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1.
Res Vet Sci ; 140: 229-232, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34534904

RESUMO

Several non-variant of concern SARS-CoV-2 infections in pets have been reported as documented in the OIE and GISAID databases and there is only one fully documented case of an alpha variant of concern (VOC)(B.1.1.7) in the United States so far. Here, we describe the first case in a cat infected with the alpha SARS-CoV-2 variant in Germany. A cat suffering from pneumonia was presented to a veterinary practice. The pneumonia was treated symptomatically, but 16 days later the cat was presented again. Since the owner had been tested positive for a SARS-CoV-2 infection in the meantime, swab samples were taken from the cat and analyzed for SARS-CoV-2 specific nucleic acids. The various RT-qPCR analyses and whole-genome sequencing revealed the presence of the SARS-CoV-2 B.1.1.7 variant in this cat. This study shows that pets living in close contact with SARS-CoV-2 B.1.1.7 infected owners can contract this virus and also suffer from a respiratory disease. It is not clear yet whether onward transmissions to other cats and humans can occur. To minimize transmission risks, pet owners and veterinarians should comply to the hygienic rules published by OIE and others. It must be stated, that infections of cats with SARS-CoV-2 is still a rare event. Cats with clinical signs of a respiratory disease should be presented to a veterinarian, who will decide on further steps.

2.
Viruses ; 13(8)2021 08 19.
Artigo em Inglês | MEDLINE | ID: mdl-34452513

RESUMO

The recent emergence of SARS-CoV-2 in humans from a yet unidentified animal reservoir and the capacity of the virus to naturally infect pets, farmed animals and potentially wild animals has highlighted the need for serological surveillance tools. In this study, the luciferase immunoprecipitation systems (LIPS), employing the spike (S) and nucleocapsid proteins (N) of SARS-CoV-2, was used to examine the suitability of the assay for antibody detection in different animal species. Sera from SARS-CoV-2 naturally-infected mink (n = 77), SARS-CoV-2 experimentally-infected ferrets, fruit bats and hamsters and a rabbit vaccinated with a purified spike protein were examined for antibodies using the SARS-CoV-2 N and/or S proteins. From comparison with the known neutralization status of the serum samples, statistical analyses including calculation of the Spearman rank-order-correlation coefficient and Cohen's kappa agreement were used to interpret the antibody results and diagnostic performance. The LIPS immunoassay robustly detected the presence of viral antibodies in naturally infected SARS-CoV-2 mink, experimentally infected ferrets, fruit bats and hamsters as well as in an immunized rabbit. For the SARS-CoV-2-LIPS-S assay, there was a good level of discrimination between the positive and negative samples for each of the five species tested with 100% agreement with the virus neutralization results. In contrast, the SARS-CoV-2-LIPS-N assay did not consistently differentiate between SARS-CoV-2 positive and negative sera. This study demonstrates the suitability of the SARS-CoV-2-LIPS-S assay for the sero-surveillance of SARS-CoV-2 infection in a range of animal species.


Assuntos
Anticorpos Antivirais/sangue , COVID-19/veterinária , Vison/imunologia , SARS-CoV-2/imunologia , Animais , COVID-19/diagnóstico , COVID-19/epidemiologia , COVID-19/imunologia , Teste Sorológico para COVID-19 , Quirópteros/imunologia , Proteínas do Nucleocapsídeo de Coronavírus/imunologia , Monitoramento Epidemiológico , Furões/imunologia , Imunoprecipitação , Mesocricetus/imunologia , Fosfoproteínas/imunologia , Coelhos/imunologia , Estudos Soroepidemiológicos , Glicoproteína da Espícula de Coronavírus/imunologia
3.
Adv Virus Res ; 110: 59-102, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34353482

