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1.
Diagn Microbiol Infect Dis ; 95(4): 114883, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31495527

RESUMO

This study reports the first isolation and characterization of a vanD5 genotype vancomycin-resistant Enterococcus faecium strain (E. faecium IPHb306) recovered from a 79-year-old Japanese female inpatient. Species identification was determined by biochemical testing, matrix-assisted laser desorption ionization-time of flight mass spectrometry, and species-specific PCR. Susceptibility tests indicated that E. faecium IPHb306 was resistant to vancomycin but susceptible to teicoplanin. Southern hybridization analyses indicated that E. faecium IPHb306 harbored a vanD5 gene cluster on chromosomal DNA. Growth curve analyses showed that a vancomycin resistance phenotype could be inducible. Sequencing analyses of the vanD5 gene cluster and the ddlE. faecium gene demonstrated several point mutations were present. Because this strain belongs to ST203, a major hospital-adapted lineage, spread of the vanD5 genotype E. faecium ST203 is considered a clinical threat in Japan.

3.
Artigo em Inglês | MEDLINE | ID: mdl-30370825

RESUMO

Background: Efficient, objective measures of mild functional difficulties are lacking. Preliminary data from a novel, non-immersive virtual reality, performance-based task (Virtual Kitchen Challenge; VKC) were obtained to address this gap. Methods: 14 older and 21 younger adults completed cognitive tests and two everyday tasks (breakfast, lunch) in the VKC with virtual objects and a touch-screen and in the Real Kitchen with real objects (order counterbalanced). Automated performance measures were obtained from the VKC program and human coders scored VKC and Real Kitchen videos for errors. Results: Older adults made more errors than younger adults on the VKC and Real Kitchen, with similar error patterns across measures. VKC automated measures were significantly related to measures from human coders, performance on the Real Kitchen, and cognitive test scores. Conclusion: The VKC is a valid and highly efficient performance-based measure of subtle functional difficulties with great potential for future clinical and research applications.

4.
J Food Prot ; 81(9): 1450-1458, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-30080122

RESUMO

The potential human health risk of Japanese ready-to-eat (RTE) foods was investigated by determining the contamination by foodborne bacterial pathogens, the prevalence of opportunistic and nosocomial pathogens, and the antibiotic susceptibility of the isolates recovered from 96 samples of lightly pickled vegetables, 88 samples of Western-style desserts, and 98 samples of RTE fish and seafood products sold at retail in Osaka, Japan. Staphylococcus aureus, including isolates producing staphylococcal enterotoxin (SE), were isolated from six lightly pickled vegetable products, seven Western-style dessert products, and three RTE fish and seafood products. Of these isolates, one SEC-producing isolate from a cake was identified as community-acquired methicillin-resistant S. aureus, which was multilocus sequence type 8 and staphylococcal cassette chromosome mec type IV. Enterobacteriaceae species, including Escherichia coli, Klebsiella oxytoca, Klebsiella pneumoniae, Proteus mirabilis, Serratia marcescens, Citrobacter freundii-Citrobacter braakii, and/or the Enterobacter cloacae complex, were isolated from 92 (95.8%) of the lightly pickled vegetable products, 39 (44.3%) of the Western-style dessert products, and 74 (75.5%) of the RTE fish and seafood products. On the basis of the antimicrobial susceptibility profiles of the opportunistic and nosocomial Enterobacteriaceae pathogens, the third-generation cephalosporin, fosfomycin, and quinolone resistance determinants were investigated. We detected AmpC products of the CIT group and a qnrB9 product in 5 and 1 C. freundii-C. braakii isolates, respectively, and fosA gene products in 15 E. cloacae complex isolates. Because RTE foods are consumed without a heating process, the spread of bacterial pathogens from contaminated food to human consumers is possible. RTE foods must be handled using hygienic procedures from the processing steps to the table to reduce the prevalence of potentially pathogenic or pathogenic bacteria and to prevent bacterial growth.


