Salmonella survival and differential expression of fatty acid biosynthesis-associated genes in a low-water-activity food.

UNLABELLED: The purpose of this study was to investigate the difference in expression of fatty acid biosynthesis genes and survival of different serotypes of Salmonella when incubated in a low-water-activity (aw ) food over a 14-day period. Stationary cells of five strains of Salmonella enterica belonging to 3 different serovars (Typhimurium ATCC 2486, Enteritidis H4267, Tennessee ARI-33, Tennessee S13952 and Tennessee K4643) were inoculated into granular sugar (aW   = 0·50) and held aerobically over a 14-day period at 25°C. Survival was determined by enumerating colonies on TSA and XLT-4 plates at 0, 1, 3, 5, 7 and 14 days. Correspondingly, gene expression was evaluated for three selected genes involved in fatty acid biosynthesis and modification (fabA, fabD and cfa). After 14 days of incubation, the population was reduced from 2·29 to 3·36 log for all five strains. Salmonella Tennessee ARI-33 and Salm. Tennessee K4643 displayed greater survival than Salm. Typhimurium and Salm. Enteritidis. The increased expression of the cfa gene (involved in cyclopropane fatty acid biosynthesis) over 14 days was found associated with strains with a lower survival rate. The fabA gene (involved in unsaturated fatty acid biosynthesis) was observed up-regulated for all strains for at least one sampling time and for Salm. Tennessee ARI-33 for all time points tested, suggesting its potential role in enhancing Salmonella survival in low aw foods. SIGNIFICANCE AND IMPACT OF THE STUDY: Numerous outbreaks of salmonellosis associated with low-water-activity foods have been reported. Therefore, the adaptive mechanisms utilized by Salmonella to survive in low-water-activity foods for prolonged periods of time need to be better understood. The results in this study showed that low-water-activity environments increase expression of gene fabA, which is involved in unsaturated fatty acid biosynthesis of Salmonella, while the increased expression of cfa, associated with cyclopropane fatty acid synthesis, was associated with decreased survival over 14 days.

Occurrence of Escherichia coli O157:H7 in diverse farm environments.

In the United States, foodborne outbreaks of Escherichia coli O157:H7 illness have often been linked to the consumption of contaminated, undercooked ground beef. However, the occurrence of E. coli O157:H7 has also been reported in other farm animals. The objective of this study was to evaluate the occurrence of E. coli O157:H7 on diverse farm types and from a variety of farm samples. Rectal swabs (n=1686) and environmental samples (n=576) were collected from 16 farms in five states over 24 months and analyzed for the presence of E. coli O157:H7. Overall, E. coli O157:H7 was found in 3.6% of beef cattle, 3.4% of dairy cattle, 0.9% of chicken, 7.5% of turkey, and 8.9% of swine samples. The pathogen was isolated sporadically from each of the environmental sample types. Of particular concern was the isolation of E. coli O157:H7 from fresh feed samples, indicating a potential vector for transmission. The data from this study indicate a high occurrence of E. coli O157:H7 on swine and turkey farms. This unexpected result suggests that more research on the occurrence of E. coli O157:H7 on these types of farms is required in order to better understand potential reservoirs of pathogenic E. coli.

Evaluation of methods for recovery of Salmonella from dairy cattle, poultry, and swine farms.

Current official methods for detection and isolation of Salmonella are mostly designed for foods. The objective of this study was to determine optimal methods for detection and isolation of Salmonella from animal and environmental samples of dairy, poultry, and swine farms. Preenrichment in lactose broth versus direct enrichment (no preenrichment) prior to selective enrichment in Rappaport-Vassiliadis, selenite cystine, and tetrathionate incubated at 35 and 42 degrees C and in four differential/selective plating media (brilliant green, bismuth sulfite, Hektoen enteric, and xylose-lysine-tergitol 4 agar base) were evaluated for their ability to recover Salmonella from artificially contaminated samples. The effects of pH adjustments to samples on Salmonella recovery were determined. A pH adjustment of the enrichment broth to 6.8 +/- 0.2 after addition of samples significantly improved recovery of Salmonella. The most effective medium combinations for isolation of Salmonella from farm samples depended on the type of samples. Generalizations of protocols for recovery of Salmonella from farm samples might result in poor recovery, increased recovery time, and increased sample processing costs.

