Association of fungal genera from spoiled processed foods with physicochemical food properties and processing conditions.

The processing conditions and physiochemical properties used in food manufacturing create niches which support the growth of a limited number of spoilage fungi. This study was designed to evaluate the influence of intrinsic and extrinsic food product variables on the identity of spoilage fungi genera isolated from commercially produced foods. The spoilage etiology was identified in 127 products through ITS region sequencing. The prevalence and diversity of the identified spoilage fungi were evaluated in relationship to product-specific attributes using various descriptive statistics and a bipartite network analysis. Additionally, recursive partitioning was used to generate a classification tree with the outcomes, genera of the spoilage isolates, divided into increasingly homogenous subgroups. All of the isolated fungi belonged to the Ascomycete phylum, except four mucoralian isolates and the basidiomycete Rhodotorula. The occurrence of filamentous fungi repeatedly isolated ranged from 2% (Phoma spp.) to 18% (Penicillium spp.). In order of decreasing contribution to subgroup homogeneity, the split rules for the classification tree were based on process, water activity, food matrix category, and pH. Fungal genera representation in the terminal nodes indicated that production failures, in addition to product-specific attributes, were responsible for determination of the most probable spoilage organism.

Isolation of Bacteriocin-producing Staphylococcus spp. Strains from Human Skin Wounds, Soft Tissue Infections and Bovine Mastitis.

A collection of 206 Staphylococcus spp. isolates was investigated for their ability to produce compounds exhibiting antistaphylococcal activity. This group included Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus xylosus strains recovered from bovine mastitis (n = 158) and human skin wounds and soft tissues infections (n = 48). Production of substances with antimicrobial activity was observed in six strains. Five of them were recovered from bovine mastitis, and one was isolated from the infected human skin wound. Three of the six antimicrobials produced by the different strains showed substantial loss of antimicrobial activity upon treatment with proteolytic enzymes, which suggests their peptidic structure. Additional studies have shown that one of the putative bacteriocins was efficiently secreted to the liquid medium, facilitating its large-scale production and isolation. The peptide produced by the M2B strain exhibited promising activity; however, against narrow spectrum of Staphylococcus spp. clinical and animal isolates. Growth inhibition was observed only in the case of 13 (including nine S. aureus, three S. xylosus and one S. epidermidis strains) out of 206 strains tested. Important advantage of the produced agent was its high thermal stability. Fifteen minutes of incubation at 90°C did not affect its antimicrobial potential. The highest efficiency of production of the agent was demonstrated in TSB medium after 24 hours at 37°C. The researches revealed that ability to production of bacteriocin among staphylococci is not very common. Only one (S. xylosus strain assigned as M2B) out of 206 strains tested produced satisfactory amounts of antistaphylococcal bacteriocin. In spite of that, we would encourage other researchers for investigation of their collections of Staphylococcus spp. isolates towards selection strains producing antimicrobial agents.

Development of a homologous expression system for and systematic site-directed mutagenesis analysis of thurincin H, a bacteriocin produced by Bacillus thuringiensis SF361.

Thurincin H is an antimicrobial peptide produced by Bacillus thuringiensis SF361. With a helical back bone, the 31 amino acids of thurincin H form a hairpin structure maintained by four pairs of very unique sulfur-to-α-carbon thioether bonds. The production of thurincin H depends on a putative gene cluster containing 10 open reading frames. The gene cluster includes three tandem structural genes (thnA1, thnA2, and thnA3) encoding three identical 40-amino-acid thurincin H prepeptides and seven other genes putatively responsible for prepeptide processing, regulation, modification, exportation, and self-immunity. A homologous thurincin H expression system was developed by transforming a thurincin H-deficient host with a novel expression vector, pGW133. The host, designated B. thuringiensis SF361 ΔthnA1 ΔthnA2 ΔthnA3, was constructed by deletion of the three tandem structural genes from the chromosome of the natural thurincin H producer. The thurincin H expression vector pGW133 was constructed by cloning the thurincin H native promoter, thnA1, and a Cry protein terminator into the Escherichia coli-B. thuringiensis shuttle vector pHT315. Thirty-three different pGW133 variants, each containing a different point mutation in the thnA1 gene, were generated and separately transformed into B. thuringiensis SF361 ΔthnA1 ΔthnA2 ΔthnA3. Those site-directed mutants contained either a single radical or conservative amino acid substitution on the thioether linkage-forming positions or a radical substitution on all other nonalanine amino acids. The bacteriocin activities of B. thuringiensis SF361 ΔthnA1 ΔthnA2 ΔthnA3 carrying different pGW133 variants against three different indicator strains were subsequently compared.

