Effects of Pseudomonas chlororaphis and gaseous chlorine dioxide on the survival of Salmonella enterica on tomatoes

Control of Salmonella enterica on tomatoes is important for food safety. The aim of this research was to evaluate the survival of Salmonella enterica serovars Montevideo (SM) and Typhimurium (ST) on tomatoes exposed to gaseous chlorine dioxide and Pseudomonas chlororaphis (Pc). Pc was applied to stem scars of tomatoes prior to inoculations with SM and ST. Tomatoes were treated with gaseous ClO₂ at 0.4 mg L^?1 for 2 and 4 h (90% R.H. 13 °C), respectively. At 4 h of ClO₂ treatment, SM and ST populations were reduced to 0.82 and <0.30 log CFU g^?1, respectively. Tomatoes treated with SM and ST had 5.42 and 5.37 log CFU g^?1 of Salmonella. Tomatoes treated with Pc + Salmonella count was 2.59 (treated) and 5.83 log CFU g^?1 (control). Salmonella survival was similar at 2 and 4 h of ClO₂ treatment. Application of ClO₂ and Pc may reduce contamination of tomatoes by Salmonella serovars.     

Rapid Detection of Salmonella enterica Subspecies enterica serovar Typhimurium by Loop Mediated Isothermal Amplification (LAMP) Test From Field Chicken Meat Samples

Considering the importance of Salmonella enterica subsp. enterica serovar Typhimurium in the foodborne diseases, a Typhimurium specific loop mediated isothermal amplification (LAMP) test was standardized for its rapid detection in chicken meat. The Optimum results were obtained at 64^oC and 70 min temperature-time combination. The sensitivity of LAMP and PCR were compared with serial 10-fold dilution of the 100 ng of DNA. The LAMP test detected 2 pg DNA per reaction tube, whereas PCR detected 200 pg DNA per reaction. Therefore, the LAMP test was considered 100 times more sensitive than the PCR. The specificity of LAMP and PCR analyzed with six different isolates of non-Salmonella and 22 serovars of non-Typhimurium. None of these isolates were found positive by both LAMP and PCR. Twenty-eight pure isolates of Salmonella Typhimurium from diverse sources were also examined by Typhimurium specific LAMP and were all found positive. Two-hundred twenty-five field chicken meat samples were screened by cultural, PCR, and LAMP methods. The LAMP and PCR tests were performed by using DNA isolated from 8 h and 18 h enrichment samples, respectively. Typhimurium specific LAMP was shown to be in 100% correlated with cultural and PCR methods. However, LAMP test delivered the results within 26 h without sophisticated equipment while PCR and cultural methods took 48 h and 6 d, respectively. The LAMP test developed in this study has potential to detect and as well as differentiate S. Typhimurium from other Salmonella serovars.     

响应面法优化二氧化氯杀灭蛋壳大肠杆菌工艺研究

利用DesignExpert软件,根据Box-Behnken设计（BBD）原理采用四因素三水平的响应面分析法,对二氧化氯杀灭鲜蛋蛋壳大肠杆菌的条件进行了研究,获得了二次多元回归模型。对模型进行手动优化,利用优化模型优化出十组杀菌工艺参数,并对工艺参数进行了验证。     

Inactivation kinetics of inoculated Escherichia coli O157:H7 and Salmonella enterica on lettuce by chlorine dioxide gas

The purpose of this investigation was to study inactivation kinetics of inoculated Escherichia coli O157:H7 and Salmonella enterica on lettuce leaves by ClO(2) gas at different concentrations (0.5, 1.0, 1.5, 3.0, and 5.0 mg l(-1)) for 10 min and to determine the effect of ClO(2) gas on the quality and shelf life of lettuce during storage at 4 degrees C for 7 days. One hundred microliters of each targeted organism was separately spot-inoculated onto the surface (5 cm(2)) of lettuce (approximately 8-9 log CFU ml(-1)), air-dried, and treated with ClO(2) gas at 22 degrees C and 90-95% relative humidity for 10 min. Surviving bacterial populations on lettuce were determined using a membrane transferring method, which included a non-selective medium followed by a selective medium. The inactivation kinetics of E. coli O157:H7 and S. enterica was determined using first-order kinetics to establish D-values and z-values. The D-values of E. coli and S. enterica were 2.9+/-0.1 and 3.8+/-0.5 min, respectively, at 5.0 mg l(-1) ClO(2) gas. The z-values of E. coli and S. enterica were 16.2+/-2.4 and 21.4+/-0.5 mg l(-1), respectively. A 5 log CFU reduction (recommended by the United States Food and Drug Administration) for E. coli and S. enterica could be achieved with 5.0 mg l(-1) ClO(2) gas for 14.5 and 19.0 min, respectively. Treatment with ClO(2) gas significantly reduced inherent microflora on lettuce and microbial counts remained significantly (p<0.05) lower than the uninoculated control during storage at 4 degrees C for 7 days. However, treatment with ClO(2) gas had a significantly (p<0.05) negative impact on visual leaf quality. These results showed that treatment with ClO(2) gas significantly reduced selected pathogens and inherent microorganisms on lettuce; however, the processing conditions would likely need to be altered for consumer acceptance.     

