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An efficient sampling technique used to detect four foodborne pathogens on pork and beef carcasses in nine Belgian abattoirs.

The method presented in this paper should prove useful in assessing the effectiveness of HACCP plans developed in slaughterhouses. Samples were collected by swabbing well-defined areas of pork and beef carcasses with sterile gauze. Between 160 and 420 half-carcasses were swabbed in each of nine pork or beef slaughterhouses. Swabs from five carcasses were placed in the same sterile Stomacher bag, constituting a single composite sample. Standard or validated analytical methods were used to isolate and characterize four foodborne pathogens. Salmonella spp., Listeria monocytogenes, Campylobacter spp., and verocytotoxin-producing E. coli were detected, respectively, in 27, 2, 2, and 14% of the pork samples and 0, 22, 10, and 5% of the beef samples. Of the 10 samples positive for E. coli O157, only one yielded an isolate confirmed to be enterohemorrhagic. Since Salmonella spp. appear as the main contaminant port (27%) and L. monocytogenes as the main containment of beef (22%), any slaughterhouse sampling plan should include testing for the former in the case of pork carcasses and for the latter in the case of beef carcasses. One should also test regularly for the presence of E. coli O157 and Campylobacter spp. in pork and beef abattoirs. The method presented here is an easy way to assess the contamination rate of carcasses at the end of the slaughtering process.

Abattoirs↗

Injury recovery of foodborne pathogens in high hydrostatic pressure treated milk during storage.

Bacteria are expected to be injured or killed by high hydrostatic pressure (HHP). This depends on pressure levels, species and strain of the microorganism and subsequent storage. Injured bacteria may be repaired which could affect the microbiological quality of foodstuffs with an important safety consideration especially in low acid food products. In this study two Gram-positive (Listeria monocytogenes CA and Staphylococcus aureus 485) and two Gram-negative (Escherichia coli O157:H7 933 and Salmonella enteritidis FDA) relatively pressure resistant strains of foodborne pathogens were pressurized at 350, 450 and 550 MPa in milk (pH 6.65) and stored at 4, 22 and 30 degrees C. The results of shelf life studies indicated two types of injury, I1 and I2, for all the pathogens studied. It is obvious that I2 type injury is a major injury and after its repair (I2 to I1), the cells can form colonies on non-selective but not on selective agar. The formation of colonies on both selective and non-selective agar occurs only after full recovery of injury (I1 to AC). The results presented in this study show that even if injured cells are not detected immediately after HHP treatment, I2 type injury could be potentially present in the food system. Therefore, it is imperative that shelf life studies must be conducted over a period of time for potential repair of I2 type injury either to detectable injury (I1) or to active cells (AC) to ascertain microbiological safety of low acid food products.

Adaptation, Physiological↗

Evolution of foodborne pathogens via temperate bacteriophage-mediated gene transfer.

Temperate bacteriophages have always been central to the evolution of bacteria, although their importance has been consistently underestimated compared to transformation and conjugation. In the last 20 years, as more gene and genome sequences have become available and researchers have more accurately determined bacteriophage populations in the environment, we are gaining a clearer picture of their role in the past and potential role in the future. The transductive and lysogenic capacities of this class of bacteriophages have contributed to the evolution and shaping of emerging foodborne pathogenic bacteria through the dissemination of virulence and antibiotic resistance genes. For example, the genome sequences of Shigella dysenteriae, Escherichia coli O157:H7, and the Stxencoding bacteriophages demonstrate the critical role bacteriophage-mediated gene transfer events played in the evolution of these high-profile human pathogens. In this review, we describe the basic genetic exchange mechanisms mediated by temperate bacteriophages and how these mechanisms have been central to the dissemination of virulence genes, such as toxins and antibiotics from one species to another (the shiga-like toxins, and multiple antibiotic resistance dissemination in Salmonella are used as specific examples). Data demonstrating the role of bacteriophages in the spread of antimicrobial resistance in bacteria, including interspecies transduction, are also presented. That temperate bacteriophages play a role in the on-going evolution of emerging pathogenic bacteria is obvious, but it is also clearly an on-going process with a breadth that must be appreciated as well as studied further if we are to be able to foresee what new challenges will arise to imperil food safety.

