Publications by authors named "Hiroshi Asakura"

94 Publications

Bacterial Distribution and Community Structure in Beef Cattle Liver and Bile at Slaughter.

J Food Prot 2021 Nov 24. Epub 2021 Nov 24.

National Institute of Health Sciences 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki, JAPAN Kanagawa 210-9501 +81442706563.

In this study, the distribution of hygienic indicator bacteria in cattle livers and bile was examined at slaughterhouses. First, 127 cattle livers with gallbladders were carefully eviscerated from the carcasses at 10 slaughterhouses. Microbiological examination showed that 9 bile (7.1%) and 19 liver parenchyma (15.0%) samples were positive for the family Enterobacteriaceae (EB) with means ± SD of 3.68 ± 4.63 log CFU/mL and 1.59 ± 2.47 log CFU/g, respectively; thus, bacterial contamination was apparent even at the postevisceration stage. Subsequently, 70 cattle livers were obtained at the postprocessing/storage stage from 7 of the ten slaughterhouses; microbiological analysis revealed greater means of EB in the liver parenchyma (means ± SD of 3.00 ± 3.89 log CFU/g, P =0.011) than those at postevisceration stage, suggesting that bacterial dissemination and/or replication occurred in the liver parenchyma during processing and storage. According to 16S rRNA ion semiconductor sequencing analysis of representative samples from 12 cattle, Proteobacteria , Firmicutes , and Actinobacteria were dominant in both the parenchyma and bile, in which EB/ Escherichia coli were predominate among EB-rich livers. These results suggest that bile plays a role as a vehicle for bacterial transmission to the liver parenchyma. This is the first study to demonstrate bacterial distribution and community structure in the liver and biliary microecosystem of cattle at slaughter. Our data provide possible implication of EB testing in bile to screen cattle livers contaminated with high levels of fecal indicator bacteria.
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http://dx.doi.org/10.4315/JFP-21-288DOI Listing
November 2021

[Analysis of Ciguatoxins in Variola louti Captured off the Ogasawara (Bonin) Islands].

Shokuhin Eiseigaku Zasshi 2021 ;62(5):157-161

National Institute of Health Sciences.

Ciguatera poisoning (CP) is one of the most abundant seafood poisonings in the world. CP frequently occurred in the tropical and subtropical Indo-Pacific Ocean and the Caribbean Sea. In Japan, CP cases have been reported annually, from the subtropical regions, including Okinawa Prefecture and Amami Islands, Kagoshima Prefecture. The principal toxins, named ciguatoxins (CTXs), are bio-synthesized by benthic dinoflagellate of genera Gambierdiscus and Fukuyoa. They are bio-transferred herbivorous animals to carnivorous fishes via the food chain.The Ogasawara Islands comprise more than 30 islands, Mukojima Islands, Chichijima (Bonin) Islands, Hahajima Islands, Iwo Islands, Nishinoshima, Minamitorishima, and Okinotorishima, which locate in the tropical to subtropical regions. The Mukojima Islands, Chichijima Islands, and Hahajima Islands locate approximately the same latitude as Okinawa. The distance from Tokyo is approximately 1,000 km for Chichijima, 1,700 km for Okinotorishima (the southernmost tip of Japan), and 1,900 km for Minamitorishima (the easternmost tip of Japan). These islands exist in a wide range of waters, latitudes from 20°25' to 27°44' North and longitudes from 136°04' to 153° 59' East. We collected 65 specimens of a grouper, Variola louti, the most frequent species implicated in CP in Japan, from the waters around the Chichijima, Mukojima, and Hahajima islands. The fish flesh specimens were analyzed CTXs using the liquid chromatography-tandem mass spectrometer (LC-MS/MS). While the peak whose retention time is almost identical to that of CTX1B was detected in all specimens on our routine protocol, no 52-epi-54-deoxyCTX1B nor 54-deoxyCTX1B was detected. The peak retention time was quite different from that of CTX1B when re-analyzing by changing the analytical column. Thus, the CTXs in the specimens in the waters of these islands seemed to be undetectable levels.
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http://dx.doi.org/10.3358/shokueishi.62.157DOI Listing
November 2021

Development and Evaluation of Fluorescence Immunochromatography for Rapid and Sensitive Detection of Thermophilic .

Food Saf (Tokyo) 2021 Sep 24;9(3):81-87. Epub 2021 Sep 24.

Bacteriology Section, Osaka Institute of Public Health, 1-3-69 Nakamichi, Higashinari-ku, Osaka 537-0025, Japan.

