RESUMO
Hepatitis E caused by hepatitis E virus (HEV) is considered an emerging foodborne zoonosis in industrialized, non-endemic countries. Domestic pigs and wild boars are considered the main reservoir of HEV. However, HEV can also infect an ever-expanding host range of animals, but they exact role in transmitting the virus to other species or humans is mostly unknown. In this work, we investigated the spread of HEV in free-living and captive spotted deer (Axis axis) from Uruguay in a 2-year period (2020-2022) and examined the role of this invasive species as a new potential reservoir of the virus. In addition, with the aim to gain new insights into viral ecology in the context of One Health, by using camera trapping, we identified and quantified temporal and spatial coexistence of spotted deer, wild boars, and cattle. In free-living animals, we detected an anti-HEV seropositivity of 11.1% (6/54). HEV infection and viral excretion in feces were assessed by RT-PCR. Thirteen of 19 samples (68.4%) had HEV RNA. Six samples were amplified using a broadly reactive RT-PCR and sequenced. No captive animal showed evidence of HEV infection. Additionally, HEV RNA was detected in a freshwater pond shared by these species. Phylogenetic and p-distance analysis revealed that zoonotic HEV genotype 3 strains circulate together with unclassified variants related to moose HEV whose potential risk of transmission to humans and other domestic and wild animals is unknown. The data presented here suggest that spotted deer (A. axis) may be a novel host for zoonotic HEV strains.
Assuntos
Cervos , Vírus da Hepatite E , Hepatite E , Suínos , Humanos , Bovinos , Animais , Vírus da Hepatite E/genética , Filogenia , Uruguai , Sus scrofa , RNA Viral/genética , GenótipoRESUMO
Hepatitis E virus (HEV) is an emerging zoonotic pathogen associated with relevant public health issues. The aim of this study was to investigate HEV presence in free-living capybaras inhabiting urban parks in São Paulo state, Brazil. Molecular characterization of HEV positive samples was undertaken to elucidate the genetic diversity of the virus in these animals. A total of 337 fecal samples were screened for HEV using RT-qPCR and further confirmed by conventional nested RT-PCR. HEV genotype and subtype were determined using Sanger and next-generation sequencing. HEV was detected in one specimen (0.3%) and assigned as HEV-3f. The IAL-HEV_921 HEV-3f strain showed a close relationship to European swine, wild boar and human strains (90.7-93.2% nt), suggesting an interspecies transmission. Molecular epidemiology of HEV is poorly investigated in Brazil; subtype 3f has been reported in swine. This is the first report of HEV detected in capybara stool samples worldwide.
Assuntos
Vírus da Hepatite E , Humanos , Animais , Suínos , Brasil/epidemiologia , Vírus da Hepatite E/genética , Roedores , Fezes , GenótipoRESUMO
Hepatitis E Virus (HEV) is an etiologic agent of hepatitis worldwide. HEV genotype 3 is the most prevalent in non-endemic regions, identified in humans, pigs and environmental samples. Thus, considering the zoonotic nature of HEV genotype 3, viral genome detection in wastewater concerns public health authorities. Electrochemical biosensors are promising analytical tools for viral genome detection in outside settings. This work reports on a highly specific, sensitive and portable electrochemical genosensor to detect HEV genotype 3 in wastewater samples. Based on the alignment analysis of HEV genotype 3 genome sequences available in GenBank, highly specific DNA target probes were designed to hybridize a target sequence within the ORF2/ORF3 overlapping genome region of HEV in between a biotinylated capture probe and a signal probe labeled with digoxigenin, in a sandwich-type format. An anti-Dig antibody labeled with the horseradish peroxidase (HRP) enzyme allowed electrochemical detection. The specificity of the target molecular probes of the viral genome was determined before the biosensor assembly by in silico analysis, PCR and qPCR assays demonstrating efficient amplification of two targets, i.e., nucleotides 5338-5373 and 5328-5373, but this last one of higher performance. The electrochemical response of the genosensor with synthetic HEV was target concentration-dependent in a linear range from 300 pM to 2.4 nM, with a sensitivity of 16.93 µA/nM, a LOD 1.2 pM and high reproducibility. The genosensor response was differential when interrogated with the HEV genotype 3 viral genomes from wastewater against other four viruses. Therefore, the approach offers a step forward to the epidemiologic surveillance of viruses in wastewater as an early warning system.
