Comparison between pre-enrichment in single- or double-strength buffered peptone water for recovery of Salmonella enterica serovar Typhimurium DT104 from acidic marinade sauces containing spices

The ability of the standard pre-enrichment procedure in buffered peptone water (BPW) to recover Salmonella Typhimurium from acidic marinade sauces containing spices was tested by inoculating marinade sauces with known numbers of an antibiotic-resistant marker strain of Salmonella Typhimurium DT104 prior to pre-enrichment. Viable numbers of salmonellae present in BPW after 24 h incubation depended on the inoculum level. If initial cell numbers were low (below 10³ cfu per 250 ml BPW) final cell concentrations were also low and, in some cases, no growth occurred. The problem was overcome by use of double-strength BPW that neutralised the acidity and allowed good recovery from otherwise inhibitory marinade sauces.     

Use of gamma irradiation for inactivation of pathogens inoculated into Kimbab,steamed rice rolled by dried laver

The effect of gamma irradiation for inactivating the pathogens inoculated into the ready-to-eat Kimbab, steamed rice rolled by dried laver, was investigated. The pathogens used were Salmonella Typhimurium, Escherichia coli, Staphylococcus aureus and Listeria ivanovii which are important for public health. The growth of four test organisms inoculated (about 10⁶–10⁷ CFU/g) into the Kimbab were sustained by an irradiation treatment during 24 h of storage regardless of the temperature at 10, 20 and 30 °C. Four pathogen inoculated into Kimbab decreased 2–3 log CFU/g by 1 kGy treatment and was not detected after 3 kGy. The D₁₀ value of pathogens inoculated into the Kimbab were 0.31–0.44 kGy among the four organisms. This study indicated that a low dose irradiation can maintain microbial safety for ready-to-eat Kimbab, steamed rice rolled by dried laver.     

In vitro antibacterial activity of Australian native herb extracts against food-related bacteria

The antibacterial activities of water, ethanol and hexane extracts of five Australian herbs (Backhousia citriodora, Anetholea anisata, Eucalyptus staigerana, Eu. olida and Prostanthera incisa) against seven food-related bacteria (Enterococcus faecalis, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella Enteritidis, Sal. Typhimurium and Staphylococcus aureus) were determined by the microtitre broth microdilution assay. The water extracts of all the herbs displayed no or low antimicrobial activity against all of the bacteria tested with the exception of S. aureus. Relatively high levels of activity (minimum inhibitory concentrations of 125–15.6 μg ml⁻¹) against this pathogen were present in water extracts from all herbs except P. incisa. The ethanol and hexane extracts of all herbs displayed some activity against a number of the bacteria tested, with no one particular herb displaying an obviously higher level or range of activity. Staphylococcus aureus proved to be the most sensitive of the bacteria tested against the solvent extracts with all extracts displaying activity ranging from 125 to 7.8 μg ml⁻¹, while E. coli and L. monocytogenes, on the other hand, proved the least sensitive with only five of 15 herb/extract combinations displaying any activity against these pathogens. The extracts of the Australian native herbs examined in this study have potential for application in foods to increase shelf-life or promote safety.     

Prevalence,genetic characterization and antimicrobial resistance of Salmonella isolated from fresh pork sausages in Porto Alegre,Brazil

A total of 336 samples of fresh pork sausage randomly obtained from supermarkets and butcher shops in Porto Alegre, Brazil, were examined for the presence of Salmonella serovars. Salmonella enterica was detected in 82 (24.4%) of the samples, with a most probable number count ranging from 0.03 MPN g^?1 to 460 MPN g^?1. Strains belonging to the most isolated S. enterica serovars (Brandenburg, Panama, Derby and Typhimurium) were further characterized by XbaI-macrorestriction, resulting in a total of 17 profiles. Resistance to tetracycline was the most prevalent among the Salmonella isolates. S. panama and S. typhimurium presented the greatest number of resistance phenotypes.     

