Browsing by Author "Sumner, Susan S."

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    Acceptability and Shelf-Life of Fresh and Pasteurized Crab Meat Stored Under Different Environmental Conditions
    Tyler, Carla Gutierrez (Virginia Tech, 2009-02-02)
    Crab meat is important to the economy of coastal Virginia. The objectives of this study were to complete a shelf-life study on two different packaging styles of fresh crab meat and to test the inhibition capabilities of Carnobacterium piscicola against the pathogen, Listeria monocytogenes. In a shelf-life study, a 12 ounce food grade polyethylene traditional snap-lid container of fresh crab meat was compared to an 8 ounce SimpleStep® trays with Cryovac™ film of equally fresh crab meat sealed with 10,000 cc/m2/24hr oxygen transmission rate (OTR) film. Eleven g samples were used for the microbial shelf-life study conducted at 4°C for 12 days. Aerobic plate counts of crab meat indicated microbial growth from the SimpleStep® trays with Cryovac™ film in 10,000 cc/m2/24hr OTR versus the polyethylene snap-lid was not significant (P>0.05). In objective two, 25 g samples of fresh and pasteurized blue crab (Callinectes sapidus) meat were inoculated with 0.1ml of each, C. piscicola and L. monocytogenes. Three different concentrations of the inoculation levels were studied on select days at both 4°C and 10°C. Microbial spoilage was defined as 107 CFU/g. In fresh crab meat, at both 4°C and 10°C, crab meat spoilage occurred at 7 days or less. In the pasteurized crab meat, at 4°C and 10°C, spoilage did not occur prior to 26 days, and studies were terminated at 28 days of storage. The growth of the two organisms in fresh crab meat was found to be significant for the differing concentration levels and sampling days (P<0.05). The growth of the two organisms in pasteurized crab meat was significant for different concentration levels, sampling days and temperature (P<0.05). In both fresh and pasteurized crab meat, regardless of the inoculation ratios, the L. monocytogenes and C.piscicola followed similar growth trends, but L. monocytogenes was higher in the 2:2 CFU/g concentration and lower at the 6:2 CFU/g concentration level. Although C. piscicola did not completely inhibit L. monocytogenes growth at any concentration ratio, some inhibition was observed.
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    The Affects of Explosively and Electrically Generated Hydrodynamic Shock Waves on the Bacterial Flora of Beef and Poultry
    Lorca, Tatiana Andrea (Virginia Tech, 2002-07-19)
    The affects of hydrodynamic shock wave treatment on the bacterial flora of raw beef and poultry were evaluated. Hydrodynamic shock waves were generated in an aqueous treatment medium by either the detonation of two types of explosive charges (explosively-generated hydrodynamic shock waves [EHSW]) (a binary or a molecular explosive) or by electrical discharge (high voltage arc discharge Hydrodyne (TM [HVADH; Hydrodyne, Inc.]). A variety of sample types (whole steaks, ground beef, a water and ground beef slurry) were used to determine the lethality affects of EHSW on cells of the marker microorganism Listeria innocua suspended in a simple broth medium. These sample types were used in order to evaluate the affects of the process not only on the surface, but throughout the bulk of the samples in order to determine whether EHSW could also be used as a non-thermal alternative to reduce the bacterial flora of non-intact or ground meats. The levels of psychrotrophic, lactic, and coliform populations on the surface of whole eye of round steaks submitted to EHSW processing did not differ (P> 0.05) from those of untreated whole eye of round steaks. Parameters expected to influence the nature, magnitude, and propagation of the hydrodynamic shock wave were also varied and evaluated in order to determine which individual parameter or combination of parameters affected the bactericidal potential of EHSW or HVADH processing. Treatment with EHSW failed (P > 0.05) to produce lethality effects on the psychrotrophic, lactic, and coliform populations of ground beef, regardless of the composition and mass of explosive used, the number of successive EHSW treatments used, the relative distance between the explosive charge and the top surface of the sample, or the temperature of the water used in the treatment chamber. EHSW processing did not change (P >0.05) the bacterial population of treated ground beef samples when compared to untreated controls during a five day refrigerated storage study. No lethality effects were observed (P >0.05) in ground beef samples treated by HVADH when samples were subjected to one, two, or three successive HVADH treatments. Minimal penetration of surface inoculated bacteria was observed for both beef steaks and boneless skinless chicken breasts subjected to EHSW and HVADH, respectively. In EHSW-treated beef eye of round steaks, marker bacteria were detected within the first 300 um of tissue below the inoculated surface, 50-100 um beyond the depth of untreated surface inoculated steaks. In HVADH-treated boneless skinless chicken breasts, marker bacteria were detected within the first 200 um below the inoculated surface, 50-100 um beyond the depth of untreated surface inoculated boneless skinless chicken breasts. This suggests that although no difference in the bacteriological populations was observed between EHSW treated, HVADH treated, and untreated control samples of whole steaks (and ground beef treated with both HVADH and EHSW), HVADH and EHSW treatments affect the movement of surface bacteria. United States Department of Agriculture (USDA) guidelines suggest intact beef steaks be cooked to achieve a cooked color appearance on the surface and raw poultry be cooked to an internal temperature of 77° C to inactivate the pathogens Escherichia coli O157:H7 and salmonellae which are of concern in beef and poultry, respectively. By following these guidelines during proper cooking, consumers achieve thermal inactivation of these pathogens. Since the movement of the marker bacterium observed in treated steaks and boneless skinless chicken breasts was minimal, proper cooking of the products would be expected to inactivate vegetative bacterial cells at this depth. Therefore, EHSW and HVADH treated whole beef steaks and boneless skinless chicken breasts would not be expected to pose a bacterial hazard if the products were properly cooked.
