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PubMed2004

Polychlorinated dibenzo-p-dioxins and dibenzofurans from food salt samples processed by heat treatment in Korea.

Eight samples of processed food salt collected from five plants in Korea were analyzed for 2,3,7,8-substituted polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) using liquid-liquid extraction, clean-up procedures, and high resolution gas chromatography-high resolution mass spectrometry. The study included the analyses of two kinds of salt product sample: bamboo-salt and parched salt. The levels of toxic PCDD/Fs found in the salt product samples were extremely low: the results revealed TEQ levels ranging between the sub pg TEQ/g and sub fg TEQ/g. The differences in the TEQ values of toxic PCDD/F were observed between the salt product samples, which were treated with different frequency of baking using four different fuels (firewood, pine wood, pine resin, and indirect heating by gas) at temperatures from 300 to 2000 degrees C. In bamboo-salt samples, the concentrations and TEQ values of toxic PCDD/Fs ranged between 0.57-66 pg/g and 5.7x10(-5)-0.64 pg TEQ/g, respectively. PCDD/Fs levels in bamboo-salts baked by firewood were found to be higher than those baked by pinewood or pine resin. In parched salt samples, the concentrations and TEQ values of toxic PCDD/Fs ranged between 0.97-3.7 pg/g and 0.0097-0.017 pg TEQ/g, respectively. The data was discussed regarding the concentration and the distribution pattern of congeners.

Benzofurans↗

PubMed1997

Mutagenic heterocyclic aromatic amines (HAAs) in 'processed food flavour' samples.

Eight samples of 'processed food flavours' (PFFs), chosen from five different categories, were analysed for their mutagenic activity using the Ames Salmonella assay, and also for the presence of eight heterocyclic aromatic amines (HAAs), namely 2-amino-3-(trideuteromethyl)imidazo[4,5-f]quinoline (IQ), 2-amino-3,8-dimethyl-imidazo[4,5-f]quinoline (MeIQ), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-amino-3,7,8-trimethylimidazo[4,5-f]quinoxaline (7,8-DiMeIQx), 2-amino-3,7,8-trimethylimidazo[4,5-f]quinoxaline (4,7,8-TriMeIQx), 2-amino-I-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) and 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2) using liquid chromatography and mass spectrometry (LC/MS). The isolation of HAAs was based on sequential liquid-liquid extraction procedures of samples at both acidic and basic pH values. The recoveries and the clean-up were monitored by introduction of quality control samples and by spiking with three tri-deuterated standards of HAAs. Although the results for the mutagenicity assay were comparable by testing less-purified and highly-purified extracts, the analysis for identification and quantification of HAAs by LC/MS required highly purified concentrates. Four samples had little or no mutagenic activity and these results were in agreement with their LC/MS results: they had no detectable levels (detection limits 1-3 ppb) of any of the HAAs monitored. The mutagenic activity of one sample was in complete agreement with the quantification of HAAs by LC/MS. Two samples produced strong mutagenic responses (3115 and 2664 revertants/g). In one sample, LC/MS analysis revealed the presence of 9.6 ppb IQ, whereas LC/MS of the other could not confirm the presence of any of the eight HAAs monitored. Two samples produced mild mutagenic activity (204 and 160 revertants/g), but relatively elevated concentrations of IQ (6.7 and 6.8 ng/g) by LC/MS. The extracts from all samples were tested for their modifying effects on mutagenicity of four HAAs. The discrepancy between the Ames test and the LC/MS analysis of some samples indicates several possibilities, such as the presence of some other HAAs, of their isomers or of other mutagens. In addition, the presence of mutagen modifiers (inhibitors or synergists) was observed in most samples. The results indicate that although chemical tests (e.g. LC/MS) can provide quantitative data for the HAAs monitored, the Ames mutagenicity test should also be conducted to determine the mutagenic activities of PFFs, in order to assess their health risk potential.

Amines↗

PubMed1995

Isolation, and virulence profiles, of Aeromonas hydrophila implicated in an outbreak of food poisoning in Sweden.

