Relative Effectiveness of Selective Plating Agars for Recovery of Salmonella Species from Selected High-Moisture Foods

1995 ◽  
Vol 78 (3) ◽  
pp. 679-690 ◽  
Author(s):  
Patricia S Sherrod ◽  
Rene Miguel Amaguana ◽  
Wallace H Andrews ◽  
Geraldine A June ◽  
Thomas S Hammack

Abstract The relative effectiveness of 6 selective plating media were compared for effectiveness in recovery of Salmonella spp. from selected high-moisture foods. Three new plating agars (EF-18, Rambach, and xylose lysine Tergitol-4) and 3 selective plating agars (bismuth sulfite, Hektoen enteric, and xylose lysine desoxycholate) recommended by AOAC INTERNATIONAL and the Bacteriological Analytical Manual (BAM) were compared. The agars were streaked from cultures selectively enriched in selenite cystine broth, tetrathionate broth, and Rappaport–Vassiliadis medium. The high-moisture foods studied were naturally contaminated pork sausage, chicken parts, turkey parts, and frog legs and artificially contaminated shrimp, oysters, egg yolks, and lettuce. The relative effectiveness of each selective plating agar was determined by recovery of Salmonella spp. and enumeration of false-positive and false-negative reactions. Although the new selective plating agars compared favorably with the AOAC/BAM-recom mended agars, they offered no advantage. Incubation of selective enrichment broths at elevated temperatures decreased the numbers of false-positive and falsenegative reactions for all 6 selective plating agars.

1995 ◽  
Vol 78 (2) ◽  
pp. 375-380 ◽  
Author(s):  
Geraldine A June ◽  
Patricia S Sherrod ◽  
Thomas S Hammack ◽  
R Miguel Amaguana ◽  
Wallace H Andrews

Abstract The effectiveness of selenite cystine (SC) broth, tetrathionate (TT) broth, and Rappaport-Vassiliadis (RV) medium for recovery of Salmonella spp. from 8 highly contaminated foods was determined. RV medium prepared from individual ingredients and incubated at 42° and 43°C was compared with 2 commercial (Difco and Oxoid) media incubated at 42°C. Naturally and artificially contaminated foods were tested under 2 protocols. For Protocol 1, each food was preenriched in the appropriate medium. After incubation, serial 10 fold dilutions of the preenriched foods were inoculated into selective enrichment media and incubated at 35°, 42°, or 43°C. Effectiveness of these conditions was evaluated by most probable number determination of Salmonella spp. recovered. Productivity of selective enrichments did not differ significantly with this protocol, except that with Oxoid RV medium the number of Salmonella cells recovered from most of the foods was significantly reduced. For Protocol 2, twenty 25 g test portions from artificially inoculated foods were examined qualitatively for Salmonella spp. The effectiveness of the broth/temperature combinations was determined by the number of positive tests under each condition. RV medium prepared from individual ingredients and TT broth incubated at 43°C yielded significantly more Salmonella-positive tests from frog legs and lettuce than did SC and TT broths incubated at 35°C or commercial RV medium incubated at 42°C. With pork sausage and ground beef, significantly fewer Salmonella-positive tests were found with Oxoid RV medium incubated at 42°C and SC incubated at 35°C than from other selective enrichments. With chicken, fewer Salmonella-positive tests were found from SC and TT broths incubated at 35°C and Oxoid RV medium incubated at 42°C than from other selective enrichments. There were no significant differences among selective enrichments in the recovery of Salmonella spp. from the remaining foods. Overall, RV prepared from individual ingredients and incubated at 42°C yielded the highest number of Salmonella-posWive tests.


1996 ◽  
Vol 59 (11) ◽  
pp. 1158-1163 ◽  
Author(s):  
HUAIZE TIAN ◽  
TAKAHISA MIYAMOTO ◽  
TAKASHI OKABE ◽  
YOICHIRO KURAMITSU ◽  
KEN-ICHI HONJOH ◽  
...  

