The mutual dependence of human and animal health is central to the One Health initiative as an integrated strategy for infectious disease control and management. A crucial element of the One Health includes preparation and response to influenza A virus (IAV) threats at the human-animal interface. The IAVs are characterized by extensive genetic variability, they circulate among different hosts and can establish host-specific lineages. The four main hosts are: avian, swine, human and equine, with occasional transmission to other mammalian species. The host diversity is mirrored in the range of the RT-qPCR assays for IAV detection. Different assays are recommended by the responsible health authorities for generic IAV detection in birds, swine or humans. In order to unify IAV monitoring in different hosts and apply the One Health approach, we developed a single RT-qPCR assay for universal detection of all IAVs of all subtypes, species origin and global distribution. The assay design was centred on a highly conserved region of the IAV matrix protein (MP)-segment identified by a comprehensive analysis of 99,353 sequences. The reaction parameters were effectively optimised with efficiency of 93-97% and LOD95% of approximately ten IAV templates per reaction. The assay showed high repeatability, reproducibility and robustness. The extensive in silico evaluation demonstrated high inclusivity, i.e. perfect sequence match in the primers and probe binding regions, established as 94.6% for swine, 98.2% for avian and 100% for human H3N2, pandemic H1N1, as well as other IAV strains, resulting in an overall predicted detection rate of 99% on the analysed dataset. The theoretical predictions were confirmed and extensively validated by collaboration between six veterinary or human diagnostic laboratories on a total of 1970 specimens, of which 1455 were clinical and included a diverse panel of IAV strains.

• MeSH
• chřipka lidská diagnóza   virologie   MeSH
• infekce viry z čeledi Orthomyxoviridae diagnóza   virologie   MeSH
• lidé MeSH
• nemoci prasat diagnóza   virologie   MeSH
• One Health MeSH
• polymerázová řetězová reakce s reverzní transkripcí metody   MeSH
• prasata MeSH
• ptačí chřipka u ptáků diagnóza   virologie   MeSH
• ptáci virologie   MeSH
• reprodukovatelnost výsledků MeSH
• virus chřipky A, podtyp H1N1 genetika   izolace a purifikace   MeSH
• virus chřipky A, podtyp H3N2 genetika   izolace a purifikace   MeSH
• virus chřipky A genetika   izolace a purifikace   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH

Porcine reproductive and respiratory syndrome virus (PRRSV) predisposes pigs to secondary bacterial infection caused by Haemophilus parasuis. The aim of the present study was to analyse the immune response of monocyte-derived macrophages (MDMs), serving as a model of macrophages accumulating at the site of inflammation. The second part of the study was focused on the role of IFNα in the production of inflammatory cytokines in co-infected MDMs. Concurrent infection with PRRSV and H. parasuis decreased gene expression of pro-inflammatory cytokines (IL-1β, IL-8) in MDMs in comparison with MDMs infected with PRRSV or H. parasuis alone. Our data showed that MDMs express IFNα after PRRSV infection. Thereafter, we exposed cells to the experimental addition of IFNα and a subsequent infection with H. parasuis, and detected a decreased expression/production of pro-inflammatory cytokines. Thus, we assume that IFNα, produced after PRRSV infection, could affect the immune response of monocyte-derived macrophages. Down-regulation of pro-inflammatory cytokine expression in inflammatory macrophages may allow the development of secondary bacterial infections in pigs.

• MeSH
• bronchoalveolární laváž MeSH
• cytokiny genetika   imunologie   MeSH
• Haemophilus parasuis imunologie   patogenita   MeSH
• hemofilové infekce mikrobiologie   veterinární   MeSH
• interferon alfa imunologie   farmakologie   MeSH
• makrofágy účinky léků   imunologie   mikrobiologie   virologie   MeSH
• nemoci prasat mikrobiologie   virologie   MeSH
• prasata MeSH
• reprodukční a respirační syndrom prasat virologie   MeSH
• viabilita buněk MeSH
• virus reprodukčního a respiračního syndromu prasat imunologie   patogenita   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

To determine whether hepatitis E virus (HEV) was distributed in the population of wild boars in South Moravia between 1990 and 2008, a total of 366 samples of archived sera from wild boars were investigated using serological (commercial ELISA) and molecular (RT-qPCR) methods. A total of 31 (8.5%) wild boars were seropositive, and from two of them, RNA sequences were recovered by nested RT-PCR. The presented results, with one of the oldest animal's HEV-positive serum (collected in 1990), suggest that wild boars may be a reservoir of HEV in the Czech Republic and that this virus has been circulating in studied areas for more than 20 years.

