RESUMEN
Following the March 2013 outbreak of novel avian influenza A(H7N9) virus in humans and the subsequent isolation of the virus from chickens, ducks and pigeons in the People's Republic of China, concerns were raised that the H7N9 virus would spread beyond China through the poultry value chain linking to a number of bordering countries. For this reason, a rapid emergency surveillance exercise took place in Bhutan between May and July 2013 with the objective of determining whether influenza A(H7N9) virus was silently circulating in domestic poultryandwild birds in Bhutan.Atotal of 1716 oropharyngeal,tracheal and cloacal swabs together with faecal droppings were collected from poultry, wild birds and feral pigeons throughout the country; these samples included 150 that had been previously collected for surveillance of influenza A(H5N1) virus. Overall, 733 of the samples were tested. A QIAamp Viral RNA Mini K it was used to extract viral RNA from a mix of oropharyngeal, tracheal and cloacal swabs and faecal droppings. The matrix gene of avian influenza type A virus was detected using a specific real-time quantitative reverse-transcription polymerase chain reaction (RT-PCR) assay, and positive samples were further tested in RT-PCR for simultaneous detection of the H7 and N9 genes. Among the 733 samples tested, 46 (26 prospective, 20 retrospective) were confirmed positive for influenza A, a prevalence of 6.3% (95% CI: 4.6 to 8.3). The influenza A-positive samples were from areas in the south of Bhutan that had experienced previous outbreaks of highly pathogenic influenza A(H5N1). None of the samples tested positive for H7N9 strains, providing evidence that influenza A(H7N9) virus was not present in the sampled population. A risk-based approach for surveillance of influenza A(H7N9) and H5N1 is recommended in Bhutan, based on the epidemiology of the disease in China and other countries in South and Southeast Asia.
Asunto(s)
Brotes de Enfermedades/veterinaria , Subtipo H7N9 del Virus de la Influenza A , Gripe Aviar/virología , Animales , Bután/epidemiología , Aves , Subtipo H5N1 del Virus de la Influenza A , Gripe Aviar/epidemiología , Vigilancia de la PoblaciónRESUMEN
Edwardsiella ictaluri is a Gram-negative facultative intracellular pathogen causing enteric septicemia of catfish (ESC). We have shown recently that tricarboxylic acid cycle (TCA) and one-carbon (C1) metabolism are involved in E. ictaluri pathogenesis. However, the effect of multiple mutations in these pathways is unknown. Here, we report four novel E. ictaluri mutants carrying double gene mutations in TCA cycle (EiΔmdhΔsdhC, EiΔfrdAΔsdhC), C1 metabolism (EiΔglyAΔgcvP), and both TCA and C1 metabolism pathways (EiΔgcvPΔsdhC). In-frame gene deletions were constructed by allelic exchange and mutants' virulence and vaccine efficacy were evaluated using in vivo bioluminescence imaging (BLI) as well as end point mortality counts in catfish fingerlings. Results indicated that all the double gene mutants were attenuated compared to wild-type (wt) E. ictaluri. There was a 1.39-fold average reduction in bioluminescence, and hence bacterial numbers, from all the mutants except for EiΔfrdAΔsdhC at 144 h post-infection. Vaccination with mutants was very effective in protecting channel catfish against subsequent infection with virulent E. ictaluri 93-146 strain. In particular, immersion vaccination resulted in complete protection. Our results provide further evidence on the importance of TCA and C1 metabolism pathways in bacterial pathogenesis.
Asunto(s)
Vacunas Bacterianas/administración & dosificación , Edwardsiella ictaluri/genética , Edwardsiella ictaluri/inmunología , Infecciones por Enterobacteriaceae/veterinaria , Enfermedades de los Peces/microbiología , Ictaluridae , Redes y Vías Metabólicas/genética , Animales , Carbono/metabolismo , Ciclo del Ácido Cítrico/genética , Edwardsiella ictaluri/metabolismo , Edwardsiella ictaluri/patogenicidad , Infecciones por Enterobacteriaceae/microbiología , Infecciones por Enterobacteriaceae/prevención & control , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/prevención & control , Eliminación de Gen , Genotipo , Ictaluridae/inmunología , Ictaluridae/microbiología , Mutación , Vacunación/veterinaria , Virulencia/genéticaRESUMEN
We characterized Influenza A/H5N1 virus that caused the first outbreak of highly pathogenic avian influenza (HPAI) in chickens in Bhutan in 2010. The virus was highly virulent to chicken, killing them within two days of the experimental inoculation with an intravenous pathogenicity index (IVPI) of 2.88. For genetic and phylogenetic analyses, complete genome sequencing of 4 viral isolates was carried out. The isolates revealed multiple basic amino acids at their hemagglutinin (HA) cleavage site, similar to other "Qinghai-like" H5N1 isolates. The receptor-binding site of HA molecule contained avian-like amino acids ((222)Q and (224)G). The isolates also contained amino acid residue K at position 627 of the PB2 protein, and other markers in NS 1 and PB1 proteins, highlighting the risk to mammals. However, the isolates were sensitive to influenza drugs presently available in the market. The sequence analysis indicated that the Bhutan viruses shared 99.1-100% nucleotide homology in all the eight genes among themselves and 2010 chicken isolate from Bangladesh (A/chicken/Bangladesh/1151-11/2010) indicating common progenitor virus. The phylogenetic analysis indicated that the Bhutan isolates belonged to sub-clade 2.2.3 (EMA 3) and shared common progenitor virus with the 2010 Bangladesh virus. Based on the evidence of phylogeny and molecular markers, it could be concluded that the outbreaks in Bhutan and Bangladesh in 2010 were due to independent introductions of the virus probably through migratory birds.
