DNA probes and the detection of food-borne pathogens using the polymerase chain reaction.
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Bacteria↗
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Emerging food-borne pathogens and their significance in chilled foods.
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Aeromonas↗
Antibiotics and the selection of food-borne pathogens.
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Animals↗
Listeria monocytogenes, a food-borne pathogen.
The gram-positive bacterium Listeria monocytogenes is an ubiquitous, intracellular pathogen which has been implicated within the past decade as the causative organism in several outbreaks of foodborne disease. Listeriosis, with a mortality rate of about 24%, is found mainly among pregnant women, their fetuses, and immunocompromised persons, with symptoms of abortion, neonatal death, septicemia, and meningitis. Epidemiological investigations can make use of strain-typing procedures such as DNA restriction enzyme analysis or electrophoretic enzyme typing. The organism has a multifactorial virulence system, with the thiol-activated hemolysin, listeriolysin O, being identified as playing a crucial role in the organism's ability to multiply within host phagocytic cells and to spread from cell to cell. The organism occurs widely in food, with the highest incidences being found in meat, poultry, and seafood products. Improved methods for detecting and enumerating the organism in foodstuffs are now available, including those based on the use of monoclonal antibodies, DNA probes, or the polymerase chain reaction. As knowledge of the molecular and applied biology of L. monocytogenes increases, progress can be made in the prevention and control of human infection.
Animals↗
Identification of two proline transport systems in Staphylococcus aureus and their possible roles in osmoregulation.
The food-borne pathogen Staphylococcus aureus is distinguished from other food-borne pathogens by its ability to grow at water activity values below 0.90. Previous studies have indicated that proline accumulation mediated by transport represents a primary osmoregulatory strategy utilized by this bacterium (C. B. Anderson and L. D. Witter, Appl. Environ, Microbiol. 43:1501-1503, 1982; I. Koujima, H. Hayashi, K. Tomochika, A. Okabe, and Y. Kanemasa, Appl. Environ. Microbiol. 35:467-470, 1978; K. J. Miller, S. C. Zelt, and J.-H. Bae, Curr. Microbiol. 23:131-137, 1991). In this study, we demonstrate the presence of two proline transport systems within whole cells of S. aureus, a high-affinity transport system (Km, 7 microM) and a low-affinity transport system (Km, 420 microM). Our results indicate that the low-affinity proline transport system is osmotically activated and is the primary system responsible for the accumulation of proline by this pathogen during growth at low water activity.
Betaine↗
Inhibition of food-borne bacterial pathogens by bacteriocins from lactic acid bacteria isolated from meat.
Ten strains of bacteriocin-producing lactic acid bacteria were isolated from retail cuts of meat. These 10 strains along with 11 other bacteriocin-producing lactic acid bacteria were tested for inhibitory activity against psychotrophic pathogens, including four strains of Listeria monocytogenes, two strains of Aeromonas hydrophila, and two strains of Staphylococcus aureus. Inhibition due to acid, hydrogen peroxide, and lytic bacteriophage were excluded. The proteinaceous nature of the inhibitory substance was confirmed by demonstration of its sensitivity to proteolytic enzymes. Eight of the meat isolates had inhibitory activity against all four L. monocytogenes strains. Bacteriocin activity against L. monocytogenes was found in all of the strains obtained from other sources. Activity against A. hydrophila and S. aureus was also common.
Aeromonas↗
Inhibition of PCR by components of food samples, microbial diagnostic assays and DNA-extraction solutions.
We have tested the influence on the polymerase chain reaction (PCR) of a large number of compounds found in food, in media used for selective propagation of food-borne pathogens or in DNA-extraction methods. PCR was found to be sensitive to large volumes of complex food samples containing high amounts of fat and protein, however, an extraction procedure based on treatment with hot NaOH/SDS reduced the effect significantly. Some culture media (Fraser, MLEB, MRB and Rappaport) interfered with the analysis and for most of the media it was possible to assign the inhibitory effect to one or more individual components. Several compounds (detergents, lysozyme, NaOH, alcohols, EDGA, EGTA) used in DNA extraction procedures were found to have some inhibitory effect. The inhibitory effects need to be taken into consideration when designing new tests.