• MeSH
• ELISA MeSH
• hepatitida E krev   epidemiologie   veterinární   MeSH
• nemoci prasat epidemiologie   virologie   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• prasata MeSH
• Sus scrofa virologie   MeSH
• virus hepatitidy E genetika   izolace a purifikace   MeSH
• zdroje nemoci MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

Porcine cytomegalovirus (PCMV) causes lifelong latent infections in swine. The pathogen is occasionally associated with inclusion body rhinitis and pneumonia in piglets, reproductive disorders in pregnant sows and respiratory disease complex in older pigs. Immunosuppressive potential of PCMV infection is discussed. Macrophages were recognised as one of target cell types where propagation of virus occurs. The aim of present study was to set up model PCMV infection of monocyte derived macrophages (MDMs) in vitro for PCMV immunobiology research. Obtained results showed that PCMV is able to infect and propagate in MDMs. Possible immunosuppressive effect of PCMV on infected macrophages was evaluated by measurement of immune relevant gene expression in MDMs. Infection decreased expression of IL-8 and TNF-α (pro-inflammatory cytokines) and increased expression of IL-10 (anti-inflammatory cytokine) on mRNA transcription level. Obtained data support hypothesis that higher sensitivity of animals to coinfection with other swine pathogens and its more severe clinical manifestations could potentially be the consequence of PCMV infection.

• MeSH
• cytokiny imunologie   MeSH
• cytomegalovirové infekce imunologie   veterinární   MeSH
• Cytomegalovirus fyziologie   MeSH
• exprese genu imunologie   MeSH
• interleukin-10 imunologie   MeSH
• interleukin-8 imunologie   MeSH
• makrofágy imunologie   virologie   MeSH
• nemoci prasat imunologie   virologie   MeSH
• prasata MeSH
• přirozená imunita MeSH
• TNF-alfa imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Porcine deltacoronavirus (PDCoV) is a recently discovered RNA virus that belongs to the family Coronaviridae and genus Deltacoronavirus. This virus causes enteric disease in piglets that is characterized by enteritis and diarrhoea. In our present investigation, 189 diarrhoeic samples were collected between July 2016 and May 2017 from Tibetan pigs inhabiting in three different provinces surrounding the Qinghai-Tibet Plateau of China. We then applied the molecular-based method of reverse transcription polymerase chain reactions (RT-PCRs) to detect the presence of PDCoV in collected samples, and RT-PCR indicated that the prevalence of PDCoV was 3.70% (7/189) in Tibetan pigs. Four of 7 PDCoV-positive pigs were monoinfections of PDCoV, three samples were co-infections of PDCoV with porcine epidemic diarrhoea virus (PEDV), and 52 (27.51%) samples were positive for PEDV. Four strains with different full-length genomes were identified (CHN/GS/2016/1, CHN/GS/2016/2, CHN/GS-/2017/1 and CHN/QH/2017/1), and their genomes were used to analyse the characteristics of PDCoV currently prevalent in Tibetan pigs. We found a 3-nt insertion in the spike gene in four strains in Tibetan pigs. Phylogenetic analysis of the complete genome and spike and nucleocapsid gene sequences revealed that these strains shared ancestors with the strain CHN-AH-2004, which was found in pigs from the Anhui province of China mainland. However, PDCoV strains from Tibetan pigs formed different branches within the same cluster, implying continuous evolution in the field. Our present findings highlight the importance of epidemiologic surveillance to limit the spread of PDCoV in livestock at high altitudes in China.

• MeSH
• Coronavirus genetika   izolace a purifikace   MeSH
• feces virologie   MeSH
• fylogeneze MeSH
• genom virový genetika   MeSH
• koronavirové infekce diagnóza   epidemiologie   veterinární   virologie   MeSH
• kvantitativní polymerázová řetězová reakce veterinární   MeSH
• nemoci prasat diagnóza   epidemiologie   virologie   MeSH
• prasata virologie   MeSH
• prevalence MeSH
• průjem veterinární   virologie   MeSH
• sekvenční analýza DNA MeSH
• virové proteiny genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Tibet epidemiologie   MeSH