A rapid-detection method was developed for food-borne dulcitol-positive Salmonella spp. in foods that involves a new preenrichment and selective enrichment system and a sandwich ELISA using two monoclonal antibodies against dulcitol 1-phosphate dehydrogenase. Preenrichment and selective enrichment were in Enterobacteriaceae enrichment mannitol (EEM) broth at 42°C for 6 h and in a new dulcitol-magnesium chloride-pyridinesulfonic acid brilliant green-novobiocin (DMPBN) medium at 42°C for 27 h, respectively. The cells were collected from the selective enrichment culture and suspended in 0.1 ml of 1 N NaOH for 2 min. The solution was neutralized with 0.1 ml of 2 M Tris-HCl buffer (pH 7.5) and the mixture was used as a sample for ELISA. The detection sensitivity of the ELISA was 105 CFU of Salmonella spp. per ml of culture. Competing non-Salmonella organisms in raw food did not interfere with the detection of Salmonella cells even when present at 107: 1 (non-Salmonella: Salmonella ratio) in food. Nonmotile Salmonella gallinarum was detected by the ELISA. The minimum detectable number of initial inoculum of Salmonella typhimurium was 0.69 CFU/25 g of raw chicken after the preenrichment in EEM broth and the selective enrichment in DMPBN medium. The present ELISA method required a total analysis time of 36 h including the preenrichment and selective enrichment periods. The ELISA method was compared with a conventional cultural method for the detection of Salmonella cells in 130 samples of raw foods. Of the samples tested, 16 were Salmonella-positive and 114 samples were negative by both methods. False-positive and false-negative results were not encountered.


1974 ◽  
Vol 37 (7) ◽  
pp. 369-371
Author(s):  
W. C. Ladiges ◽  
J. F. Foster ◽  
W. M. Ganz

A direct fluorescent antibody (FA) test and a polyvalent H agglutination (poly H) test with separate gram-negative (GN) and tetrathionate (TT) broth enrichments were compared with culture methodology in detecting salmonellae from experimentally contaminated ground beef. The FA test had the most false-position reactions (28%, but concurrent culture results reduced these to 7%) and the lowest number of false-negative reactions (4%). The poly H test was more efficient when TT broth was used for selective enrichment. On the basis of fewer false-negative reactions and the greater number of false-positive reactions that were culturally confirmed to be salmonellae, the FA test would appear to be a more efficient screening test in detecting salmonellae from artificially contaminated ground beef held at frozen temperatures.


1996 ◽  
Vol 59 (7) ◽  
pp. 683-687 ◽  
Author(s):  
M. UYTTENDAELE ◽  
R. SCHUKKINK ◽  
B. VAN GEMEN ◽  
J. DEBEVERE

A total of 160 poultry products were examined for the presence of pathogenic campylobacters using the traditional agar isolation method and the nucleic acid amplification system NASBA®, both after a 24-h selective enrichment. Pathogenic campylobacters could be isolated from 92 of 160 (57.5%) samples using agar isolation, among which 79 (49.37%) were identified as Campylobacter jejuni, six (3.75%) as C. coli, five (3.12%) as C. lari, and two (1.25%) as unclassified. The NASBA® assay provides a specific and sensitive method for detection of these campylobacters. A total of 149 samples (93.12%) gave similar results for both the traditional isolation procedure on modified Campylobacter charcoal desoxycholate agar and the NASBA® enzyme-linked gel assay detection system. Two false-negative results were obtained with the agar isolation procedure. Nine false-positive results were reported when the NASBA® system was used. However, the high sensitivity of the NASBA® method and indications that in some cases the traditional isolation procedure failed (abundance of a contaminating noncampylobacter bacteria which grew on the Campylobacter selective media) raises doubt about the true nature of these false-positive results. The NASBA® detection assay offers a rapid and useful analytical method when screening for the presence of pathogenic campylobacters. The complete procedure, including 24 h of selective enrichment, required 32 h.


1999 ◽  
Vol 62 (1) ◽  
pp. 16-21 ◽  
Author(s):  
THOMAS S. HAMMACK ◽  
R. MIGUEL AMAGUAÑA ◽  
GERALDINE A. JUNE ◽  
PATRICIA S. SHERROD ◽  
WALLACE H. ANDREWS

