Comparison of the Antimicrobial and Sanitizer Resistance of Salmonella Isolates from Chicken Slaughter Processes in Korea

Salmonella is a foodborne pathogen worldwide. Outbreaks of Salmonella are commonly associated with consumption of contaminated foods such as poultry products. Therefore, the objective of this study was to determine the occurrence, biofilm formation, antibiotic resistance, and sanitizer resistance of Salmonella enterica isolated from chicken carcasses. A total of 318 samples were collected from 15 chicken slaughterhouses in 8 provinces of Korea. They were then examined for Salmonella contamination. S. enterica isolates were tested for their susceptibilities to 15 antimicrobials by broth microdilution method. Their biofilm formation ability and resistance to sanitizers were also evaluated. Eighty‐two isolates of S. enterica were obtained from the 318 samples. There were 14 serotypes and 2 untypable isolates. Fifty‐seven (69.5%) isolates were resistant to at least one antibiotic while 30 (36.6%) isolates were resistant to 5 or more antibiotics. Two S. Senftenberg and 3 S. Montevideo isolates exhibited considerable biofilm formation ability (A₆₀₀>0.2) following incubation in Luria‐Bertani (LB) broth for 48 h. Biofilm cell survival and recovery growth assay after sanitization showed that most isolates were highly susceptible to 2.5% lactic acid and 0.1% cetylpyridinium chloride. Therefore, lactic acid and cetylpyridinium chloride might be alternatively or additionally used in addition to chlorine‐based sanitizers that are frequently used to reduce Salmonella contamination of chicken carcasses. Our results provide basic information on the distribution of Salmonella serotypes in chicken slaughterhouses. This study also highlights the necessity to improve farming practices and use antimicrobial agents cautiously. This study also suggests that sanitization during the slaughtering process might be necessary to reduce Salmonella contamination of chicken carcasses.     

Prevalence and Characterization of Salmonella Isolated from Chicken Meat in Turkey

This study was conducted in a Turkish province to investigate the presence of Salmonella spp. in 150 chicken meat samples using 2 phenotyping techniques: classic culture technique (CCT) and immunomagnetic separation (IMS). For the confirmation of the isolates at molecular levels, invA gene was detected in these isolates. The presence of invA, class 1 (Cls1) integrons, and integrase (Int1) genes was demonstrated by PCR assay; and the resistance of the isolated Salmonella spp. strains to antibiotics was determined by disk diffusion test. All the cultural and PCR results were evaluated together; Salmonella spp. were detected in a total of 64 (42.66%) chicken meat samples. Contamination rate was higher in carcasses (53.33%, n = 75) than in meat pieces (32%, n = 75). When results of standard culture were compared with IMS technique, IMS (n = 54) showed a clear superiority over the CCT (n = 38). A very high resistance rate (≥89.28%) to vancomycin, tetracycline, streptomycin, or nalidixic acid was found. Trimethoprim‐sulfamethoxazole resistance was present in 32.14%. Relatively lower incidence of resistance (≤8.33%) to gentamicin, chloramphenicol, ampicillin, and ceftriaxone was observed. Concurrent resistance to at least 4 antibiotics was detected in 92.85% of the isolates. Cls1 integrons and Int1 were positive in 80.95% and 95.23% of the isolates, respectively. However, Int1 alone was detected in 15.47% (n = 13). In conclusion, the high prevalence of Salmonella spp. in chicken meat may pose a potential public health risk, and the presence of antibiotic‐resistant Salmonella spp. isolate together with Cls1 integron and/or integrase might play an important role in horizontal antibiotic gene transfer.     

Effect of carvacrol and thymol on Salmonella spp. biofilms on polypropylene

The present study evaluated the effects of carvacrol and thymol against Salmonella spp. biofilm on polypropylene. The efficacy of the compounds was assessed by quantifying Salmonella spp. cells during and after biofilm formation on polypropylene and performing scanning electron microscopy. During biofilm formation, carvacrol and thymol, at subinhibitory concentrations, reduced bacterial counts about 1–2 log, while established Salmonella spp. biofilms were reduced about 1–5 log by carvacrol and thymol, at MIC or 2× MIC. The greatest reduction in carvacrol‐treated biofilms, about 5 log, was observed with 156 and 312 μg mL^?1 (MIC and 2× MIC) in established Salmonella Typhimurium ATCC 14028 biofilms. Thymol showed the greatest reduction, about 4 log, at 624 μg mL^?1 (2× MIC) against mature Salmonella Enteritidis biofilm. Carvacrol and thymol reduced the number of Salmonella spp. cells on polypropylene, suggesting their potential for the control of Salmonella spp. biofilms.     

