This study aimed to establish, as a proof of concept, whether bacterial cellulose (BC)-derived plant cell wall models could be used to investigate foodborne bacterial pathogen attachment. Attachment of two strains each of Salmonella enterica and Listeria monocytogenes to four BC-derived plant cell wall models (namely, BC, BC-pectin [BCP], BC-xyloglucan [BCX], and BC-pectin-xyloglucan [BCPX]) was investigated. Chemical analysis indicated that the BCPX composite (31% cellulose, 45.6% pectin, 23.4% xyloglucan) had a composition typical of plant cell walls. The Salmonella strains attached in significantly (p<0.05) higher numbers (~6 log colony-forming units [CFU]/cm(2)) to the composites than the Listeria strains (~5 log CFU/cm(2)). Strain-specific differences were also apparent with one Salmonella strain, for example, attaching in significantly (p<0.05) higher numbers to the BCX composite than to the other composites. This study highlights the potential usefulness of these composites to understand attachment of foodborne bacteria to fresh produce.
Listeria monocytogenes, a foodborne pathogen that can cause listeriosis through the consumption of food contaminated with this pathogen. The ability of L. monocytogenes to survive in extreme conditions and cause food contaminations have become a major concern. Hence, routine microbiological food testing is necessary to prevent food contamination and outbreaks of foodborne illness. This review provides insight into the methods for cultural detection, enumeration, and molecular identification of L. monocytogenes in various food samples. There are a number of enrichment and plating media that can be used for the isolation of L. monocytogenes from food samples. Enrichment media such as buffered Listeria enrichment broth, Fraser broth, and University of Vermont Medium (UVM) Listeria enrichment broth are recommended by regulatory agencies such as Food and Drug Administration-bacteriological and analytical method (FDA-BAM), US Department of Agriculture-Food and Safety (USDA-FSIS), and International Organization for Standardization (ISO). Many plating media are available for the isolation of L. monocytogenes, for instance, polymyxin acriflavin lithium-chloride ceftazidime aesculin mannitol, Oxford, and other chromogenic media. Besides, reference methods like FDA-BAM, ISO 11290 method, and USDA-FSIS method are usually applied for the cultural detection or enumeration of L. monocytogenes. most probable number technique is applied for the enumeration of L. monocytogenes in the case of low level contamination. Molecular methods including polymerase chain reaction, multiplex polymerase chain reaction, real-time/quantitative polymerase chain reaction, nucleic acid sequence-based amplification, loop-mediated isothermal amplification, DNA microarray, and next generation sequencing technology for the detection and identification of L. monocytogenes are discussed in this review. Overall, molecular methods are rapid, sensitive, specific, time- and labor-saving. In future, there are chances for the development of new techniques for the detection and identification of foodborne with improved features.
To overcome the poor water solubility of curcumin, a curcumin-β-cyclodextrin (Cur-β-CD) complex was prepared as a novel photosensitizer. Fourier-transform infrared spectroscopy (FT-IR), differential scanning calorimetry (DSC), and X-ray diffraction (XRD) were used to verify the formation of Cur-β-CD. Furthermore, the ROS generation capacity and photodynamic bactericidal effect were measured to confirm this Cur-β-CD complex kept photodynamic activity of curcumin. The result showed Cur-β-CD could effectively generate ROS upon blue-light irradiation. The plate count assay demonstrated Cur-β-CD complex possess desirable photodynamic antibacterial effect against food-borne pathogens including Staphylococcus aureus, Listeria monocytogenes and Escherichia coli. The cell morphology determined by scanning electron microscope (SEM) and transmission electron microscope (TEM) showed Cur-β-CD could cause cell deformation, surface collapse and cell structure damage of the bacteria, resulting in the leakage of cytoplasmic; while agarose gel electrophoresis and SDS-PAGE further illustrated the inactivation mechanisms by Cur-β-CD involve bacterial DNA damage and protein degradation.