Comparison of dichloran 18% glycerol (DG18) agar with general purpose mycological media for enumerating food spoilage yeasts.

Dichloran 18% glycerol (DG18) agar was originally developed to enumerate xerophilic foodborne moulds. However, some laboratories are using DG18 agar as a general medium to enumerate foodborne moulds and yeasts. A collaborative study, with the participation of seven laboratories, was undertaken to compare DG18 agar with dichloran rose bengal chloramphenicol (DRBC) agar, tryptone glucose yeast extract chloramphenicol (TGYC) agar, and plate count agar supplemented with chloramphenicol (PCAC) for enumerating 14 species of common food spoilage yeasts. Comparison of the mean values of populations of all yeasts recovered on each medium revealed no significant differences among DRBC agar, PCAC, and TGYC agar, while each of these media supported the development of significantly (P < or = 0.05) higher numbers of colonies than DG18 agar. However, differences were only 0.08 to 0.10 log10 cfu/ml, making the practical significance questionable. The overall coefficient of variation (CV) for within laboratory repeatability was 1.71%, while the CV for reproducibility of counts obtained among laboratories was 6.96%. Compared to DRBC agar, TGYC agar, and PCAC, yeast colonies were smaller on DG18 agar. Growth of Brettanomyces anomalus, Cryptococcus albidus, and Rhodotorula mucilaginosa was particularly retarded or inhibited on DG18 agar. Based on the performance of media in supporting colony development and ease of counting colonies, the use of DG18 agar as a general enumeration medium for foodborne yeasts cannot be recommended.

Antimicrobial activity of essential oils from plants against selected pathogenic and saprophytic microorganisms.

The beneficial health effects of extracts from many types of plants that are used as seasoning agents in foods and beverages have been claimed for centuries. The purpose of this study was to examine the effectiveness of selected herb and spice essential oils for control of growth and survival of microorganisms. Inhibition of growth was tested by the paper disc agar diffusion method. Antibiotic susceptibility discs were used as control. Minimum lethal concentration (MLC) was determined by the tube dilution method. Essential oils from anise, angelica, basil, carrot, celery, cardamom, coriander, dill weed, fennel, oregano, parsley, and rosemary were evaluated. Inhibition ranged from complete with oregano to no inhibition with carrot oil for each of the test strains that included: Listeria monocytogenes, Staphylococcus aureus, Escherichia coli O:157:H7, Yersinia enterocolitica, Pseudomonas aeruginosa, Lactobacillus plantarum, Aspergillus niger, Geotrichum, and Rhodotorula. Oregano essential oil showed the greatest inhibition (zone, > or = 70 to 80 mm) (MLC, approximately 8 ppm). Coriander and basil were also highly inhibitory (MLC, approximately 25 to 50 ppm) to E. coli O:157:H7 and to the other bacteria and fungi tested. Anise oil was not particularly inhibitory to bacteria (inhibition zone, approximately 25 mm); however, anise oil was highly inhibitory to molds. Because some of the herbal and spice essential oils are highly inhibitory to selected pathogenic and spoilage microorganisms, they may provide alternatives and supplements to conventional antimicrobial additives in foods.

Influence of modified atmospheric storage, lactic acid, and NaCl on survival of sublethally heat-injured Listeria monocytogenes.

