[show abstract][hide abstract] ABSTRACT: The cat gene, coding for chloramphenicol acetyltransferase has been reported for conferring the chloramphenicol resistance for Riemerella anatipestifer. Chloramphenicol acetyltransferases, however, are unable to inactivate florfenicol. In this study, 66 R. anatipestifer isolates were investigated for their susceptibility to chloramphenicol and florfenicol and the presence of floR gene. Results showed nine florfenicol intermediate or resistant R. anatipestifer isolates were all floR positive. The expression of floR gene in E. coli and inhibition studies with PAβN indicated that the floR gene was as an efflux pump conferring resistance to both chloramphenicol and florfenicol. Southern hybridization revealed the floR was located in the plasmid DNA of five isolates and in the chromosomal DNA of four isolates. Furthermore, two novel floR-carrying plasmids designated pRA0726 and pRA0846 were sequenced completely. pRA0726 was 11,704 bp in size with 10 putative open reading frames which included the floR, catB and bla(OXA-209) resistance genes. The most differences between sequences of pRA0846 and pRA0726 were the absence of a bla(OXA-209) gene and the deletion of 321 nucleotides of orf1 in pRA0846. Plasmid curing tests demonstrated that pRA0726 carried functional coding proteins for resistance to phenicol and β-lactam antimicrobials. To the best of our knowledge, this is the first report of presence of the floR and bla(OXA-209) resistance genes in R. anatipestifer.
[show abstract][hide abstract] ABSTRACT: H5N2 viruses were isolated from cloacal swab samples of apparently healthy chickens in Taiwan in 2003 and 2008 during surveillance of avian influenza. Each of the viruses was eradicated by stamping out. The official diagnosis report indicated that the Intravenous Pathogenicity Indexes (IVPIs) of the isolates were 0.00 and 0.89, respectively, indicating that these were low pathogenic strains, although the hemagglutinin of the strain isolated in 2008 (Taiwan08) had multibasic amino acid residues at the cleavage site (PQRKKR/G). In the present study, these H5N2 viruses were assessed for their intravenous and intranasal pathogenicity for chickens. It was examined whether Taiwan08 acquires pathogenicity through consecutive passages in chickens. Intravenous pathogenicity of Taiwan08 depended upon the age of the chickens used for the IVPI test; all of the eight-week-old chickens intravenously inoculated with Taiwan08 showed clinical signs but survived for ten days post inoculation (IVPI=0.68), whereas all the six-week-old chickens died (IVPI=1.86). Taiwan08-P8, which were passaged in chickens for eight times, killed all the eight-week-old chickens (IVPI=2.36). The four-week-old chickens died after intranasal inoculation of Taiwan08-P8, indicating that Taiwan08 must have become highly pathogenic during circulation in chicken flocks. These results emphasize the importance of a stamping out policy for avian influenza even if the IVPI of the causal virus is low.
Journal of Veterinary Medical Science 02/2011; 73(6):767-72. · 0.88 Impact Factor
[show abstract][hide abstract] ABSTRACT: Riemerella anatipestifer is a Gram-negative bacterium that can cause disease in a wide range of wild and domesticated birds, especially waterfowl. The presence of an antibiotic-resistance gene in R. anatipestifer has not yet been reported, indicating the need for investigation. In the present study, 40.5% of R. anatipestifer isolates were found to exhibit resistance to chloramphenicol, while 45.9% showed intermediate resistance and 13.5% were susceptible to chloramphenicol, an antibiotic that has been prohibited for use in food animals in Taiwan since 2003. The resistance gene was identified as the cat gene and cloned by library sequencing. The prevalence of the cat gene in Taiwanese R. anatipestifer isolates was 78.4%. The position of the cat gene was then determined within the novel plasmid, designated pRA0511. pRA0511 was sequenced and shown to be 11,435 bp in size with 10 open reading frames (ORFs). Proteins putatively encoded by these 10 ORFs included four drug-resistance-associated proteins. Two proteins designed as chloramphenicol acetyltransferases (CATs) were encoded by two non-adjacent ORFs, and the other two were TetX2 and a multi-drug ABC transporter permease/ATPase. The putative CAT protein had 62.9 to 79.5% homology to a known type B CAT. The pRA0511 plasmid is the first identified drug-resistance plasmid in R. anatipestifer, more specifically associated with chloramphenicol resistance.
