Mould and yeast flora in fresh berries, grapes and citrus fruits

Fresh fruits are prone to fungal contamination in the field, during harvest, transport, marketing, and with the consumer. It is important to identify fungal contaminants in fresh fruits because some moulds can grow and produce mycotoxins on these commodities while certain yeasts and moulds can cause infections or allergies. In this study, 251 fresh fruit samples including several varieties of grapes, strawberries, blueberries, raspberries, blackberries, and various citrus fruits were surface-disinfected, incubated at room temperature for up to 14 days without supplemental media, and subsequently examined for mould and yeast growth. The level of contamination (percent of contaminated items/sample) varied depending on the type of fruit. All raspberry and blackberry samples were contaminated at levels ranging from 33% to 100%, whereas 95% of the blueberry samples supported mould growth at levels between 10% and 100% of the tested berries, and 97% of strawberry samples showed fungal growth on 33-100% of tested berries. The most common moulds isolated from these commodities were Botrytis cinerea, Rhizopus (in strawberries), Alternaria, Penicillium, Cladosporium and Fusarium followed by yeasts, Trichoderma and Aureobasidium. Thirty-five percent of the grape samples tested were contaminated and supported fungal growth; the levels of contamination ranged from 9% to 80%. The most common fungi spoiling grapes were Alternaria, B. cinerea and Cladosporium. Eighty-three percent of the citrus fruit samples showed fungal growth at levels ranging from 25% to 100% of tested fruits. The most common fungi in citrus fruits were Alternaria, Cladosporium, Penicillium, Fusarium and yeasts. Less common were Trichoderma, Geotrichum and Rhizopus.     

进口鲜葡萄市场病害的检测与分析

对上海市场销售的进口葡萄进行了市场病害的检测,在原产于美国和智利的红地球葡萄（red-globe grape）中检出了链格孢（Alternaria spp）、灰霉（Botrytis cinerea）、芽枝霉（Cladosporium spp）和匐柄霉（Stemphulium sp）四个属的潜伏病原真菌,其中Alternaria spp在健康果实中的检出率高达65.8%,携带外来菌的可能性较大。同时进口红地球葡萄存在较严重的SO2伤害,在所检测的样品中有伤害症状的果粒占所检测果粒数的比率最高可达92%。     

Antifungal Coatings on Fresh Strawberries (Fragaria × ananassa) to Control Mold Growth During Cold Storage

ABSTRACT: Chitosan (2%) -based or hydroxypropyl methylcellulose (HPMC) (1%) -based coatings were applied on fresh strawberries to evaluate their antifungal efficacies against Cladosporium sp. and Rhizopus sp. Potassium sorbate (PS) was also incorporated into coating formulas to evaluate any additional inhibitory effects on mold inhibition. Strawberries were inoculated with Cladosporium sp. or Rhizopus sp. at a level of approximately 10³ log colony-forming units (CFU) /g, coated with 2% chitosan, 2% chitosan containing 0.3% PS, or 1% HPMC containing 0.3% PS, and stored at about 5°C and about 50% RH up to 23 d for enumeration of mold, yeast, total aerobes, and coli forms. To assess antifungal activity of the coating materials in vitro, coating solutions were embedded into agar plates and the diameters of radial mold growth were measured after inoculation. In addition, weight loss of coated strawberries and water vapor permeability of the coatings were measured. No significant combined inhibitory effects between chitosan and PS on fungal growth on fresh strawberries were detected. However, significant combined inhibition activity was observed in in vitro testing when PS was formulated into chitosan. Antifungal activity of chitosan against Cladosporium sp. and Rhizopus sp. was not affected by the autoclaving process of coating solutions. Coating treatment also reduced total aerobic count, coliforms, and weight loss of strawberries during storage. Hence, chitosan can be used as a natural antimicrobial coating on fresh strawberries to control the growth of fungi, thus extending shelf-life of the fruits.     

