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Illness outbreak associated with Escherichia coli O157:H7 in Genoa salami. E. coli O157:H7 Working Group.

BACKGROUND: An outbreak of Escherichia coli O157:H7 infection was identified in the spring of 1998, with a 7-fold increase in the number of laboratory-confirmed E. coli O157:H7 cases in southern Ontario. This prompted an intensive investigation by local, provincial and federal public health officials. METHODS: Case interviews of 25 people from southern Ontario were conducted using a broad food history and environmental exposure survey. Laboratory investigations involved both case and food sampling. Specimens of foods sold locally and reportedly consumed by those affected were tested. Common suppliers of suspected foods were identified by cross-referencing suppliers' lists with stores frequented by those who fell ill. A case-control study involving 25 cases and 49 age-matched controls was conducted. This was followed by a comprehensive environmental investigation of the meat processing plant identified as the source of the E. coli. RESULTS: Thirty-nine outbreak-related cases occurred between April 3 and June 2, 1998. Of the 36 case specimens tested all were positive for E. coli O157:H7. The case-control study identified Genoa salami as the most probable (odds ratio 8 [confidence interval 2-35]) source of the outbreak. Samples of Genoa salami produced by the most commonly identified supplier later tested positive for E. coli O157:H7, and the pathogen matched the same pulsed-field gel electrophoresis pattern and phage type of the case specimens. INTERPRETATION: Our investigation, which led to a national recall of the brand of dry fermented Genoa salami identified as the source of the outbreak, supports an adherence to stringent manufacturing requirements for fermented meat products. A review of the Canadian standards for fermented meat processing and the effectiveness of their implementation is warranted.

Adolescent↗

Occurrence of non-O157 shiga toxin-producing Escherichia coli in ready-to-eat food from supermarkets in Argentina.

Between June 2000 and December 2001, 500 food samples were collected from supermarkets and shops selling ready-to-eat food in Rosario, Argentina, and examined for Escherichia coli. Forty-nine E. coli isolates from food samples were further characterized for virulence genes by multiplex polymerase chain reaction (PCR) targeting the stx1, stx2, stx2e, eaeA, CNF1, CNF2, Einv, LTI, STI, and STII genes in four groups. Out of 49 E. coli isolates screened by multiplex PCR, only 10 possessed Shiga toxin genes, stx1 and stx2 genes and none possessed the other genes. The Shiga toxin positive E. coli strains (STEC) were isolated from soft, cottage cheeses, chicken with sauce and vegetables mayonase. These E. coli isolates were serogrouped and belonged to O18 (two strains), O8, O57w, O79, O44, and O128; three strains were untypeable. Pulsed-field gel electrophoresis (PFGE) with XbaI generated a unique profile for each, having 10-15 bands ranging from 50 to 500 kb, except that strain ARG 20 generated small bands and was partly degraded. These strains are potential foodborne pathogens and their presence in ready-to-eat food illustrates the need to keep a careful watch for the source of pathogens and then develop methods to control them.

Argentina↗

Rapid and sensitive immunomagnetic-electrochemiluminescent detection of staphyloccocal enterotoxin B.

Sensitive, rapid and reproducible detection of staphyloccocal enterotoxin B (SEB) in a range of different biological matrices was achieved using the ORIGEN((R)) Immunoassay System (Igen, Inc). The homologous immunoassay format consisted of a double antibody sandwich in which a biotinylated capture antibody, pre-bound to streptavidin-coated paramagnetic beads, was used to bind antigen from test samples. A detector antibody, labeled with ruthenium (II) tris-bipyridal chelate, was added and, when bound to the bead immunocomplex, generated light in the presence of an excess of tripropylamine. The light was detected and measured by the ORIGEN analyzer. The sensitivity of this assay was 1 pg of enterotoxin per ml of serum, urine, tissue, or buffer and was highly reproducible. Concentration curves generated from SEB standards produced consistently wide linear ranges (0.1-100 ng/ml), making quantitation possible with only two dilutions of sample (undiluted and 1:1000). The assay used 50 microl of sample per test and required a 30 min incubation period in addition to a 1 min per tube reading time (50 tubes maximum). This assay was significantly better in terms of sensitivity, linear range, and assay time than the standard microplate enzyme-linked immunosorbent assay and should permit early SEB detection in clinical samples, food, and environmental samples.

