The inhibition of microbial attachment to food is important for the prevention of cross-contamination during food processing. The effect of several chemicals that were added in an Escherichia coli growth medium on the attachment of the bacterium to lettuce was investigated. E. coli ATCC 25922, which is reportedly a useful surrogate for E. coli O157:H7 in surface attachment studies, was preincubated in a nutrient broth (NB) containing sodium chloride, potassium chloride, sodium deoxycholate, sodium linear alkylbenzene sulfonate, or sorbic acid. The bacterial cells were placed in contact with cut lettuce in a saline solution at 5℃ for 24 hours. Only the addition of NaCl in the NB influenced the attachment of E. coli, Salmonella enterica subsp. Enteritidis, and Klebsiella pneumoniae to the lettuce. The attachment of E. coli showed the largest significant increase at 2% NaCl. Changes in the attachment levels were not due to surface hydrohobicity or the motility of E. coli cells. Similar results were observed for S. enterica although the variation in the degree of attachment of the latter was quite small. These results suggested that the attachment of E. coli O157:H7 to food surfaces is influenced by the bacterial growth conditions prior to food exposure and prior to the development of the biofilm; furthermore, the environmental NaCl concentration should be controlled during food processing to prevent the cross-contamination of foods with E. coli.
The subtilisin-like serine protease gene TghSS42 was cloned from biocontrol agent Trichoderma ghanense ACCC 30153. Its coding region is 1302 bp in length, encoding 433 aa with a predicted protein molecular weight of 42.5 kDa and pI of 5.53. The accession number of cDNA sequence of TghSS42 gene is KJ740359. Furthermore, the transcription of the TghSS42 gene was all up-regulated under nine different treatments by RT-qPCR analysis, and the highest transcription level of TghSS42 approached 177.29-fold at 4 h under induction with 1% (w/v) Alternaria alternata cell walls, indicating that TghSS42 could be induced by the plant or phytopathogen. Furthermore, Escherichia coli recombinant strain BL21-TghSS42 was constructed. The recombinant protease rTghSS42, with an expected molecular weight of approximately 68.5 kDa (containing 26.0 kDa GST tag), has been successfully expressed and purified from BL21-TghSS42. The purified protease rTghSS42 activity reached a peak of 18.7 U/mL at 4 h following 1.0 mM IPTG induction. The optimal enzyme reaction temperature was 40℃ and the optimal pH was 7.0. The recombinant protease rTghSS42 exerted broad-spectrum antifungal ability against Rhizoctonia solani, Fusarium oxysporum, A. alternata, Sclerotinia sclerotiorum and Cytospora chrysosperma. The inhibition rate of mycelial growth varied between 21.2% and 50.0%.
Acanthamoeba is found in seawater, fresh water, and soil and is an opportunistic pathogen that causes a potentially blinding corneal infection known as Acanthamoeba keratitis. The anti-amoeba activity of 9 fatty acid salts (potassium butyrate (C4K), caproate (C6K), caprylate (C8K), caprate (C10K), laurate (C12K), myristate (C14K), oleate (C18:1K), linoleate (C18:2K), and linolenate (C18:3K)) was tested on Acanthamoeba castellanii ATCC 30010 (trophozoites and cysts). Fatty acid salts (350 mM and pH 10.5) were prepared by mixing fatty acids with the appropriate amount of KOH. C8K, C10K, and C12K showed growth reduction of 4 log-units (99.99% suppression) in A. castellanii upon 180 min incubation at 175 mM, whereas the pH-adjusted control solution showed no effect. After the amoeba suspension was mixed with C10K or C12K, cell membrane destruction was observed. The minimum inhibitory concentration of C10K and C12K was also determined to be 2.7 mM. Confirmation tests were conducted using contact lenses to evaluate the effectiveness of C10K and C12K as multi-purpose solutions. Experiments using increasing concentrations showed reduced numbers of living cells in C10K (5.5 mM, 10.9 mM) and in C12K (5.5 mM, 10.9 mM). These results demonstrate the inhibitory activity of C10K and C12K against A. castellanii and indicate their potential as anti-amoeba agents.
