Morbillivirus and the family Paramyxoviridae. While domestic dogs are the most common hosts, the virus poses a significant threat to endangered wildlife due to its broad host range. This study aimed to characterize the CDV Haemagglutinin (H) gene in golden jackals and explore the molecular evolution of the virus in an underrepresented host. A total of 88 brain samples from hunted golden jackals were tested for the presence of CDV viral nucleic acid, and the H gene of positive samples was amplified and sequenced using the Sanger method. Phylogenetic analysis, conducted using maximum likelihood methods, revealed that all Serbian sequences clustered within the Arctic lineage. Notably, the analysis identified a tyrosine (Y) at position 549 of the H protein, a mutation commonly associated with wildlife hosts, instead of the histidine (H) typically found in domestic strains. Additionally, a mutation at position 310 was observed, which could potentially affect the protein's function and virus-host interactions. These findings provide valuable insights into the genetic diversity and evolutionary dynamics of CDV in golden jackals, with broader implications for understanding the virus's adaptability to different hosts. Further research is needed to investigate the functional impact of these mutations, particularly their role in vaccine efficacy and disease transmission across wildlife and domestic species. [ABSTRACT FROM AUTHOR]

Canine distemper (CD) is a virulent disease caused by the canine distemper virus (CDV) in canines and mustelidaes with high mortality. The incidence of CDV is worldwide distribution and it has caused huge economic losses to multiple industries around the world. There are many studies investigating the prevalence of CD infection, but no comprehensive analysis of CDV infection in minks, foxes and raccoon dogs worldwide has therefore been carried out. The aim of this meta is to provide a comprehensive assessment of the prevalence of CDV infection in minks, foxes and raccoon dogs dogs through a meta-analysis of articles published from around the world. Data from 8,582 small carnivores in 12 countries were used to calculate the combined prevalence of CD. A total of 22.6% (1,937/8,582) of minks, foxes and raccoon dogs tested positive for CD. The prevalence was higher in Asia (13.8, 95% CI: 22.2-45.6), especially in South Korea (65.8, 95% CI: 83.3-95.8). Our study found that the incidence of CD was also associated with geographic climate, population size, health status, and breeding patterns. CD is more commonly transmitted in minks, foxes and raccoon dogs. However, the concentrated breeding as an economic animal has led to an increase in the prevalence rate. The difference analysis study recommended that countries develop appropriate preventive and control measures based on the prevalence in the minks, foxes, and raccoon dogs industries, and that reducing stocking density is important to reduce the incidence of CDV. In addition, CDV is more common in winter, so vaccination in winter should be strengthened and expanded to reduce the incidence of CD in minks, foxes and raccoon dogs. [ABSTRACT FROM AUTHOR]

This review provides an overview of the canine distemper virus (CDV), a highly infectious pathogen causing severe disease in domestic dogs and wildlife. It shares genetic similarities with the human measles virus (HMV) in humans and the rinderpest virus (RPV) in cattle. The origin of CDV likely involves a mutation from human measles strains, possibly in the New World, with subsequent transmission to dogs. CDV has been globally observed, with an increasing incidence in various animal populations. Genomic mutations, especially in the H protein, contribute to its ability to infect different hosts. Diagnosis by molecular techniques like RT-qPCR offers rapid and sensitive detection when compared with serological tests. Genomic sequencing is vital for understanding CDV evolution and designing effective control strategies. Overall, CDV poses a significant threat, and genomic sequencing enhances our ability to manage and prevent its spread. Here, the epidemiology of CDV principally in Mexico is reviewed. [ABSTRACT FROM AUTHOR]

Oncolytic viruses and morbilliviruses in particular, represent an interesting therapeutic approach for tumors with a poor prognosis and frequent resistance to conventional therapies. Canine histiocytic sarcomas (HS) exemplify such a neoplasm in need for new curative approaches. Previous investigations demonstrated a limited success of an acute intratumoral application of canine distemper virus (CDV) on xenotransplanted canine histiocytic sarcoma cells (DH82 cells), while persistently CDV-infected DH82 cell transplants exhibited a complete spontaneous regression. Therefore, the present study focuses on an intratumoral application of persistently CDV vaccine strain Onderstepoort-infected DH82 (DH82 Ond p.i.) cells into non-infected subcutaneous DH82 cell transplants in a murine model. DH82 cell transplants that received 10 applications, two days apart, showed a transient growth retardation as well as larger areas of intratumoral necrosis, lower mitotic rates, and a decreased intratumoral vascularization compared to controls. Viral mRNA was detected in all neoplasms following application of DH82 Ond p.i. cells until 66 days after the last injection. Furthermore, infectious virus was present until 62 days after the last injection. Although complete regression was not achieved, the present application regimen provides promising results as a basis for further treatments, particularly with genetically modified viruses, to enhance the observed effects. [ABSTRACT FROM AUTHOR]

RNA viruses adapt rapidly to new host environments by generating highly diverse genome sets, so-called “quasispecies.” Minor genetic variants promote their rapid adaptation, allowing for the emergence of drug-resistance or immune-escape mutants. Understanding these adaptation processes is highly relevant to assessing the risk of cross-species transmission and the safety and efficacy of vaccines and antivirals. We hypothesized that genetic memory within a viral genome population facilitates rapid adaptation. To test this, we investigated the adaptation of the Morbillivirus canine distemper virus to ferrets and compared an attenuated, Vero cell-adapted virus isolate with its recombinant derivative over consecutive ferret passages. Although both viruses adapted to the new host, the reduced initial genetic diversity of the recombinant virus resulted in delayed disease onset. The non-recombinant virus gradually increased the frequencies of beneficial mutations already present at very low frequencies in the input virus. In contrast, the recombinant virus first evolved de novo mutations to compensate for the initial fitness impairments. Importantly, while both viruses evolved different sets of mutations, most mutations found in the adapted non-recombinant virus were identical to those found in a previous ferret adaptation experiment with the same isolate, indicating that mutations present at low frequency in the original virus stock serve as genetic memory. An arginine residue at position 519 in the carboxy terminus of the nucleoprotein shared by all adapted viruses was found to contribute to pathogenesis in ferrets. Our work illustrates the importance of genetic diversity for adaptation to new environments and identifies regions with functional relevance. IMPORTANCE When viruses encounter a new host, they can rapidly adapt to this host and cause disease. How these adaptation processes occur remains understudied. Morbilliviruses have high clinical and veterinary relevance and are attractive model systems to study these adaptation processes. The canine distemper virus is of particular interest, as it exhibits a broader host range than other morbilliviruses and frequently crosses species barriers. Here, we compared the adaptation of an attenuated virus and its recombinant derivative to that of ferrets. Pre-existing mutations present at low frequency allowed faster adaptation of the non-recombinant virus compared to the recombinant virus. We identified a common point mutation in the nucleoprotein that affected the pathogenesis of both viruses. Our study shows that genetic memory facilitates environmental adaptation and that erasing this genetic memory by genetic engineering results in delayed and different adaptation to new environments, providing an important safety aspect for the generation of live-attenuated vaccines. [ABSTRACT FROM AUTHOR]