The aims of this study were to determine the prevalence and antimicrobial resistance of Listeria, Salmonella, and Yersinia spp. isolated from duck and goose intestinal contents. A total of 471 samples, including 291 duck and 180 goose intestinal contents, were purchased from wet markets between November 2008 and July 2010. Listeria, Salmonella, and Yersinia spp. were isolated from 58 (12.3%), 107 (22.7%), and 80 (17%) of the samples, respectively. It was concluded that Listeria ivanovii, Salmonella Thompson, and Yersinia enterocolitica were the predominant serovars among Listeria, Salmonella, and Yersinia spp., respectively. Moreover, resistance to tetracycline was common in Listeria (48.3%) and Salmonella spp. (63.6%), whereas 51.3% of the Yersinia spp. isolates were resistant to cephalothin. Therefore, continued surveillance of the prevalence of the pathogens and also of emerging antibiotic resistance is needed to render possible the recognition of foods that may represent risks and also ensure the effective treatment of listeriosis, salmonellosis, and yersiniosis.
The goal of this study was to determine inhibitory effect of palm kernel expeller (PKE) peptides of different degree of hydrolysis (DH %) against spore-forming bacteria Bacillus cereus, Bacillus circulans, Bacillus coagulans, Bacillus licheniformis, Bacillus megaterium, Bacillus pumilus, Bacillus stearothermophillus, Bacillus subtilis, Bacillus thuringiensis, Clostridium perfringens; and non-spore-forming bacteria Escherichia coli, Lisinibacillus sphaericus, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella Typhimurium and Staphylococcus aureus.
Lactic acid bacteria (LAB) can be isolated from traditional milk products. LAB that secrete substances that inhibit pathogenic bacteria and are resistant to acid, bile, and pepsin but not vancomycin may have potential in food applications.
This study investigated the removal of bacterial surface structures, particularly flagella, using sonication, and examined its effect on the attachment of Salmonella Typhimurium ATCC 14028 cells to plant cell walls. S. Typhimurium ATCC 14028 cells were subjected to sonication at 20 kHz to remove surface structures without affecting cell viability. Effective removal of flagella was determined by staining flagella of sonicated cells with Ryu's stain and enumerating the flagella remaining by direct microscopic counting. The attachment of sonicated S. Typhimurium cells to bacterial cellulose-based plant cell wall models and cut plant material (potato, apple, lettuce) was then evaluated. Varying concentrations of pectin and/or xyloglucan were used to produce a range of bacterial cellulose-based plant cell wall models. As compared to the non-sonicated controls, sonicated S. Typhimurium cells attached in significantly lower numbers (between 0.5 and 1.0 log CFU/cm2) to all surfaces except to the bacterial cellulose-only composite without pectin and xyloglucan. Since attachment of S. Typhimurium to the bacterial cellulose-only composite was not affected by sonication, this suggests that bacterial surface structures, particularly flagella, could have specific interactions with pectin and xyloglucan. This study indicates that sonication may have potential applications for reducing Salmonella attachment during the processing of fresh produce.
Screening of lactic acid bacteria (LAB) isolated from ewe colostrum led to the identification and isolation of Enterococcus faecium CM33 with interesting features like high survival rates under acidic or bile salts condition, high tolerance for the simulated gastrointestinal condition, and high adhesive potential to Caco-2 cells. According the inhibition of pathogen adhesion test results, this strain can reduce more than 50% adhesion capacity of Escherichia coli, Shigella flexneri, Klebsiella pneumoniae, Listeria monocytogenes, and Staphylococcus aureus to Caco-2 cells. Based on the antibiotic sensitivity test findings, E. faecium CM33 was susceptible to gentamycin, vancomycin, erythromycin, ampicillin, penicillin, tetracycline, and rifampicin, but resistant to chloramphenicol, clindamycin, and kanamycin. Upon assessment of the virulence determinants for E. faecium CM33, this strain was negative for all tested virulence genes. Furthermore, the genome of this strain was evaluated for the incidence of the known enterocin genes by specific PCR amplification and discovered the genes encoding enterocins A, 31, X, and Q. Based on this study findings, the strain E. faecium CM33 can be considered as a valuable nutraceutical and can be introduced as a new potential probiotic.
