Survival and growth behavior of common foodborne pathogens in falafel paste under different storage temperatures.

Falafel is a popular breakfast food in the Middle East that has been recently involved in several outbreaks of foodborne illnesses. The aim of the study was to explore the growth behavior of Salmonella enterica, Escherichia coli O157:H7, Shigella sonnie, Shigella flexneri, Listeria monocytogenes and Staphylococcus aureus in falafel paste (FP) under different storage temperatures (4, 10, or 24 °C) for 14 days. FP (pH = 6.2, aw = 0.96) was inoculated with 5.0 to 6.0 log CFU/g of each of the pathogens separately. Salmonella spp. significantly declined by 1.5 log at 4 °C but grew significantly by ca. 2 and 4 log at 10 and 24 °C, respectively after 14 days. E. coli O157:H7 significantly increased (4.5 log) in FP when stored under 24 °C and survived at a level of ~105 CFU/g at 10 °C. Comparatively, Sh. sonnie and Sh. flexneri showed a better survival pattern in FP stored under 4 °C and grew (˃ 3 log) after 5 days at 10 and 24 °C. L. monocytogenes was capable of growing by 1.9 and 4.3 log after 14 d days and by 3.9 log after 3 days at 4, 10, or 24 °C, respectively. No significant decline in S. aureus counts at 4 and 10 °C occurred, however, it increased significantly to ˃ 7 log CFU/g at 24 °C. Total mesophilic count and yeast and mold count reached to spoilage levels (˃107 CFU/g) in un-inoculated FP after 1 and 3 days of storage at 24 and 10 °C, respectively. FP could support the growth of common foodborne pathogens and hence it is recommended to utilize natural antimicrobials in FP and keep the product under refrigeration (4 °C) to preclude the growth of vegetative foodborne pathogens.

Effect of gamma irradiation against microbial spoilage of hummus preserved under refrigerated storage.

The current study aimed to evaluate the preservation of hummus packed in laminated pouches to extend its shelf life for a period of 35 d using a combined treatment of gamma irradiation and refrigerated storage. The effects of different irradiation doses (0, 0.1, 1.5 and 2.5 kGy) and refrigeration (4 °C) against microbial spoilage were investigated. Microbial criteria of total mesophilic aerobic counts, lactic acid bacterial counts, and yeasts and molds decreased significantly (P ≤ 0.05) in a dose dependent manner. Furthermore, dose of 2.5 kGy decreased the lactic acid bacteria counts to undetectable levels for the first 10 d of storage. Although microbial load increased during 35 d of refrigerated storage, samples treated with doses of 1.5 and 2.5 kGy showed a good microbial quality that complies with microbial limits set by national and international authorities for ready to eat foods. Furthermore, Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, Enterococcus faecalis and coliform bacteria were not detected in the irradiated samples as well as in the controls. It was observed that a dose ≥1.5 kGy combined with refrigerated storage at 4 °C could be used to extend the shelf life of hummus samples in laminated pouches up to 35 d.

Effect of acidic electrolyzed water-induced bacterial inhibition and injury in live clam (Venerupis philippinarum) and mussel (Mytilus edulis).

The effect of acidic electrolyzed water (AEW) on inactivating Escherichia coli O104:H4, Listeria monocytogenes, Aeromonas hydrophila, Vibrio parahaemolyticus and Campylobacter jejuni in laboratory contaminated live clam (Venerupis philippinarum) and mussel (Mytilus edulis) was investigated. The initial levels of bacterial contamination were: in clam 4.9 to 5.7log10CFU/g, and in mussel 5.1 to 5.5log10CFU/g. Two types of AEW were used for treatment time intervals of 1 and 2h: strong (SAEW) with an available chlorine concentration (ACC) of 20mg/L, pH=3.1, and an oxidation-reduction potential (ORP) of 1150mV, and weak (WAEW) at ACC of 10mg/L, pH=3.55 and ORP of 950mV. SAEW and WAEW exhibited significant inhibitory activity against inoculated bacteria in both shellfish species with significant differences compared to saline solutions treatments (1-2% NaCl) and untreated controls (0h). SAEW showed the largest inhibitory activity, the extent of reduction (log10CFU/g) ranged from 1.4-1.7 for E. coli O104:H4; 1.0-1.6 for L. monocytogenes; 1.3-1.6 for A. hydrophila; 1.0-1.5 for V. parahaemolyticus; and 1.5-2.2 for C. jejuni in both types of shellfish. In comparison, significantly (P<0.05) lower inhibitory effect of WAEW was achieved compared to SAEW, where the extent of reduction (log10CFU/g) ranged from 0.7-1.1 for E. coli O104:H4; 0.6-0.9 for L. monocytogenes; 0.6-1.3 for A. hydrophila; 0.7-1.3 for V. parahaemolyticus; and 0.8-1.9 for C. jejuni in both types of shellfish. Among all bacterial strains examined in this study, AEW induced less bacterial injury (~0.1-1.0log10CFU/g) and more inactivation effect. This study revealed that AEW (10-20mg/L ACC) could be used to reduce bacterial contamination in live clam and mussel, which may help control possible unhygienic practices during production and processing of shellfish without apparent changes in the quality of the shellfish.

Efficacy of Neutral Electrolyzed Water, Quaternary Ammonium and Lactic Acid-Based Solutions in Controlling Microbial Contamination of Food Cutting Boards Using a Manual Spraying Technique.

Bactericidal activity of neutral electrolyzed water (NEW), quaternary ammonium (QUAT), and lactic acid-based solutions was investigated using a manual spraying technique against Salmonella Typhimurium, Escherichia coli O157:H7, Campylobacter jejuni, Listeria monocytogenes and Staphylococcus aureus that were inoculated onto the surface of scarred polypropylene and wooden food cutting boards. Antimicrobial activity was also examined when using cutting boards in preparation of raw chopped beef, chicken tenders or salmon fillets. Viable counts of survivors were determined as log10 CFU/100 cm(2) within 0 (untreated control), 1, 3, and 5 min of treatment at ambient temperature. Within the first minute of treatment, NEW and QUAT solutions caused more than 3 log10 bacterial reductions on polypropylene surfaces whereas less than 3 log10 reductions were achieved on wooden surfaces. After 5 min of treatment, more than 5 log10 reductions were achieved for all bacterial strains inoculated onto polypropylene surfaces. Using NEW and QUAT solutions within 5 min reduced Gram-negative bacteria by 4.58 to 4.85 log10 compared to more than 5 log10 reductions in Gram-positive bacteria inoculated onto wooden surfaces. Lactic acid treatment was significantly less effective (P < 0.05) compared to NEW and QUAT treatments. A decline in antimicrobial effectiveness was observed (0.5 to <2 log10 reductions were achieved within the first minute) when both cutting board types were used to prepare raw chopped beef, chicken tenders or salmon fillets.

Effect of oxygen stress on growth and survival of Clostridium perfringens, Campylobacter jejuni, and Listeria monocytogenes under different storage conditions.

This study investigated the growth and survival of three foodborne pathogens (Clostridium perfringens, Campylobacter jejuni, and Listeria monocytogenes) in beef (7% fat) and nutrient broth under different oxygen levels. Samples were tested under anoxic (<0.5%), microoxic (6 to 8%), and oxic (20%) conditions during storage at 7 °C for 14 days and at 22 °C for 5 days. Two initial inoculum concentrations were used (1 and 2 log CFU per g of beef or per ml of broth). The results show that C. perfringens could grow in beef at 22 °C, with an increase of approximately 5 log under anoxic conditions and a 1-log increase under microoxic conditions. However, C. perfringens could not survive in beef held at 7 °C under microoxic and oxic storage conditions after 14 days. In an anoxic environment, C. perfringens survived in beef samples held at 7 °C, with a 1-log reduction. A cell decline was observed at 2 log under these conditions, with no surviving cells at the 1-log level. However, the results show that C. jejuni under microoxic conditions survived with declining cell numbers. Significant increases in L. monocytogenes (5 to 7 log) were observed in beef held at 22 °C for 5 days, with the lowest levels recovered under anoxic conditions. L. monocytogenes in refrigerated storage increased by a factor of 2 to 4 log. It showed the greatest growth under oxic conditions, with significant growth under anoxic conditions. These findings can be used to enhance food safety in vacuum-packed and modified atmosphere-packaged food products.

Antifungal effectiveness of potassium sorbate incorporated in edible coatings against spoilage molds of apples, cucumbers, and tomatoes during refrigerated storage.

Predominant spoilage molds of fresh apples, cucumbers, and tomatoes stored at 4 °C were isolated and examined for resistance to potassium sorbate (PS) incorporated in polysaccharide edible coatings. The isolated molds were Penicillium expansum, Cladosporium herbarum, and Aspergillus niger from apples. P. oxalicum and C. cucumerinum were isolated from cucumbers and P. expansium and C. fulvum from tomatoes. Guar gum edible coating incorporated with PS was the most effective mold inhibitor, significantly (P<0.05) reducing the isolated spoilage molds for 20, 15, and 20 d of storage at 4 °C on apples, cucumbers, and tomatoes, respectively. PS incorporated into pea starch edible coating was less effective and selectively inhibited the isolated mold species, causing significant (P<0.05) reduction in mold on apples, cucumbers, and tomatoes counts for 20, 10 to 15, and 15 to 20 d of storage at 4 °C, respectively. The isolated mold species exhibited different resistances to PS incorporated in the edible coatings. The greatest inhibition (2.9 log CFU/g) was obtained with C. herbarum on apples and the smallest (1.1 log CFU/g) was with P. oxalicum on cucumbers and the other isolated mold species exhibited intermediate resistance. The coatings tested, in general, inhibited molds more effectively on apples than on tomatoes and cucumbers. Addition of PS to pea starch and guar gum, edible coatings improved the antifungal activity of PS against isolated spoilage molds on apples, cucumbers, and tomatoes. PS inhibition was most effective against C. herbarum on apples and least effective against P. oxalicum on cucumbers.

Effect of refrigerated and frozen storage on the survival of Campylobacter jejuni in cooked chicken meat breast.

This experimental work aimed to examine the survivability of Campylobacter jejuni in cooked chicken breast under several conditions: storage for 1, 3, and 7 d at refrigerated temperatures (4 °C) and for 20 d at frozen temperatures (-18 °C). In addition, storage at ambient temperature (26 to 28 °C) was involved. Chicken samples were inoculated with a mixed culture of C. jejuni strains (ATCC: 29428 and 33219) of known concentrations (50 and 500 CFU/g). Bacterial cells were recovered and enumerated using standard procedure (Preston method). Bacteria were not detected in the majority of samples stored at ambient temperature. Refrigeration reduced survivals in 95, 90, and 77.5% for samples inoculated with 500 CFU/g and kept for 1, 3, and 7 d, respectively. The maximum reduction reached 1 log(10) cycle for all refrigeration durations. It was observed that bacteria died in 17.5% of samples kept for 7 d at 4 °C. However, survivors in samples inoculated with 50 CFU/g were not detected in 50, 65, and 55% of samples kept for 1, 3, and 7 d, respectively. Freezing rendered survivors not detectable in 70% of samples inoculated with 50 CFU/g, while survived viable counts were reduced in 92.5% of samples inoculated with 500 CFU/g. These findings suggested that C. jejuni could be killed or just sublethally injured with or without reduction in viable counts under the investigated storage temperatures, which may indicate the ability of this bacterium to survive in chicken meat stored under refrigerated and frozen conditions.

Using of infrared spectroscopy to study the survival and injury of Escherichia coli O157:H7, Campylobacter jejuni and Pseudomonas aeruginosa under cold stress in low nutrient media.

