Publications by authors named "Takehisa Chuma"

26 Publications

  • Page 1 of 1

Surveillance of Shiga toxin-producing Escherichia coli and Campylobacter spp. in wild Japanese deer (Cervus nippon) and boar (Sus scrofa).

J Vet Med Sci 2020 Sep 13;82(9):1287-1294. Epub 2020 Jul 13.

Laboratory of Veterinary Public Health, Department of Veterinary Medicine, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima, Kagoshima 890-0065, Japan.

Increasing game meat consumption in Japan requires the dissemination of safety information regarding the presence of human pathogens in game animals. Health information regarding the suitability of these animals as a meat source is not widely available. In this study, we aimed to evaluate the safety of game meat and detect potential human pathogens in wild deer (Cervus nippon) and boar (Sus scrofa) in Japan. Fecal samples from 305 wild deer and 248 boars of Yamaguchi, Kagoshima, and Tochigi prefectures collected monthly for 2 years were examined for the prevalence of Shiga toxin-producing Escherichia coli (STEC) and Campylobacter spp. STEC was isolated from 51 deer consistently throughout the year and from three boars; O-antigen genotype O146, the expression of stx2b, and eaeA absence (n=33) were the major characteristics of our STEC isolates. Other serotypes included the medically important O157, stx2b or stx2c, and eaeA-positive (n=4) and O26, stx1a, and eaeA-positive strains (n=1). Campylobacter spp. were isolated from 17 deer and 31 boars. Campylobacter hyointestinalis was the most common species isolated from 17 deer and 25 boars, whereas Campylobacter lanienae and Campylobacter coli were isolated from three and two boars, respectively. Seasonal trends for the isolation of these bacteria were not significant. This study demonstrates that wild game animals carry human pathogens; therefore, detailed knowledge of the safe handling of game meat is needed to prevent foodborne infections.
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http://dx.doi.org/10.1292/jvms.19-0265DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7538328PMC
September 2020

16S rRNA Gene Amplicon Sequence Data from Feces of Five Species of Wild Animals in Japan.

Microbiol Resour Announc 2020 May 28;9(22). Epub 2020 May 28.

Research Center for Food Safety, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan

We report 16S rRNA amplicon sequence data from feces from 58 wild boars, 60 feral raccoons, 9 wild Japanese badgers, 21 wild masked palm civets, and 8 wild raccoon dogs in Japan. The predominant bacterial taxa in the fecal microbiota were similar in part but varied among the animal species.
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http://dx.doi.org/10.1128/MRA.00368-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7256260PMC
May 2020

16S rRNA Gene Amplicon Sequence Data from Feces of Wild Deer (Cervus nippon) in Japan.

Microbiol Resour Announc 2020 May 28;9(22). Epub 2020 May 28.

Research Center for Food Safety, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan

We report 16S rRNA amplicon sequence data from feces of 109 wild deer in Japan. The dominant bacterial taxa in fecal microbiota of wild deer hunted between village and mountainous areas and those living on Miyajima Island and in Nara Park were similar but differed in abundance.
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http://dx.doi.org/10.1128/MRA.00346-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7256258PMC
May 2020

α-Sens: The improved ARE-Nrf2-based sensitization screening assay using normalized transcriptional activity.

Toxicology 2020 06 23;439:152476. Epub 2020 Apr 23.

The United Graduate School of Veterinary Science, Yamaguchi University, Japan.

Two non-animal test methods, KeratinoSens™ and LuSens, have been approved by the Organization of Economic Cooperation and Development (OECD) test guidelines for evaluating the sensitization potential of chemicals, and been positioned as a method for appraising key event (KE)-2, namely, the keratinocyte response component of the Adverse Outcome Pathway (AOP) in sensitization process. However, these two methods require separate cytotoxicity tests to determine the concentrations to be tested in the main test. Therefore, we developed a simple and highly accurate KE-2 test method named α-Sens that uses the dual luciferase assay system and attempted a further application of luciferase-based determination of cell viability to calculate the normalized Antioxidant response element (ARE)-mediated transcriptional activity, named normalized ARE Activity (nAA), to evaluate the sensitizing potential of chemicals. A cell line carrying the ARE-inducible Firefly luciferase reporter gene and Thymidine kinase (TK) promoter-driven Renilla luciferase gene was established and used for the α-Sens. A total of 28 chemicals, consisting of 19 skin sensitizers and nine non-skin sensitizers were tested by this assay system. The α-Sens yielded an accuracy (%), sensitivity (%), and specificity (%) against corresponding values for local lymph node assay of 96.4 %, 95.0 %, and 100 %, respectively, and for human data of 100 % for all. The α-Sens gave clear positive results for phenyl benzoate and eugenol, chemicals for which KeratinoSens™ or LuSens yielded false-negative results, using a new parameter. Our results suggest that better prediction capacity could be achieved by using nAA as a classifier compared to other existing KE-2 test methods. In conclusion, the α-Sens is promising as a simple and highly accurate in vitro skin sensitization test method for evaluation of KE-2.
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http://dx.doi.org/10.1016/j.tox.2020.152476DOI Listing
June 2020

Increased Salmonella Schwarzengrund prevalence and antimicrobial susceptibility of Salmonella enterica isolated from broiler chickens in Kagoshima Prefecture in Japan between 2013 and 2016.