Brucellosis is a global problem, with the causative agent being the genus Brucella. B. canis can cause undulant fever in dogs, which is a zoonotic disease that can spread not only among dogs but also to humans. This poses a public health threat to society. In this study, a rapid and straightforward immune colloidal gold test strip was developed for the diagnosis of canine brucellosis through the detection of anti-LPS antibodies in serum samples. Rabbit anti-canine IgG conjugated with colloidal gold was employed as the colloidal gold-labeled antibody. The extracted high-purity R-LPS was employed as the capture antigen in the test line (T-line), while goat anti-rabbit IgG was utilized as the capture antibody in the control line (C-line). The colloidal gold strip exhibited high specificity in the detection of brucellosis, with no cross-reaction observed with the common clinical canine diseases caused by Canine coronavirus (CCV), Canine distemper virus (CDV), and Canine parvovirus (CPV). In comparison to the commercial iELISA kit, the sensitivity and specificity of the colloidal gold test strip were found to be 95.23% and 98.76%, respectively. The diagnostic coincidence rate was 98.47%. The findings of this study indicate that colloidal gold test strips may be employed as a straightforward, expeditious, sensitive, and specific diagnostic instrument for the identification of canine brucellosis, particularly in resource-limited regions. [ABSTRACT FROM AUTHOR]

Canine distemper virus (CDV) belongs to morbillivirus, including measles virus (MeV) and rinderpest virus, which causes serious immunological and neurological disorders in carnivores, including dogs and rhesus monkeys, as recently reported, but their vaccines are highly effective. The attachment glycoprotein hemagglutinin (CDV-H) at the CDV surface utilizes signaling lymphocyte activation molecule (SLAM) and Nectin-4 (also called poliovirus-receptor-like-4; PVRL4) as entry receptors. Although fusion models have been proposed, the molecular mechanism of morbillivirus fusion entry is poorly understood. Here, we determined the crystal structure of the globular head domain of CDV-H vaccine strain at 3.2 Å resolution, revealing that CDV-H exhibits a highly tilted homodimeric form with a six-bladed ß-propeller fold. While the predicted Nectin-4-binding site is well conserved with that of MeV-H, that of SLAM is similar but partially different, which is expected to contribute to host specificity. Five N-linked sugars covered a broad area of the CDV-H surface to expose receptor-binding sites only, supporting the effective production of neutralizing antibodies. These features are common to MeV-H, although the glycosylation sites are completely different. Furthermore, real-time observation using high-speed atomic force microscopy revealed highly mobile features of the CDV-H dimeric head via the connector region. These results suggest that sugar-shielded tilted homodimeric structure and dynamic conformational changes are common characteristics of morbilliviruses and ensure effective fusion entry and vaccination. [ABSTRACT FROM AUTHOR]

Canine distemper virus (CDV) affects many domestic and wild animals. Variations among CDV genome linages could lead to vaccination failure. To date, there are several vaccine alternatives, such as a modified live virus and a recombinant vaccine; however, most of these alternatives are based on the ancestral strain Onderstepoort, which has not been circulating for years. Vaccine failures and the need to update vaccines have been widely discussed, and the development of new vaccine candidates is necessary to reduce circulation and mortality. Current vaccination alternatives cannot be used in wildlife animals due to the lack of safety data for most of the species, in addition to the insufficient immune response against circulating strains worldwide in domestic species. Computational tools, including peptide-based therapies, have become essential for developing new-generation vaccines for diverse models. In this work, a peptide-based vaccine candidate with a peptide library derived from CDV H and F protein consensus sequences was constructed employing computational tools. The molecular docking and dynamics of the selected peptides with canine MHC-I and MHC-II and with TLR-2 and TLR-4 were evaluated. In silico safety was assayed through determination of antigenicity, allergenicity, toxicity potential, and homologous canine peptides. Additionally, in vitro safety was also evaluated through cytotoxicity in cell lines and canine peripheral blood mononuclear cells (cPBMCs) and through a hemolysis potential assay using canine red blood cells. A multiepitope CDV polypeptide was constructed, synthetized, and evaluated in silico and in vitro by employing the most promising peptides for comparison with single CDV immunogenic peptides. Our findings suggest that predicting immunogenic CDV peptides derived from most antigenic CDV proteins could aid in the development of new vaccine candidates, such as multiple single CDV peptides and multiepitope CDV polypeptides, that are safe in vitro and optimized in silico. In vivo studies are being conducted to validate potential vaccines that may be effective in preventing CDV infection in domestic and wild animals. [ABSTRACT FROM AUTHOR]

Background: Canine distemper virus (CDV) is a pathogen with the capability of cross-species transmission. It has crossed the species barrier to infect many other species, and its host range is expanding. The reverse genetic platform, a useful tool for scientific research, allows the generation of recombinant viruses from genomic cDNA clones in vitro. Methods: To improve the reverse genetic system of CDV, a plasmid containing three independent expression cassettes was constructed for co-expression of the N, P, and L genes and then transfected with a full-length cDNA clone of CDV into Vero cells. Results: The results indicated that the established rescue system has the advantages of being more convenient, easy to control the transfection ratio, and high rescue efficiency compared with the conventional reverse genetics system. Conclusion: This method not only reduces the number of transfection plasmids, but also improves the rescue efficiency of CDV, which could provide a reference for the recovery of other morbilliviruses. [ABSTRACT FROM AUTHOR]

