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Biomedical subjects

M P Doyle

Publications and source records attributed to M P Doyle.

At least 19 recordsLinked to original sources
PubMed2006

Reducing the carriage of foodborne pathogens in livestock and poultry.

Several foodborne pathogens, including Salmonella species and campylobacters, are common contaminants in poultry and livestock. Typically, these pathogens are carried in the animal's intestinal tract asymptomatically; however, they can be shed in feces in large populations and be transmitted by other vectors from feces to animals, produce, or humans. A wide array of interventions has been developed to reduce the carriage of foodborne pathogens in poultry and livestock, including genetic selection of animals resistant to colonization, treatments to prevent vertical transmission of enteric pathogens, sanitation practices to prevent contamination on the farm and during transportation, elimination of pathogens from feed and water, feed and water additives that create an adverse environment for colonization by the pathogen, and biological treatments that directly or indirectly inactivate the pathogen within the host. To successfully reduce the carriage of foodborne pathogens, it is likely that a combination of intervention strategies will be required.

Animal Feed↗
PubMed2006

Role of colanic acid exopolysaccharide in the survival of enterohaemorrhagic Escherichia coli O157:H7 in simulated gastrointestinal fluids.

AIM: This study evaluated the production of colanic acid (CA) exopolysaccharide (EPS) by Escherichia coli O157:H7 in relation to the pathogen's ability to survive under acidic conditions simulating the environment in the human gastrointestinal tract. METHODS AND RESULTS: Escherichia coli O157:H7 W6-13 and its CA-deficient mutant M4020 were examined for their resistance to bile salts, and their ability to survive in simulated gastric fluid containing pepsin (pH 2.0) and simulated intestinal fluid containing pancreatin (pH 8.0). The effect of acid adaptation at pH 5.5 on the survival of E. coli O157:H7 in simulated gastric fluid was also determined. The results indicated that the survivability of M4020, under conditions simulating the environment in the human gastrointestinal tract, reduced more drastically than the viability of W6-13. The presence of bile salts had a slight effect on both types of E. coli O157:H7 cells. The loss of CA did not change the ability of M4020 to respond to acid adaptation. CONCLUSION: The EPS CA may serve as a protective barrier to E. coli O157:H7 for its survival in the human gastrointestinal tract. SIGNIFICANCE AND IMPACT OF THE STUDY: The study contributes to a better understanding of the EPS affecting the ability of E. coli O157:H7 to combat acid stress.

Escherichia coli O157↗
PubMed2006

Effect of fat content on infection by Listeria monocytogenes in a mouse model.

An estimated 2,500 cases of listeriosis occur annually in the United States. Listeriosis is particularly severe among pregnant women and immunocompromised individuals. Little is known regarding the effect of the food matrix on the ability of L. monocytogenes to survive in the gastrointestinal tract and cause systemic infection. Mice were inoculated with various doses of L. monocytogenes in skim milk, Half & Half, or whipping cream to determine whether differences in milk fat content influence the ability of L. monocytogenes to survive passage through the gut and infect the liver or spleen. The number of fecal samples positive for L. monocytogenes increased with increasing doses of L. monocytogenes for all three vehicles. The number of L. monocytogenes cells isolated from liver or spleen of mice dosed with L. monocytogenes was not significantly different among treatment vehicles. Dose-response models revealed that as the dosage of L. monocytogenes was increased in different milk vehicles, the number of L. monocytogenes cells in liver or spleen also increased. Although fat content of food had no dose-dependent effect on L. monocytogenes infection in the murine gastrointestinal tract, we cannot discount the possibility that it may be a factor in L. monocytogenes infections of humans because of differences in the physiology of gastrointestinal tracts of mice and humans.

Animals↗
PubMed2004

Ethanol-mediated variations in cellular fatty acid composition and protein profiles of two genotypically different strains of Escherichia coli O157:H7.