The effect of package atmosphere on survival of uninjured and sublethally heat-injured Listeria monocytogenes, inoculated onto tryptose phosphate agar containing 0.85% lactic acid and 2% NaCl (TPALAS) was investigated. Inoculated TPALAS plates were packaged in air, 100% N2 (N2), 30% CO2-70% N2 (CO2-N2), and vacuum and stored at 4 and 20 degrees C for up to 31 days. Recovery of L. monocytogenes from TPALAS was influenced by the injury status (i.e., injured and uninjured) of the inoculum, storage atmosphere (air, N2, CO2-N2, and vacuum), storage temperature (4 and 20 degrees C), and recovery media [tryptose phosphate agar (TPA) and modified Oxford agar (MOX)] (P <0.05). Overall, storage at 4 degrees C supported greater survival than storage at 20 degrees C (P< 0.05). Uninjured L. monocytogenes stored at 4 degrees C was recovered on TPA better than sublethally heat-injured L. monocytogenes stored at 40 degrees C (P < 0.05). Recovery of sublethally heat-injured L. monocytogenes stored at 4 degrees C followed the order N2 > CO2-N2 > air > vacuum (P < 0.05), whereas recovery of uninjured L. monocyrogenes stored at 4 degrees C followed the order N2 > CO2-N2 > vacuum > air (P < 0.05). Air and vacuum atmospheres supported greater survival of uninjured and heat-injured L. monocytogenes than N2 and CO2-N2 atmospheres at 20 degrees C (P < 0.05). Recovery of sublethally heat-injured L. monocytogenes stored at 20 degrees C followed the order vacuum > air> CO2-N2 = N2 (P <0.05), whereas recovery of uninjured L. monocytogenes stored at 20 degrees C followed the order vacuum > air> CO2-N2 > N2 (P<0.05). Uninjured L. monocytogenes stored under N2 at 4 degrees C was recovered best, whereas sublethally heat-injured L. monocytogenes stored under N2 at 20 degrees C was recovered poorest (P < 0.05). Factors such as package atmosphere and storage temperature, involved in the production, storage, and distribution of fermented foods must be thoroughly evaluated when determining strategies for control and detection of L. monocytogenes in such products.

Growth and survival of uninjured and sublethally heat-injured Escherichia coli O157:H7 on beef extract medium as influenced by package atmosphere and storage temperature.

The effect of atmospheric composition and storage temperature on growth and survival of uninjured and sublethally heat-injured Escherichia coli O157:H7, inoculated onto brain heart infusion agar containing 0.3% beef extract (BEM), was determined. BEM plates were packaged in barrier bags in air, 100% CO2, 100% N2, 20% CO2: 80% N2, and vacuum and were stored at 4, 10, and 37 degrees C for up to 20 days. Package atmosphere and inoculum status (i.e., uninjured or heat-injured) influenced (P < 0.01) growth and survival of E. coli O157:H7 stored at all test temperatures. Growth of heat-injured E. coli O157:H7 was slower (P < 0.01) than uninjured E. coli O157:H7 stored at 37 degrees C. At 37 degrees C, uninjured E. coli O157:H7 reached stationary phase growth earlier than heat-injured populations. Uninjured E. coli O157:H7 grew during 10 days of storage at 10 degrees C, while heat-injured populations declined during 20 days of storage at 10 degrees C. Uninjured E. coli O157:H7 stored at 10 degrees C reached stationary phase growth within approximately 10 days in all packaging atmospheres except CO2. Populations of uninjured and heat-injured E. coli O157:H7 declined throughout storage for 20 days at 4 degrees C. Survival of uninjured populations stored at 4 degrees C, as well as heat-injured populations stored at 4 and 10 degrees C, was enhanced in CO2 atmosphere. Survival of heat-injured E. coli O157:H7 at 4 and 10 degrees C was not different (P > 0.05). Uninjured and heat-injured E. coli O157:H7 are able to survive at low temperatures in the modified atmospheres used in this study.

Fate of Escherichia coli O157:H7 in ground apples used in cider production.