Assuntos
Anti-Infecciosos , Fast Foods/microbiologia , Contaminação de Alimentos/análise , Staphylococcus aureus Resistente à Meticilina , Animais , Antibacterianos , Microbiologia de Alimentos , Humanos , Japão , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Prevalência
5.
J Food Prot ; 81(8): 1346-1350, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-30019957

RESUMO

Kudoa iwatai, a myxosporean parasite, has low host fish specificity, and consumers encounter commercial marine fish or marketed marine fish infected with this parasite in Japan. Although the presence of this parasite infection in fish samples is traditionally determined by the microscopic morphological examination of extracted spores, this method lacks sensitivity and specificity. In this study, we developed a real-time PCR assay for the detection of K. iwatai 18S rDNA to achieve the rapid and specific identification of K. iwatai in foreign substance inspection. We also evaluated the usefulness of real-time PCR for Japanese seabass ( Lateolabrax japonicus) with or without K. iwatai cysts. Our real-time PCR assay was able to reliably detect the target plasmid DNA over a 7-log range (from 4.0 × 101 to 4.0 × 107 copies per reaction) and displayed a linear relationship, with a correlation of determination value of 0.9993 and slope of -3.3651. Moreover, the mean value of the intra-assay coefficient of variation was 0.89% in triplicate assays, and the detection limit of this method was 2.5 copies of K. iwatai 18S rDNA per reaction. The sensitivity of the real-time PCR was the same or higher than that of an established conventional PCR when DNA extracts from eight Japanese seabass with or without K. iwatai were used as templates. The specificity of the real-time PCR was comparable with that of conventional PCR by using DNA extracts from fish samples infected with nine Kudoa species. Together, these results indicate that our real-time PCR assay is highly sensitive, reproducible, and specific for detecting K. iwatai 18S rDNA in foreign substance inspection. We believe that this highly sensitive real-time PCR may also be useful for understanding the gastrointestinal diseases associated with K. iwatai and for studying the yet unknown life cycle of K. iwatai.


Assuntos
Bass/parasitologia , Parasitologia de Alimentos , Myxozoa , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , Aquicultura , Japão , Myxozoa/genética , Myxozoa/isolamento & purificação , Doenças Parasitárias em Animais/parasitologia , Filogenia , RNA Ribossômico 18S , Análise de Sequência de DNA
6.
Support Care Cancer ; 26(9): 3217-3223, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29626261

RESUMO

PURPOSE: We aimed to investigate the relationship between Borg scale and intensity of resistance training in patients who had undergone allogeneic hematopoietic stem cell transplantation (allo-HSCT). Furthermore, the relationship between Borg scale, heart rate (HR), and intensity of exercise tolerance test was also studied. METHODS: The study included 28 patients (19 men and 9 women) who had undergone allo-HSCT between June 2015 and February 2017. Their knee extension strengths and exercise tolerances were evaluated. Patients were asked to grade between 0 and 10 on Borg scale based on the level of difficulty experienced during exercising, after 10 repetitions in randomized 20, 40, and 60% resistance training for knee extension. Additionally, we evaluated Borg scale, HR, and load intensity during exercise tolerance test, every minute of the exercise for 2 weeks before and 3 weeks after HSCT. RESULTS: Knee extension strength and exercise tolerance were significantly decreased 3 weeks after HSCT from those before HSCT (p < 0.01). Additionally, rise in Borg scale with increase in load intensity during knee extension resistance training, both before and after HSCT (p < 0.01), was noted. Furthermore, Borg scale was found to be associated with HR and load intensity during exercise tolerance test in patients both before and after HSCT (p < 0.01). CONCLUSIONS: A correlation was found between Borg scale with intensity of resistance training and exercise tolerance in patients who had undergone allo-HSCT. Therefore, Borg scale could be useful to determine the intensity of physical exercise in patients who have undergone allo-HSCT.