Animals↗

The effects of domestic cooking on the levels of 3-monochloropropanediol in foods.

The results are reported of a study to determine the effect of domestic cooking procedures on the level of 3-monochloropropane-1,2-diol (3-MCPD) in selected foods. Samples of 23 foods comprising stock cubes, gravies, a cake mix, batters, breads, cheese and meats were subjected to a range of cooking procedures including grilling, toasting and microwaving. The resulting levels of 3-MCPD were compared with those present in the foods before cooking. Grilling and toasting produced substantial increases in the 3-MCPD content of bread, forming up to 0.3 mg/kg, and of most cheeses, resulting in levels of up to about 0.1 mg/kg. Microwave cooking produced elevated 3-MCPD levels in some cheeses. Frying laboratory-produced batters increased 3-MCPD levels to about 0.1 mg/kg whereas a retail batter contained no detectable 3-MCPD when fried. The remaining foods showed little or no discernible increase on cooking.

Bread↗

Acrylamide in Asian foods in Hong Kong.

About 400 food samples, mainly Asian foods available in Hong Kong, were tested for acrylamide by an LC-MS/MS method using [1, 2, 3-(13)C(3)]-acrylamide as surrogate. The acrylamide levels in the more commonly consumed food items in the food groups such as rice and rice products, noodles, bakery and batter-based products, were generally less than 60 microg kg(-1). Higher levels were found in the food groups such as biscuit-related products and crisps. The highest levels were detected in potato crisps (1500-1700 microg kg(-1)). Lower levels were found in rye flour-based crisps (440 microg kg(-1)), followed by corn-based (65 to 230 microg kg(-1)) and wheat flour-based crisps (61-200 microg kg(-1)), and then rice flour-based crisps (15-42 microg kg(-1)). The acrylamide formation during deep frying of a wheat flour-based product, Chinese fried fritter, was studied. Deep-frying at 170 degrees C resulted in gentle but steady rise in acrylamide content. A steep rise for frying at 210 degrees C was recorded. The moisture content of the product decreased with frying time, but the fat content increased. It is proposed that the reaction for the formation of acrylamide was initiated on the surface and then penetrated into the interior of the food matrix by heat transfer via radiation/conduction and diffusion of hot oil.

Acrylamide↗

Future directions in food composition studies.

Food composition studies are essential for many purposes in nutrition, food science, and dietetics. This article reviews uses of food composition information in food labeling, nutrition education, dietary surveys, epidemiological studies, and dietary intervention trials. Research is need to minimize the causes of variation between actual and measured diets. This involves minimizing errors in food sampling, chemical analysis, food composition table construction, nutrient data bases for food groups, and in computations using nutrient analysis systems. Critical knowledge relative to food composition necessary for food science experts, nutrition professionals and other professionals is discussed. Improving the reporting of food composition data will also require more attention by editors of journals: if they demand more documentation, they will get it.

Diet↗

Use of the affinity/HPLC method for quantitative estimation of folic acid in enriched cereal-grain products.

In 1998, the United States introduced mandatory fortification of enriched cereal-grain products with folic acid to reduce the incidence of neural tube defects. As a consequence, substantial amounts of folic acid, the synthetic form of folate, were added to the American diet, and the ability to assess folic acid intake took on greater importance. The purpose of the current study was to separate and quantify folic acid and 5-methyltetrahydrofolate, the most prominent naturally occurring folate in fortified foods, with a reliable and robust method. Folates were heat-extracted from food samples. A trienzyme treatment (alpha-amylase, rat plasma conjugase, and protease) was applied to the extracts followed by purification by affinity chromatography. Folic acid and 5-methyltetrahydrofolate were separated and quantified by reversed-phase HPLC with fluorescence and UV detection. A gradient elution with phosphate buffer and acetonitrile was used to separate the different forms of folates. The method gave a linear response in a range of 0.1-3 mumol/L and 0.0125-0.25 mumol/L for folic acid and 5-methyltetrahydrofolate, respectively. These ranges were similar to the expected levels in the samples. The CV of the peak areas of folic acid and 5-methyltetrahydrofolate for 5 commercial wheat flour samples extracted and run separately on the same day was 2.0 and 5.7% and, run over 5 consecutive days, was 7.2 and 7.3%, respectively. Total folate values in 45 samples of fortified food measured by HPLC and by the traditional microbiological assay demonstrated a high correlation (r(2) = 0.986).