Polyphenols content and antioxidant activity of Ghure (unripe grape) marc extract: influence of extraction time,temperature and solvent type

The objective of this study was to optimise the temperature (25–50 °C), time (1–24 h) and the solvent ratio (ethanol/water, 0–100%) for the phenolic compounds extraction of Ghure marc (unripe grape) using response surface methodology. The central composite design (CCD) generated satisfactory models for the optimisation of process variables. The results revealed that the ratio (X₃) was the most significant parameter on total phenolic content (TPC) and antioxidant activity [% 2‐diphenyl‐1‐picrylhydrazyl (DPPH)], and there was a significant correlation between TPC and antioxidant activity. The optimal conditions for temperature and time were 44.93 °C and 19.34 h, respectively, and the ratio of ethanol to water was 70.08, which verified with carrying out confirmatory experiments. Under this condition, TPC and %DPPH were 388.79 mg GAE per 100 g and 91.01, respectively. The results of this study revealed that the Ghure marc can be used as a low‐cost source of natural antioxidant in food.     

Inactivation kinetics of inoculated Escherichia coli O157:H7, Listeria monocytogenes and Salmonella enterica on strawberries by chlorine dioxide gas

Inactivation kinetics of inoculated Escherichia coli O157:H7, Listeria monocytogenes and Salmonella enterica on strawberries by chlorine dioxide gas at different concentrations (0.5, 1, 1.5, 3 and 5 mgl(-1)) for 10 min were studied. A cocktail of three strains of each targeted organism (100 microl) was spotted onto the surface of the strawberries (approximately 8-9 log ml(-1)) separately followed by air drying, and then treated with ClO(2) gas at 22 degrees C and 90-95% relative humidity. Approximately a 4.3-4.7 logCFU reduction per strawberry of all examined bacteria was achieved by treatment with 5 mgl(-1) ClO(2) for 10 min. The inactivation kinetics of E. coli O157:H7, L. monocytogenes and S. enterica were determined using first-order kinetic models to establish D-values and z-values. The D-values of E. coli, L. monocytogenes and S. enterica were 2.6+/-0.2, 2.3+/-0.2 and 2.7+/-0.7 min, respectively, at 5 mgl(-1) ClO(2). The z-values of E. coli, L. monocytogenes and S. enterica were 16.8+/-3.5, 15.8+/-3.5 and 23.3+/-3.3 mgl(-1), respectively. Furthermore, treatment with ClO(2) gas significantly (p < or = 0.05) reduced the initial microflora (mesophilic, psychrotrophic bacteria, yeasts and molds) on strawberries. Treatment with ClO(2) gas did not affect the color of strawberries and extended the shelf-life to 16 days compared to 8 days for the untreated control.     

Synergetic effect of petit-high pressure carbon dioxide combined with cinnamon (Cinnamomum cassia) essential oil against Salmonella typhimurium

This work aimed to evaluate the synergistic effect of petit-high pressure carbon dioxide (p-HPCD) and cinnamon (Cinnamomum cassia) essential oil (CEO) against Salmonella typhimurium. The results showed synergetic antibacterial activity of p-HPCD and CEO. After treatment with p-HPCD and CEO with a final concentration of 0.03125% under 1.3 MPa at 30 °C for 5 h, the population of S. typhimurium was decreased by 6.22 log₁₀ CFU/mL, significantly higher than p-HPCD or CEO alone. The combined treatment induced severe cellular changes in S. typhimurium, including cell membrane disruption, decreased intracellular protein and ATP, and intracellular reactive oxygen species (ROS) generation. Moreover, p-HPCD + CEO treatment significantly inhibited the expression of virulence genes including fliC, hilA, invI and ssrA, indicating that p-HPCD + CEO can reduce the production of virulence factors in S. typhimurium. This study provides a new approach to the application of p-HPCD and CEO in food processing and preservation.     