During a severe outbreak of diarrhoea and vomiting in a pig herd in Central Eastern Europe, faecal samples were tested positive for porcine epidemic diarrhoea virus (PEDV) and negative for transmissible gastroenteritis virus (TGEV) using a commercial RT-qPCR assay that can detect both of these coronaviruses. However, further analyses, using other TGEV- and PEDV-specific RT-qPCR assays, provided results inconsistent with infection by either of these viruses. Sequencing of an amplicon (ca. 1.6 kb), generated by an RT-PCR specific for the PEDV S-gene, indicated a very close similarity (ca. 99% identity) to recently described chimeric viruses termed swine enteric coronaviruses (SeCoVs). These viruses (with an RNA genome of ca. 28 kb) were first identified in Italy in samples from 2009 but have not been detected there since 2012. A closely related virus was detected in archived samples in Germany from 2012, but has not been detected subsequently. Building on the initial sequence data, further amplicons were generated and over 9 kb of sequence corresponding to the 3'-terminus of the new SeCoV genome was determined. Sequence comparisons showed that the three known SeCoVs are ≥98% identical across this region and contain the S-gene and 3a sequences from PEDV within a backbone of TGEV, but the viruses are clearly distinct from each other. It is demonstrated, for the first time, that pigs from within the SeCoV-infected herd seroconverted against PEDV but tested negative in a TGEV-specific ELISA that detects antibodies against the S protein. These results indicate that SeCoV is continuing to circulate in Europe and suggest it can cause a disease that is very similar to PED. Specific detection of the chimeric SeCoVs either requires development of a new diagnostic RT-qPCR assay or the combined use of assays targeting the PEDV S-gene and another part of the TGEV genome.

• MeSH
• ELISA MeSH
• feces virologie   MeSH
• koronavirové infekce veterinární   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• molekulární sekvence - údaje MeSH
• nemoci prasat virologie   MeSH
• prasata MeSH
• přenosná gastroenteritida prasat diagnóza   virologie   MeSH
• Sus scrofa MeSH
• virus epidemického průjmu prasat genetika   izolace a purifikace   MeSH
• virus přenosné gastroenteritidy prasat genetika   izolace a purifikace   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Evropa MeSH
• Itálie MeSH
• Německo MeSH
• východní Evropa MeSH

Population of wild boar is increasing in the whole Europe, the animals migrate close to human habitats which greatly increases the possibility of natural transmission between domestic animals or humans and wild boars. The aim of the study was to estimate in population of free-living wild boar in the Czech Republic the prevalence of enteric viral pathogens, namely rotavirus groups A and C (RVA and RVC), porcine reproductive and respiratory syndrome virus (PRRSV), and members of family Coronaviridae (transmissible gastroenteritis virus - TGEV, porcine epidemic diarrhea virus - PEDV, porcine respiratory coronavirus - PRCV, and porcine hemagglutination encephalomyelitis virus - PHEV) and Picornaviridae,(teschovirus A - PTV, sapelovirus A - PSV, and enterovirus G - EV-G). In our study, stool samples from 203 wild boars culled during hunting season 2014-2015 (from October to January) were examined by RT-PCR. RVA was detected in 2.5% of tested samples. Nucleotide analysis of VP7, VP4, and VP6 genes revealed that four RVA strains belong to G4P[25]I1, G4P[6]I5, G11P[13]I5, and G5P[13]I5 genotypes and phylogenetic analysis suggested close relation to porcine and human RVAs. The prevalence of RVC in wild boar population reached 12.8%, PTV was detected in 20.2%, PSV in 8.9%, and EV-G in 2.5% of samples. During our study no PRRSV or coronaviruses were detected. Our study provides the first evidence of RVC prevalence in wild boars and indicates that wild boars might contribute to the genetic variability of RVA and also serve as an important reservoir of other enteric viruses.

• MeSH
• antigeny virové genetika   MeSH
• Coronaviridae genetika   izolace a purifikace   MeSH
• feces virologie   MeSH
• fylogeneze MeSH
• genotyp MeSH
• infekce viry z čeledi Coronaviridae epidemiologie   veterinární   virologie   MeSH
• lidé MeSH
• nemoci prasat epidemiologie   virologie   MeSH
• Picornaviridae genetika   izolace a purifikace   MeSH
• pikornavirové infekce epidemiologie   veterinární   virologie   MeSH
• prasata MeSH
• rotavirové infekce epidemiologie   veterinární   virologie   MeSH
• Rotavirus genetika   izolace a purifikace   MeSH
• Sus scrofa MeSH
• virové plášťové proteiny genetika   MeSH
• zdroje nemoci MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH

Porcine reproductive and respiratory syndrome virus (PRRSV) can predispose pigs to secondary respiratory infection with bacteria such as Haemophilus parasuis. Animals infected with both pathogens develop more severe clinical disease. The immune response of porcine alveolar macrophages (PAMs) to simultaneous infection with PRRSV and H. parasuis was analysed in vitro, describing cytokine production, expression of cell surface molecules, and production of reactive oxygen species (ROS). Concurrent infection with PRRSV and H. parasuis increased gene expression of pro-inflammatory cytokines (TNF-α, IL-1β, IL-8) in PAMs in comparison with PAMs infected with PRRSV or H. parasuis alone. An additive effect of dual infection on IL-1β production was confirmed at the protein level. PAMs infected with PRRSV showed increased production of ROS compared to controls. Conversely, simultaneous infection of PAMs with PRRSV and H. parasuis decreased production of ROS, indicating the presence of an H. parasuis defence mechanism against respiratory burst. Concurrent infection of PAMs with PRRSV and H. parasuis was shown to elicit a pro-inflammatory immune response represented by significant IL-1β production. Severe multifactorial respiratory disease in natural conditions caused by both pathogens could be the consequence of pro-inflammatory mediated immunopathology.