Animals↗
Incidence of toxigenic vibrios in foods available in Taiwan.
A total of 1088 vibrios and related species were isolated from seafood and aquacultured foods available in Taiwan. They were identified as Vibrio alginolyticus, V. cholerae, V. fluvialis I, V. fluvialis II, V. parahaemolyticus, V. mimicus, Aeromonas caviae, A. hydrophila, A. sobria and other species. Incidence of these Vibrio and Aeromonas species in these foods was high. Vibrio parahaemolyticus was frequently found in seawater and in foods of freshwater origin. The Vibrio isolates were examined for enzymatic and toxigenic activities. Most of them showed strong lipase or protease activities. Haemolytic activities of V. cholerae, V. fluvialis I and V. fluvialis II isolates were mostly strong. About 49% showed cytotoxic activity and 5% cytotonic activity in Chinese hamster ovary cell culture assay. Nevertheless, only three non-O1 V. cholerae (2.07%) and two V. parahaemolyticus isolates (1.65%) produced cholera toxin and thermostable direct haemolysin activity, respectively. Various toxigenic vibrios may be important food-borne pathogens in this region because of their high incidence in foods.
Aeromonas↗
Survival of salmonellae during pepperoni manufacture.
Survival of salmonellae in artificially contaminated beef-pork mixtures (approximately 10(4) salmonellae/g) was studied in pepperoni prepared by either a natural flora or lactic starter culture fermentation or in nonfermented sausages. The pepperoni did not become salmonellae free during the usual commercial 15 to 30-day drying period. Salmonella dublin was present in all products, fermented or unfermented, after 42 to 43 days of drying. At a lower level of contamination, 10(3)/g, S. dublin could not be recovered from starter culture-fermented pepperoni after 14 days of drying but persisted in the natural flora-fermented sausage. S. typhimurium (initial count, 10(4)/g) was absent after 42 days of drying when starter culture was used to ferment the pepperoni, but was still present in the natural flora-fermented and unfermented products. S. dublin, host adapted to cattle, or S. choleraesuis, host adapted to swine, had similar survival patterns in beef pork, or beef-pork pepperoni. Heating salmonellae contaminated beef-pork pepperoni (after fermantation but before drying) to an internal temperature of 60 C (trichinae inactivating) eliminated the food-borne pathogen from the sausage product.
Animals↗
Infected aneurysm of the abdominal aorta due to Listeria monocytogenes.
A 79-yr-old man was known for a year with a deteriorating clinical condition, vague abdominal complaints and an elevated erythrocyte sedimentation rate; he was afebrile. Extensive evaluation revealed no cause for his progressive disease. Eventually an infected aneurysm of the abdominal aorta was diagnosed, from which Listeria monocytogenes was cultured. After resection of the aneurysm the patient recovered initially very well. Regrettably, therapy-resistant chylous ascites developed, and the patient died due to surgical complications following a second laparotomy. Infected aortic aneurysms can present as an insidious disease, which may have catastrophic consequences if undiagnosed. A high index of suspicion is required to make a correct diagnosis. L. monocytogenes is an emerging, food-borne pathogen that can cause a wide spectrum of human diseases.
Aged↗
Characterization of a novel putative lantibiotic biosynthesis genomic island in emerging clones of Listeria monocytogenes serotype 4b.
