RESUMEN
In this study the suitability of a thin-film reactor (TFR) equipped with special flow guiding elements (FGE) was examined to analyse its capability to inactivate microorganisms in milk. Experiments were carried out with UHT-milk inoculated with Escherichia coli (E. coli), DH5α and Listeria innocua (L. innocua) WS 2258. Furthermore, the inactivation of microorganisms originally occurring in raw milk was investigated. E. coli, DH5α and L. innocua serving as biodosimeter were reduced by 4.58-log and 3.19-log, respectively. In milk, the original microorganisms showed a 4-log reduction. Without FGE the reduction was below 0.13-log. Thus, it can be derived that the efficacy of a UV-C thin-film reactor processing absorptive media like milk can be highly improved using FGE.
Asunto(s)
Escherichia coli/efectos de la radiación , Irradiación de Alimentos/métodos , Listeria/efectos de la radiación , Leche/microbiología , Animales , Recuento de Colonia Microbiana , Escherichia coli/crecimiento & desarrollo , Irradiación de Alimentos/instrumentación , Microbiología de Alimentos , Listeria/crecimiento & desarrollo , Microbiota/efectos de la radiación , Leche/química , Rayos UltravioletaRESUMEN
We aimed to study the efficacy of a water-assisted UVC light device (WUVC) as an innovative clean technology for the disinfection of fresh sound tomatoes and processing wash water and water turbidity was evaluated as a critical parameter. First, wash waters with different turbidities (from 0.4 to 828 NTU) were inoculated with Listeria innocua and treated in the WUVC device at different dosages. Secondly, fresh tomatoes, inoculated with L. innocua and non-inoculated ones, were treated using the WUVC device containing wash water of different turbidities for different times. The reduction of L. innocua populations on wash water and on the surface of tomato was influenced by turbidity; lower reduction values were observed at higher turbidities. Washing tomatoes with tap water with UVC lamps off (control treatment, TW) decreased L. innocua population on the surface of tomatoes but did not eliminate those bacteria that went into the water. Contrarily, when UVC lights were on, L. innocua population in wash water after treatment significantly decreased, those in clean water being the lowest populations. Reductions of native microbiota on the clean water treated with the highest UV-C radiation dose were lower than those obtained when tomatoes were artificially inoculated. We demonstrated that high reductions of L. innocua population on fresh tomatoes could be achieved using the WUVC system but some drawbacks related to the increase of turbidity should be solved for its implementation in real conditions.
Asunto(s)
Desinfección/métodos , Irradiación de Alimentos/métodos , Listeria/efectos de la radiación , Solanum lycopersicum/microbiología , Recuento de Colonia Microbiana , Desinfección/instrumentación , Frutas/microbiología , Listeria/crecimiento & desarrollo , Rayos Ultravioleta , Agua/químicaRESUMEN
HIGHLIGHTS: Thermal and nonthermal methods can support a 5-log CFU reduction of model bacteria introduced into tiger nut milk. Thermal treatment of tiger nut milk results in significant loss of protein, antioxidants, and quality properties. HHP or UV-C treatment of tiger nut milk retains quality and nutritional characteristics. HHP or UV-C are suitable for the pasteurization of tiger nut milk.
Asunto(s)
Fenómenos Fisiológicos Bacterianos , Microbiología de Alimentos , Calor , Presión Hidrostática , Viabilidad Microbiana , Valor Nutritivo , Productos Vegetales , Animales , Fenómenos Fisiológicos Bacterianos/efectos de la radiación , Escherichia coli/fisiología , Escherichia coli/efectos de la radiación , Listeria/fisiología , Listeria/efectos de la radiación , Salmonella/fisiología , Salmonella/efectos de la radiación , Rayos Ultravioleta , Productos Vegetales/microbiologíaRESUMEN
There is a critical unmet need to improve microbial safety of fresh fruits and vegetables. Current sanitation approaches cannot achieve >2 log inactivation of bacteria on fresh produce. Thus, there is a need to develop antimicrobial strategies that can consistently achieve >2 logs of bacterial inactivation on the surface of diverse fresh produce. Furthermore it is highly desired that these antimicrobial strategies have reduced environmental impact and are clean label solutions for food products. In this study, we evaluated the combination of curcumin and UV-A light radiation for the inactivation of inoculated E. coli O157:H7 and L. innocua bacterial cells on the surface of spinach, lettuce and tomatoes. Curcumin was deposited on the surface of fresh produce by either aerosolization or conventional spray-atomization methods before exposing the contaminated produce to UV-A light for 10â¯min (total light fluence of 20.4â¯kJâ¯m-2). Results showed that the proposed combination of aerosolized or sprayed curcumin and UV-A light radiation can reduce the initial Escherichia coli O157:H7 and Listeria innocua load from 6 log CFU cm-2 to approximately 3 log CFU cm-2 on spinach, lettuce and tomato surfaces. Furthermore, there was no significant difference in bacterial reduction between the different types of inoculated fresh produce surfaces (Pâ¯>â¯.05). Interestingly, subsequent curcumin deposition and UV-A light exposure cycles were not able to further reduce the bacterial load below the observed threshold of approximately 3 log CFU cm-2. Lastly, the combination of aerosolized curcumin and UV-A light radiation did not affect the color or the texture of the treated fresh produce samples. The findings described in this study illustrate the potential of applying aerosolized or sprayed curcumin under UV-A light illumination to improve microbial safety of fresh produce products.
