Escherichia coli O157:H7 can contaminate dropped apples used for juicing via contact with manure or fecally tainted irrigation water and attach to the flesh of the apple through bruises and wounds where surface sanitizers are not effective. The goal of this project was to determine the efficacy of gamma irradiation at the maximum allowed dose of 1000 Gy to inactivate Escherichia coli O157: H7 in whole apples used for juicing. Whole apples were punctured to simulate wounds which were then inoculated with an outbreak strain of E.coli O157:H7 and subjected to gamma irradiation at doses upto 1000 Gy. The D-value of the E.coli O157:H7 strain was 334 Gy indicating that irradiation at 1000 Gy would result in a 3-log reduction of this pathogen. Contaminated apples were also stored for 3 weeks at refrigerated temperature during which time E.coli O157:H7 survived but did not grow. The inoculated apples were juiced, and the juice was stored up to 72 h. There was no change in counts of E.coli O157:H7 in the juice from the control apples, but irradiation at >600 Gy reduced counts by >3 logs, and survivors were not detected after 72 h storage. Sensory testing of juice treated at 652 Gy indicated consumers could tell the difference from control juice, due mostly to greater sweetness of the juice from irradiated apples. These results show that E.coli O157:H7 can easily survive in bruised apples and the juice made from them. Irradiation at 1000 Gy can provide significant lethality of E.coli O157:H7 in apples and juice conferring a greater level of safety without negative effects on sensory quality.

The extent and nature of biochemical changes that take place in canned fruits during storage temperatures above freezing have been reviewed and discussed by Pederson, et al. (1947). These changes include loss in nutritive value, e.g. ascorbic acid, thiamine (Brenner, et al., 1948) and deterioration of color (Tressler, et al., 1955). Bauernfeind (1953) reported that canned peaches, apricots, and sweet cherries, after a few months of storage at 70° F, frequently undergo changes such as destruction of anthocyanin and carotenoid pigments with the subsequent formation of brown colored compounds. Darkening of fruit-color eventually results in their unacceptability at consumer level. Preference for fruit is mainly based upon the attractive appearance of the products. Thus, color is an important factor governing the quality of fruits and fruit products. In earlier studies, conducted elsewhere, emphasis was placed on effects of low storage temperatures on the quality of canned apricots and cherries. Paucity of scientific literature on the stability of processed apricots and cherries gave impetus to a study of the comparative influence of high storage temperatures and their duration, as such tests will have considerable economic bearing upon storing and shipping processed products to tropical countries. This thesis presents the effects of storage temperatures (40, 70, 100, and 120° F) and their duration (16 weeks) on colors (anthocyanins and carotenoids), total titratable acidity, pH, viscosity, carbohydrates (total 2 and free reducing sugars, pectins), volatile reducing substances, hydroxymethyl furfural, and organoleptic quality of canned apricots and cherries .

Applications of growth regulators and modifications of light exposures were investigated regarding their abilities to curtail the normal development of chlorophyll and glycoalkaloids in potato tubers responding to light . Four growth regulators were sprayed on the foliage of potato plants 14 days prior to harvest, of which two I (Ethephon and N6 -Benzyladenine [N6BA]) successfully retarded chlorophyll and glycoalkaloids in the harvested tubers. When tubers were dipped in N6BA, lower concentrations of the !chemical promoted glycoalkaloid formation, while larger amounts we~e more inhibitory . When fluorescent lights were directed through colored cellophane filters, tubers receiving yellow and orange illuminations were inhibited in chlorophyll formation and those receiving green illumination were most reduced in production of glycoalkaloids .

