SARS-CoV-2 wildlife surveillance in Ontario and Québec, Canada

BackgroundSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus responsible for the COVID-19 pandemic, is capable of infecting a variety of wildlife species. Wildlife living in close contact with humans are at an increased risk of SARS-CoV-2 exposure and if infected have the potential to become a reservoir for the pathogen, making control and management more difficult.ObjectiveTo conduct SARS-CoV-2 surveillance in urban wildlife from Ontario and Québec, Canada, increasing our knowledge of the epidemiology of the virus and our chances of detecting spillover from humans into wildlife.MethodsUsing a One Health approach, we leveraged activities of existing research, surveillance, and rehabilitation programs among multiple agencies to collect samples from 776 animals from 17 different wildlife species between June 2020 and May 2021. Samples from all animals were tested for the presence of SARS-CoV-2 viral RNA, and a subset of samples from 219 animals across 3 species (raccoons, Procyon lotor; striped skunks, Mephitis mephitis; and mink, Neovison vison) were also tested for the presence of neutralizing antibodies.ResultsNo evidence of SARS-CoV-2 viral RNA or neutralizing antibodies was detected in any of the tested samples.ConclusionAlthough we were unable to identify positive SARS-CoV-2 cases in wildlife, continued research and surveillance activities are critical to better understand the rapidly changing landscape of susceptible animal species. Collaboration between academic, public and animal health sectors should include experts from relevant fields to build coordinated surveillance and response capacity.

Semi-quantitative risk assessment of peste des petits ruminants introduction with wild animals into Russian Federation

The Russian Federation was officially recognized free from peste des petitts ruminants (PPR). As far as the disease infects both domestic and wild small ruminants, it is important to identify the level of the threat associated with the wild fauna diversity in the neighboring countries, where PPR outbreaks were reported. For that reason, habitats of various disease susceptible animal species were examined. Habitats of the wild susceptible animals were mapped for further examination of the interactions between different animal species using zoological research data; PPR outbreaks in wild animals were also designated in the map thus allowing for the detection of the potential routes of the infection spread in the population and introduction to the country. Analysis of the PPR epidemic situation in the country demonstrated that the disease cases were reported in wild mountain animals (ibices and moufflons) and migratory steppe animals (gazelles and saigas). Risk of this highly contagious viral disease spread in wild small ruminants in Mongolia was reported (probability 0.77). Expert survey was carried out for the determination of possible trends and factors of the infection introduction with the wild susceptible animals, through which small ruminant epizootologists assessed the risk probability. During the survey it was determined that PPR was expected to be introduced from Mongolia (probability 0.81), and of major significance were seasonal migrations of wild animal populations. The resulted semi-quantitative parameters of the potential risk can be recommended for the arrangement and implementation of measures aimed at prevention of PPR introduction and spread in the intact domestic and wild small ruminant populations inhabiting the territory of the Russian Federation.

Identification of the O/ME-SA/Ind-2001e Sublineage of Foot-and-Mouth Disease Virus in Cambodia

Foot-and-mouth (FMD) is endemic in Cambodia with numerous outbreaks in cattle, pigs and other susceptible animal species reported every year. Historically, these outbreaks were caused by the FMD virus (FMDV) of serotype O PanAsia and Mya-98 lineages and serotype A Sea-97 lineage. However, the trans-pool movement of FMDV between inter-pool regions or countries throughout FMD endemic regions has raised concerns regarding infection with the new genotype or serotype of FMDV in Cambodia. In this study, 19 sequences of VP1 coding region obtained from 33 clinical samples collected from FMDV-affected cattle farms in Cambodia during January to March 2019 were genetically characterized to identify the genotypes/lineages of FMDV.Phylogenetic analysis of VP1 coding sequences revealed that recent field viruses belonged to O/ME-SA/Ind-2001e (15.8%), O/ME-SA/PanAsia (52.7%), and A/ASIA/Sea-97 (31.5%). Besides, the field viruses of O/ME-SA/Ind-2001e in Cambodia showed 93.5–96.8% identity with the VP1 coding sequences of the same sublineage viruses from pool 1 and 2 surrounding Cambodia. This is the first report of O/ME-SA/Ind-2001e infection in Cambodia, suggesting that the trans-pool movement of the new genotype should be closely monitored for efficient control of FMD.

