Resistance of rubberwood (<Emphasis Type="Italic">Hevea brasiliensis</Emphasis>) treated with chitosan or silane against surface molds

The aim of this study was to determine the efficacy of chitosan and methoxysilane in the prevention of surface mold growth on rubberwood. Three different chitosan samples were tested; C1 (Mw 37 kDa), C2 (Mw 5.4 kDa) and C3 (Mw 3.5 kDa). Radial growth inhibition assay of the chitosan samples was investigated at concentrations ranging from 0.063 to 0.5 %w/v against Aspergillus niger BAM 4 and Penicillium decumbens CBS 121928. Chitosan samples C1 and C3 exhibited strong antifungal activity against both molds. Rubberwood samples were either vacuum or dip treated with varying concentrations of chitosan or silane solution. The content of chitosan in wood showed that after the leaching test, chitosan was well retained in both vacuum and dip treated wood. The concentration of silicon in wood showed similar results. The vacuum treated wood samples with chitosan C1 and C3 at 1 %w/v concentration had strong resistance against A. niger BAM 4. However, dip treated rubberwood samples with 2 %w/v chitosan solutions showed lower resistance against A. niger BAM 4. On the other hand, both vacuum and dip treated rubberwood samples with chitosan had no resistance against P. decumbens CBS 121928. The silane treated wood samples showed no resistance to fungal growth.     

Design of Sonotrode Ultrasound-Assisted Extraction of Phenolic Compounds from <Emphasis Type="Italic">Psidium guajava</Emphasis> L. Leaves

Psidium guajava L. has gained a special attention as health plant due to the presence of phenolic compounds. Box-Behnken design (BBD) has been applied for the extraction of target compounds from guava leaves via sonotrode ultrasound-assisted extraction (UAE). Different extraction times (5, 30, and 55 min), ratios of ethanol/water (50, 75, and 100% (v/v)), and ultrasound (US) power (80, 240, and 400 W) were tested to find their effect on the sum of phenolic compound (SPC), flavonols and flavan-3-ols via HPLC-ESI-QqQ-MS, and antioxidant activity (DPPH and TEAC assays). The best process conditions were as follows: 40 min, 60% ethanol/water (v/v), and 200 W. Established method has been used to extract phenolic compounds in two guava leaves varieties (pyrifera and pomifera). Pyrifera var. showed greater values of the SPC via HPLC-ESI-QqQ-MS (49.7 mg/g leaf dry weight (d.w.)), flavonols (12.51 mg/g d.w.), flavan-3-ols (7.20 mg/g d.w.), individual phenolic compounds, and antioxidant activity (8970 ± 5 and 465 ± 6 μmol Trolox/g leaf d.w, respectively) than pomifera var. Conventional extraction showed lower amounts of phenolic compounds (7.81 ± 0.03 and 4.64 ± 0.01 mg/g leaf d.w. for flavonols and flavan-3ols, respectively) in comparison to the ultrasound-assisted ones.     

Antioxidant Activity and Determination of Phenolic Compounds from <Emphasis Type="Italic">Eugenia involucrata</Emphasis> DC. Fruits by UHPLC-MS/MS

Fruits have been the focus of several studies aimed at finding new antioxidant sources for protection against the damage caused by reactive species. In this study, the antioxidant activity and the presence of phenolic compounds in all parts (peel, pulp, and seeds) of Eugenia involucrata DC. fruits were evaluated. DPPH^·, ABTS^·+, and ORAC methods were used to determine the antioxidant activity, and an UHPLC-MS/MS method was developed for determining the phenolic compounds (gallic, chlorogenic, ferulic, p-coumaric and ellagic acids, quercetin, and myricetin). In the determination of both antioxidant activity and phenolic composition, the efficiency of solvents with different polarities—methanol/H₂O (80:20, v/v), ethanol/H₂O (80:20, v/v), methanol/acidified water with phosphoric acid pH 3.00 (80:20, v/v), and ethyl acetate—for the extraction of the phenolic compounds, was also evaluated. All parts of E. involucrata fruits showed antioxidant activity, in the range of 36.68 ± 1.44 to 873.87 ± 18.24 μmol TE g^?1, being the highest values found in the seeds and peel when more polar extraction solvents were used. Six, five, and three phenolic compounds were identified and quantified in the pulp, peel, and seeds, respectively, with the highest abundance as p-coumaric acid (14 ± 2 mg kg^?1) in the pulp, quercetin (47 ± 5 mg kg^?1) in the peel, and gallic acid (74 ± 4 mg kg^?1) in the seeds, also when more polar solvents were used. Although antioxidant activity methods suggested that the peel and seeds have more antioxidant potential, a wider variety of compounds were determined in the pulp.     