Amino acids derived from the degradation of casein in cheese serve as precursors for the generation of desirable and undesirable flavor compounds. Microbial degradation of aromatic amino acids is associated with the formation of aroma compounds that impart putrid-fecal, barny-utensil, and floral off-flavors in cheese, but pathways for their production had not been established. This study investigated Tyr and Phe catabolism by Lactobacillus casei and Lactobacillus helveticus cheese flavor adjuncts under simulated Cheddar cheese-ripening conditions (pH 5.2, 4% NaCl, 15°C, no sugar). Enzyme assays of cell-free extracts and micellar electrokinetic capillary chromatography of supernatants indicated that L. casei and L. helveticus strains catabolize Tyr and Phe by successive transamination and dehydrogenation reactions. Major products of Tyr and Phe catabolism included off-flavor compounds formed by chemical degradation of the α-keto acids, produced by transamination, and aromatic α-hydroxy acids derived from α-keto acids by α-hydroxy acid dehydrogenases. Action of Lacrococcus lactis aminotransferase enzymes on Trp, Tyr, and Phe also leads to the formation of α-keto acids, but unlike lactobacilli, the former bacteria do not express dehydrogenase activity under cheese-like conditions (pH 5.2, 4% NaCl, 15°C, no sugar). Since aromatic α-keto acids may degrade spontaneously into undesirable flavor compounds, α-hydroxy acid dehydrogenases may be useful in controlling off-flavor development via diversion of chemically labile α-keto acids to more stable a-hydroxy acids. To test this hypothesis, we investigated the effect of D-hydroxyisocaproate dehydrogenase overexpression by a L. casei adjunct on chemical and sensory properties of reduced-fat Cheddar cheese made with and without addition of 20 mM α-ketoglutarate. The D-hydroxyisocaproic acid dehydrogenase gene (D-HicDH) was cloned into a high copy number vector pTRKH2 and transformed into L. casei ATCC334. Reduced-fat Cheddar cheeses were made with Lactococcus lactis starter only, starter + L. casei ATCC334 with pTRKH2, and starter + L. casei ATCC334 with pTRKH2: D-HicDH, and then volatile analysis was performed by gas chromatography and mass spectrometry. Statistical analysis of volatile data after 3 mo of ripening at 7°C showed profiles of ketones, aldehydes, alcohols, esters, sulfur compounds, and benzaldehyde were significantly altered by culture treatments and α-ketoglutarate addition, and these treatments also affected sensory flavor attributes of experimental cheeses. Results also indicated overexpression of D-hydroxyisocaproic acid dehydrogenase can divert labile α-keto acids into more stable compounds, but the overall effect seemed to diminish both beneficial and detrimental flavor notes.

High-pressure needleless injection (HPNI) is a process where small-diameter, high-velocity burst of liquid, penetrate foods at pressures ≤ 10,000 psi. The potential of HPNI as an enhancing technique for meat was studied. In study 1, HPNI translocated surface E. coli O157 into the interior of beef eye-of-round subprimals with an incidence of 40 (±7), 25 (±8), and 25 (±8)% for meat that had been surface-inoculated with a four-strain cocktail at 0.5, 1, and 2 log10 CFU/cm2, respectively. Run-off water contained 2, 2, and 3 log10 CFU/ml and was used for HPNI of additional subprimals, which resulted in a cross-contamination incidence of 83 (±4), 60 (±15), and 37 (±6) %, respectively. Incidence of translocation and cross-contamination was similar at all sampled levels below the inoculated surface. Study 1 results indicate that surface microflora will be translocated from the surface into the interior of HPNI-treated beef by the injection fluid and by cross-contamination with recycled fluid. In study 2, E. coli was undetected in cooked steaks (63˚C internal) cut from subprimals inoculated with 2 log10 CFU/cm2 and HPNI processed (study 1). Although cooking reduced E. coli counts, determination of complete kill was not possible because the detection limit for bacterial recovery was about 1 log10 CFU/g. Steaks cut from HPNI-processed subprimals took longer (p <0.05) to reach 63˚C with grilling or broiling, compared to control steaks, possibly due to increased moisture in enhanced steaks. In study 3, sensory acceptance of steaks was evaluated by a consumer panel. Appearance, flavor, and overall acceptance were similar among the untreated control, HPNI steaks, blade tenderized steaks (BT steaks), and steaks cut from subprimals that had been needle-injected with 0.35% (wt/vol) sodium tripolyphosphate using needle injection (NI-subprimal steaks) or HPNI (HPNI-subprimal steaks). Texture of BT steaks (6.5±1.9) was more liked than control steaks (5.8±1.8), while texture was similar for all other comparisons. Conversely, Warner-Bratzler shear force was NI-subprimal steaks < control < HPNI steaks = HPNI-subprimal steaks = BT steaks. Lack of correspondence between texture acceptance data and WBSF suggests that sensory scores were influenced by factors other than the force required for mechanical shear.