Effects of lactic acid and hot water treatments on Salmonella Typhimurium and Listeria monocytogenes on beef

The present study is designed to determine alone and combined effects of lactic acid (LA) and hot water (HW) treatments. Hot water and different concentrations of lactic acid were evaluated for their reduction effects on Salmonella Typhimurium and Listeria monocytogenes on contaminated beef during refrigerated storage at 4 °C. The reductions were 0.05–1.19 and 0.09–1.14 log for S. Typhimurium and L. monocytogenes on day 0 respectively, while it was 0.43–1.78 and 1.69–3.84 log respectively on day 5 of storage. Results of this study suggest that LA and HW treatments can be used to reduce S. Typhimurium and L. monocytogenes which provide an additional measure of safety in production line.     

Rapid and sensitive identification of buffalo’s,cattle’s and sheep’s milk using species-specific PCR and PCR–RFLP techniques

For the rapid, specific and sensitive identification of buffalo’s, cattle’s and sheep’s milk, species-specific PCR and PCR–RFLP techniques were developed. DNA from small amount of fresh milk (100 μL) was extracted to amplify the gene encoding species-specific repeat (SSR) region and the mitochondrial DNA segment (cytochrome-b gene). PCR amplification size of the gene encoding SSR region was 603 bp in both buffalo’s and cattle’s milk, while in sheep’s milk was 374 bp. Polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) technique was used to discriminate between buffalo’s and cattle’s milk. Restriction analysis of PCR–RFLP of the mitochondrial cytochrome-b segment (359 bp) analysis showed difference between buffalo’s and cattle’s milk. Where, the fragment length (bp) generated by TaqI PCR–RFLP were 191 and 168, whereas no fragments were obtained in cattle’s milk for cytochrome-b gene (359 bp). The proposed PCR and PCR–RFLP assays rep resent a rapid and sensitive method applicable to the detection and authentication of milk species-specific.     

Sensory quality and food safety of boneless chicken breast portions thawed rapidly by submersion in hot water

Boneless chicken breast portions were thawed by submersion in hot water (60 °C) and compared to refrigerator thawing. Thawing in hot water was significantly quicker (2–8.5 min) than refrigerator thawing (10–15.5 h). Thawing time in hot water increased with an increase in meat thickness. Sensory panelists could not distinguish a difference between hot water versus refrigerator thawed and subsequently grilled chicken breast portions. A model for Salmonella growth predicts that thawing chicken breast at the slowest rate in this study (0.5 °C/min) would result in a lower increase in the Salmonella concentration than that expected for room temperature storage for 4 h.     

Effects of a partially digested whey protein concentrate on Salmonella enterica serotype Typhimurium adhesion to Caco-2 cells: Prevention of Salmonella adhesion to Caco-2 cells using whey

The effect of a partially lipase-digested whey protein concentrate (WPD) containing free fatty acids on the adherence of Salmonella enterica serovar Typhimurium to Caco-2 cells was investigated. A short concurrent exposure of the Caco-2 monolayers and WPD caused dose-dependent inhibition of bacterial adhesion. Pre-treatment of S. Typhimurium with WPD resulted in inhibition of adhesion equivalent to that with concurrent administration, whereas treatment with WPD did not significantly inhibit adhesion. WPD was not bactericidal towards S. Typhimurium at up to 50 mg ml⁻¹ for up to 1 h, but was bacteriostatic over 12 h. High concentrations of WPD (>20 mg ml⁻¹) were cytotoxic to newly-confluent cell monolayers, depending on the duration of exposure. Lactoferrin (LF), caseinoglycomacropeptide (CGMP), and gamma globulins all significantly inhibited S. Typhimurium adhesion.     