Effects of Essential Oil Vapour Treatment on the Postharvest Disease Control and Different Defence Responses in Two Mango (<Emphasis Type="Italic">Mangifera indica</Emphasis> L.) Cultivars

The purpose of this study was to investigate the inhibitory effect of essential oils (thyme, clove and cinnamon) in vapour phase against the major fungal diseases of mango in vitro and in vivo. Thyme oil vapour (5 μL/Petri plate) completely inhibited the mycelial growth of Colletotrichum gloeosporioides and Lasiodiplodia theobromae under in vitro condition. Thyme oil vapour at 66.7 μL L^?1 significantly reduced artificially inoculated C. gloeosporioides and L. theobromae in mangoes for 4 days. GC/MS analysis revealed thymol, eugenol and benzofuran, 3-methyl as the dominant compounds in thyme, clove and cinnamon oils, respectively. The activities of defence and antioxidant enzymes including peroxidase, chitinase, phenylalanine ammonia-lyase, β-1,3-glucanase, catalase and superoxide dismutase were enhanced by thyme oil (66.7 μL L^?1) treatment and also help to maintain the phenolic content. Hence, postharvest thyme oil vapour treatment may prove to be an alternative means of controlling disease in mangoes.     

LC-ESI-MS/MS Analysis and Extraction Method of Phenolic Acids from Gluten-Free Precooked Buckwheat Pasta

In many countries, common buckwheat (Fagopyrum esculentum Möench) has been cultivated for its nutritive value as food ingredient. This plant is a rich source of vitamins and exogenic amino acids. Many of the health-promoting effects of F. esculentum have been attributed to a large amount of phenolic compounds. Presented in this paper, precooked buckwheat pasta, produced by extrusion cooking, is a gluten-free product without any technological additives. Moreover, it contains natural polyphenolic antioxidants and therefore could be classified as convenience food. The phenolic acid compositions of precooked buckwheat pasta were as follows: gallic, protocatechuic, gentisic, 4-OH-benzoic, vanillic, trans-caffeic, cis-caffeic, trans-p-coumaric, cis-p-coumaric, trans-ferulic, cis-ferulic, and salicylic. A very important step of sample pretreatment before quantitative analysis is the choice of extraction conditions. Therefore, in this study, before quantitative analysis (liquid chromatography-mass spectrometry, LC-ESI-MS/MS), optimization of ultrasound-assisted extraction (UAE) of phenolic acids from precooked buckwheat pasta was performed. The most effective conditions for the isolation of phenolic acids from precooked buckwheat pasta with the use of UAE were as follows: 80 % aqueous ethanol, 60 °C, ultrasound frequency 20 kHz, power 100 W, and time 40 min.     

Steady- and Unsteady-State Determination of the Water Vapor Permeance (<Emphasis Type="Italic">WVP</Emphasis>) of Polyethylene Film to Estimate the Moisture Gain of Packed Dry Mango