RESUMO

Within only one year after the first detection of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), nearly 100 million infections were reported in the human population globally, with more than two million fatal cases. While SARS-CoV-2 most likely originated from a natural wildlife reservoir, neither the immediate viral precursor nor the reservoir or intermediate hosts have been identified conclusively. Due to its zoonotic origin, SARS-CoV-2 may also be relevant to animals. Thus, to evaluate the host range of the virus and to assess the risk to act as potential animal reservoir, a large number of different animal species were experimentally infected with SARS-CoV-2 or monitored in the field in the last months. In this review, we provide an update on studies describing permissive and resistant animal species. Using a scoring system based on viral genome detection subsequent to SARS-CoV-2 inoculation, seroconversion, the development of clinical signs and transmission to conspecifics or humans, the susceptibility of diverse animal species was classified on a semi-quantitative scale. While major livestock species such as pigs, cattle and poultry are mostly resistant, companion animals appear moderately susceptible, while several model animal species used in research, including several Cricetidae species and non-human primates, are highly susceptible to SARS-CoV-2 infection. By natural infections, it became obvious that American minks (Neovison vison) in fur farms, e.g., in the Netherlands and Denmark are highly susceptible resulting in local epidemics in these animals.


Assuntos
COVID-19/veterinária , SARS-CoV-2/fisiologia , Animais , Animais Selvagens/virologia , COVID-19/diagnóstico , COVID-19/transmissão , COVID-19/virologia , Reservatórios de Doenças/veterinária , Reservatórios de Doenças/virologia , Suscetibilidade a Doenças/diagnóstico , Suscetibilidade a Doenças/veterinária , Suscetibilidade a Doenças/virologia , Especificidade de Hospedeiro , Gado/virologia , Modelos Animais , Animais de Estimação/virologia , SARS-CoV-2/isolamento & purificação
4.
Vet Ital ; 57(1): 13-17, 2021 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-34313094

RESUMO

Schmallenberg virus (SBV), a novel Orthobunyavirus, emerged in European domestic ruminants in 2011 causing abortions and malformations in newborns and none or mild clinical symptoms in adult animals. Here, a total of 364 bovine, ovine and caprine serum samples were collected in Kosovo and Albania between May 2014 and August 2016 and analyzed for the presence of anti­SBV antibodies. Sera were tested using an enzyme­linked immunosorbent assay (ELISA), and 48 ELISA­positive samples were subsequently analyzed by serum neutralization test (SNT). The overall percentage of ELISA positive results was 17.9%; 23.1% (53/229) was the prevalence observed in Kosovo (cattle 45.5%, sheep 19.2% and goat 6.8%), while 8.9% (12/135) was that observed in Albania (cattle 11.1%, sheep 0% and goat 20.0%). SNT confirmed the presence of neutralizing antibodies against SBV in all samples tested. This is the first study reporting SBV circulation in domestic ruminants in Kosovo and Albania, with indication that this virus has been present in Kosovo and Albania at least since 2014 without being detected.


Assuntos
Infecções por Bunyaviridae/veterinária , Doenças dos Bovinos/epidemiologia , Doenças das Cabras/epidemiologia , Orthobunyavirus/isolamento & purificação , Doenças dos Ovinos/epidemiologia , Albânia/epidemiologia , Animais , Anticorpos Antivirais/sangue , Infecções por Bunyaviridae/epidemiologia , Bovinos , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/virologia , Doenças das Cabras/diagnóstico , Doenças das Cabras/virologia , Cabras , Kosovo/epidemiologia , Orthobunyavirus/imunologia , Estudos Soroepidemiológicos , Ovinos , Doenças dos Ovinos/diagnóstico , Doenças dos Ovinos/virologia
5.
BMC Vet Res ; 17(1): 242, 2021 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-34247601