A whey product has been formulated for use as a lactic starter medium. Phosphate-treated whey medium (PWM) has been shown to support growth of lactic cultures and prevent phage proliferation. Comparisons were made of a commercial phage inhibitory medium (PIM), reconstituted non-fat dry milk (NDM) and PWM. PWM inhibited all phages tested and stimulated starter growth. PWM was not as stimulating as PIM but was better than NDM. Good Cheddar cheese has been made using PWM. PWM is more economical than PIM but NDM is the most economical one if cheese yield is considered.

This study sought to investigate the role of the C55 undecaprenol lipid carrier in the production of exopolysaccharide (EPS), the effect of exopolysaccharide producing (EPS+) starter cultures on the viscosity of Mozzarella cheese whey, and the possible protective characteristics of capsular EPS against freezing and freeze drying. Efforts to investigate the role of the lipid carrier in EPS production employed pAMbacA, a plasmid that encodes an enterococcallipid kinase that confers bacitracin resistance by increasing intracellular levels of undecaprenol phosphate lipid carrier. Unfortunately, this avenue of study was thwarted by the inability to demonstrate bacA expression in a model dairy lactic acid bacterium, Lactococcus lactis. To study the effect of EPS+ cultures on cheese whey, Mozzarella cheese was made with starters consisting of Lactobacillus helveticus (LH100) paired with one of four Streptococcus thermophilus strains. These strains included a capsular EPS producer (Cps+) MR-1C; a non-exopolysaccharide producing negative mutant (EPS-) of MR-1C, DM10; a ropy EPS producer, MTC360; and a non-EPS producing industrial strain, TA061. Results showed that Mozzarella cheese made with a Cps+ or ropy EPS+ S. thermophilus strain had significantly higher moisture levels than cheese made with non-exopolysaccharide producing (EPS-) streptococci. Melt properties were also better in cheeses with higher moisture. Viscosity measurements of unconcentrated and ultrafiltered (5-fold concentrated) whey showed that ultrafiltered whey from cheeses made with S. thernzophilus MTC360 was significantly higher in viscosity than whey from cheeses made with MR-1C, TA061, or DM10. There was no significant difference in the viscosity of unconcentrated or concentrated whey from cheese made with S. thermophilus MR-1C and cheese made with the commercial starter culture TA061. The results indicated that non-ropy, encapsulated exopolysaccharideproducing S. thermophilus strains can be used to achieve higher cheese moisture levels and to improve the melt properties of Mozzarella cheese without significantly increasing cheese whey viscosity. Finally, S. thermophilus MR-1C and DM10 were subjected to freezing and freeze drying to test for possible protective effects of the capsular exopolysaccharide. Analysis of variance of cell counts taken before and after freezing or freeze drying cycles revealed there was no significant difference between the viability of these strains.