Production of alternariol and alternariol methyl ether by Alternaria alternata grown on fruits at various temperatures

Two toxigenic strains of the fungus Alternaria alternata (ATCC 56836 and ATCC 66868) were grown on surface-disinfected, fresh, ripe fruits and tested for the production of alternariol (AOH) and alternariol methyl ether (AME). Examined fruits included strawberries; red and green seedless grapes; concord grapes; red delicious, golden delicious, and gala apples; and blueberries. After inoculation, fruits were incubated at 4, 10 degrees C, or room temperature (approximately 21 degrees C) for up to 3 weeks. At weekly intervals, duplicate samples were analyzed for AOH and AME by using liquid chromatography. Results indicated that A. alternata and its metabolites were not a major problem in strawberries due to the presence of fast-growing molds like Rhizopus and Botrytis that outgrew and possibly inhibited Alternaria. Both Alternaria strains showed limited growth on apples, although fast-growing molds were not present after surface disinfection; AOH and AME were produced only by the ATCC 56836 strain on the golden delicious and gala varieties, (ranging from <0.1 to 5 microg/g and <0.1 to 14 microg/g for AOH and AME, respectively). Restricted growth of both strains without toxin production occurred in blueberries, whereas moderate growth and AOH (<0.1 to 3,336 microg/g) and AME (<0.1 to 1,716 microg/g) production took place in grapes.     

凉果中优势腐败霉菌的分离鉴定

凉果加工与贮藏中霉菌污染直接影响凉果的品质。为明确凉果贮藏中优势腐败霉菌的种类,该研究对在常温贮存4个月后霉变的葡萄干、话梅、杏脯中腐败霉菌进行了分离纯化,得到7种典型腐败霉菌。将分离出的腐败霉菌回接至无菌的葡萄干、话梅、杏脯表面,进行回接侵染实验,通过凉果霉变时间确定出4株菌株为优势腐败霉菌。通过菌落形态特征、菌丝和分生孢子显微结构观察,结合核糖体内转录间隔区（rDNA-ITS）序列分析,构建系统发育,鉴定优势霉菌种属。结果表明：葡萄干的优势霉菌为芽枝状枝孢霉（Cladosporium velox）和暗黄青霉（Penicillium citreonigrum）,话梅的优势腐败菌为细极链格孢（Alternaria tenuissima）,杏脯的优势腐败霉菌为菌核青霉（Penicillium sclerotiorum）。该研究结果可为凉果腐败霉菌的分离鉴定提供可靠的方法,也可为后续凉果杀菌方法的选择,贮藏条件的优化提供依据。     

Using survival analysis to investigate the effect of UV-C and heat treatment on storage rot of strawberry and sweet cherry

Ultraviolet light and heat treatment are proposed as alternative techniques for the use of chemicals to reduce the development of the spoilage fungi Botrytis cinerea and Monilinia fructigena on strawberry and sweet cherry, respectively, during storage. In order to investigate the effect of both physical techniques on microbial inactivation and on fruit quality, inoculated berries were subjected to different temperatures (40-48 degrees C) and UV-C doses (0.05-1.50 J/cm2). For each condition, 20 berries were used. After the treatment, fungal growth, visual damage (holes, stains) and fruit firmness were evaluated during a period of 10 days. The experimental data were analysed statistically using survival analysis techniques. Fungal growth on strawberries was significantly retarded using UV-C doses of 0.05 J/cm2 and higher. The same treatment had no significant effect when applied to cherries. The highest doses (1.00 and 1.50 J/cm2) had a negative effect on the calyx of the strawberry, causing browning and drying of the leaves. No beneficial effect of a low temperature treatment (40-48 degrees C) on the shelf life of strawberries was observed, but fungal development on cherries was retarded at temperatures of 45 and 48 degrees C. These temperatures caused severe damage on strawberries (soft stains, holes, decreased firmness), but had no influence on the quality of sweet cherries.     

