Biomedical subjects

M A Drake

Publications and source records attributed to M A Drake.

At least 19 recordsLinked to original sources

PubMed2006

Validation of a green fluorescent protein-labeled strain of Vibrio vulnificus for use in the evaluation of postharvest strategies for handling of raw oysters.

In this paper we describe a biological indicator which can be used to study the behavior of Vibrio vulnificus, an important molluscan shellfish-associated human pathogen. A V. vulnificus ATCC 27562 derivative that expresses green fluorescent protein (GFP) and kanamycin resistance was constructed using conjugation. Strain validation was performed by comparing the GFP-expressing strain (Vv-GFP) and the wild-type strain (Vv-WT) with respect to growth characteristics, heat tolerance (45 degrees C), freeze-thaw tolerance (-20(o) and -80 degrees C), acid tolerance (pH 5.0, 4.0, and 3.5), cold storage tolerance (5 degrees C), cold adaptation (15 degrees C), and response to starvation. Levels of recovery were evaluated using nonselective medium (tryptic soy agar containing 2% NaCl) with and without sodium pyruvate. The indicator strain was subsequently used to evaluate the survival of V. vulnificus in oysters exposed to organic acids (citric and acetic acids) and various cooling regimens. In most cases, Vv-GFP was comparable to Vv-WT with respect to growth and survival upon exposure to various biological stressors; when differences between the GFP-expressing and parent strains occurred, they usually disappeared when sodium pyruvate was added to media. When V. vulnificus was inoculated into shellstock oysters, the counts dropped 2 log(10) after 11 to 12 days of refrigerated storage, regardless of the way in which the oysters were initially cooled. Steeper population declines after 12 days of refrigerated storage were observed for both iced and refrigerated products than for slowly cooled product and product held under conservative harvest conditions. By the end of the refrigeration storage study (22 days), the counts of Vv-GFP in iced and refrigerated oysters had reached the limit of detection (10(2) CFU/oyster), but slowly cooled oysters and oysters stored under conservative harvest conditions still contained approximately 10(3) and >10(4) CFU V. vulnificus/oyster by day 22, respectively. The Vv-GFP levels in the oyster meat remained stable for up to 24 h when the meat was exposed to acidic conditions at various pH values. Ease of detection and comparability to the wild-type parent make Vv-GFP a good candidate for use in studying the behavior of V. vulnificus upon exposure to sublethal stressors that might be encountered during postharvest handling of molluscan shellfish.

Animals↗

PubMed2006

Flavor profiles of full-fat and reduced-fat cheese and cheese fat made from aged Cheddar with the fat removed using a novel process.

Many consumers are concerned with fat intake. However, many reduced-fat foods, including reduced-fat cheese, lack robust flavors. The objectives of this study were to characterize the flavors found in full-fat cheese, cheese fat, and reduced-fat cheese made from aged Cheddar using a novel process to remove the fat (Nelson and Barbano, 2004). Two full-fat, aged cheeses (9 and 39 mo) were selected, and the fat was removed using the novel fat removal process. Full-fat cheeses, shredded and reformed full-fat cheeses, corresponding reduced-fat cheeses, and cheese fats were then analyzed using descriptive sensory and instrumental analysis followed by consumer acceptance testing. Cheeses were extracted with diethyl ether followed by isolation of volatile material by high vacuum distillation. Volatile extracts were analyzed using gas chromatography/ olfactometry with aroma extract dilution analysis. Selected compounds were quantified. The 39-mo cheese was characterized by fruity and sulfur notes, and the 9-mo-old cheese was characterized by a spicy/brothy flavor. Reduced-fat cheeses had similar flavor profiles with no difference in most sensory attributes to corresponding full-fat cheeses. Sensory profiles of the cheese fats were characterized by low intensities of the prominent flavors found in the full-fat cheeses. Instrumental analysis revealed similar trends. Consistent with sensory analysis, there were lower concentrations and log(3) flavor dilution factors for most compounds in the cheese fats compared with both the reduced- and full-fat cheeses, regardless of compound polarity. Consumers found the intensity of flavor in the reduced-fat cheese to be equal to the full-fat cheeses. This study demonstrated that when fat was removed from aged full-fat Cheddar cheese, most of the flavor and flavor compounds remained in the cheese and were not removed with the fat.

