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In this study, we aimed to investigate how microbial contamination progresses on the carcass surface during the slaughter process. Cattle carcasses were tracked during a series of slaughter processes (five steps), and carcass surfaces (four parts) and equipment (nine types) were swabbed to investigate the bacterial contamination. Results showed that the outer surface (near the rear region of the flank [Top round] and [Top sirloin butt]) had significantly higher total viable counts (TVCs) than inner surface (p < 0.01) and that TVCs gradually decreased along the process. Enterobacteriaceae (EB) counts were high on the splitting saw and in top round region, and EB was detected on the inner surface of the carcasses. Furthermore, in some carcasses, Yersinia spp., Serratia spp., and Clostridium spp. present on top round and top sirloin butt immediately after skinning and remained on the carcass surface after the final process. These bacterial groups are detrimental to beef quality as they can grow in the package during cold distribution. Our results show that the skinning process is the most prone to microbial contamination, including psychrotolerant microorganisms. Moreover, this study provides information for understanding the dynamics of microbial contamination in the cattle slaughter process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The purpose of this study was to investigate the abundance and distribution of psychrophilic microorganisms associated with spoilage in beef slaughterhouse environments after cleaning. The processing lines and equipment used in slaughtering and boning were swabbed, and the microbial count was determined using a TSA and MRS medium and Chromocult® Coliform agar incubated at 15ºC and 37ºC, respectively. As a result, the brisket saw (handle side) and trolley hook were the most heavily contaminated with microorganisms, with each having a microbial adhesion rate of 66.7%. The microbial adhesion rates of the apron and milling cutter (edge side) were 50%, respectively, and those of the foot cutter (edge and handle side), splitting saw (edge side), and knife (handle side) were 33.3%, respectively. Next, four colonies were randomly isolated from the petri dish used for the bacterial count measurement to identify the predominant microbial species of the microorganisms attached to each equipment. As a result of Sanger sequencing analysis, yeasts such as Candida zeylanoides and Rhodotorula sp. and bacteria including Pseudomonas sp. and Rhodococcus sp. were identified from the equipment used in the slaughtering line, and it was assumed that these microorganisms were of environmental origin. In contrast, only Pseudomonas sp. and Candida zeylanoides were isolated from the boning line. Despite the use of cleaning operations, this study identified some equipment was contaminated with microorganisms. Since this equipment frequently comes into direct contact with the carcass, it is critical to thoroughly remove the microorganisms through accurate cleaning to prevent the spread of microbial contamination on the carcasses., Competing Interests: The authors have declared that no competing interests exist.

One of the major foods causing norovirus gastroenteritis is bivalve shellfish, such as oysters. Depuration and relaying methods have been used to control norovirus. However, these methods may be inadequate to control norovirus gastroenteritis. The present study aimed to investigate the effectiveness of high hydrostatic pressure (HHP) treatment in controlling norovirus in shelled oysters, by evaluating the inactivating effect of HHP on murine norovirus strain 1 (MNV-1) inoculated into a buffer, oyster homogenate, and shelled oysters. First, MNV-1 was inoculated (infectivity of 4.5 log PFU/mL) into the buffer and oyster homogenate, with a pH of 6.3 and salinity (NaCl) of 1.5%, mimicking the habitats of the Pacific oyster ( Crassostrea gigas ). HHP treatment at 100, 200, 275, and 300 MPa for 2 and 5 min was conducted at an initial temperature of 0 or 5°C. The infectivity of MNV-1 in both the buffer and the oyster homogenate was lower when the initial temperature was 0°C. In the buffer, the infectivity of MNV-1 decreased to 1.8 log PFU/mL after HHP treatment (200 MPa for 5 min at 0°C), and the inactivating effect was higher in the buffer than in the oyster homogenate. MNV-1 was inoculated into shelled oysters (4.8 log PFU per oyster), and HHP treatment was done at 275, 300, and 350 MPa for 5 min at the initial temperature of 0°C. The infectivity of MNV-1 decreased to 2.8 log PFU per oyster after HHP treatment at 275 MPa for 5 min. The results indicate that the inactivating effect of HHP treatment varies, depending on the medium surrounding the viral particles. Inactivation was best in buffer, followed by oyster homogenate and shelled oysters. The data could inform the development of methods to control norovirus in oysters.