A case of food poisoning outbreak involving Aeromonas hydrophila is reported in this study. A group of 27 people consumed a typical Swedish food "landgång" which is a type of smörgåsbord containing shrimps with mayonnaise, liver paté, ham, sausage, and legume salad which was purchased from a food store. Twenty-two of the 27 persons became ill within 20-34 hr of consumption of the food and reported the symptoms ranging from severe acute diarrhea, abdominal pain, headache, fever and vomiting. One person also fainted. The symptoms lasted for a couple of days. Of the remaining 5 healthy persons who consumed the left-over food the next day, 2 became ill with similar symptoms. The bacteriological examination of left-over food samples resulted in the isolation of A. hydrophila from shrimps with mayonnaise, smoked sausage, liver paté and boiled ham. The total number of A. hydrophila in these foods were log 10(6) to log > 10(7) organisms per gram of food sample. A. hydrophila was however, not isolated from legume/mayonnaise salad samples. All the food samples tested showed low numbers of other expected food contaminating organisms such as coliforms at 37 C and 44 C, fecal streptococci, Staphylococcus aureus, fungi and yeast etc., while Bacillus cereus, Clostridium perfringens and Salmonella spp. were not detected in the food samples. Investigations of the virulence profiles of the A. hydrophila isolates showed their capacity to produce beta-hemolysin, cytotoxins, cytotonic toxins, enterotoxins, and adhesion to and invasion of human intestinal (Henle 407) cells in culture.

Acute Disease↗

PubMed1998

Comparison of the SimPlate coliform and Escherichia coli test with Petrifilm, three-tube MPN, and VRBA + MUG methods for enumerating coliforms and E. coli in food.

SimPlate for coliforms and Escherichia coli (CEc) is a new method for the detection and quantification of coliforms and E. coli in food. Internal validation of the method was carried out at IDEXX Laboratories (Westbrook, ME) with 180 food samples representing a variety of different food matrices and compared against three-tube MPN (most probable number), VRBA (violet red bile agar) + MUG, and Petrifilm (E. coli count) methods. SimPlate CEc was highly correlated with each of these methods for the quantification of coliform bacteria (r > or = 0.90). An insignificant number of food samples were found to contain E. coli; therefore, no meaningful correlation data could be generated. Four hundred forty-four additional food samples were tested at five collaborating laboratories for the presence of coliforms E. coli using SimPlate CEc and either VRBA + MUG or Petrifilm (E. coli count). Regression analysis of data from SimPlate for CEc versus Petrifilm E. coli count plates generated correlation coefficients (r) of at least 0.89 for total coliforms and at least 0.90 for generic E. coli. Correlation coefficients between SimPlate for CEc and VRBA + MUG data were at least 0.90 for coliforms and at least 0.86 for E. coli. SimPlate for CEc demonstrated better recovery of E. coli than Petrifilm when high populations of bacteria were present. E. coli was not detected in 20 or 50 (40%) raw milk samples tested by the Petrifilm method due to the presence of interfering coliform and noncoliform bacteria. It is concluded that SimPlate for CEc is a suitable alternative for determining numbers of coliform bacteria and E. coli in food.

Bacteriological Techniques↗

PubMed1993

Fluorescent detection-polymerase chain reaction (FD-PCR) assay on microwell plates as a screening test for salmonellas in foods.

This study evaluates a polymerase chain reaction assay coupled with a fluorescent detection in microwell plates for salmonellas in food samples. Chelex 100-extracted cultures and bulk and processed food samples were used as templates for a PCR assay in microwell plates, with a primer pair that amplifies a 206 bp segment of IS200. The PCR products were then denatured by heat and transferred to CovaLink NH plates (Nunc) to which capture oligonucleotides were covalently bound. Hybridization was performed for 1 h at 55 degrees C, the microwells were washed and an alkaline phosphatase-labelled probe, complementary of an internal sequence of the PCR product, was added. After stringent washes, 100 microliters of 1 mmol 1(-1) AttoPhos (JBL Scientific) was then added to the wells and the fluorescence measurement system (Millipore). The level of detection of the assay was as low as 1-10 cfu. A total of 172 food samples were tested, both by culture and FD-PCR. Of these 53 were culture positive and 119 culture negative. The sensitivity of the FD-PCR assay was 100% and the specificity was 90.1%. Positive and negative predictive values were 82.8 and 100%, respectively. Based on the results obtained in this study it appears that the FD-PCR assay described here can be useful to screen a large number of food samples for contamination by salmonellas.

