Effect of nisin in combination with EDTA, sodium lactate, and potassium sorbate for reducing Salmonella on whole and fresh-cut cantaloupet

Nisin (50 microg/ml), EDTA (0.02 M, disodium salt), sodium lactate (NaL, 2%), and potassium sorbate (KS, 0.02%) were tested individually and in various combinations as sanitizer treatments for reducing Salmonella on whole and fresh-cut cantaloupe. Whole cantaloupe and fresh-cut pieces were inoculated with a five-strain cocktail of Salmonella to give 4.76 +/- 0.23 log CFU/cm2 and 3.42 +/- 0.13 log CFU/g, respectively. Inoculated whole melons and fresh-cut pieces were stored at 5 degrees C for 7 days. Washing treatments were applied to inoculated whole melons at days 0, 3, and 7 of storage, and surviving bacterial populations were determined. The effect of the washing treatments on transfer of Salmonella to fresh-cut pieces prepared immediately after treatment was also determined. Directly inoculated fresh-cut pieces were treated at day 0, and surviving bacteria were enumerated at days 0, 3, and 7 of storage. The combination treatments of nisin-EDTA, nisin-NaL, nisin-KS, NaL-KS, and nisin-NaL-KS all resulted in reductions of approximately 3 log CFU/cm2 at day 0 for whole melons. When tested alone, all compounds, along with water washes, were ineffective. After 3 and 7 days of storage, the five combination washing treatments were less effective, resulting in reductions of approximately 2 log CFU/cm2. None of the combination treatments completely eliminated transfer of pathogen survivors to fresh-cut pieces. The combination treatments nisin-NaL, nisin-KS, NaL-KS, and nisin-NaL-KS, but not nisin-EDTA, gave significant (P < 0.05) reductions of Salmonella directly inoculated onto fresh-cut pieces. Washing with nisin-NaL-KS was significantly (P < 0.05) more effective than the other three combination treatments, resulting in a reduction of 1.4 CFU/g. Inhibition by the four effective treatments carried over from day 0 through day 7 of storage, with no increase in the population of Salmonella on the stored fresh-cut pieces. Sensory evaluations indicated that treatment of fresh-cut pieces with nisin-NaL and NaL-KS, but not nisin-KS or nisin-NaL-KS, were acceptable in terms of appearance, odor, and overall acceptability. After the required regulatory approval, treatment of whole cantaloupe with nisin in combination with EDTA, NaL, KS, or NaL and KS and of fresh-cut pieces with nisin-NaL or NaL-KS could help ensure the microbiological safety of fresh-cut cantaloupe.     

Combined efficacy of nisin and pediocin with sodium lactate, citric acid, phytic acid, and potassium sorbate and EDTA in reducing the Listeria monocytogenes population of inoculated fresh-cut produce

The inability of chlorine to completely inactivate human bacterial pathogens on whole and fresh-cut produce suggests a need for other antimicrobial washing treatments. Nisin (50 microg/ml) and pediocin (100 AU/ml) individually or in combination with sodium lactate (2%), potassium sorbate (0.02%), phytic acid (0.02%), and citric acid (10 mM) were tested as possible sanitizer treatments for reducing the population of Listeria monocytogenes on cabbage, broccoli, and mung bean sprouts. Cabbage, broccoli, and mung bean sprouts were inoculated with a five-strain cocktail of L. monocytogenes at 4.61, 4.34, and 4.67 log CFU/g, respectively. Inoculated produce was left at room temperature (25 degrees C) for up to 4 h before antimicrobial treatment. Washing treatments were applied to inoculated produce for 1 min, and surviving bacterial populations were determined. When tested alone, all compounds resulted in 2.20- to 4.35-log reductions of L. monocytogenes on mung bean, cabbage, and broccoli, respectively. The combination treatments nisin-phytic acid and nisin-pediocin-phytic acid caused significant (P < 0.05) reductions of L. monocytogenes on cabbage and broccoli but not on mung bean sprouts. Pediocin treatment alone or in combination with any of the organic acid tested was more effective in reducing L. monocytogenes populations than the nisin treatment alone. Although none of the combination treatments completely eliminated the pathogen on the produce, the results suggest that some of the treatments evaluated in this study can be used to improve the microbial safety of fresh-cut cabbage, broccoli, and mung bean sprouts.     

