Occurrence of Campylobacter in retail foods in Ireland

A surveillance study was carried out to determine the prevalence of Campylobacter in a range of retail foods purchased in three Irish cities over a 20-month period between March 2001 and October 2002. In total 2391 food samples were analysed during this period. Campylobacter was isolated from 444 raw chicken (49.9%), 33 turkey (37.5%) and 11 duck samples (45.8%). Lower isolation rates of 7/221 (3.2%), 10/197 (5.1%) and 31/262 (11.8%) were observed for raw beef, pork and lamb, respectively. One sample of pork paté from 120 samples analysed (0.8%) was Campylobacter-positive. A total of three shellfish samples (oysters) from 129 raw specimens examined (2.3%) were found to contain Campylobacter. Low prevalences of the organism (0.9%) were also isolated from fresh mushrooms. Of 62 raw bulk tank milk samples analysed, Campylobacter was recovered in a single sample (1.6%). Campylobacter was not detected in any of the comminuted pork puddings, prepared vegetables and salads, retail sandwiches or cheeses made from unpasteurised milk. In total, 543 Campylobacter were isolated from all of the food samples analysed, of which 453 (83.4%) were confirmed as Campylobacter jejuni and the remaining 90 (16.6%) as Campylobacter coli.     

Campylobacter in ready-to-eat foods: the result of a 15-month survey

Ready-to-eat foods at retail and catering establishments were randomly sampled at the point of sale for a period of 15 months. During this period, 4,469 food samples were examined for the presence of Campylobacter with the use of plate-based methodology. A range of different ready-to-eat food types, all of which have the potential to be contaminated with the organism, were examined. No viable Campylobacter cells were detected in any of the foods during the survey. There is no evidence that the foods examined in this study represent a risk to the consumer. Although these foods may be a vehicle for infection if cross-contamination occurs, other sources need to be investigated as causes of enteric infection caused by Campylobacter.     

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.     

Development and application of a heterologous internal standard for polymerase-chain-reaction-based detection of Campylobacter jejuni and Campylobacter coli in foods

 A heterologous internal standard, termed "mimic", was developed for the polymerase chain reaction (PCR)-based detection of Campylobacter jejuni and Campylobacter coli in food. Mimic was designed to contain a heterologous DNA fragment of plasmid pUC18, flanked by a primer binding site, identical to the bacterial target DNA. Application of mimic in the PCR permitted its co-amplification together with the bacterial DNA with similar efficiency. As the length of the amplified products differed, they were easily detectable by agarose gel electrophoresis. The presence or absence of the mimic PCR product was indicative of the efficacy of the PCR. The use of approximately 60 mimic molecules per reaction was optimal for determining the reliability of the diagnostic PCR assays without decreasing the detection limit. This system for the detection of the two species of Campylobacter was successfully applied in routine food surveillance. Received: 4 January 1999     

Prevalence and numbers of coliphages and Campylobacter jejuni bacteriophages in New Zealand foods

Vegetable samples were tested for the presence of coliphages. None of the 55 samples contained these phages at concentrations greater than 10 g(-1) (the limit of detection). Spiking and recovery experiments indicated that the method was efficient at detecting coliphage T4 added to the food, and so it was concluded that phage titres were not being falsely underestimated. In addition 51 samples of chicken skin from retail portions were tested for the presence and numbers of coliphages and for presence only of Campylobacter jejuni phages. Coliphages were isolated from 46 samples (90.2% positive), at up to 2570 PFU 10 g sample(-1) but no C. jejuni phages were isolated. Several other methods were used to isolate C. jejuni phages from retail chicken but none was successful. However, when pooled whole chicken rinses from 39 flocks were tested for the presence of C. jejuni phages, 11 (28.2%) of the flocks were positive. It is possible that phages present on birds at the start of processing were either inactivated or simply diluted out during spin chilling. These data add to the body of information indicating that phages can readily be isolated from certain foods and indicate that consumers are exposed to them on a regular basis.     