河北省市售水果中植物生长调节剂残留状况 调查与分析

目的了解河北省市售水果(草莓、葡萄、猕猴桃、樱桃和西瓜)中植物生长调节剂赤霉素、氯吡脲、噻苯隆、2, 4-二氯苯氧乙酸、4-氯苯氧乙酸和多效唑残留状况。方法对河北地区148份草莓、122份葡萄、51份猕猴桃、39份樱桃和9份西瓜中植物生长调节剂残留状况进行研究。参考食品中化学污染物及有害因素监测技术手册方法,采用液相色谱-串联质谱法进行检测。结果 148份草莓样品中多效唑检出39份; 51份猕猴桃样品中氯吡脲检出23份; 39份樱桃样品中2,4-二氯苯氧乙酸检出16份;其他植物生长调节剂在各类水果中检出率均不高。结论植物生长调节剂在河北省市售的各类水果中存在不同程度的残留,但残留量较低,总体安全性较好。     

Fungal spoilage of bottled mineral water

The occurrence of filamentous fungi together with bacteriological parameters was assessed in 126 samples of still bottled mineral water of eight different commercial brands in Argentina. In spoiled samples with visible mycelium growth, the most frequently isolated fungal species were Penicillium citrinum, P. glabrum, other Penicillium species, Cladosporium cladosporioides and Alternaria alternata. In unspoiled samples, the genera found were Penicillium, Cladosporium, Rhizopus, Aspergillus and Phoma. Only three of the 126 samples failed to meet the required microbiological standards because they were found to contain faecal streptococci.     

Volatile Compounds from Raspberry and Strawberry Fruit Inhibit Postharvest Decay Fungi

Fifteen volatiles released by red raspberries and strawberries during ripening were analyzed for the inhibition of postharvest decay fungi. Five compounds completely inhibited all fungi directly on fruit at 0.4 μL/mL., Of these, benzaldehyde at 0.04 μL/mL completely inhibited isolated cultures of Alternaria altemata, Botrytis cinerea, and Col-Ietotrichum gloeosporioides, while 1-hexanol, E-2-hexenal and 2-non-anone inhibited all three fungal species at 0.1 μL/mL. When added directly to media, 0.1% (v/v) benzaldehyde or E-2-hexenal inhibited all three species. Starch encapsulated 2-nonanone reduced fungal decay of raspberries and strawberries in enclosed containers after 7 days at 10°C.     

猕猴桃和青菜病原真菌的分离、鉴定及生物学特性

目的:猕猴桃和青菜在储存和运输的过程中容易因病原菌导致腐烂而造成经济损失,因此,了解其病原菌的种类和特性对病原菌的防治十分必要。方法:本研究通过致病性鉴定、形态学鉴定及ITS鉴定从发病猕猴桃及青菜中分离得到病原真菌,并且对其进行了生物学特性的探究。结果:从发病猕猴桃中分离纯化得到4株病原真菌,经鉴定为壳囊孢属Cytospora sp.、波兰青霉Penicillium polonicum、加州灰霉菌Botrytis californica和盖姆斯木霉Trichoderma gamsii。从发病青菜中分离纯化得到3株病原真菌,经鉴定为圆形镰刀菌Fusarium circinatum、枝孢梭菌Cladosporium angustiterminale和加州灰霉菌Botrytis californica。壳囊孢属在10~28 ℃范围内均可生长,其余6株病原真菌在4~30 ℃范围内均可生长,并且最适生长温度为20~28 ℃。除枝孢梭菌外,其余6株病原真菌的的最适pH在6~8之间。枝孢梭菌的耐酸性较强,其最适pH可能在4以下。壳囊孢属在pH为4~9范围内均可以生长,其余6株病原真菌在pH为4~10范围内均可以生长。结论:经致病性鉴定后,发现这7株真菌均可对应地引起猕猴桃和青菜腐烂,而其中的加州灰霉菌Botrytis californica可同时引起猕猴桃和青菜的腐烂。波兰青霉、加州灰霉菌、盖姆斯木霉和枝孢梭菌在4 ℃情况下均可以生长,说明冰箱冷藏的温度并不能防止这些病原菌侵染蔬果。本文为猕猴桃及青菜病原真菌的防治提供了参考依据。     