RNAscope in situ hybridization (ISH) detects target RNA in formalin-fixed, paraffin-embedded (FFPE) tissues. Protocols suggest that prolonged FFPE storage and formalin fixation may impact signal detection, potentially limiting the utility of RNAscope ISH in retrospective studies. To develop parameters for RNAscope use with archived specimens, we evaluated the effect of formalin-fixation time by measuring the signal of a reference gene (16srRNA) in selected tissues fixed in 10% neutral-buffered formalin for 1, 2, 3, 5, 7, 10, 14, 21, 28, 60, 90, 180, and 270 d. The signal intensity and percent area of signal decreased after 180 d. Tissues had detectable signal at 180 d but not at 270 d of formalin fixation. To assess target detection in paraffin blocks, we qualitatively compared the signal of canine distemper virus (CDV) antigen via immunohistochemistry and CDV RNA via RNAscope ISH in replicate sections from blocks stored at room temperature for 6 mo, 1, 3, 6, 8, 11, 13, and 15 y; RNA was detected in FFPE tissues stored for up to 15 y. Our results demonstrate that RNAscope ISH can detect targets in tissues with prolonged paraffin storage intervals and formalin-fixation times. [ABSTRACT FROM AUTHOR]

Canine distemper virus (CDV) is a highly contagious and often fatal virus that affects members of the Canidae family. Foxes are susceptible to CDV, and outbreaks among fox populations have been reported in several species. The symptoms of distemper in foxes are similar to those in domestic dogs. Foxes may contract the virus directly from infected animals or via exposure to a contaminated environment. Among domestic animals’ dogs are often a source of infection for domestic and wildlife populations, while among wildlife primary sources are other wildlife through direct contact or vectors such as insects. This review comprehensively discusses the virus itself, its pathology, clinical signs, diagnostic methods, treatment options, preventive measures, and its impact on fox populations. Wildlife veterinarians and researchers monitor and study the impact of diseases like distemper on fox populations. Understanding the spread and prevalence of such diseases is crucial for wildlife conservation efforts.

Abstract Background Canine distemper virus (CDV) is a pathogen with the capability of cross-species transmission. It has crossed the species barrier to infect many other species, and its host range is expanding. The reverse genetic platform, a useful tool for scientific research, allows the generation of recombinant viruses from genomic cDNA clones in vitro. Methods To improve the reverse genetic system of CDV, a plasmid containing three independent expression cassettes was constructed for co-expression of the N, P, and L genes and then transfected with a full-length cDNA clone of CDV into Vero cells. Results The results indicated that the established rescue system has the advantages of being more convenient, easy to control the transfection ratio, and high rescue efficiency compared with the conventional reverse genetics system. Conclusion This method not only reduces the number of transfection plasmids, but also improves the rescue efficiency of CDV, which could provide a reference for the recovery of other morbilliviruses.

Canine distemper virus (CDV) is a highly contagious and potentially lethal virus that affects dogs and other members of the Canidae family, including wolves, foxes, and coyotes. Here, we present a fluorescent lateral flow immunoassay (FLFA) platform for the detection of CDV, which utilizes fluorescent microspheres - fusion protein monoclonal antibody (mAb)-labeled monoclonal antibody. The assay detected CDV within 5  min, with a detection limit threshold of 3  ×  10² TCID50/mL. Notably, the assay demonstrated no cross-reactivity with canine parvovirus, canine coronavirus, canine adenovirus, feline calicivirus, feline herpesvirus, or feline parvovirus. Field and clinical applicability of the assay was evaluated using 63 field samples, including 30 canine fecal samples, 18 swab samples, and 15 blood samples. The coincidence rate between the detection results of clinical samples obtained through FLFA and reverse transcription polymerase chain reaction (RT-PCR) was 96.83%. Thus, this assay offers a significant advancement for the rapid diagnosis of CDV at the point of care. [ABSTRACT FROM AUTHOR]

Background: Canine coronavirus (CCoV), canine rotavirus (CRV), canine parvovirus (CPV), and canine distemper virus (CDV) cause gastroenteritis in dogs, and co-infections of these pathogens are common in China. In particular, CCoV and CRV are confirmed to have important zoonotic potential and cause public health issues. It is difficult to diagnose these diseases based only on clinical manifestations and pathological damage. Methods: In this study, four pairs of specific primers and probes targeting the CCoV M, CRV VP7, CPV VP2, and CDV N genes were designed. The reaction conditions, including the primer and probe concentrations, annealing temperatures, and reaction cycles, were optimized for the development of a quadruplex RT-qPCR for the detection of CCoV, CRV, CPV, and CDV. The assay was used to test 1028 clinical samples to validate its application. Results: A quadruplex RT-qPCR was successfully established for the differential detection of CCoV, CRV, CPV, and CDV, with good specificity, high sensitivity, and excellent repeatability. The assay could specifically detect CCoV, CRV, CPV, and CDV without cross-reactivity with the other canine viruses tested. It showed high sensitivity with limits of detection (LOD) of 1.1 × 102 copies/reaction for all four plasmid constructs. It showed excellent repeatability, with 0.05–0.90% intra-assay variation and 0.02–0.94% inter-assay variation. The 1028 clinical samples were tested using the quadruplex RT-qPCR and a reported reference RT-qPCR. The positivity rates of CCoV, CRV, CPV, and CDV were 9.53%, 0.97%, 25.68%, and 5.06% using the developed assay, and 9.05%, 0.88%, 25.68%, and 4.86% using the reference assay, with agreements higher than 99.32%. Conclusion: The results indicated that a rapid and accurate quadruplex RT-qPCR was developed for the detection and differentiation of CCoV, CRV, CPV, and CDV. [ABSTRACT FROM AUTHOR]

