[Show abstract][Hide abstract] ABSTRACT: The MOPC 467 Bio-Enzabead™ enzyme immunoassay (EIA) identified 335 (92.8%) of 361 Salmonella strains tested and gave false positive reactions with 5 (6.5%) of 77 strains of nonsalmonellae. Lack of specificity was particularly prominent with S. typhi but not restricted to any one of the 25 somatic groupings represented in this study. Four of the five false-positive reactions were attributed to Citrobacter freundii. The EIA assay identified 27 (96%) of 28 naturally contaminated foods and produced 16 (25%) false positive reactions which were mainly associated with Citrobacter spp. Attempts at method brevity underlined the importance of M broth post-enrichment and centrifugation for maximum method sensitivity and the inadequacy of EIA for detection of Salmonella in preenrichment and enrichment cultures.
No preview · Article · Aug 2006 · Journal of Food Science
[Show abstract][Hide abstract] ABSTRACT: Conventional isolation methods, including the Health Products and Food Branch (HPFB), Health Canada method used for the isolation and identification of Listeria species and Listeria monocytogenes from foods and environmental samples, can take a week or more to complete and are usually labor-intensive. This has led to the development of various rapid methods which attempt to generate results comparable to standard methods but in a reduced time-frame with less hands-on operation. Our previous work with rapid detection systems indicated that the recommended enrichment protocols failed to grow the Listeria to detectable levels in a reliable and consistent manner. In the present study, a novel enrichment protocol is described and consists of samples being pre-enriched in Palcam broth (incubated at 35 degrees C for 26 h), enriched in UVM 2 (30 degrees C for 26 h) and then plated and analysed by a rapid detection kit, with results being generated after 52 h of incubation. In total, 200 naturally contaminated samples were analysed by both the HPFB standard method and the Palcam method. The results showed that the Palcam method is comparable to the HPFB method. Further analysis involved a rapid detection system, which applies ELISA techniques and automation in an enzyme-linked fluorescent assay (ELFA) system. This system, referred to as the Vitek Immuno Diagnostic Assay System or VIDAS, can identify Listeria to the genus or species (L. monocytogenes) level. In this comparison, an additional 324 naturally contaminated samples were analysed by both the Palcam and ELFA methods. The sensitivity and specificity of the ELFA method were 98.1% and 97.0%, respectively, while the efficiency was 97.5%. False-negative and false-positive rates were 1.9% and 3.0%, respectively. These results show that the ELFA method (when using the Palcam method for pre- and secondary enrichment) was efficient and gave reliable results after 52 h of incubation, and met Health Canada's criteria for approval as a rapid method.
No preview · Article · Apr 2003 · International Journal of Food Microbiology
[Show abstract][Hide abstract] ABSTRACT: The prevalence of microwave ovens in North American homes has increased dramatically within the last decade. Although microwave ovens are primarily used for reheating of foods, they are now more commonly being applied to the cooking of raw foods. Although cooking of raw foods, according to manufacturers' instructions targets an organoleptically acceptable end product, the process does not address the microbiological safety of the cooked food. Seventeen microwave ovens from various commercial suppliers were used to cook naturally contaminated whole raw broilers (< or = 1.8 kg) and roasters (> 1.8 kg) according to manufacturers' instructions. Temperature probes (six per chicken) were used to measure the temperature of chickens immediately after cooking and during the holding period. Of 81 Listeria-positive raw broilers and 93 raw roasters, 1 (1.2%) and 9 (9.7%), respectively, yielded viable Listeria spp. after microwave cooking. Of these, two were undercooked (visual inspection), one was over the maximum weight stipulated by the oven manufacturer and another one was over the maximum weight and undercooked. A significantly greater proportion of contaminated cooked birds was observed with roasters than with broilers, where for one of these contaminated roasters, the temperature at all six measured sites was > or = 87 degrees C. Most of the postcook Listeria-positive birds were associated with 2 of the 17 microwave ovens. Factors such as wattage, cavity size, and the presence or absence of a turntable seemingly did not play a significant role in the survival of Listeria spp. in microwave-cooked chicken. However, the general inability of microwave ovens to uniformly heat chicken carcasses was noted. In order to promote greater safety of microwave-cooked foods, general recommendations for consumers are provided.
