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1.
BMC Vet Res ; 18(1): 140, 2022 Apr 18.
Article in English | MEDLINE | ID: covidwho-1808371

ABSTRACT

BACKGROUND: Porcine epidemic diarrhea virus (PEDV) is one of the most important enteric viruses causing diarrhea in pigs. The establishment of a rapid detection method applicable in field conditions will be conducive to early detection of pathogen and implementation of relevant treatment. A novel nucleic acid amplification method, recombinase polymerase amplification (RPA), has been widely used for infectious disease diagnosis. RESULTS: In the present study, a reverse transcription (RT)-RPA assay combined with lateral flow dipstrip (LFD) was established for the visual detection of PEDV by targeting the N gene. The RT-RPA-LFD assay detected as low as 102 copies/µL of PEDV genomic RNA standard. Moreover, the novel RT-RPA-LFD assay did not show cross-reactivity with common swine pathogens, demonstrating high specificity. The performance of the assay for detection of clinical samples was also evaluated. A total number of 86 clinical samples were tested by RT-RPA-LFD and RT-PCR. The detection results of RT-RPA-LFD were compared with those of RT-PCR, with a coincidence rate of 96.5%. CONCLUSION: The newly established RT-RPA-LFD assay in our study had high sensitivity and specificity, with a potential to use in resource-limited areas and countries.


Subject(s)
Porcine epidemic diarrhea virus , Animals , Nucleic Acid Amplification Techniques/methods , Nucleic Acid Amplification Techniques/veterinary , Porcine epidemic diarrhea virus/genetics , Real-Time Polymerase Chain Reaction/methods , Real-Time Polymerase Chain Reaction/veterinary , Recombinases/genetics , Reverse Transcription , Sensitivity and Specificity , Swine
2.
PLoS Pathog ; 17(11): e1009952, 2021 11.
Article in English | MEDLINE | ID: covidwho-1593026

ABSTRACT

The breadth of animal hosts that are susceptible to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and may serve as reservoirs for continued viral transmission are not known entirely. In August 2020, an outbreak of SARS-CoV-2 occurred on five mink farms in Utah and was associated with high mink mortality (35-55% of adult mink) and rapid viral transmission between animals. The premise and clinical disease information, pathology, molecular characterization, and tissue distribution of virus within infected mink during the early phase of the outbreak are provided. Infection spread rapidly between independently housed animals and farms, and caused severe respiratory disease and death. Disease indicators were most notably sudden death, anorexia, and increased respiratory effort. Gross pathology examination revealed severe pulmonary congestion and edema. Microscopically there was pulmonary edema with moderate vasculitis, perivasculitis, and fibrinous interstitial pneumonia. Reverse transcriptase polymerase chain reaction (RT-PCR) of tissues collected at necropsy demonstrated the presence of SARS-CoV-2 viral RNA in multiple organs including nasal turbinates, lung, tracheobronchial lymph node, epithelial surfaces, and others. Localization of viral RNA by in situ hybridization revealed a more localized infection, particularly of the upper respiratory tract. Whole genome sequencing from multiple mink was consistent with published SARS-CoV-2 genomes with few polymorphisms. The Utah mink SARS-CoV-2 strains fell into Clade GH, which is unique among mink and other animal strains sequenced to date. While sharing the N501T mutation which is common in mink, the Utah strains did not share other spike RBD mutations Y453F and F486L found in nearly all mink from the United States. Mink in the outbreak reported herein had high levels of SARS-CoV-2 in the upper respiratory tract associated with symptomatic respiratory disease and death.


