Inactivation of pathogens on pork by steam-ultrasound treatment

The objective of the study was to evaluate a new pathogen inactivation concept that combines application of pressurized steam simultaneously with high-power ultrasound through a series of nozzles. On skin and meat surfaces of pork jowl samples, counts of total viable bacteria were reduced by 1.1 log CFU/cm(2) after treatment for 1 s and by 3.3 log CFU/cm(2) after treatment for 4 s. The mean reduction of 1.7 to 3.3 log CFU/cm(2) on the skin surface was significantly higher than the reduction of 1.1 to 2.5 log CFU/cm(2) on the meat surface. The inactivation of Salmonella Typhimurium, Salmonella Derby, Salmonella Infantis, Yersinia enterocolitica, and a nonpathogenic Escherichia coli was studied on inoculated samples that were treated for 0.5 to 2.0 s. With one exception, no significant differences in reduction were observed among the bacterial types. After treatment for 0.5 s, the 0.9-to 1.5-log reductions of E. coli were significantly higher than the 0.4- to 1.1-log reductions for Salmonella and Y. enterocolitica. Overall, reductions increased by increasing treatment time; reductions were 0.4 to 1.5 log CFU/cm(2) after treatment for 0.5 s and 2.0 to 3.6 log CFU/cm(2) after treatment for 2 s. Reductions on the skin (1 to 3.6 log CFU/cm(2)) were significantly higher than reductions on the meat surface (1 to 2.5 log CFU/cm(2)). The reduced effect on the meat surface may be explained by greater protection of bacteria in deep structures at the muscle surface. No significant difference in reduction was observed between samples inoculated with 10(4) CFU/cm(2) and those inoculated with 10(7) CFU/cm(2), and cold storage of samples for 24 h at 5°C after steam-ultrasound treatment did not lead to changes in recovery of bacteria.     

Inactivation of Escherichia coli O157:H7, Salmonella and human norovirus surrogate on artificially contaminated strawberries and raspberries by water-assisted pulsed light treatment

This study investigated the inactivation of Escherichia coli O157:H7, Salmonella and murine norovirus (MNV-1), a human norovirus surrogate, on strawberries and raspberries using a water-assisted pulsed light (WPL) treatment. The effects of combinations of WPL treatment with 1% hydrogen peroxide (H₂O₂) or 100 ppm sodium dodecyl sulfate (SDS) were also evaluated. Strawberries and raspberries were inoculated with E. coli O157:H7 and treated by WPL for 5–60 s. E. coli O157:H7 on both strawberries and raspberries was significantly reduced in a time-dependent manner with 60-s WPL treatments reducing E. coli O157:H7 by 2.4 and 4.5 log CFU/g, respectively. Significantly higher reductions of E. coli O157:H7 were obtained using 60-s WPL treatment than washing with 10 ppm chlorine. Compared with washing with chlorine, SDS and H₂O₂, the combination of WPL treatment with 1% H₂O₂ for 60 s showed significantly higher efficacy by reducing E. coli O157:H7 on strawberries and raspberries by 3.3- and 5.3-log units, respectively. Similarly, Salmonella on strawberries and raspberries was inactivated by 2.8- and 4.9-log units after 60-s WPL–H₂O₂ treatments. For decontamination of MNV-1, a 60-s WPL treatment reduced the viral titers on strawberries and raspberries by 1.8- and 3.6-log units, respectively and the combination of WPL and H₂O₂ did not enhance the treatment efficiency. These results demonstrated that the WPL treatment can be a promising chemical-free alternative to chlorine washing for decontamination of berries destined for fresh-cut and frozen berry products. WPL–H₂O₂ treatment was the most effective treatment in our study for decontamination of bacterial pathogens on berries, providing an enhanced degree of microbiological safety for berries.     