Cheese↗

PubMed2006

Enhanced nutty flavor formation in cheddar cheese made with a malty Lactococcus lactis adjunct culture.

Nutty flavor in Cheddar cheese is desirable, and recent research demonstrated that 2- and 3-methyl butanal and 2-methyl propanal were primary sources of nutty flavors in Cheddar. Because malty strains of Lac-tococcus lactis (formerly Streptococcus lactis var. malti-genes) are characterized by the efficient production of these and other Strecker aldehydes during growth, this study investigated the influence of a malty L. lactis adjunct culture on nutty flavor development in Cheddar cheese. Cheeses made with different adjunct levels (0, 10(4) cfu/mL, and 10(5) cfu/mL) were ripened at 5 or 13 degrees C and analyzed after 1 wk, 4 mo, and 8 mo by a combination of instrumental and sensory methods to characterize nutty flavor development. Cheeses ripened at 13 degrees C developed aged flavors (brothy, sulfur, and nutty flavors) more rapidly than cheeses held at 5 degrees C. Additionally, cheeses made with the adjunct culture showed more rapid and more intense nutty flavor development than control cheeses. Cheeses that had higher intensities of nutty flavors also had a higher concentration of 2/3-methyl butanal and 2-methyl propanal compared with control cheeses, which again confirmed that these compounds are a source of nutty flavor in Cheddar cheese. Results from this study provide a simple methodology for cheese manufacturers to obtain consistent nutty flavor in Cheddar cheese.

Aldehydes↗

PubMed2006

Mexican chihuahua cheese: sensory profiles of young cheese.

Sensory profiles of fresh semihard Chihuahua cheese produced in the northern Mexican state of Chihuahua were developed to characterize the flavors and textures of this traditionally made Hispanic-style cheese. Multiple allotments of Chihuahua cheese, 9 brands made with raw milk (RM) and 5 brands made with pasteurized milk (PM), were obtained within 3 d of manufacture from 12 different cheese plants throughout Chihuahua, México. Cheeses were shipped overnight to Wyndmoor, Pennsylvania, and flavor analyses were conducted within 14 to 18 d after manufacture. Four brands (2 RM and 2 PM cheeses) were then selected and multiple allotments were shipped at 3 distinct seasons over a 1-yr period for evaluation of flavor and texture. Microbial analysis was conducted prior to testing to ensure product safety. Descriptive analyses of cheese flavors and textures were conducted with panelists trained to use a universal or product-specific Spectrum intensity scale, respectively. Sensory profiles of cheeses varied among the different manufacturers. The most prominent flavor attributes were salty, sour, diacetyl, cooked, whey, bitter, and milk-fat. The RM cheeses had more intense sour, bitter, and prickle scores than the PM cheeses. Many cheese texture attributes were similar, but RM cheeses were perceived as softer than PM cheeses. As the demand for Hispanic-style cheeses increases, defining and understanding the sensory attributes of traditionally made Mexican cheeses provides guidance to cheese manufacturers as new ways are explored to improve the production and shelf life of the cheeses.

Animals↗

PubMed2005

Characterization of dried whey protein concentrate and isolate flavor.

