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1.
Microbiol Spectr ; 11(6): e0264323, 2023 Dec 12.
Artículo en Inglés | MEDLINE | ID: mdl-37830808

RESUMEN

IMPORTANCE: Enterovirus G is a species of positive-sense single-stranded RNA viruses associated with several mammalian diseases. The porcine enterovirus strains isolated here were chimeric viruses with the PLCP gene of porcine torovirus, which grouped together with global EV-G1 strains. The isolated EV-G strain could infect various cell types from different species, suggesting its potential cross-species infection risk. Animal experiment showed the pathogenic ability of the isolated EV-G to piglets. Additionally, the EV-Gs were widely distributed in the swine herds. Our findings suggest that EV-G may have evolved a novel mechanism for broad tropism, which has important implications for disease control and prevention.


Asunto(s)
Infecciones por Enterovirus , Enterovirus , Enterovirus Porcinos , Enfermedades de los Porcinos , Porcinos , Animales , Enterovirus Porcinos/genética , Infecciones por Enterovirus/veterinaria , Virulencia , Filogenia , Enterovirus/genética , Mamíferos
2.
J Vet Med Sci ; 85(2): 252-265, 2023 Feb 21.
Artículo en Inglés | MEDLINE | ID: mdl-36543238

RESUMEN

Type 1 recombinant enterovirus G (EV-G), which carries the papain-like cysteine protease (PLCP) gene of torovirus between its 2C/3A regions, and type 2 recombinant EV-G, which carries the torovirus PLCP gene with its flanking regions having non-EV-G sequences in place of the viral structural genes, have been detected in pig farms in several countries. In a previous study, we collected 222 fecal samples from 77 pig farms from 2104 to 2016 and detected one type 2 recombinant EV-G genome by metagenomics sequencing. In this study, we reanalyzed the metagenomic data and detected 11 type 2 recombinant EV-G genomes. In addition, we discovered new type 2 recombinant EV-G genomes of the two strains from two pig farms samples in 2018 and 2019. Thus, we identified the genomes of 13 novel type 2 recombinant EV-Gs isolated from several pig farms in Japan. Type 2 recombinant EV-G has previously been detected only in neonatal piglets. The present findings suggest that type 2 recombinant EV-G replicates in weaning piglets and sows. The detection of type 1 recombinant EV-Gs and type 2 recombinant EV-Gs at 3-year and 2-year intervals, respectively, from the same pig farm suggests that the viruses were persistently infecting or circulating in these farms.


Asunto(s)
Infecciones por Enterovirus , Enterovirus Porcinos , Enfermedades de los Porcinos , Porcinos , Animales , Femenino , Enterovirus Porcinos/genética , Granjas , Infecciones por Enterovirus/veterinaria , Japón , Recombinación Genética , Genoma Viral , Filogenia
3.
Arch Virol ; 165(12): 2909-2914, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-32951133

RESUMEN

Two and three genotypes of enterovirus G (EV-G) carrying a papain-like cysteine protease (PL-CP) sequence were detected on two pig farms and classified into genotypes G1 and G10, and G1, G8, and G17, respectively, based on VP1 sequences. A G10 EV-G virus bearing a PL-CP sequence was detected for the first time. Phylogenetic analysis of the P2 and P3 regions grouped the viruses by farm with high sequence similarity. Furthermore, clear recombination break points were detected in the 2A region, suggesting that PL-CP EV-G-containing strains gained sequence diversity through recombination events among the multiple circulating EV-G genotypes on the farms.


Asunto(s)
Proteasas de Cisteína/genética , Infecciones por Enterovirus/veterinaria , Enterovirus Porcinos/genética , Genoma Viral , Recombinación Genética , Animales , Proteínas de la Cápside/genética , Infecciones por Enterovirus/epidemiología , Infecciones por Enterovirus/virología , Enterovirus Porcinos/enzimología , Heces/virología , Variación Genética , Genotipo , Japón , Filogenia , Análisis de Secuencia de ADN , Sus scrofa , Proteínas Virales/genética
4.
Pathog Dis ; 78(5)2020 07 01.
Artículo en Inglés | MEDLINE | ID: mdl-32691821