Functional assignment of YvgO, a novel set of purified and chemically characterized proteinaceous antifungal variants produced by Bacillus thuringiensis SF361.

This study reports a novel class of antifungal protein derived from bacterial origin. Bacillus thuringiensis SF361, the strain also responsible for producing the novel bacteriocin thurincin H, exhibits broad antifungal activity against select members of several fungal genera, including Aspergillus, Byssochlamys, and Penicillium, as well as the pathogenic yeast Candida albicans. Optimal antifungal production and secretion were observed after-log phase growth when incubated at 37°C in a carbohydrate-free growth medium. High-performance liquid chromatography purification was performed after pH-selective ammonium sulfate precipitation and size-exclusion chromatography. Intact mass analysis and peptide mass fingerprinting identified the 13,484-Da protein to be a mass homolog to the YvgO protein construct sequenced from Bacillus cereus AH 1134. Further analysis via amino-terminal sequencing also revealed the existence of four distinct yet equally efficacious YvgO variants differing only within the first four N-terminal residues. YvgO was found to be remarkably stable, maintaining its antifungal activity under a wide pH and temperature range. When assayed against the toxigenic species Byssochlamys fulva H25, the selected primary filamentous fungal indicator, the MIC was estimated to be 1.5 ppm. Candida albicans 3153 was more resistant, exhibiting MICs between 25 and 800 ppm, depending on growth conditions. YvgO is unique among antifungals, showing no known sequential or functional homology to the typical classes of antifungal proteins, including common membrane-acting agents such as cellulases and glucanases. Due to its activity against an array of pathogenic and spoilage fungi, the potentials for clinical, agricultural, and food-processing applications are encouraging.

Evaluation of high pressure processing (HPP) inactivation of Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes in acid and acidified juices and beverages.

Increasing consumer demand for high-quality foods has driven adoption by the food industry of non-thermal technologies such as high pressure processing (HPP). The technology is employed as a post-packaging treatment step for inactivation of vegetative microorganisms. In order to evaluate HPP inactivation of Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes in acid and acidified juices and beverages, pressure tolerance parameters were determined using log-linear and Weibull models in pH-adjusted apple juice (pH 4.5) at 5 °C. A commercial processing HPP unit was used. The Weibull model better described the inactivation kinetics of the three tested pathogens. According to estimates from the Weibull model, 1.5, 0.9, and 1.5 min are required at 600 MPa to produce 5-log reductions of E. coli, Salmonella, and L. monocytogenes, respectively, whereas according to the log-linear model, 3.2, 1.8, and 2.1 min are required. The effects of process conditions were verified using commercial products (pH between 3.02 and 4.21). In all tested commercial juices or beverages, greater than 5-log reductions were achieved for all tested pathogens using HPP process conditions of 550 MPa for 1 min. These findings demonstrate that the HPP conditions of 600 MPa for 3 min, typically used by the food industry provide an adequate safety margin for control of relevant vegetative pathogens in acid and acidified juices and beverages (pH < 4.5). Results from this study can be used by food processors to support validation studies and may be useful for the future establishment of safe harbors for the HPP industry.

Thermoaciduric Clostridium pasteurianum spoilage of shelf-stable apple juice.

Clostridium pasteurianum BB, a saccharolytic and spore-forming obligate anaerobe, was isolated and identified from shelf-stable apple juice that was responsible for multiple large spoilage outbreaks. The growth and sporulation conditions of C. pasteurianum were atypical compared with those previously published. C. pasteurianum spores were heat resistant in apple juice at pH 3.80, with D-values at 80, 85, and 90°C being 34.4, 15.9, and 4.4 min, respectively, and a z-value of 11°C. The survival curves for thermal inactivation obeyed linear first-order kinetics. Apple juice with varying pH values was used to determine the effect of pH on germination capability of C. pasteurianum spores. The spores were found to be able to germinate at pH as low as 4.3 in pH-adjusted apple juice at low contamination levels. It was confirmed by PCR that C. pasteurianum isolated from spoiled apple juice did not contain the genes for botulinum toxins B and E, which were more commonly found in neurotoxigenic butyric clostridia. Control of finished-juice pH to below 4.0 in combination with mild heating was proposed to prevent potential spoilage of shelf-stable apple juice made with spore-contaminated apple juice concentrate.

Reduction of patulin in apple cider by UV radiation.

The presence of the mycotoxin patulin in processed apple juice and cider presents a continual challenge to the food industry as both consumer health and product quality issues. Although several methods for control and/or elimination of patulin have been proposed, no unifying method has been commercially successful for reducing patulin burdens while maintaining product quality. In the present study, exposure to germicidal UV radiation was evaluated as a possible commercially viable alternative for the reduction and possible elimination of the patulin mycotoxin in fresh apple cider. UV exposure of 14.2 to 99.4 mJ/cm(2) resulted in a significant and nearly linear decrease in patulin levels while producing no quantifiable changes in the chemical composition (i.e., pH, Brix, and total acids) or organoleptic properties of the cider. For the range of UV doses tested, patulin levels decreased by 9.4 to 43.4%; the greatest reduction was achieved after less than 15 s of UV exposure. The method of UV radiation (the CiderSure 3500 system) is an easily implemented, high-throughput, and cost-effective method that offers simultaneous UV pasteurization of cider and juice products and reduction and/or elimination of patulin without unwanted alterations in the final product.

The combined effect of high pressure processing and dimethyl dicarbonate to inactivate foodborne pathogens in apple juice.

Novel processing technologies can be used to improve both the microbiological safety and quality of food products. The application of high pressure processing (HPP) in combination with dimethyl dicarbonate (DMDC) represents a promising alternative to classical thermal technologies. This research work was undertaken to investigate the combined effect of HPP and DMDC, which was aimed at reaching over 5-log reduction in the reference pathogens Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes inoculated in apple juice. Different strains of each species were tested. The pressure (ranging from 100 to 600 MPa), dwell time (from 26 to 194 s), and DMDC (from 116 to 250 mg/L) were tested based on a central composite rotatable design. The dwell time, in the studied range, did not have a significant effect (p > 0.1) on the pathogens´ reduction. All treatments achieved a greater than 5-log reduction for E. coli O157:H7 and L. monocytogenes. The reductions for S. enterica were also greater than 5-log for almost all tested combinations. The results for S. enterica suggested that it is more resistant to HPP and DMDC compared with E. coli O157:H7 and L. monocytogenes. The findings of this study showed that DMDC at low concentrations can be added to apple juice to reduce the parameters conventionally applied in HPP. The combined use of HPP and DMDC was highly effective under the conditions of this study.

Characterization of mundticin L, a class IIa anti-Listeria bacteriocin from Enterococcus mundtii CUGF08.

Enterococcus mundtii CUGF08, a lactic acid bacterium isolated from alfalfa sprouts, was found to produce mundticin L, a new class IIa bacteriocin that has a high level of inhibitory activity against the genus Listeria. The plasmid-associated operons containing genes for the mundticin L precursor, the ATP binding cassette (ABC) transporter, and immunity were cloned and sequenced. The fifth residue of the conservative consensus sequence YGNGX in the mature bacteriocin is leucine instead of valine in the sequences of the homologous molecules mundticin KS (ATO6) and enterocin CRL35. The primary structures of the ABC transporter and the immunity protein are homologous but unique.

Modeling Penicillium expansum resistance to thermal and chlorine treatments.