• MeSH
• alveolární makrofágy imunologie   mikrobiologie   virologie   MeSH
• biologické markery analýza   MeSH
• cytokiny genetika   imunologie   MeSH
• Haemophilus parasuis imunologie   MeSH
• hemofilové infekce komplikace   imunologie   veterinární   virologie   MeSH
• koinfekce veterinární   MeSH
• kultivované buňky MeSH
• messenger RNA metabolismus   MeSH
• nemoci prasat imunologie   mikrobiologie   virologie   MeSH
• prasata MeSH
• regulace exprese virových genů MeSH
• regulace genové exprese u bakterií MeSH
• reprodukční a respirační syndrom prasat imunologie   virologie   MeSH
• virus reprodukčního a respiračního syndromu prasat imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Rotavirus C (RVC) is a cause of gastroenteritis in swine and has a worldwide distribution. A total of 448 intestinal or faecal samples from pigs of all ages were tested for viruses causing gastroenteritis. RVC was detected in 118 samples (26.3%). To gain information on virus diversity, the complete coding nucleotide sequences of the VP7, VP4, VP6, NSP2, NSP4, and NSP5 genes of seven RVC strains were determined. Phylogenetic analysis of VP7 nucleotide sequence divided studied Czech strains into six G genotypes (G1, G3, G5-G7, and a newly described G10 genotype) based on an 85% identity cutoff value at the nucleotide level. Analysis of the VP4 gene revealed low nucleotide sequence identities between two Czech strains and other porcine (72.2-75.3%), bovine (74.1-74.6%), and human (69.1-69.3%) RVC strains. Thus, we propose that those two Czech porcine strains comprise a new RVC VP4 genotype, P8. Analysis of the VP6 gene showed 79.9-86.8% similarity at the nucleotide level between the Czech strains and other porcine RVC strains. According to the 87% identity cutoff value, we propose the existence of three new RVC VP6 genotypes, I8-I10. Analysis of the NSP4 gene divided porcine RVC strains into two clusters (the E1 genotype and the new E4 genotype, based on an 85% nucleotide sequence identity cutoff value). Our results indicate a degree of high genetic heterogeneity, not only in the variable VP7 and VP4 genes encoding the outer capsid proteins, but also in more-conserved genes encoding the inner capsid protein VP6 and the non-structural proteins NSP2, NSP4, and NSP5. This emphasizes the need for a whole-genome-sequence-based classification system.

• MeSH
• fylogeneze * MeSH
• genetická variace * MeSH
• lidé MeSH
• molekulární sekvence - údaje MeSH
• nemoci prasat virologie   MeSH
• nemoci skotu virologie   MeSH
• prasata MeSH
• rotavirové infekce veterinární   virologie   MeSH
• Rotavirus chemie   klasifikace   genetika   izolace a purifikace   MeSH
• sekvence aminokyselin MeSH
• sekvence nukleotidů MeSH
• sekvenční analýza DNA MeSH
• sekvenční homologie aminokyselin MeSH
• skot MeSH
• virové proteiny chemie   genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

Group C rotavirus (RVC) has been described to be a causative agent of gastroenteritis in humans and animals. In the current study, the presence of porcine RVC was confirmed in 25.6 % of 293 porcine faecal samples collected from seven Czech farms. A significantly larger (p < 0.05) number of RVC-positive samples was detected in groups of finisher pigs and post-weaning piglets (4-12 weeks of age). Phylogenetic analysis of nine RVC-positive Czech strains and their comparison with available sequence data for the gene encoding RVC group antigen VP6 revealed two separate lineages within porcine cluster I1.

• MeSH
• antigeny virové genetika   MeSH
• feces virologie   MeSH
• fylogeneze MeSH
• genetická variace MeSH
• lidé MeSH
• nemoci prasat epidemiologie   virologie   MeSH
• prasata MeSH
• prevalence MeSH
• rotavirové infekce epidemiologie   veterinární   virologie   MeSH
• Rotavirus klasifikace   MeSH
• shluková analýza MeSH
• virové plášťové proteiny genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH