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1.
Arch Virol ; 165(1): 137-143, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31745718

RESUMEN

Canine circovirus (canine CV) is an etiological agent associated with diarrhea, hemorrhagic gastroenteritis and vasculitis. Although canine CV has been identified and characterized in southern China in recent years, its epidemiology in other regions of China and its precise molecular characteristics have not been examined. In this study, we examined 141 fecal specimens collected from domestic dogs with or without diarrhea in Heilongjiang province, Northeastern China, during 2014 to 2016. A total of 18 out of 141 samples were found to be positive for canine CV by real-time quantitative PCR. In the diarrhea samples, canine CV was detected in coinfections with canine parvovirus 2. More importantly, two different canine CV strains were detected in one sample. Five canine CV genomes were successfully amplified. Sequence analysis showed that there were two unique amino acid changes in the Rep protein (N39S in the K1 strain, and T71A in the XF16 strain). Phylogenetic analysis indicated that canine CV could be divided into four genotypes, and specific nucleotide mutations could be used for confirming the four genotypes. Moreover, recombination analysis revealed that a total of eight recombination events were found in five genomic sequences. Molecular evolution analysis showed that the canine CV has been under purifying selection. This study provides evidence that at least three genotypes of canine CV are co-circulating in China. Continuous epidemiological surveillance is therefore necessary to understand their importance for the evolution of canine CV.


Asunto(s)
Circovirus/clasificación , Diarrea/veterinaria , Enfermedades de los Perros/virología , Mutación , Parvovirus Canino/aislamiento & purificación , Animales , China , Circovirus/genética , Diarrea/virología , Perros , Evolución Molecular , Heces/virología , Parvovirus Canino/genética , Filogenia , Proteínas Virales/genética , Secuenciación Completa del Genoma
2.
Food Microbiol ; 85: 103307, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31500711

RESUMEN

Cold atmospheric-gaseous plasma (CAP) is an emerging non-thermal technology for decontamination of foodborne bacterial and viral pathogens. We obtained a >5 log10 reduction in the titer (TCID50) of feline calicivirus (FCV) on stainless steel discs and Romaine lettuce leaves after 3 min wet exposure to air plasma generated by a two-dimensional array of integrated coaxial-microhollow dielectric barrier discharge (2D-AICM-DBD). However, when human norovirus (HuNoV GII.4) was treated for 5 min under the same conditions, ~2.6 log10 (>99.5%) reduction in genome copy number was observed as measured by ethidium monoazide-coupled RT-qPCR (EMA-RT-qPCR). To assess this discrepancy, we studied CAP's effect on FCV by the cell culture method and by the EMA-coupled RT-qPCR method. It was found that the molecular titration method (EMA-RT-qPCR) underestimates the level of virus reduction by CAP. Additionally, the fecal matter present in HuNoV samples partially suppressed virucidal activity of CAP. Assuming that the lower virus reduction measured by EMA-RT-qPCR method compared to cell culture method for FCV is the same as for HuNoV, we can conclude that FCV may be used as a surrogate for HuNoV to assess the virucidal effect of CAP. CAP is able to inactivate 3.5 Log10 units of HuNoV at low titers after 2 min of exposure.


Asunto(s)
Heces/virología , Norovirus/efectos de los fármacos , Gases em Plasma/farmacología , Inactivación de Virus/efectos de los fármacos , Azidas , Calicivirus Felino/efectos de los fármacos , Calicivirus Felino/genética , Desinfección/métodos , Humanos , Lechuga/virología , Norovirus/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Acero Inoxidable
3.
Sci Total Environ ; 700: 134446, 2020 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-31648121

RESUMEN

Bacteriophage may play an important role in antimicrobial resistance genes (ARGs) transmission. However, the contribution of bacteriophage to the spread of ARGs in environment, especially in poultry farm environment, is rarely known. In this study, the prevalence of ARGs in bacteriophage DNA was investigated in chicken feces from 30 different poultry farms in China. Then the abundance of the aac(6')-Ib-cr, blaCTX-M, ermB, floR, mcr-1, sul1, tetM and intI1 genes was determined by qPCR in bacteriophage and compared with certain representative plasmid DNA samples. The results showed that 12 ARGs (aac(6')-Ib-cr, aph(3')-IIIa, blaCTX-M, ermB, ermF, floR, mcr-1, qnrS, sul1, sul2, vanA, tetM genes) and class 1 integron gene intI1 were detected in bacteriophage DNA fraction. The sul1, tetM and aac(6')-Ib-cr genes were most prevalent with high detection rates of 77%, 61% and 55%, respectively. To our best knowledge, this study firstly reported the presence of the mcr-1 gene in bacteriophage DNA derived from farms environments. We found that the gene copy (GC) numbers of the aac(6')-Ib-cr, ermB and sul1 genes were as high as 5.47, 5.22 and 5.54 log10 GC/g, respectively. Both the prevalence and abundance of ARGs in broiler fecal wastes were also generally higher than in laying hens. In addition, although the GC numbers of the aac(6')-Ib-cr, floR and tetM genes in plasmid DNA was higher than that in phage DNA fraction by 4.68, 3.59 and 3.9 orders of magnitude, respectively, the absolute abundances of the blaCTX-M and mcr-1 genes in phage DNA were close to or even higher than that in plasmid DNA at farm SIL2, SIL4 and SIB1. As potential vessels for ARGs, bacteriophage could not be ignored due to their unique extracellular persistence in environments. Overall, this is the first comprehensive survey about bacteriophage carried ARGs from farms in different regions in China.


Asunto(s)
Farmacorresistencia Bacteriana/genética , Heces/virología , Genes Bacterianos , Animales , Bacteriófagos/genética , Pollos , China , Granjas , Integrones , Plásmidos
4.
Arch Virol ; 165(2): 471-477, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31863265

RESUMEN

We sequenced the complete genome of a porcine torovirus (PToV) strain from Japan for the first time. Whole-genome analysis revealed that this strain (Iba/2018) has a mosaic sequence composed of at least three genome backgrounds, related to US, Chinese and German PToV strains. Clear recombination breakpoints were detected in the M and HE coding regions. A similarity plot and structural analysis demonstrated that the HE coding region exhibits the highest diversity, and the most sequence variation was found in the lectin domain. PToVs were divided into two lineages in the HE region, whereas clear lineages were not found in other regions.


Asunto(s)
Heces/virología , Genoma Viral , Infecciones por Torovirus/veterinaria , Torovirus/genética , Torovirus/aislamiento & purificación , Secuenciación Completa del Genoma , Animales , Biología Computacional , Evolución Molecular , Humanos , Japón , Recombinación Genética , Porcinos , Infecciones por Torovirus/virología
5.
Arch Virol ; 165(2): 463-470, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31863266

RESUMEN

The aim of this work was the genetic typing of RVA isolates originating from pigs and human patients in Slovakia. Seventy-eight rectal swabs from domestic pigs and 30 stool samples from humans were collected. The whole VP7 (G genotypes), VP6 (I genotypes) and partial VP4 (P genotypes) ORFs were amplified by RT-PCR. Genetic variability was higher amongst porcine sequences, where four G genotypes (G3, G4, G5, G11), two P genotypes (P[6], P[13]) and one I5 genotype were detected. Human RVA strains were represented by two G genotypes (G1, G3), two I genotypes (I1, I2), and one P genotype (P[8]). Genetic analysis did not show a relationship between Slovakian porcine and human RVA strains, but phylogenetic grouping of some Slovakian porcine sequences with Hungarian human sequences in both G and P genotypes was observed.


Asunto(s)
Variación Genética , Infecciones por Rotavirus/veterinaria , Infecciones por Rotavirus/virología , Rotavirus/clasificación , Rotavirus/genética , Enfermedades de los Porcinos/virología , Animales , Antígenos Virales/genética , Proteínas de la Cápside/genética , Heces/virología , Genotipo , Humanos , Recto/virología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Rotavirus/aislamiento & purificación , Análisis de Secuencia de ADN , Eslovaquia , Sus scrofa , Porcinos
6.
Arch Virol ; 165(2): 495-504, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31845155

RESUMEN

The discovery of novel viruses in wild animals allows the prediction of their potential threat to the health of humans and other animals. We report a highly divergent picornavirus (tentatively named "mobovirus A"), identified in a fecal sample from Macaca mulatta in Yunnan province, China, using viral metagenomic analysis, with viral loads of 2 × 107 copies/g. The complete genomic sequence of mobovirus A is 8,325 nucleotides in length. Phylogenetic analysis showed that it clustered with Guangxi changeable lizard picornavirus 1 and Guangxi Chinese leopard gecko picornavirus, with less than 38%, 40%, and 40% amino acid identity in the P1, P2, and P3 protein, respectively. The viruses in this cluster were most closely related to members of the genera Harkavirus, Tremovirus and Hepatovirus. Genomic analysis revealed that mobovirus A has the typical genomic organization and motifs of a picornavirus. Additionally, its codon usage bias complements that of M. mulatta, suggesting that this feature is not restricted only to hepatoviruses. Thus, according to the guidelines of the Picornaviridae Study Group of the International Committee on Taxonomy of Viruses, mobovirus A should be considered a member of a new genus (tentatively named for Monkey-borne virus, "Mobovirus") in the family Picornaviridae. These data will facilitate the understanding of the genetic diversity and evolution of picornaviruses. Further studies are needed to understand the epidemiology and potential pathogenicity of the virus in M. mulatta.


Asunto(s)
Macaca mulatta/virología , Picornaviridae/genética , Picornaviridae/aislamiento & purificación , Regiones no Traducidas 5'/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , China , Heces/virología , Genoma Viral/genética , Conformación de Ácido Nucleico , Filogenia , ARN Viral/genética , Proteínas Virales/genética
7.
BMC Infect Dis ; 19(1): 1021, 2019 Dec 02.
Artículo en Inglés | MEDLINE | ID: mdl-31791261

RESUMEN

BACKGROUND: Currently, norovirus (NoV) is associated with one-fifth of all acute gastroenteritis (AGE) cases worldwide. The NoV GII.17_2014 variant has been associated with gastroenteritis outbreaks in several Asian countries, replacing the previously dominant Sydney 2012 variant. There is limited data about circulation of this new strain in Brazil. This study aimed to describe the phylogenetic and evolutionary characteristics of the GII.17_2014 strains in the Northern region of Brazil. METHODS: NoV was detected by enzyme immunoassay (EIA) in 645 stool samples of AGE cases that were reported in Pará and Amazonas states during 2015-2016. All positive samples were tested for NoV GI and GII by reverse transcription polymerase chain reaction (RT-PCR) and the amplicons were subjected to genome sequencing. The GII.17-positive samples were retested by PCR using different sets of designed primers, which target a highly conserved capsid gene region. Next, the amplicons were sequenced and phylogenetically analyzed using Bayesian inferences. RESULTS: Of the 645 samples tested, 208 (32.2%) tested were positive for NoV by EIA, among which 95 (45.7%) were genotyped. Among the genotyped samples, 12 (12.6%) were characterized as GII.17_2014 with the first case detected in November 2015 (1/30, 3.3%) and the others in 2016 (11/65, 16.9%). All strains found in our study were clustered in clade D (epidemic strain). The uncorrelated log-normal model estimations calculated the rate of evolution for GII-17 strains as 1.95 × 10- 3 (1.28 × 10- 3-2.63 × 10- 3). In total, 36 nucleotide changes were observed after analyzing the VP1 sequence, among which 28 occurred in the P2 region. CONCLUSIONS: These data demonstrate the evolutionary dynamics in NoV GII.17_2014 strains, which indicated high mutation rates with nucleotide substitutions and indels that are related to the elevated levels of antigenic diversity. This partly explains the increase in viral prevalence.


Asunto(s)
Infecciones por Caliciviridae/virología , Evolución Molecular , Gastroenteritis/virología , Tipificación Molecular , Norovirus/clasificación , Norovirus/genética , Brasil/epidemiología , Infecciones por Caliciviridae/epidemiología , Proteínas de la Cápside/genética , Enfermedades Transmisibles Emergentes/epidemiología , Enfermedades Transmisibles Emergentes/virología , Brotes de Enfermedades , Epidemias , Heces/virología , Gastroenteritis/epidemiología , Genotipo , Humanos , Tipificación Molecular/métodos , Filogenia , Prevalencia , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Virología/métodos
8.
Nat Med ; 25(12): 1865-1872, 2019 12.
Artículo en Inglés | MEDLINE | ID: mdl-31792456

RESUMEN

Viruses are implicated in autoimmune destruction of pancreatic islet ß cells, which results in insulin deficiency and type 1 diabetes (T1D)1-4. Certain enteroviruses can infect ß cells in vitro5, have been detected in the pancreatic islets of patients with T1D6 and have shown an association with T1D in meta-analyses4. However, establishing consistency in findings across studies has proven difficult. Obstacles to convincingly linking RNA viruses to islet autoimmunity may be attributed to rapid viral mutation rates, the cyclical periodicity of viruses7 and the selection of variants with altered pathogenicity and ability to spread in populations. ß cells strongly express cell-surface coxsackie and adenovirus receptor (CXADR) genes, which can facilitate enterovirus infection8. Studies of human pancreata and cultured islets have shown significant variation in enteroviral virulence to ß cells between serotypes and within the same serotype9,10. In this large-scale study of known eukaryotic DNA and RNA viruses in stools from children, we evaluated fecally shed viruses in relation to islet autoimmunity and T1D. This study showed that prolonged enterovirus B rather than independent, short-duration enterovirus B infections may be involved in the development of islet autoimmunity, but not T1D, in some young children. Furthermore, we found that fewer early-life human mastadenovirus C infections, as well as CXADR rs6517774, independently correlated with islet autoimmunity.


Asunto(s)
Autoinmunidad/inmunología , Diabetes Mellitus Tipo 1/virología , Enterovirus/aislamiento & purificación , ARN Viral/aislamiento & purificación , Adolescente , Autoinmunidad/genética , Niño , Preescolar , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 1/inmunología , Diabetes Mellitus Tipo 1/patología , Enterovirus/inmunología , Enterovirus/patogenicidad , Heces/virología , Femenino , Humanos , Lactante , Insulina/metabolismo , Células Secretoras de Insulina/inmunología , Células Secretoras de Insulina/virología , Islotes Pancreáticos/inmunología , Islotes Pancreáticos/patología , Islotes Pancreáticos/virología , Masculino , Páncreas/inmunología , Páncreas/patología , Páncreas/virología
9.
BMC Infect Dis ; 19(1): 978, 2019 Nov 21.
Artículo en Inglés | MEDLINE | ID: mdl-31752744

RESUMEN

BACKGROUND: Acute diarrhea is a leading cause of morbidity and mortality in children particularly in developing countries of Asia and Africa. The present study was conducted to detect the two most important pathogens, rotavirus and Campylobacter Jejuni in children suffering with diarrhea in Rawalpindi and Islamabad, Pakistan in 2014. The clinical and epidemiological aspects of the disease were also investigated. METHODS: A total of 500 stool samples were collected from children presented with clinical signs and symptoms of acute diarrhea. The samples were initially screened for the presence of rotavirus A (RVA) via ELISA (Enzyme-linked immunosorbent assay) and RT-PCR (Reverse Transcriptase PCR) and then were analysed for C. jejuni by using species specific PCR assay. RESULTS: The detection rate of RVA was 26.4% (132/500) while, Campylobacter was detected in 52% (260/500) of samples with C. jejuni accounted for 48.2% (241/500) of all study cases. Co-infection of C. jejuni with RVA was identified in 21.8% of all cases. Children with RVA and C. jejuni co-infection showed a higher probability (p = 0.01) to be dehydrated. A significant association (p = 0.02) was found between C. jejuni positive status and fever in children. The median age of children with both RVA and C. jejuni infection was 6-11 months. The RVA detection rate was high in winter months of the year while, C. jejuni infections were documented high in summer over 1 year study period. CONCLUSIONS: The overall results have demonstrated the high prevalence of C. jejuni in Rawalpindi, Islamabad, Pakistan in 2014. The results of present study will not only help to calculate disease burden caused by C. jejuni and rotavirus but also will provide critical information to health authorities in planning public health care strategies against these pathogens.


Asunto(s)
Infecciones por Campylobacter/microbiología , Campylobacter jejuni/aislamiento & purificación , Diarrea/microbiología , Diarrea/virología , Infecciones por Rotavirus/virología , Rotavirus/aislamiento & purificación , Infecciones por Campylobacter/epidemiología , Campylobacter jejuni/clasificación , Campylobacter jejuni/genética , Preescolar , Ciudades , Coinfección/epidemiología , Coinfección/microbiología , Coinfección/virología , Diarrea/epidemiología , Heces/microbiología , Heces/virología , Femenino , Humanos , Lactante , Masculino , Pakistán/epidemiología , Prevalencia , Rotavirus/clasificación , Rotavirus/genética , Infecciones por Rotavirus/epidemiología
10.
Arch Virol ; 164(12): 3151-3155, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31616994

RESUMEN

Astroviruses (AstVs) have a very wide range of hosts and are associated with enteric and extra-enteric disease in mammals and birds. Cross-species transmission of AstVs has been observed frequently. In the present study, the genome of a novel astrovirus from Amur tigers (Panthera tigris) from a zoo in China was characterized and was found to have the typical genomic features of other mammal AstVs. It showed the highest nucleotide sequence similarity (46.1-87.3% identity) to AstVs from cats, indicating a close phylogenetic relationship and possible cross-species transmission between them. To our knowledge, this is the first identification and characterization of AstV from tigers, and this virus is the third astrovirus identified in hosts of the family Felidae. The results of this study will be helpful for understanding the origin, genetic diversity, and cross-species transmission of AstV.


Asunto(s)
Animales de Zoológico/virología , Infecciones por Astroviridae/veterinaria , Astroviridae/aislamiento & purificación , Tigres/virología , Animales , Astroviridae/clasificación , Astroviridae/genética , Infecciones por Astroviridae/virología , Gatos , China , Heces/virología , Filogenia , Análisis de Secuencia de ADN
11.
BMC Public Health ; 19(1): 1340, 2019 Oct 22.
Artículo en Inglés | MEDLINE | ID: mdl-31640646