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    Antibacterial Activity of Hydrogen Peroxide Against Escherichia Coli O157:H7 and Salmonella Spp. in Fruit Juices, Both Alone and in Combination With Organic Acids
    Schurman, John Jackson (Virginia Tech, 2001-07-18)
    The antibacterial efficacy of hydrogen peroxide treatments in four fruit juices was determined. Preservative free apple cider, white grape, and purple grape juice were inoculated with ~ 6.4 log CFU/ml of a five strain, acid adapted, nalidixic acid resistant E. coli O157:H7 cocktail. Orange juice was inoculated with a comparable Salmonella spp. cocktail. In the first study, 0.017% and 0.012% H₂O₂ was added in combination with 0.1% and 0.3% of the dominant organic acid (OA) to 4°C and 25°C juices, with samples taken each day for 21 days. H₂O₂ was a significant factor in all juices (p < 0.05) except white grape (lack of data), and both 0.017% H₂O₂ treatments reduced counts in apple cider, orange juice, and white grape to undetectable numbers within 48 hrs as cultured on tryptone soy agar + 0.05% nalidixic acid (TSAN). Treatments in purple grape juice were less effective overall, and more dependent on OA concentration (p < 0.001) than H₂O₂. There were instances where bacterial survival in apple cider, purple grape, and orange juice continued for 21 days after treatment, and sometimes outlasted the control. These occurrences were dependent on temperature (25°C) and H₂O₂ (0.012%), but not on OA. However, OA concentration was a significant factor (p < 0.05) overall in apple cider and purple grape juice, but not in orange juice. In the second study, 0.015% and 0.03% H₂O₂ was added to 10, 25, and 40°C apple cider and orange juice inoculated with 6.4 log CFU/ml E. coli O157:H7 and Salmonella spp. respectively. Only 0.03% H₂O₂ was effective in reducing counts to undetectable numbers in both juices. However, both temperature and H₂O₂ were significant factors (p < 0.0001) in bacterial destruction, with 0.03% H₂O₂ at 40°C giving undetectable numbers at ≤ 3 and ≤ 6 hours in orange juice and apple cider respectively. It has been demonstrated that at ~ ≥ 0.017%, H₂O₂ can provide a 5 log reduction of these pathogens in fruit juice. Increasing temperature and organic acid concentration can improve its rate of effectiveness in certain juices. However, sensory concerns may negate its use in some products.
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    Clostridium botulinum toxin development in refrigerated reduced oxygen packaged Atlantic croaker (Micropogonias undulatus)
    Rheinhart, Courtney Elizabeth (Virginia Tech, 2007-04-30)
    The purpose of this study was to determine the effects of storage temperature and film oxygen transmission rate (OTR) on toxin development by Clostridium botulinum in refrigerated raw vacuum packaged croaker fillets, and to determine if toxin development precedes microbiological and/or organoleptic spoilage. Raw croaker fillets were vacuum packaged in oxygen-permeable films (OTR of 10,000 cc/m2/24hr or 3,000 cc/m2/24hr) and stored at either 4ºC or 10ºC. Type 83F, 17 Type B, Beluga, Minnesota, and Alaska nonproteolytic strains of C. botulinum were used to inoculate fish prior to vacuum packaging. At both temperatures, microbial spoilage preceded toxin production in fillets vacuum packaged in both film types. At 4ºC microbial spoilage occurred after approximately 7 days for fillets vacuum packaged in the 10,000 cc/m2/24hr OTR film and after 8 days for fillets vacuum packaged in the 3,000 cc/m2/24hr OTR film. However, toxin was not detected until day 8. At 10ºC microbial spoilage occurred after approximately 3 days for fillets vacuum packaged in the 10,000 cc/m2/24hr OTR film, while toxin production occurred on day 5. For fillets vacuum packaged in the 3,000 cc/m2/24hr OTR film microbial spoilage occurred after 4 days. However toxin production did not occur until day 6. In contrast, at both temperatures toxin production preceded or coincided with organoleptic spoilage in fillets vacuum packaged in both film types. At 4ºC organoleptic spoilage occurred after 10 days for fillets packaged in the 10,000 cc/m2/24hr OTR film and after 9 days in the 3,000 cc/m2/24hr OTR film, while toxin production occurred on day 8. At 10ºC organoleptic spoilage occurred after 6 days for fillets packaged in the 10,000 cc/m2/24hr OTR film, and toxin was detected on day 5. For fillets packaged in the 3,000 cc/m2/24hr OTR film and stored at 10ºC, organoleptic spoilage occurred after 6 days, while toxin production occurred on day 6. Although toxin production preceded or coincided with organoleptic spoilage in both film types, this may have been because samples were presented on ice, which could have masked potential odors. This study shows that there are not significant differences between these film types when it comes to microbial and organoleptic spoilage. Therefore lower OTR films, such as 3,000 cc/m2/24hr film, may be used to vacuum package Atlantic croaker.
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    Comparative Study of Semisynthetic Derivative of Natamycin and the Parent Antibiotic on the Spoilage of Shredded Cheddar Cheese
    Suloff, Eric Charles (Virginia Tech, 1999-11-12)
    The polyene macrolide antibiotic natamycin (Antibiotic A-5283) is commonly used to retard the growth of surface molds on various cheese varieties. Natamycin is commonly applied to the surface of cheese by dipping or spraying, using an aqueous dispersion containing 200 to 300 ppm of the additive. The large molecular weight of natamycin, 666 g/mol, and conjugated double bond structure causes it to be extremely insoluble in water and most food grade solvents. The inability to apply natamycin in true solution creates void non-treated areas on the food surface. These non-treated areas promote the growth of fungal organisms. A water soluble N-alkyl semisynthetic derivative of natamycin was synthesized by the Michael addition reaction of the parent with a N-substituted malemide. A comparative study investigating the effectiveness of the semisynthetic derivative of natamycin and the parent antibiotic in suppressing mold growth on one month aged shredded Cheddar cheese modified atmosphere packaged (MAP) was performed. A 20 ppm natamycin treatment effectively suppressed visible mold growth (<104 CFU/g) in MAP samples for up to 30 days after opening. The 20 ppm semisynthetic derivative performed similarly to the 10 ppm natamycin treatment in retarding mold growth. Visible mold growth did not occur for these treatments in MAP samples until 20 days after opening. Analysis of storage conditions revealed that an outgrowth of mold in shredded cheese occurred in MAP packages stored longer than 15 days. This bloom in mold growth was attributed to the degradation of natamycin and the semisynthetic derivative throughout storage. The stability and degradation of natamycin and the derivative were monitored throughout the study. Antibiotic concentration on the cheese surface was quantified by molecular absorption spectrometry. Results from this study showed, heavily contaminated samples caused the rate and loss of natamycin and the derivative to increase. Antibiotic concentration decreased at a similar rate in MAP and open package conditions. Natamycin and derivative were found to have similar degradation properties.