Two strains of Escherichia coli O157:H7 were grown in tryptic soy broth (TSB, pH 7.1) supplemented with 0, 2.5, 5.0, 7.5, and 10% ethanol at 30 degrees C for up to 54 h. Growth rates in TSB supplemented with 0, 2.5, and 5.0% ethanol decreased with an increase in ethanol concentration. Growth was not observed in TSB supplemented with 7.5 or 10% ethanol. The pH of TSB containing 5.0% ethanol decreased to 5.8 within 12 h and then increased to 7.0 at 54 h. The ethanol content in TSB supplemented with 2.5 or 5.0% ethanol did not change substantially during the first 36 h of incubation but decreased slightly thereafter, indicating utilization or degradation of ethanol by both strains. Glucose was depleted in TSB supplemented with 0, 2.5, or 5.0% ethanol within 12 h. Cells grown under ethanol stress contained a higher amount of fatty acids. With the exceptions of cis-oleic acid and nonadecanoic acid, larger amounts of fatty acid were present in stationary-phase cells of the two strains grown in TSB supplemented with 5.0% ethanol for 30 h than in cells grown in TSB without ethanol for 22 h. The trans-oleic acid content was 10-fold higher in the cells grown in TSB with 5.0% ethanol than those grown in TSB without ethanol. In contrast, cis-oleic acid was not detected in ethanol-stressed cells but was present at concentrations of 0.32 and 0.36 mg/g of cells of the two strains grown in TSB without ethanol. Protein content was higher in ethanol-stressed cells than in nonstressed cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profiles varied qualitatively as affected by the strain and the presence of ethanol in TSB. An ethanol-mediated protein (28 kDa) was observed in the ethanol-stressed cells but not in control cells. It is concluded that the two test strains of E. coli O157:H7 underwent phenotypic modifications in cellular fatty acid composition and protein profiles in response to ethanol stress. The potential for cross protection against subsequent stresses applied in food preservation technologies as a result of these changes is under investigation.

DNA, Bacterial↗
PubMed2002

Consumer acceptance of raw apples treated with an antibacterial solution designed for home use.

An antibacterial treatment consisting of 1.5% lactic acid plus 1.5% hydrogen peroxide at 40 degrees C for 15 min was effective in reducing foodborne bacterial pathogens on raw apples. However, the effects of this treatment on an apple's sensory characteristics and the extent of consumers' willingness to use the treatment at home were not known. This study was undertaken to determine the sensory acceptability and chemical characteristics (pH, soluble solids, and total acidity) of apples subjected to the sanitizing treatment and to obtain information on consumers' purchase behavior, apple handling and consumption practices, and willingness to use an antibacterial treatment. Untrained consumers (n = 80) evaluated the appearance, color, aroma, flavor, texture, and overall appeal of untreated (control) and treated Red Delicious apples that had been stored at 5 degrees C for 0, 6, and 10 days. Panelists used a nine-point hedonic scale (1 = "dislike extremely"; 5 = "neither like nor dislike"; 9 = "like extremely") to evaluate sensory acceptability. Treatment and storage had no significant effect on the appearance, color, or aroma of the samples. Flavor ratings ranged from 6.2 ("like slightly") to 7.0 ("like moderately"). There was no significant difference among any of the control and treated apples stored for 0 days or among those stored for 6 days. Although apples stored for 10 days received the lowest ratings (6.2 to 6.3), they still had an acceptable flavor (6, "like slightly"), and panelists could not perceive differences between the control samples and the treated samples on day 10. The same trends were noted in texture ratings and in overall liking ratings. Treatment and storage had a minimal effect on pH (range, 3.96 to 4.02), soluble solids (range, 11.8 to 12.9 degrees Brix), and total acidity (range, 0.20 to 0.23% malic acid), which are important for apple flavor. Many consumers (87%) were concerned about fruit safety, and 53.2% were willing to try an antibacterial treatment at home. However, 74% would not be willing to use it if a 15-min heating-and-soaking step were required. Implementation of the treatment may be more feasible in the packinghouse than in the home.

Consumer Behavior↗
PubMed2002

Fate of Escherichia coli O157:H7 in coleslaw during storage.

An outbreak of Escherichia coli O157:H7 infection associated with the consumption of coleslaw in several units of a restaurant chain prompted a study to determine the fate of the pathogen in two commercial coleslaw preparations (pH 4.3 and 4.5) held at 4, 11, and 21 degrees C for 3 days. At an initial population of 5.3 log10 CFU/g of coleslaw, E. coli O157:H7 did not grow in either coleslaw stored at the three temperatures. Rather, the population of E. coli O157:H7 decreased by 0.1 to 0.5 log10 CFU/g within 3 days. The greatest reduction (0.4 and 0.5 log10 CFU/g) in population occurred at 21 degrees C, whereas only slight decreases (0.1 to 0.2 log10 CFU/g) occurred at 4 and 11 degrees C. A pH of 4.3 to 4.5 of coleslaw had little effect on reducing E. coli O157:H7 populations. Results suggest that the tolerance of E. coli O157:H7 to acid pH, not temperature abuse, is a major factor influencing the pathogen's fate in restaurant-prepared coleslaw.