Comparison of different washing treatments for reducing pathogens on orange surfaces and for preventing the transfer of bacterial pathogens to fresh-squeezed orange juice

The objectives of this study were to compare the effectiveness of various washing treatments for reducing Escherichia coli O157:H7, Salmonella sp., and Listeria monocytogenes populations on orange surfaces and to measure the effect of some of these treatments in preventing the transfer of pathogens during juice extraction. Orange surfaces inoculated with L. monocytogenes or a mixture of E. coli O157:H7 and Salmonella Typhimurium were washed by water spray and then sprayed with or dipped in water at 80°C for 1 min, 70% ethanol for 15, 30, or 45 s or 1, 2, or 4 min, 2 or 4% lactic acid solution at 55°C for 15, 30, or 45 s or 1, 2, or 4 min, or 200 mg/liter hypochlorite at pH 6.5 or 10 for 15 s. The surviving populations of these pathogens on the oranges were enumerated after each treatment. In a further stage, the ability of these pathogens to be transferred to the juice during extraction was tested. Juice was obtained from inoculated oranges that were subjected to selected treatments using chlorine, lactic acid, ethanol, and hot water as described above, and then bacterial counts in orange juice were determined. The application of these treatments reduced the populations of pathogens on orange surfaces by 1.9 to >4.9 log, 1.9 to >4.6 log, and 1.4 to 3.1 log cycles for E. coli O157:H7, Salmonella Typhimurium, and L. monocytogenes, respectively. The treatments using hot water or lactic acid showed greater reductions than other treatments. The time, antimicrobial concentration, and form of application affected the bacterial reduction. All treatments resulted in undetectable counts in the juice. Nevertheless, pathogens were recovered by the enrichment-plating method. Treatment of oranges before juice extraction may reduce the risk associated with consuming orange juice.     

Behavior of Listeria monocytogenes inoculated on cantaloupe surfaces and efficacy of washing treatments to reduce transfer from rind to fresh-cut pieces

Attachment and survival of Listeria monocytogenes on external surfaces (rind) of inoculated cantaloupe, resistance of the surviving bacteria to chlorine or hydrogen peroxide treatments, transfer of the pathogen from unsanitized and sanitized rinds to fresh-cut tissues during cutting and growth, and survival of L. monocytogenes on fresh-cut pieces of cantaloupe were investigated. Surface treatment with 70% ethanol to reduce the native microflora on treated melon, followed by immersion in a four-strain cocktail of L monocytogenes (10(8) CFU/ml) for 10 min, deposited 4.2 log10 CFU/cm2 and 3.5 log10 CFU/cm2 of L monocytogenes on treated and untreated cantaloupe rinds, respectively. L. monocytogenes survived on the treated or untreated cantaloupe rinds for up to 15 days during storage at 4 and 20 degrees C, but populations declined by approximately 1 to 2 log10 CFU/cm2. Fresh-cut pieces prepared from inoculated whole cantaloupes stored at 4 degrees C for 24 h after inoculation were positive for L. monocytogenes. Washing inoculated whole cantaloupes in solutions containing 1,000 ppm of chlorine or 5% hydrogen peroxide for 2 min at 1 to 15 days of storage at 4 degrees C after inoculation resulted in a 2.0- to 3.5-log reduction in L. monocytogenes on the melon surface. Fresh-cut pieces prepared from the sanitized melons were negative for L. monocytogenes. After direct inoculation onto fresh-cut pieces, L. monocytogenes survived, but did not grow, during 15 days of storage at 4 degrees C. Growth was evident by 4 h of storage at 8 and 20 degrees C. It is concluded that sanitizing with chlorine or hydrogen peroxide has the potential to reduce or eliminate the transfer of L. monocytogenes on melon surfaces to fresh-cut pieces during cutting.     

Use of hydrogen peroxide detection strips to determine the extent of pasteurization in whole milk

Commercially available hydrogen peroxide (H₂O₂) detection strips were shown to be effective in determining known levels of H₂O₂ in milks where the lactoperoxidase system (LPS) had been inactivated. In addition, the strips were evaluated for determination of residual H₂O₂ in LPS-activated milks, following a range of heat treatments. The detection of residual H₂O₂ corresponded with results of zero lactoperoxidase activity. Hence, the detection strips offer a simple and accurate method for detecting the absence of lactoperoxidase, and therefore can be used to determine whether the milk has been subjected to overpasteurization. The technique provides a more convenient method than the standard procedure based on oxidation of 1,4-phenylenediamine.     