RESUMEN

Porcine enterovirus G (EV-G) and teschovirus (PTV) generally cause asymptomatic infections. Although both viruses have been reported from various countries, they are rarely detected from India. To detect these viruses in Western India, fecal samples (n = 26) of diarrheic piglets aged below three months from private pig farms near Pune (Maharashtra) were collected. The samples were screened by reverse transcription-polymerase chain reaction using conserved enterovirus specific primers from 5' untranslated region. For genetic characterization of detected EV-G strain, nearly complete genome, and for PTV, partial VP1 gene were sequenced. EV-G strain showed the highest identity in a VP1 gene at nucleotide (78.61%) and amino acid (88.65%) level with EV-G15, prototype strain. However, its complete genome was homologous with the nucleotide (78.38% identity) and amino acid (91.24% identity) level to Ishi-Ka2 strain (LC316832), unassigned EV-G genotype detected from Japan. The nearly complete genome of EV-G15 consisted of 7398 nucleotides excluding the poly(A) tail and has an open reading frame that encodes a 2170 amino acid polyprotein. Genetic analysis of the partial VP1 gene of teschovirus identified porcine teschovirus 4 (PTV-4) and putative PTV-17 genotype. To the best of our knowledge, this is the first report on nearly full genome characterization of EV-G15, and detection of PTV-4 and putative PTV-17 genotypes from India. Further, detection and characterization of porcine enteroviruses are needed for a comprehensive understanding of their genetic diversity and their association with symptomatic infections from other geographical regions of India.


Asunto(s)
Enterovirus Porcinos/clasificación , Enterovirus Porcinos/genética , Teschovirus/clasificación , Teschovirus/genética , Regiones no Traducidas 5' , Animales , Infecciones Asintomáticas/epidemiología , ADN Viral , Infecciones por Enterovirus/veterinaria , Infecciones por Enterovirus/virología , Enterovirus Porcinos/aislamiento & purificación , Heces/virología , Variación Genética , Genotipo , India/epidemiología , Tipificación Molecular , Sistemas de Lectura Abierta , Filogenia , Infecciones por Picornaviridae/veterinaria , Infecciones por Picornaviridae/virología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Porcinos/virología , Enfermedades de los Porcinos/virología , Teschovirus/aislamiento & purificación , Secuenciación Completa del Genoma
5.
Virol J ; 17(1): 88, 2020 07 01.
Artículo en Inglés | MEDLINE | ID: mdl-32611446

RESUMEN

BACKGROUND: Bovine viral diarrhea virus (BVDV) is a cause of substantial economic loss to the cattle industry worldwide, and there are currently no effective treatment or preventive measures. Bovine enterovirus (BEV) has a broad host range with low virulence and is a good candidate as a viral vaccine vector. In this study, we explored new insertion sites for the expression of exogenous genes in BEV, and developed a recombinant infectious cDNA clone for BEV BJ101 strain expressing BVDV E0 protein. METHODS: A recognition site for the viral proteinase 3Cpro was inserted in the GpBSK-BEV plasmid at the 2C/3A junction by overlapping PCR. Subsequently, the optimized full-length BVDV E0 gene was inserted to obtain the recombinant infectious plasmid GpBSK-BEV-E0. The rescued recombinant virus was obtained by transfection with linearized plasmid. Expression of BVDV E0 in the recombinant virus was confirmed by PCR, western blotting, and immunofluorescence analysis, and the genetic stability was tested in MDBK cells over 10 passages. We further tested the ability of the recombinant virus to induce an antibody response in mice infected with BVDV and immunized them with the recombinant virus and parental strain. RESULTS: The rescued recombinant virus rBEV-E0 was identified and confirmed by western blot and indirect immunofluorescence. The sequencing results showed that the recombinant virus remained stable for 10 passages without genetic changes. There was also no significant difference in growth dynamics and plaque morphology between the recombinant virus and parental virus. Mice infected with both recombinant and parental viruses produced antibodies against BEV VP1, while the recombinant virus also induced antibodies against BVDV E0. CONCLUSION: A new insertion site in the BEV vector can be used for the prevention and control of both BEV and BVDV, providing a useful tool for future research on the development of viral vector vaccines.


Asunto(s)
Anticuerpos Antivirales/sangre , Infecciones por Enterovirus/veterinaria , Enterovirus Bovino/genética , Proteínas del Envoltorio Viral/inmunología , Vacunas Virales/inmunología , Animales , Bovinos , Línea Celular , Virus de la Diarrea Viral Bovina/genética , Virus de la Diarrea Viral Bovina/inmunología , Infecciones por Enterovirus/prevención & control , Femenino , Ratones , Ratones Endogámicos BALB C , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/inmunología , Proteínas del Envoltorio Viral/genética , Vacunas Virales/administración & dosificación , Vacunas Virales/genética
6.
Viruses ; 11(10)2019 10 22.
Artículo en Inglés | MEDLINE | ID: mdl-31652508