RESUMEN

BACKGROUND: Diarrhea is a major cause of morbidity and mortality, yet incidence and etiology data are limited. We conducted laboratory-based diarrhea surveillance in Guatemala. METHODS: A diarrhea case was defined as ≥3 loose stools in a 24-h period in a person presenting to the surveillance facilities. Epidemiologic data and stool specimens were collected. Specimens were tested for bacterial, parasitic, and viral pathogens. Yearly incidence was adjusted for healthcare seeking behaviors determined from a household survey conducted in the surveillance catchment area. RESULTS: From November 2008 to December 2012, the surveillance system captured 5331 diarrhea cases; among these 1381 (26%) had specimens tested for all enteric pathogens of interest. The adjusted incidence averaged 659 diarrhea cases per 10,000 persons per year, and was highest among children aged < 5 years, averaging 1584 cases per 10,000 children per year. Among 1381 (26%) specimens tested for all the pathogens of interest, 235 (17%) had a viral etiology, 275 (20%) had a bacterial, 50 (4%) had parasites, and 86 (6%) had co-infections. Among 827 (60%) specimens from children aged < 5 years, a virus was identified in 196 (23%) patients; 165 (20%) had norovirus and 99 (12%) rotavirus, including co-infections. Among 554 patients aged ≥5 years, 103 (19%) had a bacterial etiology, including diarrheagenic Escherichia coli in 94 (17%) cases, Shigella spp. in 31 (6%), Campylobacter spp. in 5 (1%), and Salmonella spp. in 4 (1%) cases. Detection of Giardia and Cryptosporidium was infrequent (73 cases; 5%). CONCLUSIONS: There was a substantial burden of viral and bacterial diarrheal diseases in Guatemala, highlighting the importance of strengthening laboratory capacity for rapid detection and control and for evaluation of public health interventions.


Asunto(s)
Disentería/epidemiología , Disentería/etiología , Vigilancia en Salud Pública/métodos , Adolescente , Adulto , Niño , Preescolar , Heces/microbiología , Heces/parasitología , Heces/virología , Femenino , Guatemala/epidemiología , Humanos , Incidencia , Lactante , Laboratorios , Masculino , Persona de Mediana Edad , Adulto Joven
12.
Arch Virol ; 164(12): 3157-3160, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31641840

RESUMEN

Here, we report a novel virulent P2-like bacteriophage, R18C, isolated from rabbit faeces, which, in addition to Escherichia coli K-12 strains, was able to be propagated on Citrobacter rodentium strain ICC169 and a range of Shigella sonnei strains with high efficiency of plating (EOP). It represents the first lytic bacteriophage originating from rabbit and the first infectious P2-like phage of animal origin. In the three characteristic moron-containing regions of P2-like phages, R18C contains genes with unknown function that have so far only been found in cryptic P2-like prophages.


Asunto(s)
Bacteriófagos/aislamiento & purificación , Citrobacter rodentium/virología , Conejos/microbiología , Shigella sonnei/virología , Animales , Bacteriófagos/clasificación , Bacteriófagos/genética , Citrobacter rodentium/fisiología , Heces/virología , Genoma Viral , Profagos/clasificación , Profagos/genética , Profagos/aislamiento & purificación , Shigella sonnei/fisiología
13.
BMC Infect Dis ; 19(1): 910, 2019 Oct 29.
Artículo en Inglés | MEDLINE | ID: mdl-31664944

RESUMEN

BACKGROUND: On September 4, 2018, a boarding school in the Shunyi District of Beijing, China reported an outbreak of acute gastroenteritis. At least 209 suspected students caused of diarrhea and vomiting. The case was investigated, and control measures were taken to prevent further spread. METHODS: A retrospective cohort study was conducted among the school students and staff in order to test hypothesis that high risk of food served at the school canteen. We collected information on demographics, refectory records, person to person transmission by uniform epidemiological questionnaire. Risk ratios (RR) and 95% confidence intervals (CI) were calculated. Stool specimens of cases and canteen employees, retained food, water, and environmental swabs were investigated by laboratory analysis. RESULTS: We identified 209 cases (including 28 laboratory-confirmed cases) which occurred from August 29 to September 10. All cases were students, and the average age was 20, 52% were male. The outbreak lasted for 13 days, and peaked on September 5. Consumption of Drinks stall and Rice flour stall on September 1 (RR:3.4, 95%CI:1.5-7.8, and RR:7.6, 95%CI:2.8-20.2), Rice flour stall and Fish meal stall on September 2 (RR:4.0, 95%CI:1.2-13.6, and RR:4.6, 95%CI:1.7-12.5), muslim meal stall on September 4 (RR:2.7, 95%CI:1.3-5.4), Barbeque stall on September 5 (RR:3.0, 95%CI:1.2-7.0) were independently associated with increased risk of disease within the following 2 days. Among 35 specimens of rectal swabs or feces from students, 28 specimens were positive. Norovirus GI.6 alone was detected in 23 specimens, Bacillus cereus alone in 3 specimens and both norovirus GI.6 and Bacillus cereus in 2 specimens. Ten specimens of rectal swabs from canteen employees were positive for norovirus GI, and 2 specimens were positive for Bacillus cereus. Four retained food specimens were positive for Bacillus cereus, and environmental samples were negative for any viruses or bacteria. CONCLUSION: Our investigation indicated that canteen employees were infected by two pathogens (norovirus and Bacillus cereus) and transmission may have been possible due to unhygienic practices. Student consumption of food or drink at high-risk stalls was determined as the probable cause of the outbreak.