() and () are leading causes of foodborne gastroenteritis in Japan. Epidemiological surveillance has provided evidence that poultry meat is one of the main reservoirs for human campylobacteriosis, and therefore, improvement in process hygiene at slaughter is required to reduce the number of human infections. This study thus aimed to develop fluorescent immunochromatography strips for rapid and sensitive detection of thermophilic on poultry carcasses at slaughter. To establish the required detection levels, we first determined the numbers of and on poultry carcasses at one large-scale poultry slaughterhouse in Japan, resulting in the detection of at 1.97 ± 0.24 log CFU/25 g of neck skin during the post-chilling process by using ISO 10272-2:2017. Our developed fluorescence immunochromatography (FIC) assay exhibited a 50% limit of detection of 3.51 log CFU or 4.34 log CFU for NCTC 11168 or JCM 2529, respectively. Inclusive and exclusive tests resulted in good agreement. The practical usefulness of this test toward poultry carcasses should be evaluated in future studies, perhaps concentration of the target microorganisms prior to the testing might be helpful to further enhance sensitivity. Nevertheless, our data suggest the potential of FIC for rapid and sensitive detection of thermophilic for monitoring the process hygiene of poultry carcasses at slaughter.
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http://dx.doi.org/10.14252/foodsafetyfscj.D-21-00006DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8472094PMC
September 2021

Whole-genome sequence analysis of Shiga toxin-producing Escherichia coli O157 strains isolated from wild deer and boar in Japan.

J Vet Med Sci 2021 Oct 8. Epub 2021 Oct 8.

Laboratory of Veterinary Food Hygiene, Department of Veterinary Medicine, College of Bioresource Sciences, Nihon University.

The prevalence of Shiga toxin-producing Escherichia coli O157 (STEC O157) strains in wild deer and boar in Japan was investigated. STEC O157 strains were isolated from 1.9% (9/474) of the wild deer and 0.7% (3/426) of the wild boar examined. Pulsed-field gel electrophoresis (PFGE) analysis classified the wild deer and boar strains into four and three PFGE patterns, respectively. The PFGE pattern of one wild boar strain was similar to that of a cattle strain that had been isolated from a farm in the same area the wild boar was caught, suggesting that a STEC O157 strain may have been transmitted between wild boar and cattle. Clade analysis indicated that, although most of the strains were classified in clade 12, two strains were classified in clade 7. Whole-genome sequence (WGS) analysis indicated that all the strains carried mdfA, a drug resistance gene for macrolide antibiotics, and also pathogenicity-related genes similar to those in the Sakai strain. In conclusion, our study emphasized the importance of food hygiene in processing meat from Japanese wild animals for human consumption.
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http://dx.doi.org/10.1292/jvms.21-0454DOI Listing
October 2021

Development of alveolar and airway cells from human iPS cells: toward SARS-CoV-2 research and drug toxicity testing.

J Toxicol Sci 2021 ;46(9):425-435

Division of Pharmacology, National Institute of Health Sciences (NIHS).

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease 2019 (COVID-19). SARS-CoV-2 enters host cells by binding with the receptor angiotensin-converting enzyme 2 (ACE2). While ACE2 is expressed in multiple cell types, it has been implicated in the clinical progression of COVID-19 as an entry point for SARS-CoV-2 into respiratory cells. Human respiratory cells, such as airway and alveolar epithelial type II (ATII) cells, are considered essential for COVID-19 research; however, primary human respiratory cells are difficult to obtain. In the present study, we generated ATII and club cells from human induced pluripotent stem cells (hiPSCs) for SARS-CoV-2 infection and drug testing. The differentiated cells expressed ATII markers (SFTPB, SFTPC, ABCA3, SLC34A2) or club cell markers (SCGB1A1 and SCGB3A2). Differentiated cells, which express ACE2 and TMPRSS2, were infected with SARS-CoV-2. Remdesivir treatment decreased intracellular SARS-CoV-2 viral replication and, furthermore, treatment with bleomycin showed cytotoxicity in a concentration-dependent manner. These data suggest that hiPSC-derived AT2 and club cells provide a useful in vitro model for drug development.
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http://dx.doi.org/10.2131/jts.46.425DOI Listing
September 2021

Long-Term Grow-Out Affects Colonization Fitness in Coincidence With Altered Microbiota and Lipid Composition in the Cecum of Laying Hens.

Front Vet Sci 2021 18;8:675570. Epub 2021 Jun 18.