Effect of different packaging systems on the quality of tomato (Lycopersicon esculentum var. Rio Grande) fruits during storage

A packaging system is designed in our laboratory and evaluated for its suitability to extend storage life and improve the quality of tomato (var. Rio Grande) fruits at ambient temperature (Patent App. No. 484/2005). Freshly harvested mature green tomatoes were packed in polyethylene packaging with or without treating with calcium chloride, boric acid and potassium permanganate. The fruits were then evaluated for changes in quality parameters within the different stages of ripening. The treatments improved the storage life up to 96 days when compared with that of control (32 days). The results showed that within each ripening stage, the treated fruits remained better than that of control and all the ripening stages and treatments are significantly different ( P  < 0.05) from each other. It can also be deduced that treated fruits showed lower weight loss (%), total soluble solids (TSS) contents, acidity and spoilage incidence while ascorbic acid contents, sugar to acid ratio, flavour, texture, colour and overall acceptability were higher in treated fruits when compared with control at the red stage of ripening. Total sugars (%) were low in fruits treated with calcium chloride and boric acid with or without potassium permanganate.     

Effects of packaging materials following ethanol and benomyl treatments on chemical and microbiological changes in tomato (Lycopersicon esculentum) fruits

The effects of various post-harvest treatments on chemical changes, microbiological quality and occurrence of spoilage in ripe tomatoes (Lycopersicon esculentum L) were investigated. Fruits were pretreated in 70% ethanol or 0.2% benomyl before being packaged in low density polyethylene (LDPE), high density polyethylene (HDPE) or raffia palm basket. Fruit was stored at 27–32°C for up to 16 days. Control fruits (without pretreatment or packaging) showed the highest increase in pH; basket-packaged fruits were erratic but LDPE- and HDPE-packaged fruits showed gradual increases. Changes in titratable acidity were the inverse of those in pH. Among packagings, LDPE most effectively maintained total soluble solids (TSS) as compared with basket-packaged fruits, and the control showed the lowest TSS. Of the pretreatments, benomyl-treated fruits retained higher TSS. Aspergillus spp, Penicillium spp, Pseudomonas spp and Escherichia spp occurred most commonly in control and basket-packaged fruits, but Rhizopus spp and Leuconostoc spp predominantly occurred in LDPE- and HDPE-packaged fruits. Microbial load was lowest in benomyl-treated fruits followed by ethanol-treated and control fruits. Spoilage was higher (53.1%) in ethanol-treated than in benomyl-treated (43.8%) fruits.     

The influence of attachment to beef surfaces on the survival of cells of Salmonella enterica serovar Typhimurium DT104, at different a(w) values and at low storage temperatures

This study investigated the influence of attachment to beef surfaces on the survival, injury and death of stationary phase cells of Salmonella enterica serovar Typhimurium DT104, compared to cells free in solution. The effects on cells are considered at different a(w) values and low temperatures in relation to osmotic and cold temperature shock effects. Attachment of cells to meat surfaces prevented cell injury and death from hyperosmosis and low temperatures, compared to meat solutions. Storage of cells for 72h resulted in higher levels of cell death on cells attached to meat surfaces. The improved survival of cells in solutions was considered to be related to adaptation to osmotic stress as a result of exposure to a previous hyperosmotic shock and the ability of the cells to produce cold shock proteins. Pathogen cell growth at low temperatures is discussed in relation to the presence of low levels of NaCl. Finally the data is discussed in relation to pathogen survival on beef carcass surfaces during refrigeration.     