Asunto(s)
Bacillus cereus/aislamiento & purificación , Infecciones por Caliciviridae/epidemiología , Brotes de Enfermedades , Enfermedades Transmitidas por los Alimentos/microbiología , Gastroenteritis/epidemiología , Norovirus/aislamiento & purificación , Adolescente , Beijing/epidemiología , Infecciones por Caliciviridae/complicaciones , Diarrea/complicaciones , Heces/microbiología , Heces/virología , Femenino , Contaminación de Alimentos , Enfermedades Transmitidas por los Alimentos/virología , Gastroenteritis/complicaciones , Higiene de las Manos , Humanos , Masculino , Oportunidad Relativa , Estudios Retrospectivos , Encuestas y Cuestionarios , Universidades , Vómitos/complicaciones , Adulto Joven
14.
Vet Ital ; 55(3): 221-229, 2019 Sep 30.
Artículo en Inglés | MEDLINE | ID: mdl-31599546

RESUMEN

Porcine astroviruses (PoAstV) are found in the gastrointestinal tract of healthy and diseased pigs worldwide. However, their role in causing enteric disease in pigs and other animals has not been elucidated. In the present report, we describe for the first time in Italy, the identification and genetic characterization, through whole genome sequencing, of a PoAstV2 in pigs in Northeast Italy in 2015. This instance is the first detection of PoAstV2 in pigs in Italy. The phylogenetic analysis of the complete ORF2 segment highlights the high similarity of this virus to those circulating that same year in Japan. There are very few full astrovirus genomes available, and the present data represent an important contribution towards a better understanding of the characteristics and evolution of these viruses.


Asunto(s)
Infecciones por Astroviridae/veterinaria , Genoma Viral , Mamastrovirus/genética , Enfermedades de los Porcinos/virología , Animales , Infecciones por Astroviridae/virología , Heces/virología , Femenino , Italia , Mamastrovirus/clasificación , Filogenia , Análisis de Secuencia de ARN/veterinaria , Sus scrofa , Porcinos , Secuenciación Completa del Genoma/veterinaria
15.
Wei Sheng Yan Jiu ; 48(4): 545-559, 2019 Jul.
Artículo en Chino | MEDLINE | ID: mdl-31601334

RESUMEN

OBJECTIVE: Real-time reverse transcription-polymerase chain reaction(real-time RT-PCR) assay based on Taqman and phylogenetic tree were developed for detecting hepatitis E virus in swine feces of pig farms from several provinces and city. METHODS: Designing prime and probe refering to HEV genotype sequences of Genbank, we developed a Taqman-based real-time RT-PCR assay and nested RT-PCR according to HEV conserved domain after optimizing reaction system, then detected the prevalence of HEV infection of pig farms. RESULTS: The sensitivity of the real-time RT-PCR assay established in this experiment was 19. 9 copies/µL, the amplification efficiency was 92. 9%-109. 1%, there was no cross reaction with Sapovirus, Norovirus and Hepatitis A. A total of 342 samples of swine feces were detected. There were two hundred and ten positive samples, and positive rate was 61. 4%. The positive rate of before-fattening was 56. 6%, and after-fattening was 66. 9%. The positive rate of before and after fattening samples had statistical difference(χ~2=24. 8, P<0. 05). The genotype identification system determined that the positive strains isolated in this study were HEV-4 type, and three subtypes of 4 b, 4 d and 4 h were detected. CONCLUSION: The pig farms of several provinces and city are contaminated by HEV extensively. The genotypes of the isolated strains are all HEV-4 type. The infection rate and infection subtype of pigs in different provinces and cities are different.