Asunto(s)
Antibacterianos/farmacología , Curcumina/farmacología , Escherichia coli O157 , Lactuca/microbiología , Listeria , Rayos Ultravioleta , Aerosoles , Escherichia coli O157/efectos de los fármacos , Escherichia coli O157/efectos de la radiación , Manipulación de Alimentos , Conservación de Alimentos , Listeria/efectos de los fármacos , Listeria/efectos de la radiaciónRESUMEN
This study investigated the efficacy of power ultrasound (US) for the inactivation of Escherichia coli and Listeria innocua in the presence of sodium salt and salt replacers. Inoculated bacteria suspensions were treated at ultrasonic frequencies of 33 or 20â¯kHz alone or in combination, and in the presence of 5% NaCl, 5% KCl or 5% NaCl/KCl. Inactivation curves were fitted to the Weibull and the Biphasic models. The goodness of the fit for each model was evaluated based on R2 and RMSE, while AIC and BIC values were used to choose the best model predictor. The Weibull and the biphasic models showed high regression coefficient (R2â¯>â¯0.99) and low RMSE (<0.03) values. According to the results, inactivation up to 6 log for E. coli K12 and to 4 log for L. innocua could be achieved within one hour of ultrasound treatment. However, the presence of NaCl, or its substitution with KCl did not affect the degree of inhibition for both microorganisms. The results of this study suggest that power ultrasound treatment may be employed for the inactivation of microorganisms when low salt or salt substitutes are employed.
Asunto(s)
Escherichia coli K12/efectos de la radiación , Ondas de Choque de Alta Energía , Listeria/efectos de la radiación , Recuento de Colonia Microbiana , Escherichia coli K12/efectos de los fármacos , Escherichia coli K12/aislamiento & purificación , Listeria/efectos de los fármacos , Listeria/aislamiento & purificación , Modelos Biológicos , Cloruro de Potasio/farmacología , Cloruro de Sodio/farmacologíaRESUMEN
The effect of pulsed light treatment on the lag phase and the maximum specific growth rate of Listeria innocua was determined in culture media at 7⯰C. Fluences of 0.175, 0.350 and 0.525â¯J/cm2 were tested. The lag phase of the survivors increased as fluence did, showing significant differences for all the doses; an 8.7-fold increase was observed at 0.525â¯J/cm2. Pulsed light decreased the maximum specific growth rate by 38% at the same fluence. Both parameters were also determined by time-lapse microscopy at 25⯰C in survivors to 0.525â¯J/cm2, with an increase of 13-fold of the lag phase and a 45% decrease of the maximum specific growth rate. The higher the fluence, the higher the variability of both parameters was. To characterize pulsed light damage on L. innocua, the formation of dimers on DNA was assessed, and a proteomic study was undertaken. In cells treated with 0.525â¯J/cm2, cyclobutane pyrimidine dimers and pyrimidine (6-4) pyrimidone photoproducts were detected at 5:1 ratio. Pulsed light induced the expression of three proteins, among them the general stress protein Ctc. Furthermore, treated cells showed an up-regulation of proteins related to metabolism of nucleotides and fatty acids, as well as with translation processes, whereas flagellin and some glucose metabolism proteins were down-regulated. Differences in the proteome of the survivors could contribute to explain the mechanisms of adaptation of L. innocua after pulsed light treatment.
Asunto(s)
ADN Bacteriano/efectos de la radiación , Luz , Listeria , Proteoma/efectos de la radiación , Flagelina/biosíntesis , Listeria/crecimiento & desarrollo , Listeria/metabolismo , Listeria/efectos de la radiación , ProteómicaRESUMEN
The antimicrobial efficacy of 400 nm photoirradiated caffeic acid (CA, 5 mM) was evaluated against Escherichia coli O157:H7 and Listeria innocua. A stronger antimicrobial effect was observed on E. coli than on L. innocua where the combined treatment resulted in 4 and 1 log(CFU/mL) reductions, respectively. The treatment's effects on cellular metabolism (resazurin assay), uptake of CA (fluorescence technique) and membrane damage (propidium iodide assay) were studied in both species. CA uptake increased in both species, but membrane damage was only observed in E. coli O157:H7. The treatment had minimal impact on metabolic activity in both species. The treatment applied to the surface of spinach leaves was found to be effective against E. coli O157:H7. The novel treatment proposed in this study has the potential to improve the microbial food safety of fresh produce.