Escherichia coli O157:H7 and its significance in foods

Escherichia coli O157:H7 was conclusively identified as a pathogen in 1982 following its association with two food-related outbreaks of an unusual gastrointestinal illness. The organism is now recognized as an important cause of foodborne disease, with outbreaks reported in the U.S.A., Canada, and the United Kingdom. Illness is generally quite severe, and can include three different syndromes, i.e., hemorrhagic colitis, hemolytic uremic syndrome, and thrombotic thrombocytopenic purpura. Most outbreaks have been associated with eating undercooked ground beef or, less frequently, drinking raw milk. Surveys of retail raw meats and poultry revealed E. coli O157:H7 in 1.5 to 3.5% of ground beef, pork, poultry, and lamb. Dairy cattle, especially young animals, have been identified as a reservoir. The organism is typical of most E. coli, but does possess distinguishing characteristics. For example, E. coli O157:H7 does not ferment sorbitol within 24 h, does not possess beta-glucuronidase activity, and does not grow well or at all at 44-45.5 degrees C. The organism has no unusual heat resistance; heating ground beef sufficiently to kill typical strains of salmonellae will also kill E. coli O157:H7. The mechanism of pathogenicity has not been fully elucidated, but clinical isolates produce one or more verotoxins which are believed to be important virulence factors. Little is known about the significance of pre-formed verotoxins in foods. The use of proper hygienic practices in handling foods of animal origin and proper heating of such foods before consumption are important control measures for the prevention of E. coli O157:H7 infections.     

食品工业用菌的病原性研究

为研究食品工业用真菌的致病性 ,将不同浓度的米曲霉和黑曲霉孢子经小鼠尾静脉注射 ,观察 14d内动物出现的中毒症状、死亡和体重变化 ,实验终结时测定脑、肝、肾、脾中的生存菌数 ,同时作病理组织学检查。结果表明 ,2个菌种对小鼠造成的主要损害为肾脏化脓性病变 ,1× 10 5以上剂量组小鼠 2个以上脏器同时检出活菌。米曲霉 1× 10 5以上剂量组动物出现了中毒症状和死亡 ,黑曲霉实验组动物未见中毒和死亡。致病性随染孢子量的增加而加重 ,呈正剂量 -反应关系。本研究为完善我国食品工业用菌安全性评价体系提供了依据     

Prevalence of thermophilic Campylobacter spp. in ready-to-eat foods and raw poultry in Northern Ireland

Although there have been numerous studies investigating the prevalence of campylobacters in animals and raw meats, there are limited data on the persistence of these organisms in ready-to-eat (RTE) foodstuffs. Although poultry is now well established as a major reservoir of thermophilic campylobacters, it is widely assumed that hazard analysis critical control point (HACCP) controls in commercial and industrial settings are effective in eliminating this hazard through thorough cooking of RTE products. Therefore, it was the primary aim of this study to investigate the effectiveness of HACCP controls in eliminating campylobacters in such cooked RTE foods by attempting to isolate viable organisms from product. Concurrently, the results of this study demonstrate that local poultry is highly contaminated with campylobacters. Commercially available RTE foodstuffs (n = 2,030) consisting of 1,061 poultry-related cooked products and 969 other products were analyzed and were not found to contain thermophilic Campylobacter spp. In addition, 107 raw chickens (63 fresh birds and 44 frozen birds) were sampled, and 94% of the fresh birds and 77% of the frozen birds examined were demonstrated to be contaminated with campylobacters, with Campylobacter jejuni, Campylobacter coli, and Campylobacter lari accounting for 69, 30, and 1% of the contaminating organisms, respectively. In general, commercially available RTE foodstuffs, including cooked poultry, are not commonly contaminated with campylobacters and thus do not appear to represent a significant cause of clinical infection of Campylobacter spp. in Northern Ireland. However, raw poultry produce, including fresh and frozen chicken, frequently tested positive for campylobacters. Implementation of HACCP systems by food processors will help to minimize and/or eliminate the risk posed by this organism to the consumer.     