Prevalence Analysis and Molecular Characterization of Salmonella at Different Processing Steps in Broiler Slaughter Plants in South Korea

In this study, changes in the prevalence of Salmonella during the processing of broiler chicken carcasses were investigated. A total of 1040 fecal swabs and chicken carcasses samples were collected from 2 processing plants at the 4 stages of broiler processing, which included live birds in slaughter line, postevisceration/prewashing, postwashing/prechilling, and postchilling, respectively. The intraspecific biodiversity of the Salmonella isolates was determined using a DiversiLab automated repetitive sequence‐based PCR (rep‐PCR) system. In both plants, the prevalence of Salmonella increased considerably after evisceration (from 4.6% to 30.8%, P < 0.05) and decreased after washing (from 30.8% to 25.4%, P < 0.05). However, the chilling step had little effect on Salmonella prevalence (from 25.4% to 22.7%, P > 0.05). The most frequent Salmonella serovar in plant A was Infantis (35.8%), followed by Enteritidis (26.2%) and Montevideo (15.0%), while Montevideo (43.6%) and Enteritidis (35.9%) were most prevalent in plant B. A difference in the rep‐PCR banding pattern was found to be related to the processing plant origin and serovar rather than sampling point or sampling day, although there were some exceptional strains.     

Application of magnetic immuno-polymerase chain reaction assay for detection of <Emphasis Type="Italic">Salmonella</Emphasis> spp. in chicken meats

Since contaminated chicken meats have been the principal foodborne source of the contamination of Salmonella to human beings and cultural detection methods are labor-intensive and time-consuming, a study evaluating the performance of the combination of two techniques that are immunomagnetic separation (IMS) and polymerase chain reaction (PCR) for the detection of Salmonella in chicken meats was conducted. The IMS and PCR assay combines selective extraction of Salmonella by specific antibodies with primer-specific (primer pair based on the sequence of invA gene) PCR amplification. Initially chicken meat samples, in which no Salmonella contamination had been determined by using ISO 6579 reference method, were inoculated with Salmonella Enteritidis culture and subsequently the shortest non-selective pre-enrichment time, that had been needed for the detection of approximately 1 or 10 CFU/mL chicken meat levels of target bacteria by magnetic immuno-PCR assay, was found by using 14, 12, 10 and 8-h periods. In conclusion, it was found that magnetic immuno-PCR assay was able to detect 1–10 CFU Salmonella/25 g chicken meat, after only incorporating a non-selective pre-enrichment period of 12 h. Therefore, an overall 16-h (magnetic immuno-PCR assay in conjunction with 12-h non-selective pre-enrichment) magnetic immuno-PCR assay statistically evaluated as sufficient (p = 0.182 > 0.05) for rapid and sensitive detection of approximately 1–10 CFU Salmonella from 25 g chicken meat samples. Accordingly, 16-h magnetic immuno-PCR assay can be promising for routine use in the detection of Salmonella in chicken meat samples, and it consequently may prevent the risk of Salmonella infections in regard to chicken meats.     

A research note: the potential for transfer of Salmonella from irrigation water to tomatoes

The transmission of Salmonella Enteritidis from soil to fruit by contaminated irrigated water was studied using 20 patio tomato plants. In order to track the presence of Salmonella in the soil and plants a luminescent strain transformed with the full luxCDABE gene cassette from Photorhabdus luminescens was used. The tomato plants were irrigated every other day by direct application of water containing Salmonella Enteritidis (10⁵ CFU ml^?1) to the soil. Samples of soil, stem, leaf and fruit were taken weekly and assayed for Salmonella by plating onto Luria Bertani agar containing 50 µg ml^?1 ampicillin. There was a significant difference (P < 0.05) in Salmonella counts from soils sampled during the course of the study. No Salmonella were recovered from the leaf, stem, and fruit samples taken from the tomato plants. This indicates that, under these test conditions, watering with contaminated water directly into the soil does not result in the transmission of Salmonella, and possibly other pathogens, to tomatoes. Copyright © 2004 Society of Chemical Industry     