Alkaline Phosphatase↗

PubMed2000

Development of methods to detect "Norwalk-like viruses" (NLVs) and hepatitis A virus in delicatessen foods: application to a food-borne NLV outbreak.

"Norwalk-like viruses" (NLVs) and hepatitis A virus (HAV) are the most common causes of virus-mediated food-borne illness. Epidemiological investigations of outbreaks associated with these viruses have been hindered by the lack of available methods for the detection of NLVs and HAV in foodstuffs. Although reverse transcription (RT)-PCR methods have been useful in detecting NLVs and HAV in bivalve mollusks implicated in outbreaks, to date such methods have not been available for other foods. To address this need, we developed a method to detect NLVs and HAV recovered from food samples. The method involves washing of food samples with a guanidinium-phenol-based reagent, extraction with chloroform, and precipitation in isopropanol. Recovered viral RNA is amplified with HAV- or NLV-specific primers in RT-PCRs, using a viral RNA internal standard control to identify potential sample inhibition. By this method, 10 to 100 PCR units (estimated to be equivalent to 10(2) to 10(3) viral genome copies) of HAV and Norwalk virus seeded onto ham, turkey, and roast beef were detected. The method was applied to food samples implicated in an NLV-associated outbreak at a university cafeteria. Sliced deli ham was positive for a genogroup II NLV as determined by using both polymerase- and capsid-specific primers and probes. Sequence analysis of the PCR-amplified capsid region of the genome indicated that the sequence was identical to the sequence from virus detected in the stools of ill students. The developed method is rapid, simple, and efficient.

Animals↗

PubMed1999

Detection of Salmonella spp. in food by a rapid PCR-hybridization procedure.

A rapid and sensitive PCR-hybridization procedure for detection of Salmonella serovars in food samples was developed. This method is based on three subsequent steps: (1) extraction of nucleic acids from a 2 ml aliquot of the pre-enrichment medium used for the conventional culture method after 6 h of incubation at 37 degrees C; (2) amplification with primers selected from the sequences of invE and invA genes; (3) Southern blot and hybridization with a biotin labeled oligonucleotide probe. The entire procedure requires 30 h. The PCR-hybridization assay was able to detect as little as 50 fg of purified chromosomal DNA of S. typhimurium and 0.2 cfu g-1 of an artificially contaminated food sample. Of 245 food samples analyzed by culture and PCR-hybridization, 20 were positive by both methods and 16 were positive by PCR-hybridization only. None of the 209 PCR-negative samples tested positive by culture. The sensitivity, specificity, alpha and beta error values of the results of the PCR-hybridization procedure, compared with those of culture, were 100, 92.9, 0 and 7.1%, respectively. These results indicate that a short pre-enrichment and PCR-hybridization could be used as a screening test for the detection of Salmonella in food samples.

Animal Feed↗

PubMed2006

Listeria monocytogenes food monitoring data and incidence of human listeriosis in Hungary, 2004.

Food samples were monitored for contamination with Listeria monocytogenes, and the incidence of human listeriosis was evaluated according to the data obtained in Hungary in the year 2004. Of the food samples tested, the bacterium was most often detectable in milk and dairy products, as 72.1% of all L. monocytogenes strains were isolated from these samples. The food samples most commonly yielded strains of serotype 1/2a (45.1%) and 4b (27.0%). In 2004, 3 perinatal and 14 nonperinatal human listeriosis cases were diagnosed in Hungary. These disease cases were most often caused by strains belonging to serotype 4b (52.8%) and serotype 1/2a (23.5%). On the basis of the antibiotic sensitivity test results of strains isolated from the disease cases, penicillin and aminoglycoside antibiotics or a combination thereof were found to be effective.

Adult↗

PubMed1979

Mass spectrometric identification and gas-liquid chromatographic determination of 2-chloroethyl esters of fatty acids in spices and foods.

