Biocontrol of Salmonella Typhimurium in RTE foods with the virulent bacteriophage FO1-E2

Antimicrobial agents can be incorporated into edible films to provide microbiological stability, since films can be used as carriers of a variety of additives to extend product shelf life and reduce the risk of microbial growth on food surfaces. Addition of antimicrobial agents to edible films offers advantages such as the use of small antimicrobial concentrations and low diffusion rates. The aim of this study was to evaluate inhibition by vapor contact of Aspergillus niger and Penicillium digitatum by selected concentrations of Mexican oregano (Lippia berlandieri Schauer), cinnamon (Cinnamomum verum) or lemongrass (Cymbopogon citratus) essential oils (EOs) added to amaranth, chitosan, or starch edible films. Essential oils were characterized by gas chromatography-mass spectrometry (GC/MS) analysis. Amaranth, chitosan and starch edible films were formulated with essential oil concentrations of 0.00, 0.25, 0.50, 0.75, 1.00, 2.00, or 4.00%. Antifungal activity was evaluated by determining the mold radial growth on agar media inoculated with A. niger and P. digitatum after exposure to vapors arising from essential oils added to amaranth, chitosan or starch films using the inverted lid technique. The modified Gompertz model adequately described mold growth curves (mean coefficient of determination 0.991 ± 0.05). Chitosan films exhibited better antifungal effectiveness (inhibition of A. niger with 0.25% of Mexican oregano and cinnamon EO; inhibition of P. digitatum with 0.50% EOs) than amaranth films (2.00 and 4.00% of cinnamon and Mexican oregano EO were needed to inhibit the studied molds, respectively). For chitosan and amaranth films a significant increase (p<0.05) of lag phase was observed among film concentrations while a significant decrease (p<0.05) of maximum specific growth was determined. Chitosan edible films incorporating Mexican oregano or cinnamon essential oil could improve the quality of foods by the action of the volatile compounds on surface growth of molds.     

壳聚糖/植物精油可食性抗菌膜研究进展

壳聚糖包埋植物精油制备的可食性抗菌膜具有原料来源广泛、可食、可降解,抗菌效果强等优势,在食品保鲜领域显示了重要的应用价值。本文综述了壳聚糖和精油各自的结构、性质、壳聚糖/精油复合膜及添加了其它天然高分子如蛋白质、淀粉等的复合可食性抗菌膜的研究进展,同时指出了目前该研究领域中存在的问题,并对未来的研究方向进行了展望,为壳聚糖/植物精油复合可食性抗菌膜研究的进一步开发利用提供参考。     

响应面试验优化玉米淀粉-壳聚糖可食膜的制备工艺

通过响应面法优化玉米淀粉、壳聚糖和甘油的质量分数来制备可食膜,以机械性能（伸长率、抗拉强度）和透湿性（water vapor permeability,WVP）为评价指标,得出二次响应预测模型。结果表明：玉米淀粉、壳聚糖和甘油的质量分数分别为3.71%、0.95%和0.64%时,抗拉强度最大;3 种物料质量分数分别为3.82%、0.50%和1.00%时,伸长率最大;3 种物料质量分数分别为3.52%、0.52%和0.50%时,WVP最小。综合考虑,玉米淀粉、壳聚糖和甘油质量分数分别为3.50%、0.50%和0.67%时,可食膜的性能最优。     

Antifungal activity evaluation of Mexican oregano (Lippia berlandieri Schauer) essential oil on the growth of Aspergillus flavus by gaseous contact

The antifungal activity of Mexican oregano (Lippia berlandieri Schauer) essential oil by gaseous contact on the growth of Aspergillus flavus at selected essential oil concentrations (14.7, 29.4, 58.8, or 117.6 μl of essential oil per liter of air) and temperatures (25, 30, or 35°C) was evaluated in potato dextrose agar formulated at water activity of 0.98 and pH 4.0. Mold growth curves were adequately fitted (0.984 < R(2) < 0.999) by the modified Gompertz model. The effect of the independent variables (concentration of essential oil and temperature) on the estimated model parameters (reciprocal of growth rate [1/ν(m)] and lag time [λ]) were evaluated through polynomial equations. Both ν(m) and λ were significantly (P < 0.05) affected by the independent variables; ν(m) decreased and λ increased as essential oil concentration increased and temperature decreased, which suggests that Mexican oregano essential oil retards or inhibits mold germination stage. Further, minimum fungistatic and fungicide essential oil concentrations at 30 and 35°C were determined. Mexican oregano essential oil applied in gas phase exerts important antifungal activity on the growth of A. flavus, suggesting its potential to inhibit other food spoilage molds.     