Apples and apple products are excellent substrates for Penicillium expansum to produce patulin. In an attempt to avoid excessive levels of patulin, limiting or reducing P. expansum contamination levels on apples designated for storage in packinghouses and/or during apple juice processing is critical. The aim of this work was (i) to determine the thermal resistance of P. expansum spores in apple juice, comparing the abilities of the Bigelow and Weibull models to describe the survival curves and (ii) to determine the inactivation of P. expansum spores in aqueous chlorine solutions at varying concentrations of chlorine solutions, comparing the abilities of the biphasic and Weibull models to fit the survival curves. The results showed that the Bigelow and Weibull models were similar for describing the heat inactivation data, because the survival curves were almost linear. In this case, the concept of D- and z-values could be used, and the D-values obtained were 10.68, 6.64, 3.32, 1.14, and 0.61 min at 50, 52, 54, 56, and 60 degrees C, respectively, while the z-value was determined to be 7.57 degrees C. For the chlorine treatments, although the biphasic model gave a slightly superior performance, the Weibull model was selected, considering the parsimony principle, because it has fewer parameters than the biphasic model has. In conclusion, the typical pasteurization regimen used for refrigerated apple juice (71 degrees C for 6 s) is capable of achieving a 6-log reduction of P. expansum spores.

Influence of storage temperature and apple variety on patulin production by Penicillium expansum.

This study examined the potential for patulin production in six different varieties of apples (Red Delicious, Golden Supreme, Gala, Fuji, Empire, and McIntosh) inoculated with Penicillium expansum spores and stored at two different temperatures (11 and 20.5 degrees C). Samples for patulin analysis were randomly taken from apples stored at different times, ranging from 21 to 93 days. While patulin was produced at both storage temperatures, apples incubated at 20.5 degrees C yielded significantly higher patulin concentrations than did those incubated at 11 degrees C. All apple varieties showed mold spoilage at both temperatures, except Red Delicious and Empire. A total of 44% of the samples analyzed showed patulin concentrations above the U.S. Food and Drug Administration regulatory limit (50 ppb). The highest patulin productions occurred in Golden Supreme (54,221 ppb) and McIntosh (52,131 and 48,457 ppb) varieties. Our results showed that careful culling of apples is essential for high juice quality, since high patulin levels in some apples varieties could result in a level greater than 50 ppb of this mycotoxin in the finished juice or cider, even when only one contaminated apple occurs in 1,000 apples.

Antimicrobial activity of bacterial isolates from different floral sources of honey.

More than two thousand bacterial strains isolated from six US domestic honeys and two manuka honeys from New Zealand were screened for production of antimicrobial compounds. A high incidence of antimicrobial inhibition determined by deferred inhibition assays was observed with the bacterial isolates from all eight honey samples. In total, 2217 isolates out of 2398 strains (92.5% of total isolates) exhibited antimicrobial activity against at least one of the tested microorganisms. Antifungal activity by bacterial isolates originating from the eight honeys ranged from 44.4% to 98.0%. Bacterial isolates from manuka honey (MH1) exhibited antimicrobial activity against Bacillus subtilis ATCC 6633 and Bacillus cereus F4552, at 51.5% and 53.3% of the isolates, respectively. However, less than 30% of the bacterial isolates from the other manuka honey (MH2) and six domestic honey sources exhibited anti-Bacillus activity. Listeria monocytogenes F2-586 1053 showed higher overall rates of sensitivity to between 11 and 66% of the bacterial isolates. The high rate of antimicrobial activity exhibited by the bacterial strains isolated from different honey sources could provide potential sources of novel antimicrobial compounds.

Efficacy of sanitizing treatments against Penicillium expansum inoculated on six varieties of apples.

The effectiveness of several wash treatments was evaluated against spores of Penicillium expansum inoculated on six varieties of apples (Red Delicious, Golden Supreme, Empire, Macintosh, Fuji, and Gala). The wash treatments were water, acidified water (pH 6.5), acidified sodium hypochlorite (pH 6.5), nonacidified sodium hypochlorite (pH 8.8, 9.3, and 9.7; 50, 100, and 200 ppm, respectively), and peracetic acid (50 and 80 ppm). Spores of P. expansum were dried on the surface of the apples for 2 h before exposure to the different sanitizer solutions. Each apple was submerged in 100 ml of each treatment solution for 30 s, and the number of spores remaining were recovered and enumerated. The efficacy of chlorine solutions was enhanced by decreasing the pH to 6.5 (up to 5-log reduction, depending on apple variety). Peracetic acid solutions (50 and 80 ppm) resulted in a reduction of less than 2 log spores per g and had the same efficacy (P < or = 0.05) as nonacidified chlorine solutions (50, 100, and 200 ppm). Control water solutions produced a reduction of 1.34 log spores per g. Chlorine solutions at pH 6.5 resulted in the largest reduction of P. expansum spores for all apple varieties tested.