Helicobacter pylori: characteristics, pathogenicity, detection methods and mode of transmission implicating foods and water

Helicobacter pylori is an organism involved in the pathogenesis of human active chronic gastritis, peptic and duodenal ulcer diseases and gastric cancer. This review article covers this emerging human pathogen in terms of its phenotypic and genotypic characteristics, methods for culturing, its role in gastric pathogenicity, evidence involving its mode of transmission, difficulty in its isolation and detection methodology. In terms of transmission, both foodborne and waterborne pathways have been speculated as the mode of transmission for H. pylori as the patterns of the infection are consistent with those from fecal-oral and oral-oral transmission. Therefore, it is important to also evaluate methods for the detection of H. pylori from specifically food products and water. The detection of this pathogen has proved difficult since changes in cell morphology, metabolism and growth patterns occur when H. pylori is exposed to different environmental stimuli. The development of a viable but non-culturable coccoid (VNC) form is observed. These VNC forms do not undergo cellular division and cannot be cultured by traditional methods, increasing the difficulty in their detection. Since both viability and virulence in the VNC form of H. pylori are retained, the examination of food products and water for these forms is critical. Current methods include filtration, immuno-separation (IMS), polymerase chain reaction (PCR), probe hybridization, immuno-staining, autoradiography and ATP bioluminescence.     

The identification and significance of 2-phenylethylamine in foods.

Techniques are described for the extraction and analysis of amines from foods, employing gas chromatography and mass spectrometry. The presence of 2-phenylethylamine is demonstrated in samples of chocolate, cheese and certain red wines. Subsequent clinical trials have shown that this amine precipitates migraine attack in dietary migraine sufferers.     

The chemistry and biological significance of saponins in foods and feedingstuffs

Saponins occur widely in plant species and exhibit a range of biological properties, both beneficial and deleterious. This review, which covers the literature to mid 1986, is concerned with their occurrence in plants and their effects when consumed by animals and man. After a short discussion on the nature, occurrence, and biosynthesis of saponins, during which the distinction between steroidal and triterpenoid saponins is made, the structures of saponins which have been identified in a variety of plants used as human foods, animal feedingstuffs, herbs, and flavorings are described. Many of these compounds have been characterized only during the last 2 decades, and modern techniques of isolation, purification, and structural elucidation are discussed. Particular consideration is given to mild chemical and enzymatic methods of hydrolysis and to recent developments in the application of NMR and soft ionization MS techniques to structural elucidation. Methods currently used for the quantitative analysis of saponins, sapogenols, and glycoalkaloids are critically considered; advances in the use of newer methods being emphasized. The levels of saponins in a variety of foods and food plants are discussed in the context of the methods used and factors affecting these levels, including genetic origin, agronomic, and processing variables, are indicated. Critical consideration is given to the biological effects of saponins in food which are very varied and dependent upon both the amount and chemical structure of the individual compounds. The properties considered include membranolytic effects, toxic and fungitoxic effects, adverse effects on animal growth and performance, and the important hypocholesterolemic effect. A final section deals briefly with the pharmacological effects of saponins from ginseng, since use of this plant is increasing in certain sections of western society as well as being traditional in the Orient.     

Occurrence of Campylobacter spp. in raw and ready-to-eat foods and in a Canadian food service operation

The occurrence of Campylobacter spp. in a variety of foods from Ottawa, Ontario, Canada, and raw milk samples from across Canada was determined over a 2-year period. The samples consisted of 55 raw foods (chicken, pork, and beef), 126 raw milk samples from raw milk cheese manufacturers, and 135 ready-to-eat foods (meat products, salads, and raw milk cheeses). Campylobacter jejuni was detected in 4 of the 316 samples analyzed: 1 raw beef liver sample and 3 raw chicken samples. An isolation rate of 9.7% was observed among the raw chicken samples tested. This study also investigated the role of cross-contamination in disseminating Campylobacter from raw poultry within a food service operation specializing in poultry dishes. Accordingly, kitchen surfaces within a restaurant in Ottawa, Ontario, were sampled between March and August 2001. Tests of the sampling method indicated that as few as 100 Campylobacter cells could be detected if sampling was done within 45 min of inoculation; however, Campylobacter spp. were not detected in 125 swabs of surfaces within the kitchens of this food service operation. Despite the reported high prevalence of Campylobacter spp. in raw poultry, this organism was not detected on surfaces within a kitchen of a restaurant specializing in poultry dishes.     