The flavor of whey protein concentrates (WPC 80) and whey protein isolates (WPI) was studied using instrumental and sensory techniques. Four WPC 80 and 4 WPI, less than 3 mo old, were collected in duplicate from 6 manufacturers in the United States. Samples were rehydrated and evaluated in duplicate by descriptive sensory analysis. Duplicate samples with internal standards were extracted with diethyl ether. Extracts were then distilled to remove nonvolatile material using high vacuum distillation. Volatile extracts were analyzed using gas chromatography/olfactometry with post peak intensity analysis and aroma extract dilution analysis. Compounds were identified by comparison of retention indices, odor properties, and gas chromatography/mass spectrometry against reference standards. Whey proteins exhibited sweet aromatic, cardboard/wet paper, animal/wet dog, soapy, brothy, cucumber, and cooked/milky flavors, along with the basic taste bitter, and the feeling factor astringency. Key volatile flavor compounds in WPC 80 and WPI were butanoic acid (cheesy), 2-acetyl-1-pyrroline (popcorn), 2-methyl-3-furanthiol (brothy/burnt), 2,5-dimethyl-4-hydroxy-3-(2H)-furanone (maple/spicy), 2-nonenal (fatty/old books), (E,Z)-2,6-nonadienal (cucumber), and (E,Z)-2,4-decadienal (fatty/oxidized). This baseline data on flavor and flavor sources in whey proteins will aid ongoing and future research and will help to identify the most appropriate whey ingredients to use to control or minimize flavor variability in whey enhanced products.

Alcohols↗

PubMed2005

Comparison of sensory, microbiological, and biochemical parameters of microwave versus indirect UHT fluid skim milk during storage.

Shelf-stable milk could benefit from sensory quality improvement. Current methods of heating cause flavor and nutrient degradation through exposure to overheated thermal exchange surfaces. Rapid heating with microwaves followed by sudden cooling could reduce or eliminate this problem. The objectives for this study were focused on designing and implementing continuous microwave thermal processing of skim fluid milks (white and chocolate) to compare sensory, microbiological, and biochemical parameters with conventionally prepared, indirect UHT milks. All test products were aseptically packaged and stored at ambient temperature for 12 mo. Every 3 mo, samples were taken for microbiological testing, reactive sulfhydryl determinations, active enzyme analysis, instrumental viscosity readings, color measurements, and descriptive sensory evaluation. Microbiological plate counts were negative on all milks at each time point. Enzymatic assays showed that plasmin was inactivated by both heat treatments. 5,5'-dithio-bis(2-nitrobenzoic acid) analysis, a measure of reactive sulfhydryl (-SH-) groups, showed that the initial thiol content was not significantly different between the microwave-processed and UHT-treated milks. However, both heating methods resulted in an increased thiol level compared with conventionally pasteurized milk samples due to the higher temperatures attained. Sulfhydryl oxidase, a milk enzyme that catalyzes disulfide bond formation using a variety of protein substrates, retained activity following microwave processing, and decreased during storage. Viscosity values were essentially equivalent in microwave- and UHT-heated white skim milks. Sensory analyses established that UHT-treated milks were visibly darker, and exhibited higher caramelized and stale/fatty flavors with increased astringency compared with the microwave samples. Sweet aromatic flavor and sweet taste decreased during storage in both UHT and microwave milk products, whereas stale/fatty flavors increased over time. Sensory effects were more apparent in white milks than in chocolate varieties. These studies suggest that microwave technology may provide a useful alternative processing method for delivery of aseptic milk products that retain a long shelf life.

Animals↗

PubMed2004

Characterization of nutty flavor in cheddar cheese.

The objectives of this study were to determine the volatile components responsible for the sensory perception of nutty flavor in Cheddar cheese. Cheddar cheeses with and without nutty flavors were selected by descriptive sensory analysis. Volatile aroma components from Cheddar cheeses with and without nutty flavors were isolated and characterized using solvent extraction with high vacuum distillation, dynamic headspace analysis, gas chromatography-olfactometry, and gas chromatography-mass spectrometry. More than 50 aroma-active compounds were detected in Cheddar cheeses. Consistent differences were observed between nutty and not nutty Cheddar cheeses. Strecker aldehydes were detected in higher amounts in Cheddar cheeses with nutty flavors compared with Cheddar cheeses without nutty flavors. Strecker aldehydes, dimethyl sulfide, and propionic acid were evaluated in young and aged Cheddar cheese models for nutty flavor by descriptive sensory analysis. Dimethyl sulfide and propionic acid did not contribute to nutty flavor in Cheddar cheese. The addition of Strecker aldehydes to young (<4 mo old) Cheddar cheese models resulted in nutty/malty flavor perceived by sensory analysis. When Strecker aldehydes were incorporated into aged (>9 mo old) Cheddar cheese models, nutty flavor perception increased. Strecker aldehydes contribute to nutty flavor in aged Cheddar cheese.

