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1.
Shokuhin Eiseigaku Zasshi ; 64(4): 117-122, 2023.
Artigo em Japonês | MEDLINE | ID: mdl-37673600

RESUMO

Meat derived from spent hens as well as broilers is destined for human consumption. There are many reports on the prevalence and antimicrobial resistance of Campylobacter and Salmonella in broiler meat, but few in spent hen meat. Therefore, we investigated the prevalence and antimicrobial resistance of these genera in spent hen meat collected at chicken processing plants. Campylobacter and Salmonella were isolated from 47 (92.2%) and 18 (35.5%), respectively, of breast meat derived from 51 spent hen flocks. Campylobacter jejuni accounted for 87.5% of Campylobacter isolates. The highest resistant rate in C. jejuni isolates was found for ampicillin (45.3%), followed by tetracycline (14.3%) and ciprofloxacin (14.3%). There was no Campylobacter isolate resistant to erythromycin, which is recommended as a first-choice antimicrobial for humans when Campylobacter enteritis is strongly suspected. Of Salmonella isolates, the first and second most frequent serovars were Salmonella Corvallis (30.4%) and S. Braenderup (21.7%), respectively. Of Salmonella isolates, 30.4% were resistant to streptomycin. There was no Salmonella isolate resistant to ciprofloxacin, which is one of the recommended antimicrobials for humans against Salmonella enteritis. This study shows that one third of spent hen meat is contaminated with Campylobacter or Salmonella, and administration of erythromycin or cefotaxime is an effective option for patients with Campylobacter- or Salmonella- enteritis, respectively, caused by consumption of spent hen meat.


Assuntos
Campylobacter , Enterite , Animais , Humanos , Feminino , Galinhas , Prevalência , Ciprofloxacina/farmacologia , Eritromicina/farmacologia , Carne , Enterite/epidemiologia
2.
Shokuhin Eiseigaku Zasshi ; 64(6): 214-217, 2023.
Artigo em Japonês | MEDLINE | ID: mdl-38171891

RESUMO

Chicken liver is a potential source of campylobacteriosis in humans. Therefore, we determined the number of Campylobacter in chicken liver. In total, 33 vacuum-packed liver products were obtained from retail stores, and found that 27 of the 33 products (81.8%) were contaminated with Campylobacter. Moreover, Campylobacter was isolated from 138 of 149 livers (92.6%) collected from the 27 Campylobacter-positive products. The mean Campylobacter count was 2.3 log10 CFU/g, while Campylobacter count in 22 of the 138 contaminated livers (15.9%) was >3.0 log10 CFU/g. Furthermore, gastrointestinal tract, liver, and bile samples were collected from 35 broilers at chicken processing plants. We isolated Campylobacter from the gastrointestinal tract of 27 broilers (77.1%). Of these 27 broilers, liver of 24 broilers (88.9%) was Campylobacter-positive, with a mean Campylobacter count of 2.8 log10 CFU/g. Of these 24 broilers, bile of 13 broilers (54.2%) was contaminated with Campylobacter (mean Campylobacter count, 3.5 log10 CFU/mL). Among them, bile of 2 broilers had a Campylobacter count of >8.3 log10 CFU/mL. Collectively, these results indicate that livers derived from broilers colonized with Campylobacter are contaminated with Campylobacter at the time of evisceration. Therefore, to prevent foodborne campylobacteriosis in humans, chicken livers should be thoroughly heated before consumption.


Assuntos
Infecções por Campylobacter , Campylobacter , Animais , Humanos , Infecções por Campylobacter/epidemiologia , Galinhas , Japão , Microbiologia de Alimentos , Carne , Fígado
3.
Bioorg Med Chem Lett ; 43: 128052, 2021 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-33887440

RESUMO

Ciclesonide is an inhaled corticosteroid used to treat asthma and is currently undergoing clinical trials for treatment of coronavirus disease 2019 (COVID-19). An active metabolite of ciclesonide, Cic2, was recently reported to repress severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) genomic RNA replication. Herein, we designed and synthesized a few types of ciclesonide analogues. Cic4 (bearing an azide group) and Cic6 (bearing a chloro group) potently decreased SARS-CoV-2 viral replication and had low cytotoxicity compared with Cic2 (bearing a hydroxy group). These compounds are promising as novel therapeutic agents for COVID-19 that show significant antiviral activity.