The inactivation and sublethal injury of Escherichia coli O157:H7, Campylobacter jejuni and Pseudomonas aeruginosa at three temperatures (22 °C, 4 °C and -18 °C) were studied using traditional microbiological tests and mid-infrared spectroscopy (4000-400 cm(-1)). Bacteria were cultivated in diluted nutrient matrices with a high initial inoculation (∼10(7) CFU/ml) levels. Both E. coli O157:H7 and P. aeruginosa survived and cell numbers increased at 22 °C for 5 days while C. jejuni numbers decreased one log(10) CFU/ml. A two log CFU/ml decrease was observed for the three pathogens held at 4 °C for 12 days. C. jejuni survived poorly following incubation at -18 °C for 20 days while levels of E. coli O157:H7 and P. aeruginosa remained high (10(4) CFU/ml). Temperature stress response of microbes was observed by infrared spectroscopy in polysaccharide, protein, lipid, and nucleic acid regions and was strain specific. Level of cold injury could be predicted using cluster, discriminant function and class analog analysis models. Pathogens may produce oligosaccharides and potentially other components in response to stress as indicated by changes in spectral features at 1200-900 cm(-1) following freezing.

Survival of Escherichia coli O157:H7 and Campylobacter jejuni in bottled purified drinking water under different storage conditions.

Survival of Escherichia coli O157:H7 and Campylobacter jejuni that were separately inoculated into bottled purified drinking water was investigated during storage at 22, 4, and -18 °C for 5, 7, and 2 days, respectively. Two inoculation levels were used, 1 and 10 CFU/ml (10(2) and 10(3) CFU/100 ml). In samples inoculated with 10(2) CFU/100 ml, C. jejuni was not detectable (>2-log reduction) after storage under the conditions specified above. E. coli O157:H7 was detected on nonselective and selective media at log reductions of 1.08 to 1.25 after storage at 22 °C, 1.19 to 1.56 after storage at 4 °C, and 1.54 to 1.98 after storage at -18 °C. When the higher inoculation level of 10(3) CFU/100 ml was used, C. jejuni was able to survive at 22 and 4 °C, with 2.25- and 2.17-log reductions, respectively, observed on nonselective media. At these higher inoculation levels, E. coli O157:H7 was detectable at 22, 4, and -18 °C, with log reductions of 0.76, 0.97, and 1.21, respectively, achieved on nonselective media. Additionally, E. coli O157:H7 showed significant differences in culturability (P<0.05) on the nonselective and selective culture media under the different storage conditions, with storage at -18 °C for 2 days being the treatment most inhibiting. The percentage of sublethal injury of E. coli O157:H7 ranged from ∼33 to 75%, indicating that microbial examination of bottled water must be done carefully, otherwise false-negative results or underestimation of bacterial numbers could pose a health risk when low levels of pathogens are present.

Determination of total phenolic content and antioxidant capacity of onion (Allium cepa) and shallot (Allium oschaninii) using infrared spectroscopy.

Total phenolic content (TPC) and total antioxidant capacity (TAC) of four onion varieties (red, white, yellow and sweet) and shallot from selected locations (Washington, Idaho, Oregon, Texas and Georgia) were determined using Fourier transform infrared (FT-IR) spectroscopy (4000-400cm-1). The Folin-Ciocalteu (F-C) assay was used to quantify TPC and three assays were used to determine TAC, including 2,2-diphenyl-picrylhydrazyl (DPPH) assay, Trolox equivalent antioxidant capacity (TEAC) assay and ferric reducing antioxidant power (FRAP) assay. Partial least squares regression (PLSR) with cross-validation (leave-one-out) was conducted on onion and shallot extracts (n=200) and their corresponding F-C, DPPH, TEAC and FRAP values were employed to obtain four independent calibration models for predicting TPC and TAC for the extracts. Spectra from an extra 19 independent extracts were used as an external validation set for prediction. A correlation of r>0.95 was obtained between FT-IR predicted and reference values (by F-C, DPPH, TEAC and FRAP assay) with standard errors of calibration (SEC) and standard errors of cross-validation (SECV) less than 2.85, 0.35 and 0.45µmolTrolox/g FW of extracts for TEAC, FRAP and DPPH assay, respectively; and 0.36mggallic acid/g FW of extracts for the F-C assay. In addition, cluster analysis (principal component analysis (PCA)) and discriminant function analysis (DFA) could differentiate varieties of onions and shallot based upon infrared spectral features. Loading plots for the various chemometrics models indicated that hydroxyl and phenolic functional groups were most closely correlated with antioxidant capacity. The use of mid-infrared spectroscopy to predict the total antioxidant capacity of vegetables provides a rapid and precise alternative to traditional wet chemistry analysis.