RESUMEN

Cynomolgus macaques are common across South East Asian countries including Thailand. The National Primate Research Center of Thailand, Chulalongkorn University (NPRCT-CU) captures wild-borne cynomolgus macaque for research use. Limited information is available on the enteric viruses and possible zoonotic infections into or from cynomolgus macaques. We characterized and compare the fecal virome of two populations; healthy wild-originated captive cynomolgus macaques (n = 43) reared in NPRCT-CU and healthy wild cynomolgus macaques (n = 35). Over 90% of recognized viral sequence reads amplified from feces were from bacterial viruses. Viruses from seven families of mammalian viruses were also detected (Parvoviridae, Anelloviridae, Picornaviridae, Adenoviridae, Papillomaviridae, Herpesviridae, and Caliciviridae). The genomes of a member of a new picornavirus genus we named Mafapivirus, a primate chapparvovirus, and a circular Rep-encoding single-strand (CRESS) DNA virus were also characterized. Higher abundance of CRESS DNA viruses of unknown tropism and invertebrate-tropic ambidensovirus were detected in wild versus captive macaques likely reflecting dietary differences. Short term rearing in captivity did not have a pronounced effect on the diversity of mammalian viruses of wild cynomolgus macaques. This study is the first report of the fecal virome of cynomolgus macaques, non-human primates frequently used in biomedical research and vaccination studies.


Asunto(s)
Animales Salvajes/virología , Animales de Zoológico/virología , Infecciones por Enterovirus/veterinaria , Enterovirus/clasificación , Variación Genética , Macaca fascicularis/virología , Animales , Heces/virología , Femenino , Genoma Viral , Secuenciación de Nucleótidos de Alto Rendimiento , Masculino , Metagenómica , Filogenia , Tailandia
7.
Infect Genet Evol ; 75: 103975, 2019 11.
Artículo en Inglés | MEDLINE | ID: mdl-31344488

RESUMEN

Enterovirus G (EV-G) belongs to the family of Picornaviridae. Two types of recombinant porcine EV-Gs carrying papain-like cysteine protease (PLCP) gene of porcine torovirus, a virus in Coronaviridae, are reported. Type 1 recombinant EV-Gs are detected in pig feces in Japan, USA, and Belgium and carry the PLPC gene at the junction site of 2C/3A genes, while PLPC gene replaces the viral structural genes in type 2 recombinant EV-G detected in pig feces in a Chinese farm. We identified a novel type 2 recombinant EV-G carrying the PLCP gene with flanking sequences in place of the viral structural genes in pig feces in Japan. The ~0.3 kb-long upstream flanking sequence had no sequence homology with any proteins deposited in GenBank, while the downstream ~0.9 kb-long flanking sequence included a domain having high amino acid sequence homology with a baculoviral inhibitor of apoptosis repeat superfamily. The pig feces, where the novel type 2 recombinant EV-G was detected, also carried type 1 recombinant EV-G. The amount of type 1 and type 2 recombinant EV-G genomes was almost same in the pig feces. Although the phylogenetic analysis suggested that these two recombinant EV-Gs have independently evolved, type 1 recombinant EV-G might have served as a helper virus by providing viral structural proteins for dissemination of the type 2 recombinant EV-G.


Asunto(s)
Proteínas Reguladoras de la Apoptosis/metabolismo , Proteasas de Cisteína/genética , Infecciones por Enterovirus/veterinaria , Enterovirus Porcinos/genética , Proteínas Estructurales Virales/genética , Animales , Proteínas Reguladoras de la Apoptosis/genética , Proteasas de Cisteína/metabolismo , Infecciones por Enterovirus/epidemiología , Infecciones por Enterovirus/virología , Heces/virología , Regulación Enzimológica de la Expresión Génica , Regulación Viral de la Expresión Génica , Filogenia , Porcinos , Enfermedades de los Porcinos/epidemiología , Enfermedades de los Porcinos/virología
8.
PLoS One ; 14(3): e0213295, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-30830947

RESUMEN

Viral respiratory and intestinal infections are the most common causes of canine viral illness. Infection with multiple pathogens occurs in many cases. Rapid diagnosis of these multiple infections is important for providing timely and effective treatment. To improve diagnosis, in this study, two new multiplex polymerase chain reactions (mPCRs) were developed for simultaneous detection of canine respiratory viruses (CRV) and canine enteric viruses (CEV) using two separate primer mixes. The viruses included canine adenovirus type 2 (CAV-2), canine distemper virus (CDV), canine influenza virus (CIV), canine parainfluenza virus (CPIV), canine circovirus (CanineCV), canine coronavirus (CCoV) and canine parvovirus (CPV). The sensitivity of the mPCR results showed that the detection limit of both mPCR methods was 1×104 viral copies. Twenty nasal swabs (NS) and 20 anal swabs (AS) collected from dogs with symptoms of respiratory disease or enteric disease were evaluated using the novel mPCR methods as a clinical test. The mPCR protocols, when applied to these respiratory specimens and intestinal samples, could detect 7 viruses simultaneously, allowing rapid investigation of CRV (CAV-2, CDV, CIV and CPIV) and CEV (CAV-2, CanineCV, CCoV and CPV) status and prompt evaluation of coinfection. Our study provides an effective and accurate tool for rapid differential diagnosis and epidemiological surveillance in dogs.