Antimicrobial Activity of Chitosan Films Enriched with Essential Oils

ABSTRACT: Antimicrobial and physicochemical properties of chitosan films and chitosan films enriched with essential oils (EO) were determined in vitro and on processed meat. Antimicrobial effects of pure EO of anise, basil, coriander, and oregano, and of chitosan-essential oil films against Listeria monocytogenes and Escheri-chia coli O157:H7 were determined by an agar diffusion test. The antibacterial effects of the EO were similar when applied alone or incorporated in the films. The intensity of antimicrobial efficacy was in the following order: oregano > > coriander > basil > anise. The chitosan films and chitosan-oregano EO films were applied on inoculated bologna samples and stored 5 d at 10 °C. Pure chitosan films reduced L. monocytogenes by 2 logs, whereas the films with 1% and 2% oregano EO decreased the numbers of L. monocytogenes by 3.6 to 4 logs and E. coli by 3 logs. Pure chitosan films were 89 μm thick, whereas addition of 1% and 2% oregano EO increased thickness to 220 and 318 μm, respectively. During application on bologna discs, the films absorbed moisture, resulting in the final thickness of 143, 242, and 333 μm, respectively. Addition of oregano essential oil into the chitosan films decreased water vapor permeability, puncture and tensile strength, but increased elasticity of the films. The films have the potential to be used as active biodegradable films with strong antimicrobial effects.     

Properties of Cassava Starch‐Based Edible Coating Containing Essential Oils

Edible coatings were produced using cassava starch (2% and 3% w/v) containing cinnamon bark (0.05% to 0.30% v/v) or fennel (0.05% to 0.30% v/v) essential oils. Edible cassava starch coating at 2% and 3% (w/v) containing or not containing 0.30% (v/v) of each essential oils conferred increased in water vapor resistance and decreased in the respiration rates of coated apple slices when compared with uncoated fruit. Cassava starch coatings (2% w/v) added 0.10% or 0.30% (v/v) fennel or cinnamon bark essential oils showed antioxidant capacity, and the addition of 0.30% (v/v) of each essential oil demonstrated antimicrobial properties. The coating containing cinnamon bark essential oil showed a significant antioxidant capacity, comparing to fennel essential oil. Antimicrobial tests showed that the addition of 0.30% (v/v) cinnamon bark essential oil to the edible coating inhibited the growth of Staphylococcus aureus and Salmonella choleraesuis, and 0.30% fennel essential oil inhibited just S. aureus. Treatment with 2% (w/v) of cassava starch containing 0.30% (v/v) of the cinnamon bark essential oil showed barrier properties, an antioxidant capacity and microbial inhibition.     

壳聚糖涂膜与牛至精油复配对鲜切菠萝的 保鲜作用

目的研究壳聚糖涂膜及与牛至精油复配对鲜切菠萝的保鲜的影响。方法在低温环境中,按照0.1%、0.3%和0.5%不同浓度的牛至精油与壳聚糖复配,并分别以水、壳聚糖涂膜处理为对照,分析不同处理方式对鲜切菠萝硬度、Vc含量、过氧化物酶(POD)、多酚氧化酶(PPO)、过氧化氢酶(CAT)和脂氧合酶(LOX)活性的影响。结果壳聚糖涂膜与0.5%的牛至精油复配后可减缓组织软化;与0.1%牛至精油复配可以维持Vc含量的缓慢降低。在不同处理组中,当含有0.3%和0.5%牛至精油时,鲜切菠萝的POD和CAT活性则呈下降趋势,而其他处理组则显著升高;当含有壳聚糖处理组时,其PPO和LOX活性则低于对照组。结论壳聚糖与牛至精油复配涂膜会在减缓鲜切菠萝营养损失的情况下延长其货架期。     