Asunto(s)
Antibacterianos/farmacología , Ácidos Cafeicos/farmacología , Escherichia coli O157/efectos de los fármacos , Escherichia coli O157/efectos de la radiación , Conservación de Alimentos/métodos , Listeria/efectos de los fármacos , Listeria/efectos de la radiación , Spinacia oleracea/microbiología , Recuento de Colonia Microbiana , Escherichia coli O157/crecimiento & desarrollo , Contaminación de Alimentos/prevención & control , Irradiación de Alimentos/métodos , Conservación de Alimentos/instrumentación , Listeria/crecimiento & desarrollo , Hojas de la Planta/microbiologíaRESUMEN
The synergistic combination of the food colorant Erythrosin B (E-B, FD&C 3) (0, 25, and 50µM) and low-frequency ultrasound (20kHz, 0.86-0.90WmL-1) was evaluated against Listeria innocua. Although E-B was antibacterial by itself, the inactivation rate significantly increased in a concentration-dependent manner upon exposure to ultrasound and followed a sigmoidal behavior. The enhanced antimicrobial effect of E-B in the presence of ultrasound can be explained in part from a microbubble disappearance study in which it was confirmed that the presence of E-B enhances inertial cavitation, thereby enhancing the antimicrobial effect of ultrasound. The inactivation rate in a sequential treatment, where L. innocua was sonicated for 4min followed by exposure to 25µM Erythrosin B, was comparable to that obtained by the simultaneous treatment, indicating complementary mechanisms of inactivation. Fluorescence microscopy showed attachment of E-B to the cells, which may explain its intrinsic antimicrobial property. Other mechanism may include the confirmed decrease in the cavitation threshold of water by addition of E-B, resulting in more effective cavitation. The study offers a proof-of-concept of a novel approach to complement ultrasound treatment for enhanced microbial inactivation.
Asunto(s)
Antiinfecciosos , Eritrosina , Ondas Ultrasónicas , Antiinfecciosos/farmacología , Antiinfecciosos/efectos de la radiación , Eritrosina/farmacología , Eritrosina/efectos de la radiación , Listeria/efectos de los fármacos , Listeria/efectos de la radiación , Microscopía FluorescenteRESUMEN
AIMS: Pulsed light (PL) is an upcoming nonthermal decontamination technology mainly used for surface sterilization. The objective of this study was to investigate the extent of cellular damage caused by PL treatments of Listeria innocua and Escherichia coli on a polysaccharide surface in order to gain knowledge about the main inactivation pathways. METHODS AND RESULTS: The impact of PL on the cellular ATP level was investigated as well as the bacterial ability to take up fluorescently labelled glucose (2-NBDG). Furthermore, the extent of DNA damages was assessed by qPCR. The ability of L. innocua and E. coli to photorepair under artificial daylight exposure was quantified. Finally, the induction of reactive oxygen species (ROS) and lipid peroxidation were studied by fluorometric detection of ROS and thiobarbituric acid reactive substances (TBARS). It is shown that intracellular ATP levels and glucose uptake ability do not correlate with the immediate loss of bacterial reproducibility, which indicates that cellular activity and energy may remain on a relatively high level, although growth on tryptic soy agar is not observable. Sequence specific investigation of PL induced DNA damages by qPCR revealed distinct differences between L. innocua and E. coli although the observed inactivation efficacy of PL by the culture based method was similar. Photoreactivation has been observed for both bacteria, a higher recovery rate of up to 2 log was seen in case of E. coli. Intracellular ROS and lipid peroxides were both detectable at relatively high fluencies with E. coli so the contribution of oxidative damage to microbial inactivation of PL cannot be excluded. CONCLUSIONS: Escherichia coli as well as L. innocua cells have proven to maintain residual cellular activity after having been exposed to PL even when they are not able to reproduce any more. High proportions of sublethal damages were also obvious with regard to occurring photoreactivation. The destruction of bacterial DNA seems to be the primary mechanism of inactivation of PL but the involvement of other factors like oxidative stress cannot be excluded. SIGNIFICANCE AND IMPACT OF THE STUDY: The observed data underline that bacteria are not immediately inactivated after exposure to PL as different indicators of cellular energy are still detectable even when cells do not reproduce on solid media any more. DNA is the primary target of PL, but as the extent of damage among different bacteria may not reveal their actual sensitivity, other destructive effects should also be considered.