Asunto(s)
Heces/virología , Virus de la Hepatitis E , Epidemiología Molecular , Animales , Genotipo , Hepatitis E , Filogenia , ARN Viral , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transcripción Reversa , Porcinos , Enfermedades de los Porcinos/virología
16.
Arch Virol ; 164(11): 2761-2768, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31506786

RESUMEN

A multiplex polymerase chain reaction (mPCR) assay was developed to detect and distinguish feline panleukopenia virus (FPV), feline bocavirus (FBoV) and feline astrovirus (FeAstV). Three pairs of primers were designed based on conserved regions in the genomic sequences of the three viruses and were used to specifically amplify targeted fragments of 237 bp from the VP2 gene of FPV, 465 bp from the NP1 gene of FBoV and 645 bp from the RdRp gene of FeAstV. The results showed that this mPCR assay was effective, because it could detect at least 2.25-4.04 × 104 copies of genomic DNA of the three viruses per µl, was highly specific, and had a good broad-spectrum ability to detect different genotypes of the targeted viruses. A total of 197 faecal samples that had been screened previously for FeAstV and FBoV were collected from domestic cats in northeast China and were tested for the three viruses using the newly developed mPCR assay. The total positive rate for these three viruses was 59.89% (118/197). From these samples, DNA from FPV, FBoV and FeAstV was detected in 73, 51 and 46 faecal samples, respectively. The mPCR testing results agreed with the routine PCR results with a coincidence rate of 100%. The results of this study show that this mPCR assay can simultaneously detect and differentiate FPV, FBoV and FeAstV and can be used as an easy, specific and efficient detection tool for clinical diagnosis and epidemiological investigation of these three viruses.


Asunto(s)
Bocavirus/genética , Proteínas de la Cápside/genética , Virus de la Panleucopenia Felina/genética , Mamastrovirus/genética , Reacción en Cadena de la Polimerasa Multiplex/métodos , Animales , Bocavirus/aislamiento & purificación , Enfermedades de los Gatos/diagnóstico , Enfermedades de los Gatos/virología , Gatos , China , Cartilla de ADN/genética , Heces/virología , Virus de la Panleucopenia Felina/aislamiento & purificación , Mamastrovirus/aislamiento & purificación , Filogenia , Análisis de Secuencia de ADN
17.
Arch Virol ; 164(12): 3065-3071, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31549303

RESUMEN

Tombusviruses are generally considered plant viruses. A novel tombus-/carmotetravirus-like RNA virus was identified in a faecal sample and blood and muscle tissues from a wild northern white-breasted hedgehog (Erinaceus roumanicus). The complete genome of the virus, called H14-hedgehog/2015/HUN (GenBank accession number MN044446), is 4,118 nucleotides in length with a readthrough stop codon of type/group 1 in ORF1 and lacks a poly(A) tract at the 3' end. The predicted ORF1-RT (RdRp) and the capsid proteins had low (31-33%) amino acid sequence identity to unclassified tombus-/noda-like viruses (Hubei tombus-like virus 12 and Beihai noda-like virus 10), respectively, discovered recently in invertebrate animals. An in vivo experimental plant inoculation study showed that an in vitro-transcribed H14-hedgehog/2015/HUN viral RNA did not replicate in Nicotiana benthamiana, Chenopodium quinoa, or Chenopodium murale, the most susceptible hosts for plant-origin tombusviruses.


Asunto(s)
Erizos/virología , Análisis de Secuencia de ARN/métodos , Tombusvirus/clasificación , Animales , Heces/virología , Tamaño del Genoma , Genoma Viral , Especificidad del Huésped , Músculos/virología , Filogenia , Tombusvirus/genética , Tombusvirus/aislamiento & purificación
18.
Intervirology ; 62(3-4): 164-168, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31487720

RESUMEN

Despite the significant medical advances which have taken place in the last decades, acute diarrhoea cases remain a public health issue of major significance, with gastroenteritis agents being associated with severe symptoms in adults and high morbidity in infants and children. Regarding rotaviruses, while children are the predominant victims of rotavirus infection, adults (often caretakers or parents of these children) may experience the same symptoms of fever, vomiting, and non-bloody diarrhoea. Three different routine schemes for the detection of rotaviruses in archived stool samples were evaluated in terms of diagnostic performance. A total of 640 archived stool samples were included in the study. The samples were screened with three different techniques: a commercial rapid immunochromatographic test, a modified in-house conventional one-step reverse-transcription polymerase chain reaction (PCR) screen protocol, and a com-mer-cial one-step real-time PCR kit. Technical aspects and considerations are discussed.


Asunto(s)
Pruebas Diagnósticas de Rutina/métodos , Inmunoensayo/métodos , Infecciones por Rotavirus/diagnóstico , Rotavirus/aislamiento & purificación , Diarrea/diagnóstico , Diarrea/virología , Heces/virología , Gastroenteritis/diagnóstico , Gastroenteritis/virología , Grecia , Humanos , Técnicas de Diagnóstico Molecular/métodos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Rotavirus/inmunología , Sensibilidad y Especificidad , Centros de Atención Terciaria
19.
Zhonghua Liu Xing Bing Xue Za Zhi ; 40(8): 883-888, 2019 Aug 10.
Artículo en Chino | MEDLINE | ID: mdl-31484248