Asunto(s)
Brotes de Enfermedades/veterinaria , Subtipo H5N1 del Virus de la Influenza A/clasificación , Subtipo H5N1 del Virus de la Influenza A/genética , Gripe Aviar/epidemiología , Gripe Aviar/virología , Filogenia , Animales , Antivirales/farmacología , Bangladesh/epidemiología , Secuencia de Bases , Bután/epidemiología , Pollos/virología , Subtipo H5N1 del Virus de la Influenza A/efectos de los fármacos , Subtipo H5N1 del Virus de la Influenza A/patogenicidad , Gripe Aviar/mortalidad , Pruebas de Sensibilidad Microbiana , Datos de Secuencia MolecularRESUMEN
AIMS: To identify livestock husbandry practices important for transmission of foot-and-mouth disease (FMD) in the herds and villages of four regions in Bhutan. To consider using this information to enhance the current prevention and control programme, a consideration arising from the failure to control FMD in spite of a control programme in place. METHODS: Between March and May 2009, 383 livestock farmers originating from 80 villages in four districts of Bhutan were interviewed, using a structured questionnaire, about the livestock management practices and incidence of FMD in their herds. Multivariable logistic regression was used to quantify the risk factors that predicted the outcome variable 'farmer-diagnosed FMD in Bhutan'. RESULTS: Sixty-two percent (49/79) of the villages and 87/355 (24%) of herds surveyed had at least one outbreak of FMD within the 5 years preceding the survey. The odds of having FMD in a herd increased substantially (OR=39.2; p<0.0001) when cattle mixed with herds from other nearby villages compared with those where mixing did not occur. Those cattle herds mixing with six or more other herds within the same village were 5.3 times (p<0.0001) more likely to have had FMD than those mixed with fewer than six herds. Farmers who fed kitchen waste to cattle were 14.1 times (p<0.0001), and those who sent their animals for grazing in the forest were 3.1 times (p=0.014), more likely to report FMD in their herds than those who did not. Farmers who kept their cattle always housed in a shed during the day (OR=0.033) or at night (OR=0.29) were less likely to report FMD than those who did not (p<0.04). CONCLUSIONS: Mixing of cattle at grazing areas was identified as a risk factor for FMD. This indicates that spread from infected herds and villages, through close contact, could be an important source of disease for non-infected herds in Bhutan. Therefore, quarantining of early cases in affected herds or villages could reduce the spread of disease within and between villages. This study also highlights the potential role of feeding kitchen waste to cattle as a risk factor for FMD. The findings from this study could be considered for strengthening of the FMD control programme in Bhutan.
Asunto(s)
Enfermedades de los Bovinos/epidemiología , Fiebre Aftosa/epidemiología , Crianza de Animales Domésticos , Animales , Bután/epidemiología , Bovinos , Vivienda para Animales , Modelos Logísticos , Oportunidad Relativa , Prevalencia , Factores de RiesgoRESUMEN
We report a molecular epidemiological study of rabies virus (RABV) strains circulating in animal populations in Bhutan, and investigate potential origins of these viruses. Twenty-three RABV isolates originating from dogs and other domestic animals were characterized by sequencing the partial nucleoprotein (N) gene (395 bp). Phylogenetic analysis was conducted and the Bhutanese isolates were compared with rabies viruses originating from other parts of the world. Phylogenetic analysis showed that Bhutanese isolates were highly similar and were closely related to Indian strains and South Asian Arctic-like-1 viruses. Our study suggests that the rabies viruses spreading in southern parts of Bhutan have originated from a common ancestor, perhaps from the Indian virus strain.