Uniform Suspension of Heat-Killed Staphylococcus aureus for a Positive Control Used in the Monocyte-Activation Test.

Pyrogens, classified as bacterial endotoxins and non-endotoxin pyrogens (NEPs), induce fever or shock when released into the bloodstream or spinal fluid. Recently, a monocyte-activation test (MAT) involving human cell culture has been developed to detect pyrogens in injectable products. To evaluate the sensitivity of MAT, a reference standard endotoxin was used as a positive control; however, the reactivity differed between the endotoxins and NEPs, necessitating positive controls for NEPs. This study aimed to explore a preparation method for heat-killed Staphylococcus aureus (HKSA) as a positive control for NEPs in MAT. Because S. aureus forms grape-like clusters, nine types of glass filters with pore sizes of 0.5-2.7 µm were evaluated to obtain a uniform bacterial suspension. The suspension was then heat-treated to kill the bacteria, resulting in HKSA samples. Serial dilutions of HKSA were tested by MAT using peripheral blood mononuclear cells. The interleukin-6 concentrations in the culture supernatant were measured by enzyme-linked immuno-sorbent assay to assess pyrogenic activities of HKSA. The pore sizes of the glass filters affected the uniformity of HKSA, and GF/C filter was selected for HKSA preparation. Repeated filtration improved uniformity, and a uniform suspension of HKSA was obtained through double filtration using a GF/C filter. Despite the decrease in HKSA activity as filtration frequency increased, the detection limit remained consistently unchanged. This suggests that repeated filtration can adjust the activity of HKSA to a baseline level and that a uniform suspension of HKSA exhibiting low variation is suitable as a positive control in MAT.

[Detection of Salmonella and Enumeration of Hygienic Indicator Bacteria in Dried Wood Ear Mushrooms].

An outbreak of Salmonella Stanley in the United States associated with dried wood ear mushrooms imported from China prompted us to conduct serotyping of Salmonella isolated from dried wood ear mushrooms in voluntary testing, and quantitative test for Salmonella along with enumeration of hygienic indicator bacteria in positive samples in order to evaluate the risk of Salmonella outbreak from dried wood ear mushrooms. The major serovars of Salmonella isolates obtained from 20 samples were as follows: O3,10 group-London (n=3) and Weltevreden (n=5) etc, totaling 9 strains; O4 serogroup-Saintpaul (n=2), Stanley (n=1), Typhimurium (including monophasic variant; n=3), totaling 6 strains. O7 serogroup (Potsdam) and O8 serogroup (Newport) were one strain each. Qualitative and quantitative tests for Salmonella were conducted on 10 samples with remaining amounts. As a result, one sample was 220 MPN/g, six samples were<0.6 MPN/g, and three samples were negative for Salmonella per 25 g. The mean aerobic bacterial counts and coliforms in these samples were 7.8 and 6.1 log10 CFU/g, respectively. Furthermore, qualitative test for Salmonella and enumeration of hygienic indicator bacteria were conducted on dried wood ear mushroom products (33 domestic and 30 imported products) retailed in Japan. No samples showed positive for Salmonella per 25 g, and the mean aerobic bacterial counts and coliforms were approximately 2 log10 CFU/g lower than those in the 10 samples where Salmonella was isolated during voluntary testing. While no Salmonella was detected in domestically retailed wood ear mushrooms products, the serovars associated with foodborne diseases were isolated from voluntary testing samples. It indicates that potential for consumption of Salmonella contaminated wood ear mushrooms, which is at risk of causing food poisoning.

Atypical diarrhoeagenic Escherichia coli in milk related to a large foodborne outbreak.

A foodborne outbreak related to milk cartons served in school lunches occurred in June 2021, which involved more than 1,800 cases from 25 schools. The major symptoms were abdominal pain, diarrhoea, vomiting, and fever. Although major foodborne toxins and pathogens were not detected, a specific Escherichia coli strain, serotype OUT (OgGp9):H18, was predominantly isolated from milk samples related to the outbreak and most patients tested. The strains from milk and patient stool samples were identified as the same clone by core genome multilocus sequence typing and single-nucleotide polymorphism analysis. The strain was detected in milk samples served for two days related to the foodborne outbreak at a rate of 69.6% and levels of less than ten most probable number/100 mL but not on days unrelated to the outbreak. The acid tolerance of the strain for survival in the stomach was similar to that of enterohaemorrhagic E. coli O157:H7, and the same inserts in the chu gene cluster in the acid fitness island were genetically revealed. The pathogenicity of the strain was not clear; however, it was indicated that the causative pathogen was atypical diarrhoeagenic E. coli OUT (OgGp9):H18.