Chromatography, Affinity↗

Enterotoxigenicity and genetic relatedness of Clostridium perfringens isolates from retail foods in the United States.

Clostridium perfringens is a leading cause of bacterial food-borne illness in countries where consumption of meat and poultry is high. For example, each year in the United States, this organism is the second or third most common cause of confirmed cases of food-borne illness. Surveys of the incidence of this organism in retail foods were done in the 1960s without regard to whether isolates were enterotoxigenic. It is now known that not all strains of this organism possess the enterotoxin gene responsible for illness. We examined the incidence of this organism in 131 food samples from retail food stores in an area of the northeastern United States. Forty isolates were obtained by using the iron milk method at 45 degrees C, with confirmation by use of motility nitrate and lactose gelatin media. The presence of the C. perfringens enterotoxin (cpe) and alpha toxin (cpa) genes was determined by PCR using previously published primer sequences. All isolates possessed cpa. None of the isolates were identified as carrying the cpe gene by this method or by another method using a digoxigenin-labeled gene probe. Consistent with these results, none of the sporulating-cell extracts contained enterotoxin as determined by reverse passive latex hemagglutination. Pulsed-field gel electrophoresis was used to determine the genetic relatedness of the isolates. About 5% of the isolates were considered to be closely related (2- to 3-band difference). The others were considered to be unrelated to one another. The results demonstrate the rarity of cpe(+) strains in retail foods and the genetic diversity among nonoutbreak strains.

Animals↗

[Evaluation of the incidence and toxicity of samples of Bacillus cereus in various classes of foods sold and consumed in the State of Rio de Janeiro].

One hundred and fourteen strains of Bacillus cereus were isolated during presumptive plate-counts from 18 groups of industrialized, non-industrialized, crude or cooked food, belonging to 10 separate classes. Specific presumptive counts ranged from 10(2) to 6 X 10(3)/g or ml. Among these isolates, 13 strains were derived from 3 outbreaks of food poisoning (involving a minimum of 57 people), as determined by the assayed bacteriological quality of the ingested foods. As an adopted procedure to correlate toxicity and ability to promote illness in man, culture fluids of all strains were assayed to determine their ability to increase vascular permeability (APC) to cause necrosis in rabbits skin and to kill albino mice. APC was positive in 86.85% of the 114 strains, death of albino mice occurred in 65.79% and a combination of APC and death was observed in 59.65%. APC plus necrosis, or only necrosis, occurred with 34.21% of the culture fluids. Death, APC and death with or without necrosis, were demonstrated in the strains implicated with illness. This confirmed the known individuality of action exhibited by certain B. cereus food-borne toxigenic factors. The low presumptive counts of this bacterium in the order of 10(2)-10(3)/g or ml found in food, implicated or not with illness, suggests that the recommended number of B. cereus per g or ml of food sample should be reevaluated in our country. Furthermore, a wider range of food should be brought under bacteriological sanitary control for this species.

Animals↗

Trans fatty acid isomers in human health and in the food industry.

Trans fatty acids are unsaturated fatty acids with at least one double bond in the trans configuration. These fatty acids occur naturally in dairy and other natural fats and in some plants. However, industrial hydrogenation of vegetable or marine oils is largely the main source of trans fatty acids in our diet. The metabolic effect of trans isomers are today a matter of controversy generating diverse extreme positions in light of biochemical, nutritional, and epidemiological studies. Trans fatty acids also have been implicated in the etiology of various metabolic and functional disorders, but the main concern about its health effects arose because the structural similarity of these isomers to saturated fatty acids, the lack of specific metabolic functions, and its competition with essential fatty acids. The ingestion of trans fatty acids increases low density lipoprotein (LDL) to a degree similar to that of saturated fats, but it also reduces high density lipoproteins (HDL), therefore trans isomers are considered more atherogenic than saturated fatty acids. Trans isomers increase lipoprotein(a), a non-dietary-related risk of atherogenesis, to levels higher than the corresponding chain-length saturated fatty acid. There is little evidence that trans fatty acids are related to cancer risk at any of the major cancer sites. Considerable improvement has been obtained with respect to the metabolic effect of trans fatty acids due the development of analytical procedures to evaluate the different isomers in both biological and food samples. The oleochemical food industries have developed several strategies to reduce the trans content of hydrogenated oils, and now margarine and other hydrogenated-derived products containing low trans or virtually zero trans are available and can be obtained in the retail market. The present review provides an outline of the present status of trans fatty acids including origin, analytical procedures, estimated ingestion, metabolic effects, efforts to reduce trans isomers in our diet, and considerations for future prospects on trans isomers.