One of the least controlled defects in Swiss cheese is development of splits. Split defect is characterized by fissures or cracks in the body of the cheese that can be as short as 1 cm in length or long enough to span a 90-kg block. This defect appears during refrigerated storage after the cheese is removed from the warm room. Swiss cheese with splits is downgraded because it is unsuitable for use on high-speed slicing equipment (up to 1,000 slices per minute). A 2x2x2 factorial experiment was used to determine the effect of different commercial Lactobacillus helveticus starters combined with commercial gas-forming strains of Propionibacterium freudenreichii ssp. shermanii on the occurrence of split defect in Swiss cheese. Two strains of L. helveticus recommended for Swiss cheese manufacture were used along with two strains of P. freudenreichii ssp. shermanii. The same strain of Streptococcus thermophilus was used in all treatments. To investigate the influence of seasonal variations in milk supply, eight vats were made in the summer and eight vats were made in the winter, each producing five 90-kg blocks of cheese. Each 90-kg block of cheese was cut into twenty-four 4-kg blocks, and each 4-kg block was graded based on the presence of splits. If splits were present, the cheese was downgraded from A to C grade. Only small variations were found in the composition of cheeses made during the same season. There were no correlations between cheese moisture, pH, fat, protein, calcium, lactose contents, D/L lactate ratio, or protein degradation that could be used to predict the amount of splits present after 90 d of storage. The extent of split formation was influenced by both the L. helveticus and P. freudenreichii ssp. shermanii cultures used. In this study, we were able to show a fivefold reduction in downgraded cheese through proper culture selection from 90% to 14% in the summer cheese. Even though less than 6% of the cheese split in the winter, the culture effect was nonetheless repeatable with a similar reduction through culture selection from 6% to 1% in winter cheese. Split formation also increases with storage time. If a cheese has a tendency to split, there will be a higher percentage of downgraded cheese the longer it is kept in storage.

Whole milks were adjusted to pH 5.8, 6.2, or 6. 7 with HCl and batch pasteurized at 63°C for 30 min. Each was concentrated 5:1 (40% total solids) through a single tube polysulfone membrane Abcor ultrafiltration unit. Lactose (L), casein hydrolysate (CH), and one of two brands of yeast extract (YE1, YE2) were added into cooled retentates at 0.1, 0.3, 0.5, 0. 7 or 0.9% and equilibrated overnight at 4°C. Five percent proteinase positive (Prt+) Streptococcus cremoris UC 310+ (v/w) milk based culture was added. Unfortified retentate was also inoculated with 0.1, 0.3, 0.5, 0. 7 or 0.9% starter and pH readings were taken on all samples for 24 h during incubation at 23°C. Similar substrates were inoculated with proteinase negative (Prt-) S. cremoris UC 310-. Lactose had no significant effect on acid production. Casein hydrolysate had a slight positive effect. Yeast extract had a significant effect at all preacidification levels and a significant difference was also noticed between the brands. Mean times required for the proteinase positive culture to reach pH 5.1 in 5x retentate from milk acidified to pH 5.8 were 24, 12, 10, 10, and 24 h for L, CH, YE1, YE2, and the control respectively. Proteinase negative variants of this strain had mean times of >24 h, 14 h, 11 h, 11 h, and >24 h respectively. These time differences were significantly different between Prt+ and Prt- variants. A minimum concentration of 0.2% yeast extract produced the most stimulation while greater quantities provided no additional benefit. Taste panelists were unable to detect yeast extract in retentates fermented by either culture variant.

Whey permeate was anaerobically digested in laboratory scale upflow anaerobic sludge blanket reactors. Nine hydraulic retention times between 5 and 0.2 days were examined with a fixed influent concentration of 10.6 ± 0.2 g COD/L. Chemical oxygen demand removal efficiency ranged from 99.0 to 18.9% and maximum production rate of methane gas was 2.67 L/L/day at a hydraulic loading rate of 12.97 kg COD/m3/day. About 70% of the chemical oxygen demand removed was converted to methane. Both the nonlinear least square method with 95% confidence interval and linear regression were used to evaluate kinetic coefficients. The maximum substrate utilization rate, k, and half saturation coefficient, KL, were determined to be 1.269 ± 0.163 Kg COD/kg VSS/day and 1.000 ± 0.179 kg COD/kg VSS/day. The yield coefficient, Y, and biomass decay rate coefficient, Kd, were also determined to be 0.160 ± 0.012 kg VSS/kg COD and 0.027 ± 0.004 day-1, respectively.