Subject(s)
COVID-19/veterinary , Mink/virology , Animals , COVID-19/epidemiology , COVID-19/mortality , COVID-19/pathology , Disease Outbreaks/veterinary , Farms , Female , Lung/pathology , Male , RNA, Viral/blood , Real-Time Polymerase Chain Reaction/veterinary , SARS-CoV-2/classification , Utah/epidemiology
3.
Transbound Emerg Dis ; 69(4): e823-e830, 2022 Jul.
Article in English | MEDLINE | ID: covidwho-1488270

ABSTRACT

Since the start of the coronavirus disease of 2019 (COVID-19) pandemic, several episodes of human-to-animal severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) transmission have been described in different countries. The role of pets, especially domestic dogs, in the COVID-19 epidemiology is highly questionable and needs further investigation. In this study, we report a case of COVID-19 in a French dog living in close contact with its owners who were COVID-19 patients. The dog presented rhinitis and was sampled 1 week after its owners (a man and a woman) were tested positive for COVID-19. The nasal swabs for the dog tested remained positive for SARS-CoV-2 by reverse transcription quantitative real-time PCR (RT-qPCR) 1 month following the first diagnosis. Specific anti-SARS-CoV-2 antibodies were detectable 12 days after the first diagnosis and persisted for at least 5 months as tested using enzyme-linked immunoassay (ELISA) and automated western blotting. The whole-genome sequences from the dog and its owners were 99%-100% identical (with the man and the woman's sequences, respectively) and matched the B.1.160 variant of concern (Marseille-4 variant), the most widespread in France at the time the dog was infected. This study documents the first detection of B.1.160 in pets (a dog) in France, and the first canine genome recovery of the B.1.160 variant of global concern. Moreover, given the enhanced infectivity and transmissibility of the Marseille-4 variant for humans, this case also highlights the risk that pets may potentially play a significant role in SARS-CoV-2 outbreaks and may transmit the infection to humans. We have evidence of human-to-dog transmission of the Marseille-4 variant since the owners were first to be infected. Finally, owners and veterinarians must be vigilent for canine COVID-19 when dogs are presented with respiratory clinical signs.


Subject(s)
COVID-19 , Dog Diseases , Animals , Antibodies, Viral , COVID-19/diagnosis , COVID-19/epidemiology , COVID-19/veterinary , Dog Diseases/diagnosis , Dog Diseases/epidemiology , Dogs , Female , Humans , Pandemics/veterinary , Real-Time Polymerase Chain Reaction/veterinary , SARS-CoV-2/genetics
4.
J Vet Sci ; 22(6): e70, 2021 Nov.
Article in English | MEDLINE | ID: covidwho-1485292

ABSTRACT

Bats are an important reservoir of several zoonotic diseases. However, the circulation of bat coronaviruses (BatCoV) in live animal markets in Indonesia has not been reported. Genetic characterization of BatCoV was performed by sequencing partial RdRp genes. Real-time polymerase chain reaction based on nucleocapsid protein (N) gene and Enzyme-linked immunosorbent assay against the N protein were conducted to detect the presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral RNA and antibody, respectively. We identified the presence of BatCoV on Cynopterus brachyotis, Macroglossus minimus, and Rousettus amplexicaudatus. The results showed that the BatCoV included in this study are from an unclassified coronavirus group. Notably, SARS-CoV-2 viral RNA and antibodies were not detected in the sampled bats.


Subject(s)
Chiroptera/virology , Coronavirus/classification , Coronavirus/isolation & purification , Animals , Coronavirus/genetics , DNA, Viral/genetics , Enzyme-Linked Immunosorbent Assay/veterinary , Indonesia , Nucleocapsid Proteins/genetics , Real-Time Polymerase Chain Reaction/veterinary , Reverse Transcriptase Polymerase Chain Reaction/veterinary , Species Specificity
5.
Vet Microbiol ; 262: 109243, 2021 Nov.
Article in English | MEDLINE | ID: covidwho-1415830

ABSTRACT

SARS-CoV-2 infects several animal species and SARS-CoV-2 variants of concern (VOCs) may even show (as in humans) enhanced inter- and intra-species transmission rates. We correlated sensitivity data of SARS-CoV-2 rapid antigen tests (RATs) to viral RNA genome equivalents analyzed by real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Further, we checked their suitability for testing animals by assessing saliva and VOC effects. Viral loads up to 2 logs (RNA copy number) under the hypothetical SARS-CoV-2 infectivity threshold were detected by most analyzed RATs. However, while saliva from various animal species showed generally no adverse effects on the RATs' analytical sensitivities, the detection of VOCs B.1.1.7 and B.1.351 was in some RATs inferior to non-VOC viruses.


