Yaoxin Huang

Inactivation of Escherichia coli O157:H7 and Salmonella spp. in strawberry puree by high hydrostatic pressure with/without subsequent frozen storage

The objectives of this study were to investigate the survival of Escherichia coli O157:H7 and Salmonella spp. in frozen strawberry puree and to assess the application of high pressure processing (HPP) to decontaminate strawberry puree from both pathogens. Fresh strawberry puree was inoculated with high (~6 log CFU/g) and low (~3 log CFU/g) levels of E. coli O157:H7 or Salmonella spp. and stored at -18°C for 12 weeks. Both pathogens were able to persist for at least 4weeks and samples with high inoculums were still positive for both pathogens after 12 weeks. Pressure treatment of 450 MPa for 2 min at 21°C was able to eliminate both pathogens in strawberry puree. Frozen storage at -18°C after pressure treatment substantially enhanced the inactivation of both pathogens and 4-8 days of frozen storage was able to reduce the pressure level needed for elimination of both pathogens to 250-300 MPa. Natural yeasts and molds in strawberry puree were effectively reduced by pressure of 300 MPa for 2 min at 21°C. No adverse impacts on physical properties such as color, soluble solids content, pH and viscosity of strawberry puree was found for pressure-treated samples. Therefore, the treatment of 300 MPa for 2 min at 21°C followed by 4 days frozen storage at -18°C was recommended for the minimal processing of strawberry puree with great retention of fresh-like sensory properties. HPP could be a promising alternative to traditional thermal processing for berry purees.

Inactivation of Vibrio parahaemolyticus and Vibrio vulnificus in oysters by high-hydrostatic pressure and mild heat.

Several recent outbreaks associated with oysters have heightened safety concerns of raw shellfish consumptions, with the majority being attributed to Vibrio spp. The objective of this study was to determine the effect of high-hydrostatic pressure (HHP) followed by mild heating on the inactivation of Vibrio parahaemolyticus and Vibrio vulnificus in live oysters. Inoculated oysters were randomly subjected to: a) pressurization at 200-300 MPa for 2 min at 21 °C, b) mild heat treatment at 40, 45 or 50 °C for up to 20 min and c) pressure treatment of 200-300 MPa for 2 min at 21 °C followed by heat treatment at 40-50 °C. Counts of V. parahaemolyticus and V. vulnificus were then determined using the most probable number (MPN) method. Pressurization at 200-300 MPa for 2 min resulted in various degrees of inactivation, from 1.2 to >7 log MPN/g reductions. Heat treatment at 40 and 45 °C for 20 min only reduced V. parahaemolyticus and V. vulnificus by 0.7-2.5 log MPN/g while at 50 °C for 15 min achieved >7 log MPN/g reduction. HHP and mild heat had synergistic effects. Combinations such as HHP at 250 MPa for 2 min followed by heat treatment at 45 °C for 15 min and HHP at 200 MPa for 2 min followed by heat treatment at 50 °C for 5 min reduced both V. parahaemolyticus and V. vulnificus to non-detectable levels by the MPN method (<3 MPN/g). HHP at ≥275 MPa for 2 min followed by heat treatment at 45 °C for 20 min and HHP at ≥200 MPa for 2 min followed by heat treatment at 50 °C for 15 min completely eliminated both pathogens in oysters (negative enrichment results). This study demonstrated the efficiency of HHP followed by mild heat treatments on inactivation of V. parahaemolyticus and V. vulnificus and could help the industry to establish parameters for processing oysters.

Efficacy of washing with hydrogen peroxide followed by aerosolized antimicrobials as a novel sanitizing process to inactivate Escherichia coli O157:H7 on baby spinach.