Department of Animal Hygiene, Tokyo University of Agriculture, Atsugi City, Japan.

is one of the leading causes of gastrointestinal illness worldwide and is mainly transmitted from chicken through the food chain. Previous studies have provided increasing evidence that this pathogen can colonize and replicate in broiler chicken during its breeding; however, its temporal kinetics in laying hen are poorly understood. Considering the possible interaction between and gut microbiota, the current study was conducted to address the temporal dynamics of in the cecum of laying hen over 40 weeks, with possible alteration of the gut microbiota and fatty acid (FA) components. Following oral infection with 81-176, inocula were stably recovered from ceca for up to 8 weeks post-infection (.). From 16 weeks ., most birds became negative for and remained negative up to 40 weeks . 16S rRNA gene sequencing analyses revealed that most of the altered relative rRNA gene abundances occurred in the order , in which increased relative rRNA gene abundances were observed at >16 weeks . in the families , and . Lipidome analyses revealed increased levels of sterols associated with bile acid metabolisms in the cecum at 16 and/or 24 weeks . compared with those detected at 8 weeks ., suggesting that altered microbiota and bile acid metabolism might underlie the decreased colonization fitness of in the gut of laying hens.
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http://dx.doi.org/10.3389/fvets.2021.675570DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8249580PMC
June 2021

Effect of the Photocatalyst under Visible Light Irradiation in SARS-CoV-2 Stability on an Abiotic Surface.

Biocontrol Sci 2021 ;26(2):119-125

Division of Biomedical Food Research, National Institute of Health Sciences.

There is a worldwide attempt to develop prevention strategies against SARS-CoV-2 transmission. Here we examined the effectiveness of tungsten trioxide (WO)-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 logTCID /mL to 3.05 ±. 25 logTCID/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.
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http://dx.doi.org/10.4265/bio.26.119DOI Listing
June 2021

Development of ciclesonide analogues that block SARS-CoV-2 RNA replication.

Bioorg Med Chem Lett 2021 07 20;43:128052. Epub 2021 Apr 20.

Division of Organic Chemistry, National Institute of Health Sciences, 3-25-26, Tonomachi, Kawasaki, Kanagawa 210-9501, Japan; Graduate School of Medical Life Science, Yokohama City University, 1-7-29, Yokohama, Kanagawa 230-0045, Japan. Electronic address:

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.
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http://dx.doi.org/10.1016/j.bmcl.2021.128052DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8055493PMC
July 2021

Characteristic Distribution of Ciguatoxins in the Edible Parts of a Grouper, .

Toxins (Basel) 2021 03 17;13(3). Epub 2021 Mar 17.

Tama Laboratory, Japan Food Research Laboratories, 6-11-10 Nagayama, Tama, Tokyo 206-0025, Japan.

Ciguatera fish poisoning (CFP) is one of the most frequently encountered seafood poisoning syndromes; it is caused by the consumption of marine finfish contaminated with ciguatoxins (CTXs). The majority of CFP cases result from eating fish flesh, but a traditional belief exists among people that the head and viscera are more toxic and should be avoided. Unlike the viscera, scientific data to support the legendary high toxicity of the head is scarce. We prepared tissue samples from the fillet, head, and eyes taken from five yellow-edged lyretail () individuals sourced from Okinawa, Japan, and analyzed the CTXs by LC-MS/MS. Three CTXs, namely, CTX1B, 52--54-deoxyCTX1B, and 54-deoxyCTX1B, were confirmed in similar proportions. The toxins were distributed nearly evenly in the flesh, prepared separately from the fillet and head. Within the same individual specimen, the flesh in the fillet and the flesh from the head, tested separately, had the same level and composition of toxins. We, therefore, conclude that flesh samples for LC-MS/MS analysis can be taken from any part of the body. However, the tissue surrounding the eyeball displayed CTX levels two to four times higher than those of the flesh. The present study is the first to provide scientific data demonstrating the high toxicity of the eyes.
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http://dx.doi.org/10.3390/toxins13030218DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8002984PMC
March 2021

[Detection of Ciguatoxins from Fish Introduced into a Wholesale Market in Japan].

Shokuhin Eiseigaku Zasshi 2021 ;62(1):8-13

National Institute of Health Sciences.

Ciguatera fish poisoning (CFP), one of the most frequently occurring seafood poisonings due to marine finfish consumption, mainly affects the tropical and subtropical Indo-Pacific region and the Caribbean Sea. The principal class of toxins, ciguatoxins (CTXs) from the Pacific, includes more than 20 derivatives and are classified into two groups, CTX1B and CTX3C congeners, based on their skeletal structures. As part of risk management of CFP by the Japanese government, the import of certain species of fish into Japan is prohibited. Additionally, local governments recommend rejecting certain fish species caught in Japan. In this study, we used LC-MS/MS to analyze CTXs from 18 fish specimens belonging to 7 species that had been brought to a wholesale market but were disapproved for sale because of their potential danger of CFP. CTXs were detected in four specimens of Lutjanus bohar and one specimen of Variola louti. It was estimated that the two most poisonous specimens (no. 5: 0.348 μg/kg, no. 8: 0.362 μg/kg) had a toxicity of 0.05 MU/g. Consumption of 200 g of flesh from these fish could cause CFP. Thus, the guidance of the local government to disallow the sale of these fish species in the market contributed to the prevention of CFP.Only CTX1B congeners were detected in L. bohar (specimen no. 5), which had no record of the area where it captured from. It is presumed that the origin of specimen no. 5 was the same as that of the Okinawan L. bohar because the CTX compositions were similar. In two specimens (nos. 6 and 8) from Wakayama, both CTX1B and CTX3C congeners were detected. This is the first report to reveal the CTX profile in fish collected off the Honshu island in Japan.
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http://dx.doi.org/10.3358/shokueishi.62.8DOI Listing
March 2021

Phylogeny, Prevalence, and Shiga Toxin (Stx) Production of Clinical Escherichia coli O157 Clade 2 Strains Isolated in Shimane Prefecture, Japan.