Asunto(s)
Infecciones por Coronavirus/veterinaria , Coronavirus Canino/aislamiento & purificación , Enfermedades de los Perros/epidemiología , Infecciones por Enterovirus/veterinaria , Enterovirus/aislamiento & purificación , Reacción en Cadena de la Polimerasa Multiplex/métodos , Trastornos Respiratorios/veterinaria , Animales , Infecciones por Coronavirus/virología , Coronavirus Canino/clasificación , Coronavirus Canino/genética , Enfermedades de los Perros/virología , Perros , Enterovirus/clasificación , Enterovirus/genética , Infecciones por Enterovirus/virología , Trastornos Respiratorios/virología
9.
Transbound Emerg Dis ; 66(2): 1023-1028, 2019 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-30431236

RESUMEN

Enterovirus species G (EV-G) comprises a highly diversity of 20 genotypes that is prevalent in pig populations, with or without diarrhoea. In the present study, a novel EV-G strain (KOR/KNU-1811/2018) that resulted from cross-order recombination was discovered in diagnostic faecal samples from neonatal pigs with diarrhoea that were negative for swine enteric coronaviruses and rotavirus. The recombinant EV-G genome possessed an exogenous 594-nucleotide (198-amino acid) sequence, flanked by two viral 3Cpro cleavage sites at the 5' and 3' ends in its 2C/3A junction region. This insertion encoded a predicted protease similar to the porcine torovirus papain-like cysteine protease (PLCP), which was recently found in the EV-G1, -G2, and -G17 genomes. The complete KNU-1811 genome shared 73.7% nucleotide identity with a prototype EV-G1 strain, but had 83.9%-86.7% sequence homology with the global EV-G1-PLCP strains. Genetic and phylogenetic analyses demonstrated that the Korean recombinant EV-G's own VP1 and inserted foreign PLCP genes are most closely related independently to contemporary chimeric G1-PLCP and G17-PLCP strains respectively. These results implied that the torovirus-derived PLCP gene might have undergone continuous nucleotide mutations in the respective EV-G genome following its independent acquisition through naturally occurring recombination. Our results advance the understanding of the genetic evolution of EV-G driven by infrequent viral recombination events, by which EV-G populations laterally gain an exotic gene encoding a virulence factor from heterogeneous virus families, thereby causing clinical disease in swine.


Asunto(s)
Proteasas de Cisteína/genética , Diarrea/veterinaria , Infecciones por Enterovirus/veterinaria , Enterovirus Porcinos/genética , Virus Reordenados/genética , Recombinación Genética , Torovirus/genética , Secuencia de Aminoácidos , Animales , Diarrea/epidemiología , Diarrea/virología , Infecciones por Enterovirus/epidemiología , Infecciones por Enterovirus/virología , Enterovirus Porcinos/aislamiento & purificación , Evolución Molecular , Heces/virología , Genoma Viral , Genotipo , Filogenia , República de Corea/epidemiología , Homología de Secuencia , Porcinos , Enfermedades de los Porcinos
10.
Braz. j. microbiol ; 49(4): 790-794, Oct.-Dec. 2018. tab, graf
Artículo en Inglés | LILACS | ID: biblio-974287

RESUMEN

ABSTRACT Although the use of vaccines has controlled enteric diseases in dogs in many developed countries, vaccine coverage is still under optimal situation in Brazil. There is a large population of nonimmunized dogs and few studies about the identification of the viruses associated with diarrhea. To address this situation, stool samples from 325 dogs were analyzed by polymerase chain reaction for the detection of common enteric viruses such as Canine adenovirus (CAdV), Canine coronavirus (CCoV), Canine distemper virus (CDV), Canine rotavirus (CRV) and Carnivorous protoparvovirus 1 (canine parvovirus 2; CPV-2). At least one of these species was detected in 56.6% (184/325) of the samples. The viruses detected most frequently in either diarrheic or nondiarrheic dog feces were CPV-2 (54.3% of the positive samples), CDV (45.1%) and CCoV (30.4%), followed by CRV (8.2%) and CAdV (4.9%). Only one agent was detected in the majority of the positive samples (63%), but co-infections were present in 37% of the positive samples and mainly included CDV and CPV-2. The data presented herein can improve the clinical knowledge in regions with low vaccine coverage and highlight the need to improve the methods used to control these infectious diseases in domestic dogs.