Colony Count, Microbial↗
PubMed2002

Consumer acceptance of fresh-cut iceberg lettuce treated with 2% hydrogen peroxide and mild heat.

An antibacterial treatment consisting of 2% hydrogen peroxide at 50 degrees C for 60 s was evaluated for its effects on the sensory quality of fresh-cut iceberg lettuce. Treated lettuce was packaged in polyethylene film by a protocol used in the fresh-cut produce industry and then stored along with untreated controls for 3, 10, and 15 days at 5 degrees C. Gas chromatographic analysis confirmed that the desired initial volume (approximately 6 liters) and oxygen gas content (approximately 10%) were maintained during storage. Consumers from the local community who were the primary shoppers for their households and who purchased and ate lettuce regularly evaluated the appearance, color, aroma, flavor, and texture of the lettuce and their overall liking of the lettuce. Forty consumers were recruited to evaluate processing replication 1 on 26 April, and another 40 were chosen to evaluate processing replication 2 on 3 May. Because replication differences were observed in instrumental color measurements and in mean sensory ratings, replications were not pooled for the two test dates. Lettuce purchased for processing replication 2 was considerably greener than that used in replication 1. Overall, the antibacterial treatment was more effective than the control treatment in maintaining sensory quality over 15 days of storage, provided that the lettuce was initially intensely green. Three-fourths of the participants indicated that they would be willing to buy precut packaged lettuce that had already been treated at the packinghouse or processing plant with an antibacterial solution, and of these participants, 62.5% indicated that they would be willing to pay 5 to 10 cents more per bag.

Anti-Infective Agents, Local↗
PubMed2001

Factors influencing the detection and enumeration of Escherichia coli O157:H7 on alfalfa seeds.

Isolating Escherichia coli O157:H7 from batches of alfalfa seeds used to produce sprouts implicated in human illness has been difficult, perhaps due to nonhomogenous and very low-level contamination and inaccessibility of the pathogen entrapped in protected areas of the seed coat. We evaluated the effectiveness of various treatments in releasing E. coli O157:H7 from seeds. The influence of homogenization (blending or stomaching for 1 or 2 min), rinsing method (shaking for 5 min), soaking time (0. 1, 3, 6, or 15 h), soaking temperature (4 or 21 degrees C), and the addition of surfactants (0.1%, 0.5%, or 1.0% Tween 80 or Span 20) to rinse water was determined. Blending or stomaching for 1 or 2 min, and soaking for 1 h or longer at 4 or 21 degrees C, respectively, resulted in maximum release of E. coli O157:H7 from seeds. Soaking seeds at 37 degrees C for 15 h increased cell populations of E. coli O157:H7 by approximately 3.6 log10 CFU/g, likely due to bacterial growth. The maximum number of cells released from seeds by rinse water containing 1.0% Span 20 was at 21 degrees C, whereas at 37 degrees C, 0.1% or 0.5% Tween 80 was more effective. Detection of E. coli O157:H7 on seeds stored at 37 degrees C for up to 13 weeks and on sprouts derived from these seeds was compared. E. coli O157:H7 inoculated on seeds at 2.0 log10 CFU/g was detected after storage of seeds for up to 8 weeks at 37 degrees C and in sprouts produced from the seeds. The pathogen was not detected on seeds stored for 13 weeks at 37 degrees C and was not isolated from sprouts produced from these seeds. Identifying seed treatment methods that enhance removal of E. coli O157:H7 from alfalfa seeds can aid the isolation and enumeration of the pathogen on seeds. With a combination of optimal conditions for detecting the pathogen, i.e. soaking seeds for 1 h and pummeling seeds for 1 min, an enrichment step in modified tryptic soy broth (TSB), and the use of immunomagnetic beads for separation of E. coli O157:H7 cells, E. coli O157:H7 was detected in alfalfa seeds incubated at 37 degrees C for up to 8 weeks as effectively as in sprouts produced from the seeds.

Colony Count, Microbial↗
PubMed2001

High selectivity from configurational match/mismatch in carbon-hydrogen insertion reactions of steroidal diazoacetates catalyzed by chiral dirhodium(II) carboxamidates.