No preview · Article · Dec 1998 · Journal of food protection
[Show abstract][Hide abstract] ABSTRACT: A performance assessment of the GENE-TRAK® colorimetric probe assay using pure cultures and naturally contaminated foods and animal feeds underlined the high sensitivity and specificity of this DNA-rRNA diagnostic system. The probe effectively identified 110 (100%) strains of Salmonella spp, and yielded no false-positive reactions in the examination of 61 pure cultures of nonsalmonellae. Of the 53 contaminated raw meats and other high-moisture samples examined in this study, 47 (88.7%) were detected using the GENE-TRAK® analytical protocol. The system also identified 21 (95.5%) of 22 contaminated low-moisture foods and animal feeds. The overall sensitivity and specificity of the GENE-TRAK assay procedure with the naturally contaminated foods examined in this study was 90.7 and 100%, respectively. Ancillary work showed that the choice of selective enrichment conditions played a determinant role in the performance of the probe system. Although the GENE-TRAK protocol relies on varying temporal regimens of selective enrichment in tetrathionate brillant green (TBG35) and selenite cystine (SC35) and postenrichment in gram-negative (GN) broth for high and low moisture food products, our results suggest that, irrespective of product type, probing of GN (6-h) postenrichment cultures inoculated from homologous TBG43 and SC35 (24-h) cultures and subsequent combination of these postenrichment cultures into a single probe assay would enhance the performance of the GENE-TRAK system to a level of unfailing sensitivity and specificity. Attempts at method brevity through direct probing of TBG43, TBG35, SC35, and Rappaport-Vassiliadis (RV43) enrichment cultures proved unsuccessful where probing of short (6-h) enrichment cultures identified ≤15.1% and ≤72.7% of contaminated high- and low-moisture foods, respectively. Direct probing of 24 h enrichment cultures yielded homologous values of ≤17.0% and ≤77.3%. These findings suggest that components in the selective enrichment broths inhibit the probe assay. This negative effect was most prominent with RV43 enrichment cultures where most samples known to contain Salmonella spp. yielded false-negative reactions. Treatment of RV43 cultures, with 0.1 M sodium citrate prior to probe assay was partially effective in neutralizing the inhibitory agent(s).
No preview · Article · Sep 1995 · Journal of food protection
[Show abstract][Hide abstract] ABSTRACT: Refrigerated preenrichment 72 h and selective enrichment cultures arising from 25 g analytical units of dry foods can be used to increase the analytical flexibility and productivity of laboratories for the detection of foodborne Salmonella spp. by AOAC method 994.04. Results of this intralaboratory study using artificially contaminated dry foods validate the extended application of the refrigerated preenrichment approach to dry food composites (375 g). All samples found to be contaminated by AOAC/Bacteriological Analytical Manual methods were identified readily from the homologous, refrigerated preenrichment broth cultures. This extended application of the refrigeration approach was recently adopted First Action by AOAC and was included as a modification to method 994.04. In addition, ancillary work on the diagnostic value of prolonged (48 h) incubation of lysine iron (LI) agar as described in the AOAC Official Method 967.26 led to a recommendation that the 48 h period of incubation be revoked in favor of a 24 h incubation of inoculated LI medium.
No preview · Article · Sep 1995 · Journal of AOAC International
[Show abstract][Hide abstract] ABSTRACT: A collaborative study was conducted to compare the productivity of refrigerated pre-enrichment and enrichment broth cultures with the U.S. Food and Drug Administration culture methods for detection of Salmonella. The refrigerated pre-enrichment and selective enrichment broth culture methods for detection of Salmonella in dry foods have been adopted first action by AOAC INTERNATIONAL.
No preview · Article · Nov 1994 · Journal of AOAC International
[Show abstract][Hide abstract] ABSTRACT: An interlaboratory study was performed in 11 laboratories to validate the use of pre-enrichment and tetrathionate brilliant green (TBG35) and selenite cystine (SC35) enrichment cultures refrigerated 72 h at 2-5 degrees C for greater analytical flexibility in the detection of Salmonella in dry foods. Productivities of refrigerated pre-enrichment and enrichment cultures were compared with that of the AOAC/Bacteriological Analytical Manual (BAM) procedure using 4 food types: whole egg powder, milk chocolate, animal feed, and instantized skim milk powder. Uninoculated and inoculated samples were included in each food group. There was complete agreement between the results obtained by the standard AOAC/BAM procedure and the 2 refrigeration procedures. Of 660 samples tested, the AOAC/BAM procedure identified 393 contaminated samples that were readily detected from the corresponding refrigerated pre-enrichment cultures and from the combined productivity of homologous refrigerated TBG35 and SC35 cultures. Refrigeration (72 h) of pre-enrichment or enrichment cultures for greater analytical flexibility and laboratory productivity in the examination of dry foods is under review for adoption by AOAC International.