Methicillin-resistant Staphylococcus aureus (MRSA) is one of the major pathogens responsible for nosocomial infections. The presence of MRSA in a hospital is detrimental to patients and to hospital management. Thus, rapid identification of MRSA is needed. Here, we report on a prospective method to rapidly discriminate of MSSA from MRSA using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and support vector machine (SVM) analysis in 160 clinical isolates of S. aureus. The predictive model was tested using 100 S. aureus isolates (50 MSSA and 50 MRSA). The identification rates were 90.0% for MSSA and 87.5% for MRSA in a 10-fold cross-validation SVM. In blind test sets, 60 S. aureus isolates (30 MSSA and 30 MRSA) were correctly classified, with identification rates of 93.3% for MSSA and 86.7% for MRSA. The method proposed in this study using the predictive model enables detection of one colony in 5 minutes, and thus is useful at clinical sites at which rapid discrimination of MRSA from MSSA is required.
Agaricus is known to have immunostimulatory and anti-tumor effects. However, the antiviral effects of Agaricus have not yet been examined. In the present study, the antiviral effects of an extract of Agaricus brasiliensis KA21 (AE) on the H1N1 influenza virus (PR8 strain) were investigated.
The anti-influenza virus effects of AE were examined by using the plaque formation inhibition test. AE inhibited the plaque formation of PR8 in a dose-dependent manner: 98 and 50% (IC50) inhibition at 2.5 and 0.99 mg/mL, respectively. To elucidate the mechanisms of AE, the direct actions and adsorption and invasion inhibition of AE were examined, and were found to have no inhibitory effect on PR8 infection. Thus, in vitro antiviral effects may somehow inhibit PR8 after the viral invasion of cells. These results demonstrated that it is expected that AE can effectively prevent the spread of the influenza virus.
Differences in the physical properties of individual cells cannot be evaluated with conventional experimental methods that are used to study groups of cells obtained from pure cultures. To examine the differences in the thermal tolerance of individual cells that are genetically identical, a method is needed to measure the thermal energy required to kill single cells. We developed a scanning thermal microscopy (SThM) system and measured the thermal conductivity of various bacterial cells, for example, spore formeing Bacillus genus and non spore-forming bacteria such as Escherichia coli. The thermal conductivity of vegetative cells (0.61 to 0.75 W/m・K) was found to be higher than that of spores (0.29 to 0.45 W/m・K). Furthermore the newly developed method enables us to estimate the thermal energy needed to kill individual cells or spores. We believe that this method can estimate the thermal energy required to achieve the cell for sterilization by heating.
Fermented vegetables are common part of Cambodian diet. The food safety status for these foods has not been investigated. This study was conducted to evaluate the microbiological hazards that contaminated fermented vegetables. A total of 68 samples of fermented vegetables were purchased randomly from five wet markets in Phnom Penh. The conventional culture methods for microbiological analysis were used. Coliform bacteria (Escherichia coli, Cronobactersakazakii, and Enterobacter spp.), opportunistic non-Entrobacteriaceae, Enterococcus spp., Staphylococcus spp., and Listeria spp. were found in these fermented foods. The highest contamination rate of Enterococcus spp. was 34% of total fermented vegetable samples, followed by Bacillus spp. coliform bacteria and E. coli (31%, 24% and 10%, respectively). The potential foodborne pathogen, C. sakazakii, was identified in one sample. Fermented mixed vegetables showed higher contamination rate of coliform bacteria (50%) than fermented single-type vegetables (13%). The results showed that fermented vegetables sold in wet market are poor in hygiene. The stage in the processing chain where contamination occurred should be identified and basic sanitary practice should be enforced to improve the food safety of fermented vegetables in Cambodia.