J Vet Med Sci 2020 May 25;82(5):585-589. Epub 2020 Mar 25.

Laboratory of Veterinary Public Health, Department of Veterinary Medicine, Joint Faculty of Veterinary Medicine, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan.

This study aimed to analyze the Salmonella serovars, measure the minimum inhibitory concentration of antimicrobials, and examine the antimicrobial resistance genes of Salmonella isolated from 192 broiler flocks in Kagoshima Prefecture in Japan, from 2013 to 2016. We found that all Salmonella isolates belonged to three serovars: Salmonella Manhattan, S. Infantis, and S. Schwarzengrund. Among them, S. Schwarzengrund prevalence has recently increased annually making the main serovar. Most recovered isolates were highly resistant to streptomycin, sulfamethoxazole, and oxytetracycline. We saw the reduction of third-generation cephalosporin resistance and identified the reason of increased kanamycin resistance to be the increased number of S. Schwazengrund isolates. Among the kanamycin-resistant Salmonella isolates, aphA1 constituted the main resistance gene detected.
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http://dx.doi.org/10.1292/jvms.20-0096DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7273603PMC
May 2020

Differences in phenotypic and genetic characteristics of Trueperella pyogenes detected in slaughtered cattle and pigs with septicemia.

J Vet Med Sci 2020 May 24;82(5):626-631. Epub 2020 Mar 24.

Laboratory of Veterinary Public Health, Joint Faculty of Veterinary Medicine, Kagoshima University, Korimoto 1-21-24, Kagoshima-shi, Kagoshima 890-0065, Japan.

We investigated the hemolytic properties, biochemical properties, and possession of virulence factor genes of Trueperella pyogenes isolated from cattle and pigs with septicemia. The porcine strains showed significantly stronger hemolyticity than the bovine strains. In addition, T. pyogenes from cattle and pigs also differed in biochemical properties. Virulence factor genes (nanP, cbpA, fimC, and fimE) were more prevalent in bovine strains, whereas other virulence factor genes (nanH and fimG) were more prevalent in porcine strains. T. pyogenes isolated from pig and cattle with septis cases in Japanese meat inspection showed variability in biochemical and genetic properties. Differences were observed between porcine and bovine strain in term of the hemolytic strength and possession of genes for factors promoting adhesions which are considered pathogenic.
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http://dx.doi.org/10.1292/jvms.19-0370DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7273585PMC
May 2020

Surveillance of antibiotic resistance in Escherichia coli isolated from wild cranes on the Izumi plain in Kagoshima prefecture, Japan.

J Vet Med Sci 2019 Sep 11;81(9):1291-1293. Epub 2019 Jul 11.

Joint Graduate School of Veterinary Medicine, Kagoshima University, 1-21-24 Kagoshima-shi, Kagoshima 890-0065, Japan.

The prevalence of antibiotic resistance in 376 Escherichia coli (E. coli) isolates from fecal samples of Hooded and White-naped cranes was investigated on the Izumi plain in Kagoshima prefecture, Japan, during winter 2016 and 2017. Resistance to oxytetracycline, ampicillin, and nalidixic acid were observed in 10.9%, 3.1-4.4%, and 2.1-7.7% of isolates, respectively. Since the previous surveillance in 2007, isolation rates of antibiotic-resistant E. coli recovered from wild cranes have remained at significantly low levels compared with those in Japanese livestock. Our results indicate that surveillance of antibiotic-resistant E. coli from wild cranes wintering in the Izumi plain could be a useful strategy to indicate natural environmental pollution by antibiotic-resistant bacteria in the environment.
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http://dx.doi.org/10.1292/jvms.19-0305DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6785602PMC
September 2019

Prevalence of Salmonella in broiler chickens in Kagoshima, Japan in 2009 to 2012 and the relationship between serovars changing and antimicrobial resistance.

BMC Vet Res 2019 Apr 8;15(1):108. Epub 2019 Apr 8.

Laboratory of Veterinary Public Health, Joint Faculty of Veterinary Medicine, Kagoshima University, 1-21-24 Korimoto, Kagoshima, 890-0065, Japan.

Background: This study aimed to examine the prevalence, serovars, and antimicrobial resistance of Salmonella isolates from broiler chickens in Kagoshima, Japan. A total of 192 flocks and 3071 samples were collected from broiler chickens at local farms in Kagoshima, Japan from 2009 to 2012.

Result: Among the tested farms, 49.0% of flocks were positive for Salmonella, and 243 isolates were obtained from 3071 cecal samples (7.9%). All the Salmonella isolates were one of three serovars: S. Infantis (57.6%); (140/243), S. Manhattan (40.3%; 98/243 and S. Schwarzengrund (2.1%; 5/243). The proportion of S. Infantis isolates decreased from 66.0% in 2009 to 50.0% in 2011 but increased to 57.6% in 2012, while the proportion of S. Manhattan isolates significantly increased from 26.4 to 50% from 2009 to 2011, and decreased moderately to 40.9% in 2012. Most of the recovered Salmonella isolates were resistant to three antimicrobials, i.e., streptomycin (95.1%), sulfamethoxazole (91.0%) and oxytetracycline (91.4%). In contrast, all Salmonella strains were susceptible to chloramphenicol. Comparison of this study to previous studies of the antimicrobial susceptibility of Salmonella isolates showed that: the percentage of antibiotic-resistance isolates increased dramatically for two antibiotics, ampicillin (from 22.4 to 55.1%) and cefotaxime (from 9.1 to 52.7%). In contrast, the percentage of ofloxacin-resistant isolates decreased across the three survey periods, from 20.8% in 2004-2006 to 1.6% in the present study period (2009-2012). In addition, S. Infantis exhibited a variety of resistance to antimicrobials examined from sensitive to resistance to eight antimicrobials. Multidrug resistance to more than 6 six antimicrobials was detected in 113 (46.5%) of the isolates, and most of them were S. Manhattan.

