Vibrio vulnificus, an opportunistic human pathogen responsible for primary septicemia, initiates pathogenesis by attachment to the intestinal epithelial cells for which the motility by the polar flagellum plays an essential role. The proteomic analysis of outer membrane proteins showed that the treatment with the 1/2 minimum inhibitory concentration (MIC) of polymyxin B (a bacterial antimicrobial peptide) led to the reduced production of flagellin (a major component of the polar flagellum). Furthermore, the bacterial motility was inhibited in the presence of 1/2 MIC of polymyxin B. V. vulnificus has six flagellin genes organized into the flaFBA and flaCDE loci. The flaA was found to be expressed higher than flaC, and its expression was significantly decreased by polymyxin B. As well as polymyxin B, the 1/2 MIC of LL-37 (a human intestinal antimicrobial peptide) reduced the expression of flaA. In addition, among four fragments of LL-37, KI-18 and FK-13 containing F17KRIVQRIKDELR29 could lead to the decreased expression of flaA. Because the motility closely relates to virulence of V. vulnificus, the findings obtained herein indicate that LL-37 may reduce the bacterial virulence through inhibition of the motility via the polar flagellum.
Eighty-seven strains of Aspergillus section Restricti were isolated from five storage rooms (50 strains) and 21 houses (37 strains) between 2014 and 2020. Eleven species were identified based on their morphological characteristics and molecular phylogeny using the rRNA internal transcribed spacer (ITS) region, calmodulin (CaM), β-tubulin (benA), and RNA polymerase II second largest subunit (RPB2) sequences. A. penicillioides, which was known to cause the deterioration of cultural assets, was isolated at high frequency (73%) from the surfaces of 11 cultural assets in the storage rooms; A. clavatophorus and A. magnivesiculatus, which are closely related to A. penicillioides, were also isolated frequently (45 and 64%, respectively). Five species [A. clavatophorus (42.8%), A. penicillioides (42.8%), A. magnivesiculatus (14.3%), A. reticulatus (28.6%), and A. vitricola (28.6%)] were isolated from dust on the carpets in seven houses. Five species [A. clavatophorus (33.3%), A. penicillioides (55.5%), A. magnivesiculatus (44.4%), A. restrictus (44.4%), and A. gracilis (11.1%)] were isolated from dust on the bedding in nine houses. Using the taxonomic system described by Sklenář et al. (2017), five species (A. clavatophorus, A. magnivesiculatus, A. hordei, A. reticulatus, and A. glabripes) previously identified as A. penicillioides were confirmed as new to Japan.
Legionella pneumophila (L. pneumophila) is responsible for most Legionnaire's disease cases diagnosed worldwide. The species includes 16 serogroups, but most Legionnaire's disease cases (85.7% in Europe, 87.0% in Japan) are caused by L. pneumophila serogroup 1. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) can be used to identify the L. pneumophila serogroup. In this study, we compared three sample preparation methods that are compatible with MALDI-TOF MS: the direct colony transfer method (DCTM), on-target extraction method (OTEM), and in-tube extraction method (ITEM). The aim was to improve the low identification rates for L. pneumophila, and establish and validate a simple, rapid and robust MALDI-TOF MS-based method for routine use in microbiological laboratories for assignment of L. pneumophila isolates to serogroups and identification of reliable peak biomarkers. Using ITEM, 100.0% (29/29) of hot spring water samples and clinical isolates were correctly identified at the species level. Augmented reference spectra correctly identified all 29 strains at the species level and 29 isolates at the serogroup level, displaying sensitivity, specificity and accuracy of 100.0% for serogroup assignment. MALDI-TOF MS is a relatively inexpensive method for assignment of L. pneumophila serogroups that can serve as a first-line tool for rapid prospective typing.
This study determined the prevalence of Staphylococcus aureus in food of animal origin, investigated its antimicrobial susceptibility profiles and antimicrobial-resistant genes encoding resistance to methicillin (mecA), penicillin (blaZ), and vancomycin (vanA). Two hundred and sixty food samples, including raw retail milk, meat, and meat products, were obtained from local retail shops in Mansoura city, Egypt. The overall prevalence of S. aureus in the total examined food samples was 32.69% (85/260). Methicillin-resistant S. aureus (MRSA) was identified in 11.15% (29/260) of the tested food samples. S. aureus indicated a high resistance to nalidixic acid, penicillin, ampicillin, cefuroxime, trimethoprim/sulfamethoxazole, and azithromycin. The multiple antibiotic resistance (MAR) rate was 89.4% of the total S. aureus isolates, and MARindex ranges from 0.05-0.64. Genotypically, mecA and blaZ genes were identified in a percentage of 34.11% and 82.35%, respectively, while no isolates harbored the vanA gene. The presence of MAR S. aureus particularly, MRSA in food samples, is of great concern and represents a possible threat to the community. Therefore, the study's findings highlight the importance of establishing vigilant food safety practices for food handlers to inhibit the transmission of S. aureus through the food chain to reduce public health risks.
The aims of this study were to determine the impact of storage practice and mold types on mold growth and aflatoxin B1 (AFB1) concentration in corn residue from local seed corn plants, the main roughage source of dairy farms in the northern region in Thailand. A total of 223 samples from 2 types of corn residue - dried and wet - were collected. Mold contamination was determined by spread plate technique, and aflatoxin B1 (AFB1) quantification was performed by a commercial enzyme-linked immunosorbent assay. Multivariate linear models were created to determine factors associated with fungal quantity and AFB1 concentration. Results showed that the presence of Cladosporium spp. in the samples was associated with a lower risk of AFB1 contamination (P<0.05). In addition, appropriate storage practices, e.g. keeping feeds under a roof and using floor canvas under feed piles, gave lower risk of mold contamination and decreasing AFB1 contamination.
The mechanism of thermal death of mold conidia has not been understood in detail. The purpose of this study is to analyze the death kinetics of heated conidia of Cladosporium sphaerospermum and to ascertain the expectant cell injury responsible for the death. The death of the dormant (resting) conidia of Cladosporium sphaerospermum was examined at temperatures of between 43 and 54℃ with the conventional colony count method. The death reaction apparently followed the first order kinetics, but the Arrhenius plot of the death rate constant demonstrated seemingly a break. The linearity at temperatures higher than that at the break was lost at lower temperatures, suggesting the involvement of an unusual mechanism in the latter temperatures. In the cell morphology, we observed with quinacrine staining the vacuole rupture at a lower temperature but not at a high temperature. Interestingly, the vacuole rupture by low-temperature heating was found to correlate with the viability loss. Furthermore, active protease originally locating in vacuoles was detected in the cytoplasm of the conidia after heated at a low temperature. The results obtained suggest the involvement of potent autophagic cell death induced by low temperature heating of C. sphaerospermum conidia.