Curr Microbiol 2021 Jan 23;78(1):265-273. Epub 2020 Oct 23.

Division of Biomedical Food Research, National Institute of Health Sciences, Tonomachi 3-25-26, Kawasaki-ku, Kawasaki City, Kanagawa, 210-9501, Japan.

This study investigated the genetic and pathogenic variation of the subgroups of clade 2 strains of Shiga toxin (Stx)-producing Escherichia coli (STEC) O157. A total of 111 strains of STEC O157 isolated in Shimane prefecture, Japan, were classified in clade 2 (n = 39), clade 3 (n = 16), clade 4/5 (n = 3), clade 7 (n = 14), clade 8 (n = 17), and clade 12 (n = 22) by single-nucleotide polymorphism analysis and lineage-specific polymorphism assay-6. These results showed a distinct difference from our previous study in which clade 3 strains were the most prevalent strains in three other prefectures in Japan, indicating that the clade distribution of O157 strains was different in different geographic areas in Japan. Phylogenetic analysis using insertion sequence (IS) 629 distribution data showed that clade 2 strains formed two clusters, designated 2a and 2b. Stx2 production by cluster 2b strains was significantly higher than by cluster 2a strains (P < 0.01). In addition, population genetic analysis of the clade 2 strains showed significant linkage disequilibrium in the IS629 distribution of the strains in clusters 2a and 2b (P < 0.05). The Φ values calculated using the IS629 distribution data indicated that strains in clusters 2a and 2b were genetically different (P < 0.001). Cluster 2b strains are a highly pathogenic phylogenetic group and their geographic spread may be a serious public health concern.
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http://dx.doi.org/10.1007/s00284-020-02252-4DOI Listing
January 2021

[Prevalence and Characterization of Campylobacter in Bile from Bovine Gallbladders].

Shokuhin Eiseigaku Zasshi 2020 ;61(4):126-131

Division of Biomedical Food Research, National Institute of Health Sciences.

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.
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http://dx.doi.org/10.3358/shokueishi.61.126DOI Listing
November 2020

[Current Incidence and Contamination Sources of Ascariasis in Japan].

Shokuhin Eiseigaku Zasshi 2020 ;61(4):103-108

Laboratory of Helminthology, Department of Parasitology, National Institute of Infectious Diseases.

Ascaris lumbricoides or roundworm is one of the key soil-transmitted helminths affecting humans. A small number of infections continue to occur in Japan, suggesting plant foodstuff contamination as the source of infection. To understand the current status of ascariasis incidence and to identify potential sources of infection, we extensively surveyed the available literature and collected data from testing facilities that examined clinical samples or foodstuffs. We observed that from 2002 onwards, there was a decrease in the number of ascariasis cases reported in scientific journals. Data from a clinical testing facility indicated that the number of detected cases declined remarkably from 2009. Foodstuff testing facilities reported that 11 of 10,223 plant foodstuff specimens were contaminated with anisakid nematodes but not with Ascaris. Imported kimchi was suspected as the most probable source of ascarid nematode infection, as one Ascaris egg-positive sample was detected among 60 kimchi samples in a testing facility. Therefore, the sources of Ascaris infection are still not fully known and need to be clarified to establish preventive countermeasures to safeguard Ascaris infections that continue to occur in Japan.
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http://dx.doi.org/10.3358/shokueishi.61.103DOI Listing
November 2020

Draft Genome Sequences of Non-HS-Producing Strains of Salmonella enterica Serovars Infantis, Enteritidis, Berta, and Kiambu in Japan.

Microbiol Resour Announc 2020 Jul 23;9(30). Epub 2020 Jul 23.

Department of Microbiology, Tokyo University of Agriculture, Tokyo, Japan.

We report the draft genome sequences of six strains of serovars Berta, Enteritidis, Infantis, and Kiambu, isolated from humans or chicken meats in Osaka, Japan, that were negative for hydrogen sulfide production. Their genome sizes ranged from 4,460,389 to 4,933,483 bp, with 3 to 9 rRNAs and 64 to 73 tRNAs and with coverages of 95× to 159×.
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http://dx.doi.org/10.1128/MRA.00335-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7378023PMC
July 2020

Attributing Human Foodborne Diseases to Food Sources and Water in Japan Using Analysis of Outbreak Surveillance Data.