Using Fourier transform infrared (FT-IR) absorbance spectroscopy and multivariate analysis to study the effect of chlorine-induced bacterial injury in water.

The effect of chlorine-induced bacterial injury on spectral features using Fourier transform infrared (FT-IR) absorbance spectroscopy was studied using a mixed bacterial culture of (1:1) ca. 500 CFU/mL each Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 15442 in 0.9% saline. Bacterial cells were treated with 0, 0.3, or 1.0 ppm of initial free chlorine (21 degrees C, 1 h of contact time). Chlorine-injured and dead bacterial cells retained the ATR spectral properties of uninjured or live cells in the region of C-O-C stretching vibrations of polysaccharides, indicative of the cell wall peptidoglycan layer and lipopolysaccharide outer leaflet. This confirms the observations of others that extensive bacterial membrane damage is not a key factor in the inactivation of bacteria by chlorine. The bactericidal effect of chlorine caused changes in the spectral features of bacterial ester functional groups of lipids, structural proteins, and nucleic acids, with apparent denaturation reflected between 1800 and 1300 cm (-1) for injured bacterial cells. Three-dimensional principal component analysis (PCA) showed distinct segregation and clustering of chlorine-treated and untreated cells. Cells exposed to chlorine at 0.3 or 1.0 ppm could be distinguished from the untreated control 73 and 80% of the time, respectively, using soft independent modeling of class analogy (SIMCA) analysis. This study suggests that FT-IR spectroscopy may be applicable for detecting the presence of injured and viable but not culturable (VBNC) waterborne pathogens that are underestimated or not discernible using conventional microbial techniques.

A comparative study between overlay method and selective-differential media for recovery of stressed Enterobacter sakazakii cells from infant formula.

This study compares the performance of different selective-differential media with the overlay method for recovery of stressed cells of Enterobacter sakazakii from infant formula milk (IFM). Five different selective-differential media were used in this study: OK medium, violet red bile agar (VRBA), Druggan-Forsythe-Iversen agar (DFI), Enterobacteriaceae enrichment (EE) agar, and fecal coliform agar (FCA). Tryptic soy agar supplemented with 0.1% sodium pyruvate (TSAP) was used as a control. The overlay method involved applying a thin layer (8ml) of each of the selective media onto TSAP after spreading a sample onto TSAP. Reconstituted IFM was inoculated by ca 1x10(7)CFU/ml of a mixture of four strains of E. sakazakii and subjected to different stress conditions: heat (55 degrees C for 10min), a freeze-thaw cycle (-20 degrees C for 24h, thawed at room temperature, frozen again at -20 degrees C, and thawed), acidic pH (pH 3.56 for 15min), alkaline pH (pH 11.04 for 15min), and desiccation (E. sakazakii was inoculated onto powdered IFM at a level of ca 1x10(6)CFU/g, held at 21 degrees C, water activity of the inoculated product was 0.29 and examined at 0, 15, and 30d). No major differences were noticed between the control (TSAP) and the overlay methods. However, the overlay method recovered significantly higher numbers of stressed E. sakazakii cells compared to selective-differential media. Also, the selective-differential media exhibited some variability in terms of their capabilities to recover stressed cells of E. sakazakii. Among all the examined selective-differential media, DFI performed better for recovering stressed E. sakazakii cells. This study suggests that the overlay method may serve as a potential alternative to direct selective plating for best recovery of E. sakazakii from IFM.

Rapid detection and identification of Pseudomonas aeruginosa and Escherichia coli as pure and mixed cultures in bottled drinking water using fourier transform infrared spectroscopy and multivariate analysis.