Evolutionary Dynamics and Epidemiology of Endemic and Emerging Coronaviruses in Humans, Domestic Animals, and Wildlife

Diverse coronavirus (CoV) strains can infect both humans and animals and produce various diseases. CoVs have caused three epidemics and pandemics in the last two decades, and caused a severe impact on public health and the global economy. Therefore, it is of utmost importance to understand the emergence and evolution of endemic and emerging CoV diversity in humans and animals. For diverse bird species, the Infectious Bronchitis Virus is a significant one, whereas feline enteric and canine coronavirus, recombined to produce feline infectious peritonitis virus, infects wild cats. Bovine and canine CoVs have ancestral relationships, while porcine CoVs, especially SADS-CoV, can cross species barriers. Bats are considered as the natural host of diverse strains of alpha and beta coronaviruses. Though MERS-CoV is significant for both camels and humans, humans are nonetheless affected more severely. MERS-CoV cases have been reported mainly in the Arabic peninsula since 2012. To date, seven CoV strains have infected humans, all descended from animals. The severe acute respiratory syndrome coronaviruses (SARS-CoV and SARS-CoV-2) are presumed to be originated in Rhinolopoid bats that severely infect humans with spillover to multiple domestic and wild animals. Emerging alpha and delta variants of SARS-CoV-2 were detected in pets and wild animals. Still, the intermediate hosts and all susceptible animal species remain unknown. SARS-CoV-2 might not be the last CoV to cross the species barrier. Hence, we recommend developing a universal CoV vaccine for humans so that any future outbreak can be prevented effectively. Furthermore, a One Health approach coronavirus surveillance should be implemented at human-animal interfaces to detect novel coronaviruses before emerging to humans and to prevent future epidemics and pandemics.

Study of enterobacteria biofilms critical point control technologies livestock and food production

The relevance of the study and the presence of gaps in the existing knowledge on the topic. Monitoring studies of the biological safety of food raw materials for microbiological indicators is an urgent problem due to the increase in the number registered diseases transmitted to humans through raw materials and products of animal origin. There is a tendency for a statistically significant increase in epidemiological indicators throughout the world, the proportion of these pathologies is increasing both in humane medicine and veterinary medicine. The aim of the work is a comparative assessment and selection of effective methods for studying the formation biofilms enterobacteriaceae circulating among susceptible animal species and isolated from food raw materials.Methods. Аnalysis of growth and dynamics development biofilms Enterobacteriaceae was carried out during cultivation on nutrient media containing growth factors for the repair of the cell wall and the reversal viable uncultured microorganisms. To study the morphological and functional patterns of the development a population microorganisms in vitro and in vivo, we used the conventional and developed methods for preparing preparations for scanning, transmission phase contrast, optical and luminescence microscopy.Results and its discussion. During microbiological control critical points in the technology of animal husbandry and food production, the morphological and functional characteristics biofilms, which are communities microorganisms secreting a polymer matrix and adhered to the tissues of susceptible animal species and abiotic surfaces livestock buildings and food industries, were studied. The developed methods of biofilm cultivation made it possible to study enterobacteriaceae biofilms in vitro and in vivo, without disturbing the natural architectonics of the population microorganisms, to determine the components extracellular matrix. For the study dynamics morphological and functional patterns of the development populations microorganisms, routine and technological advances present are recognized as promising, for example, scanning electron microscopy makes it possible to assess the degree of formation and morphological composition biofilms. Phase contrast microscopy to reveal processes depending on the composition medium and the oxygen content in the culture medium.Conclusions. Methods for cultivating biofilms in vitro and in vivo without disturbing the natural architectonics of biofilms made it possible to optimize the preparation samples for research and eliminate the routine stages of colony counting, and significantly increase the number of analyzes. Due to the simplicity of operations and minimization manual labor, productivity increases, safety of work is increased, cost of personnel working time is reduced, and subjective factors are excluded. For the development of a complex antiepizootic and diagnostic measures, a priority direction is the disclosure of scientific knowledge in the field fundamental studies of ecological plasticity and adaptation potentially pathogenic enterobacteria to parasitism in the warm-blooded organism of birds and mammals. This will allow solving applied problems controlling the critical points of livestock and food production technology, developing effective chemotherapeutic and disinfecting drugs to reduce cell coaggregation and detect viable uncultured microorganisms.