The water vapor permeance (WVP; g m^?2 d^?1 Pa^?1) of packaging films quantifying the water vapor transfer rate between foods and its surroundings is usually determined in units operating under steady-state conditions that do not necessarily reflect food handling scenarios. This study evaluated the determination of the WVP of a polyethylene (PE) film by steady-state method ASTM F1249-06 using a permeability cell and unsteady-state method ASTM E96/E96M in which 102 vacuum-sealed PE bags containing silica gel were stored (37.8 °C, 75% relative humidity) and weighed over 25 days. Average steady-state WVP (2.935 ± 0.365 × 10^?3, n = 4) fell within the 95% quantiles of unsteady-state WVP values (1.818–3.183 × 10^?3, n = 2142). Moisture uptake of dehydrated mango stored at 37.8 °C and 75% relative humidity was predicted with WVP values obtained by both methods. Predictions were validated by monitoring over 25 days the weight gain of 100 PE bags with dry mango. Experimental moisture averages during storage fell within one standard deviation of predictions using the unsteady-state WVP (R ² = 0.974). The same was observed only until day 15 for predictions obtained with the steady-state WVP. Calculations for days 20–25 overestimated the moisture uptake by 6.0–7.2%, resulting in registered R ² = 0.924. The unsteady-state WVP determination is low-cost, uses large numbers of film samples, and allowed more accurate predictions of dry mango moisture uptake. Knowledge of the moisture uptake controlled by the film WVP is essential when predicting the safety and quality changes limiting the shelf-life of foods.     

Screening of the phenolic profile and their antioxidative activities of methanol extracts of <Emphasis Type="Italic">Myrica rubra</Emphasis> fruits,leaves and bark

The phenolic composition in dried Myrica rubra fruits, leaves and bark were investigated for evaluation of its contribution to the antioxidant activity. The fruits, leaves and bark have the abundant phenolic compounds with the total phenolic content of 0.673, 0.276 and 0.136 mg/g (GA equivalents/FW), respectively. Ten phenolic compounds were isolated and identified in methanol extracts of Myrica rubra fruits by GC–MS analysis. Less phenolic compounds were found in leaves and bark than in fruits. However, the leaves and bark contain much higher concentrations of the trans-resveratrol over 100 μg/g than in fruits. The total antioxidant activities against the ·DPPH radical of those three samples were 0.438, 0.184 and 0.092 mg/g (Trolox equivalents/FW), respectively. The quantitative results indicated that a good correlation between the total antioxidant activity, total phenolic contents, and abundance of individual phenolic compound in Myrica rubra plants.     

Effect of Chitosan Film Coating Combined with Orange Peel Essential Oil on the Shelf Life of Deepwater Pink Shrimp

In this study, effect of chitosan films combined with orange (Citrus sinensis (L.) Osbeck) peel essential oil on the shelf life of deepwater pink shrimp (Parapenaeus longirostris Lucas 1846) was aimed. Chitosan (CH) and 2% orange peel essential oil (OPEO) combinated chitosan (CH+OPEO) were used for preparing film forming solution. Thickness and microstructure of the films, nutritional composition, sensory and melanosis evaluation, chemical, physical, and microbiological analyses were performed periodically and shelf-life was performed during the storage period of 15 days. The combination of chitosan film with OPEO was effective in prolonging the shelf life of fresh shrimps to 15 days (CH+OPEO), whereas the only chitosan-coated group had a shelf life of 10 days (CH) and the samples packaged without chitosan film had a shelf life of 7 days (control). The results of the study suggested that edible chitosan coatings together with OPEO preserved the shrimps and maintained the shelf life throughout the refrigerated storage period.     

Optimization of ultrasonic-assisted extraction of phenolic compounds from <Emphasis Type="Italic">Justicia spicigera</Emphasis> leaves

A Box–Behnken design (Extraction-time, pulse-cycle, sonication-amplitude) was employed to extract phenolic compounds from Justicia spicigera leaves by ultrasonic-assisted extraction. The muicle leaves extracts were analyzed measuring total phenolic compounds and antioxidant capacity. According to response surface methodology the optimal conditions of ultrasonic-assisted extraction to obtain the highest soluble phenolic content were 2 min (extraction time) for 0.7 s (pulse cycle) at 55% of sonication amplitude. Under these optimal conditions, the total phenolic content was higher when was used ultrasonic-assisted extraction (54.02 mg/g) than stirring (46.46 mg/g) and thermal decoction (47.76 mg/g); however, the antioxidant capacity from J. spicigera extracts did not increase by ultrasonic-assisted extraction. The extracts or aqueous infusions from J. spicigera leaves are used for therapeutic proposes, therefore the ultrasonic-assisted extraction is a useful technology to improve the extraction of phytochemicals from J. spicigera leaves.     