J Food Prot 2020 Dec;83(12):2087-2094

National Institute of Health Sciences, 3-25-26, Tonomachi, Kawasaki-ku, Kawasaki City, Kanagawa 210-9501, Japan.

Abstract: In Japan, strategies for ensuring food safety have been developed without reliable scientific evidence on the relationship between foodborne diseases and food sources. This study aimed to provide information on the proportions of foodborne diseases caused by seven major causative pathogens (Campylobacter spp., Salmonella, enterohemorrhagic Escherichia coli [EHEC], Vibrio parahaemolyticus, Clostridium perfringens, Staphylococcus aureus, and norovirus) attributed to foods and to explore factors affecting changes in these source attribution proportions over time using analysis of outbreak surveillance data. For the calculation of the number of outbreaks attributed to each source, simple-food outbreaks were assigned to the single-food category in question, and complex-food outbreaks were classified under each category proportional to the estimated probability. During 2007 to 2018, 8,730 outbreaks of foodborne diseases caused by seven pathogens were reported, of which 6,690 (76.6%) were of unknown source. We estimated the following source attribution proportions of foodborne diseases: chicken products (80.3%, 95% uncertainty interval [UI] 80.1 to 80.4) for Campylobacter spp.; beef products (50.1%, UI 47.0 to 51.5) and vegetables (42.3%, UI 40.9 to 45.5) for EHEC; eggs (34.6%, UI 27.8 to 41.4) and vegetables (34.4%, UI 27.8 to 40.8) for Salmonella; finfish (50.3%, UI 33.3 to 66.7) and shellfish (49.7%, UI 33.3 to 66.7) for V. parahaemolyticus; grains and beans (57.8%, UI 49.7 to 64.9) for S. aureus; vegetables (63.6%, UI 48.5 to 74.6), chicken products (12.7%, UI 4.6 to 21.5), and beef products (11.1%, UI 8.5 to 13.1) for C. perfringens; and shellfish (75.5%, UI 74.7 to 76.2) for norovirus. In this study, we provide the best available evidence-based information to evaluate the link between foodborne diseases and foods. Our results on source attribution for Campylobacter spp. and EHEC suggest that the strict health regulations for raw beef were reflected in the proportions of these diseases attributed to this food.

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http://dx.doi.org/10.4315/JFP-20-151DOI Listing
December 2020

Plasmid Sequences of Four Large Plasmids Carrying Antimicrobial Resistance Genes in Escherichia coli Strains Isolated from Beef Cattle in Japan.

Microbiol Resour Announc 2020 May 14;9(20). Epub 2020 May 14.

Applied Microbiology, Graduate School of Agriculture, Hokkaido University, Sapporo, Hokkaido, Japan.

is a common reservoir for antimicrobial resistance genes that can be easily transformed to possess multidrug resistance through plasmid transfer. To understand multidrug resistance plasmids, we report the plasmid sequences of four large plasmids carrying a number of genes related to antimicrobial resistance that were found in strains isolated from beef cattle.
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http://dx.doi.org/10.1128/MRA.00219-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7225533PMC
May 2020

Draft Genome Sequence of Stenotrophomonas maltophilia CRB139-1, Isolated from Poultry Meat in Japan.

Microbiol Resour Announc 2020 Mar 19;9(12). Epub 2020 Mar 19.

Division of Biomedical Food Research, National Institute of Health Sciences, Kawasaki-ku, Kawasaki, Kanagawa, Japan.

is a nosocomial pathogen that primarily causes respiratory infection in humans. This pathogen is widely distributed in the environment, including in foods. Here, we report the draft genome sequence of strain CRB139-1, isolated from poultry meat in Japan. The genome size was 4,619,918 bp at 90× coverage.
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http://dx.doi.org/10.1128/MRA.00075-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7082454PMC
March 2020

PREVALENCE OF AMONG WILD SIKA DEER () AND BOARS () IN JAPAN.

J Wildl Dis 2020 04 13;56(2):270-277. Epub 2019 Dec 13.

Department of Veterinary Medicine, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.

We examined the prevalence of , including pathogenic species such as and , among wild sika deer () and boars () captured in Japan. The prevalence of in the wild deer was 75% (207/277) and in the boars was 74% (40/54). A total of 417 isolates of nine species were isolated from the animals examined: the largest number of isolates (48%, 200/417) were biotype 1A. Pathogenic 1B/O:8 were also isolated from two deer, and serogroups 3 and 4 were isolated from two boars and a deer, respectively. The pathogenic 1B/O:8 isolates carried four virulence genes (, , , and ), and serogroups 3 and 4 isolates carried three virulence genes (, , and ). Although the 1B/O:8 and isolates were sensitive to almost all the antimicrobials tested, the two 1B/O:8 isolates were resistant to azithromycin and ampicillin, and the three isolates were resistant only to azithromycin. These findings suggested that wild deer and boars might be important reservoirs for the agent causing human yersiniosis.
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http://dx.doi.org/10.7589/2019-04-094DOI Listing
April 2020

Kuma Bamboo Grass (Sasa veitchii) Extracts Exhibit Protective Effects Against Atypical Aeromonas salmonicida Infection in Goldfish (Carassius auratus).