Fate of Listeria monocytogenes inoculated onto the surface of model Turkey frankfurter pieces treated with zein coatings containing nisin, sodium diacetate, and sodium lactate at 4 degrees C

The antimicrobial effects of zein coatings containing nisin, sodium lactate, and sodium diacetate against Listeria monocytogenes on turkey frankfurters at 4 degrees C were determined. Our objectives were to determine whether zein, nisin, lactate, and diacetate alone or in combination could control the growth of L. monocytogenes on full-fat turkey frankfurters at 4 degrees C and to determine whether lactate or diacetate had any synergistic effect on the activity of nisin. Turkey frankfurter pieces surface inoculated with L. monocytogenes strain V7 were treated with zein-ethanol-glycerol (ZEG), zein-propylene-glycol (ZPR), ethanol-glycerol (EG), propylene glycol (PR), nisin (N), sodium lactate (L), or sodium diacetate (D) alone or in combination. Over 28 days, treatment with N or D alone reduced L. monocytogenes counts on frankfurters by 6.6 or 6.3 log CFU/g, respectively. N-D treatment reduced L. monocytogenes by 6 log CFU/g. The zein solvents EG and PR reduced L. monocytogenes by about 5.6 and 5.2 log CFU/g, respectively, similar to the results obtained with ZEG and ZPR, which suggests that zein powder per se had no antimicrobial activity. After 28 days, ZEG-N-D, ZEG-N-D-L, ZPR-N-D, and ZPR-N-D-L yielded no detectable CFU. L alone was ineffective. No synergies were observed. N and D when used singly and the combinations of N-D, ZEG-N-D, ZEG-N-D-L, ZPR-N-D, ZPR-N-D-L, EG, and PR were effective as inhibitors of the growth of recontaminating L. monocytogenes cells on full-fat turkey frankfurters.     

Effectiveness of potassium lactate and sodium diacetate in combination with irradiation to control Listeria monocytogenes on frankfurters

ABSTRACT:  The use of antimicrobial ingredients in combination with irradiation is an effective antilisterial intervention strategy for ready-to-eat meat products. Microbial safety was evaluated for frankfurters formulated with 0% or 3% added potassium lactate/sodium diacetate solution and inoculated with Listeria monocytogenes before or after treatment with irradiation (0, 1.8, or 2.6 kGy). Frankfurters were stored aerobically or vacuum packaged and L. mo nocytogenes counts and APCs were determined while refrigerated. The incorporation of lactate/diacetate with or without irradiation had a strong listeriostatic effect for aerobically stored frankfurters. Outgrowth was suppressed and counts were not different from initial counts (5.2 log CFU/frank compared with 5.0 log CFU/frank); however, those without the additive increased steadily (5.4 to 9.3 log CFU/frank). Irradiation treatments alone had higher L. monocytogenes counts after 3 wk. For vacuum-packaged frankfurters, both the addition of lactate/diacetate and irradiation were effective at controlling growth after 8 wk. Large and incremental reductions in total counts were seen for irradiation treatments. Initial counts were reduced by 3 log CFU with the application of 1.8 kGy while 2.6 kGy decreased counts over 5 log CFU. These reductions were maintained throughout storage for lactate/diacetate-treated frankfurters. By 8 wk, L. monocytogenes counts on 1.8 and 2.6 kGy irradiated frankfurters without lactate/diacetate increased to 7.43 and 6.13 log CFU, respectively. Overall, lactate/diacetate retarded the outgrowth of L. monocytogenes on frankfurters throughout aerobic storage and the combination of irradiation and 3% lactate/diacetate reduced and retarded growth of L. monocytogenes , especially during the last 2 wk of vacuum-packaged storage.     

Reductions of Salmonella enterica on chicken breast by thymol, acetic acid, sodium dodecyl sulfate or hydrogen peroxide combinations as compared to chlorine wash

Poultry products are important vehicles for Salmonella transmission to humans and have been incriminated in several Salmonella outbreaks. Thymol (THY) from thyme oil has wide inhibitory effects against foodborne pathogens including Salmonella, and has shown great potential as a natural alternative to chlorine. In order to improve the cost-effectiveness of thymol-based washing solutions, formulas of THY with combination of organic acid or surfactant were developed and their efficacies to reduce Salmonella on chicken breast were investigated in the current study. Surface-inoculated chicken breasts were washed with the two thymol-based washing solutions: 0.2 mg/mL THY+5% (w/v) sodium dodecyl sulfate (SDS)+2 mg/mL acetic acid (AA) or 0.2 mg/mL THY+2 mg/mL AA for 2 min. Both solutions achieved around 2.2 log reductions of Salmonella on chicken breast and their efficacy was comparable to log reduction obtained by 200 ppm chlorine washing. Addition of SDS did not result in more log reduction of Salmonella on chicken meat samples. More than 3.3 log reduction in the used THY washing solutions was determined and it was similar to log reduction from the spent chlorine solution. None of these antimicrobial agents changed the pH and texture values of chicken breasts. Therefore, 0.2 mg/mL THY+2 mg/mL AA has great potential to be a natural alternative to chlorine-based washing solution for reducing Salmonella contamination on chicken breast meat.     