Assuntos
Tratamento Farmacológico da COVID-19 , Pregnenodionas/farmacologia , RNA Viral/antagonistas & inibidores , SARS-CoV-2/efeitos dos fármacos , COVID-19/virologia , Glucocorticoides/farmacologia , Humanos , RNA Viral/genética , SARS-CoV-2/genética , Replicação Viral/genética
4.
Foodborne Pathog Dis ; 18(5): 331-336, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33600236

RESUMO

In this study, we aimed to investigate the standard method used for quantification of norovirus in oysters in Japan for the provisional adaptation of the method as an alternative to ISO 15216-1:2017, to conduct a Japan baseline survey of norovirus in oysters. For this purpose, the method provided by the Japan Committee for Standardization of Virus Detection in Food was subjected to an interlaboratory study to determine the performance characteristics of the standard method used in Japan. As a result, the theoretical limit of quantification for norovirus GI and GII in oysters by the standard method used in Japan was expected to be 1.92 and 1.85 log10 copies/g, respectively. The repeatability standard deviations (Sr) were 0.26 and 0.30 log10 copies/g for GI and GII, respectively, and the reproducibility standard deviations (SR) were 0.47 and 0.44 log10 copies/g for GI and GII, respectively. Through the interlaboratory study, we specified several critical points to obtain scientifically reliable results by using the standard method used in Japan. Especially, necessity for application of using process control virus was the most crucial point that needed to be improved. In addition, there are many participating laboratories that could not handle dilution of standard and quantify or detect the viruses in the test samples. To ensure scientifically reliable test result, capacity building of laboratories and implementation of proficiency testing should be considered for future tasks in combination with an application of process control materials in the method. On the assumption that the problems revealed in this study will be solved, the standard method used in Japan would be suitable for use in Japan baseline survey of norovirus in oysters, which will contribute to the international action against norovirus in oysters, led by the EU.


Assuntos
Microbiologia de Alimentos/métodos , Norovirus/genética , Técnicas de Amplificação de Ácido Nucleico/métodos , Ostreidae/virologia , RNA Viral/isolamento & purificação , Animais , Microbiologia de Alimentos/normas , Japão , Técnicas de Amplificação de Ácido Nucleico/normas , Reprodutibilidade dos Testes , Inquéritos e Questionários
5.
Shokuhin Eiseigaku Zasshi ; 61(4): 126-131, 2020.
Artigo em Japonês | MEDLINE | ID: mdl-33012766

RESUMO

Campylobacter is one of the most important causes of food-borne infectious diseases. Antibiotics are rarely needed to treat campylobacteriosis, but occasionally used in severe or prolonged cases. Consumption of contaminated bovine liver is a source of campylobacteriosis. Bovine liver can be contaminated with Campylobacter on the surface and inside by the bile at slaughterhouses. Therefore, we investigated the current prevalence and characteristics of Campylobacter in bovine bile at a slaughterhouse. Campylobacter was isolated from 35.7% (55/154) of bile samples. C. jejuni and C. fetus were the two most frequent species. High antimicrobial resistant rates in C. jejuni were observed against tetracycline (63.0%) and ciprofloxacin (44.4%). Multi-locus sequence typing divided C. jejuni isolates (27 isolates) into 12 sequence types (STs) in which ST806 was the most frequent ST and accounted for 37.0%. All C. fetus were identified as C. fetus subsp. fetus which can cause systemic infections. High antimicrobial resistant rates in C. fetus were observed against ciprofloxacin (66.6%), streptomycin (58.3%) and tetracycline (33.3%). All the C. fetus isolates were divided into two STs, ST3 (16 isolates) and ST6 (8 isolates). Of the 16 ST3 isolates, 15 (93.8%) were resistant to both streptomycin and ciprofloxacin. Our data shows high prevalence of Campylobacter in bovine bile and their high rates of antimicrobial resistance. Preventing bile contamination of bovine liver at slaughterhouses is thus considered to be one of control measures to reduce the risk of Campylobacter infections.