Asunto(s)
Animales , Perros , Enterovirus/aislamiento & purificación , Enfermedades de los Perros/virología , Infecciones por Enterovirus/veterinaria , Filogenia , Brasil , Vacunas Virales/administración & dosificación , Vacunas Virales/genética , Vacunas Virales/inmunología , Enterovirus/clasificación , Enterovirus/genética , Enfermedades de los Perros/inmunología , Enfermedades de los Perros/prevención & control , Infecciones por Enterovirus/inmunología , Infecciones por Enterovirus/prevención & control , Infecciones por Enterovirus/virología , Heces/virología
11.
Braz J Microbiol ; 49(4): 790-794, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-29588198

RESUMEN

Although the use of vaccines has controlled enteric diseases in dogs in many developed countries, vaccine coverage is still under optimal situation in Brazil. There is a large population of nonimmunized dogs and few studies about the identification of the viruses associated with diarrhea. To address this situation, stool samples from 325 dogs were analyzed by polymerase chain reaction for the detection of common enteric viruses such as Canine adenovirus (CAdV), Canine coronavirus (CCoV), Canine distemper virus (CDV), Canine rotavirus (CRV) and Carnivorous protoparvovirus 1 (canine parvovirus 2; CPV-2). At least one of these species was detected in 56.6% (184/325) of the samples. The viruses detected most frequently in either diarrheic or nondiarrheic dog feces were CPV-2 (54.3% of the positive samples), CDV (45.1%) and CCoV (30.4%), followed by CRV (8.2%) and CAdV (4.9%). Only one agent was detected in the majority of the positive samples (63%), but co-infections were present in 37% of the positive samples and mainly included CDV and CPV-2. The data presented herein can improve the clinical knowledge in regions with low vaccine coverage and highlight the need to improve the methods used to control these infectious diseases in domestic dogs.


Asunto(s)
Enfermedades de los Perros/virología , Infecciones por Enterovirus/veterinaria , Enterovirus/aislamiento & purificación , Animales , Brasil , Enfermedades de los Perros/inmunología , Enfermedades de los Perros/prevención & control , Perros , Enterovirus/clasificación , Enterovirus/genética , Infecciones por Enterovirus/inmunología , Infecciones por Enterovirus/prevención & control , Infecciones por Enterovirus/virología , Heces/virología , Filogenia , Vacunas Virales/administración & dosificación , Vacunas Virales/genética , Vacunas Virales/inmunología
12.
J Gen Virol ; 98(6): 1305-1310, 2017 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-28590234

RESUMEN

Identification of unknown pathogens in pigs displaying enteric illness is difficult due to the large diversity of bacterial and viral species found within faecal samples. Current methods often require bacterial or viral isolation, or testing only a limited number of known species using quantitative PCR analysis. Herein, faeces from two 25-day-old piglets with diarrhoea from Texas, USA, were analysed by metagenomic next-generation sequencing to rapidly identify possible pathogens. Our analysis included a bioinformatics pipeline of rapid short-read classification and de novo genome assembly which resulted in the identification of a porcine enterovirus G (EV-G), a complete genome with substantial nucleotide differences (>30 %) among current sequences, and a novel non-structural protein similar in sequence to the Torovirus papain-like cysteine protease (PLpro). This discovery led to the identification and circulation of an EV-G with a novel PLpro in the USA that has not been previously reported.


Asunto(s)
Proteasas de Cisteína/genética , Diarrea/veterinaria , Infecciones por Enterovirus/veterinaria , Enterovirus Porcinos/clasificación , Enterovirus Porcinos/enzimología , Heces/virología , Enfermedades de los Porcinos/virología , Animales , Análisis por Conglomerados , Biología Computacional , Diarrea/virología , Infecciones por Enterovirus/virología , Enterovirus Porcinos/genética , Enterovirus Porcinos/aislamiento & purificación , Genoma Viral , Secuenciación de Nucleótidos de Alto Rendimiento , Filogenia , Análisis de Secuencia de ADN , Porcinos , Texas
13.
Intervirology ; 59(2): 69-73, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27694750

RESUMEN

OBJECTIVE: The aim of this study was to investigate the exposure of piglets to enteroviruses-G (EV-G) through the presence of antibodies in their serum. METHODS: Serum samples were obtained from the vena cava of 10 piglets at 9 weeks of age and again 39 days later (day 39). They were tested using an immunoassay based on the EV-G1 VP4 peptide, since VP4 is highly conserved among the four Enterovirus capsid proteins, and by using a seroneutralization assay. RESULTS: For each serum collected on day 39 the optical density was high compared to the value obtained in serum collected earlier (p = 0.002). However, the titers of anti-EV-G1 serum neutralizing activity were not different in paired samples (p > 0.999). The sequence alignment of the EV-G1 VP4 peptide, encompassing 50 amino acids, used in the immunoassay showed 88% homology with EV-G, suggesting that antibodies directed toward other EV-G than EV-G1 may be detected. CONCLUSION: An immunoassay based on EV-G1 VP4 can detect an increased level of EV-G antibodies in piglet serum samples. Further studies are needed to determine whether this immunoassay may be useful for diagnosis and/or epidemiological studies and to monitor EV-G infection in pigs to evaluate strategies aimed to prevent enterovirus infections.