RESUMO

BACKGROUND: As a global ruminant pathogen, bovine viral diarrhea virus (BVDV) is responsible for the disease Bovine Viral Diarrhea with a variety of clinical presentations and severe economic losses worldwide. Classified within the Pestivirus genus, the species Pestivirus A and B (syn. BVDV-1, BVDV-2) are genetically differentiated into 21 BVDV-1 and four BVDV-2 subtypes. Commonly, the 5' untranslated region and the Npro protein are utilized for subtyping. However, the genetic variability of BVDV leads to limitations in former studies analyzing genome fragments in comparison to a full-genome evaluation. RESULTS: To enable rapid and accessible whole-genome sequencing of both BVDV-1 and BVDV-2 strains, nanopore sequencing of twelve representative BVDV samples was performed on amplicons derived through a tiling PCR procedure. Covering a multitude of subtypes (1b, 1d, 1f, 2a, 2c), sample matrices (plasma, EDTA blood and ear notch), viral loads (Cq-values 19-32) and species (cattle and sheep), ten of the twelve samples produced whole genomes, with two low titre samples presenting 96 % genome coverage. CONCLUSIONS: Further phylogenetic analysis of the novel sequences emphasizes the necessity of whole-genome sequencing to identify novel strains and supplement lacking sequence information in public repositories. The proposed amplicon-based sequencing protocol allows rapid, inexpensive and accessible obtainment of complete BVDV genomes.

6.
Nat Commun ; 12(1): 4048, 2021 06 30.
Artigo em Inglês | MEDLINE | ID: mdl-34193869

RESUMO

The ongoing SARS-CoV-2 pandemic necessitates the fast development of vaccines. Recently, viral mutants termed variants of concern (VOC) which may escape host immunity have emerged. The efficacy of spike encoding mRNA vaccines (CVnCoV and CV2CoV) against the ancestral strain and the VOC B.1.351 was tested in a K18-hACE2 transgenic mouse model. Naive mice and mice immunized with a formalin-inactivated SARS-CoV-2 preparation were used as controls. mRNA-immunized mice develop elevated SARS-CoV-2 RBD-specific antibody and neutralization titers which are readily detectable, but significantly reduced against VOC B.1.351. The mRNA vaccines fully protect from disease and mortality caused by either viral strain. SARS-CoV-2 remains undetected in swabs, lung, or brain in these groups. Despite lower neutralizing antibody titers compared to the ancestral strain BavPat1, CVnCoV and CV2CoV show complete disease protection against the novel VOC B.1.351 in our studies.


Assuntos
Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Vacinas contra COVID-19/imunologia , COVID-19/prevenção & controle , SARS-CoV-2/imunologia , Glicoproteína da Espícula de Coronavírus/imunologia , Enzima de Conversão de Angiotensina 2/genética , Enzima de Conversão de Angiotensina 2/imunologia , Animais , Linhagem Celular , Chlorocebus aethiops , Genoma Viral/genética , Humanos , Camundongos , Camundongos Transgênicos , SARS-CoV-2/genética , Células Vero
7.
Emerg Infect Dis ; 27(7): 1974-1976, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34152973

RESUMO

We report a therapy cat in a nursing home in Germany infected with severe acute respiratory syndrome coronavirus 2 during a cluster outbreak in the home residents. Although we confirmed prolonged presence of virus RNA in the asymptomatic cat, genome sequencing showed no further role of the cat in human infections on site.


Assuntos
COVID-19 , SARS-CoV-2 , Animais , Gatos , Surtos de Doenças , Alemanha , Humanos , RNA Viral/genética , Aposentadoria
8.
Viruses ; 13(6)2021 05 27.
Artigo em Inglês | MEDLINE | ID: mdl-34072254

RESUMO

Registered cases of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections in the German human population increased rapidly during the second wave of the SARS-CoV-2 pandemic in winter 2020/21. Since domestic cats are susceptible to SARS-CoV-2, the occurrence of trans-species transmission needs to be monitored. A previous serosurvey during the first wave of the pandemic detected antibodies against SARS-CoV-2 in 0.65% of feline serum samples that were randomly sampled across Germany. In the here-presented follow-up study that was conducted from September 2020 to February 2021, the seroprevalence rose to 1.36% (16/1173). This doubling of the seroprevalence in cats is in line with the rise of reported cases in the human population and indicates a continuous occurrence of trans-species transmission from infected owners to their cats.