Survival of Escherichia coli O157:H7 in ground Golden Delicious, Red Delicious, Rome, and Winesap apples stored at 4, 10, and 25 degrees C was determined. E. coli O157:H7 populations were monitored for up to 18 days (4 degrees C), 12 days (10 degrees C), and 5 days (25 degrees C), when mold contamination became visible. At 25 degrees C, Red Delicious apples supported survival of E. coli O157:H7 better (P < 0.05) than the other cultivars, followed by Golden Delicious and Rome apples, which were not statistically different (P > 0.05). Winesap apples were the least favorable (P < 0.05) for survival of E. coli O157:H7 at 25 degrees C. E. coli O157:H7 was recovered at similar rates from Golden Delicious and Red Delicious apples, (P > 0.05), but pathogen populations increased in both cultivars (P < 0.05) during storage at 25 degrees C. At 10 degrees C, survival of E. coli O157:H7 was poorest (P < 0.05) in ground Red Delicious apples, while there was no significant difference in survival of E. coli O157:H7 among ground Golden Delicious, Rome, or Winesap cultivars (P > 0.05). When stored at 4 degrees C, Golden Delicious and Rome apples were not statistically different in supporting survival of the pathogen (P > 0.05). In general, apple pH increased during storage and was associated with mold growth. Results of this investigation indicate that there is no trend toward a particular apple cultivar supporting survival of E. coli O157:H7. However, variation in apple pH during storage can negatively or positively influence E. coli O157:H7 survival at 25 degrees C.

Effects of a direct-fed yeast culture on enteric microbial populations, fermentation acids, and performance of weanling pigs.

In three replicate trials, a total of 36 pigs that had been cannulated at the terminal ileum were used to determine the effects of a Saccharomyces cerevisiae culture in a phase feeding program (phase I was d 0 to 7 and phase II was d 8 to 21) on performance, ileal microflora, and short-chain fatty acids in weanling pigs. Pigs were cannulated at approximately 12 d of age, weaned at 17 d of age, and randomly assigned to one of three treatments: 1) a pelleted phase feeding program, 2) a similar program with the inclusion of a live S. cerevisiae culture (1 g/ kg), and 3) a nonpelleted feeding program otherwise similar to program 2. Ileal samples were collected at 17, 20, 24, 27, 31, 34, and 38 d of age, and samples were analyzed for total E. coli, streptococci, lactobacilli, yeast, short-chain fatty acids, pH, and dry matter. Performance data were also collected. At 41 d of age, pigs were killed and digesta were collected from various regions of the gastrointestinal tract. Total intake was less for pigs fed the control diet than for pigs fed the yeast diets, and overall gains tended to be greater for pigs fed diets including yeast. Treatment differences were not observed for ileal microflora or short-chain fatty acids in samples obtained from cannulas or from the various sites of the gastrointestinal tract. Inclusion of a live yeast culture in weanling pig diets affected intake and performance but did not alter tested intestinal microflora or net concentrations of fermentation products.

Influence of growth temperature on inactivation and injury of Escherichia coli O157:H7 by heat, acid, and freezing.

The influence of growth temperature on heat-, lactic acid-, and freeze-induced inactivation and injury of Escherichia coli O157:H7 in 0.1% peptone water was investigated. Three strains of E. coli O157:H7 isolated respectively from salami, apple cider, and ground beef were evaluated. Growth of strains at 10 degrees C compared with growth at 37 degrees C had a significant impact on reducing (P < 0.01) D values obtained for heating (DH value), acid exposure (DA value), with the exception of the cider strain stored in lactic acid solutions. When strains were cultivated at 10 and 37 degrees C and heated at 54 and 56 degrees C, the salami strain possessed the highest (P < 0.01) DH values (5.9 to 59.7 min). When grown at 10 degrees C, the beef strain had the lowest (P < 0.01) DH values after heating at 52, 54, and 56 degrees C (11.2, 4.1 and 2.5 min, respectively). The salami strain grown at 10 degrees C had the highest (P < 0.01) DA values in all concentrations of lactic acid. When grown at 37 degrees C, the salami strain had the highest (P < 0.01) DA values after storage in 0.1 and 0.25% lactic acid, while DA values for the salami and beef strains did not differ (P > 0.05) when stored in 0.5% lactic acid. Portions of strain populations were sublethally injured by heat and lactic acid treatments, as evidenced by the inability of injured organisms to form colonies on tryptone soy agar containing 2% NaCl. Strains cultured at 10 degrees C were more susceptible to sublethal heat injury than the strains cultured at 37 degrees C. Storage of test strains at -20 degrees C for 7 months resulted in a 4- to 6-log CFU/ml reduction in viable population, but induced only minimal sublethal injury. After 5 months at -20 degrees C, strains cultured at 10 degrees C were more sensitive to freeze inactivation than strains cultured at 37 degrees C. When grown at 10 and cultured at 37 degrees C. When grown at 10 and 37 degrees C and stored at -20 degrees C for 7 months, the cider strain possessed higher (P < 0.01) DF values than the beef and salami strains.