Assuntos
Terapia por Exercício/métodos , Tolerância ao Exercício/fisiologia , Transplante de Células-Tronco Hematopoéticas/métodos , Treinamento de Resistência/métodos , Condicionamento Pré-Transplante/métodos , Transplante Homólogo/métodos , Adolescente , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Adulto Jovem
7.
J Clin Microbiol ; 56(6)2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29593058

RESUMO

In Escherichia coli, more than 180 O groups and 53 H types have been recognized. The O:H serotyping of E. coli strains is an effective method for identifying strains with pathogenic potential and classifying them into clonal groups. In particular, the serotyping of Shiga toxin-producing E. coli (STEC) strains provides valuable information to evaluate the routes, sources, and prevalence of agents in outbreak investigations and surveillance. Here, we present a complete and practical PCR-based H-typing system, E. coli H-genotyping PCR, consisting of 10 multiplex PCR kits with 51 single PCR primer pairs. Primers were designed based on a detailed comparative analysis of sequences from all H-antigen (flagellin)-encoding genes, fliC and its homologs. The specificity of this system was confirmed by using all H type reference strains. Additionally, 362 serotyped wild strains were also used to evaluate its practicality. All 277 H-type-identified isolates gave PCR products that corresponded to the results of serological H typing. Moreover, 76 nonmotile and nine untypeable strains could be successfully subtyped into any H type by the PCR system. The E. coli H-genotyping PCR developed here allows broader, rapid, and low-cost subtyping of H types and will assist epidemiological studies as well as surveillance of pathogenic E. coli.


Assuntos
Antígenos de Bactérias/genética , Escherichia coli/classificação , Técnicas de Genotipagem , Tipagem Molecular/métodos , Reação em Cadeia da Polimerase Multiplex/métodos , Primers do DNA , DNA Bacteriano/genética , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/diagnóstico , Proteínas de Escherichia coli/genética , Flagelina/genética , Genótipo , Humanos , Tipagem Molecular/economia , Sorogrupo
8.
J Clin Microbiol ; 56(5)2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29491014

RESUMO

In September 2016, 140 patients with primary symptoms of sore throat and fever were identified in a school dormitory in Osaka, Japan. Epidemiological and laboratory investigations determined that these symptomatic conditions were from a foodborne outbreak of group G streptococcus (GGS), with GGS being isolated from samples from patients, cooks, and foods. The strain of GGS was identified as Streptococcus dysgalactiae subsp. equisimilis of two emm types (stG652.0 and stC36.0). The causative food, a broccoli salad, was contaminated with the two types of S. dysgalactiae subsp. equisimilis, totaling 1.3 × 104 CFU/g. Pulsed-field gel electrophoresis (PFGE) of samples from patients, cooks, and foods produced similar band patterns among samples with the same emm type. This result suggested the possibility of exposure from the contaminated food. The average onset time was 44.9 h and the prevalence rate was 62%. This is the first report to identify the causative food of a foodborne outbreak by Streptococcus dysgalactiae subsp. equisimilis.


Assuntos
Surtos de Doenças , Microbiologia de Alimentos , Faringite/epidemiologia , Instituições Acadêmicas , Infecções Estreptocócicas/epidemiologia , Streptococcus/isolamento & purificação , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/genética , Proteínas da Membrana Bacteriana Externa/genética , Brassica/microbiologia , Proteínas de Transporte/genética , DNA Bacteriano/genética , Eletroforese em Gel de Campo Pulsado , Humanos , Japão/epidemiologia , Faringite/diagnóstico , Faringite/patologia , Instituições Residenciais , Infecções Estreptocócicas/diagnóstico , Infecções Estreptocócicas/patologia , Streptococcus/genética , Streptococcus/crescimento & desenvolvimento , Streptococcus/imunologia
9.
Int J Food Microbiol ; 259: 59-67, 2017 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-28822890