Assuntos
Vírus da Hepatite E , Hepatite E , Animais , Genótipo , Hepatite E/epidemiologia , Vírus da Hepatite E/genética , Humanos , Reprodutibilidade dos Testes , Suínos , Águas ResiduáriasRESUMO
BACKGROUND: Few studies about the evolutionary history of the hepatitis E virus (HEV) have been conducted. The aim of our work was to investigate and make inferences about the origin and routes of dispersion of HEV-3 in Argentina. METHODS: Phylogenetic, coalescent and phylogeographic analyses were performed using a 322-bp ORF2 genomic fragment of all HEV-3 sequences with known date and place of isolation published at GenBank until May 2018 (n=926), including 16 Argentinian sequences (isolated from pigs, water and humans). RESULTS: Phylogenetic analysis revealed two clades within HEV-3: abchij and efg. All Argentinian samples were grouped intermingled within clade 3abchij. The coalescent analysis showed that the most recent common ancestor for the clade 3abchij would have existed around the year 1967 (95% highest posterior density (HPD): 1963-1970). The estimated substitution rate was 1.01×10-2 (95%HPD: 9.3×10-3-1.09×10-2) substitutions/site/y, comparable with the rate previously described. The phylogeographic approach revealed a correspondence between phylogeny and place of origin for Argentinian samples, suggesting many HEV introductions in the country, probably from Europe and Japan. CONCLUSIONS: This is the first evolutionary inference of HEV-3 that includes Argentinian strains, showing the circulation of many HEV-3 subtypes, obtained from different sources and places, with recent diversification processes. ACCESSION NUMBERS: [KX812460], [KX812461], [KX812462], [KX812465], [KX812466], [KX812467], [KX812468], [KX812469].
Assuntos
Vírus da Hepatite E , Hepatite E , Animais , Argentina/epidemiologia , Genótipo , Hepatite E/epidemiologia , Humanos , Filogenia , Filogeografia , SuínosRESUMO
Patients with hepatitis C virus (HCV) genotype 3 (GT3) infection are resistant to direct-acting antiviral (DAA) treatments. This study aimed to analyze the effectiveness of sofosbuvir (SOF)+daclatasvir (DCV) ± ribavirin (RBV); SOF+velpatasvir (VEL)±RBV; SOF+VEL+voxilaprevir (VOX); and glecaprevir (GLE)+pibrentasvir (PIB) in the treatment of HCV GT3-infected patients in real-world studies. Articles were identified by searching the PubMed, EMBASE, and Cochrane Library databases from January 1, 2016 to September 10, 2019. The meta-analysis was conducted to determine the sustained virologic response (SVR) rate, using R 3.6.2 software. Thirty-four studies, conducted on a total of 7328 patients from 22 countries, met the inclusion criteria. The pooled SVR rate after 12/24 weeks of treatment was 92.07% (95% CI: 90.39-93.61%) for the evaluated regimens. Also, the SVR rate was 91.17% (95% CI: 89.23-92.94%) in patients treated with SOF+DCV±RBV; 95.08% (95% CI: 90.88-98.13%) in patients treated with SOF+VEL±RBV; 84.97% (95% CI: 73.32-93.91%) in patients treated with SOF+VEL+VOX; and 98.54% (95% CI: 96.40-99.82%) in patients treated with GLE+PIB. The pooled SVR rate of the four regimens was 95.24% (95% CI: 93.50-96.75%) in non-cirrhotic patients and 89.39% (95% CI: 86.07-92.33%) in cirrhotic patients. The pooled SVR rate was 94.41% (95% CI: 92.02-96.42%) in treatment-naive patients and 87.98% (95% CI: 84.31-91.25%) in treatment-experienced patients. The SVR rate of GLE+PIB was higher than other regimens. SOF+VEL+VOX can be used as a treatment regimen following DAA treatment failure.
