Hsiang-Jung Tsai

Salmonellae and campylobacters in household and stray dogs in northern Taiwan.

Rectal swabs were collected from 437 household and 491 stray dogs in northern Taiwan from May 2003 to June 2005 to investigate the prevalence and antimicrobial susceptibilities of salmonellae and campylobacters. The results revealed that 2.1% of household dogs and 6.3% of stray dogs were positive for salmonellae, with Salmonella Duesseldorf being the most dominant serotype in both. Additionally, 2.7% of the household dogs and 23.8% of the stray dogs were positive for campylobacters. Campylobacter jejuni was the most prevalent species (86.8%), followed by C. upsaliensis (9.3%) and C. coli (3.9%). Both salmonella and campylobacter isolation rates from the stray dogs were significantly higher than those from the household dogs (p < 0.01). The susceptibility of 33 C. jejuni isolates to eight antimicrobials was studied by the E-test. A high rate of resistance was observed to azithromycin (93.9%), clindamycin (87.9%), erythromycin (81.8%), tetracycline (78.8%), chloramphenicol (69.7%), nalidixic acid (51.5%), gentamicin (33.3%), and ciprofloxacin (18.2%). The susceptibility of 40 Salmonella isolates to 15 antimicrobials was also studied by the disc-diffusion method. All the Salmonella isolates were susceptible to ciprofloxacin and ceftriaxone. Resistance was observed most frequently to tetracycline (77.5%), chloramphenicol (52.5%), and ampicillin (50%).

Prevalence and molecular characterization of chloramphenicol resistance in Riemerella anatipestifer isolated from ducks and geese in Taiwan

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.

Avian hepatitis E virus in chickens, Taiwan, 2013.

A previously unidentified strain of avian hepatitis E virus (aHEV) is now endemic among chickens in Taiwan. Analysis showed that the virus is 81.5%–86.5% similar to other aHEVs. In Taiwan, aHEV infection has been reported in chickens without aHEV exposure, suggesting transmission from asymptomatic cases or repeated introduction through an unknown common source(s).

Detection and Sequence Analysis of Avian Polyomavirus and Psittacine Beak and Feather Disease Virus from Psittacine Birds in Taiwan

Abstract Avian polyomavirus (APV) and psittacine beak and feather disease virus (PBFDV) are the most common viral diseases of psittacine birds. In Taiwan, however, the existence of these viruses in psittacine birds has not been established. Polymerase chain reaction (PCR) methodology was therefore employed to ascertain whether APV and PBFDV genomes were present in isolates from psittacine birds of Taiwan. A total of 165 psittacine birds belonging to 22 genera were examined between 2002 and 2005. Findings revealed an APV-positive rate of 15.2%, a PBFDV-positive rate of 41.2%, and an APV/PBFDV dual infection rate of 10.3%. After cloning and sequencing, sequences of the PCR products were compared with sequences obtained from GenBank. For APV, the nucleotide identity among VP1 and t/T antigen coding regions ranged from 97.5% to 100% and 97.6% to 100%, respectively. For PBFDV, the nucleotide identity of ORF V1 and ORF C1 sequences ranged from 92.2% to 100% and 83.3% to 100%, respectively. The derived amino acid sequence alignment for PBFDV ORF V1 fragments revealed the conservation of two replication motifs and of the nucleotide binding site motif. In PBFDV, six of 42 deduced positions in the ORF C1 amino acid sequence were considered hypervariable. The established phylogenetic trees based on the four genome fragments examined in this study did not allow the assignment of particular APV or PBFDV nucleotide sequences to distinct avian species.

Detection of florfenicol resistance genes in Riemerella anatipestifer isolated from ducks and geese

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.

Infectious Bill Atrophy Syndrome Caused by Parvovirus in a Co-outbreak with Duck Viral Hepatitis in Ducklings in Taiwan

In October 1989, an epizootic duckling disease with high mortality occurred in Taiwan. The disease was characterized by droopiness, inappetence, ataxia, ruffled feathers, and watery diarrhea. Affected ducklings were lame, were unable to stand, showed opisthotonos, and often died 3 or 4 days after the onset of the disease. Tolerant maturing ducklings displayed atrophic upper bills with a protruding tongue and became stunted as they reached maturity. No diagnostic histopathologic lesions were found in these ducklings. Fourteen parvovirus isolates, 33 duck viral hepatitis virus (DVHV) isolates, two adenovirus isolates, and two reovirus isolates were obtained and identified from more than 500 sick ducklings in the epizootic. The epizootic was diagnosed as a co-outbreak of duck parvovirus infections and duck viral hepatitis. The high mortality in ducklings and the bill atrophy syndrome were reproduced in ducklings by inoculating the parvovirus isolates alone. The epizootic was controlled by an emergency immunization program of ducklings with sera collected from recovered ducks or a bivalent inactivated vaccine composed of local DVHV and parvovirus isolates.