Asunto(s)
Descontaminación/métodos , Escherichia coli/efectos de la radiación , Listeria/efectos de la radiación , Escherichia coli/crecimiento & desarrollo , Luz , Listeria/crecimiento & desarrollo , Viabilidad Microbiana/efectos de la radiaciónRESUMEN
Cheese products are susceptible to postprocessing cross-contamination by bacterial surface contamination during slicing, handling, or packaging, which can lead to food safety issues and significant losses due to spoilage. This study examined the effectiveness of pulsed-light (PL) treatment on the inactivation of the spoilage microorganism Pseudomonas fluorescens, the nonenterohemorrhagic Escherichia coli ATCC 25922 (nonpathogenic surrogate of Escherichia coli O157:H7), and Listeria innocua (nonpathogenic surrogate of Listeria monocytogenes) on cheese surface. The effects of inoculum level and cheese surface topography and the presence of clear polyethylene packaging were evaluated in a full factorial experimental design. The challenge microorganisms were grown to early stationary phase and subsequently diluted to reach initial inoculum levels of either 5 or 7 log cfu/slice. White Cheddar and process cheeses were cut into 2.5×5 cm slices, which were spot-inoculated with 100 µL of bacterial suspension. Inoculated cheese samples were exposed to PL doses of 1.02 to 12.29 J/cm(2). Recovered survivors were enumerated by standard plate counting or the most probable number technique, as appropriate. The PL treatments were performed in triplicate and data were analyzed using a general linear model. Listeria innocua was the least sensitive to PL treatment, with a maximum inactivation level of 3.37±0.2 log, followed by P. fluorescens, with a maximum inactivation of 3.74±0.8 log. Escherichia coli was the most sensitive to PL, with a maximum reduction of 5.41±0.1 log. All PL inactivation curves were nonlinear, and inactivation reached a plateau after 3 pulses (3.07 J/cm(2)). The PL treatments through UV-transparent packaging and without packaging consistently resulted in similar inactivation levels. This study demonstrates that PL has strong potential for decontamination of the cheese surface.
Asunto(s)
Queso/microbiología , Luz , Animales , Recuento de Colonia Microbiana , Descontaminación/métodos , Relación Dosis-Respuesta a Droga , Escherichia coli O157/aislamiento & purificación , Escherichia coli O157/efectos de la radiación , Contaminación de Alimentos , Microbiología de Alimentos , Embalaje de Alimentos , Listeria/aislamiento & purificación , Listeria/efectos de la radiación , Listeria monocytogenes/aislamiento & purificación , Listeria monocytogenes/efectos de la radiación , Polietileno/químicaRESUMEN
Inactivation of Listeria innocua by pulsed light (PL) was evaluated at different post-treatment temperature and illumination conditions. The impact of post-PL-treatment temperature on L. innocua culturability was evaluated for cells cultured at 37 °C (optimal growth temperature) and 4 °C (classical refrigerated food temperature). For both culture conditions, significant higher reductions (up to 3 log) were observed after post-PL-treatment temperature of 4 °C than of 37 °C. Contrarily, L. innocua culturability after PL treatment increased up to 2.2 log in presence of daylight illumination in comparison to dark storage. This photorepair mechanism was quickly activated reaching the maximum photoreactivation level after only 30 min of illumination. Moreover, photorepair capacity was rapidly reduced by increasing the time in darkness from PL treatment to samples illumination, being completely lost after time in darkness equal or greater than 5 h. According to these findings, the combination of PL with post-treatment temperature of 4 °C has a synergistic effect on the inactivation of L. innocua, whereas post-treatment daylight illumination has an antagonic effect on PL antimicrobial efficacy. Post-PL-treatment temperature and illumination conditions could be thereby considered important environmental factors to activate, inhibit or control the repair and/or growth of L. innocua survivors after PL treatment.