RESUMO

Kudoa septempunctata, a myxosporean parasite of the olive flounder (Paralichthys olivaceus), causes foodborne gastroenteritis after ingestion of contaminated raw flounder. Available methods to detect K. septempunctata require expensive equipment, well-trained personnel, and lengthy procedures. Here we generated a novel monoclonal antibody (MAb 15G11) against K. septempunctata and used it to produce a prototype immunochromatographic assay (prototype Kudoa-ICA). Within 15min, the prototype Kudoa-ICA detected ≥1.0×105spores/mL in a spore suspension and ≥2.0×104spores/g of P. olivaceus muscle. The prototype Kudoa-ICA weakly cross-reacted with spores of K. lateolabracis and K. iwatai. cDNA sequence, expression, and western blot analyses revealed that MAb 15G11 detected an approximately 24-kDa protein encoded by a 573bp mRNA. The cDNA nucleotide and predicted amino acid sequences were not significantly similar to any sequence in the GeneBank database. Immunoelectron microscopy revealed that MAb 15G11 reacted with the sporoplasmic cells and mainly with the capsulogenic cells of the K. septempunctata spore. Although the Kudoa-ICA was weakly cross-reactive with two other Kudoa species, it detected >1.0×106spores/g of K. septempunctata in P. olivaceus muscle, which is the criterion used to indicate a violation of the Food Hygiene Law of Japan. We conclude that MAb 15G11 may be suitable for use in an immunochromatographic assay for screening P. olivaceus muscle contaminated with K. septempunctata at food distribution sites such as food wholesalers, grocery stores, and restaurants.


Assuntos
Anticorpos Monoclonais/imunologia , Cromatografia de Afinidade/métodos , Linguado/parasitologia , Doenças Transmitidas por Alimentos/prevenção & controle , Gastroenterite/prevenção & controle , Myxozoa/imunologia , Esporos de Protozoários/imunologia , Sequência de Aminoácidos/genética , Animais , Anticorpos Monoclonais/genética , Sequência de Bases , Doenças dos Peixes/parasitologia , Doenças Transmitidas por Alimentos/parasitologia , Gastroenterite/parasitologia , Japão , Músculos/parasitologia , Myxozoa/genética , Myxozoa/isolamento & purificação , Esporos de Protozoários/isolamento & purificação
10.
J Food Prot ; : 716-724, 2017 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-28350183

RESUMO

To investigate the microbial quality of retail pepper in Vietnam, the enumeration and detection of Enterobacteriaceae and the screening of cefotaxime (CTX)-resistant coliforms were performed by using 84 commercial samples. Although Enterobacteriaceae were isolated from 78 samples, the number of Enterobacteriaceae was lower than 1.0 log CFU/g in 46 samples. For the detection of Enterobacteriaceae with the International Organization for Standardization methods, Salmonella spp., Escherichia coli , Klebsiella pneumoniae , Cronobacter sakazakii , and Enterobacter cloacae complex were isolated from 5, 12, 36, 19, and 30 samples, respectively. During screening of CTX-resistant coliforms, K. pneumoniae , C. sakazakii , and E. cloacae complex were isolated from 8, 1, and 21 samples, respectively. Seven K. pneumoniae and seven E. cloacae complex isolates obtained in the screening of CTX-resistant coliforms were resistant to at least one of the three third-generation cephalosporins (CTX, ceftazidime, and cefpodoxime). Moreover, one E. cloacae complex cluster IV and all K. pneumoniae isolates were positive for extended-spectrum ß-lactamase genes or plasmid-mediated AmpC ß-lactamase genes or both. Additionally, two extended-spectrum ß-lactamase-producing K. pneumoniae isolates and one AmpC ß-lactamase-producing E. cloacae complex cluster IV isolate were positive for the plasmid-mediated quinolone resistance determinants and also had amino acid alterations in the quinolone resistance-determining regions of GyrA and ParC. Furthermore, 10 E. cloacae complex isolates were positive for the plasmid-mediated fosfomycin resistance gene fosA. As pepper is often consumed without a heating process, the possible spread to humans of foodborne, opportunistic, and nosocomial infection pathogens or resistance genes from foods prepared or seasoned with pepper cannot be excluded. Therefore, it is necessary to handle pepper by using hygienic conditions during the cultivation, harvesting and processing steps.