Assuntos
Bile , Campylobacter , Vesícula Biliar , Animais , Anti-Infecciosos/farmacologia , Bile/microbiologia , Campylobacter/classificação , Campylobacter/efeitos dos fármacos , Campylobacter/genética , Campylobacter/isolamento & purificação , Bovinos , Farmacorresistência Bacteriana , Microbiologia de Alimentos , Vesícula Biliar/microbiologia , Tipagem de Sequências Multilocus , Prevalência
6.
J Med Virol ; 91(12): 2101-2107, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31368535

RESUMO

Norovirus (NoV) is a major cause of viral gastroenteritis, and GII.4 has been the predominant genotype worldwide since the mid-1990s. During the 2014 to 2015 winter, a rare genotype, NoV GII.17, emerged and became prevalent mainly in East Asia. Over the past two decades, NoV molecular surveillance in Osaka City, Japan, has revealed that NoV GII.17 was detected for the first time in February 2001 and that NoV GII.17-associated outbreaks remarkably increased during the 2014 to 2015 season, with higher incidence recorded in January to March 2015. Genetic analysis indicated that 28 GII.17 outbreak strains were closely related to the novel GII.P17-GII.17 variants represented by the Kawasaki308/2015/JP strain, similar to that in other regions. Statistical analysis showed that NoV GII.17 infections were more common in adults than GII.3 and GII.4 infections, suggesting that the affected adults most likely did not have antibodies against NoV GII.17 and the novel GII.17 variant had recently appeared. Regarding transmission, food was one of the most important factors involved in the spread of NoV GII.17 among adults; 61% of GII.17 outbreaks were foodborne, with oysters being the most common vehicle. Interplay between pathogens, hosts, and environmental factors was considered to be important in the 2014 to 2015 NoV GII.17 epidemic.


Assuntos
Infecções por Caliciviridae/epidemiologia , Infecções por Caliciviridae/virologia , Surtos de Doenças , Gastroenterite/epidemiologia , Norovirus/genética , Adulto , Animais , Anticorpos Antivirais/sangue , Infecções por Caliciviridae/transmissão , Criança , Cidades/epidemiologia , Doenças Transmitidas por Alimentos/epidemiologia , Doenças Transmitidas por Alimentos/virologia , Gastroenterite/virologia , Genótipo , Humanos , Incidência , Japão/epidemiologia , Ostreidae/virologia , Filogenia , Estações do Ano
7.
Foodborne Pathog Dis ; 15(10): 621-626, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30117743

RESUMO

The contamination of oysters with human norovirus (HuNoV) poses a human health risk, as oysters are often consumed raw. In this study, the effect of high pressure processing (HPP) on a wide variety of HuNoVs naturally present in aqua-cultured Japanese oysters was determined through a polymerase chain reaction-based method with enzymatic pretreatment, to distinguish between infectious HuNoV. Among five batches, genogroup I. genotype 1 (GI.1), GI.2, GI.3, and GI.8 HuNoV were detected from only one oyster not treated with HPP in the fifth batch, while genogroup II. genotype 1 to 4 (GII.1 to 4), GII.6, GII.8., GII.9, GII.13, GII.16, GII.17, and GII.22 HuNoV were detected from oysters not treated with HPP in all tested batches as determined by next-generation sequencing analysis. Neither GI nor GII HuNoV was detected in the oysters of any of the batches after HPP treatment. To our knowledge, this is the first study to investigate the effect of HPP on a wide variety of HuNoVs naturally present in aqua-cultured oysters.


Assuntos
Manipulação de Alimentos , Norovirus/fisiologia , Ostreidae/virologia , Alimentos Marinhos/virologia , Animais , Genótipo , Sequenciamento de Nucleotídeos em Larga Escala , Japão , Norovirus/genética , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real , Frutos do Mar
8.
Foodborne Pathog Dis ; 14(8): 465-471, 2017 08.
Artigo em Inglês | MEDLINE | ID: mdl-28594609

RESUMO

To obtain detailed information on the diversity of infectious norovirus in oysters (Crossostrea gigas), oysters obtained from fish producers at six different sites (sites A, B, C, D, E, and F) in Japan were analyzed once a month during the period spanning October 2015-February 2016. To avoid false-positive polymerase chain reaction (PCR) results derived from noninfectious virus particles, samples were pretreated with RNase before reverse transcription-PCR (RT-PCR). RT-PCR products were subjected to next-generation sequencing to identify norovirus genotypes in oysters. As a result, all GI genotypes were detected in the investigational period. The detection rate and proportion of norovirus GI genotypes differed depending on the sampling site and month. GII.3, GII.4, GII.13, GII.16, and GII.17 were detected in this study. Both the detection rate and proportion of norovirus GII genotypes differed depending on the sampling site and month. In total, the detection rate and proportion of GII.3 were highest from October to December among all detected genotypes. In January, the detection rates of GII.4 and GII.17 reached the same level as that of GII.3. The proportion of GII.17 was relatively lower from October to December, whereas it was the highest in January. To our knowledge, this is the first investigation on noroviruses in oysters in Japan, based on a method that can distinguish their infectivity.