Dietary Fats↗

Incidence and identification of mesophilic Aeromonas spp. from retail foods.

Sixty-eight food samples were examined for the presence of mesophilic Aeromonas species both qualitatively and quantitatively. Aeromonads were isolated from 26% of the vegetable samples, 70% of the meat and poultry samples and 72% of the fish and shrimps. Numbers of motile aeromonads present in the food samples varied from <10(2) cfu g(-1) to >10(5) cfu g(-1). GLC analysis of FAMEs was used to identify a selection of presumptive Aeromonas colonies to fenospecies or genomic species level. Aeromonas strains belonging to the Aer. caviae complex, which also includes the potentially pathogenic genospecies HG4, were mostly isolated from vegetables but were also found in meat, poultry and fish. In addition, three strains of the virulent taxon Aer. veronii biovar sobria HG8 were isolated from poultry and minced meat. All members of the Aer. hydrophila complex, predominant in the fish, meat and poultry samples, were classified in the non-virulent taxon HG3. Although the significance of Aeromonas in foods remains undefined, the isolation of Aeromonas HG4 and HG8 strains from a variety of retail foods may indicate that these products can act as possible vehicles for the dissemination of food-borne Aeromonas gastroenteritis.

Aeromonas↗

Phage types, virulence genes and PFGE profiles of Shiga toxin-producing Escherichia coli O157:H7 isolated from raw beef, soft cheese and vegetables in Lima (Peru).

The present study was conducted in Lima Metropolitana to evaluate the prevalence of Shiga toxin-producing Escherichia coli (STEC) O157:H7 in raw beef, raw ground beef, soft cheese and fresh vegetables, sampled at different markets in the city. Between October 2000 and February 2001, 407 food samples were collected from different markets in the 42 districts of Lima Metropolitana. Samples were assayed for E. coli O157 by selective enrichment in modified Tryptic Soy Broth containing novobiocin, followed by immunomagnetic separation (IMS) and plating onto sorbitol MacConkey agar supplemented with cefixime and potassium tellurite. Fifty (12.3%) of 407 food samples resulted positive for E. coli O157 isolation (23 of 102 ground beef; 15 of 102 beef meat; eight of 102 soft cheese and four of 101 fresh vegetables). Thirty-five E. coli O157 isolates were further analysed for the presence of virulence genes. All 35 were positive by PCR for O157 rfbE, fliCh7, eae-gamma1 and ehxA genes. In addition, genes encoding Shiga toxins were detected in 33 of 35 isolates, five isolates (14%) encoded stx(1), stx(2), and 28 (80%) stx2 only. The isolates were of seven different phage types (PT4, PT8, PT14, PT21, PT34, PT54, and PT87) with three phage types accounting for 80% of isolates: PT4 (15 isolates), PT14 (8 isolates), and PT21 (5 isolates). Interestingly, the majority (31 of 35; 89%) of E. coli O157:H7 isolates characterized in this study belonged mainly to the phage types previously found in STEC O157:H7 strains associated with severe human disease in Europe and Canada. Pulsed-field gel electrophoresis (PFGE) of 32 isolates revealed 14 XbaI-PFGE groups (I to XIV) of similarity >85%, with 23 (72%) isolates grouped in five clusters. Some isolates from different districts presented a high clonal relatedness. Thus, PFGE group VIII clustered eleven strains from nine different districts. The broad range of PFGE subtypes found in this study demonstrates the natural occurrence of many genetic variants among STEC O157:H7 spread in Lima.

Animals↗

[Listeria spp. in food of animal origin].

Different food samples of animal origin were analyzed for Listeria spp. Five L. innocua strains, one L. monocytogenes strain and one L. welshimeri strain were obtained from 208 samples of raw milk. The strains were typified by biochemical and serologic tests. The shortened enrichment method was chosen for isolations; Palcam and Oxford agar also permitted the growth of the seven strains. L. monocytogenes was recovered from milk of an animal with subclinical mastitis. No Listeria strains were isolated from pasteurized milk, chocolate milk or cheese samples. One L. welshimeri strain was detected in ice cream. In the case of meat food samples, the employment of two-step enrichment methods facilitated the detection of Listeria spp. A prevalence of L. ivanovii was observed in 2.5% of these samples.