Aldehydes↗

PubMed2004

Influence of adjunct cultures on volatile free fatty acids in reduced-fat Edam cheeses.

The effects of the adjunct cultures Lactococcus lactis ssp. diacetylactis, Brevibacterium linens BL2, Lactobacillus helveticus LH212, and Lactobacillus reuteri ATCC 23272 on volatile free fatty acid production in reduced-fat Edam cheese were studied. Lipase activity evaluation using p-nitrophenyl fatty acid ester substrates indicated that L. lactis ssp. diacetylactis showed the highest activity among the 4 adjunct cultures. Full-fat and 33% reduced-fat control cheeses (no adjunct) were made along with 5 treatments of reduced-fat cheeses, which included individual, and a mixture of the adjunct cultures. Volatile free fatty acids of cheeses were analyzed using static headspace analysis with 4-bromofluorobenzene as an internal standard. Changes in volatile free fatty acid concentrations were found in headspace gas of cheeses after 3-and 6-mo ripening. Acetic acid was the most abundant acid detected throughout ripening. Full-fat cheese had the highest relative amount of propionic acid among the cheeses. Certain adjunct cultures had a definite role in lipolysis at particular times. Reduced-fat cheese with L. lactis ssp. diacetylactis at 3-mo showed the highest levels of butyric, isovaleric, n-valeric, iso-caproic, and n-caproic acid. Reduced-fat cheese with Lactobacillus reuteri at 6 mo produced the highest relative concentration of isocaproic, n-caproic, and heptanoic, and the highest relative concentration of total acids.

Acetic Acid↗

PubMed2004

Impact of cold and cold-acid stress on poststress tolerance and virulence factor expression of Escherichia coli O157:H7.

The effect of extended cold or cold-acid storage of Escherichia coli O157:H7 on subsequent acid tolerance, freeze-thaw survival, heat tolerance, and virulence factor (Shiga toxin, intimin, and hemolysin) expression was determined. Three E. coli O157:H7 strains were stressed at 4 degrees C in TSB or pH 5.5 TSB for 4 weeks. The acid (TSB [pH 2.0] or simulated gastric fluid [pH 1.5]) tolerance, freeze-thaw (-20 degrees C to 21 degrees C) survival, and heat (56 degrees C) tolerance of stressed cells were compared with those of control cells. The beta-galactosidase activities of stressed and control cells containing a lacZ gene fusion in the stx2, eaeA, or hlyA gene were determined following stress in TSB or pH 5.5 TSB at 37 degrees C and in the exponential and stationary phases. Cold and cold-acid stresses decreased acid tolerance (P < 0.05), with a larger decrease in acid tolerance being observed after cold stress than after cold-acid stress (P < 0.05). Cold stress increased freeze-thaw survival for all three strains (P < 0.05). Prior cold or cold-acid stress had no effect on virulence factor production (P > 0.05), although growth in acidic media (pH 5.5) enhanced eaeA and hlyA expression (P < 0.05). These results indicate that the prolonged storage of E. coli O157:H7 at 4 degrees C has substantial effects on freeze-thaw tolerance but does not affect subsequent virulence gene expression.

Adaptation, Physiological↗

PubMed2003

The impact of fortification with conjugated linoleic acid (CLA) on the quality of fluid milk.