Bacterial Infections↗

Evaluation of detection methods for screening meat and poultry products for the presence of foodborne pathogens.

Rapid and molecular technologies such as enzyme-linked immunosorbent assay (ELISA), PCR, and lateral flow immunoprecipitation can reduce the time and labor involved in screening food products for the presence of pathogens. These technologies were compared with conventional culture methodology for the detection of Salmonella, Campylobacter, Listeria, and Escherichia coli O157:H7 inoculated in raw and processed meat and poultry products. Recommended protocols were modified so that the same enrichment broths used in the culture methods were also used in the ELISA, PCR, and lateral flow immunoprecipitation assays. The percent agreement between the rapid technologies and culture methods ranged from 80 to 100% depending on the pathogen detected and the method used. ELISA, PCR, and lateral flow immunoprecipitation all performed well, with no statistical difference, compared with the culture method for the detection of E. coli O157:H7. ELISA performed better for the detection of Salmonella, with sensitivity and specificity rates of 100%. PCR performed better for the detection of Campylobacter jejuni, with 100% agreement to the culture method. PCR was highly sensitive for the detection of all the foodborne pathogens tested except Listeria monocytogenes. Although the lateral flow immunoprecipitation tests were statistically different from the culture methods for Salmonella and Listeria because of false-positive results, the tests did not produce any false negatives, indicating that this method would be suitable for screening meat and poultry products for these pathogens.

Animals↗

Application of the genome sequence to address concerns that Mycobacterium avium subspecies paratuberculosis might be a foodborne pathogen.

Johne's disease, a chronic inflammatory disease caused by infection with Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis), is one of the most prevalent and costly diseases of dairy cattle worldwide. This ruminant pathogen is closely related to the ubiquitous animal and human pathogen Mycobacterium avium subspecies avium (M. avium), confounding the development of specific diagnostic reagents. Exacerbating this problem further is that most existing microbiological, serological, and immunologic assays for the identification of infected animals are inadequate. This is primarily because of the slow-growing nature of the organism, genetic intractability and the previous lack of information on M. paratuberculosis subspecies-specific genes or proteins that may enable the development of specific and sensitive assays. New detection tools are critically needed to definitively answer questions surrounding M. paratuberculosis as a foodborne pathogen as well as aid in determining if it is a contributing factor in Crohn's disease. Thus, the recent characterization of the complete genome sequence of M. paratuberculosis in our laboratories has been a major step forward in meeting this need. We have performed studies that utilize genomic information for the identification of specific DNA sequences and protein antigens in M. paratuberculosis. Based on a preliminary in silico comparison of the M. paratuberculosis genome sequence with that of M. avium, we have now identified at least 35 novel coding sequences that are unique to M. paratuberculosis. These in silico data were then confirmed and expanded by PCR amplification analysis with DNA from several species and isolates of mycobacteria. Finally, these unique sequences have been incorporated into an antigen discovery project that may allow reliable detection of the bacterium in antigen-based diagnostic tests. Application of these new tools in addressing foodborne related issues of M. paratuberculosis is discussed.

Animals↗

Inhibitory activity of honey against foodborne pathogens as influenced by the presence of hydrogen peroxide and level of antioxidant power.