Histamine in foods with a high histidine content may be produced by bacteria with histidine decarboxylase activity. Consumption of food enriched in histamine can produce symptoms of histamine poisoning that include flushing, headache, and urticaria. The number of histamine poisoning cases in Japan has decreased with developments in food hygiene management technology. However, approximately 10 cases are still reported each year. In addition, there have been cases where histamine was detected in the end products, prompting large product recalls. To prevent and identify causes of histamine toxicity, manufacturers must identify the bacteria causing the illness. A simple method of identification is needed, since sequence-based identification is complicated to perform and the analysis takes a long time. High-Resolution Melting Analysis (HRMA) is a method that detects differences in the base sequences of PCR products manifested as varied melting temperatures of double-stranded DNA. The present study was intended to develop a rapid identification method for major histamine-producing bacteria using HRMA. Species-specific HRMA primers were designed that specifically targeted the hdcA gene of 20 Gram-negative histamine-producing bacterial strains. The designed primers were used for HRM analysis of the 20 histamine-producing bacterial strains. The strains were divided into three groups (A, B, and C) based on differences in melting temperature values obtained by T m Calling analysis program. Group A comprised terrestrial bacteria, such as Morganella, Enterobacter, and Raoultella, while Groups B and C comprised marine bacteria, such as those belonging to the genera Vibrio and Photobacterium. The melting profiles obtained in Group A by HRMA were used to identify the aforementioned terrestrial bacteria. The findings indicated that HRMA can easily identify the major gram-negative histamine-producing bacteria. A flow chart was created to identify histamine-producing bacterial species. This method enables the identification of histamine-producing bacterial species more quickly and easily than conventional sequence-based methods. Therefore, the method could be valuable for food companies to screen raw materials and products and track the source of contamination, which will in turn contribute to the prevention of histamine-food poisoning and investigation of its causes.

Dahi, chanar-misti, paneer, and borhani are traditional fermented milk products in Bangladesh. We evaluated the microbial properties of these products by analysing their bacterial and fungal microbiota using classical culture-dependent and culture-independent methods, including 16S rDNA (V4) and internal transcribed spacer (ITS) region pyrosequencing. The viable lactic acid bacteria (LAB) count was 6 and 8 log colony-forming units (CFU)/g. The yeast count was about 3 to 8 log CFU/g. The counts of aerobic gram-positive and gram-negative bacteria in paneer and borhani samples were similar to that of the LAB count. Pyrosequencing results revealed a high abundance of gram-negative bacteria (Acinetobacter, Enterobacteriaceae, and Aeromonadaceae) in chanar-misti, and of Moraxellaceae in paneer. Various bacteria and fungi, including intestinal and mastitis-related microorganisms such as Bifidobacterium, Clostridium, Veillonella, and Clavispora lusitaniae, were found in chanar-misti. These results indicate that more safety processes are necessary for the preparation and manufacture of ready-to-eat (RTE) foods., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Although lactic acid bacteria (LAB) are used widely as starter cultures in the production of fermented foods, they are also responsible for food decay and deterioration. The undesirable growth of LAB in food causes spoilage, discoloration, and slime formation. Because of these adverse effects, food companies test for the presence of LAB in production areas and processed foods and consistently monitor the behavior of these bacteria. The 3M Petrifilm LAB Count Plates have recently been launched as a time-saving and simple-to-use plate designed for detecting and quantifying LAB. This study compares the abilities of Petrifilm LAB Count Plates and the de Man Rogosa Sharpe (MRS) agar medium to determine the LAB count in a variety of foods and swab samples collected from a food production area. Bacterial strains isolated from Petrifilm LAB Count Plates were identified by 16S rDNA sequence analysis to confirm the specificity of these plates for LAB. The results showed no significant difference in bacterial counts measured by using Petrifilm LAB Count Plates and MRS medium. Furthermore, all colonies growing on Petrifilm LAB Count Plates were confirmed to be LAB, while yeast colonies also formed in MRS medium. Petrifilm LAB Count Plates eliminated the plate preparation and plate inoculation steps, and the cultures could be started as soon as a diluted food sample was available. Food companies are required to establish quality controls and perform tests to check the quality of food products; the use of Petrifilm LAB Count Plates can simplify this testing process for food companies.