壳聚糖、溶菌酶和牛至油对大豆分离蛋白膜抑菌效果的影响

采用双倍稀释法研究壳聚糖、溶菌酶和牛至油对大肠杆菌和金黄色葡萄球菌的最小抑菌浓度。采用抑菌圈法研究添加天然抑菌剂壳聚糖、溶菌酶和牛至油的大豆分离蛋白膜对大肠杆菌、金黄色葡萄球菌、酿酒酵母和黑曲霉的抑菌效果。结果表明,添加壳聚糖、溶菌酶和牛至油的大豆分离蛋白膜对大肠杆菌、金黄色葡萄球菌、酿酒酵母和黑曲霉均有抑制作用。对大肠杆菌和金黄色葡萄球菌抑制效果为:牛至油>壳聚糖>溶菌酶。对酿酒酵母的抑制效果为:壳聚糖>牛至油>溶菌酶;对黑曲霉的抑制效果为:牛至油>壳聚糖>溶菌酶。因此,添加壳聚糖、溶菌酶和牛至油的大豆分离蛋白膜具有较好的抑菌效果和应用前景。     

Antimicrobial activity of whey protein based edible films incorporated with oregano,rosemary and garlic essential oils

The use of edible films to release antimicrobial constituents in food packaging is a form of active packaging. Antimicrobial properties of spice extracts are well known, however their application to edible films is limited. In this study, antimicrobial properties of whey protein isolate (WPI) films containing 1.0–4.0% (wt/vol) ratios of oregano, rosemary and garlic essential oils were tested against Escherichia coli O157:H7 (ATCC 35218), Staphylococcus aureus (ATCC 43300), Salmonella enteritidis (ATCC 13076), Listeria monocytogenes (NCTC 2167) and Lactobacillus plantarum (DSM 20174). Ten millilitres of molten hard agar was inoculated by 200 μl of bacterial cultures (colony count of 1 × 10⁸ CFU/ml) grown overnight in appropriate medium. Circular discs of WPI films containing spice extracts, prepared by casting method, were placed on a bacterial lawn. Zones of inhibition were measured after an incubation period. The film containing oregano essential oil was the most effective against these bacteria at 2% level than those containing garlic and rosemary extracts (P < 0.05). The use of rosemary essential oil incorporated into WPI films did not exhibit any antimicrobial activity whereas inhibitory effect of WPI film containing garlic essential oil was observed only at 3% and 4% level (P < 0.05). The results of this study suggested that the antimicrobial activity of some spice extracts were expressed in a WPI based edible film.     

Antibacterial Effects of Allspice, Garlic, and Oregano Essential Oils in Tomato Films Determined by Overlay and Vapor-Phase Methods

ABSTRACT:  Physical properties as well as antimicrobial activities against  Escherichia coli  O157:H7,  Salmonella enterica , and  Listeria monocytogenes  of allspice, garlic, and oregano essential oils (EOs) in tomato puree film-forming solutions (TPFFS) formulated into edible films at 0.5% to 3% (w/w) concentrations were investigated in this study. Antimicrobial activities were determined by 2 independent methods: overlay of the film on top of the bacteria and vapor-phase diffusion of the antimicrobial from the film to the bacteria. The results indicate that the antimicrobial activities against the 3 pathogens were in the following order: oregano oil > allspice oil > garlic oil.  Listeria monocytogenes  was less resistant to EO vapors, while  E. coli  O157:H7 was more resistant to EOs as determined by both overlay and vapor-phase diffusion tests. The presence of plant EO antimicrobials reduced the viscosity of TPFFS at the higher shear rates, but did not affect water vapor permeability of films. EOs increased elongation and darkened the color of films. The results of the present study show that the 3 plant-derived EOs can be used to prepare tomato-based antimicrobial edible films with good physical properties for food applications by both direct contact and indirectly by vapors emanating from the films.     