Changes in fatty acid composition of various lipid components of Zygosaccharomyces rouxii as influenced by solutes, potassium sorbate and incubation temperature.

The effects of water activity (aw), solutes, potassium sorbate and incubation temperature on the fatty acid methyl ester composition of the total extracted lipid, triacylglycerol, phospholipid, free fatty acid and steryl ester components of Zygosaccharomyces rouxii were determined. Oleic (C18:1) and linoleic (C18:2) acids were the major components of the total fatty acid fraction, regardless of culturing conditions. Cells grown at 35 degrees C contained a significantly higher percentage of C18:1 and a lower percentage of C18:2 than did cells grown at 21 degrees C. At both temperatures, cells cultured in low-aw (0.93) media contained a significantly higher percentage of C18:1, with a corresponding decrease in C18:2 content than did cells grown at aw 0.99, regardless of the addition of glucose, sucrose, or NaCl to suppress the aw. At 21 degrees C, cells cultured in high-aw (0.99) media supplemented with 300 micrograms sorbate per ml contained higher percentages of C18:1 than did cells cultured in media not supplemented with sorbate. Cells grown at 35 degrees C in low-aw media supplemented with sorbate contained substantially more C18:1 than did cells cultured without sorbate.

Effects of potassium sorbate on growth patterns, morphology, and heat resistance of Zygosaccharomyces rouxii at reduced water activity.

A study was made of the effects of potassium sorbate on growth, morphology, and heat sensitivity of an osmotolerant yeast, Zygosaccharomyces rouxii, grown in media (water activity (aw) 0.93) supplemented with glucose and sucrose. Growth patterns of Z. rouxii in YM broth supplemented with glucose (YMBG) and sucrose (YMBS) were similar, although increased potassium sorbate concentration in both media resulted in decreased growth rates. Growth in YMBS containing potassium sorbate was not as prolific as that in YMBG containing potassium sorbate. Inhibition of growth was indicated by decreased absorbance (at 600 nm) of cells grown in YMBS and in YMBG and YMBS supplemented with potassium sorbate at 600 or 1000 micrograms/mL. Slight decreases in cell size and alteration of cellular morphology were associated with increased potassium sorbate concentration. Plasmolysis increased as potassium sorbate concentration was elevated in YMBS but not in YMBG. Tolerance of Z. rouxii to potassium sorbate was enhanced by previous adaptation of cells in media with elevated potassium sorbate concentrations. Heat resistance of cells unadapted to potassium sorbate showed little or no increase regardless of culture age, but increased substantially in cells grown in media containing potassium sorbate, particularly YMBS.

Interactive effects of solutes, potassium sorbate and incubation temperature on growth, heat resistance and tolerance to freezing of Zygosaccharomyces rouxii.