Subject(s)
Antigens, Viral/isolation & purification , COVID-19 Serological Testing/veterinary , COVID-19/veterinary , Genetic Variation , SARS-CoV-2/isolation & purification , Saliva/virology , Animals , COVID-19/diagnosis , COVID-19/virology , COVID-19 Serological Testing/standards , Chlorocebus aethiops , RNA, Viral/genetics , Real-Time Polymerase Chain Reaction/veterinary , SARS-CoV-2/genetics , SARS-CoV-2/immunology , Vero Cells , Viral Load/veterinary
6.
Vet Res ; 52(1): 121, 2021 Sep 16.
Article in English | MEDLINE | ID: covidwho-1414142

ABSTRACT

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic is causing a global crisis. It is still unresolved. Although many therapies and vaccines are being studied, they are still in their infancy. As this pandemic continues, rapid and accurate research for the development of therapies and vaccines is needed. Therefore, it is necessary to understand characteristics of diseases caused by SARS-CoV-2 through animal models. Syrian hamsters are known to be susceptible to SARS-CoV-2. They were intranasally inoculated with SARS-CoV-2. At 2, 4, 8, 12, and 16 days post-infection (dpi), these hamsters were euthanized, and tissues were collected for ultrastructural and microstructural examinations. Microscopic lesions were prominent in the upper and lower respiratory tracts from 2 and 4 dpi groups, respectively. The respiratory epithelium in the trachea, bronchiole, and alveolar showed pathological changes. Inflammatory cells including neutrophils, lymphocytes, macrophages, and eosinophils were infiltrated in/around tracheal lamina propria, pulmonary vessels, alveoli, and bronchiole. In pulmonary lesions, alveolar wall was thickened with infiltrated inflammatory cells, mainly neutrophils and macrophages. In the trachea, epithelial damages started from 2 dpi and recovered from 8 dpi, consistent with microscopic results, High levels of SARS-CoV-2 nucleoprotein were detected at 2 dpi and 4 dpi. In the lung, lesions were most severe at 8 dpi. Meanwhile, high levels of SARS-CoV-2 were detected at 4 dpi. Electron microscopic examinations revealed cellular changes in the trachea epithelium and alveolar epithelium such as vacuolation, sparse micro-organelle, and poor cellular margin. In the trachea epithelium, the number of cytoplasmic organelles was diminished, and small vesicles were prominent from 2 dpi. Some of these electron-lucent vesicles were filled with virion particles. From 8 dpi, the trachea epithelium started to recover. Because of shrunken nucleus and swollen cytoplasm, the N/C ratio of type 2 pneumocyte decreased at 8 and 12 dpi. From 8 dpi, lamellar bodies on type 2 pneumocyte cytoplasm were increasingly observed. Their number then decreased from 16 dpi. However, there was no significant change in type 1 pneumocyte. Viral vesicles were only observed in the cytoplasm of type 2 pneumocyte. In conclusion, ultra- and micro-structural changes presented in this study may provide useful information for SARS-CoV-2 studies in various fields.


Subject(s)
COVID-19/pathology , Respiratory System/pathology , SARS-CoV-2/pathogenicity , Animals , Cricetinae , Immunohistochemistry/veterinary , Male , Mesocricetus , Pilot Projects , RNA, Viral/chemistry , RNA, Viral/isolation & purification , Real-Time Polymerase Chain Reaction/veterinary , Respiratory System/chemistry , Respiratory System/ultrastructure , Respiratory System/virology , Time Factors , Trachea/pathology , Trachea/ultrastructure , Trachea/virology , Weight Loss
7.
Res Vet Sci ; 140: 229-232, 2021 Nov.
Article in English | MEDLINE | ID: covidwho-1401839

ABSTRACT

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.