Assuntos
Infecções por Caliciviridae/virologia , Variação Genética , Norovirus/genética , Ostreidae/virologia , Animais , Infecções por Caliciviridae/epidemiologia , Genótipo , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Japão/epidemiologia , Norovirus/isolamento & purificação , Reação em Cadeia da Polimerase , Análise de Sequência de DNA
9.
Foodborne Pathog Dis ; 14(9): 518-523, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-28594572

RESUMO

The contamination of oysters with human noroviruses poses a human health risk, since oysters are often consumed raw. In this study, human norovirus genogroup II was allowed to bio-accumulate in oysters, and then the effect of high-pressure processing (HPP) on human noroviruses in oysters was determined through a polymerase chain reaction (PCR)-based method with enzymatic pretreatment to distinguish infectious noroviruses. As a result, oysters could be artificially contaminated to a detectable level of norovirus genome by the reverse transcription-PCR. Concentrations of norovirus genome in laboratory-contaminated oysters were log normally distributed, as determined by the real-time PCR, suggesting that artificial contamination by bio-accumulation was successful. In two independent HPP trials, a 1.87 log10 and 1.99 log10 reduction of norovirus GII.17 genome concentration was observed after HPP at 400 MPa for 5 min at 25°C. These data suggest that HPP is a promising process of inactivation of infectious human noroviruses in oysters. To our knowledge, this is the first report to investigate the effect of HPP on laboratory-contaminated noroviruses in oysters.


Assuntos
Infecções por Caliciviridae/prevenção & controle , Contaminação de Alimentos/prevenção & controle , Manipulação de Alimentos/métodos , Doenças Transmitidas por Alimentos/prevenção & controle , Norovirus/fisiologia , Ostreidae/virologia , Animais , Infecções por Caliciviridae/virologia , Doenças Transmitidas por Alimentos/virologia , Humanos , Pressão Hidrostática , Reação em Cadeia da Polimerase em Tempo Real
10.
Toxins (Basel) ; 16(6)2024 Jun 04.
Artigo em Inglês | MEDLINE | ID: mdl-38922154

RESUMO

Tetrodotoxin (TTX) is a representative natural toxin causing pufferfish food poisoning, which is especially prominent in East and Southeast Asia, including Japan. TTX has been analyzed through post-column derivatization high-performance liquid chromatography (HPLC), ion-pair LC-MS(/MS), and hydrophilic interaction liquid chromatography (HILIC)-MS(/MS) as alternatives to the mouse bioassay method. However, post-column derivatization requires a system for online derivatization reactions, and with the ion-pair LC-MS approach, it is difficult to remove residual ion-pair reagents remaining in the equipment. Moreover, HILIC-MS provides poor separation compared to reversed-phase (RP) HPLC and requires a long time to reach equilibration. Therefore, we decided to develop a TTX analytical method using pre-column derivatization and RP HPLC for the rapid assessment of outbreak samples, including food remnants. In this study, we focused on the vic-diol moiety of TTX and designed a new derivatization reagent coded as NBD-H-DAB. This NBD-H-DAB was synthesized from 4-hydrazino-7-nitro-2,1,3-benzoxadiazole (NBD-H) and 3-fluoro-2-formylphenylboronic acid (FFPBA) with a simple reaction system and rapidly converted to its boronate form, coded NBD-H-PBA, in an aqueous reaction solution. The NBD-H-PBA demonstrated appropriate hydrophobicity to be retained on the RP analytical column and successfully detected with a UV spectrometer. It was easily reacted with the vic-diol moiety of TTX (C6 and C11) to synthesized a boronic ester. The derivatized TTX could be detected using the RP HPLC-UV, and the limit of detection in the fish flesh samples was 0.06 mg/kg. This novel pre-column derivatization of TTX with NBD-H-PBA proves capable for the analysis of TTX.