Listeria monocytogenes is a Gram-positive facultative intracellular bacterium that is ubiquitous in nature and the causative agent of listeriosis. The outbreak-derived serotype 4b strain L. monocytogenes strain WS1, sequence type (ST) 558, sublineage (SL) 558, was previously found to have unusual pathogenicity, with ability to cause fetal damage in the first trimester of pregnancy. Search of the WS1 genome for novel and unique genomic features identified a putative lantibiotic island on the chromosome of WS1 and all tested strains of SL558 and two other putative emerging serotype 4b clones, clonal complex 554 (SL554 and SL555) and ST782 (SL782), but absent from all other major clones of L. monocytogenes. The island was deleted from four strains, including two each of ST558 and ST554. The deletions did not impact virulence in a Galleria mellonella model but consistently resulted in reduced hemolytic activity. In addition, we noted strain-dependent impacts on biofilm formation. Additional studies will be necessary to further elucidate the roles of this genomic island in the adaptive physiology and virulence of L. monocytogenes.
Listeria monocytogenes↗
[Bacteriologic quality of the prepared dishes in cafeterias].
In this work, we reports results of bacteriologic analysis of 352 cooked meals (vegetables, meats, desserts). Although, 32% of analysed samples are bacteriologically good, 68% are contamined either by S.P.C but potentially dangerous either by pathogenic bacteria involved in food-born infections.
Algeria↗
[Enterohemorrhagic Escherichia coli O157:H7, an underestimated food pathogen? A literature review].
Escherichia coli O157:H7 has recently been recognised as a human pathogen involved in outbreaks and sporadic cases of food-borne diseases in North America and Great Britain. At present, less is known about the significance of E. coli O157:H7 as a cause of food-borne diseases in the Netherlands. This article will review current literature about this serotype, which is a predominant representative of the enterohaemorrhagic E. coli (EHEC) group. Biochemical features, toxins, pathogenesis, clinical characteristics, epidemiology and methods of isolation will also be discussed.
Animals↗
Comparison of methods for discrimination between strains of Listeria monocytogenes from epidemiological surveys.
Total cellular DNA from 28 strains of Listeria monocytogenes isolated from food implicated in food-borne illness and from patients with listeriosis was digested with the restriction endonucleases HindIII, HaeIII, and EcoRI. Following agarose gel electrophoresis, the fragments were subjected to Southern blot hybridization with a digoxigenin-labeled cDNA probe transcribed from Escherichia coli 16S and 23S rRNA. The patterns of bands from genomic (DNA fingerprints) and rDNA fingerprints (ribotypes) were used for classifying L. monocytogenes strains, and the resulting subtypes were compared with serotyping and multilocus enzyme electrophoresis classification schemes. A total of 15 distinct and identical groups were obtained when genomic DNA was digested with either HindIII or HaeIII. The most discriminating enzyme for ribotyping of strains was EcoRI, which divided the 28 strains of L. monocytogenes into 6 ribotype groups. DNA fingerprinting and ribotyping differentiated L. monocytogenes from other Listeria spp., including L. ivanovii, L. welshimeri, and L. innocua as well as the lactic acid bacteria Lactococcus lactis subsp. lactis and subsp. cremoris. L. monocytogenes strains isolated from four independent food-borne illness incidents were analyzed by all typing methods. Patient and product isolates were not distinguishable by serotyping, ribotyping, or multilocus enzyme electrophoresis. DNA fingerprinting was the only method capable of differentiating these strains, or conversely, of proving relatedness of patient-product pairs of isolates. This method was a relatively simple, sensitive, reproducible, and highly discriminating method for epidemiological tracking of L. monocytogenes implicated in food-borne illness.
Animals↗
Selective plating medium for quantitative recovery of food-borne Listeria monocytogenes.
A new plating medium (lithium chloride-ceftazidime agar [LCA]) was designed to quantitatively recover food-borne Listeria monocytogenes in the form of large colonies while inhibiting most other food-borne microorganisms. This medium included brain heart infusion agar as the nutritive agar base and a combination of selective agents (LiCl, glycine anhydride, and ceftazidime). Comparison of LCA and lithium chloride-phenylethanol-moxalactam agar (LPM) indicated that both were equally effective for the enumeration of the cold-tolerant pathogen in artificially and naturally contaminated foods. However, LCA was more effective than LPM in the recovery of sublethally heat-injured cells. Moreover, Listeria colonies on LCA exhibited a more distinct bluish hue than those on LPM when viewed by the Henry oblique transillumination technique.