The epidermal mucus of fish contains antimicrobial agents that act as biological defence against disease. This study aims to identify antibacterial activity and protein concentration of epidermal mucus of Barbodes everetti, a Bornean endemic freshwater fish. The epidermal mucus was extracted with 3% acetic acid, 0.85% sodium chloride and crude solvents. The mucus activity against eight strains of human pathogenic bacteria, including Bacillus cereus ATCC 33019, Escherichia coli O157:H7, Listeria monocytogenes ATCC 7644, Pseudomonas aeruginosa ATCC 27853, Salmonella braenderup ATCC BAA 664, Salmonella typhimurium, Staphylococcus aureus ATCC 25933, and Vibrio cholerae, were tested. The acetic acid mucus extract of B. everetti was able to inhibit five strains of bacteria and show no activity toward E. coli O157:H7, B. cereus ATCC 33019 and L. monocytogenes ATCC 7644. Moreover, the highest protein concentration was quantified in crude extract, followed by aqueous and acetic acid extracts. This study provides a preliminary knowledge on the activity of epidermal mucus of B. everetti towards five out of the eight human pathogens tested, therefore it may contain potential sources of novel antibacterial components which could be further extracted for the production of natural antibiotics towards human-related pathogenic bacteria.
Lactic acid bacteria (LAB) are non-pathogenic bacteria that have an important role in human daily life. LAB produce antimicrobial agents, such as bacteriocins, diacetyl and hydrogen peroxide which help to extend the shelf life of food products. In this study, LAB were isolated from selected seafood; threadfin salmon and grass shrimp. Antibacterial activity of LAB extracts against Salmonella typhimurium, Listeria monocytogenes, Bacillus cereus and Escherichia coli were determined using the disc diffusion method. Three strains of LAB were selected for the characterization of antibacterial agents produced by LAB. The parameters such as pH, heat, incubation period and medium, were analyzed in this experiment. Changes in environmental parameters affected the activity of antibacterial agents. The antibacterial agents produced by LAB were generally heat stable and stable in a wide range of pH levels. However, the inhibition activity of LAB was destroyed with a heat treatment of 121°C, and the antibacterial effect was reduced at a pH of 12, which occurs in most strains. The medium containing NaCl enhanced the
antibacterial activity of P1S1 and P3S3 strains against S. typhimurium, L. monocytogenes and E. coli. Moreover, the antibacterial agents exhibited the greatest inhibition activity at incubation times between 24 and 72 h. The antibacterial activity was reduced after an incubation time of 96 h. The characterization of antibacterial agents aids in the improvement of food products safety
This study was conducted to evaluate antimicrobial properties of ethanolic extracts of the leaves of Nephelium lappaceum, Curcuma longa, Cinnamomun cassia, Durio zibethinus, Vitex trifolia, Amaranthus tricolor, Syzygium samarangense and Manihot esculenta. Antibacterial properties of the extracts were studied against fifteen strains of different gram positive and gram negative pathogenic bacteria, including Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus, Vibrio para, and Escherichia coli using the agar disk diffusion method. Among the tested extracts, only Amaranthus tricolor exhibited specific inhibition of one of the tested bacteria; Bacillus cereus. Using the microdilution method, its minimum inhibitory concentration (MIC) value was determined to be 20 mg/mL.