Assuntos
Anticorpos Antivirais/sangue , COVID-19/veterinária , Doenças do Gato/epidemiologia , SARS-CoV-2/imunologia , Animais , Animais Domésticos , COVID-19/diagnóstico , COVID-19/epidemiologia , COVID-19/transmissão , Doenças do Gato/diagnóstico , Doenças do Gato/transmissão , Gatos , Alemanha/epidemiologia , Humanos , Prevalência , SARS-CoV-2/isolamento & purificação , Estudos Soroepidemiológicos , Zoonoses/diagnóstico , Zoonoses/epidemiologia , Zoonoses/transmissão
9.
Pathogens ; 10(3)2021 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-33803542

RESUMO

Bovine viral diarrhea (BVD) is one of the most important infectious cattle diseases worldwide. The major source of virus transmission is immunotolerant, persistently infected (PI) calves, which makes them the key target of control programs. In the German federal state of Saxony-Anhalt, a very low prevalence was achieved, with more than 99.8% of the cattle herds being free from PI animals since the year 2013. In 2017, BVD virus was detected in a previously disease-free holding (herd size of ~380 cows, their offspring, and fattening bulls). The purchase of two so-called Trojan cows, i.e., dams pregnant with a PI calf, was identified as the source of infection. The births of the PI animals resulted in transient infections of in-contact dams, accompanied by vertical virus transmission to their fetuses within the critical timeframe for the induction of PI calves. Forty-eight days after the birth of the first PI calf, all animals in close contact with the Trojan cows during their parturition period were blood-sampled and serologically examined by a neutralization test and several commercial ELISAs. The resulting seroprevalence strongly depended on the applied test system. The outbreak could be stopped by the immediate elimination of every newborn PI calf and vaccination, and since 2018, no BVD cases have occurred.

10.
Ticks Tick Borne Dis ; 12(4): 101693, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33690089

RESUMO

Tick-borne encephalitis virus (TBEV) is a vector-borne pathogen that can cause serious neurological symptoms in humans. Across large parts of Eurasia TBEV is found in three traditional subtypes: the European, the Siberian and the Far-eastern subtype. Small mammalian animals play an important role in the transmission cycle as they enable the spread of TBEV among the vector tick population. To assess the impact of TBEV infection on its natural hosts, outbred bank voles (Myodes glareolus) were inoculated with one out of four European TBEV strains. Three of these TBEV strains were recently isolated in Germany. The forth one was the TBEV reference strain Neudörfl. Sampling points at 7, 14, 28, and 56 days post inoculation allowed the characterization of the course of infection. At each time point, six animals per strain were euthanized and eleven organ samples (brain, spine, lung, heart, small and large intestine, liver, spleen, kidney, bladder, sexual organ) as well as whole blood and serum samples were collected. The majority of bank voles (92/96) remained clinically unaffected after the inoculation with TBEV, but still developed a systemic infection during the first week, which transitioned to a viraemia and an infestation of the brain in some animals for the remainder of the first month. Viral RNA was found in whole blood samples of several animals (50/96), but only in a small fraction of the corresponding serum samples (4/50). From the whole blood, virus was successfully reisolated in cell culture until 14 days after inoculation. Less than five percent of all inoculated bank voles (4/96) displayed signs of distress in combination with a rapid weight loss and had to be euthanized prematurely. Overall, the recently isolated TBEV strains showed marked differences, such as a more frequent development of long-term viraemia and a higher detection rate of viral RNA in various organs, in comparison to the reference strain Neudörfl. Overall, our data suggest that the bank vole is a potential amplifying host in the TBEV transmission cycle and appears to be highly adapted to circulating TBEV strains.