Analysis of Intermediate Hosts and Susceptible Animals of SARS-CoV-2 by Computational Methods

COVID-19, a disease caused by SARS-CoV-2 that produces major symptoms of pneumonia, has been a disaster worldwide. The traceability of SARS-CoV-2 and the discovery of susceptible animal species is crucial to halt viral transmission and explore the mechanism of cross-species transmission. We selected 82 representative ACE2 sequences from the 1000 sequences with the closest homology to the hACE2 protein. All selected ACE2 proteins were subjected to homology modeling. Potential natural and intermediate hosts, as well as animal species susceptible to SARS-CoV-2, were analyzed systematically by calculation of the binding free energy of ACE2 protein to the RBD of SARS-CoV-2. Primates, some wild Felidae, civets, goats, spotted hyenas and golden hamsters are susceptible to SARS-CoV-2 and may be potential intermediate hosts, whereas pangolins, birds and reptiles are unlikely to be intermediate hosts. Mice, rats and guinea pig are not susceptible to SARS-CoV-2. Given their possible susceptibility, non-human primates, goats and golden hamsters could potentially be used as experimental models to examine SARS-CoV-2 infection without transgenesis. Herein, possible candidates for the natural and intermediate hosts of SARS-CoV-2 are suggested, to provide guidance for subsequent studies.

In vitro Characterization of Fitness and Convalescent Antibody Neutralization of SARS-CoV-2 Cluster 5 Variant Emerging in Mink at Danish Farms

In addition to humans, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can transmit to animals that include hamsters, cats, dogs, mink, ferrets, tigers, lions, cynomolgus macaques, rhesus macaques, and treeshrew. Among these, mink are particularly susceptible. Indeed, 10 countries in Europe and North America reported SARS-CoV-2 infection among mink on fur farms. In Denmark, SARS-CoV-2 spread rapidly among mink farms and spilled-over back into humans, acquiring mutations/deletions with unknown consequences for virulence and antigenicity. Here we describe a mink-associated SARS-CoV-2 variant (Cluster 5) characterized by 11 amino acid substitutions and four amino acid deletions relative to Wuhan-Hu-1. Temporal virus titration, together with genomic and subgenomic viral RNA quantitation, demonstrated a modest in vitro fitness attenuation of the Cluster 5 virus in the Vero-E6 cell line. Potential alterations in antigenicity conferred by amino acid changes in the spike protein that include three substitutions (Y453F, I692V, and M1229I) and a loss of two amino acid residues 69 and 70 (ΔH69/V70), were evaluated in a virus microneutralization assay. Compared to a reference strain, the Cluster 5 variant showed reduced neutralization in a proportion of convalescent human COVID-19 samples. The findings underscore the need for active surveillance SARS-CoV-2 infection and virus evolution in susceptible animal hosts.