Assuntos
Cromatografia de Fase Reversa , Tetrodotoxina , Tetrodotoxina/análise , Tetrodotoxina/química , Animais , Cromatografia Líquida de Alta Pressão , Contaminação de Alimentos/análise , Boro/química , Boro/análise , Espectrometria de Massas em Tandem
11.
Microorganisms ; 11(2)2023 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-36838499

RESUMO

We report for the first time that ephedrine alkaloids-free Ephedra Herb extract (EFE) directly inhibits the replication of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in vitro and that the addition of EFE to the culture medium before viral infection reduces virus titers in the culture supernatant of SARS-CoV-2, including those of variant strains, by more than 99%, 24 h after infection. The addition of Ephedra Herb macromolecule condensed-tannin, which is the main active ingredient responsible for the anticancer, pain suppression, and anti-influenza effects of EFE, similarly suppressed virus production in the culture supernatant by 99% before infection and by more than 90% after infection. Since EFE does not have the side effects caused by ephedrine alkaloids, such as hypertension, palpitations, and insomnia, our results showed the potential of EFE as a safe therapeutic agent against coronavirus disease 2019.

12.
J Vet Med Sci ; 85(4): 463-470, 2023 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-36878553

RESUMO

Understanding the antimicrobial resistance of Campylobacter jejuni and Salmonella spp. isolated from patients with enteritis will aid in therapeutic decision-making. This study aimed to characterize C. jejuni and Salmonella spp. isolates from patients with enteritis. For C. jejuni, the resistance rates against ampicillin, tetracycline, and ciprofloxacin were 17.2%, 23.8%, and 46.4%, respectively. All the C. jejuni isolates were susceptible to erythromycin, which is recommended as a first-choice antimicrobial if Campylobacter enteritis is strongly suspected. C. jejuni was classified into 64 sequence types (STs), and the five major STs were ST22, ST354, ST21, ST918, and ST50. The ciprofloxacin-resistance rate of ST22 was 85.7%. For Salmonella, the resistance rates against ampicillin, cefotaxime, streptomycin, kanamycin, tetracycline, and nalidixic acid were 14.7%, 2.0%, 57.8%, 10.8%, 16.7%, and 11.8%, respectively. All the Salmonella spp. isolates were susceptible to ciprofloxacin. Therefore, fluoroquinolones are the recommended antimicrobials against Salmonella enteritis. S. Thompson, S. Enteritidis, and S. Schwarzengrund were the three most prevalent serotypes. The two cefotaxime-resistant isolates were serotyped as S. Typhimurium and were found to harbor blaCMY-2. The results of this study would help select antimicrobials for treating patients with Campylobacter and Salmonella enteritis.


Assuntos
Anti-Infecciosos , Infecções por Campylobacter , Campylobacter jejuni , Enterite , Animais , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Japão/epidemiologia , Farmacorresistência Bacteriana , Ciprofloxacina/farmacologia , Tetraciclina/uso terapêutico , Infecções por Campylobacter/tratamento farmacológico , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/veterinária , Salmonella , Enterite/epidemiologia , Enterite/veterinária , Anti-Infecciosos/uso terapêutico , Ampicilina/uso terapêutico , Cefotaxima/uso terapêutico , Testes de Sensibilidade Microbiana/veterinária
13.
AIMS Microbiol ; 8(4): 566-574, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36694587

RESUMO

Hepatitis E virus (HEV) is a zoonotic pathogen that circulates mainly between pigs and humans. In Japan, the number of confirmed HEV cases has increased over the past decade, with the majority reported as domestic HEV infections. HEV-infected pork products may be associated with this increase, but there is limited information on HEV in pork in Japanese markets. From February to March 2020, gallbladders were collected from 200 slaughtered pigs shipped from 14 farms and were surveyed to detect HEV RNA in bile using reverse transcription quantitative polymerase chain reaction. The samples were then sequenced and genotyped. Twenty pigs were positive for HEV ribonucleic acid, and seven samples had Ct values of less than 30. Among these 20 pigs, virus strains from 14 pigs were determined as genotype 3. This report indicated that HEV-contaminated pork liver was shipped to consumer markets and demonstrated the importance of detection of HEV in meat ready for shipment.