The 2-chloroethyl esters of 5 fatty acids have been identified in spice and food samples by gas-liquid chromatography-mass spectrometry (GLC/MS). Twenty-four spice samples were analyzed for the 2-chloroethyl esters of fatty acids by AOAC official multiple residues pesticide procedure using GLC with microcoulometric detection. The esters of capric, lauric, myristic, palmitic, and linoleic acids have been identified at levels up to 1400 ppm. 2-Chloroethyl linoleate was the most abundant ester in all samples. Several foods analyzed by the same procedures showed levels of 2-chloroethyl linoleate as high as 35 ppm. Recoveries from fortified samples ranged from 84 to 98% for the various esters. A method using an acid-catalyzed esterification reaction was developed to rapidly determine the fatty acid content of these spices. GLC analysis with microcoulometric detection was used. Recoveries from fortified samples ranged from 92 to 110%. After 2 spice samples found to be free of 2-chloroethyl esters were fumigated with ethylene oxide, the level of 2-chloroethyl linoleate reached 77 ppm. All levels of 2-chloroethyl esters were confirmed by GLC/MS.

Chromatography, Gas↗

PubMed2000

Detection of freeze-injured Escherichia coli O157:H7 cells from foods by resuscitation prior to selective enrichment.

We tried to detect Escherichia coli O157:H7 in food samples artificially contaminated with freeze-injured E. coli O157:H7 using an enrichment method with modified EC broth supplemented with novobiocin (mEC + n). When the samples were cultured for enrichment immediately after inoculation of freeze-injured cells, E. coli O157:H7 was not detected in 13 out of 18 samples. However, allowing the food samples to stand for 3 h at room temperature prior to enrichment in mEC + n remarkably improved recovery of E. coli O157:H7 except for some acidic foods. E. coli O157:H7 was detected in the acidic foods by introducing a resuscitation step of 3-h of incubation in a non-selective broth at room temperature prior to enrichment with mEC + n.

Anti-Bacterial Agents↗

PubMed2005

[The characteristic of Staphylococcus aureus strains isolated from food's samples].

The study was carried out of 22 isolates of S. aureus isolated from 7 different incriminated food's samples from foodborne-disease outbreaks. The possibility of these isolates to producing of enterotoxins by commercial test SET-RPLA (Oxoid) was tested. The genotyping of these isolates was done by pulse-filed gel electrophoresis, acc. to Pfaller in own modification. On the basis of the DNA restriction patterns of the 22 isolates--5 strains were singled out, one of these strains--strain V (isolat nr 7) was not relationship to others. It was found that this strain V was one enterotoxin produced. Additionaly, all tested strains, in spite of the strain nr V, were isolated from the 2 or 3 samples of different kinds of foods. In the present study it has been shown too, that several similar colonies should be isolated for farther studies to assess microbiological contamination of the food products properly.

Enterotoxins↗

PubMed2005

Characterization of diarrheagenic Escherichia coli isolated from food in Khon Kaen, Thailand.