Endogenous formaldehyde level of foods and its biological significance

 Formaldehyde was measured in different food samples based on its reaction with hydralazine to form s-triazolo-(3,4-α)-phthalazine(Tri-P). Formaldehyde reacts with hydralazine under mild acidic conditions to form Tri-P. The reaction products were separated by reversed-phase, high-performance liquid chromatography. The method of sample preparation and chromatography are reviewed and the analytical method validated. Using the discussed methods we measured formaldehyde levels in 23 animals and 22 vegetable and fruit samples. Formaldehyde levels were one and a half times higher in vegetable samples compared to meat samples. Received: 23 December 1996 / Revised version: 17 March 1997     

A robotic DNA purification protocol and real-time PCR for the detection of Campylobacter jejuni in foods

Primers designed to amplify a Campylobacter jejuni cadF gene sequence were used in an SYBR Green I real-time PCR assay as an alternative to conventional bacteriological methods for the rapid detection of C. jejuni in foods. Twenty-five grams of chicken skin (breast and thigh) was contaminated by adding approximately 1, 10, or 50 CFU of C. jejuni ATCC 35560. Twenty-five grams of pork and 25-ml aliquots of milk were also inoculated with 1 and 10 CFU of the pathogen. The samples were incubated in Bolton broth for different periods at 37 and 42 degrees C under microaerophilic conditions. Using a commercial robotic DNA purification system, DNA was extracted and purified from 1-ml aliquots of the enrichment cultures before and after centrifugation of the 250-ml enrichment broth at 15,900 x g for 10 min at 4 degrees C. The DNA was used as the template in a real-time PCR assay. C. jejuni was detected after 12 h of enrichment from samples inoculated with about 50 CFU/25-g sample. After centrifugation, an enrichment step of 8 h was sufficient to allow detection of pathogen in samples inoculated with 10 CFU/25 g. However, 24 h of enrichment was necessary to detect pathogen in samples inoculated with approximately 1 CFU/25 g. The real-time PCR protocol developed in this study significantly reduced the detection time of C. jejuni in foods.     

From polyphenol to o-quinone: Occurrence,significance, and intervention strategies in foods and health implications

Polyphenol oxidation is a chemical process impairing food freshness and other desirable qualities, which has become a serious problem in fruit and vegetable processing industry. It is crucial to understand the mechanisms involved in these detrimental alterations. o-Quinones are primarily generated by polyphenols with di/tri-phenolic groups through enzymatic oxidation and/or auto-oxidation. They are highly reactive species, which not only readily suffer the attack by nucleophiles but also powerfully oxidize other molecules presenting lower redox potentials via electron transfer reactions. These reactions and subsequent complicated reactions are capable of initiating quality losses in foods, such as browning, aroma loss, and nutritional decline. To attenuate these adverse influences, a variety of technologies have emerged to restrain polyphenol oxidation via governing different factors, especially polyphenol oxidases and oxygen. Despite tremendous efforts devoted, to date, the loss of food quality caused by quinones has remained a great challenge in the food processing industry. Furthermore, o-quinones are responsible for the chemopreventive effects and/or toxicity of the parent catechols on human health, the mechanisms by which are quite complex. Herein, this review focuses on the generation and reactivity of o-quinones, attempting to clarify mechanisms involved in the quality deterioration of foods and health implications for humans. Potential innovative inhibitors and technologies are also presented to intervene in o-quinone formation and subsequent reactions. In future, the feasibility of these inhibitory strategies should be evaluated, and further exploration on biological targets of o-quinones is of great necessity.     

The public health significance of Aeromonas spp. in foods

There is now evidence that some strains of Aeromonas species are enteropathogens. Such strains possess virulence properties, such as the ability to produce enterotoxins, cytotoxins, haemolysins and/or the ability to invade epithelial cells. Strains with these properties are common contaminants of drinking water and a wide range of foods. Contact or consumption of contaminated water, especially in summer, is a major risk factor in Aeromonas-associated gastroenteritis. Aeromonas-contaminated foods may also be vehicles of infection. Given the properties of strains that have been described in foods it has been suggested that food-borne illness could result not only from colonization and in vivo expression of virulence factors, but possibly also by intoxication following ingestion of foods that have been stored for a period of time, even under refrigeration. This paper reviews what is known about Aeromonas spp. in foods, their expression of virulence determinants, particularly at refrigeration temperatures, and the questions remaining to be answered to evaluate the risk they pose, so that an appropriate public health response can be determined.     