Aerosolization was investigated as a potential way to apply allyl isothiocyanate (AIT), hydrogen peroxide (H(2)O(2)), acetic acid (AA) and lactic acid (LA) on fresh baby spinach to control Escherichia coli O157:H7 during refrigeration storage. In this study, baby spinach leaves were dip-inoculated with E. coli O157:H7 to a level of 6 log CFU/g and stored at 4°C for 24 h before treatment. Antimicrobials were atomized into fog-like micro-particles by an ultrasonic nebulizer and routed into a jar and a scale-up model system where samples were treated. Samples were stored at 4°C for up to 10 days before the survival of the cells was determined. A 2-min treatment with 5% AIT resulted in a >5-log reduction of E. coli O157:H7 on spinach after 2 days refrigeration regardless if the samples were pre-washed or not; however, this treatment impaired the sensory quality of leaves. Addition of LA to AIT improved the antimicrobial efficacy of AIT. In the jar system, washing with 3% H(2)O(2) followed by a 2-min treatment of 2.5% LA+1% AIT or 2.5% LA+2% AIT reduced E. coli O157:H7 population by 4.7 and >5 log CFU/g, respectively, after 10 days refrigeration. In the scale-up system, up to 4-log reduction of bacterial population was achieved for the same treatments without causing noticeable adverse effect on the appearance of leaves. Thus, this study demonstrates the potential of aerosolized AIT+LA as a new post-washing intervention strategy to control E. coli O157:H7 on baby spinach during refrigeration storage.

A novel water-assisted pulsed light processing for decontamination of blueberries.

Sample heating and shadowing effect have limited the application of pulsed light (PL) technology for decontamination of fresh produce. In this study, a novel setup using water-assisted PL processing was developed to overcome these limitations. Blueberries inoculated with Escherichia coli O157:H7 or Salmonella were either treated with PL directly (dry PL treatment) or immersed in agitated water during the PL treatment (wet PL treatment) for 5-60 s. Although both pathogens were effectively inactivated by the dry PL treatments, the appearance of the blueberries was adversely affected and a maximum temperature of 64.8 °C on the blueberry surface was recorded. On the other hand, the visual appearance of blueberries remained unchanged after wet PL treatments and sample heating was significantly reduced. The wet PL treatments were more effective than chlorine washing on inactivating both pathogens. After a 60-s wet PL treatment, the populations of E. coli O157:H7 inoculated on calyx and skin of blueberries were reduced by 3.0 and >5.8 log CFU/g, respectively. Salmonella on blueberry calyx and skin was reduced by 3.6 and >5.9 log CFU/g, respectively. No viable bacterial cells were recovered from the water used in the wet PL treatments, demonstrating that this setup could prevent the risk of cross-contamination during fresh produce washing. Our results suggest that this new water-assisted PL treatment could be a potential non-chemical alternative (residue free) to chlorine washing since it is both more effective and environmentally friendly than chlorine washing.

Effects of pre- or post-processing storage conditions on high-hydrostatic pressure inactivation of Vibrio parahaemolyticus and V. vulnificus in oysters.

The effects of storage conditions on subsequent high-hydrostatic pressure (HHP) inactivation of Vibrio parahaemolyticus and Vibrio vulnificus in oysters were investigated. Live oysters were inoculated with V. parahaemolyticus or V. vulnificus to ca. 7-8 log MPN/g by feeding and stored at varying conditions (i.e., 21 or 35 °C for 5h, 4 or 10 °C for 1 and 2 days and -18 °C for 2 weeks). Oyster meats were then treated at 225-300 MPa for 2 min at 4, 21 or 35 °C. HHP at 300 MPa for 2 min achieved a >5-log MPN/g reduction of V. parahaemolyticus, completely inactivating V. vulnificus (negative by enrichment) in oysters. Treatment temperatures of 4, 21 and 35 °C did not significantly affect pressure inactivation of V. parahaemolyticus or V. vulnificus (P>0.05). Cold storage at -18, 4 and 10 °C, prior to HHP, decreased V. parahaemolyticus or V. vulnificus populations by 1.5-3.0 log MPN/g, but did not increase their sensitivity to subsequent HHP treatments. The effects of cold storage after HHP on inactivation of V. parahaemolyticus in oysters were also determined. Oysters were inoculated with V. parahaemolyticus and stored at 21 °C for 5h or 4 °C for 1 day. Oyster meats were then treated at 250-300 MPa for 2 min at 21 or 35 °C and stored for 15 days in ice or in a freezer. V. parahaemolyticus populations in HHP-treated oysters gradually decreased during post-HHP ice or frozen storage. A validation study using whole-shell oysters was conducted to determine whether the presence of oyster shells influenced HHP inactivation of V. parahaemolyticus. No appreciable differences in inactivation between shucked oyster meat and whole-shell oysters were observed. HPP at 300 MPa for 2 min at 21 °C, followed by 5-day ice storage or 7-day frozen storage, and HPP at 250 MPa for 2 min at 21 °C, followed by 10-day ice or 7-day frozen storage, completely inactivated V. parahaemolyticus in whole-shell oysters (>7 log reductions). The combination of HHP at a relatively low pressure (e.g., 250 MPa) followed by short-term frozen storage (7 days) could potentially be applied by the shellfish industry as a post-harvest process to eliminate V. parahaemolyticus in oysters.