11.
Int J Syst Evol Microbiol ; 66(10): 3779-3785, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27374383

RESUMO

Two Gram-stain-positive strains, VE80T and VE116, which were resistant to vancomycin, were isolated from retail chicken meat and liver in Ho Chi Minh, Vietnam, respectively. These strains were characterized by sequence analyses of 16S rRNA, RNA polymerase α-subunit (rpoA), ATP synthase α-subunit (atpA), and phenylalanyl-tRNA synthase α-subunit (pheS) genes, determination of DNA G+C content, cellular fatty acid methyl ester analysis, DNA-DNA hybridization, and conventional morphological and biochemical tests. Strains VE80T and VE116 had 99.6 % 16S rRNA gene sequence similarity with Enterococcus canintestini LMG 13590T, and 99.1 % 16S rRNA gene sequence similarity with Enterococcus dispar ATCC 51266T. However, the two isolates could be clearly differentiated from these reference strains by the low sequence similarities (86.1-86.8 %) of the atpA gene, low DNA-DNA relatedness (<22.8 %), and differences in the production of acid from melezitose and methyl α-d-glucoside. Based on the results obtained in the present study, these two isolates are considered to represent a novel species of the genus Enterococcus, for which the name Enterococcus saigonensis sp. nov., is proposed. The type strain is VE80T (=JCM 31193T=CCUG 68827T).


Assuntos
Galinhas/microbiologia , Enterococcus/classificação , Fígado/microbiologia , Carne/microbiologia , Filogenia , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , RNA Polimerases Dirigidas por DNA/genética , Enterococcus/genética , Enterococcus/isolamento & purificação , Ácidos Graxos/química , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vietnã
12.
J Food Prot ; 78(10): 1800-11, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26408128

RESUMO

Shiga toxin family members have recently been classified using a new nomenclature into three Stx1 subtypes (Stx1a, Stx1c, and Stx1d) and seven Stx2 subtypes (Stx2a, Stx2b, Stx2c, Stx2d, Stx2e, Stx2f, and Stx2g). To develop screening methods for Stx genes, including all of these subtype genes, and Escherichia coli O26-, O111-, and O157-specific genes in laboratory investigations of Shiga toxin-producing E. coli (STEC) foodborne cases, we developed multiplex real-time PCR assays and evaluated their specificity and quantitative accuracy using STEC and non-STEC isolates, recombinant plasmids, and food enrichment cultures and by performing STEC spiking experiments with beef and sprout enrichment cultures. In addition, we evaluated the relationship between the recovery rates of the target strains by direct plating and immunomagnetic separation and the cycle threshold (CT) values of the real-time PCR assays for the Stx subtypes and STEC O26, O111, and O157 serogroups. All three stx1- and seven stx2-subtype genes were detected by real-time PCR with high sensitivity and specificity, and the quantitative accuracy of this assay was confirmed using control plasmids and STEC spiking experiments. The results of the STEC spiking experiments suggest that it is not routinely possible to isolate STEC from enrichment cultures with real-time PCR CT values greater than 30 by direct plating on MacConkey agar, although highly selective media and immunomagnetic beads were able to isolate the inoculated strains from the enrichment cultures. These data suggest that CT values obtained from the highly quantitative real-time PCR assays developed in this study provide useful information to develop effective isolation strategies for STEC from food samples. The real-time PCR assays developed here are expected to aid in investigations of infections or outbreaks caused by STEC harboring any of the stx-subtype genes in the new Stx nomenclature, as well as STEC O26, O111, and O157.