Four categories of 186 ready-to-eat food samples in Khon Kaen municipality, Thailand, were collected and investigated for fecal contamination by enumeration of Escherichia coli using the most probable number (MPN) method. Then, the E. coli isolates were presumptively identified as diarrheagenic E. coil by agglutinating with polyvalent O-antisera and monovalent O-antisera commonly found in diarrheagenic strains and were subsequently investigated for the presence of the recognized virulence genes for enteroaggregative (EAEC), enteroinvasive (EIEC), enteropathogenic (EPEC), enterotoxigenic (ETEC), and shiga toxin-producing E. coli (STEC or EHEC) by multiplex PCR assays. All E, coli isolates were examined for antimicrobial susceptibilities by the agar disc diffusion method, and the results were compared with those obtained from clinical samples. The percentage of each type of food with E. coli, including no heat food, low heat food, high heat food, and fruit juices and beverages, was higher than accepted standards at 60.4, 46.5, 38.6 and 20%, respectively. Of 140 E. coli isolates obtained from food samples, 11 isolates (7.9%) agglutinated with 6 monovalent O-antisera, including one isolate each of O6, O8, O114 and O159, two isolates of O1, and five isolates of O157. None of the 11 isolates harbored the virulence genes for EPEC, ETEC, EAEC, EIEC and STEC. Although O157 E. coli isolates were found, the most frequent, E. coli O157:H7, was not found in this study. The astA gene, however, was found in 1 E. coli isolate that showed weakly positive agglutination against the polyvalent antisera. Approximately 50% of the 140 E. coli isolates were resistance to at least one antimicrobial agent. The resistant strains showed high resistance to tetracycline (43%), co-trimoxazole (36%), ampicillin (26%) and chloramphenicol (23%), respectively. The resistance of E. coli was high for nearly all antimicrobial agents, particularly ampicillin (76%), tetracycline (70%), co-trimoxazole (69%) and nalidixic acid (44%). The results show that nearly half of the ready-to-eat food samples evaluated in Khon Kaen Municipality had levels of E. coli higher than acceptable standards. Of the diarrheagenic E. coli classified by serogroup, almost none of the isolates had virulence genes. These results indicate the disadvantage of relying on serogrouping alone for the recognition of diarrheagenic E. coli. E. coli isolated from food may not be an enteropathogenic strain. We also found that E. coli antimicrobial resistant strains are widespread in both food and humans.

Animals↗

PubMed2006

Multiplex real-time PCR detection of heat-labile and heat-stable toxin genes in enterotoxigenic Escherichia coli.

A multiplex real-time PCR method was developed for detection of heat-labile and heat-stable toxin genes in enterotoxigenic Escherichia coli. Approximately 10 CFU per reaction mixture could be detected in rinsates from produce samples. Several foods representative of varieties previously shown to have caused enterotoxigenic E. coli outbreaks were spiked and enriched for 4 or 6 h. Both heat-labile and heat-stable toxin genes could be detected in the foods tested, with the exception of hot sauce, with threshold cycle values ranging from 25.2 to 41.1. A procedure using membrane filtration which would allow enumeration of the enterotoxigenic E. coli population in a food sample in less than 28 h by real-time PCR analysis of colonies picked from media highly selective for E. coli was also developed.

Amino Acid Sequence↗

PubMed1992

16S rRNA-based probes and polymerase chain reaction method to detect Listeria monocytogenes cells added to foods.

A rapid polymerase chain reaction (PCR) method was developed for detection of Listeria monocytogenes in foods. This method used a pair of primers based on a unique region in the 16S rRNA sequence of L. monocytogenes, which were previously reported by us to yield a specific nucleic acid probe. Our method included use of a shorter denaturing time, a shorter annealing time, a rapid transition, and an increase in the number of cycles, resulting in good sensitivity. Just 3 h for PCR plus 1 h for electrophoresis was required. Additional time for DNA isolation and DNA hybridization was not needed. This method detected as few as 2 to 20 CFU of L. monocytogenes in pure cultures and as few as 4 to 40 CFU of L. monocytogenes in inoculated (10(8) CFU), diluted food samples. Seven of eight foods, including four poultry products, gave positive results. Only one food sample, soft cheese, gave interference. An internal probe hybridization test was used to confirm that the PCR products were from L. monocytogenes. A specificity test indicated that this PCR method was positive for all 13 strains of L. monocytogenes tested but negative for the other 6 species of Listeria, including 6 strains of L. innocua, and negative for 17 other gram-positive and gram-negative bacteria tested.

Base Sequence↗

PubMed2005

[Molecular analysis of Salmonella enteritidis by pulsed-field gel electrophoresis].