Animals↗

Detection of Campylobacter jejuni and Campylobacter coli in foods by enrichment culture and polymerase chain reaction enzyme-linked immunosorbent assay.

A polymerase chain reaction (PCR) assay based on a solution hybridization format with colorimetric end-point detection (PCR ELISA) was investigated for the specific detection of Campylobacter jejuni and Campylobacter coli in food samples following enrichment culture. One hundred fifteen samples of raw meat and offal (poultry, porcine, ovine, and bovine), raw shellfish, and artificially contaminated milk were enriched in blood-free Campylobacter Enrichment Broth for 48 h. Enrichment cultures were subcultured to Campylobacter blood-free selective agar plates, and presumptive isolates were identified by phenotypic methods. DNA was extracted from 1-ml aliquots of the enrichment cultures using a rapid extraction method, and the DNA was used as the template in a PCR ELISA. A comparison of the PCR ELISA with the enrichment culture and subculture to selective agar method showed that the results of 112 of the 115 samples tested were in agreement by both methods. Seventy-one of the various food samples were positive in the PCR ELISA, and 70 samples were positive by culture. The PCR ELISA had a sensitivity of 99% and a specificity of 96%, with a positive predictive value of 97% and a negative predictive value of 98%. The PCR ELISA is a rapid, sensitive, and specific method for the detection of C. jejuni and C. coli in foods following enrichment culture and significantly reduces the time required for their detection.

Animals↗

[Survey of the presence of bacterial pathogens in foods sold at retail stores in the city of Cassino].

The presence of bacterial food pathogens was evaluated in 154 food samples collected from supermarkets and butchers in the city of Cassino (South-Central Italy). Food pathogens were identified in 17.5% of the total food samples. In the raw meat samples, 24.6% tested positives for Listeria monocytogenes, 4.3% for Salmonella and 2.9% for Escherichia coli O157. Y. enterocolitica, only investigated in pork meat, was identified in 7.4% of the samples. In poultry, L. monocytogenes was identified in 55% of the samples.

Animals↗

Multiresidue analysis of pesticides in vegetables and fruits using a high capacity absorbent polymer for water.

A single extraction and a single clean-up procedure was developed for multi-residue analysis of pesticides in non-fatty vegetables and fruits. The method involves the use of a high capacity absorbent polymer for water as a drying agent in extraction from wet food samples and of a graphitized carbon column for clean-up. A homogeneously chopped food sample (20 g) and polymer (3 g) were mixed to absorb water from the sample and then 10 min later the mixture was vigorously extracted with ethyl acetate (100 ml). The extract (50 ml), separated by filtration, was loaded on a graphitized carbon column without concentration. Additional ethyl acetate (50 ml) was also eluted and both eluates were concentrated to 5 ml for analysis. The procedure for sample preparation was completed within 2 h. In a recovery test, 107 pesticides were spiked and average recoveries were more than 80% from asparagus, orange, potato and strawberry. Most pesticides were recovered in the range 70-120% with usually less than a 10% RSD for six experiments. The results indicated that a single extraction with ethyl acetate in the presence of polymer can be applied to the monitoring of pesticide residues in foods.

Food Contamination↗

Hazard analysis and critical control points of weaning foods.

Hazards and critical control points were identified during the preparation, feeding and storage of weaning foods fed to the children (6-24 months) belonging to low income group (LIG) families using survey methods, spot and indepth observations and microbial analysis. High microbial counts were obtained for the weaning food samples procured from the families that were rated as poor for both personal hygiene and environmental sanitation. Food samples that were held at ambient temperature (25-35 degrees C) showed high counts of coliforms, AMCC, yeast and mold, Staphylococcus aureus and Bacillus cereus. Spice mixtures and mop samples also revealed high counts for all these organisms. Coliforms were not isolated from the water samples of all the 10 households. However, a high count of Bacillus cereus (10(3-5)/ml) was observed in water samples of two selected households. The hazards and critical control points identified were high initial contamination of raw foods, poor environmental sanitation and personal hygiene, feeding of overnight moist foods stored at ambient temperature and insufficient steaming of these foods.

Bacillus cereus↗