Assuntos
Carne Vermelha/microbiologia , Plântula/microbiologia , Toxina Shiga I/isolamento & purificação , Toxina Shiga II/isolamento & purificação , Escherichia coli Shiga Toxigênica/genética , Animais , Sequência de Bases , Bovinos , Contaminação de Alimentos/análise , Microbiologia de Alimentos , Humanos , Separação Imunomagnética/métodos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase Multiplex/métodos , Toxina Shiga I/genética , Toxina Shiga II/genética , Escherichia coli Shiga Toxigênica/classificação , Escherichia coli Shiga Toxigênica/isolamento & purificação
13.
J Phys Ther Sci ; 26(8): 1283-6, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25202198

RESUMO

[Purpose] VO2 is expressed as the product of cardiac output and O2 extraction by the Fick equation. During the incremental exercise test and constant high-intensity exercise test, VO2 results in the attainment of maximal O2 uptake at exhaustion. However, the differences in the physiological components, cardiac output and muscle O2 extraction, have not been fully elucidated. We tested the hypothesis that constant exercise would result in higher O2 extraction than incremental exercise at exhaustion. [Subjects] Twenty-five subjects performed incremental exercise and constant exercise at 80% of their peak work rate. [Methods] Ventilatory, cardiovascular, and muscle oxygenation responses were measured using a gas analyzer, Finapres, and near-infrared spectroscopy, respectively. [Results] VO2 was not significantly different between the incremental exercise and constant exercise. However, cardiac output and muscle O2 saturation were significantly lower for the constant exercise than the incremental exercise at the end of exercise. [Conclusion] These findings indicate that if both tests produce a similar VO2 value, the VO2 in incremental exercise would have a higher ratio of cardiac output than constant exercise, and VO2 in constant exercise would have a higher ratio of O2 extraction than incremental exercise at the end of exercise.

14.
Food Chem ; 162: 94-8, 2014 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-24874362

RESUMO

Using a streptavidin-coated well plate, a biotin-labelled anti-gonyautoxin 2/3 monoclonal antibody GT-13A, and a decarbamoyl saxitoxin-peroxidase conjugate, a direct competitive enzyme-linked immunosorbent assay (PSP-ELISA) was developed for monitoring paralytic shellfish poisoning (PSP) toxins in shellfish. This assay is simple to perform and can be completed in approximately 20 min. The PSP-ELISA was compared to the mouse bioassay (MBA) for the detection of PSP toxins in shellfish samples (n=83) collected from the coast of Osaka Prefecture, Japan. When positive and negative results were indicated based on the regulatory limit for PSP toxins (4 mouse unit(MU)/g of shellfish meat), the PSP-ELISA results showed a sensitivity of 100% (25 of 25) and a specificity of 89.7% (52 of 58 samples) compared to the MBA results. These results suggest that the PSP-ELISA could be used as a rapid and simple screening method prior to the MBA.


Assuntos
Bioensaio/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Intoxicação por Frutos do Mar/diagnóstico , Frutos do Mar/efeitos adversos , Animais , Camundongos , Frutos do Mar/análise
15.
Infect Immun ; 82(6): 2390-9, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24664508

RESUMO

Clostridium perfringens is a causative agent of food-borne gastroenteritis for which C. perfringens enterotoxin (CPE) has been considered an essential factor. Recently, we experienced two outbreaks of food-borne gastroenteritis in which non-CPE producers of C. perfringens were strongly suspected to be the cause. Here, we report a novel enterotoxin produced by C. perfringens isolates, BEC (binary enterotoxin of C. perfringens). Culture supernatants of the C. perfringens strains showed fluid-accumulating activity in rabbit ileal loop and suckling mouse assays. Purification of the enterotoxic substance in the supernatants and high-throughput sequencing of genomic DNA of the strains revealed BEC, composed of BECa and BECb. BECa and BECb displayed limited amino acid sequence similarity to other binary toxin family members, such as the C. perfringens iota toxin. The becAB genes were located on 54.5-kb pCP13-like plasmids. Recombinant BECb (rBECb) alone had fluid-accumulating activity in the suckling mouse assay. Although rBECa alone did not show enterotoxic activity, rBECa enhanced the enterotoxicity of rBECb when simultaneously administered in suckling mice. The entertoxicity of the mutant in which the becB gene was disrupted was dramatically decreased compared to that of the parental strain. rBECa showed an ADP-ribosylating activity on purified actin. Although we have not directly evaluated whether BECb delivers BECa into cells, rounding of Vero cells occurred only when cells were treated with both rBECa and rBECb. These results suggest that BEC is a novel enterotoxin of C. perfringens distinct from CPE, and that BEC-producing C. perfringens strains can be causative agents of acute gastroenteritis in humans. Additionally, the presence of becAB on nearly identical plasmids in distinct lineages of C. perfringens isolates suggests the involvement of horizontal gene transfer in the acquisition of the toxin genes.