OBJECTIVE: To study the relationship between different strains of Salmonella enteritidis, a molecular epidemiologic analysis method-pulsed-field gel electrophoresis was established and Salmonella enteritidis isolated from food samples were tested. METHODS: Food samples were collected from Henan province, Heilongjiang province and other provinces in 1991 - 2001. Strains were isolated and single colony of each Salmonella enteritidis strain from an agar plate was picked and enriched, then the total DNA was extracted in turn with lysozyme solution, proteinase K solution and TE buffer. The agarose pulgs were digested with restriction enzyme XbaI overnight at 37 degrees C, followed by pulsed-field gel electrophoresis (PFGE) with a CHEF Mapper system in 0.5 x TBE buffer with recirculation at 14 degrees C . DNA macrorestriction fragments were resolved on 1% agarose gels, pulse time were ramped from 5s to 45s during a 26h run at 6V/cm. A lambda ladder pulsed-field gel maker was used as a size standard. RESULTS: 30 Salmonella enteritidis strains were identified in chicken, beef, cooked meat and vegetables. 18 strains were isolated from henna province. More of the stains (22 strains) were isolated in 2001. Two distinct DNA fragment profiles, PFGE types A and B, were observed, representing 63.3% and 36.7% of the isolates respectively. Type A could be subdivided further into two subtypes. Subtype A1 represented the majority (78.9%) of type A strains and differed from subtype A2 in that the latter lacked a band in the 50kb region. There was no correlation between the strains profiles and their isolation time or region. However, all the strains isolated from the chicken were totally type A1, and showed certain epidemiological trend which can be studied in the future. CONCLUSION: The pulsed-field gel electrophoresis is a good method to study the molecular epidemiological relationship between the different Salmonella enteritidis srtains.

Bacterial Typing Techniques↗

PubMed2004

Electrochemical detection of ascorbic acid (vitamin C) using a glassy carbon electrode.

A method for the detection of ascorbic acid (AA) was developed using square wave anodic stripping voltammetric (SWASV) analysis with a glassy carbon electrode (GCE). The results indicated a sensitive oxidation peak current of AA on the GCE. A linear curve was obtained within a concentration range of 1-130.0 microg/L with a preconcentration time of 270 s. The relative standard deviation of 30.0 microg/L observed was 0.1016% (n = 12) under optimum conditions. The low detection limit (S/N) was pegged at 0.30 microg/L. Results showed that the method developed can be used to assay biological and pharmaceutical samples, and food samples, as well as other materials requiring AA analyses.

Antioxidants↗

PubMed2003

Benefits of high resolution IC-ICP-MS for the routine analysis of inorganic and organic arsenic species in food products of marine and terrestrial origin.

A recently developed and validated method for simultaneous determination of 17 inorganic and organic arsenic compounds in marine biota has been successfully applied to routine analysis of different food products, including fish, shellfish, edible algae, rice, and other types of grain. During one year, approximately 250 food samples were analyzed, mostly fish and rice. Long-term stability and robustness of the system was observed and reproducible results for certified reference materials were ensured by means of control charts. The separation was performed by ion-pair chromatography on an anion-exchange column to separate anionic, neutral, and cationic arsenic species in one chromatographic run. Hyphenation to ICP-MS allowed element-specific and sensitive detection of the different arsenic species with a detection limit as low as 8 ng As L(-1 )in the sample extract, which is equivalent to 2 ng As g(-1) in the original sample. Special emphasis was laid on the analysis of marine algae and rice samples. These food types can contain elevated levels of the very toxic inorganic arsenic species (up to 90% in rice) and therefore are the focus of interest in the food industry. In marine algae, inorganic arsenic was mainly present as arsenate whereas in rice arsenite predominated.

Arsenicals↗

PubMed1979

Infant foods as a potential source of diarrhoeal illness in rural West Africa.

It is common practice in rural areas of The Gambia to prepare infant foods in quantities which are sufficient to meet the needs of the day rather than one meal. These are then stored at ambient temperatures for periods up to 12 hours for feeding to the child on demand. The total viable counts and levels of Bacillus cereus, Clostridium welchii, Staphylococcus aureus and Escherichia coli were determined in 294 infant foods samples from nought to eight hours after preparation. The presence of Salmonella was determined in 10 g samples of food. In the first hour after preparation the proportion of foods dangerously contaminated was high during the rainsy season, significantly more so than during the dry season. Foods not consumed fresh were very often hazardous and almost always so after 8 hours. This problem may be a causal factor in weanling diarrhoea which also shows marked seasonal variation in prevalence.

Diarrhea, Infantile↗