Conclusions: There was a marked change in the serovars and antimicrobial resistance profiles of the Salmonella isolates in this study compared to those in previous studies. The percentage of S. Manhattan isolates increased as did the percentages of ampicillin- and cefotaxime-resistant isolates.
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http://dx.doi.org/10.1186/s12917-019-1836-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6454760PMC
April 2019

Chronological Change of Resistance to β-Lactams in Salmonella enterica serovar Infantis Isolated from Broilers in Japan.

Front Microbiol 2013 21;4:113. Epub 2013 May 21.

Laboratory of Veterinary Public Health, Department of Veterinary Medicine, Joint Faculty of Veterinary Medicine, Kagoshima University Kagoshima, Japan.

Epidemiologic surveillance study was conducted in southern Japan to determine the antimicrobial resistance phenotypes and characterize the β-lactamase genes and the plasmids harboring these genes in Salmonella enterica serovar Infantis (S. Infantis) isolates from broilers. Between January, 2007 and December, 2008, a total of 1,472 fecal samples were collected and examined at the Laboratory of Veterinary Public Health, Kagoshima University, Japan. In 93 (6.3%) isolates recovered, 33 (35.5%) isolates showed resistance to cefotaxime, an extended-spectrum cephalosporin (ESC), conferred by TEM-20, TEM-52 and CTX-M-25 extended-spectrum β-lactamases (ESBLs). In addition to ESC-resistance, eight (8.6%) isolates exhibited resistance to cefoxitin mediated by CMY-2 AmpC β-lactamase. Plasmid analysis and polymerase chain reaction replicon typing revealed the bla TEM-20 and bla CMY-2 genes were associated with IncP plasmids, bla TEM-52 was linked with a non-typable plasmid and bla CTX-M-25 was carried by an IncA/C plasmid. Non-β-lactam resistance to streptomycin, sulfamethoxazole, and oxytetracycline encoded by the aadA1, sul1, and tet(A) genes, respectively, was found in 86 (92.5%) isolates. Resistance to kanamycin and ofloxacin was exhibited in 12 (12.9%) and 11 (11.8%) isolates, respectively, the former was mediated by aphA1-Iab. These data indicate that S. Infantis isolates producing ESBLs and AmpC β-lactamase have spread among broiler farms in Japan. These data demonstrated that the incidence of ESC-resistant S. Infantis carrying bla TEM-52 remarkably increased and S. Infantis strains harboring bla CMY-2, bla TEM-20, or bla CTX-M-25 genes emerged from broilers in Japan for the first time in 2007 and 2008.
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http://dx.doi.org/10.3389/fmicb.2013.00113DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3659313PMC
June 2013

Prevalence and epidemiological relationship of CMY-2 AmpC β-lactamase and CTX-M extended-spectrum β-lactamase-producing Escherichia coli isolates from broiler farms in Japan.

J Vet Med Sci 2013 15;75(8):1009-15. Epub 2013 Mar 15.

Yamaguchi Prefectural Institute of Public Health and Environment, 2-5-67 Aoi, Yamaguchi 753-0821, Japan.

To evaluate the prevalence of extended-spectrum cephalosporin (ESC)-resistant Enterobacteriaceae in broiler chickens, 41 rectal samples taken from 4 commercial farms were examined. Desoxycholate hydrogen sulfide lactose agars, supplemented with either 4 μg/ml cefotaxime or 16 μg/ml ceftazidime, were used to screen ESC-resistant bacteria. ESC-resistant bacteria were isolated from all samples. Of the 164 ESC-resistant bacteria (included 4 isolates per a sample), 163 were Escherichia coli, while 1 isolate was identified as Enterobacter cloacae. Extended-spectrum β-lactamase (ESBL) genes and plasmid-mediated AmpC β-lactamase genes in the isolates were determined by PCR and sequencing. One AmpC β-lactamase gene, bla(CMY-2) (66%), and 4 ESBL genes, bla(CTX-M-1) (26%), bla(CTX-M-55) (10%), bla(SHV-5) (4%) and bla(CTX-M-2) (3%), were detected in the E. coli isolates. The epidemiological relationship of the CMY-2 and CTX-M β-lactamase-producing isolates among the farms was analyzed by pulsed-field gel electrophoresis using the XbaI restriction enzyme. Forty-one (Y1-Y41) and 14 (X1-X14) clusters were found in the CMY-2 and CTX-M-carrying E. coli isolates, respectively. Some clusters included isolates derived from more than 1 farm, indicating some cross-contamination of clonal strains and spread of CMY-2 AmpC β-lactamase or CTX-M ESBL among the farms.
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http://dx.doi.org/10.1292/jvms.12-0453DOI Listing
April 2014

Emergence of Salmonella enterica serovar infantis harboring IncI1 plasmid with bla(CTX-M-14) in a broiler farm in Japan.