Extraction and detection of endogenous soybean DNA from fermented foods

We examined rapid and simple genomic DNA extraction method to detect material soybeans from natto (fermented soybeans) and soy sauce. Genomic DNA was extracted from natto and soy sauce by CTAB or alkaline lysis methods. Genomic DNA solutions from natto by both extraction methods were of adequate purity and yield for applying PCR, resulting that amplifications of the expected 100 bp fragment using the soybean specific primer pair were detected. When genomic DNA was extracted from soy sauce, the DNA solutions by alkaline lysis method were of a sufficient purity (A₂₆₀/A₂₈₀ = 1.5–2.0), but those by CTAB method were insufficient (A₂₆₀/A₂₈₀ = 1.2–1.7). In addition, the amplification products from the genomic DNA solutions by CTAB method were not detected. Thus, these results suggested that the desirable genomic DNA was not extracted from soy sauce by CTAB method, although it was possible to extract it by alkaline lysis method. Furthermore, when genomic DNA was extracted from natto and soy sauce using FTA^® card, which can preserve nucleic acids dried on the card just by applying a sample to it, the amplification was detected. It is found that an endogenous gene (lectin 1 gene) of material soybeans can be extracted by rapid and simple methods, and detected from fermented foods as natto and soy sauce.     

Influence of the hydrophobicity and surface roughness of mangoes and tomatoes on the adhesion of Salmonella enterica serovar Typhimurium and evaluation of cleaning procedures using surfactin

Salmonellosis is associated with the consumption of raw vegetables and fruits such as tomatoes, watermelons, alfalfa sprouts, radishes, carrots, lettuce and parsley. The influence of the fruits' roughness on bacterial adhesion was evaluated as measured using a profilometer. The adhesion of Salmonella enterica serovar Typhimurium to mango and tomato surfaces was also evaluated by measuring of the hydrophobicity of the microorganisms and the fruits surfaces. The bacteria adherent on fruit's surface was quantified by plate count and visualize by scanning electron microscopy. In addition, the efficiency of surfactin in removing S. Typhimurium from the fruits' surfaces was analyzed. The average roughness (R_a) of mango (4.54 ± 1.95 μm) was significantly different (p < 0.05) compared to tomato (2.88 ± 2.15 μm). The adhesion of the microorganisms to the fruits' surfaces, as predicted by a determination of the total energy of adhesion (ΔG), was thermodynamically unfavorable. Despite these data, the numbers of bacteria on both fruits' surfaces were similar (p > 0.05), reaching 5.95 ± 0.36 log CFU cm⁻² and 5.81 ± 0.39 log CFU cm⁻² on mango and tomato, respectively. Therefore, these results suggest that the adhesion observed in this experiment is a multifactorial process. Surfactin removed 94.3% and 92.2% of the S. Typhimurium adhered to the surfaces of the mangoes and tomatoes, respectively. Our research showed that the roughness and hydrophobicity of the fruits' surface did not affect the efficiency of each sanitation treatments on removing of S. Typhimurium. It was observed that the chlorine was more efficient treatment (p < 0.05) for tomato surface. For surface of mangoes, chorine and surfactin were better than water treatment for bacteria control.     

Hot air treatment for surface decontamination of table eggs

A hot air assay was set up for the surface decontamination of table eggs experimentally contaminated by Salmonella enterica serovar Enteritidis. A hot air apparatus was built and a treatment of two shots of 8 s at 600 °C with an interval of 30 s of cold air was chosen and applied on contaminated eggs. The S. Enteritidis load on eggshells as well as the quality traits of the egg components of 190 treated and 190 not treated eggs was investigated during 24 days of storage at 20 °C. Hot air treatment reduced the S. Enteritidis load on eggshells of up to 1.9 log. No significant changes to any of the quality traits tested were recorded. These results suggest the usefulness of hot air pasteurisation for the surface decontamination of table eggs.     