Decontamination with lactic acid/sodium lactate buffer in combination with modified atmosphere packaging effects on the shelf life of fresh poultry.

The effect of the treatment with various concentrations (2%, 5%, 7.5% and 10% w/v) of lactic acid/sodium lactate buffer (pH 3.0) combined with modified atmosphere packaging (MAP) (90% CO2/10% O2) on the shelf life and organoleptic quality of fresh chicken legs stored at 6 degrees C was investigated. The CO2 concentration of all samples packed in modified atmosphere (MA) decreased during the first 3 days of storage, followed by a gradual increase after the third day, while O2 showed a corresponding decrease. The buffering capacity of the buffer systems seem to be sufficient to maintain a low pH of the skin during storage. Legs treated with 2, 5, 7.5, and 10% (w/v) lactic acid/sodium lactate buffer (pH 3.0) combined with MAP have a shelf life at 6 degrees C of 14, 15, 16 and 17 days, respectively. The shelf life when the product was not treated with lactic acid was 1, 2.3 and 4 days shorter, respectively.     

Effects of sodium bicarbonate containing traces of citric acid in combination with sodium chloride on yield and some properties of white shrimp (Penaeus vannamei) frozen by shelf freezing, air-blast and cryogenic freezing

Effects of sodium bicarbonate with traces of citric acid in combination with sodium chloride on yield, freezing time, freezing rate, freezing loss and cutting force of white shrimp frozen by shelf, air-blast and cryogenic freezing with/without precooking were investigated. Shelf freezing was done at −40 °C ± 2 °C while air-blast freezing was carried out at −35 °C ± 2 °C, and cryogenic freezing was done at −35 °C, −40 °C and −60 °C. The freezing loss in the non-treated samples was 8.25, 4.6-5.84 and 1.92-3.48 g/100 g fresh shrimp for peeled samples frozen without precooking and increased to 21.85, 17.54-26.97, 17.92-20.31 g/100 g fresh shrimp in the precooked samples frozen by shelf, air-blast and cryogenic freezing, respectively. The treatment of sodium bicarbonate containing traces of citric acid at 4 g/100 ml with sodium chloride at 3 g/100 ml lead to the increase of yield thus reduced the freezing loss by about 6.83-10.28 and 6.41-12.4 g/100 g fresh shrimp for the frozen-thawed samples frozen as uncooked and cooked products, respectively. The toughening of shrimp was observed while sodium bicarbonate containing traces of citric acid treatment with sodium chloride could reduce the texture change occurred during the freezing.     

Antimicrobial susceptibility of isolated pathogens from patients with contact lens-related bacterial keratitis in Crete,Greece: A ten-year analysis

PurposeTo evaluate the bacterial spectrum and antimicrobial susceptibilities of pathogens isolated from contact lens-related bacterial keratitis cases in a large academic Greek hospital.MethodsAll adult patients with positive corneal scrapings or contact lens culture between 2007 and 2016 at the University Hospital of Heraklion, Greece, were retrospectively identified through a local microbiology database and their medical records were reviewed.ResultsA total of 240 isolates were recovered from 131 patients with culture-proven contact lens-associated bacterial keratitis. The most common microorganism identified was Serratia marcescens (17.1% of total isolates), followed by Pseudomonas aeruginosa, Klebsiella spp. and coagulase-negative staphylococci (CoNS). Rates of aztreonam-resistant P. aeruginosa and erythromycin-resistant CoNS decreased in recent years, while the decrease in oxacillin-resistant CoNS was statistically significant (p=0.009). More than 90% of the isolated organisms (S. marcescens, P. aeruginosa, Klebsiella spp. and CoNS) were susceptible to ciprofloxacin or gentamicin.ConclusionGram-negative microorganisms are the most common causative pathogens of contact lens-related keratitis in the region of Crete. Topical antibacterials containing quinolones or gentamicin represent an effective empirical treatment for the majority of the cases. This is quite encouraging, considering that the present study was conducted in a country characterised by high antimicrobial resistance rates. However, culture-driven antimicrobial treatment is mandatory for this sight-threatening infection.     