J Vet Med Sci 2012 Sep 16;74(9):1213-6. Epub 2012 May 16.

Yamaguchi Prefectural Institute of Public Health and Environment, 2-5-67 Aoi, Yamaguchi 753-0821, Japan.

Cefotaxime (CTX)-resistant and -susceptible Salmonella enterica serovar Infantis isolates obtained from broilers raised on a farm in January 2010 in Japan were characterized to establish their resistance determinants. The CTX-resistant isolates produced CTX-M-14 extended-spectrum β-lactamase and harbored 2 distinct plasmid of approximately 140- and 95-kb, whereas the CTX-susceptible isolates harbored one 140-kb plasmid. The 95-kb plasmids were replicon typed as IncI1 carrying the bla(CTX-M-14) gene, while the 140-kb plasmids were IncP and harbored the aphA1, aadA1, tetA, and sul1 genes. Genetic fingerprinting by pulsed-field gel electrophoresis revealed similar macrorestriction profiles amongst CTX-resistant and susceptible isolates, suggesting a clonal relationship. The presence of CTX-resistant S. Infantis on a broiler farm has occurred through the acquisition of IncI1 resistance plasmid.
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http://dx.doi.org/10.1292/jvms.11-0488DOI Listing
September 2012

Effect of cooled and chlorinated chiller water on Campylobacter and coliform counts on broiler carcasses during chilling at a middle-size poultry processing plant.

J Vet Med Sci 2012 Jan 6;74(1):129-33. Epub 2011 Sep 6.

Iwakuni Health Welfare and Center of Yamaguchi Prefecture, Japan.

To evaluate the effect of cooled and chlorinated chill water for Campylobacter and coliforms at a middle-size processing plant which was considered to be difficult for eliminate pathogenic bacteria on carcasses, following three conditions were examined; keeping temperature at < 20, < 10 and < 10°C, and chlorine concentration at < 50, < 50 and 50 to 70 ppm during processing in experiment 1, 2 and 3 respectively. Fifteen prechill and 15 postchill carcasses were examined in each experiment. In lower temperature of experiment 2, decreasing rate (%) of coliforms was significantly higher (P<0.01) than that in experiment 1. In higher chlorination of experiment 3, no Campylobacter was detected from all postchill carcasses.
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http://dx.doi.org/10.1292/jvms.11-0167DOI Listing
January 2012

Re-emergence of multi-drug resistant Salmonella enterica serovar Stanley from cattle.

Vet Res Commun 2011 Jan;35(1):55-60

Laboratory of Veterinary Public Health, Department of Veterinary Medicine, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan.

During 2009, Salmonella enterica subspecies enterica serovar Stanley isolates were recovered from cattle diagnostic specimens in southern Japan, and the isolates were examined to characterize the genetic determinants involved in this new pathogenicity that associated with mortality in cattle. All the isolates were multi-drug resistance exhibited resistance to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, oxytetracycline, and kanamycin (ACSSuT-Km) encoded by blaTEM, catA, aadA1, sul1, tet(A), and aphA1 genes, respectively. Class 1 integrons of 1.5-kb size were detected in all MDR isolates. The isolates harboured easily transferable plasmids of ca. 210-kb with the potential of transmitting resistance phenotype and genotype detected in the donor isolates. XbaI-digested PFGE patterns generated two related clusters implicated in the dissemination of multi-drug resistance amongst Salmonella Stanley isolates. An emergence of multi-drug resistant Salmonella Stanley amongst food-producing animals, including cattle is a threat to human health, as resistant isolates may be transmitted to humans through the food chain.
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http://dx.doi.org/10.1007/s11259-010-9448-4DOI Listing
January 2011

Characterization of antibiotic resistance and the emergence of AmpC-producing Salmonella Infantis from pigs.

J Vet Med Sci 2010 Nov 7;72(11):1437-42. Epub 2010 Jul 7.

Laboratory of Veterinary Public Health, Department of Veterinary Medicine, Faculty of Agriculture, Kagoshima University, Kagoshima, Japan.

During the period of 2007-2008, a total of 270 pig fecal samples were collected from a meat processing plant located in southern Japan and examined for Salmonella species. A total of 44 Salmonella isolates were recovered, and antimicrobial resistance was detected in serotypes Typhimurium (n=9), Infantis and Choleraesuis (n=2), and Derby, Miyazaki and Schwarzengrund (n=1). Multidrug resistance was seen in serotypes Typhimurium (n=8) and Infantis (n=2). The most commonly observed resistance phenotypes were against streptomycin, oxytetracycline and sulfamethoxazole (100%), ampicillin (90%), chloramphenicol (50%), cephalothin (30%) and cefoxitin, ceftazidime and kanamycin (each 20%). Extended-spectrum cephalosporin-resistant Salmonella Infantis isolates producing plasmid-mediated, bla(CMY-2) gene were detected. These AmpC-producing isolates showed resistance to ampicillin and cephems (cephalothin, cefoxitin and ceftazidime). Resistance transfer experiments showed that transconjugants and transformants coexpressed resistance phenotypes similar to the donor isolates. To the best of our knowledge, this is the first report worldwide describing serovar Infantis from pigs capable of producing AmpC β-lactamase. Then, we detected the pentadrug-resistance phenotype in Salmonella Typhimurium isolates, which yielded class 1 integron amplicons of 1.0 and 1.2 kb. Genetic fingerprinting analysis by pulsed-field gel electrophoresis and an assay by polymerase chain reaction confirmed the isolates to be Salmonella Typhimurium DT104. In conclusion, the findings of this survey call for the systematic and comprehensive domestic and international surveillance programs to determine the true rates of occurrence of AmpC-producing Salmonella both in the livestock and public health sectors.
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http://dx.doi.org/10.1292/jvms.10-0186DOI Listing
November 2010