The relative effectiveness of Rappaport-Vassiliadis (RV) medium, selenite cystine (SC) broth, and tetrathionate (TT) broth for the recovery of Salmonella spp. from foods with a low microbial load was determined. RV medium made from its individual ingredients and incubated at 42°C was compared with a commercial preparation of SC broth, incubated at 35°C, and TT broth incubated at 35 and 43°C, for the recovery of Salmonella spp. Twenty-one artificially contaminated food types that included dairy foods, spices, and egg products, as well as other low-microbial-load foods, were analyzed. The foods were inoculated with single Salmonella serovars at target levels ranging from 0.04 to 0.4 CFU/g. No significant differences (P ≤ 0.05) among the selective enrichment broths for the recovery of Salmonella spp. from 18 of the foods were observed. Significantly fewer Salmonella-positive test portions of gelatin, guar gum, and nonfat dry milk were recovered with RV medium than with SC broth incubated at 35°C and TT broth incubated at 35 and 43°C. TT broth incubated at 35°C recovered the greatest number of Salmonella-positive test portions. For the recovery of Salmonella spp. from foods with a low microbial load, it is recommended that TT broth incubated at 35°C and RV medium incubated at 42°C be used.


1993 ◽  
Vol 56 (3) ◽  
pp. 201-204 ◽  
Author(s):  
THOMAS S. HAMMACK ◽  
FELICIA B. SATCHELL ◽  
WALLACE H. ANDREWS ◽  
R. MIGUEL AMAGUANA ◽  
GERALDINE A. JUNE ◽  
...  

A 6-h and a 24-h preenrichment procedure were compared for their ability to recover Salmonella spp. from selected low-moisture dairy foods. The foods were artificially inoculated several days before analysis, and 20 replicate test portions per procedure from each food were examined in each experiment. Samples examined by the 6-h abbreviated procedure were preenriched for 6 h at 35°C in an air incubator or water bath and centrifuged at 4,100 × g for 10 min. Pellets were suspended in tetrathionate broth and incubated for 24 h at 35°C. For the 24-h standard procedure, test portions were preenriched for 24 h at 35°C in an air incubator, subcultured to tetrathionate broth, and incubated for 24 h at 35°C. Selective enrichment broths from both procedures were streaked onto selective agar plates, and presumptive Salmonella isolates were identified by conventional biochemical and serological tests. Recovery of Salmonella spp. from instant nonfat dry milk and dry whole milk was equivalent for both preenrichment procedures. However, the relative effectiveness of the two procedures varied in the recovery of Salmonella spp. from noninstant nonfat dry milk, lactic casein, and rennet casein.


1991 ◽  
Vol 54 (7) ◽  
pp. 492-495 ◽  
Author(s):  
GERALDINE ALLEN ◽  
VERNEAL R. BRUCE ◽  
PATRICIA STEPHENSON ◽  
FELICIA B. SATCHELL ◽  
WALLACE H. ANDREWS

Five high-moisture foods were used to evaluate both the effect of a 6 h, rather than the standard 24 h, selective enrichment incubation period, and the efficiency of Rappaport-Vassiliadis (RV) medium relative to the use of selenite cystine (SC) and tetrathionate (TT) broths for the recovery of Salmonella. Cheese and lettuce were artificially inoculated with a pool of two serotypes, whereas the other foods were naturally contaminated. Significantly higher numbers of Salmonella-positive test portions were obtained at 24 h with the following food and media combinations: cheese (TT and RV media), lettuce (SC, TT, and RV media), raw chicken (RV medium), and pork sausage (SC, TT, and RV media). There were no significant differences between the two incubation periods in recovery of Salmonella from turkey. Overall, more Salmonella-positive test portions were obtained from samples of lettuce, chicken, and pork sausage selectively enriched in RV medium than in SC or TT broths. The results of this study indicate that not all high-moisture foods can be selectively enriched for 6 h without a significant loss in recovery of Salmonella. RV medium was superior to SC and TT broths for recovery of Salmonella from some meats and was at least as productive in its recovery from the other high-moisture foods tested.