Ozone inactivation of norovirus surrogates on fresh produce

Preharvest contamination of produce by foodborne viruses can occur through a variety of agents, including animal feces/manures, soil, irrigation water, animals, and human handling. Problems of contamination are magnified by potential countrywide distribution. Postharvest processing of produce can involve spraying, washing, or immersion into water with disinfectants; however, disinfectants, including chlorine, have varying effects on viruses and harmful by-products pose a concern. The use of ozone as a disinfectant in produce washes has shown great promise for bacterial pathogens, but limited research exists on its efficacy on viruses. This study compares ozone inactivation of human norovirus surrogates (feline calicivirus [FCV] and murine norovirus [MNV]) on produce (green onions and lettuce) and in sterile water. Green onions and lettuce inoculated with FCV or MNV were treated with ozone (6.25 ppm) for 0.5- to 10-min time intervals. Infectivity was determined by 50% tissue culture infectious dose (TCID(50)) and plaque assay for FCV and MNV, respectively. After 5 min of ozone treatment, >6 log TCID(50)/ml of FCV was inactivated in water and ～2-log TCID(50)/ml on lettuce and green onions. MNV inoculated onto green onions and lettuce showed a >2-log reduction after 1 min of ozone treatment. The food matrix played the largest role in protection against ozone inactivation. These results indicate that ozone is an alternative method to reduce viral contamination on the surface of fresh produce.     

Chlorine treatment to inactivate norovirus on food contact surfaces

This study was conducted to determine the concentration and optimal treatment time of chlorine for reducing feline calicivirus (FCV) and murine norovirus (MNV) as surrogates of norovirus (NoV) on stainless steel surfaces and to develop a predictive inactivation method using a response surface methodology. The reduction levels of FCV VR-782 and MNV on stainless steel surfaces after treatment with various concentrations of chlorine (0 to 5,000 ppm) for various times (0 to 5 min) were measured. The reduction values of both FCV and MNV on stainless steel surfaces after 5,000 ppm of chlorine treatment for 5 min were 5.20 TCID(50) per coupon. The predictive results obtained by central composite design were analyzed by standard analysis of variance. The application of multiple regression analysis was related to the following polynomial equations: (i) FCV (log TCID(50) per coupon) = -0.3714 + 0.8362x(1) + 0.0011x(2) + 0.0001x(1)x(2) - 0.1143x(2)(1) -0.0001x(2)(2) (x(1), time; x(2), concentration) and (ii) MNV (log TCID(50) per coupon) = + 0.0471 + 0.0807x(1) + 0.0011x(2) + 0.0001x(1)x(2) -0.0910x(2)(1) -0.0001x(2)(2) (x(1), time; x(2), concentration). It was concluded that these polynomial equation models of reduction of FCV and MNV could be used to determine the minimum concentration of chlorine and exposure times to control human NoV on food contact surfaces.     

Preservation treatment of fresh raspberries by e-beam irradiation

E-beam irradiation was studied as a post-harvest treatment for red raspberries (Rubus idaeus L.). Microbial inactivation (natural microbiota and potential pathogenic bacteria) and bioactive properties (phenolic content, vitamin C content and antioxidant activity and cytotoxicity) of these fruits were evaluated before and after irradiation and during storage of 14 days at 4 °C. A reduction of 2 log CFU/g of mesophilic bacteria and 3 log CFU/g on filamentous fungi, and no detection of foodborne inoculated pathogens (3 log CFU/g) was achieved with an e-beam treatment at 3 kGy and during 7 days of refrigerated storage. Regarding bioactive properties, the results suggested that irradiation could preserve the phenolic content and antioxidant activity of raspberries through 7 days of cold storage, even though a decrease of 80% on ascorbic acid concentration was observed. Furthermore, no in vitro inhibitory effect on human cells lines was observed for the extracts from e-beam-treated raspberries. The overall results suggested that use of e-beam irradiation as post-harvest treatment of raspberries as an emergent, clean and environmental friendly process to extend the shelf-life of this fruit with safety and preservation of bioactivity.Industrial relevanceRed raspberries are known to demonstrate high bioactivity that could be beneficial to human health, but are highly perishable and often associated with foodborne outbreaks, which makes its safety and commercialization a challenge. The use of a terminal control such as irradiation might reduce the burden of disease transmission and extend the quality of fresh red raspberries. The present research indicated that e-beam irradiation can be used as post-harvest treatment of raspberries, guarantying its safety and quality with the add-value of shelf-life extension.     