Characterizing the pH-Dependent Release Kinetics of Food-Grade Spray Drying Encapsulated Iron Microcapsules for Food Fortification

Iron deficiency is the primary cause of many widespread nutritional diseases including anemia, pregnancy complications, and infant mortality. Release kinetics of iron premixes to be mixed with food items like salt, rice, and tea is a key research objective of many globally active iron fortification efforts. Iron release kinetics of microcapsules of two reverse-enteric coating materials (chitosan and Eudragit EPO) encapsulating various amounts of ferrous sulfate (10–40% of total other solids) were done at three pH values (1, 4, 7) for 2 hours. Chitosan and Eudragit microcapsules contained 2.8–5.3% (w/w) and 1.7–9.6% (w/w) iron, respectively, depicting higher iron loading capacity of Eudragit microcapsules. More than 90% iron was released from most samples within 30 min under stomach conditions (pH 1) and less than 15% iron was released in 2 h under ambient conditions (pH 7), showing suitability of both chitosan and Eudragit EPO as reverse-enteric coatings for iron encapsulation. In terms of reverse-enteric behavior (RE), Eudragit EPO (RE = 2.4) was found to be slightly better than chitosan, suggesting the use of fillers in future research. Higuchi model and Hixson-Crowell model were found to best fit the data, suggesting a transport phenomenon governed by both (a) the diffusion process through the coating material and (b) the dissolution phenomenon resulting in decrease in size of the capsules. Results from this study shall provide guidance for technology development aspects of various food fortification initiatives and an understanding of the iron release from these fortificants during the food preparation and digestion stages.     

Antioxidative Effect of Seaweed Extracts in Chilled Storage of Minced Atlantic Mackerel (<Emphasis Type="Italic">Scomber scombrus</Emphasis>): Effect on Lipid and Protein Oxidation

In this study, antioxidant activity of absolute ethanol, 50 % ethanol and water extracts of two species of seaweeds namely, Fucus serratus and Polysiphonia fucoides were evaluated for their ability to retard lipid and protein oxidation in minced mackerel. Mackerel mince added with 0.5 g/kg of extracts was prepared. For comparison, BHT at 0.2 g/kg and a control with no added extracts were also prepared. The samples were stored at 5 °C for 8 days, and sampling was done at time 0, 1, 2, 4, 5 and 8 days. The 50 % ethanolic extracts of P. fucoides were found to be very effective in retarding lipid and protein oxidation, as it resulted in low levels of peroxide value, volatiles and carbonyl compounds and protected against the loss of α-tocopherol and tryptophan residues. In spite of the higher phenolic content, the absolute ethanol extracts of both species showed a pro-oxidative tendency in minced mackerel. Water extract with lowest phenolic content showed no antioxidant effect in minced mackerel. In conclusion, the 50 % ethanolic extracts of P. fucoides can be a potential source of natural antioxidants, as these extracts have antioxidant activities similar to synthetic antioxidants such as BHT. However, the extent of protection offered by these extracts against protein oxidation was not clear and further studies are needed to understand the nature of the interaction between proteins and these extracts.     

Development and characterization of reconstituted hydrogel from <Emphasis Type="Italic">Aloe vera</Emphasis> (<Emphasis Type="Italic">Aloe barbadensis</Emphasis> Miller) powder