Antimicrobial activity of honey has been attributed to hydrogen peroxide, which is produced by naturally occurring glucose oxidase, and phenolic compounds, although lethality of and inhibition by these and other components against microorganisms vary greatly, depending on the floral source of nectar. This study was undertaken to compare honeys from six floral sources for their inhibitory activity against Escherichia coli O157:H7, Salmonella typhimurium, Shigella sonnei, Listeria monocytogenes, Staphylococcus aureus, and Bacillus cereus. A disc assay revealed that development of zones of inhibition of growth depends on the type and concentration of honey, as well as the test pathogen. Growth of B. cereus was least affected. The inhibition of growth of S. sonnei, L. monocytogenes, and S. aureus in 25% solutions of honeys was reduced by treating solutions with catalase, indicating that hydrogen peroxide contributes to antimicrobial activity. Darker colored honeys were generally more inhibitory than light colored honeys. Darker honeys also contained higher antioxidant power. Since antimicrobial activity of the darker colored test honeys was not eliminated by catalase treatment, non-peroxide components such as antioxidants may contribute to controlling the growth of some foodborne pathogens. The antibacterial properties of honeys containing hydrogen peroxide and characterized by a range of antioxidant power need to be validated using model food systems.

Anti-Bacterial Agents↗

Stimulation of starter culture for further reduction of foodborne pathogens during salami fermentation.

This study was conducted to determine if stimulated meat starter culture (MSC; Pediococcus acidilactici) would further control Escherichia coli O157:H7, Listeria monocytogenes, and Staphylococcus aureus during salami fermentation. Manganese ion (0.005% of MnSO4) was used as a stimulator for the growth and acid production of MSC. After 24-h salami fermentation, nonstimulated MSC and stimulated MSC reduced E. coli O157:H7 levels by 1.3 and 2.3 log10 units, respectively. Nonstimulated MSC reduced L. monocytogenes levels by 1.2 log10 units, whereas the stimulated MSC achieved a 2.2-log10 reduction after 24-h fermentation. In the case of S. aureus, nonstimulated MSC and stimulated MSC reduced S. aureus levels by 1.3 and 2.3 log10 units after 24-h fermentation, respectively. Stimulated MSC by MnSO4 reduced those foodborne pathogens more effectively compared with nonstimulated MSC (P < 0.05).

Animals↗

Antibacterial effect of water-soluble arrowroot (Puerariae radix) tea extracts on foodborne pathogens in ground beef and mushroom soup.

Antimicrobial activity of water-soluble arrowroot tea extract was evaluated against Escherichia coli O157:H7, Salmonella enterica Serotype Enteritidis, Listeria monocytogenes, and Staphylococcus aureus in ground beef and mushroom soup. The concentrations of arrowroot tea used were 0, 3, and 6% (wt/wt) for ground beef and 0, 1, 5, and 10% (wt/vol) for mushroom soup. Samples without tea extract were considered controls. Each sample was stored for 0, 1, 3, 5, and 7 days at 7 degrees C for ground beef and for 0, 1, 3, and 5 days at 35 degrees C for mushroom soup. On each sampling time, proper dilutions were spread plated on each pathogen-specific agar. Viable cell counts of each pathogen were performed after incubation at 35 degrees C for 24 to 48 h. For ground beef, Salmonella Enteritidis and L. monocytogenes were slightly suppressed by approximately 1.5 log, compared with the control, on day 7 at 3 and 6% arrowroot tea treatment. For mushroom soup, all test pathogens were suppressed by 6.5, 4.7, 3.4, and 4.3 log at 5% and 6.0, 4.7, 5.0, and 4.3 log at 10% against E. coli O157:H7, Salmonella Enteritidis, L. monocytogenes, and S. aureus, respectively, compared with the control on day 5. Mushroom soup with 1% arrowroot tea also showed 2.3- and 2.7-log growth suppression of Salmonella Enteritidis and S. aureus, respectively, compared with the control on day 5. This study showed that the use of arrowroot tea would effectively inhibit the microbial growth of both gram-negative and gram-positive foodborne pathogens in various foods, especially liquid foods.

Agaricales↗

Impact of molecular biology on the detection of foodborne pathogens.