Differences in the binding of human norovirus to and from romaine lettuce and raspberries by water and electrolyzed waters

Food contamination by human norovirus (hNoV) is a major cause of gastrointestinal disease. We evaluated the effectiveness of removing inoculated hNoV from the surfaces of raspberries and romaine lettuce by a simple wash in tap water and in different forms of electrolyzed water (EW), including acidic EW (AEW), neutral EW (NEW), and basic EW (BEW). A simple rinsing or soaking in water was able to remove >95% of hNoV from surface-inoculated raspberries. In contrast, only 75% of hNoV was removed from surface-inoculated romaine lettuce by rinsing in tap water. An AEW wash enhanced the binding of hNoV to raspberries and lettuce. Only 7.5% (±10%) and 4% (±3.1%) of hNoV were removed by AEW wash from surface-inoculated raspberries and lettuce, respectively. When raspberries and lettuce were prewashed with NEW or BEW prior to surface inoculation, an AEW wash likewise resulted in significantly less removal of hNoV compared with untreated samples. A prewash with AEW significantly decreased the removal of hNoV from raspberries and lettuce when they were washed with NEW, from 90.6 to 51% and from 76 to 51.3% , respectively. There are minimal or no improvements gained by use of any of the EWs instead of a regular tap water wash in removal of hNoV from produce. However, use of AEW shows a significant decrease in the removal of hNoV from contaminated produce compared with other water rinses. The ability to remove hNoV from different types of produce varies, possibly due to differences among types of ligand-like molecules that bind hNoV. The distribution of hNoV on raspberries and lettuce was studied using recombinant Norwalk-like particles (rNVLP). By immunofluorescence microscopy, we were able to observe binding of rNVLP only to vein areas of romaine lettuce, suggesting that the virus was binding to specific molecules in these areas. Random binding of rNVLP occurred only with raspberries prewashed with AEW or washed with AEW.     

Inactivation of feline calicivirus and murine norovirus during Dongchimi fermentation

Among the traditional fermented vegetables in Korea, Dongchimi is a type of kimchi with a large water base. We aimed to investigate the survival of norovirus surrogates during Dongchimi fermentation. Dongchimi spiked with feline calicivirus (FCV) or murine norovirus (MNV) was prepared following a traditional recipe. Dongchimi was initially fermented at room temperature overnight and then kept at 4 °C. The number of lactic acid bacteria, pH, acidity, and virus titer were measured 0, 1, 2, 3, 5, 7, 10, 15, 20 days after fermentation. During the fermentation process, lactic acid bacteria and acidity increased. At the end of the fermentation, population of FCV and MNV decreased about 4.12 and 1.47 log units, respectively. Based on the significant reduction of norovirus surrogate during Dongchimi fermentation, we conclude that the risk of norovirus in Dongchimi may be low.     

Inactivation of feline calicivirus as a surrogate for norovirus on lettuce by electron beam irradiation

Caliciviridae, including norovirus, are considered important sources of human gastroenteritis. As leafy green vegetables are commonly consumed without additional processing, it is important to evaluate interventions to reduce the presence of human pathogens in these products. Feline calicivirus was used as a model for small round structured viruses on lettuce. The lettuce was inoculated by immersion to simulate contamination from irrigation or wash water. The inoculated lettuce was then exposed to electron beam irradiation at various dose levels to determine survival. The D??-value of the calicivirus on lettuce was determined to be 2.95 kGy. Irradiation to reduce bacterial pathogens on cut lettuce could also reduce the risk associated with small round structured viruses on lettuce.     