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    Development of Immunomagnetic Capture (IMC) Based Techniques For the Detection of Salmonella in Poultry Carcass Rinse Fluid
    Sharp, Jennifer M. (Virginia Tech, 2002-07-12)
    Current detection methods require at least one 24-48 hour enrichment step for the detection of Salmonella. This poses a problem because product often needs to be shipped before microbial contamination levels can be adequately ascertained. Therefore, the need for more rapid methods of Salmonella detection becomes apparent. The purpose of this thesis was to determine if an immunologically-based method, Immunomagnetic Capture(IMC) -ELISA and molecular-based detection methods, PCR and Taqman® PCR employing IMC without enrichment, could detect at least 102 cfu/ml of S. Typhimurium in broiler carcass rinse fluid (CRF) samples. IMC-ELISA, IMC-PCR, and IMC-Taqman® PCR were initially tested using 0 to 106 cfu/ml of pure culture S. Typhimurium. Each detection method was tested using artificially contaminated CRF samples. Finally, standardized IMC-ELISA, IMC-PCR, and IMC-Taqman® PCR methods were tested using commercial CRF samples. Salmonella concentrations were verified using a traditional plate method. IMC-ELISA produced consistent results when detecting at least 104 to 106 cfu/mL of pure culture S. Typhimurium. IMC-ELISA was not able to produce repeatable results when testing artificially contaminated CRF samples. S. Typhimurium was not detected in commercial CRF samples which by virtue of direct plating on XLT-4 were found to contain essentially no Salmonella (<1 cfu/ml). IMC-PCR was able to consistently detect 102 cfu/ml, whereas IMC-Taqman® PCR was able to detect 101 cfu/ml of pure culture S. Typhimurium. IMC-PCR, required four hours to complete, and it consistently detected 104 cfu/ml of S. Typhimurium in artificially contaminated CRF samples. IMC-Taqman® PCR took 3 hours to perform and was able to detect 103 cfu/ml S. Typhimurium in artificially contaminated CRF samples. The sensitivity, as well as the decreased time requirements of these detection methods, would suggest their usefulness in a commercial processing setting.
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    Effect of Alternative Household Sanitizing Formulations Including: Tea Tree Oil, Borax, and Vinegar, to Inactivate Foodborne Pathogens on Food Contact Surfaces
    Zekert, Ashley Elizabeth (Virginia Tech, 2009-11-09)
    Current trends indicate that American consumers are increasingly selecting products that they believe to be environmentally friendly or "natural." In the kitchen, this trend has been expressed through greater desire for using alternative or "green" sanitizers instead of bleach or other common chemical sanitizers. The purpose of this work was to evaluate the effectiveness of one suggested alternative, tea tree oil, as a food contact surface sanitizer. Three foodborne bacterial pathogens (Listeria monocytogenes N3-031 serotype 1/2a, Escherichia coli O157:H7 strain E009, and Salmonella Typhimurium ATCC 14028) were applied separately onto three different food contact surfaces (high density polyethylene, glass, and Formica® laminate). Tea tree oil (TTO), borax, and vinegar (5% acetic acid) were applied individually as well as in combination for a total of seven treatment solutions. In addition, household bleach (6.15% sodium hypochlorite), sterile reverse osmosis (RO) water, and no applied treatment were used as controls. Treatments were tested using an adaptation of the Environmental Protection Agency DIS/TSS-10 test method, whereby each contaminated surface was treated with 100 µl of test solution and held for 1 min followed by submersion in neutralizing buffer and microbiological plating. Samples (0.1 ml) were plated onto TSA and incubated at 35°C for 48 h prior to colony counting. Bleach reduced microbial populations significantly with greater than 5-log reduction reported for all surfaces (Formica® laminate, glass, and HDPE), against E. coli O157:H7, L. monocytogenes, and S. Typhimurium. TTO produced reductions between four and five logs for E. coli O157:H7, L. monocytogenes, and S. Typhimurium and was not statistically different from the vinegar treatment (P>0.05). All combination recipes, including the borax treatment, failed to produce reductions in microbial populations at levels considered to be appropriate for food contact surface sanitizers. Surface type did not play a significant role in the effectiveness of the treatment (P>0.05). Although TTO and vinegar did reduce pathogen populations on surfaces, reductions were not sufficient enough to be considered an equally effective alternative to household bleach.