2001 ◽  
Vol 64 (5) ◽  
pp. 659-663 ◽  
Author(s):  
THOMAS S. HAMMACK ◽  
R. MIGUEL AMAGUAÑA ◽  
WALLACE H. ANDREWS

The relative effectiveness of three methods for the recovery of Salmonella serovars from orange juice was determined. One method, a modified Bacteriological Analytical Manual (BAM) procedure consisted of preenrichment in lactose broth at 35°C for 24 h, selective enrichment, and selective plating. Another method, a National Centers for Disease Control and Prevention (CDC 1) procedure, consisted of direct enrichment in tetrathionate broth at 35°C for 24 and 48 h, followed by selective plating. The third method (also from CDC and designated CDC 2) consisted of preenrichment in Universal Preenrichment (UP) broth at 35°C for 24 h, selective enrichment, and selective plating. In 10 experiments encompassing five different Salmonella serovars and 200 test portions per broth, the CDC 1 method recovered 141 Salmonella-positive test portions, the BAM method recovered 151, and the CDC 2 method recovered 171. In 2 of the 10 experiments, with two different Salmonella serovars, the BAM recovered significantly fewer (P < 0.05) Salmonella-positive test portions than did the CDC 2 method. On the basis of the above results, the second phase of this study focused on a comparison of the effectiveness of the BAM-recommended lactose broth and the CDC 2-recommended UP broth as preenrichment media for the recovery of Salmonella serovars from pasteurized and unpasteurized orange juice. Subsequent culture treatment of the two preenrichments was identical so that the effect of other variables (e.g., different selective enrichment media, various incubation temperatures, and different selective plating agars) on the relative performance of these two preenrichment media was excluded. In one of nine experiments, with pasteurized orange juice, lactose broth recovered significantly fewer (P < 0.05) Salmonella-positive test portions than did UP broth. For the combined results of the nine pasteurized orange juice experiments (180 test portions per broth), lactose broth recovered 99 Salmonella-positive test portions, and UP broth recovered 116. In three of seven experiments, with unpasteurized orange juice, lactose broth recovered significantly fewer (P < 0.05) Salmonella-positive test portions than did UP broth. For the combined results of the seven unpasteurized orange juice experiments (140 test portions per broth), lactose broth recovered 73 Salmonella-positive test portions, and UP broth recovered 117. For both pasteurized and unpasteurized orange juice, the total number of Salmonella-positive test portions recovered with UP broth was significantly greater than the number recovered with lactose broth. These results indicate that UP broth is a more effective enrichment broth for the recovery of Salmonella from orange juice than is lactose broth.


1983 ◽  
Vol 46 (5) ◽  
pp. 391-399 ◽  
Author(s):  
J.-Y. D'AOUST ◽  
H. J. BECKERS ◽  
M. BOOTHROYD ◽  
A. MATES ◽  
C. R. McKEE ◽  
...  

Twelve laboratories from 7 countries compared the productivity of refrigerated (72 h at 5 to 10°C) preenrichment and enrichment broth cultures with a standard cultural procedure for detection of Salmonella in 466 naturally contaminated low and high moisture foods. Refrigerated preenrichment and enrichment cultures identified 92.5 and 94.2% of contaminated samples, respectively. Variations in the ability of laboratories to successfully recover salmonellae under refrigeration test conditions were notable. Three laboratories found complete agreement between results by the standard and refrigeration test procedures and 5 additional laboratories reported >90% accuracy; lowest recovery rate for combined refrigeration results was 77%. Sensitivity of the refrigeration techniques was markedly greater with low than high moisture foods where the latter contributed all but two of the 62 false-negative results encountered in this study. Ability of individual laboratories to recover Salmonella from refrigerated preenrichment and enrichment broth cultures was not significantly different for given food categories. Productivity of paired enrichment-plating media differed widely with food type. Selective enrichment in tetrathionate brilliant green and plating on bismuth sulfite agar showed greatest sensitivity for isolation of Salmonella in high but not in low moisture foods where productivity of the 4 enrichment-plating conditions used in this study was comparable. Results on recoverability of Salmonella from refrigerated broth cultures concur with findings of an earlier comparative study and strongly support incorporation of this novel approach in standard cultural methods for detection of Salmonella in foods.


1991 ◽  
Vol 54 (3) ◽  
pp. 230-231 ◽  
Author(s):  
PAUL B. VANDERLINDE ◽  
FREDERICK H. GRAU

An ELISA kit (TECRA™) for the detection of Listeria spp. was evaluated for its ability to detect these organisms in naturally contaminated meat and in environmental samples from meat processing plants. Of the 170 samples examined, Listeria monocytogenes and/or L. innocua were detected in 74 by enrichment and selective plating. Testing of the enrichment broths with the ELISA kit detected 72 of these positive samples and gave 2 false-negative and 2 false-positive reactions.


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