A new 7-plex PCR assay for simultaneous detection of shiga toxin-producing Escherichia coli O157 and Salmonella Enteritidis in meat products

A 7-plex PCR assay was developed to achieve an effective detection and identification of serotype Enteritidis of Salmonella spp. and shiga toxin-producing type of Escherichia coli O157 in meat products. Six DNA sequences in the invA and sdfI genes of Salmonella Enteritidis as well as rfbE, eae, stx1, and stx2 genes of E. coli O157:H7 were employed to design primers. The multiplex PCR assay could specifically and sensitively detect and identify target pathogens. Applying the assay to meat samples, the multiplex PCR assay was able to simultaneously detect and identify the two pathogens at a sensitivity of three CFU/10 g raw meats after simple 16 h enrichment in buffered peptone water. Further applying in 21 retail meat samples revealed that two samples were positive for non-shiga toxin producing E. coli O157, one sample was positive for Stx2 producing E. coli O157 and five samples were positive for Salmonella enterica Enteritidis. Taken together, the 7-plex PCR assay is a rapid and reliable method for effectively screening single or multiple pathogens occurrences in various meat products, and could also identify the Salmonella enterica Enteritidis from all Salmonella spp. and shiga toxin producing type from all E. coli strains. Considering as a non expensive screening tool, the multiplex PCR assay has a great potential in complement for food stuff analysis by conventional microbiological tests.     

Salmonella Anatum,S. Infantis and S. Schwarzengrund in Brazilian Cheeses: Occurrence and antibiotic resistance profiles

Salmonella is one of the major causative agents of foodborne infections. In this study, 225 samples of different types of cheeses produced by the Brazilian dairy industry were analysed. Samples were submitted to a Salmonella spp. investigation using conventional microbiology, multiplex polymerase chain reaction (multiplex PCR) and the disc‐diffusion method. The occurrence of Salmonella was 1.33% (3/225); two strains (S. Infantis and S. Schwarzengrund) were detected in Prato cheese, and one strain (S. Anatum) was detected in Mozzarella cheese. The three strains showed resistance to the antifolate pathway inhibitors trimethoprim and sulphonamide. Therefore, Salmonella spp. in cheese samples indicates a threat to consumer public health.     

Salmonella spp. contamination in fresh pork and chicken sausages marketed in Niterói and Rio de Janeiro,Brazil

Fresh sausages are one of the most popular meat products consumed in Brazil. However, the use of contaminated raw material, unhygienic handling during processing, and inadequate storage could be hazardous to consumer’s health, since fresh-sausage processing does not include heat treatment and the product may carry several human pathogens such as Salmonella spp.. In this study, a total of 80 fresh sausages prepared with pork and chicken meat were evaluated regarding Salmonella spp. presence by PCR after selective enrichment and bacteriological culture. Twenty-one sausage samples (27 %) proved to be contaminated with Salmonella spp.. Sixteen (76 %) contaminated samples were detected by PCR combined with enrichment broth culturing, whereas conventional microbiological screening detected only 8 (38 %). In this study, evidence was raised regarding packing conditions of fresh sausages, since it seems that there is no significant difference in Salmonella spp. contamination among samples in their original packing (23 %), wrapped in plastic or unpacked (27 %). Similarly, meat composition seems not to have a determinant influence in contamination, since 20 % of the chicken and 29 % of the pork sausages were contaminated. The contamination rate of Tuscan-type sausages was 37 %, while contamination was found in 21 % of ham sausages, suggesting that the cut of pork did not have a determinant influence in the contamination by Salmonella in fresh sausages. The results shown here indicate that fresh sausages marketed in Niterói and Rio de Janeiro may be contaminated with Salmonella spp., requiring more attention by sanitary authorities in order to assure the safety of this food.     

Validation of Real-Time PCR and Enzyme-Linked Fluorescent Assay-Based Methods for Detection of Salmonella spp. in Chicken Feces Samples

A comparative study of enzyme-linked fluorescent assay (ELFA)-based methods and real-time polymerase chain reaction (PCR)-based methods using three and two different sample preparation protocols, respectively, and the standard culture-based method EN ISO 6579:2002/Amd1:2007, for the detection of Salmonella spp. in chicken feces, was performed on 20 artificially and 68 naturally contaminated chicken feces. Selectivity, relative specificity, relative accuracy, relative sensitivity, and relative detection level were determined. According to criteria established in the methods comparison study included in EN ISO 16140:2003 for validation of alternative microbiological methods, the ELFA-based methods (V1 and V2) as well as a real-time PCR method (PCR2) were comparable to the reference method for the detection of Salmonella in chicken feces. They provided results in 48 h and presented a high sensitivity (97% for all of them). The three methods showed a relative specificity of 94%, V1 being the method which presented the highest relative accuracy (96%). While detection level for V2 and reference method was between 3 and 13 CFU/25 g, PCR2 method was able to detect down to 3 CFU/25 g. In conclusion, both the real-time PCR and the ELFA-based assays can be used as rapid and user-friendly screening methods for detection of Salmonella spp. in chicken feces.     