Antibacterial activity against E. coli O157:H7, physical properties, and storage stability of novel carvacrol-containing edible tomato films

Edible films containing plant antimicrobials are gaining importance as potential treatment to extend product shelf life and reduce risk of pathogen growth on contaminated food surfaces. The main objective of the present study was to evaluate the antimicrobial activities, storage stabilities, and physical-chemical-mechanica1 properties of novel edible films made from tomatoes containing carvacrol, the main constituent of oregano oil. The antimicrobial activities against E. coli O157:H7 and the stability of carvacrol were evaluated during the preparation and storage of tomato-based films made by 2 different casting methods, continuous casting and batch casting. Antimicrobial assays of tomato films indicated that optimum antimicrobial effects occurred with carvacrol levels of approximately 0.75% added to tomato purees before film preparation. HPLC analysis of the films indicated that the carvacrol concentrations and bactericidal effect of the films remained unchanged over the storage period of up to 98 d at 5 and 25 degrees C. Carvacrol addition to the tomato puree used to prepare the films increased water vapor permeability of tomato films. The continuous method for casting of the films appears more suitable for large-scale use than the batch method. This 1st report on tomato-based edible antimicrobial tomato films suggests that these films have the potential to prevent adverse effects of contaminated food and promote human health associated with the consumption of tomatoes.     

肉桂精油-玉米淀粉基抗菌膜的制备及其性能

为研究新型抗菌降解包装材料,筛选肉桂精油等4?种植物精油,以玉米淀粉、壳聚糖和魔芋葡甘露聚糖为成膜基质,甘油为增塑剂,吐温-80为表面活性剂,研究肉桂精油添加对复合膜机械性能、光学性能、阻水性能和抑菌性能的影响。结果表明：4?种精油对金黄色葡萄球菌、大肠杆菌和沙门菌的抗菌活性依次为肉桂精油＞牛至精油＞百里香精油＞迷迭香精油。随着肉桂精油质量浓度增加,复合膜的抗拉强度和水蒸气透过系数降低,断裂伸长率和不透明度升高。当肉桂精油质量浓度在15.0～20.0?g/L时,复合膜色泽指数a*值无明显差异（P＞0.05）,L*值显著降低,b*值和ΔE值显著增加（P＜0.05）。添加肉桂精油显著提高了玉米淀粉基膜的抗菌能力（P＜0.05）,精油与吐温-80相互作用对革兰氏阴性的大肠杆菌具有协同作用,而对革兰氏阳性的金黄色葡萄球菌具有拮抗作用。当肉桂精油质量浓度为20.0?g/L时,膜具有较好的物理性能和抗菌效果。本研究可为肉桂精油-玉米淀粉基可降解抗菌膜生产工艺参数的进一步优化提供参考。     

ANTIFUNGAL ACTIVITIES OF THYME, CLOVE AND OREGANO ESSENTIAL OILS

The antifungal potential of essential oils of oregano (Origanum vulgare), thyme (Thymus vulgaris) and clove (Syzygium aromaticum) was determined. To establish this antifungal potential, two molds related to food spoilage, Aspergillus niger and Aspergillus flavus, were selected. The agar dilution method was employed for the determination of antifungal activities. The three essential oils analyzed presented inhibitory effects on both molds tested. Oregano essential oil showed the highest inhibition of mold growth, followed by clove and thyme. Aspergillus flavus was more sensitive to thyme essential oil than A. niger. Clove essential oil was a stronger inhibitor against A. niger than against A. flavus.     

Antifungal Effect of Mexican Oregano (Lippia berlandieri Schauer) Essential Oil on a Wheat Flour-Based Medium