Asunto(s)
Anticuerpos Antivirales/sangre , Infecciones por Enterovirus/veterinaria , Enterovirus/inmunología , Enfermedades de los Porcinos/diagnóstico , Proteínas Estructurales Virales/inmunología , Animales , Anticuerpos Neutralizantes , Infecciones por Enterovirus/diagnóstico , Infecciones por Enterovirus/inmunología , Infecciones por Enterovirus/virología , Pruebas de Neutralización , Péptidos/inmunología , Alineación de Secuencia , Sus scrofa , Porcinos , Enfermedades de los Porcinos/inmunología , Enfermedades de los Porcinos/virología
14.
Sci Rep ; 6: 28526, 2016 06 22.
Artículo en Inglés | MEDLINE | ID: mdl-27329349

RESUMEN

Recent studies of Enterovirus (EV) in nonhuman primates (NHPs), which could act as a source of future emerging human viral diseases, have boosted interest in the search for novel EVs. Here, a highly divergent strain of EV, tentatively named SEV-gx, was identified by viral metagenomic analysis from stool samples of rhesus macaques in China. In total, 27 of 280 (9.6%) faecal samples from rhesus macaques were positive for SEV-gx. Its complete genomic sequence is 7,367 nucleotide (nt). Genomic analyses showed that it has a standard genomic organisation for EVs, being more closely related to EV-J strains (approximately 54.0%, 43.0-44.1%, 52.3-55.2%, 61.1-62.7% and 64.0% amino acids identity in polyprotein, P1, P2 and P3 and combined 2C/3CD regions, respectively). It was also shown to have genome characteristics typical of EVs. Phylogenetic analysis of P1, 2C and 3CD aa indicated that SEV-gx can be classified as a distinct cluster in the EVs. All of this evidence demonstrates SEV-gx is a novel species (tentatively named EV-K) in the EV genus, which contributes to our understanding of the genetic diversity and evolution of EVs. Further studies are needed to investigate the potential pathogenicity of SEV-gx in NHPs and humans.


Asunto(s)
Infecciones por Enterovirus/veterinaria , Enterovirus/clasificación , Macaca mulatta/virología , Enfermedades de los Monos/virología , Animales , China , Enterovirus/genética , Enterovirus/aislamiento & purificación , Infecciones por Enterovirus/virología , Evolución Molecular , Heces/virología , Variación Genética , Genoma Viral , Humanos , Metagenómica , Conformación de Ácido Nucleico , Filogenia , ARN Viral/química , ARN Viral/genética , Análisis de Secuencia de ARN , Proteínas Estructurales Virales/genética
15.
J Gen Virol ; 97(2): 378-388, 2016 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-26653281

RESUMEN

A recent survey of pigs in Dong Thap province, Vietnam identified a high frequency of enterovirus species G (EV-G) infection (144/198; 72.7%). Amongst these was a plethora of EV-G types (EV-G1, EV-G6 and four new types EV-G8-EV-G11). To better characterize the genetic diversity of EV-G and investigate the possible existence of further circulating types, we performed a larger-scale study on 484 pig and 45 farm-bred boar faecal samples collected in 2012 and 2014, respectively. All samples from the previous and current studies were also screened for kobuviruses. The overall EV infection frequency remained extremely high (395/484; 81.6%), but with comparable detection rates and viral loads between healthy and diarrhoeic pigs; this contrasted with less frequent detection of EV-G in boars (4/45; 8.9%). EV was most frequently detected in pigs ≤ 14 weeks old (∼ 95%) and declined in older pigs. Infections with EV-G1 and EV-G6 were most frequent, whilst less commonly detected types included EV-G3, EV-G4 and EV-G8-EV-G11, and five new types (EV-G12-EV-G16). In contrast, kobuvirus infection frequency was significantly higher in diarrhoeic pigs (40.9 versus 27.6%; P = 0.01). Kobuviruses also showed contrasting epizootiologies and age associations; a higher prevalence was found in boars (42%) compared with domestic pigs (29%), with the highest infection frequency amongst pigs >52 weeks old. Although genetically diverse, all kobuviruses identified belonged to the species Aichivirus C. In summary, this study confirms infection with EV-G was endemic in Vietnamese domestic pigs and exhibits high genetic diversity and extensive inter-type recombination.