Assuntos
Arvicolinae , Vírus da Encefalite Transmitidos por Carrapatos/fisiologia , Encefalite Transmitida por Carrapatos/veterinária , Doenças dos Roedores/virologia , Animais , Encefalite Transmitida por Carrapatos/virologia , Feminino , Alemanha , Masculino
11.
Emerg Infect Dis ; 27(4): 1193-1195, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33754987

RESUMO

After experimental inoculation, severe acute respiratory syndrome coronavirus 2 infection was confirmed in bank voles by seroconversion within 8 days and detection of viral RNA in nasal tissue for up to 21 days. However, transmission to contact animals was not detected. Thus, bank voles are unlikely to establish effective transmission cycles in nature.


Assuntos
Arvicolinae , COVID-19 , Transmissão de Doença Infecciosa , Doenças dos Roedores , Soroconversão , Eliminação de Partículas Virais , Animais , Anticorpos Antivirais , COVID-19/imunologia , COVID-19/transmissão , COVID-19/virologia , Teste Sorológico para COVID-19 , Modelos Animais de Doenças , Suscetibilidade a Doenças , Mucosa Nasal/virologia , Doenças dos Roedores/imunologia , Doenças dos Roedores/transmissão , Doenças dos Roedores/virologia
12.
Transbound Emerg Dis ; 68(5): 2696-2702, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-33527715

RESUMO

African swine fever (ASF) is a highly pathogenic viral disease affecting all Suidae, with Ornithodoros moubata complex soft ticks acting as the biological arthropod vectors of the causative agent, African swine fever virus (ASFV). While ASFV is also transmissible via direct contact, pig products and fomites, other arthropods may be involved in virus transmission and persistence. Therefore, we checked various groups of blood-feeding arthropods collected during summer 2017 in wild boar habitats on the Estonian Island of Saaremaa for the presence of ASFV. Saaremaa had the highest ASF infection prevalences in Estonia in 2017, with an incidence of 9% among hunted wild boar. In addition to ASFV, we tested for other selected pathogens. In total, 784 ticks, 6,274 culicoid biting midges, 77 tabanids and 757 mosquitoes were tested as individuals or pools. No ASFV-DNA was found in any of them although about 20% of the tick samples tested positive for swine DNA. By contrast, tick-borne encephalitis virus-RNA was detected in one out of 37 tick pools (2.7%) and Borrelia burgdorferi s.l.-DNA in 20 individual ticks and 17 tick pools (25.2% of all samples). No Schmallenberg virus was detected in the Culicoides specimens. In conclusion, we found no evidence for Ixodes ricinus ticks, Culicoides punctatus and Obsoletus complex biting midges, Aedes spp., Anopheles spp. and Culiseta annulata mosquitoes, and Haematopota pluvialis tabanids playing a role in ASFV transmission in the wild boar population in Estonia.

13.
PLoS Pathog ; 17(1): e1009247, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33497419

RESUMO

Schmallenberg virus (SBV) is the cause of severe fetal malformations when immunologically naïve pregnant ruminants are infected. In those malformed fetuses, a "hot-spot"-region of high genetic variability within the N-terminal region of the viral envelope protein Gc has been observed previously, and this region co-localizes with a known key immunogenic domain. We studied a series of M-segments of those SBV variants from malformed fetuses with point mutations, insertions or large in-frame deletions of up to 612 nucleotides. Furthermore, a unique cell-culture isolate from a malformed fetus with large in-frame deletions within the M-segment was analyzed. Each Gc-protein with amino acid deletions within the "hot spot" of mutations failed to react with any neutralizing anti-SBV monoclonal antibodies or a domain specific antiserum. In addition, in vitro virus replication of the natural deletion variant could not be markedly reduced by neutralizing monoclonal antibodies or antisera from the field. The large-deletion variant of SBV that could be isolated in cell culture was highly attenuated with an impaired in vivo replication following the inoculation of sheep. In conclusion, the observed amino acid sequence mutations within the N-terminal main immunogenic domain of glycoprotein Gc result in an efficient immune evasion from neutralizing antibodies in the special environment of a developing fetus. These SBV-variants were never detected as circulating viruses, and therefore should be considered to be dead-end virus variants, which are not able to spread further. The observations described here may be transferred to other orthobunyaviruses, particularly those of the Simbu serogroup that have been shown to infect fetuses. Importantly, such mutant strains should not be included in attempts to trace the spatial-temporal evolution of orthobunyaviruses in molecular-epidemiolocal approaches during outbreak investigations.