Genetic analysis of multidrug-resistant Salmonella enterica serovars Stanley and Typhimurium from cattle.

Vet Microbiol 2010 Sep 3;145(1-2):76-83. Epub 2010 Mar 3.

Laboratory of Veterinary Public Health, Department of Veterinary Medicine, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan.

During 2005-2008, a longitudinal study was conducted in southern Japan to detect and characterize multidrug-resistant Salmonella enterica serovars recovered from cattle diagnostic specimens. Determination of antimicrobial resistance phenotypes and genotypes, identification of Salmonella genomic island 1 (SGI1), detection of virulence genes, plasmid analysis, conjugal transfer experiments, and sequencing of class 1 integrons were conducted. Multidrug-resistant Salmonella detected were serovars Stanley, Typhimurium, and O4:d. Salmonella Stanley isolates exhibited resistance to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, oxytetracycline, trimethoprim, and kanamycin (ACSSuT+) encoded by bla(TEM), catA, aadA2, tetA, sul1, dfrA12, and aphA1 genes, respectively. Sequencing analysis revealed that aadA2 and dfrA12 were integrated as gene cassettes within the class 1 integrons of 1.5kb size. Importantly, the isolates harboured easily transferable plasmids of ca. 210kb with the potential of transmitting resistance phenotype and genotype detected in the donor isolates. Moreover, Salmonella Typhimurium DT104 isolates with typical SGI1 were detected and presented ACSSuT+ resistance phenotype encoded by bla(PSE-1) and bla(TEM); floR; aadA1; sul1; and tetA and tetG, respectively. Salmonella Typhimurium isolates carried plasmids of variable sizes ranging from 3.5 to 100 kb with DT104 isolates harbouring plasmids of ca. 90 kb. Salmonella serovar O4:d had ACSSuT+ resistance phenotype mediated by bla(TEM), catA, aadA1, sul1, tetA, and aphA1 genes. A virulence gene invA was found in all multidrug-resistant Salmonella Typhimurium, Stanley and O4:d clinical isolates. In conclusion, this is the first report describing the occurrence of multidrug-resistant Salmonella Stanley from bovine species. The emergence of Salmonella Stanley isolates exhibiting plasmid-encoded high-level multidrug resistance is an important health concern because this new pathogenecity was associated with mortality in cattle.
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http://dx.doi.org/10.1016/j.vetmic.2010.02.035DOI Listing
September 2010

Antimicrobial susceptibility of Erysipelothrix rhusiopathiae isolated from pigs in Southern Japan with a modified agar dilution method.

J Vet Med Sci 2010 May 13;72(5):643-5. Epub 2010 Jan 13.

Laboratory of Veterinary Public Health, Department of Veterinary Medicine, Faculty of Agriculture, Kagoshima University, Korimoto, Kagoshima890-0065, Japan.

The determination of antimicrobial minimum inhibitory concentration (MIC) in Erysipelothrix rhusiopathiae by using the agar dilution method has not been covered by the Clinical and Laboratory Institute (CLSI). Only the broth microdilution method has been outlined. This report describes a modification of the agar dilution procedure for E. rhusiopathiae using Trypto-soy agar supplemented with 0.1% Tween 80 and incubation in ambient air at 37 degrees C for 24 hr. The MICs of the assay were in agreement with those of the broth microdilution method recommended by the CLSI. Antimicrobial susceptibility test was performed using this method for 149 E. rhusiopathiae isolates from 2 meat processing plants in Kagoshima Prefecture during the period of April 2004 to March 2005. The number of strains resistant to oxytetracycline, erythromycin, lincomycin, ofloxacin and enrofloxacin were 56 (37.6%), 4 (2.7%), 18 (12.1%), 21 (14.1%) and 19 (12.8%), respectively. All strains were susceptible to ampicillin.
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http://dx.doi.org/10.1292/jvms.09-0448DOI Listing
May 2010

Detection and characterization of extended-spectrum beta-lactamase (TEM-52)-producing Salmonella serotype Infantis from broilers in Japan.

Foodborne Pathog Dis 2010 May;7(5):515-21

Department of Veterinary Medicine, Faculty of Agriculture, Kagoshima University, Kagoshima, Japan.