Assuntos
Clostridium perfringens/metabolismo , Enterotoxinas/metabolismo , Gastroenterite/microbiologia , ADP Ribose Transferases/genética , Doença Aguda , Análise de Variância , Animais , Modelos Animais de Doenças , Surtos de Doenças , Enterotoxinas/genética , Humanos , Camundongos , Peso Molecular , Coelhos , Proteínas Recombinantes/metabolismo , Análise de Sequência de DNA
16.
Jpn J Infect Dis ; 66(6): 530-3, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24270144

RESUMO

We describe our laboratory investigation of a massive foodborne outbreak of gastrointestinal illness caused by enterotoxigenic Escherichia coli (ETEC) serotype O169:H41 that occurred during a 2-day traditional festival held in September 2012 in Osaka Prefecture, Japan. Of 126 customers who patronized a particular Japanese restaurant during the event, 102 developed symptoms of gastrointestinal disease. We isolated strains of ETEC serotype O169:H41 from 1 food sample and from fecal samples collected from 19 of 34 patients and 2 of 4 food handlers. Pulsed-field gel electrophoresis analysis of these isolates suggested that the foodborne pathogen that caused the diarrheal outbreak was a specific clone of ETEC serotype O169:H41. Based on these findings and our interviews with the restaurant owner and employees, we concluded that a likely cause of the outbreak was an overwhelmed capacity of the restaurant kitchen in terms of preservation of sanitary procedures during the festival and the inability of the restaurant staff to handle the relatively large quantity of food to ensure a lack of contamination with ETEC. Thus, we reconfirm that ETEC strains of serotype O169:H41 remain important causes of domestic foodborne outbreaks in developed countries, including Japan.


Assuntos
Surtos de Doenças , Escherichia coli Enterotoxigênica/isolamento & purificação , Infecções por Escherichia coli/microbiologia , Doenças Transmitidas por Alimentos/microbiologia , Gastroenteropatias/microbiologia , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Escherichia coli Enterotoxigênica/classificação , Escherichia coli Enterotoxigênica/genética , Infecções por Escherichia coli/epidemiologia , Feminino , Doenças Transmitidas por Alimentos/epidemiologia , Gastroenteropatias/epidemiologia , Humanos , Lactente , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Filogenia , Adulto Jovem
17.
Diagn Microbiol Infect Dis ; 77(2): 176-8, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23891550

RESUMO

We isolated an Escherichia coli O157:H7 strain that was negative for verocytotoxin production, but positive for the vtx2 gene using commercial kits, from an asymptomatic food handler. The laboratory investigations revealed that a 1310-bp insertion sequence, IS1203 variant, was present in the B subunit-coding region of the vtx2c gene.


Assuntos
Portador Sadio/microbiologia , Infecções por Escherichia coli/microbiologia , Escherichia coli O157/genética , Genes Bacterianos , Mutagênese Insercional , Toxinas Shiga/genética , Adulto , DNA Bacteriano/análise , DNA Bacteriano/genética , Escherichia coli O157/patogenicidade , Feminino , Manipulação de Alimentos , Humanos , Adulto Jovem
19.
J Food Prot ; 75(10): 1774-82, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23043825