Asunto(s)
Irradiación de Alimentos/métodos , Listeria/efectos de la radiación , Viabilidad Microbiana/efectos de la radiación , Irradiación de Alimentos/instrumentación , Luz , Listeria/crecimiento & desarrollo , Temperatura , Factores de TiempoRESUMEN
AIMS: The application of broad-spectrum intense light pulses is an innovative nonthermal technology for the decontamination of packaging materials, liquids or foodstuffs. The objective of this study was the fundamental investigation of the cellular impact of a pulsed light treatment on Listeria innocua and Escherichia coli. METHODS AND RESULTS: Flow cytometry in combination with different fluorescent stains, conventional plate count technique and a viability assay were applied to investigate the effects of a pulsed light treatment on the physiological properties of L. innocua and E. coli. The results showed that loss of cultivability occurred at considerably lower fluences than the shutdown of cellular functions such as the depolarization of cell membranes, the loss of metabolic, esterase and pump activities or the occurrence of membrane damage. Therefore, a considerable proportion of cells appeared to have entered the viable but nonculturable (VBNC) state after the pulsed light treatment. A high percentage of L. innocua was able to maintain certain cellular vitality functions after storage overnight, whereas a further decrease in vitality was observed in case of E. coli. The loss of culturability was on the other hand directly accompanied by the formation of reactive oxygen species (ROS) and DNA damages, which were assessed by the ROS-sensitive probe DCFH-DA and RAPD-PCR, respectively. CONCLUSIONS: A significant discrepancy between conventional plate counts and different viability staining parameters was observed, which shows that a pulsed light treatment does not cause an immediate shutdown of vitality functions even when the number of colony-forming units already decreased for more than 6 log10 sample(-1) . Oxidative stress with concomitant damage to the DNA molecule showed to be directly responsible for the loss of cultivability due to pulsed light rather than a direct rupture of cell membranes or inactivation of intracellular enzymes. SIGNIFICANCE AND IMPACT OF THE STUDY: The presented results suggest an UV light-induced photochemical rather than a photothermal or photophysical inactivation of bacterial cells by pulsed light under the applied experimental conditions. Flow cytometry in combination with different viability stains proved to be a suitable technique to gain deeper insight into the cellular response of bacteria to inactivation processes like a pulsed light treatment.
Asunto(s)
Descontaminación , Escherichia coli/efectos de la radiación , Luz , Listeria/efectos de la radiación , Daño del ADN , Escherichia coli/crecimiento & desarrollo , Escherichia coli/metabolismo , Escherichia coli/fisiología , Citometría de Flujo , Listeria/crecimiento & desarrollo , Listeria/metabolismo , Listeria/fisiologíaRESUMEN
The objective of this study was to produce phage display-derived binders with the ability to distinguish Listeria monocytogenes from other Listeria spp., which may have potential utility to enhance detection of Listeria monocytogenes. To obtain binders with the desired binding specificity a series of surface and solution phage-display biopannings were performed. Initially, three rounds of surface biopanning against gamma-irradiated L. monocytogenes serovar 4b cells were performed followed by an additional surface biopanning round against L. monocytogenes 4b which included prior subtraction biopanning against gamma-irradiated L. innocua cells. In an attempt to further enhance binder specificity for L. monocytogenes 4b two rounds of solution biopanning were performed, both rounds included initial subtraction solution biopanning against L. innocua. Subsequent evaluations were performed on the phage clones by phage binding ELISA. All phage clones tested from the second round of solution biopanning had higher specificity for L. monocytogenes 4b than for L. innocua and three other foodborne pathogens (Salmonella spp., Escherichia coli and Campylobacter jejuni). Further evaluation with five other Listeria spp. revealed that one phage clone in particular, expressing peptide GRIADLPPLKPN, was highly specific for L. monocytogenes with at least 43-fold more binding capability to L. monocytogenes 4b than to any other Listeria sp. This proof-of-principle study demonstrates how a combination of surface, solution and subtractive biopanning was used to maximise binder specificity. L. monocytogenes-specific binders were obtained which could have potential application in novel detection tests for L. monocytogenes, benefiting both the food and medical industries.
Asunto(s)
Antígenos Bacterianos/química , Técnicas de Tipificación Bacteriana/métodos , Ensayo de Inmunoadsorción Enzimática/métodos , Listeria monocytogenes/aislamiento & purificación , Biblioteca de Péptidos , Secuencia de Aminoácidos , Antígenos Bacterianos/metabolismo , Campylobacter jejuni/química , Escherichia coli/química , Rayos gamma , Listeria/química , Listeria/efectos de la radiación , Listeria monocytogenes/química , Listeria monocytogenes/efectos de la radiación , Datos de Secuencia Molecular , Unión Proteica , Salmonella/química , Especificidad de la EspecieRESUMEN
Pulsed light (PL) treatment can effectively inactivate a large proportion of contaminating bacteria on surfaces and in clear solutions. An important issue that needs to be investigated is whether repeated exposure to PL treatment causes any changes to the growth and resistance behavior of the bacteria surviving the treatment. To test this, three challenge microorganisms were used: Listeria monocytogenes, Listeria innocua, and Escherichia coli. Cells of the challenge bacteria were treated with either low or high PL doses. Survivors of the PL treatment were enumerated, isolated, regrown, and exposed again to PL treatment. PL inactivation curves were generated for the survivors of each exposure cycle (as well as controls) to examine possible differences induced by repeated treatments. Growth curves of L. monocytogenes, L. innocua, and E. coli isolates recovered from exposure to either 1.1 or 10.1 J/cm(2) were not significantly different from the growth curves of untreated cells. Reduction levels of up to 4 and up to 6 log CFU were obtained after exposure to 1.1 and 10.1 J/cm(2), respectively, both for the controls and the repeatedly treated and recovered isolates. These results show that PL did not significantly change the growth kinetics or resistance to PL of the target microorganisms after up to 10 exposures. These findings have significance for the practical application of PL treatment, as they indicate that this technology does not select for microorganisms with increased resistance.