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    Effect of Evaporative Cooling, Fat Content and Food Type on Pathogen Survival during Microwave Cooking
    Hix, April (Virginia Tech, 2000-07-28)
    Due to the rapid nature of microwave heating, the microbiological safety of foods prepared in the microwave has been in question for several years. Because foods are heated from the inside out and are strictly governed by their own internal properties such as ionic content, moisture level and specific heat, work must be done to further master control of such properties so microwave cooking can be more predictable, controlled and ensure control pathogens. This study concentrated on the effect of fat content, evaporative cooling and food type on the rate of food borne pathogen survival rates in microwave heated foods. Foods investigated in this study included fresh, raw broccoli spears; a regular, whole muscle breaded chicken patty and a fat free, breaded, formed chicken patty; and raw ground beef patties at three differing fat percentages. All foods were tested in triplicate. A Sharp® 1000W Light-Duty Commercial Microwave Oven was used to treat inoculated samples according to their recommended cooking times. Two sets of samples were treated, one wrapped with Saran™ Wrap and the other without wrap. F-values were determined for each product. Raw ground beef patties at fat contents of 30%, 15% and 7%, heated for the same time had F-values ranging from 0.03 to 126.20. The lower the fat content, the lower the lethality. Regular and fat free chicken tenders had similar patterns. F-values for fresh broccoli indicted that vegetative pathogens survived the recommended microwave process. Covering in Saran™ Wrap had some preventive effect on evaporative cooling depending on the food tested and significantly (p < 0.05) increased most F-values. Inoculated pack studies were performed in triplicate on each food with Listeria monocytogenes, Salmonella and Escherichia coli O157:H7. Survival was determined by presence or absence of growth of each pathogen after enrichment. Listeria monocytogenes survived in all samples except for the 30% fat ground beef patties. The Salmonella species had a lower survival rate; however, it was still present in uncovered 15% fat ground beef, covered 7% fat ground beef, uncovered chicken patties (both types) and in all broccoli samples tested. E. coli O157:H7 survived in all samples except the 30% fat ground beef samples. Results indicate that higher fat contents seem to ensure lower rates of pathogen survival. This was especially true for the raw ground beef, which had received no prior processing other than the grinding of the whole muscle. There were fewer survival differences in the preprocessed, frozen chicken patties. Both were shown to support no pathogen survival in covered samples, except the fat free chicken patties. Listeria monocytogenes was shown to consistently survive the suggested cooking time in these samples. This is consistent with expectations that fat free food samples would display more survival than regular fat samples. Overall, covering samples with Saran™ had little effect on pathogen survival rates. There were survival differences in some covered and uncovered samples consistent with expectations that covered samples would show less survival than uncovered, but further work including more samples would be necessary to ensure that the covered or uncovered variable made the true difference in pathogen survival. Finally, broccoli demonstrated consistent pathogen survival in all categories of testing. This indicates microwave oven prepared vegetables could be a prime source of pathogen transmission to consumers. Further work needs to concentrate on determining the correct processing times and parameters that need to be met to ensure safe food.
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    Effect of Fat Content and Food Type on Heat Transfer during Microwave Heating
    Gunasekaran, Nishkaran (Virginia Tech, 2002-08-08)
    Microwaves heat food rapidly and foods are prepared in less time. However, due to non-uniform heating nature of microwave cooking, there exists a serious concern over complete elimination of pathogens in the food. There has been an increase in interest to accurately understand the behavior of different food materials in a microwave field and microbial inactivation during microwave cooking. Recent research showed that fat content in muscle food plays an important role in microbial inactivation by increasing the inactivation level with an increase in the fat level. It was also demonstrated that muscle food heats up differently than a vegetable food product. Cooking food in a microwave oven either by covering the food container or not results in significantly different temperature profiles. The current research attempts to use modeling techniques to analyze impact of these factors on microwave heating. Mathematical modeling is faster, easier and economically better than actual experiments in determining heating behavior of a microwave-cooked food. Though modeling cannot completely replace actual experiments, it can be used as a tool to understand the effects of various factors influencing the microwave cooking. A factor that is highly important during microwave processing is dielectric properties of the material. The interaction of microwave with the food is mainly based on its dielectric properties, which can change with temperature. Therefore, determination of dielectric properties of food with respect to temperature becomes critical. The current research project has two parts. One to determine the dielectric properties of food being tested and another is to employ mathematical modeling techniques to analyze the effect of fat content, food type and the effect of cooking food by covering the bowl using the lid and not covering bowl. Dielectric properties of ground beef patties at 4%, 9%, 20% fat levels and frozen broccoli were determined using an open-ended, 3.6 mm diameter, semi-rigid coaxial line with copper conductors, connected to a network analyzer. The properties were determined at various temperatures. Foods were measured in triplicate. Results showed that dielectric constant and dielectric loss factor of low fat ground beef were higher than that of high fat level ground beef. In addition, the dielectric properties of florets were lower than that of stem parts for frozen broccoli. A 1,200W, household type microwave oven was used in this study to heat the food. Food was placed in a microwave-safe glass bowl and cooked for 120 seconds. One headspace and three internal temperature measurements were recorded for every 0.6 seconds. Five replications were performed. Finite element method was used as modeling technique and temperatures were predicted. Experimental and predicted temperature values were compared. Results showed that the model used in the study was more suitable for modeling the uncovered cooking than covered cooking process. Modeling results also revealed that high fat ground beef patties reached higher temperature than low fat patties. In high fat meat products, fat content also contributed to increase in temperature during microwave heating. In vegetable products and low fat meat food, moisture content is mainly responsible for microwave heating. A more extensive study on critical fat level above which fat content helps in increasing temperature is needed. In addition, inclusion of steam properties in the headspace for modeling the covered cooking is recommended.
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    Effect of hydrogen peroxide and other protease inhibitors on Cryptosporidium parvum excystation and in vitro development
    Kniel, Kalmia E.; Sumner, Susan S.; Pierson, Merle D.; Zajac, Anne M.; Hackney, Cameron Raj; Fayer, Ronald; Lindsay, David S. (American Society of Parasitology, 2004-08)
    This study was undertaken to observe the effects of hydrogen peroxide on Cryptosporidium parvum oocysts with respect to protease activity in comparison to known protease inhibitors. In assessing the possible mechanisms of action of hydrogen peroxide, treatment effectiveness was analyzed using 3 assays and the potential roles of proteases and cations were considered. Treatment of C. parvum oocysts with hydrogen peroxide inhibited protease activity up to 50% compared with untreated controls. Treatment of oocysts with chemicals that affect sulfhydryls, including N-ethylmaleimide and dithiolthreitol, inhibited protease activity by > 90%. Treatment of oocysts with these chemicals, along with the protease inhibitors, phenylmethylsulfonyl fluoride (PMSF), ethylenediamine-tetraacetic acid, and cystatin, inhibited protease activity as well as in vitro excystation and infection in a cell culture assay. Several mechanisms may result in the successful inhibition of infection and excystation by hydrogen peroxide treatment, including: oxidation of oocyst wall proteins or lipids, chelating of cations necessary for infection, or hydroxyl radical-induced DNA damage to sporozoites, or both.