During 2004 and 2006, multidrug-resistant Salmonella enterica subspecies enterica serovar Infantis (Salmonella Infantis) isolates (n = 120) were recovered from broiler cecal samples collected from a meat-processing plant, and the isolates were examined. The study was conducted to detect and characterize extended-spectrum beta-lactamase (ESBL)-producing Salmonella Infantis isolates recovered from broiler chickens and determine the mechanisms of transfer of the resistance traits. Extended-spectrum cephalosporins-resistant Salmonella Infantis isolates producing ESBL TEM-52 were detected. The mutant bla(TEM-52) gene and the wild-type bla(TEM-1) gene that mediated resistance to ampicillin (an extended-spectrum penicillin) and cephalothin (a narrow-spectrum cephalosporin) were located on approximately 50-kb conjugative plasmids among beta-lactam-resistant (n = 29) isolates. The bla(TEM) genes did not cotransfer with aadA1, sul1 (both associated with class 1 integrons), tetA, and dfrA5, signifying a chromosomal location of these non-beta-lactam resistance-encoding genes. This is the first report describing TEM-52-producing S. enterica from food-producing animals in Japan. An emergence of TEM-type ESBL is an important concern to public health because this readily transferable resistance mechanism threatens the value of the third-generation cephalosporins and may reduce the clinical utility of this class of antibiotics against pathogenic Gram-negative bacteria.
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http://dx.doi.org/10.1089/fpd.2009.0454DOI Listing
May 2010

Antimicrobial susceptibility of Treponema phagedenis-like spirochetes isolated from dairy cattle with papillomatous digital dermatitis lesions in Japan.

J Vet Med Sci 2010 Mar 8;72(3):379-82. Epub 2009 Dec 8.

Laboratory of Veterinary Public Health, Department of Veterinary Science, Faculty of Agriculture, University of Miyazaki, 1-1 Gakuenkibanadai-nishi, Miyazaki 889-2192, Japan.

The minimal inhibitory concentrations (MICs) of 23 Treponema phagedenis-like spirochetes isolated from dairy cattle with papillomatous digital dermatitis (PDD) lesions in Japan were investigated by a broth microdilution method using 15 antimicrobial agents. Although all MIC values showed a monomodal distribution, the MICs of the antimicrobial agents for 90% (MIC(90)) of the isolates tested varied among the agents examined. The MIC(90) values for penicillin G, ampicillin, and erythromycin were <0.06 microg/ml. In contrast, the MIC(90) values for kanamycin, streptomycin, rifampicin, sulfamethoxazole, trimethoprim, and colistin were >128 microg/ml. Oxytetracycline, lincomycin, enrofloxacin, chloramphenicol, ceftiofur, and gentamicin showed intermediate values, i.e., 0.5~32 microg/ml. The present study suggested that no isolate had acquired resistance to the antimicrobial agents examined, although they may have natural resistance to some agents. Furthermore, the in vitro antimicrobial susceptibility data would provide helpful information for PDD treatment and the development of a selective medium for isolating the organism effectively.
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http://dx.doi.org/10.1292/jvms.09-0418DOI Listing
March 2010

Successful treatment of bacillary hemoglobinuria in Japanese Black cows.

J Vet Med Sci 2009 Aug;71(8):1105-8

Laboratory of Farm Animal Production Medicine, Kagoshima University, Kagoshima, Japan.

Four pasture-fed Japanese Black cows showed the main clinical symptoms of severe hemoglobinuria at different periods between 2003 and 2007. Hematological analyses at the first consultation revealed severe anemia, and biochemical analyses indicated both severe hemolysis and disruption of hepatic function. Although the first 2 patients died, the hemoglobinuria and general condition of the remaining 2 cows, who were immediately initiated on large doses of antibiotics, improved within 3 days. Clostridium haemolyticum was detected by polymerase chain reaction (PCR) analysis of the blood sample of 1 of the infected cows. Anti-fascioliasis medicine is administered, and since then, no case of hemoglobinuria has been observed. The cows were diagnosed with bacillary hemoglobinuria, and they represent the first few cases in Japan.
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http://dx.doi.org/10.1292/jvms.71.1105DOI Listing
August 2009

Genetic analysis of multi-drug resistance and the clonal dissemination of beta-lactam resistance in Salmonella Infantis isolated from broilers.

Vet Microbiol 2010 Jan 10;140(1-2):136-41. Epub 2009 Jul 10.

Laboratory of Veterinary Public Health, Department of Veterinary Medicine, Faculty of Agriculture, Kagoshima University, 1 21-24 Korimoto, Kagoshima 890-0065, Japan.

An epidemiologic study was conducted to investigate the incidence and characterize the antimicrobial resistance determinants, analyzing plasmid profiles, and establishing the genetic relationship among beta-lactam-resistant isolates of Salmonella Infantis from broilers in Southern Japan. A total of 120 isolates were recovered from 56 flocks belonging to 44 holdings during 2004-2006. The percentages of resistance were as follows: ampicillin (24%), cephalothin (23%), cefoxitin (0%), ceftazidime (11%), cefotaxime (11%), chloramphenicol (0%), kanamycin (7.5%), ofloxacin (20%), oxytetracycline, streptomycin and sulfamethoxazole (100%) and trimethoprim (75%). The incidence of bla(TEM)-encoded beta-lactam resistance in 2004-2006 was significantly higher than in 1998-2003 (P<0.001). BlnI-digested PFGE patterns generated two related clusters implicated in the dissemination of beta-lactam resistance. Two types of plasmid profiles were observed and two plasmids of ca. 50 and 180-kb size were carried by beta-lactam-resistant isolates. Streptomycin resistance was conferred by aadA1 (n=116), aadA1-aadA2 (n=1), and aadA1-strA-strB (n=3). Resistances to kanamycin, oxytetracycline, sulfamethoxazole and trimethoprim were conferred by aphA1 (n=9, 100%), tetA (n=120, 100%) sul1 (n=120, 100%) and dfrA5 (n=90, 100%), respectively. Two types of class 1 integrons were detected: 1.0 kb (n=120) and, 1.0/1.5 kb (n=3). Integrons of 1.0/1.5 kb were found in isolates with the aadA1-strA-strB gene combination. For the first time, all S. Infantis isolates showed resistance to at least three classes of antimicrobial agents; and the intestinal tract of healthy poultry was a reservoir of the extended-spectrum cephalosporin-resistant isolates of serovar Infantis.
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http://dx.doi.org/10.1016/j.vetmic.2009.07.007DOI Listing
January 2010