The antimicrobial activity of oregano has been attributed mainly to the presence of volatile compounds found in its essential oil (EO), mainly carvacrol and thymol. The search for antimicrobial activity of oregano EO with different concentrations of thymol and carvacrol, can lead to products with a wider range of applications. The aim of this work was to describe the in vitro antifungal effect of Mexican oregano (Lippia berlandieri Schauer) EO fractions on the growth of Aspergillus, Penicillium, and Rhizopus sp. The Mexican oregano EO fractions studied had different concentrations of carvacrol, which decreased from fraction 1 to 5 (81% to 23%), while thymol content increased from 3% to 64%. Fungal inhibition was evaluated on a wheat flour-based medium with EO fractions concentrations ranging from 50 to 200 mg/kg. Radial growth curves were fitted using the modified Gompertz model (R(2) (adj) = 0.989 ± 0.01). No significant differences (P > 0.05) were found with the different composition of the Mexican oregano EO fractions; nevertheless, fraction concentration presented significant (P < 0.05) mold inhibition as concentration increased. Rhizopus sp. (Rh18) showed a linear reduction on specific growth rate, on the maximum mold growth at the stationary phase, and an increase in the lag time as the concentration of the oregano EO increased; mold growth inhibition were achieved at 150 mg/kg in fractions 1 to 4, and at 100 mg/kg for fraction 5. Aspergillus sp. (As6) and Penicillium sp. (Pe36) were inhibited at 150 and 200 mg/kg, respectively. Results obtained suggest that Mexican oregano EO (Lippia berlandieri Schauer) compounds could be used as antimicrobial agents to prevent fungal growth in bakery products.     

Characterization of salmon gelatin based film on antimicrobial properties of chitosan against E. coli

Salmon gelatin and chitosan are high potential biopolymer to obtain edible films with antimicrobial effect for fresh meat. Therefore, it is important to characterize the structural (glassy or rubbery) state of the gelatin film on antimicrobial properties of chitosan. Extracted salmon (Salmo Salar) gelatin (acid-basic extraction) and low molecular weight chitosan solutions (0; 0.25; 0.5 and 1% w/w) at pH 5.5 were prepared to obtain a final concentration to 7% w/w (gelatin + chitosan). Films were obtained by casting at 5 °C and equilibrated at 33% and 85% of relative humidity (RH) in order to obtain a glassy and rubbery state, respectively. Water content, specific volume and glass transition temperature (Tg) were measured and antimicrobial properties against Escherichia coli (E.c.) (105 ufc/ml) were obtained by agar plate diffusion and kinetic measurements at 37 °C. Chitosan diffusion in agar plate was determined using liquid solutions and films over agar plate. The results showed diminution of Tg (maintaining structural state) as increase chitosan concentration, increasing also specific volume and water content due their high capacity to adsorb water. Although inhibition by chitosan of bacteria growth kinetic was 100%, solutions and films samples could not diffuse in the agar. In conclusion, the chitosan could not diffuse on the films matrix and also in agar plate and therefore in food when is combined with salmon gelatin. The implication of this work have been investigate the use of edible films from marine sources with antimicrobial effects in an effort to growing demand from consumers for safer and better quality foods.     

牛至精油羧甲基纤维素活性包装膜制备及其抗氧化和抗菌性能研究

本文利用牛至精油与羧甲基纤维素混合,制备具有抗氧化和抗菌性能的可降解活性包装膜。通过测定其厚度、颜色、机械性能和微观结构等,探究含有0%、1%、2%、3%牛至精油膜的结构和性能。结果表明:精油的加入一定程度增加了膜样的厚度,使膜样表面呈现较浅的棕黄色。精油与羧甲基纤维素相结合,膜样断裂伸长率随精油添加量增加无显著变化,抗拉强度显著降低(p>0.05)。微观结果显示,膜样表面出现微孔,使精油能够从中释放从而发挥抗氧化和抑菌作用。红外光谱的结果显示,牛至精油的加入没有与羧甲基纤维素发生化学反应。通过测定DPPH自由基清除率发现,牛至精油的加入能够显著提高膜的抗氧化能力,并随着精油浓度的提高而增强,但浓度为2%与3%差异并不显著(p>0.05)。利用琼脂扩散法发现,精油能够显著抑制金黄色葡萄球菌和大肠杆菌的生长,抑菌效果随着精油加入量的提升而显著提升(p<0.05)。综合膜样的性能和经济角度考虑得出,2%牛至精油含量的羧甲基纤维素膜具有良好的物理性能和较强的抗氧化和抑菌性能。     

Grapefruit Seed Extract and Lemon Essential Oil as Active Agents in Corn Starch–Chitosan Blend Films