Inactivation of Salmonella enterica and spoilage microorganisms in orange juice treated with dimethyl dicarbonate (DMDC).

Salmonella enterica is the pertinent pathogen associated with orange juice products that have resulted in numerous foodborne outbreaks. Although fresh orange juice typically has a pH below 4.0, which inhibits most pathogen growth, S. enterica can survive at low pH for extended periods. Additionally, fresh juice contains spoilage microorganisms such as natural yeasts and molds, which can grow at low pH, and may cause fermentation and product spoilage if left untreated. Numerous Salmonella outbreaks linked to fresh orange juice, as well as the burden of product spoilage, have generated increased demand for alternative, non-thermal treatments that can ensure pathogen- and spoilage-free products. In this study, the effect of dimethyl dicarbonate (DMDC) on pathogen and spoilage microorganism inactivation in orange juice has been investigated with two experiments. First, pasteurized orange juice was inoculated with approximately 106-107 CFU/ml of five serotypes of S. enterica per ml and treated with DMDC to test the effectiveness of inactivation against Salmonella. For the fungal spoilage microorganism study, fresh orange juice was held at room temperature to increase natural yeast and mold count to roughly 105-106 CFU/ml, followed with treatment with DMDC. DMDC at two concentrations (172 and 200 ppm) was used, and the tests were carried out at ambient (21 °C ±â€¯3 °C) and refrigeration (4 °C) temperatures. There was a >5-log reduction of Salmonella at 4 °C after 24 h at both 172 and 200 ppm of DMDC. For the treatment of fungal spoilage microorganisms, a nearly 5 and 4 log reduction of yeasts and molds was observed at ambient temperature and 4 °C, respectively. These results suggest that DMDC is most effective for use under the 4 °C holding conditions to inactivate S. enterica, and should be coupled with an additional preservative system for fungal spoilage control to produce safe orange juice that retains fresh quality.

Thermal inactivation of Salmonella and Escherichia coli O157:H7 on alfalfa seeds.

Alfalfa seeds inoculated with five strains of Salmonella or Escherichia coli O157:H7 were subjected to dry heat at 55 degrees C for up to 8 days. Five-log reductions in Salmonella or E. coli O157:H7 on seeds were observed. No pathogens were detected on the sprouted seeds, which were initially inoculated with ca. 2 log CFU/g of Salmonella or more than 8 log CFU/g of E. coli O157:H7. The percentages of germination of the alfalfa seeds did not significantly decrease after 6 days of heating at 55 degrees C. These results showed that heat treatment of alfalfa seeds at 55 degrees C for up to 6 days was effective in enhancing the safety of alfalfa sprouts without affecting germination significantly.

Effect of Water Activity on the Thermal Tolerance and Survival of Salmonella enterica Serovars Tennessee and Senftenberg in Goat's Milk Caramel.

The low thermal tolerance of Salmonella enterica in foods with intermediate moisture levels, such as caramel sauces, ensures that mild heat treatment is sufficient to achieve 5-log reductions of this pathogen. This treatment mitigates the risk posed by salmonellae in raw materials; however, recontamination might occur because of survival of the pathogen in products that are not heated before consumption. This study was conducted to evaluate the effect of water activity (aw) on the thermal tolerance and survival of S. enterica serovars Tennessee and Senftenberg. The D-values at 76, 78, and 80°C, z-values, and survival at 20.0 ± 0.5°C for 32 weeks of these two serovars were determined in goat's milk caramel at three aw values (0.85, 0.90, and 0.93). The highest thermal tolerance was observed at aw = 0.85 for Salmonella Senftenberg (D76°C = 2.9 ± 0.3 min), and the lowest was at aw = 0.93 for Salmonella Tennessee (D80°C = 0.131 ± 0.007 min). After a logarithmic transformation of the z-values, a significant interaction between serovar and aw was found (P < 0.0001), but no consistent trends were observed at the three evaluated aw levels for either serovar. Survival response was modeled using two sigmoidal three-parameter models. A significant interaction was found between nominal variables aw and serovar when comparing inflection points of the resulting curves: P < 0.0016 for the logistic model (R2 = 0.91) and P < 0.0014 for the Gompertz model (R2 = 0.92). Although a >8-log reduction was observed at week 20 of storage, regardless of the product's aw and the serovar, low levels of salmonellae were found in the product up to week 32 of storage. Our findings may assist the food industry with the establishment of critical limits for the safe thermal treatment of milk- and sugar-based foods with intermediate moisture levels. The survival data presented here highlight the relevance of implementing and effectively maintaining good sanitation and hygiene practices during the production of goat's milk caramel and similar food products.