Matrix-Assisted Laser Desorption and Ionization Time-of-Flight Mass Spectrometry Analysis for the Direct Detection of SARS-CoV-2 in Nasopharyngeal Swabs.

The most common diagnostic method used for coronavirus disease-2019 (COVID-19) is real-time reverse transcription polymerase chain reaction (PCR). However, it requires complex and labor-intensive procedures and involves excessive positive results derived from viral debris. We developed a method for the direct detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in nasopharyngeal swabs, which uses matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-ToF MS) to identify specific peptides from the SARS-CoV-2 nucleocapsid phosphoprotein (NP). SARS-CoV-2 viral particles were separated from biological molecules in nasopharyngeal swabs by an ultrafiltration cartridge. Further purification was performed by an anion exchange resin, and purified NP was digested into peptides using trypsin. The peptides from SARS-CoV-2 that were inoculated into nasopharyngeal swabs were detected by MALDI-ToF MS, and the limit of detection was 106.7 viral copies. This value equates to 107.9 viral copies per swab and is approximately equivalent to the viral load of contagious patients. Seven NP-derived peptides were selected as the target molecules for the detection of SARS-CoV-2 in clinical specimens. The method detected between two and seven NP-derived peptides in 19 nasopharyngeal swab specimens from contagious COVID-19 patients. These peptides were not detected in four specimens in which SARS-CoV-2 RNA was not detected by PCR. Mutated NP-derived peptides were found in some specimens, and their patterns of amino acid replacement were estimated by accurate mass. Our results provide evidence that the developed MALDI-ToF MS-based method in a combination of straightforward purification steps and a rapid detection step directly detect SARS-CoV-2-specific peptides in nasopharyngeal swabs and can be a reliable high-throughput diagnostic method for COVID-19.

Development of a Novel Real-Time Polymerase Chain Reaction Assay to Detect Escherichia albertii in Chicken Meat.

Escherichia albertii is an emerging enteropathogen. Several foodborne outbreaks of E. albertii have been reported in Japan; however, foods associated with most outbreaks remain unidentified. Therefore, polymerase chain reaction (PCR) assays detecting E. albertii specifically and sensitively are required. Primers and probe for real-time PCR assays targeting E. albertii-specific gene (EA-rtPCR) was designed. With 74 strains, including 43 E. albertii strains and several of its close relatives, EA-rtPCR specifically amplified E. albertii; therefore, the sensitivity of EA-rtPCR was then evaluated. The detection limits were 2.8 and 2.0-3.2 log colony-forming unit (CFU)/mL for E. albertii culture and enriched chicken culture inoculated with the pathogen, respectively. In addition, E. albertii was detected from 25 g of chicken meat inoculated with 0.1 log CFU of the pathogen by EA-rtPCR. The detection of E. albertii from chicken meat by EA-rtPCR was also evaluated by comparing with the nested-PCR assay, and 28 retail chicken meat and 193 dissected body parts from 21 chicken carcass were tested. One and three chicken meat were positive in the nested-PCR assay and EA-rtPCR, respectively. Fourteen carcasses had at least one body part that was positive for EA-rtPCR, and 36 and 48 samples were positive for the nested-PCR assay and EA-rtPCR, respectively. A total of 37 strains of E. albertii were isolated from seven PCR-positive samples obtained from six chicken carcass. All E. albertii isolates harbored eae gene, and were classified as E. albertii O-genotype (EAOg)3 or EAOg4 by EAO-genotyping. The EA-rtPCR developed in this study has potential to improve E. albertii detection in food and advance research on E. albertii infection.

Isolation and Characterization of Antimicrobial-Resistant Escherichia coli from Retail Meats from Roadside Butcheries in Uganda.