The effects of added conjugated linoleic acid (CLA) on the sensory, chemical, and physical characteristics of 2% total fat (wt/wt) fluid milk were studied. Milks with 2% (wt/wt) total fat (2% CLA, 1% CLA 1% milkfat, 2% milkfat) were made by the addition of cream or CLA triglyceride oil into skim milk followed by HTST pasteurization and homogenization. The effects of adding vitamin E (200 ppm) and rosemary extract (0.1% wt/wt based on fat content) were investigated to prevent lipid oxidation. HTST pasteurization resulted in a significant decrease of the cis-9/trans-11 isomer and other minor CLA isomers. The cis-9/trans-11 isomer concentration remained stable through 2 wk of refrigerated storage. A significant loss of both the cis-9/trans-11 and the cis-10/trans-12 isomers occurred after 3 wk of refrigerated storage. The loss was attributed to lipase activity from excessive microbial growth. No differences were found in hexanal or other common indicators of lipid oxidation between milks with or without added CLA (P > 0.05). Descriptive sensory analysis revealed that milks with 1 or 2% CLA exhibited low intensities of a "grassy/vegetable oil" flavor, not present in control milks. The antioxidant treatments were deemed to be ineffective, under the storage conditions of this study, and did not produce significant differences from the control samples (P > 0.05). CLA-Fortified milk had significantly lower L* and b* values compared with 2% milkfat milk. No significant differences existed in viscosity. Consumer acceptability scores (n = 100) were lower (P < 0.05) for CLA-fortified milks compared to control milks, but the addition of chocolate flavor increased acceptability (P < 0.05).

Adolescent↗

PubMed2003

Determining flavor and flavor variability in commercially produced liquid cheddar whey.

Dried whey and whey protein are important food ingredients. Functionality of whey products has been studied extensively. Flavor inconsistency and flavors which may carry through to the finished product can limit whey ingredient applications in dairy and nondairy foods. The goal of this research was to determine the flavor and flavor variability of commercially produced liquid Cheddar cheese whey. Liquid Cheddar cheese whey from five culture blends from two different stirred-curd Cheddar cheese manufacturing facilities was collected. Whey flavor was characterized using instrumental and sensory methods. Wide variation in whey headspace volatiles was observed between different manufacturing facilities (P < 0.05). Hexanal and diacetyl were two key volatiles that varied widely (P < 0.05). FFA profiles determined by solid-phase microextraction and degree of proteolysis of the whey samples were also different (P < 0.05). Differences in whey flavor profiles were also confirmed by descriptive sensory analysis (P < 0.05). Differences in liquid whey flavor were attributed to differences in milk source, processing and handling and starter culture blend. The flavor of liquid Cheddar cheese whey is variable and impacted by milk source and starter culture rotation. Results from this study will aid future studies that address the impact of liquid whey flavor variability on flavor of dried whey ingredients.

Amino Acids↗

PubMed2003

Effects of standardization of whole milk with dry milk protein concentrate on the yield and ripening of reduced-fat cheddar cheese.