Simple Summary: Due to its promising applications in diagnosis and therapy, amniotic fluid may represent the substrate of the future in obstetric and regenerative medicine. In this study, we explored its potential impact on canine neonatal immunity by investigating, in both maternal plasma and amniotic fluid collected at birth, total and specific immunoglobulins G against the three viruses responsible for most of the neonatal mortalities in dogs: canine parvovirus (CPV-2), infectious canine hepatitis virus (CadV-1), and canine distemper virus (CDV). Our findings revealed that both total and specific plasma maternal IgG titres were not strictly related to vaccination status, whereas specific immunoglobulin G concentrations in amniotic fluids showed some correlation with the bitch vaccination status. Furthermore, puppies that developed pathological conditions (i.e., diarrhoea of any origin) within the first two months of life exhibited significantly lower amniotic CAdV-1 antibody titres compared to healthy ones. The evaluation of antibodies in amniotic fluid at birth could provide crucial information on the actual immune status of newborns. There is a growing interest in the composition of amniotic fluid (AF) in both humans and animals. In addition to its nutritional and protective functions for the foetus, current knowledge demonstrates that AF also serves advanced diagnostic, prognostic, and therapeutic roles. Newborn dogs have an underdeveloped immune system, making them highly susceptible to dangerous pathogens such as canine parvovirus (CPV-2), canine infectious hepatitis virus (CAdV-1), and canine distemper virus (CDV), thus exposing them to a high risk of mortality in the first weeks of life. Immunoglobulins G (IgGs) represent the only antibody isotype capable of crossing the placenta in a small amount and have been detected also in canine AF. The primary aim of this study was to investigate the reliability of AF collected at birth as a marker of passive immunity in canine species. For this purpose, total and specific IgGs against CPV-2, CAdV-1, and CDV were investigated and quantified in both maternal plasma and AF collected at the time of caesarean section. The vaccination status of the bitches was also taken into consideration. Since the immune system can be influenced by gestational age, with preterm infants having immature innate and adaptive immunity, IgG concentrations were correlated with amniotic lecithin, sphingomyelin, cortisol, surfactant protein A, and pentraxin 3 levels. In a previous study from our group on foetal maturity these molecules were measured in the same samples. Finally, correlations between their amniotic content and neonatal outcomes were investigated. This study demonstrates that AF analysis at birth can provide valuable insights into neonatal immunity in puppies, offering a non-invasive method to detect potential early health risks, for improved puppy care and management. [ABSTRACT FROM AUTHOR]

• The immune response to vaccination is moderately heritable in dogs. • Many SNPs of low to high effect contribute to vaccine-induced immune response. • Genes involving the spliceosome may play a role in vaccine-induced immune response. Both genetic and non-genetic factors contribute to individual variation in the immune response to vaccination. Understanding how genetic background influences variation in both magnitude and persistence of vaccine-induced immunity is vital for improving vaccine development and identifying possible causes of vaccine failure. Dogs provide a relevant biomedical model for investigating mammalian vaccine genetics; canine breed structure and long linkage disequilibrium simplify genetic studies in this species compared to humans. The objective of this study was to estimate the heritability of the antibody response to vaccination against viral and bacterial pathogens, and to identify genes driving variation of the immune response to vaccination in Beagles. Sixty puppies were immunized following a standard vaccination schedule with an attenuated combination vaccine containing antigens for canine adenovirus type 2, canine distemper virus, canine parainfluenza virus, canine parvovirus, and four strains of Leptospira bacteria. Serum antibody measurements for each viral and bacterial component were measured at multiple time points. Heritability estimations and GWAS were conducted using SNP genotypes at 279,902 markers together with serum antibody titer phenotypes. The heritability estimates were: (1) to Leptospira antigens, ranging from 0.178 to 0.628; and (2) to viral antigens, ranging from 0.199 to 0.588. There was not a significant difference between overall heritability of vaccine-induced immune response to Leptospira antigens compared to viral antigens. Genetic architecture indicates that SNPs of low to high effect contribute to immune response to vaccination. GWAS identified two genetic markers associated with vaccine-induced immune response phenotypes. Collectively, these findings indicate that genetic regulation of the immune response to vaccination is antigen-specific and influenced by multiple genes of small effect. [ABSTRACT FROM AUTHOR]

Canine coronavirus (CCoV) is a common agent of gastroenteritis in dogs, although some variants have been found associated with systemic and often fatal diseases. Distinct genotypes (CCoV-I and CCoV-II) and subgenotypes (CCoV-IIa and CCoV-IIb) are worldwide distributed. In Italy, CCoV infections have been occasionally evaluated, but information about the molecular epidemiology and the genomic features of currently circulating strains is limited. This study reports the detection and molecular characterization of CCoV strains from samples collected from 284 dogs in Italy between 2019 and 2021. CCoV RNA was detected in 39 (13.7%) dogs, as a single viral agent (5 animals, 12.8%) or with other viral pathogens (canine parvovirus types 2a/2b/2c; canine adenovirus type 1; norovirus GIV.2) (34 animals, 87.2%). A total of 48 CCoV strains were detected either alone (CCoV-I: 51.3%, CCoV-IIa: 20.5%) or in copresence (CCoV-I and CCoV-IIa, 23.1%); surprisingly, CCoV-IIb was not identified in this study. Five clusters of CCoV-I were detected, and their spike gene sequences showed the highest nucleotide identities with CCoV-I strains collected from Greece in 2008/2009 and from China in 2021. CCoV-IIa spike gene sequences (three variants) had the highest nucleotide identities with CCoV-IIa strains collected in Greece in 2008/2009 and in Italy in 2009/2011. Given the high CCoV diversity and the variable pathogenicity potential, we underline the need of further surveillance studies to increase our understanding of the epidemiology and evolution of these viruses. [ABSTRACT FROM AUTHOR]

The purpose of the experiment was to study the feasibility of fox carcass as meat and bone meal and evaluate the safety of fox meat and bone meal. Three healthy blue foxes (39 in total) weighing about 15 kg were randomly selected from 13 fox breeding areas in China. Fox carcass (including viscera and toe claws) was made into meat and bone meal by grinding, high-pressure hydrolysis, degreasing, drying, and crushing. Routine nutrients, heavy metals, veterinary drug residues, and canine distemper virus were determined, respectively, and the safety was evaluated. The results showed that the content of crude protein in meat and bone meal was between 55% and 67%, which meets the requirements of crude protein in the first-class index of Bone Meal and Meat and Bone Meal for Feed. The concentrations of lead, arsenic, mercury, cadmium, and chromium meet the requirements of the national standard for the hygiene and safety standards of meat and bone meal. Sulfamethazine, sulfamethoxazole, sulfamethoxazine, sulfadiazine, penicillin, oxytetracycline, and gentamicin meet the standard requirements of maximum residue limits of veterinary drugs in food. The results showed that fox meat and bone meal was rich in nutritional value, and its hygiene and safety indexes met the raw material standard of protein feed. [ABSTRACT FROM AUTHOR]