Retail meats are one of the main routes for spreading antimicrobial-resistant bacteria (ARB) from livestock to humans through the food chain. In African countries, retail meats are often sold at roadside butcheries without chilling or refrigeration. Retail meats in those butcheries are suspected to be contaminated by ARB, but it was not clear. In this study, we tested for the presence of antimicrobial-resistant Escherichia coli from retail meats (n = 64) from roadside butcheries in Kampala, Uganda. The meat surfaces were swabbed and inoculated on PetriFilm SEC agar to isolate E. coli. We successfully isolated E. coli from 90.6% of these retail meat samples. We identified the phylogenetic type, antimicrobial susceptibility, and antimicrobial resistance genes prevalence between retail meat isolates (n = 89). Phylogenetic type B1 was identified from 70.8% of the retail meat isolates, suggesting that the isolates originated primarily from fecal contamination during meat processing. Tetracycline (TET)-resistant isolates with tetA and/or tetB gene(s) were the most frequently detected (28.1%), followed by ampicillin (AMP) resistance genes with blaTEM (15.7%,) and sulfamethoxazole-trimethoprim (SXT) resistance genes with sul2 (15.7%). No extended-spectrum beta-lactamase-producing isolates were detected. A conjugation assay showed that resistance to AMP, TET, and SXT could be simultaneously transferred to recipients. These findings suggest that antimicrobial-resistant E. coli can easily be transferred from farms to tables from retail meats obtained from roadside butcheries.

Equine herpesvirus type 1 ORF51 encoding UL11 as an essential gene for replication in cultured cells.

Equine herpesvirus type 1 (EHV-1) UL11 is a 74-amino-acid tegument protein encoded by ORF51 of the EHV-1 genome. EHV-1 UL11 was previously reported by other researchers using the RacL22 and RacH strains to be nonessential for viral replication in cultured cells. Here, we constructed UL11 mutant viruses including a UL11 null mutant and three C-terminal truncated mutants, for further characterization of EHV-1 UL11 using bacterial artificial chromosome (BAC) technology based on the neuropathogenic strain Ab4p. EHV-1 Ab4p UL11 was localized to juxtanuclear and Golgi regions as reported by other researchers. We found that no progeny viruses were produced by transfection of fetal equine kidney cells and rabbit kidney (RK-13) cells with the UL11 null mutant and truncation mutant BAC DNAs. However, mutant viruses were generated after transfection of RK13-UL11 cells constitutively expressing EHV-1 UL11 with the mutant BAC DNAs. In conclusion, UL11 of EHV-1 Ab4p is essential for replication in cultured cells.

Shigella IpgB1 promotes bacterial entry through the ELMO-Dock180 machinery.

Shigella use a special mechanism to invade epithelial cells called 'the trigger mechanism of entry', which allows epithelial cells to trap several bacteria simultaneously. On contact, Shigella deliver effectors into epithelial cells through the type III secretion system. Here, we show that one of the effectors, IpgB1, has a pivotal role in producing membrane ruffles by exploiting the RhoG-ELMO-Dock180 pathway to stimulate Rac1 activity. Using pulldown assays, we identified engulfment and cell motility (ELMO) protein as the IpgB1 binding partner. IpgB1 colocalized with ELMO and Dock180 in membrane ruffles induced by Shigella. Shigella invasiveness and IpgB1-induced ruffles were less in ELMO- and Dock180-knockdown cells compared with wild-type cells. Membrane association of ELMO-Dock180 with ruffles were promoted when cells expressed an IpgB1-ELMO chimera, establishing that IpgB1 mimics the role of RhoG in producing membrane ruffles. Taken together, our findings show that IpgB1 mimicry is the key to invasion by Shigella.

Complete genome sequence of Chlamydia psittaci NRM_5 strain isolated from the fecal samples of a wild Indian ring-necked parakeet (Psittacula krameri manillensis) in Japan.

We report here the whole-genome sequence of the Chlamydia psittaci NRM_5 strain isolated from the fecal samples of wild Indian ring-necked parakeet (Psittacula krameri manillensis) in Japan. The sequence type is ST35, which is known to be associated with pigeons and doves, indicating the potential for transmission among bird species.

Antimicrobial resistance profiles of Campylobacter jejuni and Salmonella spp. isolated from enteritis patients in Japan.

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.

Diversity of Chlamydiales detected in pet birds privately kept in individual homes in Japan.