A new 7-plex PCR assay for simultaneous detection of shiga toxin-producing Escherichia coli O157 and Salmonella Enteritidis in meat products

A 7-plex PCR assay was developed to achieve an effective detection and identification of serotype Enteritidis of Salmonella spp. and shiga toxin-producing type of Escherichia coli O157 in meat products. Six DNA sequences in the invA and sdfI genes of Salmonella Enteritidis as well as rfbE, eae, stx1, and stx2 genes of E. coli O157:H7 were employed to design primers. The multiplex PCR assay could specifically and sensitively detect and identify target pathogens. Applying the assay to meat samples, the multiplex PCR assay was able to simultaneously detect and identify the two pathogens at a sensitivity of three CFU/10?g raw meats after simple 16?h enrichment in buffered peptone water. Further applying in 21 retail meat samples revealed that two samples were positive for non-shiga toxin producing E. coli O157, one sample was positive for Stx2 producing E. coli O157 and five samples were positive for Salmonella enterica Enteritidis. Taken together, the 7-plex PCR assay is a rapid and reliable method for effectively screening single or multiple pathogens occurrences in various meat products, and could also identify the Salmonella enterica Enteritidis from all Salmonella spp. and shiga toxin producing type from all E. coli strains. Considering as a non expensive screening tool, the multiplex PCR assay has a great potential in complement for food stuff analysis by conventional microbiological tests.     

Synergistic Reduction of Salmonella in a Model Raw Chicken Media using a Combined Thermal and Acidified Organic Acid Salt Intervention Treatment

ABSTRACT: Salmonella-contaminated poultry products are considered major contributors to foodborne illness. The anti-Salmonella activity of organic acid salts has been studied in food products and poultry feed but rarely in combination with nonchemical treatments. Here, we investigated the combination of acidified organic acid salt solutions with thermal treatment as an effective Salmonella intervention applicable in poultry carcass processing. A model raw chicken media was used to propagate Salmonella prior to the intervention treatment. Salmonella Typhimurium strains LT2 and ATCC nr 14028 grew similarly in the model raw chicken media at 37 and 42 °C, reaching stationary phase 24 h after inoculation. Four log₁₀CFU of either Salmonella Typhimurium strain at stationary phase was exposed to 2.5% organic acid salt solutions (at pH 4) for 1 min at 55 °C. All organic acid salt treatments yielded significant Salmonella Typhimurium reductions, ranging from 1 log (sodium acetate) to almost 4 logs (sodium butyrate). Exposure to pH 4 water at 55 °C or the organic acid salt solutions at room temperature had no effect. The combined thermal and acidified organic acid salt intervention produced a significant, synergistic reduction of Salmonella Typhimurium and may represent an effective method for decontamination of poultry carcasses during processing.     

Thermal inactivation of Salmonella spp. in ground chicken breast or thigh meat

Thermal inactivation of a four‐strain mixture of Salmonella spp. was determined in chicken breast and thigh meat. Inoculated meat was packaged in bags and then completely immersed in a circulating water bath and held at 55, 57.5, 60 and 62.5 °C for pre‐determined lengths of time. When the surviving bacteria were enumerated on tryptic soya agar supplemented with 0.6% yeast extract and 1% pyruvate (non‐selective medium), D‐values (time to inactivate 90% of bacteria) in chicken breast meat were 6.08, 4.77, 3.00 and 0.66 min at 55, 57.5, 60 and 62.5 °C, respectively; the values in thigh meat ranged from 11.48 min at 55 °C to 0.84 min at 62.5 °C. As expected, the measured heat resistance was lower when the recovery medium was selective (xylose lysine deoxycholate agar). Thermal death time values from this study will assist food processors in designing the HACCP plans to effectively eliminate Salmonella spp. in cooked chicken breast and thigh meat.     