Background: Canine distemper (CD) is a worldwide spread disease that has been described in 12 families of mammals, especially in the Carnivora order, being better studied in domestic canines where vaccination represents the best means of control. CD is controlled by vaccination, but many cases of the disease still occur in vaccinated animals. Aim: The aim of this work was to study antigen-specific epitopes that can subsidize the development of a new vaccine approach. Methods: Mapping of T cell reactive epitopes for CD virus (CDV) was carried out through enzyme-linked immunospot assays using 119 overlapped synthetic peptides from the viral hemagglutinin protein, grouped in 22 pools forming a matrix to test the immune response of 32 animals. Results: Evaluations using the criteria established to identify reactive pools, demonstrated that 26 animals presented at least one reactive pool, that one pool was not reactive to any animal, and six pools were the most frequent among the reactive peptides. The crisscrossing of the most reactive pools in the matrix revealed nine peptides considered potential candidate epitopes for T cell stimulation against the CDV and those were used to design an in-silico protein, containing also predicted epitopes for B cell stimulation, and further analyzed using immune epitope databases to ensure protein quality and stability. Conclusion: The final in silico optimized protein presents characteristics that qualify it to be used to develop a new prototype epitope-based anti-CDV vaccine. [ABSTRACT FROM AUTHOR]

In this study, virological surveillance focused on coronaviruses in marmots in the Alpine region in 2022, captured as part of a population control reduction program in the Livigno area. Seventy-six faecal samples were randomly collected from marmots at the time of capture and release and tested for genome detection of pan-coronavirus, pan-pestivirus, canine distemper virus, and influenza A and D virus. Nine faecal samples were positive in the Pan-CoV RT-PCR, while all were negative for the other viruses. Pan-coronavirus positives were further identified using Illumina's complete genome sequencing, which showed the highest homology with Bovine Coronavirus previously detected in roe deer in the Alps. Blood samples (n.35) were collected randomly from animals at release and tested for bovine coronavirus (BCoV) antibodies using competitive ELISA and VNT. Serological analyses revealed that 8/35 sera were positive for BCoV antibodies in both serological tests. This study provides molecular and serological evidence of the presence of BCoV in an alpine marmot population due to a likely spillover event. Marmots share areas and pastures with roe deer and other wild ruminants, and environmental transmission is a concrete possibility. [ABSTRACT FROM AUTHOR]

Canine distemper virus (CDV) can cause fatal infections in giant pandas. Vaccination is crucial to prevent CDV infection in giant pandas. In this study, two bacterium-like particle vaccines F3-GEM and H4-GEM displaying the trimeric F protein or tetrameric H protein of CDV were constructed based on the Gram-positive enhanced-matrix protein anchor (GEM-PA) surface display system. Electron microscopy and Western blot results revealed that the F or H protein was successfully anchored on the surface of GEM particles. Furthermore, one more bacterium-like particle vaccine F3 and H4-GEM was also designed, a mixture consisting of F3-GEM and H4-GEM at a ratio of 1:1. To evaluate the effect of the three vaccines, mice were immunized with F3-GEM, H4-GEM or F3 and H4-GEM. It was found that the level of IgG-specific antibodies and neutralizing antibodies in the F3 and H4-GEM group was higher than the other two groups. Additionally, F3 and H4-GEM also increased the secretion of Th1-related and Th2-related cytokines. Moreover, F3 and H4-GEM induce IgG and neutralizing antibodies' response in dogs. Conclusions: In summary, F3 and H4-GEM can provoke better immune responses to CDV in mice and dogs. The bacterium-like particle vaccine F3 and H4-GEM might be a potential vaccine candidate for giant pandas against CDV infection. [ABSTRACT FROM AUTHOR]

Many different animal species are susceptible to SARS-CoV-2, including a few Canidae (domestic dog and raccoon dog). So far, only experimental evidence is available concerning SARS-CoV-2 infections in red foxes (Vulpes vulpes). This is the first report of SARS-CoV-2 RNA detection in a sample from a red fox. The RT-qPCR-positive fox was zoo-kept together with another fox and two bears in the Swiss Canton of Zurich. Combined material from a conjunctival and nasal swab collected for canine distemper virus diagnostics tested positive for SARS-CoV-2 RNA with Ct values of 36.9 (E gene assay) and 35.7 (RdRp gene assay). The sample was analysed for SARS-CoV-2 within a research project testing residual routine diagnostic samples from different animal species submitted between spring 2020 and December 2022 to improve knowledge on SARS-CoV-2 infections within different animal species and investigate their potential role in a One Health context. Within this project, 246 samples from 153 different animals from Swiss zoos and other wild animal species all tested SARS-CoV-2 RT-qPCR and/or serologically negative so far, except for the reported fox. The source of SARS-CoV-2 in the fox is unknown. The fox disappeared within the naturally structured enclosure, and the cadaver was not found. No further control measures were undertaken. [ABSTRACT FROM AUTHOR]