Chlamydia-related bacteria of the Chlamydiales order have recently been described as emerging pathogens that cause pneumonia and abortion in animals and humans. We investigated the presence of Chlamydiales using real-time polymerase chain reaction (PCR) by targeting the 16S rRNA gene of a broad range of Chlamydiales in 827 fecal samples from pet birds kept in individual homes in Japan. Of the 827 samples, 493 (59.6%) tested positive for the Chlamydiales 16S rRNA gene in the real-time PCR assay. We determined the nucleic acid sequences of PCR products from 17 Chlamydiales strains. A homology search and phylogenetic analysis using these sequences confirmed that the detected Chlamydiales included C. pecorum and a broad range of Chlamydia-related bacteria. To the best of our knowledge, this is the first study to detect a wide range of Chlamydia-related bacteria in birds.

Unique genomic sequences in a novel Mycobacterium avium subsp. hominissuis lineage enable fine scale transmission route tracing during pig movement.

Mycobacterium avium subsp. hominissuis (MAH) is one of the most prevalent mycobacteria causing non-tuberculous mycobacterial disease in humans and animals. Of note, MAH is a major cause of mycobacterial granulomatous mesenteric lymphadenitis outbreaks in pig populations. To determine the precise source of infection of MAH in a pig farm and to clarify the epidemiological relationship among pig, human and environmental MAH lineages, we collected 50 MAH isolates from pigs reared in Japan and determined draft genome sequences of 30 isolates. A variable number of tandem repeat analysis revealed that most pig MAH isolates in Japan were closely related to North American, European and Russian human isolates but not to those from East Asian human and their residential environments. Historical recombination analysis revealed that most pig isolates could be classified into SC2/4 and SC3, which contain MAH isolated from pig, European human and environmental isolates. Half of the isolates in SC2/4 had many recombination events with MAH lineages isolated from humans in East Asia. To our surprise, four isolates belonged to a new lineage (SC5) in the global MAH population. Members of SC5 had few footprints of inter-lineage recombination in the genome, and carried 80 unique genes, most of which were located on lineage specific-genomic islands. Using unique genetic features, we were able to trace the putative transmission route via their host pigs. Together, we clarify the possibility of species-specificity of MAH in addition to local adaptation. Our results highlight two transmission routes of MAH, one exposure on pig farms from the environment and the other via pig movement. Moreover, our study also warns that the evolution of MAH in pigs is influenced by MAH from patients and their residential environments, even if the MAH are genetically distinct.

Detection of anti-ebolavirus antibodies in Ghanaian pigs.

Some filoviruses such as ebolaviruses and marburgviruses, cause hemorrhagic fever in humans and nonhuman primates. Pigs are suggested to play a potential role in the filovirus ecology. We investigated the seroprevalence of filovirus infection in pigs in Ghana. Using a viral glycoprotein (GP)-based enzyme-linked immunosorbent assay, we detected filovirus-specific immunoglobulin G antibodies in 5 of 139 samples. These positive sera showed specificities to four different filovirus species. Particularly, two of the positive sera reacted to GPs of two African ebolaviruses (i.e., Ebola virus and Taï Forest virus) in Western blotting. Our results suggest that these Ghanaian pigs were exposed to multiple filoviruses and emphasize the importance of continuous monitoring of filovirus infection in pig populations in West African countries.

Chlamydial species among wild birds and livestock in the foothills of Mt. Afadjato, Ghana.

The members of family Chlamydiaceae have a broad host range and cause many kinds of diseases in humans and animals. Several cases of Chlamydiaceae being detected in atypical hosts have been reported recently. Consequently, cross-species monitoring of Chlamydia in wildlife and livestock is pertinent for public health, animal hygiene and wildlife conservation. In this study, we conducted molecular surveillance of Chlamydia in wild birds and livestock around a small village in the foothills of Mt. Afadjato, Ghana where direct contact between wildlife and livestock occurs. Among 29 captured wild birds and 63 livestock, 5 sheep, 30 goats and 28 chickens, the positive ratios of Chlamydia were 24.1%, 40.0%, 43.3% and 26.9%, respectively. Chlamydia pecorum was detected in wild birds, goats, sheep and chickens. On the basis of the variable domain 2 region of ompA, several samples from different hosts showed identical sequences and were phylogenetically located to the same clusters. In addition, using ompA, C. psittaci, C. abortus and C. gallinacea were also detected in this small habitat. Further genetic and pathogenic analyses of the chlamydial distribution in this area, which represents the interface of wild and domestic animal interactions, may improve our knowledge of their transmission among different hosts.