14.
J Vet Med Sci ; 84(11): 1502-1507, 2022 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-36198611

RESUMO

Campylobacter and non-typhoidal Salmonella are the major causes of bacterial gastrointestinal infections in humans. Although antimicrobial therapy is typically not recommended in many cases of these infections, it may be life-saving in patients with severe symptoms. Since chicken eggs and meat derived from layers are destined for human consumption, we investigated the prevalence and antimicrobial resistance of these two bacterial genera in 82 layer flocks at chicken processing plants in Honshu, Japan. Campylobacter was isolated from 77 flocks (93.9%). Resistance to ampicillin, tetracycline, and ciprofloxacin was documented in 42.3 (30/71), 16.9 (12/71), and 14.1% (10/71) of Campylobacter jejuni, respectively. Multilocus-sequence typing identified ST4389 and ST5262 as the most frequent C. jejuni sequence types. In C. coli, resistance to ampicillin, tetracycline, and ciprofloxacin was found in 20.0 (7/35), 20.0 (7/35), and 25.7% (9/35), respectively. The most frequent sequence type in C. coli was ST8292. Erythromycin resistance was not observed among Campylobacter species. Salmonella was isolated from 14 flocks (17.1%). The two most frequent serovars were Salmonella Corvallis and S. Braenderup. Neither S. Enteritidis nor S. Infantis were isolated. Streptomycin resistance was observed in six isolates (26.1%), and all isolates were susceptible to cefotaxime and ciprofloxacin. Thus, chicken eggs and meat derived from layers are possible sources of these bacterial infections in humans. The antimicrobial susceptibility of these isolates was maintained, reflecting restrictions on the use of antimicrobial agents on layers.


Assuntos
Infecções por Campylobacter , Campylobacter jejuni , Campylobacter , Humanos , Animais , Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Prevalência , Japão/epidemiologia , Testes de Sensibilidade Microbiana/veterinária , Campylobacter jejuni/genética , Salmonella , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/veterinária , Infecções por Campylobacter/microbiologia , Ciprofloxacina/farmacologia , Tetraciclina/farmacologia , Galinhas/microbiologia , Ampicilina
15.
J Vet Med Sci ; 84(6): 743-746, 2022 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-35473799

RESUMO

Chicken is a major source of human campylobacteriosis. Chicken meat originates not only from broilers but also from spent layers; however, few reports have documented the prevalence and antimicrobial resistance of Campylobacter spp. in layers in Japan. Therefore, we investigated the prevalence and antimicrobial susceptibility of Campylobacter spp. in 47 layer farms in Japan. Fecal samples were collected from the youngest and oldest flocks on the farm, and Campylobacter spp. was isolated from 46/47 (97.9%) farms. Among the C. jejuni isolates, the resistance rates to ampicillin, tetracycline, and ciprofloxacin were 29.6%, 22.2%, and 19.8%, respectively. The ciprofloxacin resistance rate (7.3%) in C. jejuni isolated from old flocks was significantly (P<0.01) lower than that in young flocks (32.5%).


Assuntos
Infecções por Campylobacter , Campylobacter coli , Campylobacter jejuni , Animais , Antibacterianos/farmacologia , Infecções por Campylobacter/tratamento farmacológico , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/veterinária , Galinhas , Ciprofloxacina/farmacologia , Farmacorresistência Bacteriana , Fazendas , Fluoroquinolonas/farmacologia , Testes de Sensibilidade Microbiana/veterinária , Prevalência
16.
J Virol ; 84(4): 2110-21, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19955312

RESUMO

Herpesvirus nucleocapsids assemble in the nucleus and must cross the nuclear membrane for final assembly and maturation to form infectious progeny virions in the cytoplasm. It has been proposed that nucleocapsids enter the perinuclear space by budding through the inner nuclear membrane, and these enveloped nucleocapsids then fuse with the outer nuclear membrane to enter the cytoplasm. Little is known about the mechanism(s) for nuclear egress of herpesvirus nucleocapsids and, in particular, which, if any, cellular proteins are involved in the nuclear egress pathway. UL12 is an alkaline nuclease encoded by herpes simplex virus type 1 (HSV-1) and has been suggested to be involved in viral DNA maturation and nuclear egress of nucleocapsids. Using a live-cell imaging system to study cells infected by a recombinant HSV-1 expressing UL12 fused to a fluorescent protein, we observed the previously unreported nucleolar localization of UL12 in live infected cells and, using coimmunoprecipitation analyses, showed that UL12 formed a complex with nucleolin, a nucleolus marker, in infected cells. Knockdown of nucleolin in HSV-1-infected cells reduced capsid accumulation, as well as the amount of viral DNA resistant to staphylococcal nuclease in the cytoplasm, which represented encapsidated viral DNA, but had little effect on these viral components in the nucleus. These results indicated that nucleolin is a cellular factor required for efficient nuclear egress of HSV-1 nucleocapsids in infected cells.