Susceptibility of animal species to experimental SARS-CoV-2 (Coronaviridae: Coronavirinae: Betacoronavirus; Sarbecovirus) infection

Due to the new coronavirus infection pandemic, the global scientific community has been forced to change the direction of the most research, focusing on vaccine development as well as the search for new antiviral drugs to treat COVID-19. The choice of experimental models, timeframe and approaches for evaluating drugs and vaccines under development is crucial for the development of effective measures to prevent and control this disease.The purpose of this review was to summarize the relevant data concerning the susceptibility of laboratory animals to SARS-CoV-2. This paper describes the most virus-susceptible animal species that can be used to reproduce coronavirus infection, stressing the main advantages and disadvantages of each of them.According to the latest data, small rodents (Rodentia) and non-human primates (Strepsirrhini) are commonly used in the scientific community to model coronavirus infection. The viral load in the upper and lower parts of the respiratory system, clinical symptoms of infection (weight loss, body temperature and general health status), pathomorphological picture in target organs and the production of antibodies after infection are considered to the main markers of pathology. Despite the vast amount of data, none of the described models of SARS-CoV-2 infection may be considered a gold standard, since they do not reproduce all spectrum of morphological and pathogenetic mechanisms of infection, and do not fully reflect the clinical picture observed in patients in human population.Based on the analyzed literature data, we suppose that Syrian hamster (Mesocricetus auratus) and mice (Muridae) expressing the angiotensin converting enzyme receptor 2 (ACE2) are the most suitable animal species for their use in experiments with SARS-CoV-2 infection. The development of neutralizing antibodies makes it possible to evaluate the efficacy of vaccines, while the course and severity of symptoms infection makes the use of mice and hamsters especially popular for screening pharmacological substances with antiviral mechanism of action, when their administration can prevent or slow the disease progression.

Current Status of Putative Animal Sources of SARS-CoV-2 Infection in Humans: Wildlife, Domestic Animals and Pets

SARS-CoV-2 is currently considered to have emerged from a bat coronavirus reservoir. However, the real natural cycle of this virus remains to be elucidated. Moreover, the COVID-19 pandemic has led to novel opportunities for SARS-CoV-2 transmission between humans and susceptible animal species. In silico and in vitro evaluation of the interactions between the SARS-CoV-2 spike protein and eucaryotic angiotensin-converting enzyme 2 (ACE2) receptor have tentatively predicted susceptibility to SARS-CoV-2 infection of several animal species. Although useful, these data do not always correlate with in vivo data obtained in experimental models or during natural infections. Other host biological properties may intervene such as the body temperature, level of receptor expression, co-receptor, restriction factors, and genetic background. The spread of SARS-CoV-2 also depends on the extent and duration of viral shedding in the infected host as well as population density and behaviour (group living and grooming). Overall, current data indicate that the most at-risk interactions between humans and animals for COVID-19 infection are those involving certain mustelids (such as minks and ferrets), rodents (such as hamsters), lagomorphs (especially rabbits), and felines (including cats). Therefore, special attention should be paid to the risk of SARS-CoV-2 infection associated with pets.

Peste Des Petits Ruminants: A First Retrospective Investigation Among Susceptible Animal Species in Qatar

A retrospective study was conducted to investigate the prevalence of peste des petits ruminants (PPR) in Qatar. Three hundred sixty-eight blood, swabs, and organ tissue samples collected between 2009 and 2016 were screened for PPR viral antibodies, antigens and nucleic acids using ELISA-Ab, ELISA-Ag and rRT-PCR, respectively. Fifteen PPR positive samples were subjected to virus isolation using Vero cell lines. 52% (n=192) of the samples were shown positive for PPR reporting first time infection of 52% (n=71) animal species including sheep, goat, deer, gazelle, addax, Oryx, blackbuck, deer, springbuck and waterbuck. Eight PPR virus (PPRV) field isolates demonstrated classical PPRV cytopathic effect (CPE) and shown positive for the virus antigens proving finally virus isolation. Sheep had the highest infection rate (55%) followed by wild ruminants (54%) and goats (47%). History wise, PPR might exist in Qatar before 2009. A systematic investigation is recommended to identify the risk factors associated with exposure of the susceptible animals to PPR infection, to test the susceptibility of the different species to PPR infection, and to describe the molecular entity and the replicative potentiality of the circulating field strains.