Assuntos
Anticorpos Antivirais/imunologia , Infecções por Bunyaviridae/veterinária , Doenças dos Bovinos/virologia , Variação Genética , Orthobunyavirus/genética , Doenças dos Ovinos/virologia , Proteínas do Envelope Viral/genética , Animais , Anticorpos Neutralizantes/imunologia , Infecções por Bunyaviridae/virologia , Bovinos , Feminino , Feto , Glicoproteínas/genética , Glicoproteínas/imunologia , Mutação , Orthobunyavirus/imunologia , Orthobunyavirus/fisiologia , RNA Viral/genética , Deleção de Sequência , Ovinos , Proteínas do Envelope Viral/imunologia , Replicação Viral
14.
Transbound Emerg Dis ; 68(3): 1531-1540, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-32910551

RESUMO

Shuni virus (SHUV), an insect-transmitted orthobunyavirus of the Simbu serogroup within the family Peribunyaviridae, may induce severe congenital malformations when naïve ruminants are infected during gestation. Only recently, another clinical presentation in cattle, namely neurological disease after postnatal infection, was reported. To characterize the course of the disease under experimental conditions and to confirm a causal relationship between the virus and the neurological disorders observed in the field, six calves each were experimentally inoculated (subcutaneously) with two different SHUV strains from both clinical presentations, that is encephalitis and congenital malformation, respectively. Subsequently, the animals were monitored clinically, virologically and serologically for three weeks. All animals inoculated with the 'encephalitis strain' SHUV 2162/16 developed viremia for three to four consecutive days, seroconverted, and five out of six animals showed elevated body temperature for up to three days. No further clinical signs such as neurological symptoms were observed in any of these animals. However, four out of six animals developed a non-suppurative meningoencephalitis, characterized by perivascular cuffing and glial nodule formation. Moreover, SHUV genome could be visualized in brain tissues of the infected animals by in situ hybridization. In contrast to the 'encephalitis SHUV strain', in animals subcutaneously inoculated with the strain isolated from a malformed newborn (SHUV 2504/3/14), which expressed a truncated non-structural protein NSs, a major virulence factor, no viremia or seroconversion, was observed, demonstrating an expected severe replication defect of this strain in vivo. The lack of viremia further indicates that virus variants evolving in malformed foetuses may represent attenuated artefacts as has been described for closely related viruses. As the neuropathogenicity of SHUV could be demonstrated under experimental conditions, this virus should be included in differential diagnosis for encephalitis in ruminants, and cattle represent a suitable animal model to study the pathogenesis of SHUV.


Assuntos
Infecções por Bunyaviridae/veterinária , Doenças dos Bovinos/diagnóstico , Meningoencefalite/veterinária , Orthobunyavirus/fisiologia , Animais , Infecções por Bunyaviridae/complicações , Infecções por Bunyaviridae/diagnóstico , Infecções por Bunyaviridae/virologia , Bovinos , Doenças dos Bovinos/virologia , Diagnóstico Diferencial , Modelos Animais de Doenças , Feminino , Masculino , Meningoencefalite/diagnóstico , Meningoencefalite/virologia
15.
Transbound Emerg Dis ; 68(2): 253-257, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32536002