The interactive effects of solutes, potassium sorbate and incubation temperature on growth, heat resistance and tolerance to freezing of Zygosaccharomyces rouxii were investigated. Growth rates in media supplemented with glucose, sucrose or NaCl to aw 0.93 were more rapid than in unsupplemented media (aw 0.99). Although growth in unsupplemented medium was lower at 35 degrees C, incubation at 21 degrees C or 35 degrees C had little effect on growth in media supplemented with glucose and sucrose. The addition of 300 micrograms potassium sorbate/ml to media resulted in reduced growth rates, particularly at 35 degrees C. Heat resistance of Z. rouxii was substantially greater in cultures previously incubated at 35 degrees C than in cultures incubated at 21 degrees C in media both with and without 300 micrograms potassium sorbate/ml. Zygosaccharomyces rouxii was tolerant to freezing at -18 degrees C for up to 120 d in all test media supplemented with glucose, sucrose or NaCl. The addition of 300 micrograms potassium sorbate/ml to sucrose-supplemented media resulted in increased resistance to freezing in cultures previously incubated at 21 degrees C. Sensitivity to freezing increased when cultures were incubated at 21 degrees C in media not supplemented with solutes. Glucose and sucrose provided the best protection against inactivation by heating and freezing, regardless of the presence of potassium sorbate in growth media.

Colony formation by sublethally heat-injured Zygosaccharomyces rouxii as affected by solutes in the recovery medium and procedure for sterilizing medium.

Recovery and colony formation by healthy and sublethally heat-injured cells of Zygosaccharomyces rouxii as influenced by the procedure for sterilizing recovery media (YM agar [YMA], wort agar, cornmeal agar, and oatmeal agar) were investigated. Media were supplemented with various concentrations of glucose, sucrose, glycerol, or sorbitol and sterilized by autoclaving (110 degrees C, 15 min) and by repeated treatment with steam (100 degrees C). An increase in sensitivity was observed when heat-injured cells were plated on glucose-supplemented YMA at an aw of 0.880 compared with aws of 0.933 and 0.998. Colonies which developed from unheated and heated cells on YMA at aws of 0.998 and 0.933 generally exceeded 0.5 mm in diameter within 3.5 to 4 days of incubation at 25 degrees C, whereas colonies formed on YMA at an aw of 0.880 typically did not exceed 0.5 mm in diameter until after 5.5 to 6.5 days of incubation. The number of colonies exceeding 0.5 mm in diameter which were formed by heat-injured cells on YMA at an aw of 0.880 was 2 to 3 logs less than the total number of colonies detected, i.e., on YMA at an aw of 0.933 and using no limits of exclusion based on colony diameter. A substantial portion of cells which survived heat treatment were sublethally injured as evidenced by increased sensitivity to a suboptimum aw (0.880). In no instance was recovery of Z. rouxii significantly affected by medium sterilization procedure when glucose or sorbitol was used as the aw-suppressing solute.(ABSTRACT TRUNCATED AT 250 WORDS)

Efficacy of direct plating media for recovering Listeria monocytogenes from foods.

Studies were done to evaluate 14 direct plating media for their suitability to recover Listeria monocytogenes strain Scott A from pasteurized whole milk, chocolate ice cream mix, Brie cheese and raw cabbage. Healthy cells were inoculated into foods to achieve viable populations of 10(2), 10(4), or 10(6) cells/ml(g). Bind's Acriflavine Agar, Trypaflavine Nalidixic Acid Serum Agar, Listeria Transport Enrichment Agar, Doyle and Schoeni Selective Enrichment Agar (DSSEA) and Modified DSSEA were not suitable for recovering L. monocytogenes from milk and Brie cheese and were therefore not evaluated as direct plating media for recovering the organism from ice cream mix and cabbage. McBride Listeria Agar (MLA), Gum Base Nalidixic Acid Tryptone Soya Agar (GBNTSA), Modified Despierres Agar (MDA) and Modified MLA (MMLA) performed best for recovering all inoculum populations from milk and ice cream mix. Enumeration of L. monocytogenes on several test media was complicated by the growth of large numbers of background microflora present in cabbage and Brie cheese, especially at the lowest test inoculum (10(2)/g). Generally, complete recovery of L. monocytogenes from Brie cheese and cabbage was attained on media when the inoculum population was greater than or equal to 10(4) cells/g. For Brie cheese, MLA, MDA, MMLA and Dominguez Rodriguez Isolation Agar were superior for recovering L. monocytogenes, while GBNTSA, DLEA, MDA and MMLA were best for recovering the organism from cabbage. Results of this experiment indicate that direct-plating procedures, without prior enrichment, can successfully be utilized for recovering L. monocytogenes from foods such as pasteurized milk and ice cream mix which contain low populations of background microflora. However, recovery of L. monocytogenes from foods such as raw cabbage and Brie cheese, which contain high populations of other microorganisms, was not satisfactory using the direct-plating procedures evaluated in this investigation.