Inhibitory effects of selected plant essential oils on the growth of four pathogenic bacteria: E. coli O157:H7, Salmonella Typhimurium,Staphylococcus aureus and Listeria monocytogenes

Twenty eight essential oils were evaluated for their antibacterial properties, against four pathogenic bacteria (Escherichia coli O157:H7, Listeria monocytogenes 2812 1/2a, Salmonella Typhimurium SL 1344 and Staphylococcus aureus). Essential oils were introduced into Brain Heart Infusion agar (BHI) (15 ml) at a concentration of 0.003%, 0.006%, 0.013%, 0.025%, 0.05%, 0.1%, 0.2%, 0.4% and 0.8% (vol/vol) to determine the minimum inhibitory concentration (MIC) and the maximal tolerated concentration (MTC) for each pathogen evaluated. Results showed that the most active essential oils against bacteria tested were Corydothymus capitatus, Cinnamomum cassia, Origanum heracleoticum, Satureja montana, and Cinnamomum verum (bark). These showed a MIC ⩽ 0.05% (vol/vol) for all bacteria tested. For the MTC, with the exception of S. Typhimurium and L. monocytogenes where a MTC of 0.025% (vol/vol) was observed in presence of Cinnamomum verum and Cinnamomum cassia, respectively, a MTC ⩽ 0.013% (vol/vol) was observed for all other bacteria and the three other most active essential oils. Three oils (Satureja hortensis, Thymus vulgaris carvacroliferum, Origanum compactum) showed a MIC ⩽ 0.1% (vol/vol) for all bacteria tested. Seven oils (Thymus vulgaris thymoliferum, Thymus serpyllum, Thymus satureioides, Cymbopogon martinii, Pimenta dioica, Cinnamomum verum (leaf), Eugenia caryophyllus) showed a lower antimicrobial activity showing a MIC ⩽ 0.4% (vol/vol) against the four bacteria tested. Finally, 13 essential oils were less active showing a MIC value ⩾ 0.8% (vol/vol) against at least one bacterium.     

Efficiency of high pressure treatment on inactivation of pathogenic microorganisms and enzymes in apple,orange, apricot and sour cherry juices

The purpose of this study was to investigate the effect of high hydrostatic pressure with a mild heat treatment on Staphylococcus aureus 485, Escherichia coli O157:H7 933 and Salmonella Enteritidis FDA in apple, orange, apricot and sour cherry juices. The effectiveness of the treatment on polyphenol oxidase activity in apple juice and pectinesterase activity in orange juice were also determined. An inoculum of microorganisms was completely inactivated at 350 MPa and 40 °C in 5 min. The residual polyphenol oxidase activity in apple juice after treatment at 450 MPa and 50 °C for 60 min was obtained as 9 ± 2.2%. The residual pectinesterase activity in orange juice after treatment at 450 MPa and 50 °C for 30 min was determined as approximately 7 ± 1.6%. It compares with 12 ± 0.2% at a treatment of 40 °C and 450 MPa for 60 min. Pressure resistant isoenzymes were thought to be responsible for the final residual activity. The inactivation is irreversible and the enzyme is not reactivated upon storage. High pressure processing constitutes an effective technology to inactivate the enzymes in fruit juices. Pressures higher than 400 MPa can be combined with mild heat (<50 °C) to accelerate enzyme inactivation.     

Inactivation of foodborne pathogens in raw milk using high hydrostatic pressure

Plate counting, viability test, pulse field gel electrophoresis (PFGE) and transmission electron microscopy (TEM) were used to investigate the effect of high hydrostatic pressure (HHP) on Salmonella, Escherichia coli, Shigella and Staphyloccocus aureus in raw milk in order to determine the optimal inactivation conditions, and further understand the mechanisms of HHP on pathogens inactivation in food. The results exhibited that 300 Mpa treatment with 30 min duration at 25 °C was the optimal condition for Salmonella, E. coli, Shigella and S. aureus inactivation. Damage of the cell wall, cell membrane and cytoplasmic components by high pressure treatment can be observed in TEM micrographs. The injured cells could not be recovered, the growth rate of survivors was much lower than that of the untreated cells. PFGE showed neither corresponding DNA bands with same molecular weight nor DNA bands with same brightness could be found in the lanes between HHP treated pathogens and untreated ones. The results indicated that HHP processing can be applied to inactivate pathogens in food, the inactivation is mainly due to cell membrane damage, cell wall rupture and chromosome DNA degradation.     