RESUMO

To combat the COVID-19 pandemic, millions of PCR tests are performed worldwide. Any deviation of the diagnostic sensitivity and specificity will reduce the predictive values of the test. Here, we report the occurrence of contaminations of commercial primers/probe sets with the SARS-CoV-2 target sequence of the RT-qPCR as an example for pitfalls during PCR diagnostics affecting diagnostic specificity. In several purchased in-house primers/probe sets, quantification cycle values as low as 17 were measured for negative control samples. However, there were also primers/probe sets that displayed very low-level contaminations, which were detected only during thorough internal validation. Hence, it appears imperative to pre-test each batch of reagents extensively before use in routine diagnosis, to avoid false-positive results and low positive predictive value in low-prevalence situations. As such, contaminations may have happened more widely, and COVID-19 diagnostic results should be re-assessed retrospectively to validate the epidemiological basis for control measures.


Assuntos
Teste para COVID-19/normas , COVID-19/diagnóstico , Pandemias , Reação em Cadeia da Polimerase em Tempo Real/normas , SARS-CoV-2 , Benchmarking , Primers do DNA , Contaminação de Equipamentos , Alemanha , Humanos , Sensibilidade e Especificidade
16.
Transbound Emerg Dis ; 2020 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-33191578

RESUMO

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a pandemic with millions of infected humans and hundreds of thousands of fatalities. As the novel disease - referred to as COVID-19 - unfolded, occasional anthropozoonotic infections of animals by owners or caretakers were reported in dogs, felid species and farmed mink. Further species were shown to be susceptible under experimental conditions. The extent of natural infections of animals, however, is still largely unknown. Serological methods will be useful tools for tracing SARS-CoV-2 infections in animals once test systems are evaluated for use in different species. Here, we developed an indirect multi-species ELISA based on the receptor-binding domain (RBD) of SARS-CoV-2. The newly established ELISA was evaluated using 59 sera of infected or vaccinated animals, including ferrets, raccoon dogs, hamsters, rabbits, chickens, cattle and a cat, and a total of 220 antibody-negative sera of the same animal species. Overall, a diagnostic specificity of 100.0% and sensitivity of 98.31% were achieved, and the functionality with every species included in this study could be demonstrated. Hence, a versatile and reliable ELISA protocol was established that enables high-throughput antibody detection in a broad range of animal species, which may be used for outbreak investigations, to assess the seroprevalence in susceptible species or to screen for reservoir or intermediate hosts.

17.
Vaccines (Basel) ; 8(4)2020 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-33023099

RESUMO

Bovine viral diarrhea virus (BVDV), a pestivirus which exists in the two distinct species BVDV-1 (syn. Pestivirus A) and BVDV-2 (syn. Pestivirus B), is the causative agent of one of the most widespread and economically important virus infections in cattle. For economic as well as for animal health reasons, an increasing number of national BVDV control programs were recently implemented. The main focus lies on the detection and removal of persistently infected cattle. The application of efficient marker or DIVA (differentiation of infected from vaccinated animals) vaccines would be beneficial for the eradication success in regions with a high BVDV prevalence to prevent fetal infection and it would allow serological monitoring of the BVDV status also in vaccinated farms. Therefore, a marker vaccine based on the cytopathic (cp) BVDV-1b strain CP7 was constructed as a synthetic backbone (BVDV-1b_synCP7). For serological discrimination of vaccinated from infected animals, the viral protein Erns was substituted by the heterologous Erns of Bungowannah virus (BuPV, species Pestivirus F). In addition, the vaccines were attenuated by a deletion within the type I interferon inhibitor Npro protein encoding sequence. The BVDV-2 vaccine candidate is based on the genetic sequence of the glycoproteins E1 and E2 of BVDV-2 strain CS8644 (CS), which were introduced into the backbone of BVDV-1b_synCP7_ΔNpro_Erns Bungo in substitution of the homologous glycoproteins. Vaccine virus recovery resulted in infectious cytopathic virus chimera that grew to titers of up to 106 TCID50/mL. Both synthetic chimera BVDV-1b_synCP7_ΔNpro_Erns Bungo and BVDV-1b_synCP7_ΔNpro_Erns Bungo_E1E2 BVDV-2 CS were avirulent in cattle, provided a high level of protection in immunization and challenge experiments against both BVDV species and allowed differentiation of infected from vaccinated cattle. Our study presents the first report on an efficient BVDV-1 and -2 modified live marker vaccine candidate and the accompanying commercially available serological marker ELISA system.