Commercial milk protein concentrate (MPC) was used to standardize whole milk for reduced-fat Cheddar cheesemaking. Four replicate cheesemaking trials of three treatments (control, MPC1, and MPC2) were conducted. The control cheese (CC) was made from standardized milk (casein-to-fat ratio, C/F approximately 1.7) obtained by mixing skim milk and whole milk (WM); MPC1 and MPC2 cheeses were made from standardized milk (C/F approximately 1.8) obtained from mixing WM and MPC, except that commercial mesophilic starter was added at the rate of 1% to the CC and MPC1 and 2% to MPC2 vats. The addition of MPC doubled cheese yields and had insignificant effects on fat recoveries (approximately 94% in MPC1 and MPC2 vs. approximately 92% in CC) but increased significantly total solids recoveries (approximately 63% in CC vs. 63% in MPC1 and MPC2). Although minor differences were noted in the gross composition of the cheeses, both MPC1 and MPC2 cheeses had lower lactose contents (0.25 or 0.32%, respectively) than in CC (0.60%) 7 d post manufacture. Cheeses from all three treatments had approximately 10(9) cfu/g initial starter bacteria count. The nonstarter lactic acid bacteria (NSLAB) grew slowly in MPC1 and MPC2 cheeses during ripening compared to CC, and at the end of 6 mo of ripening, numbers of NSLAB in the CC were 1 to 2 log cycles higher than in MPC1 and MPC2 cheeses. Primary proteolysis, as noted by water-soluble N contents, was markedly slower in MPC1 and MPC2 cheeses compared to CC. The concentrations of total free amino acids were in decreasing order CC > MPC2 > MPC1 cheeses, suggesting slower secondary proteolysis in the MPC cheeses than in CC. Sensory analysis showed that MPC cheeses had lower brothy and bitter scores than CC. Increasing the amount of starter bacteria improved maturity in MPC cheese.

Animals↗

PubMed2003

The effect of application of cold natural smoke on the ripening of Cheddar cheese.

The present study was undertaken to study the effects of application of natural wood smoke on ripening of Cheddar cheese, and to determine the effects of smoking before or after ripening on cheese quality. A 20-kg block of Cheddar cheese obtained immediately after pressing was divided into six approximately 3-kg blocks and ripened at 8 degrees C for up to 270 d. One 3-kg block was taken after 1 d, 1, 3, 6, or 9 mo and smoked for 20 min, then returned to the ripening room for further ripening. Cheeses were sampled at intervals for lactobacilli counts, moisture, pH, and proteolysis. Sensory analysis was conducted on 6 and 9-mo-old cheeses by a trained sensory panel (n = 7). Results show that application of natural wood smoke did not significantly affect cheese pH or primary proteolysis during ripening. However, secondary proteolysis as assessed by the concentrations of free amino acids was generally higher in smoked cheeses than in control cheeses after 6 mo of ripening. Cheese smoked after 6 mo of ripening had better smoked flavor than that smoked after 9 mo of ripening. Cheese smoked after 3 mo of age and further ripened for 6 mo had the highest smoked flavor intensity. It is concluded that it is best to smoke cheese after ripening for at least 3 mo.

Cheese↗

PubMed2003

Relationships among rheological and sensorial properties of young cheeses.

This study investigated the sensory and rheological properties of young cheeses in order to better understand perceived cheese texture. Mozzarella and Monterey Jacks were tested at 4, 10, 17, and 38 d of age; process cheese was tested at 4 d. Rheological methods were used to determine the linear and nonlinear viscoelastic and fracture properties. A trained sensory panel developed a descriptive language and reference scales to evaluate cheese texture. All methods differentiated the cheeses by variety. Principal component analysis of sensory texture revealed that three principal components explained 96.1% of the total variation in the cheeses. The perception of firmness decreased as the cheeses aged, whereas the perception of springiness increased. Principal component analysis of the rheological parameters (three principal components: 87.9% of the variance) showed that the cheeses' solid-like response (storage modulus and fracture modulus) decreased during aging, while phase angle, maximum compliance, and retardation time increased. Analysis of the instrumental and sensory parameters (three principal components: 82.1% of the variance) revealed groupings of parameters according to cheese rigidity, resiliency, and chewdown texture. Rheological properties were highly associated with rigidity and resiliency, but less so with chewdown texture.

Cheese↗

PubMed2003

Acid adaptation and starvation effects on Shiga toxin production by Escherichia coli O157:H7.