Structural and rheological characterization of reconstituted hydrogels developed from A. vera non-fibrous alcohol insoluble residue (NFAIR) powder using different methods [viz., shaking (S), heating-shaking (HS), and heating (H)] and concentrations (viz., 0.2–1.6 %, w/v) was carried out. Functional group distribution by FTIR spectroscopy and Congo red (CR) method revealed the presence of acetylated acemannan in A. vera powder. Dynamic oscillation studies of A. vera (NFAIR) fluids at all concentrations of 0.2–1.6 %, w/v, showed gel strength in the order of H > HS > S method. However, in H method, increase in concentration from 0.2 to 1.6 %, w/v showed the conformational transition from semi-diluted solution to weak gel nature. Rheological models described the effect of heating temperatures (HT); 30–90 °C, and times (Ht); 15–60 min on viscoelastic behavior in reconstituted A. vera fluids. The reconstituted A. vera hydrogel prepared with a concentration of 1.6 %, w/v using 50 °C (HT) and 30 min (Ht) condition showed a good agreement with the Power law (storage modulus, G′) and Weak gel model (complex modulus, G*) fitted data (R² > 0.94) resulting higher viscoelastic moduli intercepts; G′₀ (71.5 Pa s ^n′), G″₀ (33.5 Pa s ^n″), lower slopes; n′ (0.22), n″ (0.06), higher network strength (A _F , 121.3 Pa s^1/z ) and number of network (z, 5.3) values. The obtained results suggested that heating at 50 °C/30 min can develop aqueous weak gel networks of A. vera with enhanced gel strength which may be utilized as a novel gelling agent for wide variety of targeted applications in food and pharmaceutical sectors.     

Smart NMR Method of Measurement of Moisture Content of Vegetables During Microwave Vacuum Drying

Microwave drying is usually combined with vacuum environment in conjunction with hot air flow to draw the moisture rapidly. The moisture content of the vegetables undergoing drying is hard to measure online. This research designed a microwave vacuum drying (MVD)-low-field nuclear magnetic resonance (NMR) smart device and investigated the feasibility of NMR method for online measurement of state of moisture during MVD. The relation between the signal amplitude (A ₂) and the true moisture content (M ₁) of six kinds of vegetables (mushroom, carrot, potato, lotus, edamame, vegetable corn) was fitted to estimate if NMR can measure the M ₁ of vegetables directly. Results showed that A ₂ and M ₁ of different fresh vegetables had no single empirical mathematical model to fit. However, for each kind of these vegetables, the A ₂ and corresponding M ₁ in different MVD stages showed a significant linear relationship. The predicted moisture content (M ₂) of mushroom: M ₂ = 5.25351 × 10^?4 A ₂ ? 0.34042, R = 0.996; carrot: M ₂ = 5.78756 × 10^?4 A ₂ ? 0.14108, R = 0.998; potato: M ₂ = 3.10019 × 10^?4 A ₂ ? 0.10612, R = 0.991; lotus: M ₂ = 2.32415 × 10^?4 A ₂ ? 0.01573, R = 0.998; edamame: M ₂ = 3.13310 × 10^?4 A ₂ ? 0.4198, R = 0.996; vegetable corn: M ₂ = 1.69461 × 10^?4 A ₂ ? 0.09063, R = 0.995. The linear models between M ₂ and A ₂ were able to estimate the end point (M ₁ < 8%) of MVD with a high accuracy (P > 0.950).     

Extraction and in vitro antioxidant capacity evaluation of phenolic compounds from pigmented aromatic rice (<Emphasis Type="Italic">Oryzae sativa</Emphasis> L.) cultivars

The aroma generating volatile components profile and in vitro antioxidant capacities of different aromatic rice cultivars was determined by GC–MS analysis and in terms of DPPH scavenging activity, lipid peroxidation inhibition, phosphomolybdenum reduction and reducing power assay. The total phenolic content including both free and bound forms in the analyzed aromatic rice cultivars, Mushki budgi (1.62 mg GAE/g), Mushki kandi (1.63 mg GAE/g) and Kamad (1.60 mg GAE/g) were found double the amount as compared to non-aromatic Koshkari (0.86 mg GAE/g) cultivar. The aromatic rice cultivars had also shown higher total flavonoid content and antioxidant activity than non-aromatic rice cultivar (Koshkari). The GC–MS results indicated 21-aromatic compounds present in sufficient quantities in aromatic cultivars and some of them were unique to these cultivars. Among the compounds identified, aldehydes were found in higher quantity followed by alkanes, ketones and esters. Among the aromatic rice cultivars, Mushki budgi and Mushki kandi were found possessing higher quantity of flavoring components such as benzaldehyde, a carcinostatic agent. The cultivars Mushki budgi and Mushki kandi indicated positive correlation of TPC, TFC and the in vitro antioxidant components largely, while the less aromatic Kamad, correlate with only two components viz DPPH and lipid peroxidation.     