Characterization and control of Mucor circinelloides spoilage in yogurt.

Consumer confidence in the food industry is severely affected by large-scale spoilage incidents. However, relatively little research exists on spoilage potential of members of the fungal subphylum Mucormycotina (e.g. Mucor), which includes dimorphic spoilage organisms that can switch between a yeast-like and hyphal phase depending on environmental conditions. The presence of Mucor circinelloides in yogurt may not cause spoilage, but growth and subsequent changes in quality (e.g. container bloating) can cause spoilage if not controlled. The purpose of this study was to evaluate the effects on M. circinelloides of pasteurization regimen, natamycin concentrations, and storage temperature in yogurt production, as measured by fungal proliferation and carbon dioxide production. A strain of M. circinelloides isolated from commercially spoiled yogurt showed greater yogurt-spoilage potential than clinical isolates and other industrial strains. D-values and z-values were determined for the spoilage isolate in milk as an evaluation of the fungus' ability to survive pasteurization. Natamycin was added to yogurt at 0, 5, 10, 15, and 20ppm (µg/ml) to determine its ability to inhibit M. circinelloides over the course of month-long challenge studies at 4°C, 15°C, and 25°C. Survivors were recovered on acidified PDA and carbon dioxide levels were recorded. The D-values at 54°C, 56°C, and 58°C for hyphae/sporangiospores were (in min) 38.31±0.02, 10.17±0.28, and 1.94±0.53, respectively, which yielded a z-value of 3.09°C. The D-values at 51°C, 53°C, and 55°C for yeast-like cells were (in min) 14.25±0.12, 6.87±1.19, and 2.44±0.35, respectively, which yielded a z-value of 0.34°C. These results indicated that M. circinelloides would not survive fluid milk pasteurization if contamination occurred prior to thermal treatment. CO2 production was only observed when M. circinelloides was incubated under low-oxygen conditions, and occurred only at temperatures above 4°C. Addition of 10ppm and greater of natamycin inhibited the growth and CO2 production of M. circinelloides under moderate temperature abuse when compared to the untreated control. These data suggest that yogurt spoilage (container bloating) caused by anaerobic growth of M. circinelloides is due to post-pasteurization contamination. Temperature abuse facilitated spoilage as CO2 production was observed in yogurt incubated at 15°C and 25°C, but not at 4°C. The addition of at least 10ppm of natamycin prevented M. circinelloides growth in both hyphal and yeast-like phases, as well as CO2 production in temperatures of up to 15°C for 30days.

Microbial dynamics of indicator microorganisms on fresh tomatoes in the supply chain from Mexico to the USA.

Quality and safety of fresh produce are important to public health and maintaining commerce between Mexico and USA. While preventive practices can reduce risks of contamination and are generally successful, the variable environment of the supply chain of fresh produce can be suitable for introduction or proliferation of pathogenic microorganisms. As routine surveillance of these pathogens is not practical, indicator microorganisms are used to assess the sanitary conditions of production and handling environments. An opportunity exists to use indicators on fresh produce to measure how handling and transport from field to market may affect microbial populations that contribute to their quality or safety. The objective was to quantify indicator microorganisms on tomatoes sampled along the supply chain during the harvest year, in order to observe the levels and changes of populations at different locations. Roma tomatoes (n=475) were taken from the same lots (n=28) at four locations of the postharvest supply chain over five months: at arrival to and departure from the packinghouse in México, at the distribution center in Texas, and at retail in USA. Samples were analyzed individually for four microbial populations: aerobic plate count (APC), total coliforms (TC), generic Escherichia coli, and yeasts and molds (YM). APC population differed (p<0.05) from 1.9±1.1, 1.7±1.1, 2.3±1.1 and 3.5±1.4logCFU/g at postharvest, packing, distribution center and supermarket, respectively. TC populations were <1logCFU/g at postharvest, increased at packing (0.7±1.0logCFU/g), decreased in distribution (0.4±0.8logCFU/g) and increased in supermarkets (1.4±1.5logCFU/g). Generic E. coli was not identified from coliform populations in this supply chain. YM populations remained <1logCFU/g, with the exception of 1.1±1.3logCFU/g at supermarkets and tomatoes were not visibly spoiled. The levels reported from this pilot study demonstrated the dynamics within populations as influenced by time and conditions in one supply chain during a harvest year, while the large variances in some locations indicate opportunities for improvement. Overall, packinghouse and supermarket locations were identified as crucial points to control microbial safety risks.