Inactivation of hepatitis A virus,poliovirus and a norovirus surrogate by high pressure processing

Hepatitis A virus (HAV), feline calicivirus (FCV, a surrogate for non-culturable norovirus), and poliovirus (PV), suspended in buffered cell culture media, were treated with pressures ranging from 200 to 600 MPa at ambient temperature for between 30 and 600 s. HAV was inactivated by > 1-log₁₀ tissue culture infectious dose 50% mL^− 1 (TCID₅₀ mL^− 1) and > 2-log₁₀ TCID₅₀ mL^− 1 after 600 s treatment with 300 and 400 MPa, respectively, and was undetectable (> 3.5-log₁₀ TCID₅₀ mL^− 1 reduction) within 300 s treatment with 500 MPa. FCV was inactivated by 3.6-log₁₀ TCID₅₀ mL^− 1 after 120 s treatment with 300 MPa, and was undetectable after 180 s treatment with 300 MPa. PV was the most resistant with little or no substantial reduction in titre after 300 s treatment with 600 MPa. The studies were designed to determine the efficacy of using high pressure to inactivate enteric viruses and generate inactivation data to assist in determining appropriate process criteria for safe shellfish production.Industrial relevanceThe high pressure treatment of raw oysters has proved commercially successful, due in part to a marked increase in the product’s shelf life, yet little alteration of its organoleptic properties. Illnesses from human enteric viruses such as hepatitis A virus and norovirus have traditionally been associated with shellfish consumption, and for this reason, studies have examined the stability of enteric viruses under high pressure. However, kinetic data on enteric virus stability under pressure is needed by processors to better understand the response of viruses throughout the entire treatment time. The kinetic data obtained in this study may be useful for processors wishing to alter high pressure processing conditions to ensure a high quality product in terms of organoleptic and microbiological properties.     

Efficacy of a levulinic Acid plus sodium dodecyl sulfate-based sanitizer on inactivation of human norovirus surrogates

Human noroviruses are the most common etiologic agent of foodborne illness in the United States. The inability to culture human noroviruses in the laboratory necessitates the use of surrogate viruses such as murine norovirus (MNV-1) and feline calicivirus (FCV) for inactivation studies. In this study, a novel sanitizer of organic acid (levulinic acid) plus the anionic detergent sodium dodecyl sulfate (SDS) was evaluated. Viruses were treated with levulinic acid (0.5 to 5%), SDS (0.05 to 2%), or combinations of levulinic acid plus SDS (1:10 solution of virus to sanitizer). MNV-1 inoculated onto stainless steel also was treated with a 5% levulinic acid plus 2% SDS liquid or foaming solution. Log reductions of viruses were determined with a plaque assay. Neither levulinic acid nor SDS alone were capable of inactivating MNV-1 or FCV, resulting in a ≤0.51-log reduction of the infectious virus titer. However, the combination of 0.5% levulinic acid plus 0.5% SDS inactivated both surrogates by 3 to 4.21 log PFU/ml after 1 min of exposure. Similarly, MNV-1 inoculated onto stainless steel was reduced by >1.50 log PFU/ml after 1 min and by >3.3 log PFU/ml after 5 min of exposure to a liquid or foaming solution of 5% levulinic acid plus 2% SDS. The presence of organic matter (up to 10%) in the virus inoculum did not significantly affect sanitizer efficacy. The fact that both of the active sanitizer ingredients are generally recognized as safe to use as food additives by the U.S. Food and Drug Administration further extends its potential in mitigating foodborne disease.     