[show abstract][hide abstract] ABSTRACT: Aislamiento y caracterización de un virus de la influenza H5N2 potencialmente patógeno aislado de un pollo en Taiwán en el año 2008. Durante el muestreo de vigilancia para la influenza aviar, se aisló un virus de influenza H5N2 de una muestra de hisopo cloacal de un pollo aparentemente sano en Taiwán en octubre del 2008. Se encontró que la hemaglutinina del virus tenía un par de residuos de aminoácidos dibásicos en el sitio de disociación que podría ser un marcador de alta patogenicidad. Sin embargo, el índice de patogenicidad intravenosa de la cepa fue de 0.89, lo que indica que el virus presentaba tendencia a patogenicidad alta en pollos. El aislamiento del virus fue negativo en 2,916 aves de 146 granjas en un radio de tres kilómetros alrededor de la granja donde se aisló el virus. El análisis genético de los ocho segmentos de la cepa indicaron que el virus aislado era un virus reacomodado genéticamente cuyos segmentos de los genes HA y de la neuraminidasa pertenecían al linaje americano y los genes internos al linaje euroasiático. Abbreviations: AI = avian influenza; AIV = avian influenza virus; HI = hemagglutination-inhibition; HPAIV = high pathogenic avian influenza virus; IVPI = intravenous pathogenicity index; NI = neuraminidase-inhibition; OIE = World Organisation of Animal Health; RT = reverse transcription; SPF = specific pathogen free
[show abstract][hide abstract] ABSTRACT: During the surveillance of avian influenza, an H5N2 influenza A virus was isolated from a cloacal swab sample of an apparently healthy chicken in Taiwan in October 2008. It was found that the HA of the virus had a pair of dibasic amino acid residues at the cleavage site, which might be a marker of highly pathogenic avian influenza virus. However, the intravenous pathogenicity index of the isolate was 0.89, indicating that the virus was approaching high pathogenicity in chickens. Virus isolation was negative in 2916 birds from 146 farms in a 3-km radius around the farm where the virus was isolated. Genetic analysis of the eight segments of the isolate indicated that the isolated virus was a reassortant whose HA and NA gene segments belonged to the American lineage and internal genes to the Eurasian lineage.
[show abstract][hide abstract] ABSTRACT: Canine distemper (CD) is a highly contagious disease with a worldwide distribution. Genetic diversity in genes encoding the haemagglutinin (H) and fusion (F) virus envelope proteins have been implicated in the increasing incidence of CD. Unlike the H gene, little is known about the genetic variability of the F gene in this virus. In the present study sequence analysis of the complete coding region of the F protein from CD virus isolates from Taiwan were carried out. Phylogenetic analysis demonstrated that the majority of isolates were similar to those found in neighbouring China and Japan, but were genetically distinct from vaccine strains. Remarkable variations were found scattered throughout the pre-peptide region (residues 1-135). The sequence identity of this region between locally sourced strains and between these strains and vaccine strains was 89% and 64 to 67%, respectively. Analysis suggested a novel strain of distant genetic lineage was present in dogs in the geographically isolated city of Hualien.
The Veterinary Journal 12/2008; 183(2):184-90. · 2.42 Impact Factor
[show abstract][hide abstract] ABSTRACT: Newcastle disease (ND) and avian influenza (AI) are two of the most important zoonotic viral diseases of birds throughout the world. These two viruses often have a great impact upon the poultry industry. Both viruses are associated with transmission from wild to domestic birds, and often display similar signs that need to be differentiated. A rapid surveillance among wild and domestic birds is important for early disease detection and intervention, and is the basis for what measures should be taken. The surveillance, thus, should be able to differentiate the diseases and provide a detailed analysis of the virus strains. Here, we described a fast, simultaneous and inexpensive approach to the detection of Newcastle disease virus (NDV) and avian influenza virus (AIV) using oligonucleotide microarrays. The NDV pathotypes and the AIV haemagglutinin subtypes H5 and H7 were determined at the same time. Different probes on a microarray targeting the same gene were implemented in order to encompass the diversified virus strains or provide multiple confirmations of the genotype. This ensures good sensitivity and specificity among divergent viruses. Twenty-four virus isolates and twenty-four various combinations of the viruses were tested in this study. All viruses were successfully detected and typed. The hybridization results on microarrays were clearly identified with the naked eyes, with no further imaging equipment needed. The results demonstrate that the detection and typing of multiple viruses can be performed simultaneously and easily using oligonucleotide microarrays. The proposed method may provide potential for rapid surveillance and differential diagnosis of these two important zoonoses in both wild and domestic birds.
[show abstract][hide abstract] ABSTRACT: An outbreak of contagious ecthyma in goats in central Taiwan was investigated. The disease was diagnosed by physical and histopathologic examinations, and the etiology of the disease was identified as orf virus by electron microscopy and polymerase chain reaction (PCR) and sequence of major envelope protein (B2L) gene. The entire protein-coding region of B2L gene were cloned and sequenced. Phylogenetic analysis of B2L amino acid sequences showed that the orf virus identified in this outbreak was closer to the Indian ORFV-Mukteswar 59/05 isolate. This is the first report on the molecular characterization of orf virus in Taiwan.
[show abstract][hide abstract] ABSTRACT: It has been shown that certain slow neurological diseases such as bovine spongiform encephalopathy (also known as "mad cow" disease) could be transmitted through contaminated food intake by animals; therefore, the examination of meat components in commercial feeds is important for the control of the disease in public health. The combination of polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLPs) technique was applied to examine the meat components in dog and cat commercial feeds. The partial nucleotide sequence (359 bp) of animal mitochondrial cytochrome b (cytb, CYT) gene was amplified by PCR and then digested with restriction enzyme Alu I or Mbo I. In this work, eight brands of commercial dog and cat feeds available in Taiwan were examined. All brands of dog feeds that were tested contained meat from four different animals (cattle, pig, goat and chicken). In cat feeds, the chicken meat was found in five out of eight brands.
Journal of Veterinary Medical Science 08/2004; 66(7):855-9. · 0.88 Impact Factor