Prevalence and antimicrobial resistance of Salmonella serotypes isolated from poultry in Spain: comparison between 1993 and 2006

A total of 226 chicken samples (carcasses, legs, wings, necks and breasts) were obtained (73 in 1993 and 153 in 2006) from 10 retail outlets in North-Western Spain and screened for the presence of Salmonella. Isolates were subjected to serotyping, phage typing (Salmonella Enteritidis and Salmonella Typhimurium) and antimicrobial susceptibility testing (15 antimicrobials; disk diffusion method). Salmonella was detected in 40 (55%) samples in 1993 and 19 (12.4%) in 2006 (P < 0.001). The serotypes (S. Enteritidis, Salmonella Poona, Salmonella Infantis, Salmonella Newport and S. Typhimurium) and phage types (1, 4, 14b and 35 in the case of S. Enteritidis and 193 for S. Typhimurium) detected are among the main types responsible for human salmonellosis in Spain. All strains were multi-resistant (resistant to 3-13 antimicrobials). The average number of resistances per strain increased (P < 0.05) from 3.98 in 1993 to 5.00 in 2006. An increase in the incidence of resistance was observed between 1993 and 2006 for cephalothin (P < 0.01), enrofloxacin (P < 0.001) and tetracycline (P < 0.01). The decreases in the prevalence of Salmonella between 1993 and 2006 suggest that the mandatory measures introduced over the last decade in the European Union to reduce the incidence of Salmonella in poultry have apparently been successful. However, the increase in antibiotic resistance rates is of concern and constitutes a threat to public health. Because the data in this study demonstrated that chicken in North-Western Spain is a potential source of antibiotic-resistant Salmonella strains, the need of consumer education on good sanitary practices is highlighted.     

Occurrence of salmonellae in retail chicken carcasses and their products in Spain

This study examined the incidence of Salmonella in Spanish poultry products. Samples included chicken carcasses, chicken parts (wings, legs and giblets-livers and hearts) and processed chicken products (red sausages, white sausages and hamburgers). The average detection rate was 49%, with the highest (55%) in chicken carcasses (skin) and the lowest (20%) in hamburgers. The chicken carcasses purchased in supermarkets were more contaminated (75%) than those from poulterers shops (25%). Salmonella Enteritidis, S. Poona, S. Paratyphi B and S. Worthington were isolated in 34.3%, 11.4%, 2.8% and 1.4% of the samples, respectively. One (1.4%) red sausage sample harboured two serotypes (S. Enteritidis and S. Worthington). This fact emphasizes the usefulness of subtyping several Salmonella isolates from the same sample in epidemiological studies.     

Loop-Mediated Isothermal Amplification (LAMP) for the Rapid and Sensitive Detection of Salmonella Typhimurium from Pork

ABSTRACT: Loop-mediated isothermal amplification (LAMP) is a novel molecular detection method that is more rapid and simpler than PCR. Products can be detected by turbidity using one temperature without the need for expensive PCR equipment. Our objective was to sensitively detect Salmonella Typhimurium from pork products within 1 d using the LAMP assay. Pork chop and pork sausage samples (25 g) were inoculated with high (10⁸ to 10⁶ CFU) and low (10⁵ to 10⁰ CFU) inocula of S. Typhimurium. Serial dilutions in phosphate buffered saline were plated on XLT4 agar either immediately or after selective preenrichment in tetrathionate broth (225 mL) for 10-h at 37 °C. Nucleic acid was extracted using the TRIzol^® method from 1-mL samples. The LAMP assay using 6 specific invA gene primers and Bst DNA polymerase reaction mix was carried out at 62 °C for 90 min in a waterbath. Turbid products were detected visually and by agarose gel electrophoresis. Improved Salmonella detection at 10² CFU/25 g for both pork chop and sausage was obtained after 10-h enrichment and 10⁶ CFU/25 g without enrichment for both products. This assay can detect Salmonella from pork within 1 d, significantly faster than traditional methods that take >5 d. This method shows tremendous potential for routine diagnostics and monitoring of Salmonella by the pork industry. Practical Application: The novel loop-mediated isothermal amplification (LAMP) assay is a rapid, specific, and sensitive method that has potential application for routine diagnostics of Salmonella from pork products. The isothermal method does not require expensive equipment such as a PCR thermocylcer but only a simple waterbath for amplification within 90 min. Detection is even simpler by visual eye or turbidimeters that are less expensive than fluorescent spectrophotometers or real-time PCR machines. All these advantages make it a practical approach for routine use by processing industries to rapidly detect Salmonella in their environment and to implement appropriate control strategies. To improve detection sensitivities, preenrichment followed by selective enrichment may be necessary. Even so, the entire assay can be completed at the most within two 8-h working shifts.     