18.
Emerg Infect Dis ; 26(12): 2979-2981, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33034284

RESUMO

We inoculated 6 cattle with severe acute respiratory syndrome coronavirus 2 and kept them together with 3 uninoculated cattle. We observed viral replication and specific seroreactivity in 2 inoculated animals, despite high levels of preexisting antibody titers against a bovine betacoronavirus. The in-contact animals did not become infected.


Assuntos
COVID-19/transmissão , SARS-CoV-2/genética , Animais , Bovinos/virologia , Pandemias , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Zoonoses Virais/transmissão , Replicação Viral
19.
Emerg Infect Dis ; 26(12): 2982-2985, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33089771

RESUMO

Raccoon dogs might have been intermediate hosts for severe acute respiratory syndrome-associated coronavirus in 2002-2004. We demonstrated susceptibility of raccoon dogs to severe acute respiratory syndrome coronavirus 2 infection and transmission to in-contact animals. Infected animals had no signs of illness. Virus replication and tissue lesions occurred in the nasal conchae.


Assuntos
COVID-19/transmissão , SARS-CoV-2/genética , Animais , COVID-19/virologia , Modelos Animais de Doenças , Suscetibilidade a Doenças/virologia , Pandemias , Cães Guaxinins/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Zoonoses Virais , Eliminação de Partículas Virais
20.
Viruses ; 12(8)2020 08 06.
Artigo em Inglês | MEDLINE | ID: mdl-32781607

RESUMO

Bovine viral diarrhea virus (BVDV) entry into a host cell is mediated by the interaction of the viral glycoprotein E2 with the cellular transmembrane CD46 receptor. In this study, we generated a stable Madin-Darby Bovine Kidney (MDBK) CD46-knockout cell line to study the ability of different pestivirus A and B species (BVDV-1 and -2) to escape CD46-dependent cell entry. Four different BVDV-1/2 isolates showed a clearly reduced infection rate after inoculation of the knockout cells. However, after further passaging starting from the remaining virus foci on the knockout cell line, all tested virus isolates were able to escape CD46-dependency and grew despite the lack of the entry receptor. Whole-genome sequencing of the escape-isolates suggests that the genetic basis for the observed shift in infectivity is an amino acid substitution of an uncharged (glycine/asparagine) for a charged amino acid (arginine/lysine) at position 479 in the ERNS in three of the four isolates tested. In the fourth isolate, the exchange of a cysteine at position 441 in the ERNS resulted in a loss of ERNS dimerization that is likely to influence viral cell-to-cell spread. In general, the CD46-knockout cell line is a useful tool to analyze the role of CD46 for pestivirus replication and the virus-receptor interaction.


Assuntos
Vírus da Diarreia Viral Bovina Tipo 1/fisiologia , Vírus da Diarreia Viral Bovina Tipo 2/fisiologia , Proteína Cofatora de Membrana/genética , Receptores Virais/genética , Proteínas do Envelope Viral/genética , Adaptação Biológica , Substituição de Aminoácidos , Animais , Sistemas CRISPR-Cas , Bovinos , Vírus da Diarreia Viral Bovina Tipo 1/genética , Vírus da Diarreia Viral Bovina Tipo 2/genética , Cães , Técnicas de Inativação de Genes , Interações Hospedeiro-Patógeno , Células Madin Darby de Rim Canino , Proteína Cofatora de Membrana/metabolismo , Multimerização Proteica , Receptores Virais/metabolismo , Ribonucleoproteínas/metabolismo , Proteínas do Envelope Viral/química , Proteínas do Envelope Viral/metabolismo , Internalização do Vírus , Replicação Viral
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