Antimicrobial susceptibility phenotypes, resistance determinants and DNA fingerprints of Salmonella enterica serotype Typhimurium isolated from bovine in Southern Japan.

Int J Antimicrob Agents 2007 Aug 31;30(2):150-6. Epub 2007 May 31.

Laboratory of Veterinary Public Health, Department of Veterinary Medicine, Faculty of Agriculture, Kagoshima University, 1 21-24 Korimoto, Kagoshima 890-0065, Japan.

A longitudinal study was conducted in cattle to determine the antimicrobial resistance phenotypes, integron elements, resistance genes and pulsed-field gel electrophoresis fingerprints among Salmonella enterica serotype Typhimurium isolates. A total of 33 strains were isolated and categorised into Groups A, B and C during the period 1989-2004. Thirty-one strains (93.9%) showed resistance to ampicillin (A) encoded by bla(OXA-1), bla(TEM) and bla(PSE-1) genes; 84.8% showed resistance to chloramphenicol (C) encoded by floR and catA1; 97.0% were resistant both to streptomycin (S) and sulfamethoxazole (Su), the former encoded by aadA1 and aadA2; 100% were resistant to oxytetracycline (T) encoded by tetA, tetB and tetG; and 42.4% were resistant to kanamycin (Km) encoded by aphA1-Iab. Multidrug resistance types observed were ACSSuT-Km (n=13), ACSSuT (n=15), ASSuT (n=3) and SSuT (n=2). Class 1 integrons ranging from 1.0 kb to 1.9 kb were detected from 54.5% of isolates (18/33). Integrons were not detected initially (1989-1992), then during the 1993-1996 interval a high frequency of 1.0 kb and 1.2kb amplicons were detected and during 2000-2004 the amplicon size increased to 1.7 kb and 1.9 kb. We report evidence of additional integration of resistance gene cassettes as shown by integrons with increased size. Finally, group B strains showed banding patterns indistinguishable from S. Typhimurium DT104 reference strain, indicating that the DT104 lineage existed on the island since 1993.
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http://dx.doi.org/10.1016/j.ijantimicag.2007.03.017DOI Listing
August 2007

Absence of correlation between karyotype profiles of Trypanosoma congolense and resistance to isometamidium chloride.

Vet Parasitol 2007 Jul 29;147(3-4):311-4. Epub 2007 May 29.

Institute for Parasitology and Tropical Veterinary Medicine, Freie Universität Berlin, Konigsweg 67, D-14163 Berlin, Germany.

Chromosome profiles of 10 Trypanosoma (T.) congolense populations with different isometamidium sensitivities were compared using the pulsed field gel electrophoresis technique. The aim was to elucidate whether there was a karyotype pattern specific to eight isometamidium resistant phenotypes. Analysis of the profiles indicated that all populations displayed several discrete bands at the region of small, intermediate and large chromosomes. The highest similarity was observed between two isolates originating from Burkina Faso, indicating that they had the same genetic origin. Other eight strains exhibited different patterns in terms of chromosome size and numbers such that there was no characteristic karyotype pattern that was established specifically to identify resistant populations and discriminate them from the sensitive ones. This study has revealed that isometamidium resistance is not correlated to karyotype profile in T. congolense.
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http://dx.doi.org/10.1016/j.vetpar.2007.04.016DOI Listing
July 2007

Genotyping and quantitation of noroviruses in oysters from two distinct sea areas in Japan.

Microbiol Immunol 2007 ;51(2):177-84

Yamaguchi Prefectural Research Institute of Public Health, Yamaguchi, Japan.

Norovirus (NV) is a causative agent of acute gastroenteritis in humans, and shellfishes including oysters act as major vehicles of the virus. To investigate the genetic characteristics of NVs, we collected 1,512 oysters for raw consumption between October 2002 and March 2005 from two distinct areas (area A: the Sanriku Sea area; area B: the Setouchi Sea area). We detected the capsid gene and subjected it to phylogenetic analysis. By further quantification of the copy number of the genome by using real-time PCR, the NV capcid gene was detected in approximately 5% of the oysters, and they showed wide diversity. Two percent of the oysters from area B showed relatively large number of NVs, i.e., over 100 copies of capsid gene/oyster, whereas this was not observed in area A. Most of the detected NVs from oysters and humans were genetically related when the capsid region was compared. These results suggested that NVs obtained from humans and those obtained from oysters showed a potential relationship to each other and that some populations of Japanese oysters accumulated a relatively large number of NVs.
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http://dx.doi.org/10.1111/j.1348-0421.2007.tb03899.xDOI Listing
May 2007

Molecular epidemiology of antimicrobial resistance among Salmonella enterica serovar Infantis from poultry in Kagoshima, Japan.