Inactivation of hepatitis A virus and norovirus surrogate in suspension and on food-contact surfaces using pulsed UV light (pulsed light inactivation of food-borne viruses)

This study was conducted to evaluate the inactivation of murine norovirus (MNV-1) and hepatitis A virus (HAV) by pulsed ultraviolet (UV) light. MNV-1 was used as a model for human norovirus. Viral suspensions of about 10⁶ PFU/ml were exposed to pulses of UV light for different times and at different distances in a Xenon Steripulse device (model RS-3000C). Inactivation studies were also carried out on 1-cm² stainless steel and polyvinyl chloride disks with 10⁵ PFU/ml. Inactivation of MNV-1 and HAV at 10.5 cm from the UV source was greater on inert surfaces than in suspension. The presence of organic matter (fetal bovine serum) reduced the effectiveness of pulsed light both in suspension and on surfaces. However, 2-s treatment in the absence of FBS completely inactivated (5 log reduction) the viral load at different distances tested, whether in suspension (MNV-1) or on disks (MNV-1 and HAV). The same treatment in the presence of fetal bovine serum (5%) allowed a reduction of about 3 log. This study showed that short duration pulses represent an excellent alternative for inactivation of food-borne viruses. This technology could be used to inactivate viruses in drinking water or on food-handling surfaces.     

Inactivation of Listeria monocytogenes on agar and processed meat surfaces by atmospheric pressure plasma jets

An apparatus for generating atmospheric pressure plasma (APP) jet was used to investigate the inactivation of Listeria monocytogenes on the surface of agar plates and slices of cooked chicken breast and ham. He, N₂ (both 7 L/min), and mixtures of each with O₂ (0.07 L/min) were used to produce the plasma jets. After treatment for 2 min with APP jets of He, He + O₂, N₂, or N₂ + O₂, the numbers of L. monocytogenes on agar plates were reduced by 0.87, 4.19, 4.26, and 7.59 log units, respectively. Similar treatments reduced the L. monocytogenes inoculated onto sliced chicken breast and ham by 1.37 to 4.73 and 1.94 to 6.52 log units, respectively, according to the input gas used with the N₂ + O₂ mixture being the most effective. Most APP jets reduced the numbers of aerobic bacteria on the meat surfaces to <10² CFU/g, and the numbers remained below that level of detection after storage at 10 °C for 7 days. The results indicate that APP jets are effective for the inactivation of L. monocytogenes on sliced meats and for prolonging the shelf-life of such foods.     

Inactivation of bacteriophages by thermal and high-pressure treatment

Dairy companies commonly experience fermentation failures due to bacteriophages that are spread mainly by milk, whey or air. Heat or high-pressure treatment may potentially reduce the phage titre, but further knowledge about the inactivation kinetics is desirable. Inactivation experiments were carried out with the commonly occurring lactococcal phages P001 and P008. Phage suspensions in calcium-enriched M17-broth were heated at 55–80 °C, or high-pressure treated at up to 600 MPa. Kinetic analysis showed that the order of inactivation reaction was above 1; thus, inactivation kinetics were approximated by a non-linear regression model. The Arrhenius parameters, rate constant, k_p,T, and activation energy, E_A (for heat treatments), and the volume of activation, ΔV^# (for pressure treatments) were calculated. Both measured and calculated results indicate that phage P008 was the more heat- and pressure-resistant of the two. By combining the results from heat and pressure inactivations, a pressure–temperature diagram for phage P008 was established.     

Antiviral activities of Artemisia princeps var. orientalis essential oil and its α-thujone against norovirus surrogates

Food Science and Biotechnology - Artemisia princeps var. orientalis is a well-known medicinal food, which has been used for the treatment of several diseases including bacterial infection. We...     

Effect of cultural system and essential oil treatment on antioxidant capacity in raspberries

The effects of cultural system and essential oil treatment on antioxidant capacities in raspberries were evaluated. Raspberries were hand-harvested from organic and conventional farms in Maryland, USA, and were treated with essential oil including carvacrol, anethole, cinnamic acid, perillaldehyde, cinnamaldehyde, and linalool. Results from this study showed that raspberries grown from organic culture exhibited higher value of antioxidant capacities and individual flavonoids contents. Moreover, the organic culture also enhanced the activities of antioxidant enzymes. In addition, essential oil treatments promoted the antioxidant enzymes activities and antioxidant capacities of raspberries, and the most effective compound was perillaldehyde. In conclusion, raspberries produced from organic culture contained significantly higher antioxidant capacities than those produce from conventional culture. Postharvest essential oil treatments have positive effect on enhancing antioxidant capacities in raspberries from both organic and conventional cultures.     