Biocontrol Sci 2019 ;24(3):145-154

Hououdou Co., Ltd.

Atypical Aeromonas salmonicida ( i.e. subsp. achromogenes and subsp. masoucida) are one of the major opportunistic pathogens that cause ulcer diseases in a variety of fishes, in which this pathogen has become a worldwide economic threat in sectors that handle of particular high-priced ornamental fishes like varicolored carp and goldfish due to appearance damages. Here we reported that the kuma bamboo grass (Sasa veitchii) extracts (KBGE) that contained a variety of fatty acids, exhibited antibacterial activity against nine Aeromonas strains including 5 atypical A. salmonicida strains. Experimental challenges with four atypical A. salmonicida strains revealed that supplementation with 375 to 750 μg/ml of the KBGE restored the survival of goldfish in coincidence of inhibition of both bacterial replication and superoxide dismutase (SOD) activity upon infection, compared with those of untreated control. Together, our data demonstrating the antibacterial effects of the plant extracts proposes its possible implication for prevention of Aeromonas infection in the ornamental fish.
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http://dx.doi.org/10.4265/bio.24.145DOI Listing
January 2020

Phylogenetic Diversity and Antimicrobial Resistance of Campylobacter coli from Humans and Animals in Japan.

Microbes Environ 2019 Jun 21;34(2):146-154. Epub 2019 Mar 21.

Department of Animal Hygiene, Tokyo University of Agriculture.

The phylogenetic diversity and antimicrobial resistance (AMR) of Campylobacter coli from humans and animals in Japan between 2008 and 2014 were investigated. A total of 338 foodborne campylobacterioses were reported in Osaka, and C. coli was isolated from 38 cases (11.2%). In the present study, 119 C. coli strains (42 from humans, 25 each from poultry, cattle, and swine, and 2 from wild mallard) were examined by multilocus sequence typing (MLST). MLST assigned 36 sequence types (STs), including 14 novel STs; all human strains and 91% of animal strains (70/77) were assigned to the ST-828 clonal complex. The predominant human ST was ST-860 (18/42, 43%), followed by ST-1068 (8/42, 19%); these STs were also predominant in poultry (ST-860, 9/25, 36%) and cattle (ST-1068, 18/25, 72%). ST-1562 was only predominant in swine (11/25, 44.0%). Swine strains showed the greatest resistance to erythromycin (EM; 92.0%), while EM resistance was only found in 2 out of the 42 human strains examined (4.8%). All EM-resistant swine strains (n=15) exhibited a common point mutation in the 23S rRNA sequence (A2085G), and the tetO gene was detected in 22 out of the 23 TET-resistant swine strains. A whole genome sequencing analysis of four representative swine ST-1562 strains revealed abundant AMR-associated gene clusters in their genomes, suggesting horizontal gene transfer events during host adaptation. This is the first study to demonstrate the phylogenetic diversity and AMR profiles of C. coli in Japan. The present results suggest that poultry and cattle are major reservoirs, improving our knowledge on the epidemiological and ecological traits of this pathogen.
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http://dx.doi.org/10.1264/jsme2.ME18115DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6594732PMC
June 2019

Detection of Sarcocystis spp. and Shiga toxin-producing Escherichia coli in Japanese sika deer meat using a loop-mediated isothermal amplification-lateral flow strip.

J Vet Med Sci 2019 Apr 28;81(4):586-592. Epub 2019 Feb 28.

Department of Life and Environmental Sciences, Azabu University, 1-17-71 Fuchinobe, Chuo-ku, Sagamihara, Kanagawa 252-5201, Japan.

Game meat potentially harbors a number of parasitic and bacterial pathogens that cause foodborne disease. It is thus important to monitor the prevalence of such pathogens in game meats before retail and consumption to ensure consumer safety. In particular, Sarcocystis spp. and Shiga toxin-producing Escherichia coli (STEC) have been reported to be causative agents of food poisoning associated with deer meat consumption. To examine the prevalence of these microbiological agents on-site at a slaughterhouse, the rapid, simple and sensitive detection method known as the "DNA strip" has been developed, a novel tool combining loop-mediated isothermal amplification and a lateral flow strip. This assay has achieved higher sensitivity and faster than conventional PCR and is suitable for on-site inspection.
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http://dx.doi.org/10.1292/jvms.18-0372DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6483920PMC
April 2019

Effects of temperature, pH and curing on the viability of Sarcocystis, a Japanese sika deer (Cervus Nippon centralis) parasite, and the inactivation of their diarrheal toxin.