Asunto(s)
Infecciones por Enterovirus/veterinaria , Enterovirus/aislamiento & purificación , Heces/virología , Kobuvirus/aislamiento & purificación , Infecciones por Picornaviridae/veterinaria , Enfermedades de los Porcinos/epidemiología , Enfermedades de los Porcinos/virología , Animales , Diarrea/epidemiología , Diarrea/veterinaria , Diarrea/virología , Enterovirus/clasificación , Enterovirus/genética , Infecciones por Enterovirus/epidemiología , Infecciones por Enterovirus/virología , Variación Genética , Kobuvirus/clasificación , Kobuvirus/genética , Tamizaje Masivo , Infecciones por Picornaviridae/epidemiología , Infecciones por Picornaviridae/virología , Prevalencia , Sus scrofa , Porcinos , Vietnam/epidemiología , Carga Viral
16.
Braz J Biol ; 75(2 Suppl): 11-6, 2015 May.
Artículo en Inglés | MEDLINE | ID: mdl-26270208

RESUMEN

The spread of enteric viruses of domestic animals and human beings to wild species can be facilitated by the resistance of these viruses on the environment and their ability to be transmitted by water and contaminated food. The health status of the populations of pampas foxes Lycalopex gymnocercus) and crab-eating foxes (Cerdocyon thous) is largely unknown and the landscapes occupied by these animals in southern Brazil have been threatened by human occupation and expansion of agriculture. In this work, the search of genomes of human and canine adenoviruses in feces from these wild carnivores was used to track the dissemination of domestic animals and human pathogens to the free-living populations in a wildlife reserve located in southern Brazil. This was performed by virus-specific differential real-time polymerase chain reactions (qPCR) on stool specimens, avoiding capture and additional stress to the animals. Genus-specific conventional reverse-transcriptase PCR (RT-PCR) was complementarily performed aiming the detection of enteroviruses (EV) and rotaviruses (RV) on these same samples. HAdV genomes were found on 14 out of the 17 (82.35%) stool samples analysed, whereas CAV was found co-infecting 5 of these samples. RV genomes were detected on 7 of the 17 samples (41.18%) and all samples were negative for EV. The results point to the dispersion of HAdV and RV at a high rate to these species of South American wild carnivores, which can be an effect of growing anthropisation of the habitat of these animals.


Asunto(s)
Infecciones por Adenoviridae/veterinaria , Adenoviridae/aislamiento & purificación , Zorros , Adenoviridae/genética , Infecciones por Adenoviridae/virología , Animales , Brasil , Perros , Enterovirus/aislamiento & purificación , Infecciones por Enterovirus/genética , Infecciones por Enterovirus/veterinaria , Infecciones por Enterovirus/virología , Heces/virología , Humanos , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Rotavirus/aislamiento & purificación , Infecciones por Rotavirus/genética , Infecciones por Rotavirus/veterinaria , Infecciones por Rotavirus/virología , Especificidad de la Especie
17.
Braz. j. biol ; 75(2,supl): 11-16, May 2015. tab
Artículo en Inglés | LILACS | ID: lil-755027

RESUMEN

The spread of enteric viruses of domestic animals and human beings to wild species can be facilitated by the resistance of these viruses on the environment and their ability to be transmitted by water and contaminated food. The health status of the populations of pampas foxes (Lycalopex gymnocercus) and crab-eating foxes (Cerdocyon thous) is largely unknown and the landscapes occupied by these animals in southern Brazil have been threatened by human occupation and expansion of agriculture. In this work, the search of genomes of human and canine adenoviruses in feces from these wild carnivores was used to track the dissemination of domestic animals and human pathogens to the free-living populations in a wildlife reserve located in southern Brazil. This was performed by virus-specific differential real-time polymerase chain reactions (qPCR) on stool specimens, avoiding capture and additional stress to the animals. Genus-specific conventional reverse-transcriptase PCR (RT-PCR) was complementarily performed aiming the detection of enteroviruses (EV) and rotaviruses (RV) on these same samples. HAdV genomes were found on 14 out of the 17 (82.35%) stool samples analysed, whereas CAV was found co-infecting 5 of these samples. RV genomes were detected on 7 of the 17 samples (41.18%) and all samples were negative for EV. The results point to the dispersion of HAdV and RV at a high rate to these species of South American wild carnivores, which can be an effect of growing anthropisation of the habitat of these animals.

.