Efficiency of sodium hypochlorite, fumaric acid, and mild heat in killing native microflora and Escherichia coli O157:H7, Salmonella typhimurium DT104, and Staphylococcus aureus attached to fresh-cut lettuce

The effect of the disinfectant sodium hypochlorite (NaClO), with or without mild heat (50 degrees C) and fumaric acid, on native bacteria and the foodborne pathogens Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella Typhimurium DT104 attached to iceberg lettuce leaves was examined. The retail lettuce examined consistently harbored 6 to 7 log CFU/g of native bacteria throughout the study period. Inner leaves supported 1 to 2 log CFU/g fewer bacteria than outer leaves. About 70% of the native bacterial flora was removed by washing five times with 0.85% NaCl. S. aureus, E. coli, and Salmonella allowed to attach to lettuce leaves for 5 min were more easily removed by washing than when allowed to attach for 1 h or 2 days, with more S. aureus being removed than E. coli or Salmonella Typhimurium. An increase of time for attachment of pathogens from 5 min to 2 days leads to decreased efficiency of the washing and sanitizing treatment. Treatment with fumaric acid (50 mM for 10 min at room temperature) was the most effective, although it caused browning of the lettuce, with up to a 2-log reduction observed. The combination of 200 ppm of sodium hypochlorite and mild heat treatment at 50 degrees C for 1 min reduced the pathogen populations by 94 to 98% (1.2- to 1.7-log reduction) without increasing browning.     

Use of an extremely polar 100-m column in combination with a cyanoalkyl polysiloxane column to complement the study of milk fats with different fatty acid profiles

A detailed dairy fat fatty acid (FA) profile can generally be achieved by gas chromatography using capillary columns with polar cyanoalkyl polysiloxane stationary phases (CPSP). However some compounds overlap and cannot be discriminated after a single run. In this work a new, extremely polar phase column (SLB-IL111) was used to complement the analysis carried out by a CPSP column. Separation of the FAs in a variety of milk fats using both a 100-m SLB-IL111 column and a 100-m CP-Sil 88 column was carried out and a more complete profile was established. The combination of results was useful to discriminate FA pairs, i.e., 10:1/11:0, cis-10 12:1/13:0, iso 17:0/trans-9 16:1, anteiso 17:0/cis-9 16:1, trans-16 18:1/cis-14 18:1, trans-11 cis-15 18:2/trans-10 cis-15 18:2 and trans-7 cis-9 18:2/cis-9 trans-11 18:2. Furthermore trans-8 and trans-13/14 18:1 isomers were also well discriminated. The use of both columns allowed detection in milk of some lactone and keto FAs.     

Use of logistic regression with dummy variables for modeling the growth-no growth limits of Saccharomyces cerevisiae IGAL01 as a function of sodium chloride, acid type, and potassium sorbate concentration according to growth media

A global logistic model was used to study the effects of both quantitative variables (NaCl, acid, and potassium sorbate concentrations) and dummy variables (laboratory medium or brine, and citric, lactic, or acetic acids) on growth of Saccharomyces cerevisiae IGAL01. The deduced equations, with the significant coefficients selected by a backward stepwise procedure, allowed estimations of the simultaneous comparison of behaviors of levels of the qualitative variables as a function of the quantitative variables and the development of the growth-no growth limits according to laboratory medium or brine and the different types of acidifying agents. The S. cerevisiae growth region in yeast malt glucose peptone broth was always wider than that in brine, in which this yeast was inhibited by 0.03% potassium sorbate and 6% NaCl, when the acid concentration (regardless of type) was 0.2 to 0.3%. These results demonstrate the applicability of such model designs to include qualitative variables in investigations related to the development of growth-no growth limits.     