Assuntos
Antivirais/uso terapêutico , Benzimidazóis/uso terapêutico , Carbamatos/uso terapêutico , Hepatite C/tratamento farmacológico , Compostos Heterocíclicos de 4 ou mais Anéis/uso terapêutico , Imidazóis/uso terapêutico , Compostos Macrocíclicos/uso terapêutico , Pirrolidinas/uso terapêutico , Quinoxalinas/uso terapêutico , Sofosbuvir/uso terapêutico , Sulfonamidas/uso terapêutico , Valina/análogos & derivados , Combinação de Medicamentos , Hepatite C/virologia , Humanos , Ribavirina/uso terapêutico , Valina/uso terapêuticoRESUMO
Hepatitis C virus (HCV) genotype 3 presents a high level of both baseline and acquired resistance to direct-acting antivirals (DAAs), particularly those targeting the NS5A protein. To understand this resistance we studied a cohort of Brazilian patients treated with the NS5A DAA, daclatasvir and the nucleoside analogue, sofosbuvir. We observed a novel substitution at NS5A amino acid residue 98 [serine to glycine (S98G)] in patients who relapsed post-treatment. The effect of this substitution on both replication fitness and resistance to DAAs was evaluated using two genotype 3 subgenomic replicons. S98G had a modest effect on replication, but in combination with the previously characterized resistance-associated substitution (RAS), Y93H, resulted in a significant increase in daclatasvir resistance. This result suggests that combinations of substitutions may drive a high level of DAA resistance and provide some clues to the mechanism of action of the NS5A-targeting DAAs.
Assuntos
Antivirais/farmacologia , Carbamatos/farmacologia , Farmacorresistência Viral/genética , Hepacivirus/efeitos dos fármacos , Imidazóis/farmacologia , Pirrolidinas/farmacologia , Valina/análogos & derivados , Proteínas não Estruturais Virais/genética , Antivirais/uso terapêutico , Brasil , Carbamatos/uso terapêutico , Linhagem Celular Tumoral , Estudos de Coortes , Farmacorresistência Viral/efeitos dos fármacos , Quimioterapia Combinada , Genótipo , Hepacivirus/genética , Hepacivirus/fisiologia , Hepatite C/tratamento farmacológico , Hepatite C/virologia , Humanos , Imidazóis/uso terapêutico , Mutação , Pirrolidinas/uso terapêutico , Recidiva , Sofosbuvir/farmacologia , Sofosbuvir/uso terapêutico , Valina/farmacologia , Valina/uso terapêutico , Proteínas não Estruturais Virais/antagonistas & inibidores , Replicação Viral/genéticaRESUMO
Nowadays, the retreatment of patients with Hepatitis C virus (HCV) genotype 3 (GT3) especially cirrhotic, who have already been treated with regimens containing a NS5A inhibitor represents a challenge. Use a novel retreatment option for patients with a difficult approach. We present three case reports of retreatment with a new combination of Direct-acting antivirals (DAAs), Sofosbuvir, Elbasvir/Grazoprevir in patients with GT3 with a previous failure with Sofosbuvir/Ledipasvir. All the cases achieved sustained virologic response (SVR) at week +12 without adverse effects. In our experience, this combo may represent an effective and safe option for these patients.
Assuntos
Antivirais/uso terapêutico , Farmacorresistência Viral , Hepacivirus/genética , Hepatite C Crônica/tratamento farmacológico , RNA Viral/genética , Amidas , Benzofuranos/uso terapêutico , Carbamatos , Ciclopropanos , Quimioterapia Combinada , Genótipo , Hepatite C Crônica/virologia , Humanos , Imidazóis/uso terapêutico , Masculino , Pessoa de Meia-Idade , Quinoxalinas/uso terapêutico , Retratamento , Sofosbuvir/uso terapêutico , Sulfonamidas , Falha de TratamentoRESUMO
Hepatitis E virus (HEV), an emerging virus associated with acute hepatic disease, leads to thousands of deaths worldwide. HEV has already been reported in Brazil; however, there is a lack of epidemiological and molecular information on the genetic variability, taxonomy, and evolution of HEV. It is thus unclear whether hepatitis E is a neglected disease in Brazil or it has low relevance for public health in this country. Here, for the first time, we report the presence of HEV in Northeast Brazil. A total of 119 swine faecal samples were screened for the presence of HEV RNA using real-time polymerase chain reaction (RT-PCR) and further confirmed by conventional RT-PCR; among these, two samples were identified as positive. Molecular evolution analyses based on capsid sequences revealed that the samples had close proximities to HEV sequences belonging to genotype 3 and were genetically related to subtype 3f isolated in humans. Parsimony ancestral states analysis indicated gene flow events from HEV cross-species infection, suggesting an important role of pig hosts in viral spillover. HEV's ability for zoonotic transmission by inter-species host switching as well as its possible adaptation to new animal species remain important issues for human health.