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    Effect of Metabolic Enzymes on Amylopectin Content and Infectivity of Cryptosporidium parvum
    Hartman, Angela Danielle (Virginia Tech, 2006-11-20)
    Amylopectin granules in Apicomplexan protozoa are hypothesized to be used as an energy source to aid the parasites in surviving in the environment allow latent stages to excyst and release infective stages, allow them to be mobile, invade host cells, and to continue their life cycle. The objective of this project was to determine if parasite glycolytic enzymes: alpha-amylase, amyloglucosidase, enolase, lactate dehydrogenase, and phosphorylase could be used to decrease amylopectin stores and subsequently infectivity of Cryptosporidium parvum oocysts/sporozoites in both fresh oocysts and stored oocysts. In addition, glycolytic enzymes and substrates: glucose, glucose-1-phosphate, and glycogen synthase were investigated to determine if they can be used to increase amylopectin stores and thus increase infectivity to aid in detection/storage of oocysts. Oocysts of Cryptosporidium parvum were suspended in 1mg/ml glycolytic enzymes or substrates (except glucose - 0.05M and glycogen synthase - 1U/ml) and electroporated. Oocysts were incubated at 37°C for one hour to allow treatments to react with amylopectin followed by incubation on HCT-8 cells for 24 hours for infection. Real-time PCR and immunohistochemistry were performed to determine the effect of the enzymes on infectivity. An amylopectin assay and excystation assay was performed to determine if the enzymes degraded amylopectin and if decreased amylopectin reduced excystation. Alpha amylase and amyloglucosidase had the greatest impact on reducing both amylopectin and infectivity of fresh oocysts with reductions of 99.5% and 99.1% in infective oocysts, respectively (P<0.05). These results suggest that amylopectin may be an important factor in infection, although further research is needed. In stored oocysts, enzymes significantly reduced amylopectin content but not infectivity. In fresh oocysts, amylopectin content was correlated to excystation and infectivity with a decrease in amylopectin correlating to decreased excystation and infectivity. In contrast, there was no direct correlation for stored oocysts. When glucose, glucose-1-phosphate, or glycogen synthase was used to increase infectivity, results show that glycogen synthase had little effect, but glucose and glucose-1-phosphate significantly increased amylopectin content, excystation, and infectivity. In conclusion, amylopectin may be an important polysaccharide store of Cryptosporidium parasites to cause infection by allowing excystation of the oocysts to release infective sporozoites.
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    The effect of milkfat melting properties on chemical and physical properties of 20% reformulated cream
    Scott, Lisa Lenore (Virginia Tech, 1999-09-17)
    Skim, sweet buttermilk, and butter derived aqueous phase components were used to re-emulsify low-melt and medium-melt fraction butteroils to yield 20% milkfat creams. The implications of separation temperature in obtaining components, melting point characteristics, and formulation on the chemical and physical properties of reformulated and natural creams were analyzed. Transmission electron microscopy indicated that both reformulated and natural creams were oil-in-water emulsions, demonstrating lipid globules surrounded by surface material. Chemical analysis of components proved that sweet buttermilk and butter-derived aqueous phase components had significantly higher (p less than or equal to 0.01) amounts of cholesterol and phospholipid than skim milk, resulting in creams formulated with sweet buttermilk and butter-derived aqueous phase creams having significantly higher (p less than or equal to 0.01) amounts of cholesterol and phospholipid than creams formulated with skim milk. Butter-derived aqueous phase had higher (p less than or equal to 0.01) amounts of lipid, cholesterol, and phospholipid than sweet buttermilk. However, skim component had higher (p less than or equal to 0.01) amounts of protein than butter-derived aqueous phase. When compared to natural creams, creams consisting of sweet buttermilk and butter-derived aqueous phase components had similar amounts of total phospholipid and amount of phospholipid adsorbed to lipid globules than creams consisting of skim component. Creams consisting of skim component had higher (p less than or equal to 0.01) amounts of protein than natural cream. Reformulated creams having low-melt fraction butteroil had higher (p less than or equal to 0.01) amounts of cholesterol. For reformulated creams, creams processed from components obtained by 49oC separation had significantly higher (p less than or equal to 0.01) amounts of cholesterol than like creams manufactured from 55oC separation components. Creaming stability, viscosity, feathering, and sensory quality of reformulated and natural creams were analyzed over a 13 day storage period at 3.3oC. Formulation, separation temperature, or melting point characteristics did not significantly (p greater than 0.01) affect creaming stability of reformulated and control creams homogenized at 13.6/3.4 MPa. The day within storage period, however, was a significant factor (p less than or equal to 0.01) in determining creaming stability of reformulated and natural creams. All creams displayed typical non-Newtonian behavior at 7oC, displayed by hysteresis curves in which viscosity decreased as shear rate increased. Formulation and separation temperature used to obtain components did not have a significant (p greater than 0.01) effect on viscosity; however, all creams formulated with medium-melt fraction butteroil had significantly (p less than or equal to 0.01) higher apparent viscosity values than creams with low-melt fraction butteroil at shear rate 692.48 s-1 and at 1384.96 s-1 and 2769.92 s-1 for creams formulated with skim component. Regardless of formulation, separation temperature, and melting point characteristics, all creams feathered in a pH range of 4.70-5.09. Reformulated and natural creams met sensory quality specifications as determined by the In/Out Method of Specification, except for creams formulated with skim milk and low-melt fraction butteroil which were characterized as having oxidized flavors. Creams formulated with buttermilk and butter derived aqueous phase had more comparable physical properties to natural creams than skim milk creams.