Int J Antimicrob Agents 2006 Oct 1;28(4):302-7. Epub 2006 Sep 1.

Laboratory of Veterinary Public Health, Department of Veterinary Medicine, Faculty of Agriculture, Kagoshima University, 1 21-24 Korimoto, Kagoshima 890-0065, Japan.

Antimicrobial susceptibility and resistance genes of 135 strains of Salmonella enterica serovar Infantis isolated from poultry in Kagoshima were examined. One strain (0.7%) was resistant to ampicillin (A), 97% to streptomycin (S), 95.6% to sulphamethoxazole (Su), 96.3% to oxytetracycline (T), 11.1% to kanamycin (Km) and 36.3% to ofloxacin (O). Multiresistant phenotypes identified were ASSuT-Km, SSuT-Km, SSuT-O and SSuT. Class 1 integrons were detected in 94.8% of isolates. Approximately 89% of oxytetracycline-resistant strains carried the tetA gene and all of the 131 streptomycin-resistant isolates carried the aadA1a gene. Forty-percent of kanamycin-resistant isolates carried the aphA1-Iab gene. All isolates were susceptible to chloramphenicol. Recognition of TEM-type beta-lactamase in a S. Infantis strain from chickens is a recent rare finding in Japan.
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http://dx.doi.org/10.1016/j.ijantimicag.2006.07.003DOI Listing
October 2006

Analysis of Campylobacter spp. contamination in broilers from the farm to the final meat cuts by using restriction fragment length polymorphism of the polymerase chain reaction products.

Int J Food Microbiol 2006 Aug 27;110(3):240-5. Epub 2006 Jun 27.

Department of Veterinary Public Health, Faculty of Agriculture, Kagoshima University, Kagoshima 890-0065, Japan.

We investigated the genotype diversity and dynamics of Campylobacter jejuni and Campylobacter coli in six commercial broiler farms during rearing and abattoir processing. In total, 223 C. jejuni and 36 C. coli strains isolated (on farm, transportation crates, carcasses after defeathering, and chicken wing meat at the end of the processing line) were subtyped by PCR-RFLP based on flagellin (fla typing) gene. Eleven (C. jejuni) and four (C. coli) different RFLP patterns were found. Multiple C. jejuni genotypes were identified in five out of six farms (except Farm 5). Furthermore, a clear tendency for dominance of particular genotypes was observed in almost all farms except Farm 3. Although diverse C. jejuni genotypes were isolated on the farms and transport crates, they were not detected in chicken wing cuts at the end of the processing line. We also observed varied distribution of types in different sampling stages both at the farm level and the processing environment. However, the interpretation of such fluctuations is precarious as new types occurred on some occasions, particularly during processing. Our results show that chicken wing meat contamination resulted mainly from farm strain carryover, and that the carcasses were probably contaminated during processing. In addition, the new strain types observed were isolated more frequently after defeathering as compared to other processing steps. Therefore, this stage, in addition to evisceration, is one of the critical control points in the processing line to prevent cross-contamination and for controlling the spread of campylobacters.
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http://dx.doi.org/10.1016/j.ijfoodmicro.2006.04.043DOI Listing
August 2006

Acquisition of quinolone resistance and point mutation of the gyrA gene in Campylobacter jejuni isolated from broilers and in vitro-induced resistant strains.

J Vet Med Sci 2004 Feb;66(2):155-60

Department of Veterinary Public Health, Faculty of Agriculture, Kagoshima University, Japan.

A dramatic rise in the number of resistant Campylobacter to quinolones has been documented in human patients and domestic animals. In this study, the mechanism of acquisition of quinolone resistance was studied by detecting point mutations in the gyrA gene of Campylobacter strains obtained from broilers and strains with in vitro-induced resistance. The minimal inhibitory concentrations (MICs) of norfloxacin (NFLX) and ofloxacin (OFLX) for the strains that had no point mutation were slightly increased from the source strain (Campylobacter jejuni ATCC 33560). The MICs of nalidixic acid (NA), NFLX, and OFLX for the strains that had the point mutation at Thr-86 were 100 or 200 microg/ml, 50 microg/ml, and 25 microg/ml, respectively. The MIC of NA for the strain that had a point mutation at Asp-90 higher than those for the strains that had the point mutation at Thr-86, but the MICs of NFLX and OFLX were relatively lower than those for the strains that had point mutation at Thr-86. These findings suggest that the degree of antimicrobial resistance against NA, NFLX, and OFLX in the in vitro-induced C. jejuni strains was associated with the location of the point mutation in gyrA. On the other hand, a point mutation in all seven resistant strains isolated from broilers was located only at Thr-86, while the MICs of the three quinolones varied in each wild strain. This suggests that another mechanism might also be involved in the acquisition of quinolone resistance in C. jejuni wild strains.
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http://dx.doi.org/10.1292/jvms.66.155DOI Listing
February 2004