Variable Efficacy of the Proteinaceous Antifungal YvgO in Select Fruit Juices and Teas as a Complement with UV Methods of Food Protection.

Heat-resistant fungal spores present a processing challenge for beverages and fruit juices, as thermal and UV strategies are often inadequate in reducing heat-resistant fungal burdens to acceptable levels. While effective against pathogenic or invasive bacteria, germicidal UV light treatments also fail to achieve an appreciable reduction of heat-resistant fungal spores. As an alternative, the efficacy of the antifungal protein YvgO was examined across a selection of fruit juices and teas, as well as solid model matrices. Compared with its efficacy in analogous liquid matrices, the apparent efficacy of YvgO was diminished on acidified solid matrices due to a reduction in YvgO diffusion. Using an XTT [2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] tetrazolium dye cytotoxicity assay, the effective concentrations to reduce growth by 50% were elucidated in samples challenged with Byssochlamys fulva H25. The MICs were determined and ranged from 2 ppm in apple juice and acidified teas to approximately 3 to 12 ppm for lemonade and orange, white cranberry, blueberry, prune, cherry, and grape juices. Apple cider and nonacidified teas showed reduced efficacy, with MICs exceeding 100 ppm. Tannin-rich products readily removed YvgO from the product, impairing its efficacy. Adding bovine serum albumin as a competitive inhibitor effectively reversed the YvgO-tannin association and restored efficacy in black but not green tea matrices. When challenged with a 5-log CFU inoculum of B. fulva, the shelf lives of the products were extended for various times up to 28 days in a concentrationdependent manner. However, initial efficacy was not predictive of shelf life extension, as some products exhibited improved protection at just two- and fourfold concentrations above the MIC, while others only exhibited long-term stability when concentrations exceeded 20 times the MIC. As such, YvgO may be an attractive alternative to currently available protection strategies and will provide needed diversity for natural food protectants.

Efficient reduction of pathogenic and spoilage microorganisms from apple cider by combining microfiltration with UV treatment.

Thermal pasteurization can achieve the U. S. Food and Drug Administration-required 5-log reduction of pathogenic Escherichia coli O157:H7 and Cryptosporidium parvum in apple juice and cider, but it can also negatively affect the nutritional and organoleptic properties of the treated products. In addition, thermal pasteurization is only marginally effective against the acidophilic, thermophilic, and spore-forming bacteria Alicyclobacillus spp., which is known to cause off-flavors in juice products. In this study, the efficiency of a combined microfiltration (MF) and UV process as a nonthermal treatment for the reduction of pathogenic and nonpathogenic E. coli, C. parvum, and Alicyclobacillus acidoterrestris from apple cider was investigated. MF was used to physically remove suspended solids and microorganisms from apple cider, thus enhancing the effectiveness of UV and allowing a lower UV dose to be used. MF, with ceramic membranes (pore sizes, 0.8 and 1.4 µm), was performed at a temperature of 10 °C and a transmembrane pressure of 155 kPa. The subsequent UV treatment was conducted using at a low UV dose of 1.75 mJ/cm(2). The combined MF and UV achieved more than a 5-log reduction of E. coli, C. parvum, and A. acidoterrestris. MF with the 0.8-µm pore size performed better than the 1.4-µm pore size on removal of E. coli and A. acidoterrestris. The developed nonthermal hurdle treatment has the potential to significantly reduce pathogens, as well as spores, yeasts, molds, and protozoa in apple cider, and thus help juice processors improve the safety and quality of their products.