Hepatitis A virus (HAV) is well known worldwide as a causative virus of acute hepatitis. In recent years, numerous cases of HAV infection caused by HAV-contaminated berries have occurred around the world. Because berries are often consumed without prior heating, reliable disinfection of the raw fruit is important in order to prevent HAV outbreaks. Previous studies have found that murine norovirus strain 1 (MNV-1) and human norovirus GII.4 were inactivated in heat-denatured lysozyme solution. In this study, we investigated whether or not heat-denatured lysozyme is effective in inactivating HAV and whether it could be an effective disinfectant for berries contaminated with HAV or MNV-1. We examined the inactivating effect of heat-denatured lysozyme on three strains of HAV and found that it reduced the infectivity of all three strains. We then immersed blueberries and mixed berries into solutions of HAV or MNV-1, and disinfected them by soaking them in 1% heat-denatured lysozyme for 1min. Consequently, the infectious HAV and MNV-1 contaminating the berries were decreased by >3.1 log units in all samples. Our results demonstrate that heat-denatured lysozyme effectively inactivates HAV and suggest that heat-denatured lysozyme may be an effective disinfectant for berry fruit, which is a potential source of HAV food poisoning., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The family Enterobacteriaceae, members of which are widely distributed in the environment, includes many important human pathogens. In this study, a rapid real-time PCR method targeting rplP, coding for L16 protein, a component of the ribosome large subunit, was developed for enumerating Enterobacteriaceae strains, and its efficiency was evaluated using naturally contaminated food products. The rplP-targeted real-time PCR amplified Enterobacteriaceae species with Ct values of 14.0-22.8, whereas the Ct values for non-Enterobacteriaceae species were >30, indicating the specificity of this method for the Enterobacteriaceae. Using a calibration curve of Ct=-3.025 (log CFU/g)+37.35, which was calculated from individual plots of the cell numbers in different concentrations of 5 Enterobacteriaceae species, the rplP-targeted real-time PCR was applied to 51 food samples. A <1log difference between the real-time PCR and culture methods was obtained in a majority of the food samples (81.8%), with good correlation (r 2 =0.8285). This study demonstrated that the rplP-targeted real-time PCR method could detect and enumerate Enterobacteriaceae species in foods rapidly and accurately, and therefore, it can be used for the microbiological risk analysis of foods., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Human norovirus infects humans through the consumption of contaminated food, contact with the excrement or vomit of an infected person, and through airborne droplets that scatter the virus through the air. Being highly infectious and highly viable in the environment, inactivation of the norovirus requires a highly effective inactivating agent. In this study, we have discovered the thermal denaturing capacity of a lysozyme with known antimicrobial activity against gram-positive bacteria, as well as its inactivating effect on murine norovirus. This study is the first report on the norovirus-inactivating effects of a thermally denatured lysozyme. We observed that lysozymes heat-treated for 40 min at 100 °C caused a 4.5 log reduction in infectivity of norovirus. Transmission electron microscope analysis showed that virus particles exposed to thermally denatured lysozymes were expanded, compared to the virus before exposure. The amino acid sequence of the lysozyme was divided into three sections and the peptides of each artificially synthesised, in order to determine the region responsible for the inactivating effect. These results suggest that thermal denaturation of the lysozyme changes the protein structure, activating the region responsible for imparting an inactivating effect against the virus.