Inactivation of Salmonella,E. coli and Listeria monocytogenes in phosphate-buffered saline and apple juice by ultraviolet and heat treatments

Two strains of Escherichia coli (K-12 and O157:H7), Salmonella (enteritidis and typhimurium) and Listeria monocytogenes (AS-1 and M24-1) were individually suspended in phosphate-buffered saline (PBS) and apple juice prior to exposure to UV radiation (220–300 nm) and heating at 55 °C. The calculated decimal reduction times (D value, min) varied with suspending medium and mode of inactivation. The AS-1 and M24-1 strains of L. monocytogenes were found to be most resistant to UV in PBS (0.28–0.29 min) while the AS-1 strain was most resistant in juice (1.26 min). The AS-1 strain of L. monocytogenes and E. coli O157:H7 were most heat resistant when suspended in PBS (4.41 min) and juice (4.43 min), respectively. Results obtained from this study may be used by apple juice processors in selecting appropriate organisms for UV irradiation or heat treatment lethality validations.     

Incidence of pathogenic psychrotrophs in ice creams sold in some retail outlets in Mumbai,India

With a view to determine the microbial quality of ice creams, 30 samples of commercial brands of three flavours sold in the `open' and `packaged, (cone, cup)' forms were analyzed for their total bacterial counts (TBC), yeast and mold counts (YMC), coliforms and pathogenic psychrotrophs; Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, Yersinia enterocolitica and Salmonella spp. In general in both the types of ice creams bacterial load (2.3 × 10⁴–8.5 × 10⁶ cfu/ml) was higher; particularly coliform levels were 10–100 fold higher (3.0 × 10²–5.8 × 10⁴ cfu/ml) than the safety limits prescribed by Indian Standards Institute (ISI). Staph. aureus was detected in both the types of ice creams but occurrence of B. cereus was more frequent in open samples (40%) than in packed ones (26.6%). Salmonella was not detected in any of the 30 samples tested. While 53% of the packed and 100% of the open ice creams exhibited Listeria contamination, Yersinia were detected in 33% of packed and 40% of open ones. L. monocytogenes and/or Y. enterocolitica was detected only in one of the open ice cream samples. Growth profile of Y. enterocolitica 5692 and L. monocytogenes 036 at simulated temperature abuse conditions during commercial frozen storage showed that after 10 days L. monocytogenes could grow to >1 log and 1 log cycle at 8–10°C and 2–4°C, respectively and Y. enterocolitica grew 2 log cycles at both the temperatures. Results are discussed in the context of present microbiological specifications and the need for its implementation by regulatory agencies to ward off possible health hazards arising from pathogens.     

Application of slightly acidic electrolyzed water as a potential non-thermal food sanitizer for decontamination of fresh ready-to-eat vegetables and sprouts

The sanitization efficacy of slightly acidic electrolyzed water (SAEW) against food pathogens on selected fresh ready-to-eat (RTE) vegetables and sprouts was evaluated and compared to sodium hypochlorite (NaOCl) solution. RTE vegetables and sprouts were dip-inoculated with Escherichia coli (E. coli) and Salmonella spp. and dip-treated with SAEW, NaOCl solution for 5 min. SAEW treatment significantly (p < 0.05) reduced the total aerobic mesophilic bacteria from Chinese celery, lettuce and daikon sprouts by 2.7, 2.5 and 2.45 log₁₀CFU/g, respectively relative to un-treated. Pathogens were significantly (p < 0.05) reduced from Chinese celery, lettuce and daikon sprouts by 2.7, 2.8 and 2.8 log₁₀CFU/g (E. coli) and 2.87, 2.91 and 2.91 log₁₀CFU/g (Salmonella spp.), respectively following a SAEW treatment. SAEW and NaOCl solution showed no significant sanitization difference (p > 0.05). Results demonstrate that SAEW at low chlorine concentration and a near neutral pH is a potential non-thermal food sanitizer that could represent an alternative to NaOCl solution and would reduce the amount of free chlorine used in fresh-cut vegetables industry, since the same microbial reduction as NaOCl solution is obtained.     