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    Effect of Ozone and Ultraviolet Irradiation Treatments on Listeria monocytogenes Populations in Chill Brines
    Dev Kumar, Govindaraj (Virginia Tech, 2008-11-19)
    The efficacy of ozone and ultraviolet light, used in combination, to inactivate Listeria monocytogenes in fresh (9% NaCl, 91.86% transmittance at 254 nm) and spent chill brines (20.5% NaCl, 0.01% transmittance at 254 nm) was determined. Preliminary studies were conducted to optimize parameters for the ozonation of "fresh" and "spent" brines. These include diffuser design, comparison of kit to standard methods to measure residual ozone, studying the effect of ozone on uridine absorbance and determining presence of residual listericidal activity post ozonation. An ozone diffuser was designed using 3/16 inch PVC tubing for the ozonation of brines. The sparger was designed to facilitate better diffusion and its efficiency was tested. The modified sparger diffused 1.44 ppm of ozone after 30 minutes of ozonation and the solution had an excess of 1 ppm in 10 minutes of ozonating fresh brine solution (200ml). Population levels of L. monocytogenes were determined at various time intervals post-ozonation (0, 10, 20, 60 min) to determine the presence of residual listericidal activity. The population post ozonation (0 minutes) was 5.31 Log CFU/ml and was 5.08 Log CFU/ml after a 60 minute interval. Therefore, residual antimicrobial effect was weak. Accuracy of the Vacu-vial Ozone analysis kit was evaluated by comparing the performance of the kit to the standard indigo colorimetric method for measuring residual ozone. The kit was inaccurate in determining residual ozone levels of spent brines and 1% peptone water. Uridine was evaluated as a UV actinometric tool for brine solutions that were ozonated before UV treatment. The absorbance of uridine (A262) decreased after ozonation from 0.1329 to 0.0512 for standard 10 minutes UV exposure duration. Absorbance of uridine was influenced by ozone indicating that the presence of ozone may hamper UV fluence determination accuracy in ozone-treated solutions. Upon completion of diffuser design and ozone/UV analysis studies, the effect of ozone-UV combination on L. monocytogenes in fresh and spent brines was evaluated. Ozonation, when applied for 5 minutes, caused a 5.29 mean Log reduction while 5 minutes of UV exposure resulted in a 1.09 mean Log reduction of L. monocytogenes cells in fresh brines. Ten minutes of ozonation led to a 7.44 mean Log reduction and 10 minutes of UV radiation caused a 1.95 mean Log reduction of Listeria in fresh brine. Spent brines required 60 minutes of ozonation for a 4.97 mean Log reduction in L. monocytogenes counts, while 45 minutes resulted in a 4.04 mean Log reduction. Ten minutes of UV exposure of the spent brines resulted in 0.30 mean Log reduction in Listeria cells. A combination of 60 minutes ozonation and 10 minute UV exposure resulted in an excess of 5 log reduction in cell counts. Ozonation did not cause a sufficient increase in the transmittance of the spent brine to aid UV penetration but resulted in apparent color change as indicated by change in L*a*b* values. Ozonation for sufficient time had considerable listericidal activity in fresh brines and spent brines and when combined with UV treatment, is effective reducing L. monocytogenes to undetectable levels in fresh brines.
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    The Effect of Sorbic Acid on the Survival oOf Escherichia coli 0157:H7, Salmonella, Listeria monocytogenes, and Staphylococcus aureus on Shredded Cheddar and Mozzarella Cheese
    Roberts, Alison K'Ann (Virginia Tech, 2002-12-15)
    The objective of this study was to determine the effectiveness of sorbic acid in inhibiting Escherichia coli 0157:H7, Salmonella spp., Listeria monocytogenes, and Staphylococcus aureus on shredded cheddar and mozzarella cheese over 70 days storage. Samples of cheese were inoculated and placed into bags with a sorbic acid (0, 0.1, 0.15, 0.2 and 0.3 %) and anti caking agent mixture and stored at 10 °C. Each variable was enumerated after 0,14,28,42,56, and 70 days of storage. Survival of E. coli 0157:H7 showed no significant difference from control in either cheese. There were significantly lower Salmonella counts for days 14 to 42 on mozzarella cheese. No significant differences in survival were found for cheddar cheese. There were significantly lower counts noted in L. monocytogenes, and S. aureus in mozzarella. Though no significant differences were found over time in the cheddar, most of the sorbate concentrations exhibited lower counts than control on days 14 and 28. Overall, in the presence of sorbic acid there was a more rapid decline in numbers of each test organism, especially against L. monocytogenes, and S. aureus for both high and low moisture cheeses.
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    The Effectiveness of Potassium Lactate and Lactic Acid Against Campylobacter Species and Psychrotrophic Bacteria
    Rasmussen, David Dean (Virginia Tech, 1999-09-16)
    This study examined the efficacy of potassium lactate and lactic acid to control Campylobacter sp. and psychrotrophic bacteria on chicken. The objectives of the two studies conducted were to determine the optimal combination of potassium lactate and lactic acid to inhibit Campylobacter sp. in a challenge study and to inhibit naturally occurring Campylobacter sp. and psychrotrophic bacteria in a shelf life study. Boneless, skinless chicken breasts were injected with three levels of potassium lactate (0,1.5,2%), in conjunction with four levels of lactic acid. Lactic acid was injected (0, 0.1, 0.2, 0.3%) as well as applied directly to the surface (0.1% of weight of chicken breast). The chicken breasts were surface inoculated with a mixture of Campylobacter sp. and sampled over a period of 28 days at 11oC. The greatest inhibition was found using 2% potassium lactate in conjunction with any level of lactic acid (injected) or 0.1% lactic acid (surface application). Results of this study indicate that potassium lactate and lactic acid can be used to control the growth and/or survival of Campylobacter sp. on boneless chicken breasts. The second study eliminated the 1.5% potassium lactate and 0.2% and 0.3% lactic acid treatments and chicken breasts were not inoculated with Campylobacter sp.. This 4oC shelf life study occurred over 32 days, testing for Campylobacter species, psychrotrophic bacteria, as well as testing for sensory perceptions of color and odor changes in the chicken. The most effective treatment was the 2% potassium lactate-0.1% lactic acid surface treatment, demonstrating the most inhibition against both target populations. This treatment also had the greatest impact upon the odor of the chicken breasts. This treatment had the greatest difference from control samples, which was achieved by the inhibition of spoilage organisms on the chicken breasts.