Decontamination of Escherichia coli O157:H7 on green onions using pulsed light (PL) and PL–surfactant–sanitizer combinations

Imported green onion has been associated with three large outbreaks in the USA. Contamination has been found on both domestic and imported green onions. The objective of our study was to investigate Escherichia coli O157:H7 inactivation efficacy of pulsed light (PL) as well as its combination with surfactant and/or sanitizers on green onions. Green onions were cut into two segments, stems and leaves, to represent two different matrixes. Stems were more difficult to be decontaminated. Spot and dip inoculation methods were compared and dipped inoculated green onions were found to be more difficult to be decontaminated. Results showed that 5s dry PL (samples were not immersed in water during PL treatment) and 60s wet PL (samples were immersed in water and stirred during PL treatment) treatments provided promising inactivation efficacy (>4log10CFU/g) for spot inoculated stems and leaves. For dip inoculated green onions, 60s wet PL treatment was comparable with 100ppm chlorine washing, demonstrating that PL could be used as an alternative to chlorine. To further increase the degree of microbial inactivation, combined treatments were applied. PL combined with surfactant (SDS) was found to be more effective than single treatments of PL, SDS, chlorine, citric acid, thymol, and hydrogen peroxide, and binary combined treatments of PL with one of those chemicals. Addition of chlorine or hydrogen peroxide to the PL-SDS combination did not further enhanced its microbial inactivation efficacy. The combination of PL and 1000ppm of SDS reduced the E. coli O157:H7 populations dip inoculated on the stems and leaves of green onions by 1.4 and 3.1log10CFU/g, respectively. Our findings suggest that PL could potentially be used for decontamination of E. coli O157:H7 on green onions, with wet PL added with SDS being the most effective PL treatment.

Application of water-assisted pulsed light treatment to decontaminate raspberries and blueberries from Salmonella

We developed and evaluated a small scaled-up water-assisted pulsed light (WPL) system, in which berries were washed in a flume washer while being irradiated by pulsed light (PL). Hydrogen peroxide (H2O2) was used in combination with PL as an advanced oxidation process and chlorine wash was used as a control. The effects of organic load, water turbidity, berry type and PL energy output on the inactivation of Salmonella using the WPL system were investigated. The combination of WPL and 1% H2O2 (WPL-H2O2) was the most effective treatment which reduced Salmonella on raspberries and blueberries by 4.0 and >5.6logCFU/g, respectively, in clear water. When high organic load and SiO2, as a soil simulator, were added in wash water, the free chlorine level in chlorinated water decreased significantly (P<0.05); however, no significant difference (P>0.05) was observed for the decontamination efficacy of 1-min WPL-H2O2 treatment. Even in the presence of high organic load and water turbidity, no viable bacterial cells were recovered from the wash water, which showed that WPL-H2O2 could effectively prevent the risk of cross-contamination during treatment. Taken together, 1-min WPL treatment without H2O2 could provide a chemical free alternative to chlorine washing with similar and in some cases significantly higher bactericidal efficacy. Compared with chlorine washing, the combination of WPL and H2O2 resulted in significantly higher (P<0.05) reduction of Salmonella on berries, providing a novel intervention for processing of small berries intended for fresh-cut and frozen berry products.

Strategies to enhance high pressure inactivation of murine norovirus in strawberry puree and on strawberries.