Molecular biological methods that use antibodies and nucleic acids to detect specific foodborne bacterial pathogens were scarcely known a decade and a half ago. Few scientists could have predicted that these tools of basic research would come to dominate the field of food diagnostics. Today, a large number of cleverly designed assay formats using these technologies are available commercially for the detection in foods of practically all major established pathogens and toxins, as well as of many emerging pathogens. These tests range from very simple antibody-bound latex agglutination assays to very sophisticated DNA amplification methods. Although molecular biological assays are more specific, sensitive, and faster than conventional (often cultural) microbiological methods, the complexities of food matrices continue to offer unique challenges that may preclude the direct application of these molecular biological methods. Consequently, a short cultural enrichment period is still required for food samples prior to analysis with these assays. The greater detection sensitivity of molecular biological methods may also affect existing microbiological specifications for foods; this undoubtedly will have repercussions on the regulatory agencies, food manufacturers, and also consumers.

Antibodies, Bacterial↗

Animal health and foodborne pathogens: enterohaemorrhagic O157:H7 strains and other pathogenic Escherichia coli virotypes (EPEC, ETEC, EIEC, EHEC).

The majority of interactions between microorganisms and animals are based on convenient relations for both of them. Symbiotic microorganisms, like intestinal microbiota, produce important vitamins for animals and protects them from putative pathogens. In general, for monogastric animals, the main contribution of intestinal microorganisms is to supply with growth factors the animal diet, and in some cases they are responsible for providing essential vitamins (e.g. vitamin K). Some particular and relatively few microbes like viruses, bacteria, fungi, protozoa and algae are responsible for animal illness. Because microorganisms are easily dispersed, display physiological diversity, and tolerate extreme conditions, they are ubiquitous and may contaminate and grow in many products, including food and raw materials. Foodborne diseases are caused by consumption of contaminated food or beverages. Many different disease-causing pathogens can contaminate food, so there are many different foodborne infections. In addition, poisonous chemicals and biological toxins can cause disease if they are present in food. To know how a particular disease is spreading is an important matter to take appropriate steps to stop it. For example Escherichia coli O157:H7 infections can spread through contaminated food (meat, vegetables, cheese, etc.), contaminated drinking water or juices, contaminated swimming water and from person to person. Among foodborne pathogens, the most frequently detected are bacteria, but also parasitic protozoa and worms, viruses, natural toxins and other pathogenic agents like prions are important agents for foodborne diseases. Particular pathogenic types of E. coli, classified by their specific pathogenic mechanisms (toxins, adhesins, invasiveness, etc.) are actually known as E. coli virotypes. Enterohaemorrhagic E. coli (EHEC), which constitute the main part of this review, were also named verotoxigenic E. coli (VTEC) or Shiga toxigenic E. coli (STEC). EHEC strains cause haemorrhagic colitis (HC), haemolytic uremic syndrome (HUS) and thrombotic thrombocytopaenic purpura (TP) in humans. They synthetize shigatoxins (verotoxins) which are potent cytotoxic substances, adherence factors and enterohaemolysin. EHEC are responsible for many outbreaks of bloody diarrhoea caused by contaminated foods: beef, milk, fruits, juice, water, etc. The most important serogroups among EHEC are O26, O111 and O157, being O157:H7 the most relevant serotype in foodborne outbreaks. The normal intestinal microflora of cattle was found to be the most relevant reservoir of EHEC strains.

Animals↗

Identification of foodborne pathogens by nucleic acid hybridization.

Nucleic acid hybridization methods have been developed and used to identify microorganisms in foods. Tests performed on mixed cultures save the time required to establish pure cultures. Enterotoxigenic or invasive strains of foodborne bacterial pathogens are detected with probes that identify genes responsible for virulence. Hybridization tests signal the presence or absence of a particular strain or an entire genus and are especially well suited for screening foods for specific pathogens. With the colony hybridization assay format, foodborne bacteria harboring a specific gene can be enumerated. However, hybridization tests require the presence of 10(5) to 10(6) cells to yield a positive result, thereby limiting sensitivity and necessitating a time-consuming growth step. In vitro DNA amplification techniques increase the amount of DNA segments 10(5)-10(6)-fold in 2 to 3 h, thus enhancing test sensitivity.