Asunto(s)
Escherichia coli/crecimiento & desarrollo , Irradiación de Alimentos , Conservación de Alimentos/métodos , Listeria monocytogenes/crecimiento & desarrollo , Listeria/crecimiento & desarrollo , Recuento de Colonia Microbiana , Seguridad de Productos para el Consumidor , Relación Dosis-Respuesta en la Radiación , Escherichia coli/efectos de los fármacos , Microbiología de Alimentos , Humanos , Cinética , Luz , Listeria/efectos de la radiación , Listeria monocytogenes/efectos de la radiación , Modelos Biológicos , Rayos UltravioletaRESUMEN
The effect of ß-lactoglobulin and ß-casein on the pulsed light (PL) inactivation of Listeria innocua was evaluated. For low protein concentrations (ß-lactoglobulin and ß-casein up to 10 mg/mL), the lowest fluences applied (0.2 J/cm(2)) induced more than 7 Log reductions in cell counts. However, higher fluences were needed to induce similar reduction in L. innocua counts when this bacterium was suspended in solutions of higher protein concentration. The fluence required to induce similar microbial inactivation was lower in ß-casein than in ß-lactoglobulin solutions. For all protein solutions, the inactivation curves followed first order kinetics. The specific inactivation rate for L. innocua inactivation in protein solutions depended on the quantity of light transmitted in the range 230-290 nm by protein solutions. This work shows that PL technology could be used for the decontamination of high protein content solutions like whey or even higher protein concentration solutions.
Asunto(s)
Caseínas/farmacología , Lactoglobulinas/farmacología , Listeria/efectos de los fármacos , Listeria/efectos de la radiación , Rayos Ultravioleta , Descontaminación/métodos , Microbiología de Alimentos , Conservación de Alimentos , Proteínas de la Leche/farmacología , Soluciones , Proteína de Suero de LecheRESUMEN
UNLABELLED: Recent outbreaks associated to the consumption of raw or minimally processed vegetable products that have resulted in several illnesses and a few deaths call for urgent actions aimed at improving the safety of those products. Electron beam irradiation can extend shelf-life and assure safety of fresh produce. However, undesirable effects on the organoleptic quality at doses required to achieve pathogen inactivation limit irradiation. Ways to increase pathogen radiation sensitivity could reduce the dose required for a certain level of microbial kill. The objective of this study was to evaluate the effectiveness of using natural antimicrobials when irradiating fresh produce. The minimum inhibitory concentration of 5 natural compounds and extracts (trans-cinnamaldehyde, eugenol, garlic extract, propolis extract, and lysozyme with ethylenediaminetetraacetate acid (disodium salt dihydrate) was determined against Salmonella spp. and Listeria spp. In order to mask odor and off-flavor inherent of several compounds, and to increase their solubility, complexes of these compounds and extracts with ß-cyclodextrin were prepared by the freeze-drying method. All compounds showed bacteriostatic effect at different levels for both bacteria. The effectiveness of the microencapsulated compounds was tested by spraying them on the surface of baby spinach inoculated with Salmonella spp. The dose (D10 value) required to reduce the bacterial population by 1 log was 0.190 kGy without antimicrobial addition. The increase in radiation sensitivity (up to 40%) varied with the antimicrobial compound. These results confirm that the combination of spraying microencapsulated antimicrobials with electron beam irradiation was effective in increasing the killing effect of irradiation. PRACTICAL APPLICATION: Foodborne illness outbreaks attributed to fresh produce consumption have increased and present new challenges to food safety. Current technologies (water washing or treating with 200 ppm chlorine) cannot eliminate internalized pathogens. Ionizing radiation is a viable alternative for eliminating pathogens; however, the dose required to inactivate these pathogens is often too high to be tolerated by the fresh produce without undesirable quality changes. This study uses natural antimicrobial ingredients as radiosensitizers. These ingredients were encapsulated and applied to fresh produce that was subsequently irradiated. The process results in high level of microorganism inactivation using lower doses than the conventional irradiation treatments.