Inactivation of Salmonella on pecan nutmeats by hot air treatment and oil roasting

Studies were done to determine the effectiveness of hot air drying, dry roasting, and oil roasting in killing Salmonella on pecan nutmeats. Pecan halves and pieces were inoculated by immersion in a five-serotype suspension of Salmonella or by surface application of powdered chalk containing the pathogen. Hot air treatment of low-moisture (2.8 to 4.1%) and high-moisture (10.5 to 11.2%) immersion-inoculated nutmeats (initial population, 6.18 to 7.16 log CFU/g) at 120°C for 20 min reduced the number of Salmonella by 1.18 to 1.26 and 1.89 to 2.04 log CFU/g, respectively. However, regardless of the moisture content, hot air treatment of pecan halves containing 0.77 log CFU/g at 120°C for 20 min failed to eliminate Salmonella. Reductions were >7 log CFU/g when dry pieces were dry roasted at 160°C for 15 min. Treatment of halves at 140°C for 20 min, 150°C for 15 min, or 170°C for 10 min reduced Salmonella by 5 log CFU/g. The pathogen was slightly more heat resistant in immersion-inoculated nutmeats than on surface-inoculated nutmeats. Exposure of immersion-inoculated pieces to peanut oil at 127°C for 1.5 min or 132°C for 1.0 min reduced the number of Salmonella by 5 log CFU/g. Treatment of halves at 138°C for 2.0 min reduced Salmonella by 5 log CFU/g; treatment at 132°C for 2.5 to 4.0 min did not always achieve this reduction. Hot air treatment cannot be relied upon to reduce Salmonella by 5 log CFU/g of raw pecan nutmeats without changing sensory qualities. Treatment temperatures and times typically used to oil roast nutmeats appear to be sufficient to reduce Salmonella by 5 log CFU/g.     

Inactivation of staphylococcal enterotoxins by heat and reactivation by high pH treatment

Inactivation of staphylococcal enterotoxins (SE) added to different media upon heat treatment (80 degrees C or 100 degrees C for 10 min) and reactivation of inactivated SE was studied. In gelatin (pH 4.0), lettuce extract, peas and beans extracts and ovalbumin (pH 5.0) the immunological activity of SE was lost almost completely upon heating. The loss of immunological activity of SEA was accompanied by a concomitant loss of biological activity of this toxin (monkey feeding test). A high pH treatment (pH 11) of the different preparations restored both the immunological and biological activity in most samples tested. Heating at 80 degrees C or 100 degrees C for 10 min of SE containing gelatin (pH 7.0), cauliflower extract (pH 4.0 or pH 7.0), milk (pH 4.0), casein (pH 6.0), rice extract (pH 7.0), noodles extract (pH 4.0) and oat-flakes extract (pH 7.0) had a much lower effect on the immunological activity of the SE (activity greater than or equal to 25%).     

Survival of calicivirus in foods and on surfaces: experiments with feline calicivirus as a surrogate for norovirus

Although there is a large body of evidence incriminating foods as vehicles in the transmission of norovirus, little is known about virus survival in foods and on surfaces. Feline calicivirus was used as a surrogate for norovirus to investigate its survival in representative foods of plant and animal origin and on metal surfaces. Known concentrations of feline calicivirus in a natural fecal suspension were deposited onto lettuce, strawberries, ham, or stainless steel and incubated for 7 days at refrigeration or room temperatures. Virus was recovered at 1-day intervals, and the titers of the virus were determined by plaque assay. Infectious virus was recoverable until day 7 from lettuce, ham, and stainless steel. Statistically higher titers of feline calicivirus (P < 0.05) were recovered from ham under all conditions than from lettuce, strawberries, or stainless steel. These data provide valuable information for epidemiological and monitoring purposes as well as for the development of food processing practices and appropriate strategies to inactivate norovirus and control its transmission via foods and surfaces.