The effect of lemon essential oil (LEO) and grapefruit seed extract (GSE) addition to starch–chitosan blend films on their functional properties and the associated structural changes were studied. Likewise, the films’ antioxidant properties and antimicrobial activity against some molds and Gram (+) and Gram (?) bacteria was evaluated. Composite active films exhibited a yellowish color, especially in the case of LEO addition. Film microstructure was analyzed by SEM evidencing a good incorporation of GSE to film matrix, while LEO droplets were dispersed in the polymer blend. Oxygen and water vapor barrier properties were not notably modified by the presence of active agents. FTIR analyses revealed that hydrogen bonding occurs in the blend films as the main interaction mechanism between components. Films containing LEO or GSE were less stiff and resistant but more stretchable than the control ones, being this effect concentration dependent. No notable antimicrobial action was observed in the films, which suggest that the required final concentration of active compounds must be fitted considering the minimum inhibitory concentration (MIC) values for a specific microorganism and its release kinetics to the food matrix.     

Inhibition of heat resistant molds: Aspergillus fumigatus and Paecilomyces variotii by some plant essential oils

In this study, the inhibitory properties of some essential oils including citrus (Citrus sinensis L. Osbeck), laurel (Laurus nobilis L.), myrtle (Myrtus communis L.), oregano (Origanum vulgare L.), and savory (Satureja thymbra L.) were investigated against the heat resistant molds Aspergillus fumigatus and Paecilomyces variotii isolated from margarine in a previous study in order to assess the potential for using these essential oils as a natural food preservative. In this study, the essential oils of the plants were obtained by steam distillation using Clevenger apparatus and were tested for antifungal activities at 0.25, 0.50, and 1.00% concentrations. Inhibitory effects of the essential oils on the growth of the fungi followed the sequence: oregano=citrus>savory>laurel>myrtle. P. variotii was more resistant against the essential oils than A. fumigatus.     

Antimicrobial activity of whey protein isolate edible films with essential oils against food spoilers and foodborne pathogens

The use of antimicrobial edible film is proposed as a means of improving food safety and extending the shelf-life of food systems by controlling the release of antimicrobials on food surfaces. In this work we first selected and studied 8 different essential oils (EOs) from plants, namely, oregano, clove, tea tree, coriander, mastic thyme, laurel, rosemary, and sage as natural antimicrobials against 2 gram-positive bacteria (Listeria innocua and Staphylococcus aureus) and 2 gram-negative bacteria (Salmonella enteritidis and Pseudomona fragi) by using the agar disk diffusion method. EOs from oregano, clove, and tea tree produced the largest surfaces of inhibition against the growth of the 4 bacterial strains tested. Second and following the assessment of compatibility, stable antimicrobial edible films based on whey protein isolate (WPI) with increasing concentrations (0.5% to 9%) of the 8 EOs were developed and tested for antimicrobial activity against the same gram-positive and gram-negative bacteria. WPI-edible films incorporating oregano or clove EO were found to have the most intense inhibitory effect of microbial growth. The bacterial strain gram-negative P. fragi presented the less susceptibility to the effect of those films. Moreover, only the edible films based on these 2 EOs were active against all 4 studied microorganisms. On the other hand, the edible films incorporating tea tree, coriander, mastic thyme, laurel, rosemary, or sage EOs even at high concentrations (7% to 9%) did not cause any antimicrobial effect against the pathogens S. aureus or S. enteritidis. PRACTICAL APPLICATION: Potential applications of this technology can introduce direct benefits to the food industry by improving safety and microbial product quality. The results of this research have direct application in the food industry with potential applications in various foodstuffs, including meat and poultry products where the control of spoilage bacteria such as P. fragi throughout their chilled storage or the improvement of food safety by controlling pathogens such as S. enteritidis are topics of particular interest for the industry.     

含精油可食性抗菌膜研究进展

可食性抗菌膜是目前食品包装领域研究的重要方向,含精油可食性抗菌膜是其研究的热点之一。本文介绍含精油的可食性食品包装膜中精油的主要成分及其抗菌特性,概述此类膜的一般制备技术,对含精油膜抗菌的抗菌性能测试方法进行归纳,就精油对可食性膜抗氧化性、机械性能、渗透性及其他理化性能的影响作重点阐述,最后分析含精油可食性抗菌膜可能存在的问题,并预测其发展前景。