J Vet Med Sci 2018 Aug 3;80(8):1337-1344. Epub 2018 Jul 3.

Department of Food and Life Science, Azabu University, 1-17-71 Fuchinobe, Chuo-ku, Sagamihara, Kanagawa 252-5201, Japan.

Recently, the Sarcocystis parasite in horse and deer meat has been reported to be a causative agent of acute food poisoning, inducing nausea, vomiting and diarrhea. Compared with other causative agents, such as bacteria, viruses and other parasites, in deer meat, the Sarcocystis species parasite, including its stability under various conditions, is poorly understood. In this study, we assessed the viability of Sarcocystis spp. and the activity of their diarrhea toxin (a 15-kDa protein) in deer meat under conditions of freezing, cold storage, pH change and curing. In addition, the heat tolerance was assayed using purified bradyzoites. The results showed that the species lost viability by freezing at -20, -30 and -80°C for <1 hr, heating at 70°C for 1 min, alkaline treatment (pH 10.0) for 4 days and addition of salt at 2.0% for <1 day. Immunoblot assays showed that the diarrhea toxin disappeared together with the loss of viability. However, the parasite survived cooling at 0 and 4°C and acidification (pH 3.0 and 5.0) for more than 7 days with the diarrhea toxin intact. These results provide useful information for developing practical applications for the prevention of food poisoning induced by diarrheal toxin of Sarcocystis spp. in deer meat during cooking and preservation.
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http://dx.doi.org/10.1292/jvms.18-0123DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6115274PMC
August 2018

The sensitivity of commercial kits in detecting the genes of pathogenic bacteria in venison.

J Vet Med Sci 2018 Apr 19;80(4):706-709. Epub 2018 Mar 19.

Department of Food and Life Science, Azabu University, 1-17-71 Fuchinobe, Chuo, Sagamihara, Kanagawa 252-5201, Japan.

The expansion of the wild deer population is a major problem for the Japanese farm and forestry industries because their damage to farm products and vegetation results in huge economic loss. To promote game meat consumption, hygiene inspections should be performed to detect main bacterial pathogens before products are shipped. In this study, we aimed to evaluate the ability of commercial test kits to genetically detect EHEC, Salmonella and Listeria monocytogenes in venison. Our results demonstrated that the kits for three pathogens could be useful for venison as well as other domestic meat products. Our comparative study showed that the LAMP kits were more sensitive than the RT-qPCR kits in the detection of all of these pathogens.
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http://dx.doi.org/10.1292/jvms.17-0530DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5938204PMC
April 2018

Microbiological Quality Assessment of Game Meats at Retail in Japan.

J Food Prot 2017 Dec;80(12):2119-2126

8 Department of Animal Hygiene, Kitasato University, Higashi 23-35-1, Towada, Aomori 034-8628, Japan.

In this study, we examined the prevalence of Shiga toxin-producing Escherichia coli and Salmonella spp. and the distribution of indicator bacteria in 248 samples of game meats (120 venison and 128 wild boar) retailed between November 2015 and March 2016 in Japan. No Salmonella spp. were detected in any of the samples, whereas Shiga toxin-producing Escherichia coli serotype OUT:H25 (stx, eae) was isolated from one deer meat sample, suggesting a possible source for human infection. Plate count assays indicated greater prevalence of coliforms and E. coli in wild boar meat than in venison, whereas their prevalence in processing facilities showed greater variation than in animal species. The 16S rRNA ion semiconductor sequencing analysis of 24 representative samples revealed that the abundances of Acinetobacter and Arthrobacter spp. significantly correlated with the prevalence of E. coli, and quantitative PCR analyses in combination with selective plate count assay verified these correlations. To our knowledge, this is the first report to characterize the diversity of microorganisms of game meats at retail in Japan, together with identification of dominant microbiota. Our data suggest the necessity of bottom-up hygienic assessment in areas of slaughtering and processing facilities to improve microbiological safety.
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http://dx.doi.org/10.4315/0362-028X.JFP-17-137DOI Listing
December 2017

Rapid and Accurate Diagnosis Based on Real-Time PCR Cycle Threshold Value for the Identification of Campylobacter jejuni, astA Gene-Positive Escherichia coli, and eae Gene-Positive E. coli.

Jpn J Infect Dis 2018 Jan 31;71(1):79-84. Epub 2017 Oct 31.

Shimane Prefectural Institute of Public Health and Environmental Science.