No preview · Article · Jul 1993 · Journal of AOAC International
[Show abstract][Hide abstract] ABSTRACT: The sensitivity of the standard cultural method of the International Organization for Standardization (ISO 6579 and ISO 3565 combined) was compared to that of the Health Protection Branch (HPB) procedure for the detection of foodborne Salmonella. Of 195 foods tested, 84 (43.1%) were found to contain salmonellae by one or more cultural conditions. Of these, 75 (89.3%) and 68 (81.0%) were identified by the ISO and HPB methods, respectively. The apparent lack of agreement between both methods likely stemmed from the low indigenous numbers of salmonellae in several food homogenates, and unequal transfer of the target microorganism into homologous ISO and HPB pre-enrichment broths. The sensitivities of the commercially available Muller-Kauffmann tetrathionate broth (MKTBG43, Oxoid CM343), and a closely-related medium prepared with Oxoid CM29 tetrathionate base varied from 86.9 to 89.3%, and were deemed equivalent to that obtained with the ISO formulation of MKTBG43 (89.3%). Comparatively fewer contaminated samples were identified from selenite cystine (SC35) and selenite brilliant green (SBG35) enrichment cultures (82.1-83.3%). The high selectivity and saccharide-independent response of the bismuth sulfite agar medium warrants its consideration as a mandatory plating medium in ISO methodologies for the effective detection of typical and atypical biotypes of foodborne Salmonella spp.
No preview · Article · Jun 1992 · International Journal of Food Microbiology
[Show abstract][Hide abstract] ABSTRACT: The effect of prolonged (48 h) incubation on the productivity of five enrichment-temperature conditions (tetrathionate brilliant green, 35 and 43 degrees C; Muller-Kauffman tetrathionate brilliant green, 43 degrees C; Rappaport-Vassiliadis, 43 degrees C; selenite cystine, 35 degrees C) was compared to homologous results obtained under standard (24 h) conditions of selective enrichment. Of 797 high moisture and 166 low moisture foods tested, 171 (21.5%) and 80 (48.2%), respectively, were found to contain salmonellae by one or more analytical condition. Combined results of the five enrichment conditions after 24 and 48 h of incubation identified 247 (98.4%) and 250 (99.6%) of the 251 contaminated samples identified in this study. Our results are at variance with earlier reports on the greater method sensitivity with extended (greater than or equal to 48 h) periods of selective enrichment. The productivities of individual enrichment conditions after each period of incubation varied markedly where recovery rates with TBG43 and MKTBG43 exceeded that obtained with SC35 and TBG35. Our findings also underline the determinant role of enrichment at an elevated temperature (43 degrees C), and use of multiple enrichment and plating media for the optimal recovery of foodborne Salmonella.
No preview · Article · Jan 1992 · International Journal of Food Microbiology
[Show abstract][Hide abstract] ABSTRACT: The occurrence of Salmonella spp. in red-eared (Pseudemys scripta elegans) turtle eggs imported into Canada from Louisiana in June to September 1988 was examined. Of 28 lots tested, six (21%) lots from three of four exporters harbored salmonellae. Salmonella poona and Salmonella arizonae were frequently encountered in both fertile eggs and packaging moss. Turtles hatched in our laboratory from affected lots of eggs shed Salmonella in tank water for up to 11 months. Widespread use of gentamicin on turtle farms to produce Salmonella-free eggs for export apparently encouraged development of antibiotic resistance in bacterial strains. Of 37 Salmonella strains isolated in this study, 30 (81%) were gentamicin resistant. Such high levels of antibiotic-resistant salmonellae in turtle eggs pose a serious human health risk. Further marketing of turtle eggs and hatchlings should be curtailed until consistent production and distribution of Salmonella-free stocks can be assured.
No preview · Article · Sep 1990 · American Journal of Epidemiology
[Show abstract][Hide abstract] ABSTRACT: Rehydration and equilibration (4 h) of feeds and feed ingredients at water/sample ratios (vol/wt) of 1.4 to 3.2 did not markedly increase recovery of Salmonella spp. in the slurry when analyzed by standard cultural and direct enrichment methods. Of 143 naturally contaminated samples examined, equilibration increased levels of detection from 106 to 109 positive samples by the standard cultural method and from 103 to 112 by direct enrichment. Results suggest that nonhomogeneous distribution of low incident numbers of salmonellae in test samples rather than an equilibration-dependent response provided for the observed heterogeneity in recovery patterns. The novel equilibration approach is of limited application and requires validation on an individual food basis.