RESUMEN

Objective: To investigate the epidemiologic characteristics of Norovirus among adult patients suffering from infectious diarrhea in Shanghai, so as to provide evidence for developing related strategies on prevention and control of the disease. Methods: Diarrheal outpatients were monitored at the 'Intestinal clinic' from 22 hospitals involved in the sentinel surveillance program in Shanghai. Information on demographic and epidemiologic features of the patients was collected while data and clinical, fecal specimens were collected and sent to the district CDC for Norovirus detection. Positive rates of Norovirus were also compared in various populations and seasons during 2013-2018. Multivariate logistic regression model was adopted to fit into the comparisons between non-Norovirus and Norovirus groups. Results: 19.28% of the 12 083 diarrheal cases were found to have carried the Norovirus, with GⅡgroup the most commonly identified genotype. Rates of detection was seen higher in males (20.78%) than in females (17.73%). 30-44 year-old were found having the highest positive rate (21.51%). The positive rates were found the highest (23.60%) in the year of 2015. All the above shown differences were statistically significant (P<0.05). Diarrheal patients affected with Norovirus would present watery stool (75.94%) and vomiting (35.84%). Data from the multivariate logistic analysis showed that factors as: being males, 30-44 years old, officials/clerks, in winter season and with histories of travelling etc., were related to higher risks on Norovirus infection. Conclusions: Infectious diarrhea caused by Norovirus occurred all year round, with seasonal peaks seen in winter and spring, in Shanghai. Specific prevention and control measures should be taken on Norovirus- caused infectious diarrhea, according to the difference on age, gender and season.


Asunto(s)
Infecciones por Caliciviridae/epidemiología , Diarrea/epidemiología , Disentería/epidemiología , Heces/virología , Gastroenteritis/epidemiología , Norovirus/aislamiento & purificación , Adulto , Infecciones por Caliciviridae/diagnóstico , China/epidemiología , Diarrea/diagnóstico , Diarrea/virología , Disentería/diagnóstico , Disentería/virología , Femenino , Gastroenteritis/virología , Humanos , Masculino , Norovirus/genética , Vigilancia de Guardia
20.
Zhonghua Liu Xing Bing Xue Za Zhi ; 40(8): 889-894, 2019 Aug 10.
Artículo en Chino | MEDLINE | ID: mdl-31484249

RESUMEN

Objective: To understand the seasonality and etiological characteristics of infectious diarrhea in adults from Shanghai. Methods: Adult patients with diarrhea who had visited the enteric disease clinics in 22 hospitals that carrying on the Diarrhea Comprehensive Surveillance sentinel programs in Shanghai during 2014-2017, were surveyed. Stool specimens were collected according to the different intervals of sampling and detected for 12 bacteria and 5 viruses. Concentration ratio and circular distribution method were used for data analysis. Results: From 2014 to 2017, a total of 9 573 stool specimens were collected from the targeted diarrhea patients ≥18 years old (n=96 067), through the Shanghai Diarrhea Comprehensive Surveillance program. The positive rate of detection was 46.44%. Seasonal peaks of infectious diarrhea were both seen in summer (bacteria peak, diarrheagenic Escherichia coli and Vibrio parahaemolyticus, etc.) and in winter (virus peak, Norovirus, etc.). Both bacterial and viral infections presented seasonal concentration (Raleigh's test P<0.001) but more obvious with bacterial infection. Viral infection accounted for 60.19% of the cause of infectious diarrhea. The top five predominant pathogens appeared as Norovirus, Rotavirus, diarrheagenic Escherichia coli, Vibrio parahaemolyticus, and Salmonella spp.. Conclusions: Among the adult outpatients with infectious diarrhea in Shanghai, obvious seasonality was seen, with peaks in both summer and winter. Viral infection with Norovirus in particular, appeared as the predominant source of infection. Active, continuous and comprehensive diarrhea-related surveillance programs would be able to monitor the changing dynamic of pathogen spectrum, and lead to the adoption of targeted preventive measures.


Asunto(s)
Bacterias/aislamiento & purificación , Diarrea/diagnóstico , Diarrea/etiología , Disentería/diagnóstico , Disentería/etiología , Heces , Pacientes Ambulatorios/estadística & datos numéricos , Vigilancia de la Población/métodos , Virus/aislamiento & purificación , Adolescente , Adulto , Bacterias/clasificación , Niño , Preescolar , China/epidemiología , Diarrea/epidemiología , Disentería/epidemiología , Escherichia coli/aislamiento & purificación , Heces/microbiología , Heces/virología , Humanos , Persona de Mediana Edad , Norovirus/aislamiento & purificación , Rotavirus/aislamiento & purificación , Salmonella/clasificación , Salmonella/aislamiento & purificación , Estaciones del Año , Vibrio parahaemolyticus/aislamiento & purificación , Virus/clasificación
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