Reverse transcription-polymerase chain reaction, enzyme-linked immunosorbent assay (ELISA), and gene fusion were used to determine differences in the expression of stx-II mRNA and the production of mature Stx protein following acid adaptation or starvation of Escherichia coli O157:H7 (ATCC 43895) and an isogenic rpoS mutant (FRIK 816-3) under static conditions and with shaking. The expression of stx-II mRNA in acid-adapted and starved cells was more extensive than that in nonstressed control cells. This effect was more pronounced for the rpoS mutant. Oxygenation (incubation with shaking) increased stx-II mRNA expression for both strains relative to the level of expression obtained with static conditions. ELISA results indicated that Stx production was enhanced more in the rpoS mutant than in its wild-type parent strain and that oxygenation enhanced Stx production for both strains but there were no detectable differences between stressed and nonstressed cells of either strain. The monitoring of the gene product of Stx-II alone with the use of stx-IIAB::lacZ gene fusions confirmed the induction of aeration and the absence of a stress effect for both the wild type and the rpoS mutant. These results indicate that oxygen enhances stx-II mRNA expression and Stx production in E. coli O157:H7. Stress conditions such as acid adaptation and starvation enhance stx-II toxin mRNA levels but do not enhance subsequent Stx toxin production.

Adaptation, Physiological↗

PubMed2003

Detection of viable Shiga toxin-producing Escherichia coli by quantitative competitive polymerase chain reaction.

With the use of Escherichia coli O157:H7 as a model, a procedure for the quantitative detection of viable Shiga toxin-producing E. coli (STEC) in broth and cooked ground beef enrichments with multiple-time point quantitative competitive polymerase chain reaction (QC-PCR) was developed. The A subunit (a 401-bp fragment) of the stx2 gene was chosen as a target sequence. Immunomagnetic separation (IMS) was used to isolate and concentrate cells from ground beef enrichments. Cell viability was confirmed on the basis of the quantitative increase in the signal of target bands from QC-PCR across multiple time points. The application of IMS increased detection limits relative to those for QC-PCR without IMS. E. coli O157:H7 inoculated at 0.20 CFU/g of cooked ground beef (25 g of ground beef plus 225 ml of Bacto modified EC medium plus novobiocin) was detected and confirmed to be viable in <15 h. A DNA-based molecular approach can be used to determine cell viability.

Colony Count, Microbial↗

PubMed2002

Optimization of reverse transcriptase PCR to detect viable Shiga-toxin-producing Escherichia coli.

The ability of reverse transcriptase PCR (RT-PCR) to detect viable Shiga-toxin-producing Escherichia coli (STEC) was investigated. Four primer sets, each targeting a specific region in the slt-II operon, were evaluated for their stringency and specificity for slt-II mRNA. STEC were evaluated for toxin expression under various conditions, including cell growth phase, growth medium, incubation temperature, and aeration. Following primer optimization, STEC were inoculated into Trypticase soy broth and cooked ground beef enrichments. Cells were harvested and RNA or DNA was extracted at 4, 8, 12, and 24 h. RT-PCR or PCR was conducted, and the products were visualized by gel electrophoresis and by Southern blots. mRNA targets were detected in 12-h cooked ground meat enrichments with an initial inoculum of 1 CFU/g. These results indicate that RT-PCR of E. coli slt-II mRNA is useful for detection of viable STEC in ground beef.

Animals↗

PubMed2002

Resistance of cold- and starvation-stressed Vibrio vulnificus to heat and freeze-thaw exposure.

The effects of cold storage and starvation on the subsequent heat resistance and freeze-thaw resistance of Vibrio vulnificus were studied. Three strains of V. vulnificus were evaluated. Cold stress had no effect on freeze-thaw resistance (P > 0.05). Starvation enhanced freeze-thaw resistance for one strain compared to controls (P < 0.05). V. vulnificus was not heat resistant; control populations were inactivated within 12 min at 47 degrees C. Starvation increased heat tolerance for one strain, but differences were small from a processing perspective (P < 0.05). Cold stress had no effect on heat resistance (P > 0.05). Cold adaptation (holding 4 h at 15 degrees C) enhanced cold temperature (5 degrees C) tolerance. This information will be helpful in the development of methods to minimize V. vulnificus risk.

Adaptation, Physiological↗