FAIMS based sensing of <Emphasis Type="Italic">Burkholderia cepacia</Emphasis> caused sour skin in onions under bulk storage condition

Onion (Allium cepa L.) is one of the most important vegetable crops in the Pacific Northwestern states of the U.S., and is stored for 9–12 months under bulk storage condition. Portable non-contact sensing technologies are needed to effectively detect pathogenic infections to the onion bulbs during bulk storage periods for real-time crop loss management. Therefore, portable field asymmetric ion mobility spectrometry (FAIMS) was evaluated towards detection of onion sour skin caused by Burkholderia cepacia, an important postharvest disease under bulk storage conditions. Studied were onions stored at 25?°C under two treatment conditions, inoculated with B. cepacia and with sterile water (as control), during 21-days temporal storage. Results confirmed applicability of FAIMS to detect volatile organic compounds pertinent to B. cepacia caused sour skin as early as 3 days after inoculation. Principal component analysis (PCA) score plots for FAIMS extracted data illustrated distinct clusters corresponding to inoculated and healthy treatments. PCA also suggested a significant range of dispersion field intensity (47–77%) and compensation voltage (?0.24 to 0.48 V), which could potentially be used to train portable FAIMS for real-time detection of sour skin under bulk storage conditions.     

Effects of chitosan and collagen containing α-tocopherol on the oxidative stability in bulk oil and oil-in-water emulsion

To provide efficient antioxidant capacities, proper carriers are needed to protect antioxidants against oxidative stress. Collagen mesh structure or chitosan gel was loaded with α-tocopherol and their effects were evaluated in bulk corn oil or oil-in-water (O/W) emulsion at 60 °C. Added collagen and chitosan enhanced oxidative stability in corn oil and O/W emulsions at 60 °C compared to corn oils without carriers or with addition of α-tocopherol (p < 0.05). Stability of α-tocopherol in corn oil loaded in collagen or chitosan was significantly enhanced compared to that in oils without carriers (p < 0.05). In O/W emulsions, α-tocopherol loaded collagen showed higher antioxidant properties than α-tocopherol loaded chitosan (p < 0.05). Collagen mesh structure and chitosan gel retarded the rates of lipid oxidation efficiently in both food matrices when α-tocopherol was not loaded. Collagen mesh structure and chitosan gel can be useful carriers for α-tocopherol in bulk oil or O/W emulsion.     

Effects of organic acid pretreatment on microstructure,functional and thermal properties of unripe banana flour

Starch availability has been implicated in unripe matured banana (Musa species), which when processed yields flour suitable for application in low gluten and composite wheat formulations. Unripe Musa species: Williams, Luvhele, Mabonde and Muomva-red obtained from fruit bunch were pretreated with ascorbic, citric and lactic acids, processed into 50 g of flour and characterised for their functional and thermal properties. Scanning electron microscope of unripe banana flour (UBF) showed varying micrographs of flour, with polygonal for Luvhele, oval for Mabonde, elongated for Muomva-red and between polygonal and spherical for Williams. The bulk density of UBF samples was within the range of 0.66–0.84 g/mL for all organic acid pretreatment while citric acid pretreated UBF had the least browning index. Significant difference (p < 0.05) was recorded in swelling power with no significant difference in water solubility index except for Mabonde UBF. Thermal properties showed single endothermic transition for all UBF samples at various pretreatment concentration. The onset temperature (To) of UBF ranges from 49.82 to 65.59 °C, peak temperature (Tp) from 60.11 to 76.71 °C, conclusion temperature (Tc) from 70.36 to 94.16 °C and enthalpy of gelatinization (ΔH) from 2.61 to 32.24 J/g. Short amylopectin chains present in starch of UBF was attributed to low To, Tp, Tc and ΔH values recorded for Mabonde cultivar, while the contribution of heat-moisture treatment rather than organic acid pretreatment of UBF samples was attributed to different gelatinization and transition temperatures recorded for all cultivars examined.     