In order to study the effect of food residues on the survival of food-borne pathogens, Salmonella Typhimurium, Staphylococcus aureus, and Listeria monocytogenes were subjected to drying conditions in the presence of small amounts of food such as carrot juice, aqueous solution of nori, milk, and soy-milk. After drying for 2 h at room temperature in the absence of food residue, cell counts of S. Typhimurium, S. aureus, and L. monocytogenes decreased from 8 to 3, 6, and 5 log cfu/dish, respectively. Five milligrams of fresh carrot, 0.05 mg dried nori, and 100 nL milk or soy milk per 10 mm φ surface were sufficient to demonstrate a protective effect on the adhered pathogens, as confirmed by atomic force microscopy. Results from this study suggest that small sediments of food, not only protein rich but also carbohydrate rich, increase the resistance of surface-adherent bacteria to desiccation, rendering sanitization processes ineffective and encouraging cross contamination., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Clostridium tyrobutyricum is a gram-positive spore-forming anaerobe that is considered as the main causative agent for late blowing in cheese due to butyric acid fermentation. In this study, multilocus variable-number of tandem repeat (VNTR) analysis (MLVA) for C. tyrobutyricum was developed to identify the source of contamination by C. tyrobutyricum spores in the cheese production environment. For each contig constructed from the results of a whole genome draft sequence of C. tyrobutyricum JCM11008(T) based on next-generation sequencing, VNTR loci that were effective for typing were searched using the Tandem Repeat Finder program. Five VNTR loci were amplified by polymerase chain reaction (PCR) to determine their number of repeats by sequencing, and MLVA was conducted. 25 strains of C. tyrobutyricum isolated from the environment, raw milk, and silage were classified into 18 MLVA types (DI=0.963). Of the C. tyrobutyricum strains isolated from raw milk, natural cheese, and blown processed cheese, strains with identical MLVA type were detected, which suggested that these strains might have shifted from natural cheese to blown processed cheese. MLVA could be an effective tool for monitoring contamination of natural cheese with C. tyrobutyricum in the processed cheese production environment because of its high discriminability, thereby allowing the analyst to trace the source of contamination., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Listeria monocytogenes is the causative bacteria of listeriosis, which has a higher mortality rate than that of other causes of food poisoning. Listeria spp., of which L. monocytogenes is a member, have been isolated from food and manufacturing environments. Several methods have been published for identifying Listeria spp.; however, many of the methods cannot identify newly categorized Listeria spp. Additionally, they are often not suitable for the food industry, owing to their complexity, cost, or time consumption. Recently, high-resolution melting analysis (HRMA), which exploits DNA-sequence differences, has received attention as a simple and quick genomic typing method. In the present study, a new method for the simple, rapid, and low-cost identification of Listeria spp. has been presented using the genes rarA and ldh as targets for HRMA. DNA sequences of 9 Listeria species were first compared, and polymorphisms were identified for each species for primer design. Species specificity of each HRM curve pattern was estimated using type strains of all the species. Among the 9 species, 7 were identified by HRMA using rarA gene, including 3 new species. The remaining 2 species were identified by HRMA of ldh gene. The newly developed HRMA method was then used to assess Listeria isolates from the food industry, and the method efficiency was compared to that of identification by 16S rDNA sequence analysis. The 2 methods were in coherence for 92.6% of the samples, demonstrating the high accuracy of HRMA. The time required for identifying Listeria spp. was substantially low, and the process was considerably simplified, providing a useful and precise method for processing multiple samples per day. Our newly developed method for identifying Listeria spp. is highly valuable; its use is not limited to the food industry, and it can be used for the isolates from the natural environment.

