The SARS-CoV-2 pandemic has heightened interest in the monitoring and surveillance of coronaviruses in wildlife. Testing for the virus in animals can provide valuable insights into viral reservoirs, transmission, and pathogenesis. In this study, we present the results of the molecular surveillance project focused on coronaviruses in Senegalese wildlife. During the project, we screened fecal samples of the wild animals living in the Bandia Reserve (ten non-human primates, one giraffe, and two white rhinoceros) and the free-living urban population of African four-toed hedgehogs in Ngaparou. The results showed the absence of coronaviruses in hedgehogs, non-human primates, and a giraffe. A single positive sample was obtained from a white rhinoceros. The sequencing results of amplified RdRp gene confirmed that the detected virus was SARS-CoV-2. This study represents the first documented instance of molecular detection of SARS-CoV-2 in white rhinoceros and, therefore, extends our knowledge of possible SARS-CoV-2 hosts.

Coronaviruses (members of the Coronaviridae family) are prominent in veterinary medicine, with several known infectious agents commonly reported. In contrast, human medicine has disregarded coronaviruses for an extended period. Within the past two decades, coronaviruses have caused three major outbreaks. One such outbreak was the coronavirus disease 2019 (COVID-19) caused by the coronavirus severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Over the 3-year COVID-19 outbreak, several instances of zooanthroponosis have been documented, which pose risks for virus modifications and possible re-emergence of the virus into the human population, causing a new epidemic and possible threats for vaccination or treatment failure. Therefore, widespread screening of animals is an essential technique for mitigating future risks and repercussions. However, mass detection of SARS-CoV-2 in wild animals might be challenging. In silico prediction modeling, experimental studies conducted on various animal species, and natural infection episodes recorded in various species might provide information on the potential threats to wildlife. They may be useful for diagnostic and mass screening purposes. In this review, the possible methods of wildlife screening, based on experimental data and environmental elements that might play a crucial role in its effective implementation, are reviewed. Keywords: angiotensin-converting enzyme 2, coronavirus disease 2019 in animals, severe acute respiratory syndrome coronavirus-2, wildlife surveillance.

Coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), first emerged in Wuhan in 2019 and rapidly spread worldwide. During the course of the COVID-19 pandemic, numerous reports highlighted infections of wild animals by SARS-CoV-2. Nevertheless, further research is required to understand the virus potential to infect various animal species, which is crucial for evaluating its future evolution and the potential reemergence of SARS-CoV-2. The total concentration of immunoglobulin G (IgG) represents a valuable yet underused diagnostic parameter for health assessments in wild animals, primarily due to the absence of effective diagnostic tools. A protein A–based indirect ELISA can serve as an efficient method to identify IgG antibodies against different pathogens in wildlife surveillance programs. To develop a multispecies protein A-ELISA for IgG detection against SARS-CoV-2, we used 44 animal species serum samples to ascertain the protein A binding affinity, and 88 serum samples, chosen for the strong binding affinity to protein A, were used to identify IgG antibodies against SARS-CoV-2. The serum samples were obtained from animals housed in Safari Park Dvůr Králové, Czech Republic. The zoo animals are in proximity to humans, facilitating the exploration of potential reverse transmission events of SARS-CoV-2 from humans to animals. Also, they undergo routine veterinary examinations, providing convenient access to blood samples. Therefore, they can be easily used for the development of protein A–based indirect ELISA for wildlife disease surveillance programs. Based on the ELISA results, antibodies to SARS-CoV-2 were detected in the sera of 16 animals. To further confirm these findings, the ELISA-positive samples were subjected to virus neutralization assays. This additional testing revealed the presence of SARS-CoV-2 neutralizing antibodies in the serum of two white rhinoceros (Ceratotherium simum simum) and one Persian leopard (Panthera pardus tulliana).

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) affects both humans and a wide range of mammalian species globally. Between July 2022 and January 2023, fifteen blood samples were collected from twelve different animal species during veterinary examinations, as well as for health control at Wilhelma Zoo, Germany. These samples were later analyzed for the presence of SARS-CoV-2 antibodies. The serum analysis from two gorillas indicated the presence of antibodies specific to the nucleocapsid protein of SARS-CoV-2, suggesting previous infection. These gorillas were sampled in August and September 2022, during which time they exhibited symptoms such as apathy, anorexia, vomiting, and moderate diarrhea—symptoms not typically associated with COVID-19. Given that several periods of other unusual signs have been observed in the gorillas kept in Wilhelma Zoo since the onset of the COVID-19 pandemic, it remains uncertain whether these symptoms were directly related to SARS-CoV-2 infection or if these gorillas underwent clinically inapparent infection before. Nonetheless, this study underscores the importance of ongoing animal screening in zoos to better understand the spread of SARS-CoV-2 among different animal species.