Subject(s)
COVID-19 , Cat Diseases , Animals , COVID-19/veterinary , Cat Diseases/diagnosis , Cats , Germany , Humans , Real-Time Polymerase Chain Reaction/veterinary , SARS-CoV-2
8.
Viruses ; 13(9)2021 09 03.
Article in English | MEDLINE | ID: covidwho-1390790

ABSTRACT

Despite the probable zoonotic origin of SARS-CoV-2, only limited research efforts have been made to understand the role of companion animals in SARS-CoV-2 epidemiology. According to recent serological prevalence studies, human-to-companion animal transmission is quite frequent, which led us to consider that the risk of SARS-CoV-2 transmission from animal to human, albeit negligible in the present context, may have been underestimated. In this study, we provide the results of a prospective survey that was conducted to evaluate the SARS-CoV-2 isolation rate by qRT-PCR in dogs and cats with different exposure risks and clinical statuses. From April 2020 to April 2021, we analyzed 367 samples and investigated the presence of SARS-CoV-2 RNA using qRT-PCR. Only four animals tested positive, all of them being cats. Three cats were asymptomatic and one presented a coryza-like syndrome. We describe in detail the infection in two cats and the associated clinical characteristics. Importantly, we obtained SARS-CoV-2 genomes from one infected animal and characterized them as Alpha variants. This represents the first identification of the SARS-CoV-2 Alpha variant in an infected animal in France.


Subject(s)
COVID-19/veterinary , Cat Diseases/virology , Dog Diseases/virology , Animals , COVID-19/epidemiology , COVID-19/transmission , COVID-19/virology , Cat Diseases/epidemiology , Cats , Dog Diseases/epidemiology , Dogs , France/epidemiology , Humans , Male , Pets/virology , Prevalence , Prospective Studies , RNA, Viral , Real-Time Polymerase Chain Reaction/veterinary , SARS-CoV-2/genetics , SARS-CoV-2/isolation & purification , Sequence Analysis, RNA , Virus Shedding
9.
J Vet Diagn Invest ; 33(3): 577-581, 2021 May.
Article in English | MEDLINE | ID: covidwho-1271943

ABSTRACT

The H2 subtypes of avian influenza A viruses (avian IAVs) have been circulating in poultry, and they have the potential to infect humans. Therefore, establishing a method to quickly detect this subtype is pivotal. We developed a TaqMan minor groove binder real-time RT-PCR assay that involved probes and primers based on conserved sequences of the matrix and hemagglutinin genes. The detection limit of this assay was as low as one 50% egg infectious dose (EID50)/mL per reaction. This assay is specific, sensitive, and rapid for detecting avian IAV H2 subtypes.


Subject(s)
Influenza A virus/isolation & purification , Influenza in Birds/diagnosis , Real-Time Polymerase Chain Reaction/veterinary , Reverse Transcriptase Polymerase Chain Reaction/veterinary , Animals , Chick Embryo , Chickens , Real-Time Polymerase Chain Reaction/methods , Reverse Transcriptase Polymerase Chain Reaction/methods , Sensitivity and Specificity
10.
J Small Anim Pract ; 62(8): 662-668, 2021 08.
Article in English | MEDLINE | ID: covidwho-1251728