Influence of modified-atmosphere storage on the growth of uninjured and heat-injured Aeromonas hydrophila.

The growth of uninjured and heat-injured Aeromonas hydrophila incubated at 5 degrees C (22 days) and 30 degrees C (31 h) under air, N2, and CO2 was investigated. At 30 degrees C, the growth patterns of cells on brain heart infusion agar incubated under air and N2 were similar, although slight differences in the lengths of the lag phases and the final populations were detected. The lag phases of cells incubated under air and N2 were substantially longer at 5 degrees C than at 30 degrees C. The population of uninjured A. hydrophila incubated at 5 degrees C under air and N2 remained constant, whereas the number of injured cells declined before the exponential growth phase. Growth at 5 degrees C was enhanced when uninjured and heat-injured A. hydrophila were incubated under N2. At 30 degrees C, cells incubated under CO2 exhibited noticeably longer lag phases and lower growth rates than did cells incubated under air and N2. The viable populations of uninjured and heat-injured cells incubated at 5 degrees C under CO2 declined steadily throughout incubation.

Evaluation of selective direct plating media for their suitability to recover uninjured, heat-injured, and freeze-injured Listeria monocytogenes from foods.

Six direct plating media were evaluated for their suitability to recover uninjured, heat-injured, and freeze-injured cells of four strains of Listeria monocytogenes from four foods. Cells were inoculated into foods to achieve ca. 10(2) to 10(3), 10(4) to 10(5), or 10(5) to 10(6) viable cells per ml or g (low, medium, and high populations, respectively). No appreciable differences in recovery of the four test strains within a treatment were observed. Generally, recovery on all test media was similar and not markedly affected by freeze treatment. Modified Despierres agar and modified McBride Listeria agar yielded poorer recovery of heat-injured cells than did McBride Listeria agar and gum base-nalidixic acid-tryptone soya agar. Overall, gum base-nalidixic acid-tryptone soya agar was best for recovering L. monocytogenes from pasteurized milk and chocolate ice cream mix. Enumeration was complicated by the growth of background microflora present in Brie cheese and cabbage, especially at the low inoculum. Dominguez Rodriguez isolation agar was superior for recovering L. monocytogenes from Brie cheese, whereas modified Despierres agar was best for recovering the organism from cabbage. Direct plating procedures can successfully be utilized for recovering healthy and injured L. monocytogenes from foods containing low populations of background microflora.

Direct plating technique for enumeration of Listeria monocytogenes in foods.

The advantages and disadvantages of various techniques for detecting and enumerating Listeria monocytogenes in foods are reviewed, and results from a study designed to compare 14 direct plating media for their suitability to recover uninjured cells of L. monocytogenes from 4 foods are summarized. McBride Listeria agar (MLA), gum base nalidixic acid tryptone soy agar (GBNTSA), modified Despierres agar (MDA), and modified MLA (MMLA) performed best for recovering all inoculum populations from milk and ice cream mix. For Brie cheese, MLA, MDA, MMLA, and Dominguez Rodriguez isolation agar were superior for recovering L. monocytogenes; GBNTSA, MDA, MMLA, and Donnelly's Listeria enrichment agar were best for recovering the organism from cabbage. Direct plating procedures without prior enrichment can be utilized successfully for recovering L. monocytogenes from foods such as pasteurized milk and ice cream mix, which contain low populations of background microflora. However, recovery of L. monocytogenes from foods such as raw cabbage and Brie cheese, which contain high populations of other microorganisms, was not satisfactory using direct plating procedures.