Diazo decomposition of steroidal diazoacetates, where the point of attachment is the 3-position of the steroid A-ring, catalyzed by chiral dirhodium(II) carboxamidates results in products from carbon-hydrogen insertion in high yield and selectivities. Use of S-configured catalysts shows a distinctive preference for insertion into the 3-position to form beta-lactone products. The R-configured catalysts direct insertion preferentially to the equatorial C-H bond at the 2-position. Substituents or functional groups at the 5/6-position prevent C-H insertion from taking place at the 4-position. Even in the best case with the 5/6-positions fully saturated, however, insertion into the 3-position remains competitive with insertion into the 4-position. Corresponding 3-substituted phenyldiazoacetates give only beta-lactone products, and selectivity here is highest with chiral dirhodium(II) prolinate catalysts. A model is presented to explain these results. Overall, this methodology is versatile for functionalization of the steroid A-ring at positions 2 and 3.

Azo Compounds↗
PubMed2001

Experimental and field studies of Escherichia coli O157:H7 in white-tailed deer.

Studies were conducted to evaluate fecal shedding of Escherichia coli O157:H7 in a small group of inoculated deer, determine the prevalence of the bacterium in free-ranging white-tailed deer, and elucidate relationships between E. coli O157:H7 in wild deer and domestic cattle at the same site. Six young, white-tailed deer were orally administered 10(8) CFU of E. coli O157:H7. Inoculated deer were shedding E. coli O157:H7 by 1 day postinoculation (DPI) and continued to shed decreasing numbers of the bacteria throughout the 26-day trial. Horizontal transmission to an uninoculated deer was demonstrated. Although E. coli O157:H7 bacteria were recovered from the gastrointestinal tracts of deer necropsied from 4 to 26 DPI, attaching and effacing lesions were not apparent in any deer. Results are similar to those of inoculation studies in calves and sheep. In field studies, E. coli O157 was not detected in 310 fresh deer fecal samples collected from the ground. It was detected in feces, but not in meat, from 3 of 469 free-ranging deer in 1997. In 1998, E. coli O157 was not detected in 140 deer at the single positive site found in 1997; however, it was recovered from 13 of 305 dairy and beef cattle at the same location. Isolates of E. coli O157:H7 from deer and cattle at this site differed with respect to pulsed-field gel electrophoresis patterns and genes encoding Shiga toxins. The low overall prevalence of E. coli O157:H7 and the identification of only one site with positive deer suggest that wild deer are not a major reservoir of E. coli O157:H7 in the southeastern United States. However, there may be individual locations where deer sporadically harbor the bacterium, and venison should be handled with the same precautions recommended for beef, pork, and poultry.

Animals↗
PubMed2001

Insertion mutagenesis of wca reduces acid and heat tolerance of enterohemorrhagic Escherichia coli O157:H7.

Strains of enterohemorrhagic Escherichia coli (EHEC) serotype O157:H7 produce under stress copious amounts of exopolysaccharide (EPS) composed of colanic acid (CA). Studies were performed to evaluate the association of production of CA with survival of EHEC under adverse environmental conditions. A CA-deficient mutant, M4020, was obtained from a CA-proficient parental strain, E. coli O157:H7 W6-13, by inserting a kanamycin resistance gene cassette (kan) into wcaD and wcaE, 2 of the 21 genes required for CA biosynthesis. M4020 was defective in CA production as determined from the ratio of uronic acid to protein (UA/P) of cells grown from 1 to 4 days at 25 degrees C on minimal glucose agar (MGA), MacConkey agar, and sorbitol-MacConkey agar, and by colony morphology on MGA. The results of stress treatment revealed that M4020 was substantially less tolerant to acid (pH 4.5 and 5.5) and heat (55 and 60 degrees C) in comparison to W6-13, indicating that CA confers on E. coli O157:H7 a protective effect from the environmental stresses of acid and heat.

Escherichia coli O157↗
PubMed2001

Chlorine inactivation of Escherichia coli O157:H7 in water.

Six human isolates of Escherichia coli O157:H7 and E. coli (ATCC 11229) were used to determine the concentrations of free chlorine and exposure times required for inactivation. Free chlorine concentrations of 0.25, 0.5, 1.0, and 2.0 ppm at 23 degrees C were evaluated, with sampling times at 0, 0.5, 1.0, and 2.0 min. Results revealed that five of six E. coli O157:H7 isolates and the E. coli control strain were highly susceptible to chlorine, with >7 log10 CFU/ml reduction of each of these strains by 0.25 ppm free chlorine within 1 min. However, comparatively, one of the seven strains was unusually tolerant to chlorine at 23 degrees C for 1 min, with a 4-, 5.5-, 5.8-, and >5.8-log CFU/ml reduction at free chlorine concentrations (ppm) of 0.25, 0.5, 1.0, and 2.0. respectively. Based on these studies most isolates of E. coli O157:H7 have no unusual tolerance to chlorine; however, one strain was exceptional in being recovered after 1-min of exposure of 10(7) CFU/ml to 2.0 ppm of free chlorine. This isolate may be a useful reference strain for future studies on chlorine tolerance of E. coli O157:H7.