Background: Canine distemper (CD) is the dogs' most contagious and lethal viral disease. Despite the widespread use of vaccines to control CD, the prevalence of the CD virus (CDV) has increased at an alarming rate in recent years. Objectives: To identify the genotypes responsible for the neurological and non-neurological clinical forms of CD and to investigate the presence of the virus in the neurological and non-neurological forms of the disease. Methods: In this descriptive-analytical study, the samples were collected from 70 CD suspected unvaccinated dogs with clinical signs of distemper. All cases were tested with rapid tests and separated into 3 groups based on clinical signs and symptoms. Cerebral spinal fluid (CSF), respiratory secretion, and fecal samples of allall 70 cases were examined for reverse transcription polymerase chain reaction (RT-PCR). After sequencing the hemagglutinin gene (H gene), phylogenetic analysis of the gene isolated from CDVs was carried out using MEGA software, version 7. Results: The RT-PCR results showed that the respiratory secretion sample in the non-neurological CDV group (85%) and the neurological CDV group (80%) had the highest level of virus contamination. However, in the non-neurological CDV group, the CSF sample (40%) had a high level of infection. In neurotic groups, cases older than 12 months showed the highest percentage of distemper contamination, and in the non-neurologic CDV group, those between 3 and 6 months were more involved. Sequencing and phylogenetic analysis of the H gene revealed the CDV as a member of the endemic Arctic-like genetic lineage. Conclusion: The genotypic examination of the hemagglutinin gene of the distemper virus reveals that the recent isolates of neurologic and non-neurologic clinical forms of CDV in Iran are similar. In positive rapid test cases, the PCR test of respiratory secretions for virus detection ranks with the highest sensitivity. In neurologic cases with negative rapid test results, PCR of CSF had the highest sensitivity, so it may serve as a diagnostic criterion. [ABSTRACT FROM AUTHOR]

Taenia crassiceps is a zoonotic tapeworm of the genus Taenia that is distributed throughout the Northern Hemisphere. Wild and domestic carnivores are final hosts, while rodents and rabbits are primarily intermediate hosts, although many other mammals may harbour the larval stage, Cysticercus longicollis. This case report aims to describe C. longicollis infection in a lemur and molecularly characterise the isolated parasite. The excised lesion was subjected to morphological and histopathological examination, which revealed cysticerci of the tapeworm. Formalin-fixed and paraffin-embedded block (FFPEB), as well as the cysticerci fixed with formalin stored for one year, were subjected to molecular analysis, which aimed at detecting the partial mitochondrial cytochrome c oxidase subunit 1 (cox1) gene of Taenia sp. Based on the morphological characteristics, the parasite was identified as a metacestode of T. crassiceps. The presence of the cox1 gene was detected using polymerase chain reaction (PCR) in all samples. A randomly selected PCR product was sequenced and compared with other sequences from the GenBank database, confirming that the detected parasite was T. crassiceps. This article reports the first case of T. crassiceps cysticercosis in a lemur (Lemur catta) in Croatia and emphasises the potential risk of transmission from wild carnivores. [ABSTRACT FROM AUTHOR]

Objective: Canine distemper virus (CDV) is a highly infectious virus that represents a threat for domestic dogs and several wild species. Despite recognized in several African countries, current knowledge of its molecular epidemiology is scarce and poorly updated. Design: Twenty-two hemagglutinin sequences, obtained from symptomatic Namibian dogs from 2020 to 2023, were analysed through phylogenetic and phylodynamic analysis to characterize the local CDV epidemiology and contextualize it in the international scenario. Results: Two unrelated clades were identified, including strains sampled in different Namibian towns, in the absence of a strong geographical clustering. The ancestors of the two clades were estimated to have originated from South America, likely Brazil, and South Africa, approximately in 2000 and 2006, respectively. While the introduction from South Africa was predictable, the introduction from Brazil was unexpected. The mediation of other African countries, particularly Angola, appears to be the most likely importation pathway. Conclusions: The occurrence of multiple introduction events, likely originating from cross-border illegal animal trade between African countries, and the absence of any geographical clustering within Namibian regions, suggest a need for further investigation into its spreading pattern, as well as improved biosecurity measures to limit foreign viral introduction into the country.

Since the end of 2019, COVID-19 pandemic caused by the novel SARS-CoV-2 has become a serious problem for the world. Accurate and rapid techniques in testing and tracing are needed to control the virus spreading. Molecular diagnostics through gene amplification techniques, especially PCR, still become the gold standard for SARS-CoV-2 detection, which requires the first step of RNA extraction and purification. The limitations of commercial RNA extraction-purification kits during the pandemic caused a big problem in testing and tracing, especially for developing countries. A simple RNA extraction-purification kit based on magnetic-silica (MAGSi) beads and non-guanidine in-house buffers for RNA virus extraction-purification has been developed. Two types of MAGSi beads with different magnetic nanoparticles (MNPs) content were synthesized through a modified Stöber’s method using the sonication technique. The PCR result shows that both the MAGSi beads and the buffer can be used as a kit for RNA extraction-purification, tested for SARS-CoV-2 and Canine Distemper Virus. Further study shows that MAGSi-1 has better RNA extraction ability, and a higher concentration of RNA has been extracted. This is likely because of the smaller particle size distribution (50–1,500 nm distribution) and higher magnetization (20.2 emu/g) of MAGSi-1 compared to MAGSi-2 with 100–1,700 nm size distribution and 14.2 emu/g magnetization.

The article discusses research which performed the first complete meta-transcriptomic (MTT) genomic sequence analysis of canine distemper virus (CDV) in wild boar. The study analyzed tissue samples from free-ranging wild boars and determined the CDV infection rate using rapid amplification of complementary DNA (cDNA) ends and specific reverse transcription polymerase chain reaction (RT-PCR). It investigated CDV seroprevalence in wild boars in northeast China using virus-neutralization tests.

We describe a natural coinfection with canine distemper virus (CDV) and yellow fever virus in a free-ranging neotropical primate of the genus Callithrix, found dead in the northeastern region of Brazil. The laboratory diagnosis included histopathology, immunohistochemistry, rRT-PCR, and phylogenetic analyses. The CDV sequences from this primate in Brazil represent a divergent lineage in Rio Grande do Norte, closely related to genotypes EU1/South America 1 and South America 2. To our knowledge, this is the first report of natural coinfection by CDV and yellow fever virus in a neotropical primate, underscoring the need to further investigate the circulation of this virus in Brazilian nonhuman primates and its potential implications for wildlife conservation.