Survival of SARS-CoV-2 and bovine coronavirus on common surfaces of living environments.

Aerosols or saliva containing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can contaminate living environments, and viruses can be indirectly transmitted. To understand the survival potential of the virus, the viral titers of bovine coronavirus (BCoV), as a model virus, and SARS-CoV-2 were measured on porous and non-porous surfaces. The amount of infectious BCoV recovered remained relatively high on non-porous substrates. However, it quickly decreased on several non-porous surfaces such as nitrile rubber. The time taken to reach the limit of detection on non-woven masks, as a porous substrate, was longer than that of non-porous substrates. On porous substrates other than non-woven masks, the amount of virus recovered quickly decreased, and then remained at a low level. Representative substrates were tested with SARS-CoV-2. The decrease in the amount of infectious virus recovered was similar to that of BCoV, although that of SARS-CoV-2 was more rapid. RNA derived from SARS-CoV-2 was also detected using real-time PCR, and it remained on surfaces much longer than infectious virus, on all substrates. Therefore, it is important to measure the viral titer to avoid the overestimation of infectious virus contamination in the environments. Our results suggest that the surface structure was not directly related to viral survivability.

Detection of Chlamydophila psittaci by using SYBR green real-time PCR.

Chlamydophila psittaci is the causative agent of human psittacosis and avian chlamydiosis. This zoonotic pathogen is frequently transmitted from infected birds to humans. Therefore proper and rapid detection of C. psittaci in birds is important to control this disease. We developed a method for detecting C. psittaci by using SYBR Green Real-time PCR based on targeting the cysteine-rich protein gene (envB) of C. psittaci. This one step procedure was highly sensitive and rapid for detection and quantification of C. psittaci from fecal samples. This assay was also able to detect other zoonotic Chlamydophila species such as C. abortus and C. felis. The assay is well suited for use as a routine detection method in veterinary medicine.

Protected but not from Contamination: Antimicrobial Resistance Profiles of Bacteria from Birds in a Ghanaian Forest Protected Area.

Resistance to antimicrobial agents is a growing concern in public health. It has been reported in wildlife from several places in the world though wild animals are not normally exposed to clinically used antimicrobial agents. Despite this, very little research has been done in Ghana to determine antimicrobial resistance in wild animals, particularly those in protected areas. In this study, the presence of colistin resistant and multidrug resistant (MDR) gram-negative bacteria in cloacal swabs of wild birds captured in a Ghanaian forest protected area were evaluated. A total of 195 isolates from 138 individual birds were obtained, identified and tested for resistance to colistin. The colistin-resistant isolates were subsequently tested for multidrug resistance to 4 other antimicrobial agents (Oxytetracycline, Streptomycin, Ampicillin and Ciprofloxacin). Colistin resistance was observed in 6.5% (9/138) of the birds and this was seen in only birds that were sampled close to the reception area of the protected area. About 50% of the colistin-resistant isolates were multidrug resistant. AMR isolates were obtained from birds that have been documented to show an insectivorous or omnivorous feeding preference. Data obtained from the study suggests that AMR and MDR occurred in wild birds from the Conservation Area and supports the claim that proximity to human impacted habitats (settlements/farmlands) increased the likelihood of carriage of AMR. Though the routes of transmission remain unclear, there is potential for spread from the wild birds to other wild/domestic animals and possibly back to humans.

Association between the blaCTX-M-14-harboring Escherichia coli Isolated from Weasels and Domestic Animals Reared on a University Campus.

Antimicrobial-resistant (AMR) bacteria affect human and animal health worldwide. Here, CTX-M-14-producing Escherichia coli isolates were isolated from Siberian weasels (Mustela sibirica) that were captured on a veterinary campus. To clarify the source of bacteria in the weasels, we examined the domestic animals reared in seven facilities on the campus. Extended-spectrum ß-lactamase (ESBL)-producing E. coli were isolated on deoxycholate hydrogen sulfide lactose agar, containing cephalexin (50 µg/mL) or cefotaxime (2 µg/mL), and were characterized with antimicrobial susceptibility testing, pulsed-field gel electrophoresis (PFGE), replicon typing, and ß-lactamase typing analyses. Next-generation sequencing of the ESBL-encoding plasmids was also performed. CTX-M-14 producers isolated from both domestic animals and weasels were classified into six clusters with seven PFGE profiles. The PFGE and antimicrobial resistance profiles were characterized by the animal facility. All CTX-M-14 plasmids belonged to the IncI1 type with a similar size (98.9-99.3 kb), except for one plasmid that was 105.5 kb in length. The unweighted pair group method with arithmetic mean (UPGMA) revealed that the CTX-M-14 plasmid in the weasel isolates might have the same origin as the CTX-M-14 plasmid in the domestic animals. Our findings shed further light on the association of antimicrobial resistance between wild and domestic animals.