Microbiological and organoleptic characteristics of beef trim and ground beef treated with acetic acid, lactic acid, acidified sodium chlorite, or sterile water in a simulated commercial processing environment to reduce Escherichia coli O157:H7 and Salmonella

The objective of this study was to validate the effectiveness of acetic and lactic acids (2% and 5%), acidified sodium chlorite (1000 ppm), and sterile water in reducing Escherichia coli O157:H7 and Salmonella Typhimurium in inoculated beef trim in a simulated processing environment. Samples were collected to assess microbial characteristics at three processing points. Results from this study indicate that all treatments, including sterile water, reduced pathogen concentrations (P < 0.05) of both E. coli O157:H7 and Salmonella Typhimurium in ground beef up to 0.5 and 0.6 log by 24 h, respectively. In some cases, there were no significant differences between the antimicrobial treatments and the sterile water using this application method. Triangle sensory test results of non-inoculated beef indicated there were no differences (P < 0.05) in the means of correct responses between controls or antimicrobial treatments at 6 or 24 h. While interventions are important for beef trim, use of the interventions must be validated under industry conditions to ensure proper effectiveness.     

Use of response surface methodology in a fed-batch process for optimization of tricarboxylic acid cycle intermediates to achieve high levels of canthaxanthin from Dietzianatronolimnaea HS-1

In this work, we applied statistical experimental design to a fed-batch process for optimization of tricarboxylic acid cycle (TCA) intermediates in order to achieve high-level production of canthaxanthin from Dietzia natronolimnaea HS-1 cultured in beet molasses. A fractional factorial design (screening test) was first conducted on five TCA cycle intermediates. Out of the five TCA cycle intermediates investigated via screening tests, alfaketoglutarate, oxaloacetate and succinate were selected based on their statistically significant (P < 0.05) and positive effects on canthaxanthin production. These significant factors were optimized by means of response surface methodology (RSM) in order to achieve high-level production of canthaxanthin. The experimental results of the RSM were fitted with a second-order polynomial equation by means of a multiple regression technique to identify the relationship between canthaxanthin production and the three TCA cycle intermediates. By means of this statistical design under a fed-batch process, the optimum conditions required to achieve the highest level of canthaxanthin (13172 ± 25 μg l⁻¹) were determined as follows: alfaketoglutarate, 9.69 mM; oxaloacetate, 8.68 mM; succinate, 8.51 mM.     

Estimates of the efficiency of transfer of L‐histidine from blood to milk when it is the first‐limiting amino acid for secretion of milk protein in the dairy cow

The efficiency of transfer of L ‐histidine into milk protein was measured in two experiments in which L ‐histidine was infused intravenously into dairy cows eating a basal diet of grass silage and a cereal‐based supplement containing feather meal. Both experiments used Latin square designs, and infusion periods lasted 10 days. In Experiment 1, histidine was infused alone at doses of 3, 6 and 9 g day^?1. The output of milk protein increased up to the 6 g day^?1 dose but fell back to the basal level when 9 g day^?1 was infused. The efficiency of transfer was highest for the 6 g day^?1 dose, for which the value was 0.38. In Experiment 2 the same three histidine doses as in Experiment 1 were used, but this time the histidine was accompanied by 8 g L ‐methionine, 28 g L ‐lysine and 2.5 g L ‐tryptophan, to ensure that histidine remained first‐limiting over the whole dose range. The output of histidine in milk protein (Y) increased linearly with histidine dose (X) such that Y = 0.431 X + 0.070 r = 0.998; n = 4, indicating an efficiency of transfer of 0.43. © 2001 Society of Chemical Industry     

Use of novel PCR primers specific to the genes of staphylococcal enterotoxin G, H, I for the survey of Staphylococcus aureus strains isolated from food-poisoning cases and food samples in Taiwan

Data regarding the incidence of the newly found enterotoxigenic Staphylococcus aureus strains in food poisoning cases and in food samples were to date not available in Taiwan. In this study, PCR primers specific for the detection of SEG, H and I genes, i.e., seg, seh and sei, were used for the assay of 55 human isolates of S. aureus negative to the classical enterotoxins (SEA-->SEE) detection. These isolates were from the fecal specimens of the patients suffering from food poisoning outbreaks. Only eight strains were found to have the seg, seh and sei. The presence of other bacterial pathogens, such as Vibrio parahaemolyticus, Bacillus cereus and Salmonella spp. and perhaps, strains producing other new staphylococcal enterotoxins, in the fecal specimens of these patients, may account for these food poisoning cases. For 139 strains from food samples, such as frozen Chinese foods, Chinese sausages and lunch meals, sea strains accounted for the major portion and it seemed to be the most common SE type to coexist with seg, seh and sei. Only two strains had sec and none of them had seg, seh or sei. For strains without the classical SE genes, only 13 strains had seg, seh and/or sei. The above results imply that seg, seh and sei S. aureus strains play only a minor role in food-borne outbreaks in Taiwan.