Canine distemper virus (CDV) poses a substantial threat to diverse carnivorans, leading to systemic and often fatal diseases. Accurate and prompt diagnosis is paramount for effective management and curbing further transmission. This study evaluates the diagnostic performance of droplet digital PCR (ddPCR) in comparison to conventional reverse-transcription (RT-PCR) and quantitative reverse-transcription real-time PCR (RT-qPCR). Seventy-six clinical samples were collected from dogs with CDV symptoms diagnosed by specialized veterinarians, and sixteen samples from apparently healthy individuals. Conventional PCR, quantitative real-time PCR, and ddPCR were deployed, and their diagnostic capabilities were meticulously assessed. DdPCR exhibited heightened analytical sensitivity, reaching a detection limit of 3 copies/μL, whereas RT-qPCR had a detection limit of 86 copies/μL. The comparative analysis between clinical diagnosis and molecular techniques, including RT-PCR and RT-qPCR, demonstrated low concordance, with Kappa coefficients of 0.268 and 0.324, respectively. In contrast, ddPCR showed a moderate concordance, with a Kappa coefficient of 0.477. The sensitivity was 42.4% for RT-PCR, 57.9% for RT-qPCR, and 72.4% for ddPCR, with 100% specificity for all methods. This study underscores ddPCR’s superior sensitivity and agreement with clinical CDV diagnosis, even at low viral concentrations, suggesting it as a promising alternative for CDV diagnosis.

Canine distemper (CD) is a deadly, multi-system infection caused by a Morbillivirus. The canine distemper virus (CDV) frequently affects the nervous system with demyelinating leukoencephalitis, the most common neurological lesion. The disease has been linked to multiple sclerosis (MS) in humans due to similar clinical presentation and pathophysiological mechanisms. In MS, visual evoked potentials (VEPs) have been identified as a reliable marker for disease progression, enabling the early detection of clinically suspected lesions. The aim of this study was to determine if there are any abnormalities in VEP responses in dogs with neurological CD. Visual evoked potentials and electroretinogram (ERG) were recorded at both the cranial and spinal levels in dogs naturally infected with CDV and in healthy dogs. The results in the CDV-infected group revealed a bilateral increase in the latency of N1, P1, N2, P2, and N3 waves of the VEPs, without any alterations in their amplitudes. No significant differences were observed in the ERG between the groups. These results suggest that altered VEP responses could serve as an early diagnostic indicator of neurological damage caused by distemper. Therefore, conducting these studies could potentially aid in the detection of central nervous conduction disorders during the subclinical phases of the disease.

Background: Morbillivirus canis is the etiological agent of a highly contagious disease that affects diverse domestic and wild animals. Vaccination is considered the most suitable strategy for controlling CDV dissemination, transmission, and distemper disease. However, the emergence of new CDV strains has led to the need to update the current vaccine strategies employed to prevent CDV infection in domestic and wild animals. Currently, there is a lack of effective alternatives for wild animals. Diverse computational tools, especially peptide-based therapies, enable the development of new universal vaccines. Objective: The aim of this study was to evaluate the safety and humoral and cellular immune response of a new generation of vaccines based on CDV peptides as single-peptide mixtures or multiepitope CDV polypeptides in mice. Methods: Twenty-four BALB/c mice were subjected to a three-dose regimen for 28 days. Seroconversion was evaluated via ELISA, and cellular immune responses were evaluated via flow cytometry through activation-induced markers (AIMs). Results: Compared with the placebo, the peptide mixture and multiepitope CDV polypeptide were safe, and seroconversion was statistically significant in the multiepitope CDV polypeptide and commercial vaccine (CV) groups. The numbers of antigen-specific CD4+CD134+ and IFN-γ+ T cells, CD8+ T cells and TNF-α- and IL-6-producing cells were greater in the mice immunized with the multiepitope CDV polypeptide than in the control mice. Conclusion: This combined approach represents a potential step forward in developing new immunization candidates or enhancing current commercial vaccines to control CDV disease in domestic dogs and wild animals.

Concurrent Clostridium piliforme and canine distemper virus (CDV) infection was diagnosed in 2 canine littermates and 1 gray fox kit from Texas, USA. In all 3 animals, intracytoplasmic, filamentous bacteria, consistent with C. piliforme, were present along the margins of foci of hepatic necrosis. Additional histologic findings included intracytoplasmic and intranuclear inclusion bodies in bile duct and bronchial epithelial cells of the fox kit, and mild intestinal necrosis in 1 puppy. PCR assays confirmed the presence of C. piliforme in all 3 animals, CDV in both puppies, and canine parvovirus in 1 puppy. Fluorescent antibody testing confirmed the presence of CDV in the fox kit. Concurrent canine distemper and Tyzzer disease in canine littermates and the gray fox has not been reported previously, to our knowledge.

Canine distemper virus (CDV), canine adenovirus type 2 (CAV-2), canine parvovirus (CPV), and canine parainfluenza virus 5 (CPIV-5) are the major viral pathogens in dogs. Despite the availability of vaccines for dogs against these 4 viral pathogens, investigations of antibodies against these pathogens have rarely been reported in South Korea. In this study, we investigated the recent incidence of viral diseases in dogs and conducted sero-surveillance for CDV, CAV-2, CPV, and CPIV-5 in Korean dogs. The most frequently diagnosed canine viral disease in Korean dog samples from 2000 to 2022 was CPV infection, which accounted for 48.7% (464/953) of the cases. A total of 400 dog serum samples collected between 2019 and 2022 were screened for the presence of virus-neutralizing antibodies against CDV, CAV-2, CPV, and CPIV-5. The overall seropositivity rates for CDV, CAV-2, CPV, and CPIV5 were 83.8%, 77.8%, 99.3%, and 82.0%, respectively. The protection rate against CPV was the highest (98.3%) and that against CAV-2 was the lowest (44.8%) in dog sera. Male and female dogs showed no significant differences in seropositivity rates. CDV and CPIV-5 seropositivity increased with age in dogs, and the highest incidence and seropositivity rates of CPV indicated that Korean dogs have been continuously exposed to wild CPV, and that CPV is a pathogen that urgently requires attention among canine viral diseases. [ABSTRACT FROM AUTHOR]