Asunto(s)
Antibacterianos/farmacología , Irradiación de Alimentos , Listeria/efectos de la radiación , Hojas de la Planta/microbiología , Fármacos Sensibilizantes a Radiaciones/farmacología , Salmonella/efectos de la radiación , Spinacia oleracea/microbiología , Antibacterianos/química , Antibacterianos/metabolismo , Antiinfecciosos/química , Antiinfecciosos/metabolismo , Antiinfecciosos/farmacología , Productos Biológicos/química , Productos Biológicos/metabolismo , Productos Biológicos/farmacología , Rastreo Diferencial de Calorimetría , Portadores de Fármacos/química , Portadores de Fármacos/farmacología , Composición de Medicamentos , Electrones , Irradiación de Alimentos/efectos adversos , Embalaje de Alimentos , Enfermedades Transmitidas por los Alimentos/prevención & control , Liofilización , Listeria/efectos de los fármacos , Listeria/crecimiento & desarrollo , Listeria/aislamiento & purificación , Pruebas de Sensibilidad Microbiana , Viabilidad Microbiana/efectos de los fármacos , Viabilidad Microbiana/efectos de la radiación , Hojas de la Planta/crecimiento & desarrollo , Hojas de la Planta/efectos de la radiación , Fármacos Sensibilizantes a Radiaciones/química , Fármacos Sensibilizantes a Radiaciones/metabolismo , Salmonella/efectos de los fármacos , Salmonella/crecimiento & desarrollo , Salmonella/aislamiento & purificación , Solubilidad , Spinacia oleracea/crecimiento & desarrollo , Spinacia oleracea/efectos de la radiación , beta-Ciclodextrinas/químicaRESUMEN
The FDA recently approved irradiation treatment of leafy greens such as spinach up to 1 kGy; however, it is important to reduce the dose required to decontaminate the produce while maintaining its quality. Thus, the objectives of this study were: (1) to assess the radiation sensitivities of Salmonella spp. and Listeria spp. inoculated in ready-to-eat baby spinach leaves under modified atmosphere packaging (MAP) and irradiated using a 1.35-MeV Van de Graff accelerator (the leaves were irradiated both at room temperature and at -5 °C); and (2) to understand and optimize the synergistic effect of MAP and irradiation by studying the radiolysis of ozone formation under different temperatures, the effect of dose rate on its formation, and its decomposition. Results showed that increased concentrations of oxygen in the packaging significantly increased the radiation sensitivity of the test organisms, ranging from 7% up to 25% reduction in D(10)-values. In particular, radiosensitization could be effected (P < 0.05) by production of ozone, which increases with increasing dose-rate and oxygen concentration, and reducing temperatures. Radiosensitization was demonstrated for both microorganisms with irradiation of either fresh or frozen (-5 °C) baby spinach. These results suggest that low-dose (below 1 kGy) e-beam radiation under modified atmosphere packaging (100% O(2) and N(2):O(2)[1:1]) may be a viable tool for reducing microbial populations or eliminating Salmonella spp. and Listeria spp. from baby spinach. A suggested treatment to achieve a 5-log reduction of the test organisms would be irradiation at room temperature under 100% O(2) atmosphere at a dose level of 0.7 kGy. Practical Application: Decontamination of minimally processed fruits and vegetables from food-borne pathogens presents technical and economical challenges to the produce industry. Internalized microorganisms cannot be eliminated by the current procedure (water-washed or treated with 200-ppm chlorine). The only technology available commercially is ionizing radiation; however, the actual radiation dose required to inactivate pathogens is too high to be tolerated by the product without unwanted changes. This study shows a new approach in using MAP with 100% O(2), which is converted to ozone to radiosensitize pathogens while improving the shelf life of minimally processed fruits and vegetables. The process results in a high level of microorganism inactivation using lower doses than the conventional irradiation treatments.
Asunto(s)
Comida Rápida/microbiología , Irradiación de Alimentos , Listeria/efectos de la radiación , Hojas de la Planta/microbiología , Salmonella/efectos de la radiación , Spinacia oleracea/microbiología , Frío , Recuento de Colonia Microbiana , Relación Dosis-Respuesta en la Radiación , Electrones , Comida Rápida/análisis , Comida Rápida/efectos de la radiación , Irradiación de Alimentos/métodos , Embalaje de Alimentos/métodos , Alimentos Congelados/análisis , Alimentos Congelados/microbiología , Alimentos Congelados/efectos de la radiación , Hidrólisis , Viabilidad Microbiana/efectos de la radiación , Oxígeno/farmacología , Oxígeno/efectos de la radiación , Ozono/análisis , Ozono/farmacología , Ozono/efectos de la radiación , Hojas de la Planta/química , Hojas de la Planta/efectos de la radiación , Control de Calidad , Fármacos Sensibilizantes a Radiaciones/análisis , Fármacos Sensibilizantes a Radiaciones/farmacología , Fármacos Sensibilizantes a Radiaciones/efectos de la radiación , Spinacia oleracea/química , Spinacia oleracea/efectos de la radiaciónRESUMEN
High Intensity Light Pulses (HILP) represent an emerging processing technology which uses short (100-400 µs) light pulses (200-1100 nm) for product decontamination. In this study, model and real foods of differing transparencies (maximum recovery diluent (MRD), apple and orange juices and milk) were exposed to HILP in a batch system for 0, 2, 4 or 8 s at a frequency of 3 Hz. After treatment, inactivation of Escherichia coli or Listeria innocua was evaluated in pre-inoculated samples. Sensory and other quality attributes (colour, pH, Brix, titratable acidity, non-enzymatic browning, total phenols and antioxidant capacity (TEAC)) were assessed in apple juice. Microbial kill decreased with decreasing transparency of the medium. In apple juice (the most transparent beverage) E. coli decreased by 2.65 and 4.5 after exposure times of 2 or 4 s, respectively. No cell recovery was observed after 48 h storage at 4°C. No significant differences were observed in quality parameters, excepting TEAC and flavour score, where 8 s exposure caused a significant decrease (p<0.05). Based on these results, HILP with short exposure times could represent a potential alternative to thermal processing to eliminate undesirable microorganisms, while maintaining product quality, in transparent fruit juices.