A disseminação de vírus entéricos de animais domésticos e seres humanos para espécies selvagens pode ser facilitada pela resistência desses vírus no ambiente e sua capacidade de ser transmitida por água e alimentos contaminados. O estado de saúde das populações de Graxains-do-campo (Lycalopex gymnocercus) e Cachorros-do-mato (Cerdocyon thous) é em grande parte desconhecida e as paisagens ocupadas por estes animais no sul do Brasil têm sido ameaçadas pela ocupação humana e a expansão da agricultura. Neste trabalho, utilizou-se a pesquisa de genomas de adenovírus humanos (HAdV ) e caninos (CAV-1 e -2) em amostras fezes desses carnívoros selvagens com vistas a diagnosticar a disseminação de patógenos de animais domésticos e seres humanos às populações de vida livre em uma reserva de vida selvagem, localizado no sul do Brasil. Foram realizadas reações em cadeia da polimerase diferenciais e em tempo real (qPCR) de adenovírus específicos em amostras de fezes, evitando a captura e estresse adicional para os animais. PCRs gênero-específicas convencionais com transcrição reversa prévia (RT-PCR) foram ainda realizadas visando a detecção de enterovírus (EV) e rotavírus (RV) nestas mesmas amostras. Genomas de HAdV foram encontrados em 14 a 17 amostras de fezes (82.35%) analisados, Considerando que o CAV foi encontrado coinfectando 5 destas amostras. Genomas de RV foram detectados em 7 das 17 amostras (41.18%) e todas as amostras foram negativas para EV. Os resultados apontam para a dispersão de HAdV e RV em uma taxa elevada para estas espécies de carnívoros selvagens sul-americanas, que podem ser um efeito da crescente antropização do habitat desses animais.

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Asunto(s)
Animales , Perros , Humanos , Infecciones por Adenoviridae/veterinaria , Adenoviridae/aislamiento & purificación , Zorros , Infecciones por Adenoviridae/virología , Adenoviridae/genética , Brasil , Infecciones por Enterovirus/genética , Infecciones por Enterovirus/veterinaria , Infecciones por Enterovirus/virología , Enterovirus/aislamiento & purificación , Heces/virología , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Infecciones por Rotavirus/genética , Infecciones por Rotavirus/veterinaria , Infecciones por Rotavirus/virología , Rotavirus/aislamiento & purificación , Especificidad de la Especie
18.
PLoS One ; 9(5): e97730, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24830424

RESUMEN

In this study, a virus strain designated as HY12 was isolated from cattle with a disease of high morbidity and mortality in Jilin province. Biological and physiochemical properties showed that HY12 isolates is cytopathic with an extremely high infectivity. HY12 is resistant to treatment of organic solvent and acid, and unstable at 60°C for 1 h. Electron microscopy observation revealed the virus is an approximately 22-28 nm in diameter. The complete genome sequence of HY12 consists of 7416 nucleotides, with a typical picornavirus genome organization including a 5'-untranslated region (UTR), a large single ORF encoding a polyprotein of 2176 amino acids, and a 3'-UTR. Phylogenetic analysis clustered HY12 isolates to a new serotype/genotype within the clade of enterovirus E (formerly BEV-A). Alignment analysis revealed a unique insertion of 2 amino acid residues (NF) at the C-terminal of VP1 protein between aa 825 and 826, and several rare mutations in VP1 and VP4 of HY12 isolates in relation to known bovine enterovirus (BEV) strains. This is the first report of an enterovirus E in China, which is potentially associated with an outbreak in cattle with severe respiratory and enteric diseases.


Asunto(s)
Enfermedades de los Bovinos/virología , Enteritis/veterinaria , Infecciones por Enterovirus/veterinaria , Enterovirus Bovino/genética , Infecciones del Sistema Respiratorio/veterinaria , Secuencia de Aminoácidos , Animales , Proteínas de la Cápside/química , Bovinos , Línea Celular , Enteritis/virología , Infecciones por Enterovirus/virología , Enterovirus Bovino/clasificación , Enterovirus Bovino/aislamiento & purificación , Genotipo , Datos de Secuencia Molecular , Filogenia , Infecciones del Sistema Respiratorio/virología , Análisis de Secuencia de ADN
19.
J Virol ; 86(12): 7008-9, 2012 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-22628395

RESUMEN

The porcine enteroviruses (PEVs) belong to the family Picornaviridae. We report a complete genome sequence of a novel PEV strain that is widely prevalent in pigs at least in central and eastern China. The complete genome consists of 7,390 nucleotides, excluding the 3' poly(A) tail, and has an open reading frame that maps between nucleotide positions 812 and 7318 and encodes a 2,168-amino-acid polyprotein. Phylogenetic analysis based on the 3CD and VP1 regions reveals that this PEV strain belongs to a species of PEV9 but may represent a novel sero-/genotype in CPE group III. We also report the major findings from bootscan analysis based on the whole genomes of PEVs in the present study and those available in GenBank.


Asunto(s)
Infecciones por Enterovirus/veterinaria , Enterovirus Porcinos/genética , Genoma Viral , Enfermedades de los Porcinos/virología , Animales , Secuencia de Bases , China , Infecciones por Enterovirus/virología , Enterovirus Porcinos/clasificación , Enterovirus Porcinos/aislamiento & purificación , Datos de Secuencia Molecular , Filogenia , Porcinos
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