ABSTRACT

OBJECTIVES: The objectives of this study were to determine the prevalence of canine infectious respiratory disease pathogens among asymptomatic client-owned dogs, and to compare the risks of asymptomatic pathogen carriage between client-owned dogs and dogs in an animal shelter. MATERIALS AND METHODS: Pooled tonsillar, conjunctival and nasal cavity swabs from asymptomatic client-owned dogs (n=133) were tested using a real-time polymerase chain reaction canine respiratory panel. Identical samples from asymptomatic dogs in an animal shelter (n=295) were similarly tested for selected pathogens. Risk differences were calculated between client-owned dogs and shelter dogs for each of the respiratory pathogens included in the analyses. RESULTS: A total of 15 of 133 (11.3%) asymptomatic client-owned dogs were positive for at least one pathogen in the complex. Seven dogs (6.1%) were positive for M. cynos, six (5.2%) were positive for B. bronchiseptica, two (1.7%) were positive for canine herpesvirus type 1 and two (1.7%) were positive for canine respiratory coronavirus. For all eight pathogens tested in both groups, the proportion of positive cases was higher among shelter dogs than among client-owned dogs. Shelter dogs had a higher risk for M. cynos (0.18, 95% confidence interval: 0.12 to 0.25), canine respiratory coronavirus (0.15, 95% confidence interval: 0.10 to 0.19), canine distemper virus (0.06, 95% confidence interval: 0.03 to 0.09), and canine pneumovirus (0.05, 95% confidence interval: 0.03 to 0.08) than client-owned dogs. Odds ratios for M. cynos (0.31, 95% confidence interval: 0.08 to 0.92) and canine respiratory coronavirus (0.05, 95% confidence interval: 0.01 to 0.18) were significantly different between client-owned and shelter dogs. In all cases except for canine herpesvirus type 1, dogs within the shelter population were observed to be at higher risk of exhibiting asymptomatic carriage of a respiratory pathogen as compared to client-owned dogs. The strength of this association was strongest for M. cynos and canine respiratory coronavirus. CLINICAL SIGNIFICANCE: The risk of canine infectious respiratory disease pathogen exposure posed by asymptomatic client-owned dogs is poorly defined. This study also corroborates previous reports of high canine infectious respiratory disease prevalence among clinically healthy shelter dogs, and further determined that the overall prevalence of canine infectious respiratory disease pathogen carriage among clinically healthy client-owned dogs is low but is highest for the traditional pathogen B. bronchiseptica and the emerging pathogen M. cynos.


Subject(s)
Communicable Diseases , Dog Diseases , Respiratory Tract Infections , Animals , Communicable Diseases/veterinary , Dog Diseases/epidemiology , Dogs , Prevalence , Real-Time Polymerase Chain Reaction/veterinary , Respiratory Tract Infections/epidemiology , Respiratory Tract Infections/veterinary
11.
J Vet Diagn Invest ; 33(2): 331-335, 2021 Mar.
Article in English | MEDLINE | ID: covidwho-1058184

ABSTRACT

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of coronavirus disease 2019 (COVID-19), which is an ongoing global health concern. The exact source of the virus has not been identified, but it is believed that this novel coronavirus originated in animals; bats in particular have been implicated as the primary reservoir of the virus. SARS-CoV-2 can also be transmitted from humans to other animals, including tigers, cats, and mink. Consequently, infected people who work directly with bats could transfer the virus to a wild North American bat, resulting in a new natural reservoir for the virus, and lead to new outbreaks of human disease. We evaluated a reverse-transcription real-time PCR panel for detection of SARS-CoV-2 in bat guano. We found the panel to be highly specific for SARS-CoV-2, and able to detect the virus in bat guano samples spiked with SARS-CoV-2 viral RNA. Our panel could be utilized by wildlife agencies to test bats in rehabilitation facilities prior to their release to the wild, minimizing the risk of spreading this virus to wild bat populations.


Subject(s)
COVID-19/transmission , Chiroptera/virology , SARS-CoV-2/isolation & purification , Animals , Animals, Wild , Feces/virology , Humans , Real-Time Polymerase Chain Reaction/veterinary , Sensitivity and Specificity , Zoonoses
12.
J Vet Diagn Invest ; 33(1): 80-86, 2021 Jan.
Article in English | MEDLINE | ID: covidwho-920981

ABSTRACT

In the United States, horses are used for a variety of purposes including recreation, exhibition, and racing. As farm, performance, and companion animals, horses are a unique species from a zoonotic disease risk perspective, and the risks of subclinical infections spreading among horses can pose challenges. Using a nanoscale real-time PCR platform, we investigated the prevalence of 14 enteric pathogens, 11 Escherichia coli genes, and 9 respiratory pathogens in fecal samples from 97 apparently healthy horses at a multi-day horse event. In addition, sugar flotation test was performed for fecal parasites. E. coli f17 was commonly detected, prevalent in 59% of horses, followed closely by Streptococcus equi subsp. zooepidemicus (55%). Additional pathogens recognized included betacoronavirus, Campylobacter jejuni, Cryptosporidium sp., E. coli O157, equine adenovirus 1, equine rhinitis B virus, and others. The use of PCR data may overestimate the true prevalence of these pathogens but provides a sensitive overview of common pathogens present in healthy horses. Our results prompt the continued need for practical biosecurity measures at horse shows, both to protect individuals interacting with these horses and to minimize transmission among horses.