Taenia hydatigena, an intestinal parasite of domestic and wild canids which can cause medical problems and economic losses in animals. Thirty T. hydatigena recovered from the intestine of stray dogs after T. hydatigena based on morphological characters and morphometric observations involving Scanning Electron Microscopy (SEM) and the scoleces were stained by Semichon's acid carmine and viewed on a light microscope. High-quality SEM photographs were shown the blade dimensions of both large and small hooks (83.89 µm and 48.16 µm) respectively, which was close to measurements by a digital camera. The number of hooks around the rostellum were 30 hooks, and both were equal in number (15 hooks), while the average length of the large hooks was 220.11 ±28.07 µm, and was 88.58 ±7.92, and the average length of small hooks was142.37 ±12.52 µm and was 65.73 ±6.94 µm. body surface of Taenia hydatigena presence of characteristic transverse cuticular striations. [ABSTRACT FROM AUTHOR]

Morbilliviruses are members of the family Paramyxoviridae and are known for their ability to cause systemic disease in a variety of mammalian hosts. The prototypic morbillivirus, measles virus (MeV), infects humans and still causes morbidity and mortality in unvaccinated children and young adults. Experimental infection studies in non-human primates have contributed to the understanding of measles pathogenesis. However, ethical restrictions call for the development of new animal models. Canine distemper virus (CDV) infects a wide range of animals, including ferrets, and its pathogenesis shares many features with measles. However, wild-type CDV infection is almost always lethal, while MeV infection is usually self-limiting. Here, we made five recombinant CDVs, predicted to be attenuated, and compared their pathogenesis to the non-attenuated recombinant CDV in a ferret model. Three viruses were insufficient ly attenuated based on clinical signs, fatality, and systemic infection, while one virus was too attenuated. The last candidate virus caused a self-limiting infection associated with transient viremia and viral dissemination to all lymphoid tissues, was shed transiently from the upper respiratory tract, and did not result in acute neurological signs. Additionally, an in-depth phenotyping of the infected white blood cells showed lower infection percentages in all lymphocyte subsets when compared to the non-attenuated CDV. In conclusion, infection models using this candidate virus mimic measles and can be used to study pathogenesis-related questions and to test interventions for morbilliviruses in a natural host species. [ABSTRACT FROM AUTHOR]

The body of a 14-wk-old puppy (Canis familiaris) was submitted to the Animal Health Laboratory, University of Guelph, Ontario for postmortem examination following a history of intermittent anorexia and lethargy progressing to pyrexia, pruritic skin rash, mucoid nasal discharge, decreased mentation, dysphagia, muscle twitches, and focal seizures. Gross examination revealed rhinitis and pulmonary edema. Histologically, there was fibrinonecrotizing bronchopneumonia, tracheitis, and neutrophilic and lymphohistiocytic rhinitis; rarely within the cortical gray and white matter of the brain were small clusters of glial cells, with rare individual neutrophils in the choroid plexus. Although canine distemper was suspected, none of the usual supportive histologic lesions of distinct syncytial cells, viral inclusion bodies, or demyelinating leukoencephalitis were observed. Lung and brain tissues were PCR-positive for canine distemper virus (CDV), and CDV was detected immunohistochemically in the brain. The agent from the PCR-positive sample from the brain was genotyped and was a 99.9% match to the CDV Rockborn strain, indicating that the disease agent in our case was vaccinal in origin. Our unusual case highlights the possibility of reversion to virulence in a modified-live virus vaccine, and the occurrence of a disease in the absence of a full complement of the usual and compatible histologic lesions. [ABSTRACT FROM AUTHOR]

In recent years, plague has re-emerged in several countries around the world and remains endemic in some regions. In a natural environment and in contact with rodents and their fleas, stray carnivores are most at risk of catching the disease and maintaining the spread of the bacillus. The present study aimed to demonstrate the presence or absence of Yersinia pestis in stray dogs and cats in the Algiers region by molecular methods and thus determine their role in epidemiology of this disease. Molecular research of Yersinia pestis has also been conducted on fleas from these carnivores. Preliminary identification of ectoparasites to genus and species level was performed. Real-time polymerase chain reaction targeting Yersinia pestis pla gene was used to survey the plague agent in fleas and carnivores captured as stray animals in Algiers (Algeria). Positive qPCR results were tested by PCR sequencing using glpD gene. Among 327 fleas captured from 107 dogs and 365 fleas from 140 cats, prevalence of Ctenocephalides felis was higher in cats (86,96%), whereas that of Ctenocephalides canis and Xenopsylla cheopis were higher in dogs (90,57% and 92,63%, respectively). While internal and external PCR positive controls were positive, none of the 107 dogs spleens and 140 cats spleens and none of the 256 analyzed fleas were positive for Y. pestis. These results suggested that stray cats and dogs are unlikely sources of plague in Algeria, contrary to what has been reported in other plague-endemic countries. This observation illustrates that the plague epidemiological chain varies from one region to another. [ABSTRACT FROM AUTHOR]

Understanding mortality causes is important for the conservation of endangered species, especially in small and isolated populations inhabiting anthropized landscapes where both natural and human-caused mortality may hinder the conservation of these species. We investigated the mortality causes of 53 free-ranging brown bears (Ursus arctos) found dead between 1998 and 2023 in the Cantabrian Mountains (northwestern Spain), a highly human-modified region where bears are currently recovering after being critically threatened in the last century. We detected natural traumatic injuries in 52.63% and infectious diseases in 39.47% of the 38 bears for which the mortality causes were registered, with 21.05% of these cases presenting signs of both infectious diseases and traumas. More specifically, almost 30% of the bears died during or after intraspecific fights, including sexually selected infanticide (10.53%). In addition, primary infectious diseases such as infectious canine hepatitis, distemper, clostridiosis and colibacillosis caused the death of 15.79% of the bears. The number of direct human-caused deaths (i.e., shooting, poisoning, snare) decreased over the study period. This study also reveals three new mortality causes triggered by pathogens, two of which—Clostridium novyi and verotoxigenic Escherichia coli—not previously described in ursids, and the other one, canine distemper virus, never reported in brown bears as cause of death. New management strategies for the conservation of Cantabrian bears, which are urgently needed due to the rapid expansion of the population, should consider the mortality causes described in this study and must promote further research to elucidate how the high prevalence of infectious diseases may threaten the current recovery of the population. [ABSTRACT FROM AUTHOR]