RESUMO

To determine the prevalence and antimicrobial susceptibility profiles of Campylobacter, Salmonella, Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), and vancomycin-resistant enterococci (VRE) in food-producing animals and retail raw meats in Japan, raw meat samples as well as food-producing animal feces, cutaneous swabs, and nasal swabs collected from 2004 to 2006 were analyzed. Isolation rates of Campylobacter jejuni and Campylobacter coli, Salmonella, and S. aureus were 34.6% (363 of 1,050), 2.7% (28 of 1,050), and 32.8% (238 of 725), respectively. MRSA was isolated from 3% (9 of 300) of meat samples. No VRE were isolated in this study. Antibiotic resistance in C. coli was higher than that in C. jejuni. Three C. jejuni isolates from a patient with diarrhea in a hospital of Shizuoka Prefecture and two chicken samples that exhibited resistance to ciprofloxacin had identical pulsed-field gel electrophoresis patterns, suggesting that ciprofloxacin-resistant C. jejuni could have been distributed in meat. S. aureus isolates showed the highest level of resistance to ampicillin and tetracycline. Resistance to tetracycline in S. aureus isolates from beef was lower than that seen in isolates from chicken and pork (P < 0.01). This study revealed that the prevalence of MRSA and VRE were low in food-producing animals and retail domestic meats in Japan, although Campylobacter isolates resistant to fluoroquinolone and erythromycin were detected. The occurrence of antimicrobial-resistant pathogens should be monitored continuously to improve the management of the risks associated with antimicrobial drug resistance transferred from food-producing animals to humans.


Assuntos
Animais Domésticos/microbiologia , Antibacterianos/farmacologia , Contagem de Colônia Microbiana/métodos , Farmacorresistência Bacteriana , Carne/microbiologia , Animais , Campylobacter/isolamento & purificação , Bovinos/microbiologia , Galinhas/microbiologia , Qualidade de Produtos para o Consumidor , Enterococcus/isolamento & purificação , Humanos , Japão , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Testes de Sensibilidade Microbiana , Salmonella/isolamento & purificação , Staphylococcus aureus/isolamento & purificação , Suínos/microbiologia , Resistência a Vancomicina
20.
J Clin Microbiol ; 50(9): 2964-8, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22760033

RESUMO

Kudoa septempunctata is a newly identified myxosporean parasite of olive flounder (Paralichthys olivaceus) and a suspected causative agent of several food-borne gastroenteritis outbreaks in Japan. Here, we report the detection of K. septempunctata 18S ribosomal DNA in fecal samples of outbreak patients using an efficient method based on real-time PCR. We first performed a spiking experiment to assess whether our previously developed real-time PCR assay was applicable to detect K. septempunctata in feces. Simultaneously, we compared the relative extraction efficacy of K. septempunctata DNA using three commercial kits. Finally, our detection method was validated by testing 45 clinical samples obtained from 13 food-borne outbreaks associated with the consumption of raw flounder and 41 fecal samples from diarrhea patients epidemiologically unrelated to the ingestion of raw fish. We found that the FastDNA Spin Kit for Soil (MP Biomedicals) was the most efficient method for extracting K. septempunctata DNA from fecal samples. Using this kit, the detection limit of our real-time PCR assay was 1.6 × 10(1) spores per g of feces, and positive results were obtained for 21 fecal and 2 vomitus samples obtained from the food-borne outbreaks. To our knowledge, this is the first report to describe the detection of K. septempunctata DNA in patient fecal samples. We anticipate that our detection method will be useful for confirming food-borne diseases caused by K. septempunctata in laboratory investigations.


Assuntos
Surtos de Doenças , Fezes/parasitologia , Doenças Transmitidas por Alimentos/epidemiologia , Myxozoa/isolamento & purificação , Doenças Parasitárias/epidemiologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , DNA Ribossômico/genética , DNA Ribossômico/isolamento & purificação , Linguado/parasitologia , Microbiologia de Alimentos , Doenças Transmitidas por Alimentos/parasitologia , Humanos , Japão , Myxozoa/genética , Doenças Parasitárias/parasitologia , RNA Ribossômico 18S/genética , Manejo de Espécimes/métodos
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