Bacteria↗

Shigella as a foodborne pathogen and current methods for detection in food.

Shigella, the causative agent of shigellosis or "bacillary dysentery," has been increasingly involved in foodborne outbreaks. According to the Centers for Disease Control and Prevention's Emerging Infections Program, Foodborne Diseases Active Surveillance Network (FoodNet), Shigella was the third most reported foodborne bacterial pathogen in 2002. Foods are most commonly contaminated with Shigella by an infected food handler who practices poor personal hygiene. Shigella is acid resistant, salt tolerant, and can survive at infective levels in many types of foods such as fruits and vegetables, low pH foods, prepared foods, and foods held in modified atmosphere or vacuum packaging. Survival is often increased when food is held at refrigerated temperatures. Detection methods for Shigella include conventional culture methods, immunological methods, and molecular microbiological methods. Conventional culture of Shigella in foods is often problematic due to the lack of appropriate selective media. Immunological methods for Shigella have been researched, yet there is only one commercially available test kit. Molecular microbiological methods such as PCR, oligonucleotide microarrays, and rep-PCR have also been developed for the detection and identification of Shigella. This manuscript reviews the general characteristics, prevalence, growth and survival, and methods for detection of Shigella in food.

Dysentery, Bacillary↗

Emerging foodborne pathogens: keeping your patients and your families safe.

Changes in food production and societal pressures have led to a continuing increase in the incidence of foodborne illness. Many pathogens are associated with specific foods, e.g., E. coli O157:H7 with hamburgers or Salmonella with eggs. The U.S. Food and Drug Administration has recently approved irradiation for sterilization of meat, but public acceptance of irradiated food is low. Because contaminated foods are seldom detected before they reach store shelves, care in food preparation by professional and home cooks is crucial.

Animals↗

Detection of foodborne pathogens using DNA probes and a dipstick format.

The detection of foodborne microorganisms has traditionally been done using microbiologically based methods. Such "gold standard" methods are generally reliable but have the disadvantages of being labor intensive, subjective, and time consuming. Over the last several years, the development of DNA probe-based methods has simplified the methods used to detect organisms such as Salmonella, Listeria, and E. coli by targeting the unique DNA or RNA sequences of these organisms using DNA probes and nonradioactive detection.

DNA Probes↗

Campylobacter jejuni--an emerging foodborne pathogen.

Campylobacter jejuni is the most commonly reported bacterial cause of foodborne infection in the United States. Adding to the human and economic costs are chronic sequelae associated with C. jejuni infection--Guillian-Barré syndrome and reactive arthritis. In addition, an increasing proportion of human infections caused by C. jejuni are resistant to antimicrobial therapy. Mishandling of raw poultry and consumption of undercooked poultry are the major risk factors for human campylobacteriosis. Efforts to prevent human illness are needed throughout each link in the food chain.

Animals↗

Identification, subtyping and virulence determination of Listeria monocytogenes, an important foodborne pathogen.

Listeria monocytogenes is an opportunistic intracellular pathogen that has become an important cause of human foodborne infections worldwide. Given its close relationship to other Listeria species and its tendency to produce non-specific clinical symptoms, the availability of rapid, sensitive and specific diagnostic tests for the differentiation of L. monocytogenes from other Listeria species is helpful for selecting appropriate treatment regimens. In addition, with L. monocytogenes comprising a diversity of strains of varying pathogenicity, the ability to precisely track the strains involved in listeriosis outbreaks and speedily determine their pathogenic potential is critical for the control and prevention of further occurrences of this deadly disease. Extensive research in recent decades has revealed significant insights regarding the molecular mechanisms of L. monocytogenes infection. This in turn has facilitated the development of laboratory procedures for enhanced detection and identification of L. monocytogenes, and has also contributed to the implementation of improved control and prevention strategies against listeriosis. The purpose of this review is to summarize recent progress in the species-specific identification, subtyping and virulence determination of L. monocytogenes strains, and to discuss future research needs pertaining to these important areas of listeriosis.

Animals↗