Assuntos
Humanos , Animais , Zoonoses/virologia , Vírus da Hepatite E/isolamento & purificação , Vírus da Hepatite E/genética , Fezes/virologia , Filogenia , Suínos , Doenças dos Suínos/transmissão , Brasil , RNA Viral , Capsídeo/virologia , Hepatite E/virologia , Análise de Sequência de DNA , Reação em Cadeia da Polimerase em Tempo Real , GenótipoRESUMO
Hepatitis E is a zoonotic disease, recognized as an important global public health concern. In this study, molecular detection of the ORF1 and ORF2 genomic regions of the hepatitis E virus (HEV) was carried out in fecal and serum samples from pigs in subsistence farms of Mato Grosso, Brazil. Fragments of the ORF2 region were amplified in 8% (12/150) of fecal samples, with 53.3% (8/15) of farms having positive results. Of the 12 positive samples, fragments of the ORF1 region were amplified in 33.3% (4/12) of these. Molecular characterization confirmed the phylogenetic groupings as HEV subtypes 3d, 3â¯h, and 3i. The results revealed that meat from pigs that was originally meant for personal consumption is being traded in marketplaces in metropolitan Cuiabá, thereby creating a source of transmission to consumers in Mato Grosso. The environmental conditions must be taken into account when investigating the presence and transmission of HEV.
Assuntos
Genótipo , Vírus da Hepatite E/isolamento & purificação , Hepatite E/veterinária , Doenças dos Suínos/diagnóstico , Zoonoses/epidemiologia , Animais , Brasil/epidemiologia , Fazendas , Fezes/virologia , Variação Genética , Hepatite E/diagnóstico , Hepatite E/epidemiologia , Hepatite E/transmissão , Vírus da Hepatite E/classificação , Vírus da Hepatite E/genética , Carne/virologia , Filogenia , RNA Viral/genética , Suínos , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/transmissão , Doenças dos Suínos/virologia , Zoonoses/diagnóstico , Zoonoses/transmissão , Zoonoses/virologiaRESUMO
Directly-acting antivirals (DAA) have changed the chronic hepatitis C virus (HCV) infection therapeutic scenario allowing virus eradication in more than 95% of patients, independently from the genotype, with 12 to 24-week treatment regimens. We describe a 51-year-old Pakistani man with a chronic HCV-genotype 3 (GT3a) infection with moderate liver fibrosis, who achieved sustained virological response (SVR) 24 after a tripled dose of Daclatasvir (DCV) taken erroneously associated to Sofosbuvir (SOF). The patient had a concomitant intestinal TB infection whose treatment had been delayed in order to firstly eradicate HCV to reduce the liver toxicity of anti-mycobacterial drugs. Thanks to the cultural mediator support, we explained to the patient the correct posology of each drug to take during the day consisting of 12 week SOF (400 mg daily) plus DCV (60 mg daily) regimen. He returned 13 days after for a programmed visit and we were surprised to learn that he had taken 3 pills of DCV (180 mg/daily) instead of one, thus ending DCV assumption after only 9 days while SOF was taken correctly. He complained no symptoms. We immediately performed blood test that showed alteration of lactate dehydrogenase, creatine phosphokinase, and creatin kinase MB activity. At day 15 we stopped SOF closely monitoring the patient. Blood test alterations returned normal after one week of treatment suspension, HCV viremia remained suppressed after 4, 12 and 24 weeks proving HCV eradication. If confirmed, these data could suggest that higher doses of DCV, if tolerated, might be employed in short-time HCV-GT3 treatment.