Evaluating Methods for Detecting Escherichia albertii in Chicken Meat.

ABSTRACT: Escherichia albertii is an emerging foodborne pathogen. The source of the E. albertii infection in most foodborne outbreaks is unknown because E. albertii is difficult to isolate from suspected food or water. E. albertii has a broad host range among birds and can be isolated from chicken meat. In this study, PCR assay, enrichment, and isolation conditions for detecting E. albertii in chicken meat were evaluated. The growth of 47 E. albertii strains isolated in Japan between 1994 and 2018 and a type strain was evaluated in modified EC broth (mEC) and mEC supplemented with novobiocin (NmEC) and on media containing carbohydrates. The enzyme used for the nested PCR, the enrichment conditions, the most-probable-number (MPN) method, and agar media were also evaluated with chicken meat. To distinguish E. albertii from presumptive non-E. albertii bacteria, desoxycholate hydrogen sulfide lactose agar (DHL), MacConkey agar (MAC), and these agars supplemented with rhamnose and xylose (RX-DHL and RX-MAC, respectively) were used. All E. albertii strains grew in mEC and NmEC at both 36 and 42°C and did not utilize rhamnose, sucrose, or xylose. Both the first and nested PCRs with TaKaRa Ex Taq, which was 10 to 100 times more active than the other enzymes, produced positive results in enrichment culture of 25 g of chicken meat inoculated with >20 CFU of E. albertii and incubated in mEC and NmEC at 42°C for 22 ± 2 h. Thus, the first PCR was sensitive enough to detect E. albertii in chicken meat. The MPN values in mEC and NmEC were 0.5- and 2.3-fold higher than the original inoculated bacterial levels, respectively. E. albertii in chicken meat was more efficiently isolated with enrichment in NmEC (70.1 to 100%) and plating onto RX-DHL (85.4%) and RX-MAC (100%) compared with enrichment in mEC (53.5 to 83.3%) and plating onto DHL (70.1%) and MAC (92.4%). Thus, optimized conditions for the surveillance of E. albertii contamination in food and investigations of E. albertii outbreaks, including the infectious dose, were clarified.

Antimicrobial Resistance in Salmonella Isolated from Food Workers and Chicken Products in Japan.

Salmonella is an enteric bacterial pathogen that causes foodborne illness in humans. Third-generation cephalosporin (TGC) resistance in Salmonella remains a global concern. Food workers may represent a reservoir of Salmonella, thus potentially contaminating food products. Therefore, we aimed to investigate the prevalence of Salmonella in food workers and characterize the isolates by serotyping and antimicrobial susceptibility testing. Salmonella was isolated from 583 (0.079%) of 740,635 stool samples collected from food workers between January and December 2018, and then serotyped into 76 Salmonella enterica serovars and 22 untypeable Salmonella strains. High rates of antimicrobial resistance were observed for streptomycin (51.1%), tetracycline (33.1%), and kanamycin (18.4%). Although isolates were susceptible to ciprofloxacin, 12 (2.1%) strains (one S. Infantis, one S. Manhattan, two S. Bareilly, two S. Blockley, two S. Heidelberg, two S. Minnesota, one S. Goldcoast, and one untypeable Salmonella strain) were resistant to the TGC cefotaxime, all of which harbored ß-lactamase genes (blaCMY-2, blaCTX-M-15, blaCTX-M-55, and blaTEM-52B). Moreover, 1.3% (4/309) of Salmonella strains (three S. Infantis and one S. Manhattan strains) isolated from chicken products were resistant to cefotaxime and harbored blaCMY-2 or blaTEM-52B. Thus, food workers may acquire TGC-resistant Salmonella after the ingestion of contaminated chicken products and further contaminate food products.