Detection of thermophilic Campylobacter spp. in blood-free enriched samples of inoculated foods by the polymerase chain reaction

The detection of thermophilic Campylobacter spp., as represented by Campylobacter jejuni, by the polymerase chain reaction (PCR) was investigated and compared with the selective agar isolation (SAI) method. Stationary-phase cultures of C. jejuni were inoculated into either blood-free enrichment broth (BFEB) or BFEB that contained 10% broccoli, crabmeat, mushroom, raw milk, and raw oyster rinses. Following a 48-h enrichment period, aliquots of food test portions were removed for simultaneous analysis by PCR and SAI. It was determined that the presence of charcoal and iron in the enrichment broth interfered with the PCR assay. Therefore, three DNA extraction techniques were developed and evaluated using a 16S rRNA primer pair in the PCR assay. The 50% end point (DL50) values (determined upon six initial C. jejuni spiking levels) were used to assess the frequency of isolation utilizing PCR versus SAI for the detection of this organism in the enrichment matrices. There were virtually no differences in detection of C. jejuni among enriched samples analyzed by PCR and SAI. Mean DL50 values (n = 3) for plain BFEB, broccoli, crabmeat, mushroom, raw milk, and raw oyster were, respectively, 0.02 (PCR) versus 0.01 (SAI), 0.01 versus 0.06, 0.07 versus 0.04, 0.03 versus 0.08, 0.01 versus 0.01, and 0.01 versus 0.01 CFU/5 g food. Significant variability in the detection limit of C. jejuni by PCR in the food enrichments was observed among DNA extraction techniques. Using 48-h enrichment cultures followed by PCR analysis could save 1 day of the time required for the presumptive identification of C. jejuni in suspected foods.     

Nanotech: propensity in foods and bioactives

Nanotechnology is seeing higher propensity in various industries, including food and bioactives. New nanomaterials are constantly being developed from both natural biodegradable polymers of plant and animal origins such as polysaccharides and derivatives, peptides and proteins, lipids and fats, and biocompatible synthetic biopolyester polymers such as polylactic acid (PLA), polyhydroxyalkonoates (PHA), and polycaprolactone (PCL). Applications in food industries include molecular synthesis of new functional food compounds, innovative food packaging, food safety, and security monitoring. The relevance of bioactives includes targeted delivery systems with improved bioavailability using nanostructure vehicles such as association colloids, lipid based nanoencapsulator, nanoemulsions, biopolymeric nanoparticles, nanolaminates, and nanofibers. The extensive use of nanotechnology has led to the need for parallel safety assessment and regulations to protect public health and adverse effects to the environment. This review covers the use of biopolymers in the production of nanomaterials and the propensity of nanotechnology in food and bioactives. The exposure routes of nanoparticles, safety challenges, and measures undertaken to ensure optimal benefits that outweigh detriments are also discussed.     

Occurrence of toluene in Canadian total diet foods and its significance to overall human exposure

Levels of most VOCs in foods are usually low because of their volatility, and human exposure to VOCs is expected to be mainly via inhalation of ambient and indoor air. However, dietary exposures to VOCs can be significant to overall exposures if elevated concentrations of VOCs are present in foods consumed in high amounts and/or on a regular basis, and this was demonstrated in this study with the occurrence data of toluene from the recent 2014 Canadian Total Diet Study (TDS). Concentrations of toluene in the composite samples of most food types from the 2014 TDS are low and similar to the results from the previous 2007 TDS with some exceptions, such as beef steak (670 ng/g (2014 TDS) vs. 14 ng/g (2007 TDS)), poultry, chicken and turkey (307 ng/g (2014 TDS) vs. 8.8 ng/g (2007 TDS)). Toluene concentrations in most of the grain-based and fast food composite samples from the 2014 TDS are considerably higher than those from the 2007 TDS, with the highest level of 4655 ng/g found in the composite sample of crackers from the 2014 TDS (compared to 18 ng/g from 2007 TDS). Dietary exposure estimates for toluene based on the occurrence results from the 2014 TDS show that for most of the age groups, grain-based foods are the primary source, accounting for an average of 77.5% of the overall toluene intake from the diet. The highest dietary exposures to toluene were observed for the adult age groups, with estimated average exposures ranging from 177.4 to 184.5 µg/d. Dietary exposure estimates to toluene are well below oral doses associated with toxicological effects and also below the maximum estimated intake (819 µg/d) from air inhalation for adult group (20 – 70 years) based on the results from CEPA (Canadian Environmental Protection Act) assessment in 1992.