Drying kinetics,colour, total phenolic content and antioxidant capacity properties of kiwi dried by different methods

This study analysed the convective (60, 70 and 80° C), microwave (120 and 350 W) and freeze drying methods in terms of their effects on the drying characteristics, colour, total phenolic content (TPC) and antioxidant capacity of kiwi slices. Nine different mathematical models were applied to experimental data to achieve the most accurate calculation for drying curves. The Midilli et al. and Wang and Singh models proved to be the most suitable at explaining the drying kinetics of kiwi samples as compared to other models according to the statistical tests. Each drying method was significantly affected by colour parameters (L*, a*, b*, C, α and ΔE). The dried samples exhibited respectively 5–49 % and 10–47 % less TPC and antioxidant capacity compared to the fresh sample. According to the correlation analysis conducted between TPC and antioxidant capacity for kiwi slices, there is a positive correlation (R ²  = 0.7796). Microwave dried samples at 120 W particularly had the lowest TPC and antioxidant capacity. Freeze drying method yielded the closest values with respect to colour values, total phenol content and antioxidant capacity to those of fresh samples when compared to the other methods.     

Enterohemorrhagic <Emphasis Type="Italic">Escherichia coli</Emphasis> (EHEC) in water from karst springs: detection with real-time PCR and isolation of strains

Within 2 months, two water sources in a karst area in Switzerland were sampled 9 times each, and analyzed by real-time PCR for 6 EHEC O-types, Shiga-like-toxin (stx1 and stx2) and intimin (eae) genes. With the exception of O111, 5 O-types were recorded regularly and at high frequencies (O26: 33.3 %; O157: 33.3 %; O104: 66.6 %; O103: 72.2 %; O145: 94.4 %). Genes for Shiga-like-toxins and intimin were almost omnipresent (stx1: 77.8 %; stx2: 83.3 %; eae: 77.8 %). Strain isolation was undertaken for O-groups 26, 103, 104, 145 and 157. Sample selection for strain isolation was based on Cq-values for the O-groups and stx1, stx2 and eae. From selected samples, frozen enrichment cultures were cultivated on EHLY-agar and 50 typical colonies screened for the O-type and genes encoding for stx1, stx2 and eae. With this approach, only one virulent EHEC-strain could be isolated (Escherichia coli O103, stx1 +; stx2 ?; eae +). We carried out one extensive testing with 800 colonies of O-group O145, and no virulent strain was isolated. Our findings showed that PCR-results are not sufficient to formulate epidemiological conclusions and that the isolation of strains is necessary. However, as the detection procedure of EHEC in foods is cumbersome and expensive, the appropriateness of such an approach in official food control is a matter of debate.     

Comparative Study on the Inactivation and Photoreactivation Response of <Emphasis Type="Italic">Listeria monocytogenes</Emphasis> Seafood Isolates and a <Emphasis Type="Italic">Listeria innocua</Emphasis> Surrogate after Pulsed Light Treatment

The inactivation and photoreactivation response of six seafood-isolated Listeria monocytogenes and one Listeria innocua strain after pulsed light (PL) treatment was evaluated. The lower inactivation levels found after exposure of treated samples to daylight during the first 90 min of storage confirmed that both L. innocua and L. monocytogenes have the capability to photorepair PL-induced DNA damage upon appropriate conditions. Photoreactivation levels from 0.2 to 2.1 log CFU cm^?2 were observed depending on treatment intensity (fluence) and Listeria strain. Complete photorepair of PL-caused damage was not found even after treatments inducing low inactivation levels. Photoreactivation increased up to 2.1 log with the applied fluence up to a threshold able to cause between 2.4 and 5.4 log reductions under dark storage. Photorepair was not avoided but lower photoreactivation was observed after higher fluence inducing more than 6 log reductions under dark storage. Both L. innocua and L. monocytogenes serotype 1/2b exhibited the highest photoreactivation levels whereas serotypes 1/2a showed the lowest ones. The overall inactivation and photoreactivation responses of tested Listeria strains were comparable indicating that L. innocua may be a good surrogate for the safe evaluation, optimization and validation of PL technology to control L. monocytogenes in food products and food processing facilities.