A novel common primer multiplex PCR (CP-M-PCR) method for the simultaneous detection of meat species

A novel common primer multiplex PCR (CP-M-PCR) was applied to detect four kinds of meats (chicken, cattle, pig and horse) as raw materials. A common adapter was designed in the 5′-end of species-specific reverse primers which matched with the species-specific DNA sequences for each species and also used as the common primer (CP). CP-M-PCR primers were designed to uncover different length fragments of 239, 292, 412, and 451 bp from chicken, cattle, pig and horse meats, respectively. The bands of specific DNA fragments amplified by CP-M-PCR method still appeared until the concentration of species-specific primers diluted to 0.015 pmol and primer sensitivity was increased by 100 times compared with conventional multiplex PCR without CP. CP-M-PCR detection limit of the DNA samples was 0.1 ng (36.4 copies) for single kind of meat as well as four kinds of meats. CP-M-PCR method simplified the PCR reaction system and conquered the disparate amplified efficiency from different primers. The CP-M-PCR method could be widely applied in practical detection for simultaneous identification of other meat species and their products.     

Prevalence of foodborne pathogens in Turkish Van otlu (Herb) cheese

The survey was conducted on 50 unripened Van otlu cheese samples obtained in Van and Hakkari markets at retail level to determine the microbial characteristics with special emphasis on Staphylococcus aureus, Escherichia coli, E. coli O157:H7 and Salmonella spp. The results revealed that S. aureus and E. coli were present in extremely high numbers, with a mean 6.10 and 3.68 log CFU/g, respectively. S. aureus was found in all samples ranging from 2.48 to 7.15 log CFU/g and was present in more than 5.0 × 10⁵ CFU/g in 54% of the samples whereas E. coli was found in 62% of the samples. None of the samples contained E. coli O157:H7; but 3 of the 50 samples had Salmonella spp. The results indicate that Van otlu cheese presents a potential hazard for public health; and the necessary precaution will have to be taken to improve the sanitary practices and cheese manufacturing technique.     

Improvement of the food safety of low acid fermented sausages by enterocins A and B and high pressure

Fermented sausage technology involves a sequence of hurdles that appear along the ripening process. A wide variety of fermented sausages are manufactured worldwide based on the concept of reduction of pH and/or water activity. In low acid fermented sausages the absence of high acidification can be balanced by the application of additional hurdles such as bacteriocins and/or high hydrostatic pressure (HHP). The addition of enterocins A and B to raw-sausages spiked with 3 log CFU/g of Salmonella, Listeria monocytogenes and Staphylococcus aureus showed an immediate reduction in the counts of L. monocytogenes due to the enterocins, while Salmonella was more affected by the endogenous hurdles associated with the ripening process. The application of an HHP treatment of 400 MPa at the end of ripening produced an immediate reduction in the counts of Salmonella but not in L. monocytogenes or S. aureus. During storage of the low acid sausages (fuets) at room temperature and at 7 °C, counts of Salmonella and L. monocytogenes progressively decreased in all batches although the decrease was faster in the pressurized ones stored at room temperature. At the end of storage, Salmonella was <1 log CFU/g in all the batches but only the combination of enterocins and HHP could reduce the counts of L. monocytogenes to this level. Neither the ripening process, the enterocins nor the pressurization could control the levels of S. aureus.