Natural preservatives having the great antioxidant and antimicrobial activity have been utilized in the food industry for many years. In the present study, the effect of of two brands of commercial Assam green tea infusion (represented by A and B) and 0.02% BHA/BHT on microbial growth, anti-lipid oxidation and color change were investigated in cooked beef. The green tea concentration has influenced to the results. It was found that A and B at the concentration of 250 mg/mL significantly reduced the population of Staphylococcus aureus, Listeria monocytogenes, Salmonella typhimurium and E. coli in the cooked beef to an undetectable level within 2 days of storage at 4oC. A and B also exhibited higher anti-lipid oxidation activity compared to 0.02% BHA/BHT, and control. Assam green tea infusions in cooked beef significantly increased ∆ L*
value and decreased ∆ a* and ∆ b* value (p ≤ 0.05). These indicate that Assam green tea infusion might be a potential candidate as a natural preservative for beef and other types of food.
The antibacterial activity of solvent-extracted oil of noni (Morinda citrifolia L.), spinach (Spinacia oleracea L.), lady’s finger (Abelmoschus esculentus (L.) Moench), bitter gourd (Momordica charantia Linn.), and mustard (Brassica nigra L.) seed oils, and coconut (Cocos nucifera L.) oil, palm (Elaeis guineensis L.) mesocarp in hydrolyzed and unhydrolyzed form were determined in order to explore their potential usage as antibacterial agent. The hydrolysis process that was catalyzed by immobilized lipase of Rhizomucor miehei (RMIM) showed highest hydrolytic activity with 1.0 ml of added water volume except bitter gourd seed oil and palm mesocarp oil which has maximum hydrolytic activity with added water volume of 5 ml and 2.5 ml respectively. Before hydrolysis, all oil samples did not show inhibition ring zones (IRZ) on any of the tested bacteria strains (Salmonella typhimurium, Listeria monocytogenes and Escherichia coli O157:H7). Hydrolyzed lady’s finger and bitter gourd seed oil showed IRZ on all tested bacteria strains; hydrolyzed mustard seed oil on S. typhimurium and L. monocytogenes; hydrolyzed spinach seed oil and coconut oil on L. monocytogenes; hydrolyzed noni seed oil and palm mesocarp oil did not exhibit IRZ on any of the tested bacteria strains. Most of the hydrolyzed oil exhibit an inhibition activity that was different from their respective dominant fatty acids except noni seed oil and palm mesocarp oil.
Micrococcus luteus, Listeria monocytogenes, and Bacillus cereus are major food-borne pathogenic and spoilage bacteria. Emergence of antibiotic resistance and consumer demand for foods containing less of chemical preservatives led to a search for natural antimicrobials. A study aimed at characterizing, investigating the mechanism of action and regulation of biosynthesis and evaluating the biopreservative potential of pentocin from Lactobacillus pentosus CS2 was conducted. Pentocin MQ1 is a novel bacteriocin isolated from L. pentosus CS2 of coconut shake origin. The purification strategy involved adsorption-desorption of bacteriocin followed by RP-HPLC. It has a molecular weight of 2110.672 Da as determined by MALDI-TOF mass spectrometry and a molar extinction value of 298.82 M-1 cm-1. Pentocin MQ1 is not plasmid-borne and its biosynthesis is regulated by a quorum sensing mechanism. It has a broad spectrum of antibacterial activity, exhibited high chemical, thermal and pH stability but proved sensitive to proteolytic enzymes. It is potent against M. luteus, B. cereus, and L. monocytogenes at micromolar concentrations. It is quick-acting and exhibited a bactericidal mode of action against its targets. Target killing was mediated by pore formation. We report for the first time membrane permeabilization as a mechanism of action of the pentocin from the study against Gram-positive bacteria. Pentocin MQ1 is a cell wall-associated bacteriocin. Application of pentocin MQ1 improved the microbiological quality and extended the shelf life of fresh banana. This is the first report on the biopreservation of banana using bacteriocin. These findings place pentocin MQ1 as a potential biopreservative for further evaluation in food and medical applications.