Subject(s)
Animal Husbandry , Cryptosporidiosis/epidemiology , Cryptosporidium/isolation & purification , Escherichia coli Infections/veterinary , Escherichia coli/isolation & purification , Horse Diseases/epidemiology , Animals , Cryptosporidium/genetics , Escherichia coli/genetics , Escherichia coli Infections/epidemiology , Female , Horse Diseases/diagnosis , Horses , Male , New York/epidemiology , Population Surveillance , Real-Time Polymerase Chain Reaction/veterinary
13.
Avian Pathol ; 49(1): 21-28, 2020 Feb.
Article in English | MEDLINE | ID: covidwho-822641

ABSTRACT

Since the emergence of low pathogenic avian influenza (LPAI) H9N2 viruses in Morocco in 2016, severe respiratory problems have been encountered in the field. Infectious bronchitis virus (IBV) is often detected together with H9N2, suggesting disease exacerbation in cases of co-infections. This hypothesis was therefore tested and confirmed in laboratory conditions using specific-pathogen-free chickens. Most common field vaccine programmes were then tested to compare their efficacies against these two co-infecting agents. IBV γCoV/chicken/Morocco/I38/2014 (Mor-IT02) and LPAI virus A/chicken/Morocco/SF1/2016 (Mor-H9N2) were thus inoculated to commercial chickens. We showed that vaccination with two heterologous IBV vaccines (H120 at day one and 4/91 at day 14 of age) reduced the severity of clinical signs as well as macroscopic lesions after simultaneous experimental challenge. In addition, LPAI H9N2 vaccination was more efficient at day 7 than at day 1 in limiting disease post simultaneous challenge.RESEARCH HIGHLIGHTS Simultaneous challenge with IBV and AIV H9N2 induced higher pathogenicity in SPF birds than inoculation with IBV or AIV H9N2 alone.Recommended vaccination programme in commercial broilers to counter Mor-IT02 IBV and LPAIV H9N2 simultaneous infections: IB live vaccine H120 (d1), AIV H9N2 inactivated vaccine (d7), IB live vaccine 4-91 (d14).


Subject(s)
Chickens , Coinfection/veterinary , Coronavirus Infections/veterinary , Infectious bronchitis virus , Influenza A Virus, H9N2 Subtype , Influenza in Birds/virology , Animals , Antibodies, Viral/blood , Chick Embryo , Coinfection/prevention & control , Coinfection/virology , Coronavirus Infections/prevention & control , Coronavirus Infections/virology , Influenza in Birds/prevention & control , Lung/pathology , Morocco , Oropharynx/virology , Pilot Projects , Poultry Diseases/prevention & control , Poultry Diseases/virology , RNA, Viral/chemistry , RNA, Viral/isolation & purification , Real-Time Polymerase Chain Reaction/veterinary , Specific Pathogen-Free Organisms , Trachea/pathology , Vaccination/veterinary , Vaccines, Attenuated , Viral Vaccines , Virus Shedding
14.
Emerg Infect Dis ; 26(1): 168-171, 2020 01.
Article in English | MEDLINE | ID: covidwho-829054

ABSTRACT

Influenza D virus (IDV) can potentially cause respiratory diseases in livestock. We isolated a new IDV strain from diseased cattle in Japan; this strain is phylogenetically and antigenically distinguished from the previously described IDVs.