Multiplex PCR coupled with propidium monoazide for the detection of viable Cronobacter sakazakii,Bacillus cereus,and Salmonella spp. in milk and milk products

Cronobacter sakazakii, Bacillus cereus, and Salmonella spp. are common food-borne pathogens. The aim of this study was to develop a sensitive, specific, and rapid method for the simultaneous detection of these 3 pathogens in milk and milk products. Three specific primers were designed based on ompA, invA, and cesB of C. sakazakii, Salmonella spp. and B. cereus, respectively, for use in a multiplex PCR (mPCR). To eliminate false-positive results, cells were pretreated with propidium monoazide (PMA) for the selective elimination of the genomic DNA of dead cells. An internal amplification control was applied as an indicator of false-negative results from the interference of inhibitors in the food matrix. Results showed that, in pure culture, the limits of detection of the assay for C. sakazakii, Salmonella Enteritidis, and B. cereus were 9.5 × 10⁴, 7.4 × 10², and 7.5 × 10² cfu/mL, respectively. Moreover, 8 cfu/mL of viable B. cereus cells were detected after 5 h of enrichment, and 9 cfu/mL of viable C. sakazakii and 7 cfu/mL of Salmonella Enteritidis were detected after 7 h of enrichment in spiked pure milk, walnut peanut milk, and whole-wheat milk. To validate the PMA-mPCR assay, the PMA-mPCR assay and the traditional culture method were performed to detect the 3 bacterial strains in 1,165 milk product samples. The PMA-mPCR assay obtained the same results as the culture-based method. Results demonstrated that the PMA-mPCR assay has excellent sensitivity and specificity for the simultaneous detection of viable C. sakazakii, Salmonella Enteritidis, and B. cereus in milk and milk products.     

Cell Surface Display of Salmonella Outer Membrane Protein A on Lactobacillus salivarius: A First Step Towards Food-Grade Live Vaccine Against Salmonella Infections

As one of the most important pathogens in both humans and animals, Salmonella is a significant health problem, especially in the developing countries. Outer membrane protein A (OmpA) is among the major outer membrane proteins of Salmonella with strong immunogenicity. Vaccination is an effective tool for the prevention of Salmonella infections. Lactic acid bacteria are the normal flora of gastrointestinal (GI) tract. In this research, lactobacilli from different parts of chicken GI tract were isolated and evaluated for bile salts and acidic pH tolerance. One strain isolated from the small intestine was identified as Lactobacillus salivarius by carbohydrate fermentation test and 16SrRNA partial sequencing. To address the need for safe oral vaccine to control Salmonella infection, we engineered this natural chicken isolate of L. salivarius to express OmpA under promotion of Lactate dehydrogenase (ldh) promoter. The surface displayed OmpA reacted with anti-OmpA in ELISA, Immunoblotting and flow cytometry assays, suggesting that the expressed protein adopted a native conformation. Strong agglutination reaction of the recombinant lactobacilli with sera of the naturally Salmonella infected chickens showed the multivalency and overexpression of the OmpA on the surface of L. salivarius. Experimental infection assay confirmed that the engineered commensal bacteria could be an attractive candidate as food-grade live vaccine against Salmonella infections.     

Detection of Salmonella spp. in Oysters Using Polymerase Chain Reactions (PCR) and Gene Probes

The polymerase chain reaction (PCR) DNA amplification method was used to identify oligonucleotide primers from a target gene, hns, to specifically detect SulmonelIu spp. in contaminated oysters. The primers for PCR amplification and a hybridizing oligonucleotide probe to detect the amplified DNA were specific for all Salmonella spp. tested. Modification of a procedure for extracting DNA from oyster tissue matrices followed by PCR amplification, and coupled with gene-probe hybridization detected <40 cells of seeded or naturally occurring Sulmonella spp./g of oyster meat samples without any enrichment step. The detection of SaZmonella spp. was reliable, sensitive, and required considerably less time than conventional microbiological culture methods.