Optimisation of supercritical carbon dioxide extraction of lutein esters from marigold (Tagetes erect L.) with soybean oil as a co‐solvent

Response surface methodology was used to optimise the conditions of supercritical carbon dioxide extraction of lutein esters from marigold (Tagetes erect L.) with soybean oil as a co‐solvent. The effect of pressure, temperature, CO₂ flow rate and soybean oil concentration, and their interactions on the yield of lutein was investigated. Results showed that the data could be well fitted to a second‐order polynomial model with a R²‐value of 0.9398. The independent parameters of flow rate, pressure, temperature, all quadratics as well as the interactions between flow rate and pressure, and between flow rate and temperature affected the yield significantly (P ≤ 0.05). The model predicted that the optimal conditions were 35.5 MPa, 58.7 °C, CO₂ flow rate of 19.9 L/h with 6.9% of soybean oil as a co‐solvent, and under such conditions, the maximum yield of 1039.7 mg lutein /100 g marigold could be achieved.     

Optimisation of the combined treatments of aqueous chlorine dioxide,fumaric acid and ultraviolet‐C for improving the microbial quality and maintaining sensory quality of common buckwheat sprout

The effects of the combined treatments with aqueous chlorine dioxide (ClO₂), fumaric acid and ultraviolet‐C (UV‐C) on the microbial quality of common buckwheat sprouts were examined using a response surface methodology. The populations of total aerobic bacteria, yeast and mould, and coliform decreased with increasing aqueous ClO₂ and fumaric acid concentrations and increasing UV‐C irradiation dose. However, the increase in the UV‐C irradiation dose had a negative effect on the sensory quality. Therefore, the optimal combined treatment condition of 100 ppm aqueous ClO₂, 0.31% fumaric acid and 1.9 kJ m^?2 UV‐C was selected for the buckwheat sprouts by providing reductions of 3.9, 1.8 and 2.4 log CFU g^?1 on the populations of total aerobic bacteria, yeast and mould, and coliform, respectively. The combined treatment also maintained an acceptable sensory quality. These results suggest that the optimised combined treatment can be used as a microbial inactivation method for buckwheat sprouts.     

高密度CO_2处理提取鲵皮胶原蛋白的工艺优化

为研究高密度CO_2(dense phase carbon dioxide,DPCD)前处理对鲵皮胶原蛋白提取率的影响,以冻干鲵皮为原料,采用DPCD对鲵皮进行前处理,酶法提取鲵皮胶原蛋白,设计单因素试验和响应面试验,以胶原蛋白提取率为评价指标,优化DPCD处理鲵皮的工艺条件。结果表明,DPCD处理鲵皮的最适温度32℃、压强30 MPa、时间6 h;在该处理条件下,模型预测鲵皮胶原蛋白的提取率为39.70%,验证值为(39.48±0.74)%,无显著性差异(P0.05);与原料未经DPCD处理胶原蛋白提取率((20.63±0.46)%)相比,提取率提高了18.85%;微观结构显示,鲵皮经DPCD处理后的组织疏松、胶束纤维有规则排列被破坏并呈现多孔状态;电泳图谱显示鲵皮经DPCD处理后,胶原蛋白保持典型的Ⅰ型胶原蛋白结构特征。DPCD处理鲵皮,在保持鲵皮胶原蛋白结构特征的同时,显著提高了胶原蛋白提取率。     

Rhizosphere effect on survival of Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium in manure-amended soil during cabbage (Brassica oleracea) cultivation under tropical field conditions in Sub-Saharan Africa

The effect of cabbage (Brassica oleracea) rhizosphere on survival of Escherichia coli O157:H7 and Salmonella Typhimurium in manure-amended soils under tropical field conditions was investigated in the Central Agro-Ecological Zone of Uganda. Three-week old cabbage seedlings were transplanted and cultivated for 120 days on manure-amended soil inoculated with 4 or 7 log CFU/g non-virulent E. coli O157:H7 and S. Typhimurium. Cabbage rhizosphere did not affect survival of the 4 log CFU/g inocula in manure-amended soil and the two enteric bacteria were not detected on/in cabbage leaves at harvest. The 7 log CFU/g E. coli O157:H7 and S. Typhimurium survived in bulk soil for a maximum of 80 and 96 days, respectively, but the organisms remained culturable in cabbage rhizosphere up to the time of harvest. At 7 log CFU/g inoculum, E. coli O157:H7 and S. Typhimurium contamination on cabbage leaves occurred throughout the cultivation period. Leaf surface sterilisation with 1% AgNO₃ indicated that the organisms were present superficially and in protected locations on the leaves. These results demonstrate that under tropical field conditions, cabbage rhizosphere enhances the persistence of E. coli O157:H7 and S. Typhimurium in manure-amended soil at high inoculum density and is associated with long-term contamination of the leaves.