Due to the increasing concern of viral infection related to berries, this study investigated strategies to enhance high hydrostatic pressure (HHP) inactivation of murine norovirus 1 (MNV-1), a human norovirus (HuNoV) surrogate, on strawberries and in strawberry puree. Strawberry puree was inoculated with ~10(6)PFU/g of MNV-1 and treated at 350 MPa for 2 min at initial sample temperatures of 0, 5, 10 and 20°C. MNV-1 became more sensitive to HHP as initial sample temperature decreased from 20 to 0°C. To determine the effect of pressure cycling on MNV-1 inactivation, inoculated puree samples were treated at 300 MPa and 0°C with 1, 2 and 4 cycles. Pressure cycling offered no distinct advantage over continuous HHP treatment. To determine the effect of presence of water during HHP on MNV-1 inactivation, strawberries inoculated with ~ 4 × 10(5)PFU/g of MNV-1 were either pressure-treated directly (dry state) or immersed in water during pressure treatment. MNV-1 was very resistant to pressure under the dry state condition, but became sensitive to pressure under the wet state condition. The inactivation curves of MNV-1 in strawberry puree and on strawberries were obtained at 300 and 350 MPa and initial sample temperature of 0°C. Except for the curve of strawberries treated at 350 MPa which had a concave downward shape, the other three curves were almost linear with R(2) value of 0.99. The fate of MNV-1 in the un-treated and pressure-treated strawberries and strawberry puree during frozen storage was determined. The virus was relatively stable and only reduced by <1.2 log during the 28-day frozen storage. In all, this study provides practical insights of designing strategies using HHP to inactivate HuNoV on strawberries and in strawberry puree assuming that HuNoV behaved similarly to MNV-1 when treated by HHP.

Inactivation of Escherichia Coli O157:H7 and Salmonella Enterica on Blueberries in Water Using Ultraviolet Light

UNLABELLED Ultraviolet light (UV) has antimicrobial effects, but the shadowing effect has limited its application. In this study, a novel setup using UV processing in agitated water was developed to inactivate Escherichia coli O157:H7 and Salmonella on blueberries. Blueberries were dip- or spot-inoculated with E. coli or Salmonella. Blueberries inoculated with E. coli were treated for 2 to 10 min with UV directly (dry UV) or immersed in agitated water during UV treatment (wet UV). E. coli was most easily killed on spot-inoculated blueberries with a 5.2-log reduction after 10-min wet UV treatment. Dip-inoculated blueberries were the most difficult to be decontaminated with only 1.6-log reduction after 10-min wet UV treatment. Wet UV treatment generally showed higher efficacies than dry UV treatment, achieving an average of 1.4 log more reduction for spot-inoculated blueberries. For dip-inoculated blueberries, chlorine washing and UV treatments were less effective, achieving <2 log reductions of E. coli. Thus, the efficacy of combinations of wet UV with sodium dodecyl sulfate (SDS), levulinic acid, or chlorine was evaluated. Inoculated blueberries were UV-treated while being immersed in agitated water containing 100 ppm SDS, 0.5% levulinic acid or 10 ppm chlorine. The 3 chemicals did not significantly enhance the wet UV treatment. Findings of this study suggest that UV treatment could be used as an alternative to chlorine washing for blueberries and potentially for other fresh produce. PRACTICAL APPLICATION A novel UV light system for decontamination of blueberries in water was developed and evaluated. Results demonstrated that the decontamination efficacy of this system was generally as effective as chlorine washing, indicating that it could potentially be used as an alternative to chlorine washing for blueberries and other fresh produce.

Effects of High-Hydrostatic Pressure on Inactivation of Human Norovirus and Physical and Sensory Characteristics of Oysters.

The purpose of the study was to determine the effect of high-hydrostatic pressure (HHP) on inactivation of human norovirus (HuNoV) in oysters and to evaluate organoleptic characteristics of oysters treated at pressure levels required for HuNoV inactivation. Genogroup I.1 (GI.1) or Genogroup II.4 (GII.4) HuNoV was inoculated into oysters and treated at 300 to 600 MPa at 25 and 0 °C for 2 min. After HHP, viral particles were extracted by porcine gastric mucin-conjugated magnetic beads (PGM-MBs) and viral RNA was quantified by real-time RT-PCR. Lower initial temperature (0 °C) significantly enhanced HHP inactivation of HuNoV compared to ambient temperature (25 °C; P < 0.05). HHP at 350 and 500 MPa at 0 °C could achieve more than 4 log10 reduction of GII.4 and GI.1 HuNoV in oysters, respectively. HHP treatments did not significantly change color or texture of oyster tissue. A 1- to 5-scale hedonic sensory evaluation on appearance, aroma, color, and overall acceptability showed that pressure-treated oysters received significantly higher quality scores than the untreated control (P < 0.05). Elevated pressure levels at 450 and 500 MPa did not significantly affect scores compared to 300 MPa at 0 °C, indicating increasing pressure level did not affect sensory acceptability of oysters. Oysters treated at 0 °C had slightly lower acceptability than the group treated at room temperature on day 1 (P < 0.05), but after 1 wk storage, no significant difference in sensory attributes and consumer desirability was observed (P > 0.05).