Asunto(s)
Bebidas/microbiología , Escherichia coli/crecimiento & desarrollo , Contaminación de Alimentos/prevención & control , Luz , Listeria/crecimiento & desarrollo , Animales , Citrus sinensis/microbiología , Recuento de Colonia Microbiana , Seguridad de Productos para el Consumidor , Medios de Cultivo Condicionados , Escherichia coli/efectos de la radiación , Manipulación de Alimentos/métodos , Microbiología de Alimentos , Conservación de Alimentos/métodos , Listeria/efectos de la radiación , Malus/microbiología , Leche/microbiologíaRESUMEN
The efficiency of UV-C irradiation as a non-thermal pasteurization process for liquid egg white (LEW) was investigated. LEW inoculated with Escherichia coli K-12 (ATCC 25253), pathogenic strain of Escherichia coli O157:H7 (NCTC12900) and Listeria innocua (NRRL B33314) were treated with UV light using a bench top collimated beam apparatus. Inoculated LEW samples were exposed to UV-C irradiation of known UV intensity of 1.314mW/cm(2) and sample depth of 0.153cm for 0, 3 5, 7, 10, 13, 17 and 20min. The populations of E. coli K-12, E. coli O157:H7 and L. innocua were reduced after 20min of exposure by 0.896, 1.403 and 0.960logCFU respectively. Additionally, the inactivation data obtained for each strain suspended in LEW was correlated by using Weibull (2 parameter), Log-Linear (1 parameter), Hom (2 parameter) and modified Chick Watson (2 parameter) models. The inactivation kinetics of E. coli K-12, E. coli O157:H7 and L. innocua were best described by modified Chick Watson model with the smallest root mean squared error (RMSE) (R(2)> or =0.92).
Asunto(s)
Clara de Huevo/microbiología , Irradiación de Alimentos , Conservación de Alimentos/métodos , Modelos Biológicos , Rayos Ultravioleta , Recuento de Colonia Microbiana , Seguridad de Productos para el Consumidor , Clara de Huevo/efectos de la radiación , Escherichia coli K12/crecimiento & desarrollo , Escherichia coli K12/efectos de la radiación , Escherichia coli O157/crecimiento & desarrollo , Escherichia coli O157/efectos de la radiación , Microbiología de Alimentos , Humanos , Cinética , Listeria/crecimiento & desarrollo , Listeria/efectos de la radiación , Factores de TiempoRESUMEN
The risk of listeriosis associated with ready-to-eat foods is a major concern in the United States. Pulsed light (PL) treatment has been effective for killing Listeria. The possibility of enhancing the antilisterial capability of PL treatment by combining PL with an additional hurdle, the natural antimicrobial nisin, was explored in this study. First, the ability of Listeria innocua to mimic the response of Listeria monocytogenes to PL treatment was demonstrated. Subsequently, a series of inoculation studies was performed in which canned sausages were surface inoculated with L. innocua as a surrogate for L. monocytogenes and then treated with a commercial preparation of nisin (Nisaplin), PL, or a combination of the two treatments. The application of a Nisaplin dip alone resulted in an immediate reduction of L. innocua by 2.35 +/- 0.09 log CFU. PL reduced L. innocua by 1.37 +/- 0.30 log CFU after exposure to 9.4 J/cm2. A total reduction of 4.03 +/- 0.15 log CFU was recorded after the combined treatment of Nisaplin and PL for 48 h at 4 degrees C. The long-term survival of L. innocua was evaluated on sausages stored at 4 degrees C. Treatment with Nisaplin and PL resulted in a 4- to 5-log reduction for two replicate studies. The combination treatment resulted in no significant microbial growth during 28 and 48 days of refrigerated storage in the first and second replicates, respectively. These results suggest that this combination treatment can be used as an effective antilisterial step in the production of ready-to-eat foods.