Chlorine↗
PubMed2001

Evaluation of universal preenrichment broth for growth of heat-injured pathogens.

Universal preenrichment broth (UPB) was developed to enable enrichment of injured foodborne pathogens of different genera simultaneously in lieu of having to undergo separate simultaneous enrichment cultures for subsequent detection or isolation of each pathogen. Enrichment conditions in UPB for growth of injured pathogens to populations that will enable pathogen detection by rapid immuno-based or polymerase chain reaction (PCR)-based assays have not been defined. Hence, studies were done to determine recovery and growth rates of heat-injured Escherichia coli O157:H7, Salmonella enterica ser. Typhimurium, Salmonella enterica ser. Enteritidis. and Listeria monocytogenes in UPB. Bacterial cells were heat injured in tryptic phosphate broth at 57.2 degrees C and inoculated at populations of ca. 0.17 to 63 injured cells per ml with raw ground beef, fresh chicken, lettuce, and environmental sponge samples. Enrichment cultures were sampled at 1, 2, 3, 4, 5, 6, and 24 h at 37 degrees C postinoculation, and pathogens were enumerated on appropriate selective media. Results revealed that recovery and growth of pathogens during the first 6 h of enrichment were not sufficient to ensure adequate numbers of bacteria (> 10(3) CFU/ ml) for detection by most immunoassays or PCR assays. Cells often required 3 to 4 h for recovery before growth was initiated. Salmonella Typhimurium, Salmonella Enteritidis, E. coli O157:H7, or L. monocytogenes cell populations in enrichment cultures with ground beef or lettuce at 6 h were 0.5 to 2.9 log10 CFU/ml. At 24 h of incubation, cell counts of enrichment samples for the three pathogens from all food and environmental sponge samples ranged from 4.0 to 8.3 log10 CFU/ml. Enrichment in UPB at 37 degrees C of foods or environmental sponge samples containing heat-injured cells of Salmonella Typhimurium, Salmonella Enteritidis, E. coli O157:H7, or L. monocytogenes reliably provides at 24 h of incubation-but not at 6 h-sufficient cell populations for detection by rapid immunoassay or PCR assay procedures that can detect at least 4 log10 CFU/ml. These results raise questions regarding the sensitivity of rapid detection methods that employ an abbreviated enrichment protocol of 6 h or less.

Animals↗
PubMed2001

Fate of salmonellae in calcium-supplemented orange juice at refrigeration temperature.

Recent outbreaks of salmonellosis associated with orange juice have raised interest concerning the survival and growth of Salmonella in juice supplemented with calcium. A study was done to determine the influence of various calcium supplements on the survival of salmonellae in orange juice held at 4 degrees C for up to 32 days. Isolates of Salmonellal Muenchen (inoculum 1), Salmonella isolates from humans and animals (inoculum 2), and Salmonella isolates from produce outbreaks (inoculum 3) were inoculated into pasteurized orange juices with pH values ranging from 3.96 to 4.19 and containing 350 mg of calcium per 240-ml serving (1.46 mg of calcium/ml). Populations of Salmonella declined rapidly in juice containing calcium lactate (CaL), with counts decreasing from 4.86 log10 CFU/ml to < 1 log10 CFU/ml within 16 days, regardless of the Salmonella serotypes present in inoculum. Counts decreased from 4.89 log10 CFU/ml to < 1 log10 CFU/ml in orange juice supplemented with CaL and tricalcium phosphate (TCP) within 30 days. These reductions were significantly (P < or = 0.05) higher than those of the control (no calcium added), in which Salmonella populations decreased 3.19 +/- 0.20 log10 CFU/ml over 32 days. Populations in orange juice containing TCP or calcium citrate (CC) declined 1.34 +/- 0.20 log10 CFU/ml and 1.96 +/- 0.20 log10 CFU/ml, respectively, over 32 days. These counts were significantly higher than respective control counts in juice stored for 32 days. Populations of Salmonella of inoculum 3 inoculated into juice containing calcium citrate malate (CCM) were significantly higher than in the control. Higher numbers of cells in inoculum 3 also survived compared to numbers of cells of inocula 1 or 2 in juice supplemented with CCM. This study reveals that the form of calcium used to supplement orange juice influences the ability of salmonellae to survive.

Beverages↗