Assuntos
Herpesvirus Humano 1/fisiologia , Nucleocapsídeo/fisiologia , Fosfoproteínas/fisiologia , Proteínas de Ligação a RNA/fisiologia , Transporte Ativo do Núcleo Celular , Animais , Sequência de Bases , Células Cultivadas , Chlorocebus aethiops , Primers do DNA/genética , Desoxirribonucleases/química , Desoxirribonucleases/genética , Desoxirribonucleases/fisiologia , Herpesvirus Humano 1/genética , Herpesvirus Humano 1/ultraestrutura , Microscopia Eletrônica de Transmissão , Complexos Multiproteicos , Fosfoproteínas/antagonistas & inibidores , Fosfoproteínas/química , Fosfoproteínas/genética , RNA Interferente Pequeno/genética , Proteínas de Ligação a RNA/antagonistas & inibidores , Proteínas de Ligação a RNA/química , Proteínas de Ligação a RNA/genética , Coelhos , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Células Vero , Proteínas Virais/química , Proteínas Virais/genética , Proteínas Virais/fisiologia , Montagem de Vírus , Nucleolina
17.
Artigo em Japonês | MEDLINE | ID: mdl-22259842

RESUMO

Human norovirus is the most important foodborne virus in Japan. According to the statistics of food poisoning by the Ministry of Health, Labour, and Welfare (MHLW), the number of patients infected with norovirus has accounted for half of all the patients with food poisoning in recent years. One of the most important measures for the control of infectious diseases is establishing of techniques for inactivating pathogens. For the prevention of food poisoning caused by norovirus, MHLW recommends that foods be subjected to heat treatment at 85 degrees C for 1 min or more; moreover, it recommends the use of sodium hypochlorite to inactivate (disinfect) this virus. However, application of these treatments is not always feasible because heat results in denaturation and sodium hypochlorite can be toxic to the human body and can cause discoloration. Therefore, it is necessary to develop and improve the efficacy of disinfectants and physiochemical treatments against the virus. Human norovirus cannot be propagated in cell culture or in a small animal. This matter is the greatest hindrance for testing the stability of this virus in environments or for evaluating the efficacy of disinfectants, heat treatment, pH treatment, ultraviolet or gamma irradiation, high hydrostatic pressure treatment, and other methods for the inactivation of the virus. Hence, some viruses such as human enterovirus, feline calicivirus, or mouse norovirus have been used as surrogates of human norovirus. The data on inactivation and stability of surrogate viruses are exclusively used as the data of human noroviruses. In recent years, some attempts to distinguish between infectious and noninfectious virus particles by genetic methods such as polymerase chain reaction have been made. These methods include pretreatments by RNase for digesting viral RNAs from non-intact or destroyed virus particles, or addition of a reagent such as ethidium monoazide for inhibiting PCR amplification of viral RNAs from them, before RNA extraction. Non-intact virus particles, which may represent virus particles with some damage (s) in the structural protein(s), are not necessarily synonymous with non-infectious virus particles. However, the results of methods using these treatments, compared to the results of traditional methods without these treatments, seem to be more correlated to the amount of the infectious virus particles. Although many disinfectants or physiochemical treatments have been reported, traditional techniques such as removal of virus particles by washing in running water, heat treatment, or disinfection by sodium hypochlorite are still important control measures. Establishment of control measures for human norovirus and successful propagation of the virus in cell culture are strongly desired.


Assuntos
Microbiologia de Alimentos , Doenças Transmitidas por Alimentos/prevenção & controle , Doenças Transmitidas por Alimentos/virologia , Norovirus/fisiologia , Inativação de Vírus , Animais , Técnicas de Cultura de Células , Desinfetantes , Etanol , Raios gama , Temperatura Alta , Humanos , Concentração de Íons de Hidrogênio , Pressão Hidrostática , Microbolhas , Norovirus/crescimento & desenvolvimento , Norovirus/isolamento & purificação , Norovirus/patogenicidade , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Hipoclorito de Sódio , Ultrassom , Raios Ultravioleta
18.
Biocontrol Sci ; 26(2): 119-125, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34092715