Background: The bacterial flora of two Japanese traditional fermented fish products, aji-narezushi (salted and long-fermented horse mackerel (Trachurus japonicas) with rice) and iwashi-nukazuke (salted and long-fermented sardine (Sardinops melanostica) with rice bran), was analysed using non-culture-based polymerase chain reaction (PCR) denaturing gradient gel electrophoresis (DGGE) and culture-based PCR single-strand conformation polymorphism (SSCP) methods., Results: Viable plate counts in aji-narezushi and iwashi-nukazuke were about 6.3-6.6 and 5.7-6.9 log colony-forming units g(-1) respectively. In the PCR-DGGE analysis, Lactobacillus acidipiscis was detected as the predominant bacterium in two of three aji-narezushi samples, while Lactobacillus versmoldensis was predominant in the third sample. By the PCR-SSCP method, Lb. acidipiscis and Lactobacillus plantarum were isolated as the predominant bacteria, while Lb. versmoldensis was not detected. The predominant bacterium in two of three iwashi-nukazuke samples was Tetragenococcus muriaticus, while Tetragenococcus halophilus was predominant in the third sample., Conclusion: The results suggest that the detection of some predominant lactic acid bacteria species in fermented fish by cultivation methods is difficult., (Copyright (c) 2010 Society of Chemical Industry.)

Examination of Listeria monocytogenes prevalence among ready-to-eat foods in Japan revealed frequent (5.7 to 12.1%) contamination of minced tuna and fish roe products, and the isolates had the same virulence levels as clinical isolates in terms of invasion efficiency and infectivity in cell cultures and a murine infection model, respectively. Premature stop codons in inlA were infrequent (1 out of 39 isolates). Cell numbers of L. monocytogenes in minced tuna and salmon roe increased rapidly under inappropriate storage temperatures (from a most probable number [MPN] of 10(0) to 10(1)/g to an MPN of 10(3) to 10(4)/g over the course of 2 days at 10 degrees C). Thus, regulatory guidelines are needed for acceptable levels of L. monocytogenes in these foods.

Rapid enumeration of Escherichia coli strains by quantitative real-time PCR targeting the uidA gene was developed and confirmed for minced beef, tuna and raw oyster. Higher sensitivity (1 CFU/g of E. coli in all three food samples) was obtained by incubating for 7 h in TSB. Colony-directed E. coli specific TaqMan PCR assay could effectively distinguish colonies grown on various selective media within 1.5-h. Inspection of E. coli in food testing laboratories is important, and our rapid E. coli detection strategy will contribute to quality control in food industries.

The TaqMan assay, a quantitative real-time polymerase chain reaction (PCR), was developed to target the ToxR gene (toxR) of Vibrio vulnificus. The toxR of V. vulnificus was cloned and sequenced. Based on these results, we designed specific primers and a probe for use in the quantitative PCR assay. Twenty-nine strains of V. vulnificus that were obtained from various sources produced a single PCR product. The amount of final amplification product and threshold cycle number were the same among the strains. We used the method to detect V. vulnificus in seawater and oyster samples. We developed standard curves to quantitate V. vulnificus numbers using the PCR from seawater and oyster samples. The standard curves were not different from that of the pure culture of V. vulnificus. We found the assay was very sensitive detecting as few as 10 microbes per milliliter of seawater and oyster homogenate. Moreover, we evaluated the TaqMan assay to detect V. vulnificus in seawater samples. The numbers of V. vulnificus counted by the TaqMan assay were similar to those by a culture method in almost samples. The TaqMan assay was performed within 2 h compared to days using the culture method. The results indicate the TaqMan assay method used in this study was rapid, effective and quantitative for monitoring V. vulnificus contamination in seawater and seafoods such as oysters.