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    Effects of Apple Development and Damage on the Internalization of Escherichia coli O157:H7 as Observed Under Field and Laboratory Conditions
    Hereford, Megan Lee (Virginia Tech, 2003-09-12)
    The number of food borne illnesses associated with the consumption of fresh fruits and vegetables and their minimally processed products (juices) has increased over the past years. Of particular interest is the ability of microbial pathogens to internalize and survive in fresh produce that are commonly used for juices. This research project addresses the issue of the ability of Escherichia coli O157:H7 to internalize and survive in whole apples before and after harvest. Four cultivars of apples, Redfree, Red Delicious, Golden Delicious, and York, were inoculated under field conditions with a surrogate strain of E. coli, Escherichia coli ATCC 25922. The Redfree cultivar was inoculated at the beginning of its growth stage (day 0), and again 30 days later, and sampled for two weeks, until E. coli was not recoverable through microbiological methods after three successive sampling days. Red Delicious, Golden Delicious, and York cultivars were spray inoculated with the surrogate strain two weeks before their anticipated harvest date and sampled every other day until E. coli was not recoverable for three successive sampling days. For each cultivar, the presence of E. coli ATCC 25922 was not detectable after 7 to 9 days. In the laboratory study the Red Delicious, Golden Delicious, Rome, and York cultivars received one of three treatments; unblemished control, bruising, or puncturing. The apples were inoculated by immersion in cold water containing E. coli O157:H7 GFP, incubated for three days then microbiologically analyzed for presence of the bacteria. In all cases, the punctured apples of each cultivar showed the greatest uptake of E. coli O157:H7 GFP. Escherichia coli O157:H7 GFP was visualized in flesh and core sections of untreated, bruised, and punctured apples of all cultivars. The microbe was found in between cells, but not within cells of the apple. Internalization of Escherichia coli in whole apples on the tree is not likely, and leads to the conclusion that internalization is a post-harvest problem. Internalization may occur before pressing or processing of apples, leading to an increased risk of infection with E. coli for consumers of apple products that are not properly treated to destroy pathogens. Internalization does occur when apples are immersed in solutions containing the pathogen Escherichia coli O157:H7, and better post harvest controls need to be implemented in order to prevent this in whole apples that are used for cider and juice production.
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    Effects of UV Irradiation on the Reduction of Bacterial Pathogens and Chemical Indicators of Milk
    Matak, Kristen E. (Virginia Tech, 2004-11-22)
    Consumer demand for fresher and minimally processed foods has brought about a movement to find effective, non-thermal processing technologies for the treatment of milk. The influence of temperature on bacterial reduction in UV irradiated milk was tested. Commercially processed skim, reduced fat (2%), and whole milk samples were inoculated with a naladixic acid resistant E. coli O157:H7 surrogate (ATCC 25922), maintained at or brought to 4oC and 20oC, respectively, and then exposed to a UV light dose between 5.3-6.3 mJ/cm2 for approximately 1.5 sec using the CiderSure 3500 apparatus (FPE Inc., Macedon, NY). Bacterial concentrations before and after UV exposure were enumerated and the results indicated that processing temperature was not significantly related to bacterial reduction (p > 0.05). The results did indicate that skim milk samples had a greater bacterial reduction, regardless of processing temperature compared to reduced fat milk and whole milk samples (p < 0.05). Solids such as milk fat, protein, lactose and minerals, in the milk have a greater effect over bacterial reductions than processing temperatures. Traditional goat cheeses are produced using unpasteurized milk, which increases the food safety concerns for these types of products. Fresh goat's milk was inoculated to 107 cfu/ml with Listeria monocytogenes (L-2289) and exposed to UV light using the CiderSure 3500 apparatus. Inoculated milk was exposed to an ultraviolet dose range between 0 and 20 mJ/cm2 to determine the optimal UV dose. A greater than 5-log reduction was achieved (p < 0.0001) when the milk was processed 12 times for a cumulative exposure time of roughly 18 sec and a cumulative UV dose of 15.8 +/- 1.6 mJ/cm2. The results of this study indicate that UV irradiation could be used for the reduction of L. monocytogenes in goat's milk. Organoleptic consequences of goat's milk treated with UV technology were assessed. Olfactory studies were conducted and a highly significant difference was determined between the odor of fresh goat's milk and UV processed milk (p < 0.05). The extent of lipid oxidation and hydrolytic rancidity was measured by thiobarbituric acid reactive substances (TBARS) and acid degree values (ADVs). Results indicated that as the UV dose increased, there was a significant increase in TBARS values and ADVs of the milk samples (p < 0.05). Milk samples were processed using the UV processor under the same conditions as previously described without exposure to the UV source to determine if the agitation from pumping was causing off-flavors by way of hydrolytic rancidity. The ADVs from these samples increased at the same rate as the UV irradiated samples; however, sensory studies indicated that the increase of free fatty acids (FFA) was not enough to cause detectable off-odors in the milk. Solid phase microextraction and gas chromatography (SPME-GC) was utilized to quantify the production of volatile compounds that were formed due to UV processing. The formation of pentanal, hexanal and heptanal was identified after as little as 1.3 mJ/cm2 UV dose. Peak areas were measured and analyzed after 7.8 mJ/cm2 and 15.6 mJ/cm2 and were determined to increase significantly as UV dose increased (p < 0.05). The chemical analyses supported the findings from the olfactory studies. The outcome of this research showed that UV irradiation at the wavelength 254 nm, was detrimental to certain chemical properties of fluid milk. The properties that were perceived as negative in fluid milk may be considered an attribute in certain types of cheese and future studies in the cheese production sector should be considered. Other applications for this technology could be for use in developing countries where milk is not typically processed because of the high costs of thermal pasteurization. On-farm applications for the treatment of replacement milk should also be considered.