RESUMO

There is a worldwide attempt to develop prevention strategies against SARS-CoV-2 transmission. Here we examined the effectiveness of tungsten trioxide (WO3)-based visible light-responsive photocatalyst on the inactivation of SARS-CoV-2 under different temperatures and exposure durations. The viral titer on the photocatalyst-coated glass slides decreased from 5.93 ± 0.38 logTCID50 /mL to 3.05 ±. 25 logTCID50/mL after exposure to 3,000 lux of the visible light irradiation for 6h at 20℃. On the other hand, lighting without the photocatalyst, or the photocatalyst-coat without lighting retained viral stability. Immunoblotting and electron microscopic analyses showed the reduced amounts of spike protein on the viral surface after the photocatalyst treatment. Our data suggest a possible implication of the photocatalyst on the decontamination of SARS-CoV-2 in indoor environments, thereby preventing indirect viral spread.


Assuntos
COVID-19/transmissão , Catálise/efeitos da radiação , Óxidos/efeitos adversos , SARS-CoV-2/crescimento & desenvolvimento , SARS-CoV-2/patogenicidade , SARS-CoV-2/efeitos da radiação , Tungstênio/efeitos adversos , Inativação de Vírus/efeitos da radiação , Humanos , Luz
19.
J Vet Med Sci ; 83(1): 112-115, 2021 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-33431727

RESUMO

Monitoring the prevalence of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) in pigs could be useful for managing transmission risk to humans. To optimize sampling for LA-MRSA monitoring, we compared the sensitivity of MRSA isolation from skin swabs taken behind the ear and nasal swabs collected from 276 pigs and investigated the prevalence of MRSA in their carcasses. MRSA was isolated from 40 behind the ear skin swabs (14.5%), which was statistically higher than the number isolated from nasal swabs (23 samples, 8.3%). MRSA prevalence in the carcasses was 0.4%. All MRSA isolates were sequence type 398 lineage. Sampling of both the skin behind the ear and nasal mucosa in a pig is recommended to investigate the prevalence of LA-MRSA in pigs.


Assuntos
Staphylococcus aureus Resistente à Meticilina , Infecções Estafilocócicas , Doenças dos Suínos , Matadouros , Animais , Gado , Testes de Sensibilidade Microbiana/veterinária , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/veterinária , Suínos , Doenças dos Suínos/epidemiologia
20.
J Virol ; 83(11): 5773-83, 2009 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-19297494

RESUMO

Us3 is a serine/threonine protein kinase encoded by herpes simplex virus 1 (HSV-1). We recently identified serine at Us3 position 147 (Ser-147) as a physiological phosphorylation site of Us3 (A. Kato, M. Tanaka, M. Yamamoto, R. Asai, T. Sata, Y. Nishiyama, and Y. Kawaguchi, J. Virol. 82:6172-6189, 2008). In the present study, we investigated the effects of phosphorylation of Us3 Ser-147 on regulation of Us3 catalytic activity in infected cells and on HSV-1 pathogenesis. Our results were as follows. (i) Only a small fraction of Us3 purified from infected cells was phosphorylated at Ser-147. (ii) Us3 phosphorylated at Ser-147 purified from infected cells had significantly higher kinase activity than Us3 not phosphorylated at Ser-147. (iii) Phosphorylation of Us3 Ser-147 in infected cells was dependent on Us3 kinase activity. (iv) Replacement of Us3 Ser-147 by alanine significantly reduced viral replication in the mouse cornea and the development of herpes stromal keratitis and periocular skin disease in mice. These results indicated that Us3 catalytic activity is tightly regulated by autophosphorylation of Ser-147 in infected cells and that regulation of Us3 activity by autophosphorylation appeared to play a critical role in viral replication in vivo and in HSV-1 pathogenesis.


Assuntos
Biocatálise , Herpesvirus Humano 1/metabolismo , Herpesvirus Humano 1/patogenicidade , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Virais/metabolismo , Animais , Anticorpos Monoclonais/imunologia , Chlorocebus aethiops , Feminino , Herpesvirus Humano 1/genética , Herpesvirus Humano 1/ultraestrutura , Camundongos , Camundongos Endogâmicos ICR , Microscopia Eletrônica , Mutação/genética , Fosforilação , Proteínas Serina-Treonina Quinases/genética , Serina/genética , Serina/metabolismo , Células Vero , Proteínas Virais/genética , Replicação Viral
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