The use of molecular tools for early and rapid detection of gram-negative histamine-producing bacteria is important for preventing the accumulation of histamine in fish products. To date, no molecular detection or identification system for gram-negative histamine-producing bacteria has been developed. A molecular method that allows the rapid detection of gram-negative histamine producers by PCR and simultaneous differentiation by single-strand conformation polymorphism (SSCP) analysis using the amplification product of the histidine decarboxylase genes (hdc) was developed. A collection of 37 strains of histamine-producing bacteria (8 reference strains from culture collections and 29 isolates from fish) and 470 strains of non-histamine-producing bacteria isolated from fish were tested. Histamine production of bacteria was determined by paper chromatography and confirmed by high-performance liquid chromatography. Among 37 strains of histamine-producing bacteria, all histidine-decarboxylating gram-negative bacteria produced a PCR product, except for a strain of Citrobacter braakii. In contrast, none of the non-histamine-producing strains (470 strains) produced an amplification product. Specificity of the amplification was further confirmed by sequencing the 0.7-kbp amplification product. A phylogenetic tree of the isolates constructed using newly determined sequences of partial hdc was similar to the phylogenetic tree generated from 16S ribosomal DNA sequences. Histamine accumulation occurred when PCR amplification of hdc was positive in all of fish samples tested and the presence of powerful histamine producers was confirmed by subsequent SSCP identification. The potential application of the PCR-SSCP method as a rapid monitoring tool is discussed.

Listeria monocytogenes is a bacterium responsible for food poisoning through ready-to-eat (RTE) food products. In particular, salads are RTE products that lead to many cases of listeriosis. Such concerns have made it necessary to find a method of inhibiting Listeria growth. In this study, coleslaw and egg salads were inoculated with L. monocytogenes , followed by addition of either ferulic acid or ferulic acid + glycine/sodium acetate, and were incubated at 10 °C for a maximum of 5 days. In coleslaw, the addition of 1500 ppm ferulic acid resulted in a 1.5 log CFU/g reduction in L. monocytogenes after 5 days. In egg salad, for 5 days following the addition of 3000 ppm ferulic acid + 1% glycine/sodium acetate compound, no additional L. monocytogenes growth was observed. This study demonstrates that under particular conditions, ferulic acid has anti-bacterial properties against L. monocytogenes . Our results suggest that ferulic acid could be highly useful for inhibiting the growth of L. monocytogenes in salad products. [ABSTRACT FROM AUTHOR]

Abstract: Food processing environments are suspected to be the primary source of Listeria monocytogenes contamination of foods. We investigated the survival of L. monocytogenes on stainless steel that was soiled with food components (minced tuna, ground pork, and cabbage) followed by dehydration. Stainless steel coupons were inoculated with 107 CFU/coupon and stored up to 30 days. In comparison with Staphylococcus aureus and Salmonella Typhimurium, L. monocytogenes demonstrated the highest survivability where about 3 log CFU/coupon remained at the end of the experimental period. While survival during the early storage period was markedly affected by both food-component attachment and food type, a long dehydration period diminished the effect of those. [ABSTRACT FROM AUTHOR]

Abstract: Method comparison criteria for validating novel microbiology methods are discussed for aerobic plate count of fish samples using two rapid alternative methods (dissolved oxygen and automated MPN methods) and a reference method (AOAC 966.23, pour plate method). Results revealed that the present AOAC and ISO 16140 criteria such as requiring strict consistency in pairwise comparison may lead to overly strict validation disallowing advantageous rapid alternative methods in food microbiology. Pairwise comparison was not suitable for validating alternative microbiology methods, but the linear regression analysis followed by checking the fit to an ideal regression line was appropriate and easy-to-understand for validating the tested rapid alternative microbiology methods. Further discussion on an international level would benefit the development and use of rapid alternative methods. [Copyright &y& Elsevier]

The Rubing milk cake from Yunnan and the Yan-cai vegetable pickles from Guangdong are traditional spontaneously fermented foods in China. We evaluated the microbial properties of these products with the analysis of their bacterial and fungal microbiota using classical culture-dependent and culture-independent methods, including a 16S rDNA gene (V4) and an internal transcribed spacer (ITS) region pyrosequencing method with MiSeq system. The viable lactic acid bacteria (LAB) count was 8 and 6 log colony-forming units (CFU)/g in Rubing and Yan-cai samples, respectively. The yeast count was approximately 100–1000 times less than the LAB count in most samples, except one Yan-cai sample. In addition, the gram-negative rod count in half of the samples was similar to the LAB count. Pyrosequencing results revealed the high abundance (10%–20%) of gram-negative Pseudomonas spp. and Enterobacteriaceae in these samples. These results suggest that some of these traditional foods are undesirable as ready-to-eat (RTE) foods, even when these are typical lactic acid fermented foods. [ABSTRACT FROM AUTHOR]