Subject(s)
Cattle Diseases/epidemiology , Orthomyxoviridae Infections/veterinary , Thogotovirus/genetics , Animals , Cattle/virology , Cattle Diseases/virology , Japan/epidemiology , Orthomyxoviridae Infections/epidemiology , Orthomyxoviridae Infections/virology , Phyllachorales , Real-Time Polymerase Chain Reaction/veterinary
15.
Transbound Emerg Dis ; 67(6): 2609-2614, 2020 Nov.
Article in English | MEDLINE | ID: covidwho-197600

ABSTRACT

The Middle East respiratory syndrome coronavirus (MERS-CoV) is an emergent respiratory virus. Dromedary camels are currently the only known reservoir of MERS-CoV and are capable of transmitting the virus within a herd. The role of semen in the transmission of MERS-CoV has never been investigated as yet, to the best of our knowledge. Our goal was to test semen collected from dromedary camels for MERS-CoV. A total of 67 seminal plasma samples from infertile and 13 from fertile dromedary camels were collected. The RNA was extracted from the samples and tested using commercial real-time PCR. Nine out of sixty-seven infertile animals (13.4%) were positive. The obtained PCR products were sequenced using the conserved MERS-CoV-N gene primers. MERS-CoV-RNA detected in seminal plasma was closely related to the lineage B. To the best of our knowledge, this is the first report about the detection of MERS-CoV-RNA in camel's seminal plasma. Regular testing of semen of common male camels' used for insemination should be considered to avoid a possible spread of the virus through semen.


Subject(s)
Camelus , Coronavirus Infections/veterinary , Middle East Respiratory Syndrome Coronavirus/isolation & purification , Semen/virology , Animals , Coronavirus Infections/virology , Male , Real-Time Polymerase Chain Reaction/veterinary , Saudi Arabia
16.
BMC Vet Res ; 16(1): 114, 2020 Apr 15.
Article in English | MEDLINE | ID: covidwho-60426

ABSTRACT

BACKGROUND: As a pestivirus of the Flaviviridae family, bovine viral diarrhea virus (BVDV), has imposed a large burden on animal husbandry worldwide, and such virus can be transmitted mainly through direct contact with other infected animals and probably via aerosols. In the present study, we aimed to develop a real-time RT-PCR method for detection of BVDV-1 in aerosol samples. METHODS: A pair of primers specific for highly conserved regions of the BVDV-1 5'-UTR was designed. The standard curve and sensitivity of the developed assay were assessed based on 10-fold serial dilutions of RNA molecular standard. The specificity of the assay was evaluated with other pestiviruses and infectious bovine viruses. The clinical performance was examined by testing 169 aerosol samples. RESULTS: The results showed that a good linear relationship existed between the standard curve and the concentration of template. The lowest detection limit was 5.2 RNA molecules per reaction. This assay was specific for detection of BVDV-1, and no amplification was found for other pestiviruses such as classical swine fever virus (CSFV), border disease virus (BDV), and common infectious bovine viruses, including BVDV-2, infectious bovine rhinotracheitis virus (IBRV), bovine parainfluenza virus type 3 (BPIV-3), bovine respiratory syncytial virus (BRSV), bovine ephemeral fever virus (BEFV) and bovine coronavirus (BcoV). The assay was highly reproducible with low variation coefficient values (CVs) for intra-assay and inter-assay. A total of 169 aerosol samples collected from six dairy herds were tested using this method. The results showed that the positive detection rate of BVDV-1 was 17.2% (29/169), which was significantly higher compared with the conventional RT-PCR. Additionally, the positive samples (n = 29) detected by real-time RT-PCR were verified by BVDV RPA-LFD, and a concordance rate of 100% was obtained between them. CONCLUSIONS: Taken together, we developed a real-time RT-PCR assay for quantitative analysis of BVDV-1 in aerosol samples, and our finding provided valuable insights into the risk on aerosol transmission of BVDV-1.


Subject(s)
Diarrhea Virus 1, Bovine Viral/isolation & purification , Genotype , Real-Time Polymerase Chain Reaction/veterinary , Reverse Transcriptase Polymerase Chain Reaction/veterinary , 5' Untranslated Regions/genetics , Aerosols , Air Microbiology , Animals , Diarrhea Virus 1, Bovine Viral/classification , Diarrhea Virus 1, Bovine Viral/genetics , Real-Time Polymerase Chain Reaction/methods , Reproducibility of Results , Reverse Transcriptase Polymerase Chain Reaction/methods , Sensitivity and Specificity
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