Fourier transform infrared (FT-IR) spectroscopy and multivariate analysis were used to identify Pseudomonas aeruginosa and Escherichia coli ATCC 25922 inoculated into bottled drinking water. Three inoculation treatments were examined: (i) E. coli ATCC 25922 (N = 3), (ii) P. aeruginosa (N = 3), and (iii) a 1:1 (v:v) mixed culture of both P. aeruginosa and E. coli ATCC 25922 (N = 3). The control treatment was noninoculated drinking water (N = 3). Second derivative transformation and loadings plots over the range of 1800-900 cm(-1) indicate variations in the following bacterial constituents: amide I band ca. 1650 cm(-1), amide II band ca. 1540 cm(-1), phosphodiester backbone of nucleic acids ca. 1242 and 1080 cm(-1), and polysaccharide compounds ca. 1050-950 cm(-1). Cells with the different treatments were clearly segregated from a mean centered principal component analysis. By using soft independent modeling of class analogy analysis, spectra from a given treatment could be correctly classified 83-88% of the time. These results suggest that FT-IR spectroscopy can determine whether a pure culture is present, in addition to confirming that this method can discriminate between closely related bacteria based on differences in biochemical and phenotypic characteristics that can be detected in this spectral region.

Fourier transform infrared spectroscopy, detection and identification of Escherichia coli O157:H7 and Alicyclobacillus strains in apple juice.

Fourier Transform Infrared (FT-IR) spectroscopy (4000-400 cm(-1)) combined with multivariate statistical methods were used to identify and detect Escherichia coli O157:H7 from Alicyclobacillus spp. recovered from apple juice. Four treatments and a control in triplicate experiments (N=3) were studied; the first three treatments of pasteurized apple juice were inoculated with E. coli O157:H7 ATCC 35150, Alicyclobacillus acidoterrestris 1016 and Alicyclobacillus spp. C-Fugi-6 respectively. The fourth treatment was a 1:1 (v:v) mixed culture of both A. acidoterrestris 1016 and Alicyclobacillus spp. C-Fugi-6. The control was uninoculated pasteurized apple juice. The second derivative transformation and loadings plot over the range of 1800-900 cm(-1) highlighted the most distinctive variations among bacterial spectra. Loadings 1 and 2 were distinctively representative of the bacterial spectral data and accounted for 73% of the total variability. Treatments were noticeably segregated with distinct clustering by principal component analysis (PCA). Using soft independent modeling of class analogy (SIMCA) analysis, 88.3% of (E. coli O157:H7 ATCC 35150) spectra, 75.0% of (A. acidoterrestris 1016) spectra, 88.3% of (Alicyclobacillus spp. C-Fuji-6) spectra, and 80.0% of the mixed culture of both Alicyclobacillus strains spectra were correctly classified. Using the spectral features of bacterial cellular constituents such as nucleic acids, proteins, phospholipids, peptidoglycan, and lipopolysaccharides from examined bacterial cells, pure and mixed cultures of Alicyclobacillus spp. cells, and the pathogenic E. coli cells could be detected in apple juice.

Discrimination of intact and injured Listeria monocytogenes by Fourier transform infrared spectroscopy and principal component analysis.

Fourier transform infrared spectroscopy (FT-IR, 4000-600 cm(-)(1)) was used to discriminate between intact and sonication-injured Listeria monocytogenes ATCC 19114 and to distinguish this strain from other selected Listeria strains (L. innocua ATCC 51742, L. innocua ATCC 33090, and L. monocytogenes ATCC 7644). FT-IR vibrational overtone and combination bands from mid-IR active components of intact and injured bacterial cells produced distinctive "fingerprints" at wavenumbers between 1500 and 800 cm(-)(1). Spectral data were analyzed by principal component analysis. Clear segregations of different intact and injured strains of Listeria were observed, suggesting that FT-IR can detect biochemical differences between intact and injured bacterial cells. This technique may provide a tool for the rapid assessment of cell viability and thereby the control of foodborne pathogens.