A biomarker-based PCR amplification method has been proved to be an effective tool for rapid and accurate detection of Listeria contamination in food factories. However, the current biomarkers (such as iap , hly genes) used for the detection of Listeria have a problem of detection efficiency for some Listeria species, including 7 recently recognized species. Therefore, in this study, a new comprehensive biomarker was developed for the effective detection of all Listeria species. The study employed a novel in-silico scheme to explore and characterize the alternative biomarkers. Biomarker BE-LisAll was identified against 34 Listeria and over 2700 other bacterial complete genomes by in-silico scheme. Specificity of biomarker BE-LisAll was then evaluated with 17 different Listeria species and 58 non- Listeria bacteria isolates. The result showed 100% specificity to Listeria species, and the biomarker could differentiate Listeria species from a variety of non- Listeria bacteria. Finally, the PCR amplification with BE-LisAll was compared with the conventional culture method for detection of Listeria spp. using 60 swab samples collected from a food-processing factory to verify the biomarker's applicability. The result demonstrated 100% correspondence to the results of the conventional culture method. The BE-LisAll biomarker-based PCR amplification is presented for rapid and comprehensive detection of all Listeria species with a high degree of accuracy and sensitivity. It is hoped that commercial food processing industries are able to employ this developed biomarker as a Listeria -detection tool in their factories to prevent or reduce economic losses due to Listeria contamination. [ABSTRACT FROM AUTHOR]

Abstract: Tetragenococcus halophilus and Tetragenococcus muriaticus are moderately halophilic lactic acid bacteria, characteristically grouped as tetrads arrangement of cocci, are found in fermentation period of various salted and fermented foods, such as soy sauce and fish sauce. T. halophilus may be one of the most important contributor to the salted and fermented products. In this study, a MALDI-TOF MS method was developed for the rapid identification of T. halophilus and T. muriaticus. Before the MS analysis, the cells were washed with ethanol and treated with formic acid, then ionized with a matrix: α-cyano-4-hydroxycinnamic acid. The MS peak similarity in each of eleven stock strains of the same species was higher than 78%, and the similarity of the two different species was about 36%. The similarity between the two Tetragenococcus and other lactic acid cocci (LAC) including Tetragenococcus koreensis and Tetragenococcus solitarius, was only 24% and less. The MALDI-TOF MS results were obtained in 30 min, from colonies grown on agar plates for viable counts. The MALDI-TOF MS method is a very rapid, simple and accurate method for discrimination of T. halophilus from T. muriaticus and other LAC, and therefore will be useful for quality control in production of salted and fermented food. [Copyright &y& Elsevier]

Abstract: Listeria monocytogenes causes listeriosis in humans, mainly through the consumption of ready-to-eat foods such as cheese. Immunocompromised persons, the elderly, and pregnant women and their fetuses or newborns are at the highest risk for the infection. We examined the effects of dietary milk-casein (MC) and soy-protein (SP), and their digested compounds tryptone (TP) and phytone peptone (PP), respectively, on L. monocytogenes invasion and infection in human enterocyte-like Caco-2 cells and A/J mice. Invasion into Caco-2 cells tended to be high with TP. In A/J mice orally infected with L. monocytogenes, viable numbers in the liver and spleen showed a tendency of decreasing with the 20% SP diet compared to the 20% MC diet. SP suppressed the inflammation marker tumour necrosis factor-α in spleen tissue. Furthermore, bacteria lipopolysaccharide (LPS)-stimulated nitric oxide (NO) secretion from murine macrophage RAW 264.7 cells was suppressed by PP more than TP. These results suggest that major dietary proteins might affect infection and inflammation by L. monocytogenes. [Copyright &y& Elsevier]