Phenotypic and Molecular Characterization of Isolates of Ornithobacterium rhinotracheale from Chickens and Pigeons in Taiwan

Abstract Forty Ornithobacterium rhinotracheale (ORT) strains were isolated from 28 chickens and 12 pigeons for the first time in Taiwan. All isolates reacted positively in the p-nitrophenyl-β-d-galactopyranoside (PNPG) and oxidase tests, showing an API 20NE identification system biocode 0-0-2-0-0-0-4. All the pigeon isolates and 85.7% (24 of 28) of the chicken isolates belonged to serotype A. Compared to the ORT ATCC 51464 strain, 14.3% (4 of 28) of chicken isolates and 58.3% (7 of 12) of pigeon isolates showed smaller colonies after 72 hr incubation. Most of the chicken isolates (22 of 28), but none of the pigeon isolates, could agglutinate chicken and pigeon red blood cells. There appears to be a correlation that ORT isolates with a larger colony size tend to be more able to agglutinate red blood cells than the ORT isolates with a smaller colony size. A majority of isolates was sensitive to amoxicillin, ampicillin, ceftiofur, penicillin, and oxytetracycline. The 16S ribosomal RNA (rRNA) sequences of 23 Taiwanese ORT isolates showed high identity (98%–100%) to sequences in GenBank. Phylogenetic analysis of these sequences showed that pigeon isolates formed a distinctive cluster, while chicken isolates and all other 16S rRNA sequences obtained from GenBank belonged to another two clusters. The results indicate that pigeon ORT isolates are different from most chicken isolates in regard to a number of phenotypic and molecular traits.

Genetic variation of the ompA and 16S rRNA genes of Riemerella anatipestifer.

The genetic diversity of the 16S rRNA and ompA genes of Riemerella anatipestifer was investigated. A 16S rRNA gene-based PCR was able to amplify all 18 Taiwanese strains and 10 reference strains. The identity of 16S rRNA sequence of these strains and seven other sequences retrieved from GenBank was 95.0–100.0%. The percentage identity of the ompA sequence of the 15 Taiwanese strains and eight reference strains amplified in this study and two other sequences retrieved from GenBank was 88.1–100.0%. Phylogenetic analysis based on the 16S rRNA gene showed that all the R. anatipestifer strains fell into a single cluster. It is concluded that the 16S rRNA gene-based PCR is suitable for the screening of R. anatipestifer infections. Phylogenetic analysis of the ompA of R. anatipestifer resulted in three different clusters, while seven clusters were found when the derived amino acid sequence was the basis of analysis. No apparent cluster was found using the criteria of host, isolate serotype, the year or location of isolation.

Swollen head syndrome in Taiwan—isolation of an avian pneumovirus and serological survey

Outbreaks of swollen head syndrome (SHS) were observed in two broiler and two broiler-breeder farms in Taiwan. The disease was characterized by oedematous swelling of the head, especially surrounding the eyelids, the neck and wattles. Avian pneumovirus and Escherichia coli were isolated from birds in all four farms. In addition, Staphylococcus aureus and infectious bursal disease virus were each isolated from one farm. A serological survey of 398 birds from 11 broiler breeder farms showed 86.4% (344) of them had ELISA antibodies against turkey rhinotracheitis virus.

Genetic Variation of Viral Protein 1 Genes of Field Strains of Waterfowl Parvoviruses and Their Attenuated Derivatives

Abstract To understand the genetic variations between the field strains of waterfowl parvoviruses and their attenuated derivatives, we analyzed the complete nucleotide sequences of the viral protein 1 (VP1) genes of nine field strains and two vaccine strains of waterfowl parvoviruses. Sequence comparison of the VP1 proteins showed that these viruses could be divided into goose parvovirus (GPV) related and Muscovy duck parvovirus (MDPV) related groups. The amino acid difference between GPV- and MDPV-related groups ranged from 13.1% to 15.8%, and the most variable region resided in the N terminus of VP2. The vaccine strains of GPV and MDPV exhibited only 1.2% and 0.3% difference in amino acid when compared with their parental field strains, and most of these differences resided in residues 497-575 of VP1, suggesting that these residues might be important for the attenuation of GPV and MDPV. When the GPV strains isolated in 1982 (the strain 82-0308) and in 2001 (the strain 01-1001) were compared, only 0.3% difference in amino acid was found, while MDPV strains isolated in 1990 (the strain 90-0219) and 1997 (the strain 97-0104) showed only 0.4% difference in amino acid. The result indicates that the genome of waterfowl parvovirus had remained highly stable in the field.