Detection of enterotoxin by targeting entFM and hblA genes in Bacillus cereus BC1 strain inoculated into ready to eat food (RTF) and drink samples using polymerase chain reaction (PCR) was conducted. The B. cereus BC1 strain was confirmed as a Bacillus diarrhoeal enterotoxin (BDE) when tested by a commercially available Enzyme-linked immunosorbent assay-BDE immunoassay (ELISA-BDE immunoassay, TECRA). In the specificity study, both enterotoxin genes were detected on chromosomal DNA of B. cereus BC1 strain by showing a specific band of 1269 bp (entFM) and 874 bp (hblA), respectively. However, none of the target genes were detected for the other 15 genomic DNA bacteria (B. cereus (ATCC 11779), B. subtilis (ATCC 6633), Campylobacter jejuni (ATCC 29428), C. coli (Jabatan Kimia Malaysia, JKM), Clostridium perfringen (ATCC 13124), Enterobacter sakazaki (ATCC 51329), Escherichia coli (ATCC 43888), E. coli (ATCC 11735), Legionella pneumophila (ATCC 33152), Listeria monocytogenes (ATCC 35967), Salmonella typhi (IMR), S. enteritidis (ATCC 13076), S. typhimurium (ATCC 14028), Shigella flexeneri (ATCC 12022) and Vibrio cholerae bengal (Institute Medical Research (IMR), Malaysia) examined. The detection limit of both genes was 0.1 ng of genomic DNA. Thus, in the presence study it is evidence that the PCR analysis targeting enterotoxin of entFM and hblA genes are suitable and useful in detecting enterotoxic B. cereus in RTFs and drinks contaminated sample.
Foodborne disease has been associated with microorganisms like bacteria, fungi, viruses and parasites. Most commonly, the outbreaks take place due to the ingestion of pathogenic bacteria like Salmonella Typhi, Escherichia coli, Staphylococcus aureus, Vibrio cholera, Campylobacter jejuni, and Listeria monocytogenes. The disease usually happens as a result of toxin secretion of the microorganisms in the intestinal tract of the infected person. Usually, the level of hygiene in the food premises reflect the quality of the food item, hence restaurant or stall with poor sanitary condition is said to be the contributor to food poisoning outbreak. In Malaysia, food poisoning cases are not rare because the hot and humid climate of this country is very suitable for the growth of the foodborne bacteria. The government is also implementing strict rules to ensure workers and owners of food premises prioritize the cleanliness of their working area. Training programme for food handlers can also help them to implement hygiene as a routine in a daily basis. A lot of studies have been done to reduce foodborne diseases. The results can give information about the types of microorganisms, and other components that affect their growth. The result is crucial to determine how the spread of foodborne bacteria can be controlled safely and the outbreak can be reduced.
Listeriosis and salmonellosis are the major foodborne illnesses worldwide. Over the last decade,
increasing reports about the antibiotic resistance of Listeria monocytogenes and Salmonella from diverse sources have prompted public health concerns, especially in developing countries with over reliance or misuse of antibiotic drugs in the treatment of humans and animals. In this study, antibiotic susceptibility profiles of 58 L. monocytogenes and 12 Salmonella Enteritidis strains from vegetable farms and retail markets in Malaysia were testedby the standard disk diffusion method. Listeria monocytogenes isolates were found to exhibit 100% resistance to penicillin G. Also, high resistance patterns were observed for meropenem (70.7%) and rifampicin (41.4%). The multiple antibiotic resistance (MAR) index of L. monocytogenes isolates ranged from 0.11 to 0.56. Besides, the antibiogram results revealed that multidrugresistant (MDR) S. Enteritidis were detected and all the S. Enteritidis isolates demonstrated resistance to at least four antibiotics. Ampicillin, amoxicillin, and trimethoprim failed to inhibit all the S. Enteritidis strains. Salmonella Enteritidis isolates also displayed high resistance to nalidixic acid (75.0%), trimethoprim-sulfamethoxazole (75.0%), and chloramphenicol (66.7%). Findings in this study indicated that vegetables could be potential sources of multidrug resistance of L. monocytogenes and S. Enteritidis, which can be a serious issue and a major concern for public health. Thus, there is a great need for surveillance programs in Malaysia to continuously monitor the antibiotic resistance profiles of important pathogens.