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    The Efficacy of Antimicrobials for the Control of Alicyclobacillus acidoterrestris in Fruit and Vegetable Juices
    Hartman, Angela Danielle (Virginia Tech, 2003-06-12)
    The efficacy of antimicrobials for control of A. acidoterrestris spoilage in juices was analyzed. Apple and tomato juices were inoculated with 4 log spores/ml. Antimicrobials were added at: 1000, 500 and 250 ppm (sodium benzoate, potassium sorbate, and sodium metabisulfite); 500, 250, and 125 ppm (cinnamic acid, dimethyl dicarbonate, and ascorbic acid); 125, 75 and 25 ppm (lysozyme); and 5, 3, and 1 IU/ml (nisin). In apple juice, A. acidoterrestris population reductions were caused by the following antimicrobials (reduction in log CFU/ml): lysozyme - all levels and nisin - 5 IU/ml (5.1), nisin - 3 IU/ml (4.2), cinnamic acid - 125 ppm (3.1), cinnamic acid - 250 ppm (2.6), potassium sorbate - 250ppm (2.5), nisin - 1 IU/ml (2.4), potassium sorbate - 500 and 1,000 ppm (2.3), dimethyl dicarbonate - 500 ppm (1.9), cinnamic acid - 500 ppm (1.4). In tomato juice, A. acidoterrestris population reductions were caused by the following antimicrobials (reduction log CFU/ml): nisin - all levels and lysozyme - 125 ppm and 75 ppm (4.4), lysozyme - 25 ppm (3.8), potassium sorbate - 500 ppm (2.6), cinnamic acid - 500 ppm (2.5), cinnamic acid - 250 ppm (2.4), cinnamic acid - 125 ppm (2.1), potassium sorbate - 1,000 ppm (1.9), and potassium sorbate - 250 ppm (1.6). Antimicrobial treatments: nisin - ≥ 1 IU/ml, lysozyme - ≥ 25 ppm, cinnamic acid - ≥ 125 ppm, and potassium sorbate - ≥ 250 ppm may be appropriate controls to prevent A. acidoterrestris spoilage in juices or juice containing beverages.
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    Efficacy of Detergent Rinse Agents to Remove Salmonella and Shigella spp. from the Surface of Fresh Produce
    Raiden, Renee Mary (Virginia Tech, 2002-08-16)
    Fresh produce has been implicated in several foodborne outbreaks. A primary site of microbial contamination for produce occurs on the surface during production and handling. An approach to reduce contamination is to sample the surface of produce. This study used different detergent agents at 22°C and 40°C to determine their efficacy for recovery of pathogenic bacteria, from surfaces of several produce types and examined survival of organisms in detergents over time. Strawberries, tomatoes and green leaf lettuce were dip inoculated in a 6-6.5 LOG CFU/ml cocktail of nalidixic acid resistant organisms. After drying, produce were rinsed with either 0.1 % sodium lauryl sulfate (SLS), 0.1% Tween 80, or water at different temperatures. Rinse solutions were plated onto Tryptic Soy agar supplemented with 50-ppm nalidixic acid (TSAN). About 4 LOG CFU/ml of Salmonella, and 3-LOG CFU/ml Shigella were recovered, with slightly lower recovery from tomatoes. Inoculated strawberries rinsed with SLS, displayed minimal recovery at ~1.5-LOG CFU/ml at 22°C, and <1-LOG CFU/ml at 40°C. When whole strawberries treated with SLS were analyzed, few Salmonella were recovered. Lack of recovery of Salmonella rinsed with SLS, suggests SLS may be inactivating Salmonella, especially at elevated temperatures. Detergent solutions were inoculated with 3-LOG CFU/ml cocktail and incubated for up to 32 hours at 22°C, and 40°C. Aliquots were plated onto TSAN at varying times. All solutions at 40°C allowed Shigella to grow. SLS gave initial drops in Salmonella populations followed by slight recovery. SLS may cause an initial injury of Salmonella. While organisms were able to survive in detergents, the application of detergents to produce was no more effective in recovery of organisms from produce than water.
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    Efficacy of Selected Chemicals on the Attachment and Survival of Campylobacter jejuni on Chicken Breast Skin
    Arritt, Fletcher Marion (Virginia Tech, 2000-12-06)
    Campylobacter is considered to be the leading cause of acute bacterial gastroenteritis in humans in the United States with Campylobacter jejuni being responsible for 80-90% of those infections. Many cases of Campylobacter gastroenteritis have been linked to the consumption of raw or undercooked chicken. The population of bacteria on the breast skin has been reported to be greater than on other edible portions of the chicken carcass making this an important site to control the organism and to study bacterial attachment properties. This research examined the efficacy of trisodium phosphate (TSP)(10%), cetylpyridinium chloride (CPC)(0.1% & 0.5%), acidified sodium chlorite (ASC)(0.1%), Tween 80 (polysorbate 80) (1%) and water (50°C) for reducing the number of viable Campylobacter jejuni on inoculated chicken breast skin. All chemicals were evaluated using contact times of 30 sec., 3 min. or 10 min. Statistically significant (p £ 0.05) differences in the reduction of C. jejuni populations were observed across chemical treatments and contact time. When bacteria were applied before treatment, a reduction of >1.0 log10 CFU/skin was achieved with 0.5% CPC (2.89), 10% TSP (1.63), 0.1% ASC (1.52), and 0.1% CPC (1.42). When bacteria were applied after treatment, a reduction of >1.0 log10 CFU/skin was achieved with 0.5% CPC (4.67) and 10% TSP (1.28). The main effects of contact time were statistically significant (p=0.02) only when bacteria were applied after treatment.