We previously developed a multiplex real-time PCR assay (Rapid Foodborne Bacterial Screening 24 ver.5, [RFBS24 ver.5]) for simultaneous detection of 24 foodborne bacterial targets. Here, to overcome the discrepancy of the results from RFBS24 ver.5 and bacterial culture methods (BC), we analyzed 246 human clinical samples from 49 gastroenteritis outbreaks using RFBS24 ver.5 and evaluated the correlation between the cycle threshold (CT) value of RFBS24 ver.5 and the BC results. The results showed that the RFBS24 ver.5 was more sensitive than BC for Campylobacter jejuni and Escherichia coli harboring astA or eae, with positive predictive values (PPV) of 45.5-87.0% and a kappa coefficient (KC) of 0.60-0.92, respectively. The CTs were significantly different between BC-positive and -negative samples (p < 0.01). All RFBS24 ver.5-positive samples were BC-positive under the lower confidence interval (CI) limit of 95% or 99% for the CT of the BC-negative samples. We set the 95% or 99% CI lower limit to the determination CT (d-CT) to discriminate for assured BC-positive results (d-CTs: 27.42-30.86), and subsequently the PPVs (94.7%-100.0%) and KCs (0.89-0.95) of the 3 targets were increased. Together, we concluded that the implication of a d-CT-based approach would be a valuable tool for rapid and accurate diagnoses using the RFBS24 ver.5 system.
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http://dx.doi.org/10.7883/yoken.JJID.2017.151DOI Listing
January 2018

A childhood-onset intestinal toxemia botulism during chemotherapy for relapsed acute leukemia.

Ann Clin Microbiol Antimicrob 2017 Sep 18;16(1):61. Epub 2017 Sep 18.

Department of Pediatrics, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan.

Background: Botulism is a potentially fatal infection characterized by progressive muscle weakness, bulbar paralysis, constipation and other autonomic dysfunctions. A recent report suggested that cancer chemotherapy might increase the risk for the intestinal toxemia botulism in both adults and children.

Case Presentation: We report a 5-year-old boy, who developed general muscle weakness, constipation, ptosis and mydriasis during the third induction therapy for relapsed acute myeloid leukemia. He had recent histories of multiple antibiotic therapy for bacteremia and intake of well water at home. Repeated bacterial cultures identified Clostridium botulinum producing botulinum neurotoxin A. Botulinum toxin A was isolated from his stools at 17, 21, and 23 days after the onset. Symptoms were self-limiting, and were fully recovered without anti-botulinum toxin globulin therapy.

Conclusion: This is the second report of a pediatric case with cancer chemotherapy-associated intestinal toxemia botulism. Our case provides further evidence that the immunocompromised status due to anti-cancer treatments increases the risk for the development of botulism at all ages in childhood.
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http://dx.doi.org/10.1186/s12941-017-0240-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5604400PMC
September 2017

Genome Sequence of Strain Adk2012 Associated with a Foodborne Botulinum Case in Tottori, Japan, in 2012.

Genome Announc 2017 Aug 24;5(34). Epub 2017 Aug 24.

Department of Bacteriology II, National Institute of Infectious Diseases, Tokyo, Japan.

We report here a draft genome sequence of Adk2012 responsible for a foodborne botulism case that occurred in Tottori, Japan, in 2012. Its genome size was 2,904,173 bp, with 46 rRNAs and 54 tRNAs, at a coverage of 14.5×.
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http://dx.doi.org/10.1128/genomeA.00872-17DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5571422PMC
August 2017

Draft Genome Sequence of CAM970 and CAM962, Associated with a Large Outbreak of Foodborne Illness in Fukuoka, Japan, in 2016.

Genome Announc 2017 Jun 15;5(24). Epub 2017 Jun 15.

Health Science Section, Fukuoka City Institute of Health and Environment, Fukuoka, Japan.

Here, we report the draft genome sequences of CAM970 and CAM962, which were associated with a large outbreak of foodborne illness originating from undercooked chicken sushi in Fukuoka, Japan, in May 2016. Their genome sizes were 1,690,901 and 1,704,736 bp, with 22 and 23 rRNAs, 9 and 9 tRNAs, and 411× and 419× coverage for CAM970 and CAM962, respectively.
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http://dx.doi.org/10.1128/genomeA.00508-17DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5473265PMC
June 2017

Draft Genome Sequence of Five Shiga Toxin-Producing Strains Isolated from Wild Deer in Japan.

Genome Announc 2017 Mar 2;5(9). Epub 2017 Mar 2.

Laboratory of Animal Hygiene, Kitasato University, Aomori, Japan.

Shiga toxin-producing (STEC) is one of the major foodborne pathogens. Having observed the wide distribution of this pathogen in wild deer, we report here the draft genome sequence of five STEC strains isolated from wild deer () in Hokkaido, Japan.
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http://dx.doi.org/10.1128/genomeA.01455-16DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5334574PMC
March 2017
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