Culture Media↗
[Contamination of enteral diet solutions in nosocomial environment].
The gastrointestinal tract is comparatively resistant to food-born germs, but recent studies suggest that nosocomial infections may be triggered by this route. In a study with industrialized diets, prepared with aseptic technique and stored up to 24 hours, aerobic and anaerobic contaminants were searched. Samples were taken after 0.8 and 24 hours, whereas half of these last two analyses were carried out in material left at room temperature, and the other half in refrigerated diets. Initial examination revealed 50% of positive cultures, but part of this was due to non-pathogenic Bacillus germs. After 8 and 24 hours 90% of the samples grew organisms, again with a large proportion of Bacillus, but also with several Gram-negative bacteria, as well as rare Gram-positives. Diarrhea and fever were not registered in patients submitted to enteral nutrition during the study period, nor could any episodes of bacteremia or septic shock be attributed to contaminated feeding material. This lack of clinical consequences of the reported bacterial isolations is not unexpected, and suggests that low concentrations of microorganisms were probably present in the preparations, below a critical level. Nevertheless, attention will be required in the future for better quality control of enteral nutrition mixtures, specially when resistant strains of Gram-negative species are identified, and also in the management of debilitated or immunologically compromised hosts.
Bacteria, Aerobic↗
Enterotoxigenic enteric bacteria in foods and outbreaks of food-borne diseases in Sweden.
All of 86 food routinely examined for potentially pathogenic enteric bacteria were found to harbour one or more coliform species. None of the strains isolated produced heat-labile enterotoxin (LT) or showed invasive properties. The suckling mouse test indicated that one strain of Escherichia coli produced heat-stable enterotoxin (ST). Twelve incidents of suspected food poisoning were also investigated. In two of them the foods examined contained LT-producing strains of E. coli and in two there were LT-producing strains of Klebsiella pneumoniae. The counts of viable enterotoxigenic micro-organisms in these foods were 3000-30,000 E. coli/g and 50,000 to 1 million K. pneumoniae/g. The dominant symptom in all the incidents was watery diarrhoea. These seem to be the first reported cases of foodborne enterotoxigenic enteric bacteria in Europe. Though enterotoxigenic E. coli and related gram-negative enterotoxin-producing species are rare in correctly handled food in Sweden, these micro-organisms should be searched for when outbreaks of food poisoning are investigated.
Enterotoxins↗
Hazard analysis critical control point (HACCP) in public catering services: a modified method, combined to bacteriologic assay.
During 1990 and 1991 the Veterinary and Public Health Services of USL 35 of Ravenna carried out a research programme aimed at the control of food-borne diseases in the sector of public catering services, in collaboration with the Public Health Laboratory (Presidio Multizonale di Prevenzione). The objectives were: obtaining sure information about health hazards in public catering services; checking structural characteristics and equipment of workrooms in restaurants, hotels and refectories; verifying food preparation and preservation methods; promoting health education to increase employees' awareness of hygiene-related problems. The first objective, evaluation of the level of the control of the workrooms exerted on the food contamination hazard by pathogenic or potentially pathogenic organisms, was carried out by allotting specific scores to several characteristics of laboratories or workers' habits, as suggested by the "Hazard Analysis Critical Control Point" (HACCP) method for butcher's shops and fish markets. Five hundred ninety-eight public catering service units have been inspected in restaurants, hotels, school-refectories, factories, hospitals and social houses; 2,097 bacteriological examinations by agar-contact plates and swabs were carried out; 118 preserved-food temperatures were measured, especially in deep-frozen and cooked food; 70 food specimens were tested to search for Salmonella spp. and Staphylococcus aureus and measure Total Aerobic Mesophilic Weight. The presence of Enterobacteriaceae and Escherichia coli was also tested.
Bacteriological Techniques↗