Quantitative analysis of mycoflora on commercial domestic fruits in Japan

A comprehensive and quantitative analysis of the mycoflora on the surface of commercial fruit was performed. Nine kinds of fruits grown in Japan were tested. Overall fungal counts on the fruits ranged from 3.1 to 6.5 log CFU/g. The mean percentages of the total yeast counts were higher than those of molds in samples of apples, Japanese pears, and strawberries, ranging from 58.5 to 67.0%, and were lower than those of molds in samples of the other six fruits, ranging from 9.8 to 48.3%. Cladosporium was the most frequent fungus and was found in samples of all nine types of fruits, followed by Penicillium found in eight types of fruits. The fungi with the highest total counts in samples of the various fruits were Acremonium in cantaloupe melons (47.6% of the total fungal count), Aspergillus in grapes (32.2%), Aureobasidium in apples (21.3%), blueberries (63.6%), and peaches (33.6%), Cladosporium in strawberries (38.4%), Cryptococcus in Japanese pears (37.6%), Penicillium in mandarins (22.3%), and Sporobolomyces in lemons (26.9%). These results demonstrated that the mycoflora on the surfaces of these fruits mainly consists of common pre- and postharvest inhabitants of the plants or in the environment; fungi that produce mycotoxins or cause market diseases were not prominent in the mycoflora of healthy fruits. These findings suggest fruits should be handled carefully with consideration given to fungal contaminants, including nonpathogenic fungi, to control the quality of fruits and processed fruit products.     

In vitro binding of bile acids by blueberries (Vaccinium spp.), plums (Prunus spp.), prunes (Prunus spp.), strawberries (Fragaria X ananassa), cherries (Malpighia punicifolia), cranberries (Vaccinium macrocarpon) and apples (Malus sylvestris)

The in vitro binding of bile acids by blueberries (Vaccinium spp.), plums (Prunus spp.), prunes (Prunus spp.), strawberries (Fragaria X ananassa), cherries (Malpighia punicifolia) cranberries (Vaccinium macrocarpon) and apples (Malus sylvestris) was determined using a mixture of bile acids secreted in human bile at a duodenal physiological pH of 6.3. Six treatments and two blank incubations were conducted to testing various fresh raw fruits on an equal dry matter basis. Considering cholestyramine (bile acid binding, cholesterol lowering drug) as 100% bound, the relative in vitro bile acid binding on dry matter (DM), total dietary fiber (TDF) and total polysaccharides (PCH) basis was for blueberries 7%, 47% and 25%; plums 6%, 53% and 50%; prunes 5%, 50% and 14%; strawberries 5%, 23% and 15%; cherries 5%, 37% and 5%; cranberries 4%, 12% and 7%; and apple 1%, 7% and 5%, respectively. Bile acid binding on DM basis for blueberries was significantly (P ? 0.05) higher than all the fruits tested. The bile acid binding for plums was similar to that for prunes and strawberries and significantly higher than cherries, cranberries and apples. Binding values for cherries and cranberries were significantly higher than those for apples. These results point to the relative health promoting potential of blueberries > plums = prunes = strawberries = cherries = cranberries > apples as indicated by their bile acid binding on DM basis. The variability in bile acid binding between the fruits tested maybe related to their phytonutrients (antioxidants, polyphenols, hydroxycinnamic acids, flavonoids, anthocyanins, flavonols, proanthocyanidins, catechins), structure, hydrophobicity of undigested fractions, anionic or cationic nature of the metabolites produced during digestion or their interaction with active binding sites. Inclusion of blueberries, plums, prunes, strawberries, cherries and cranberries in our daily diet as health promoting fruits should be encouraged. Animal studies are planned to validate in vitro bile acid binding of fruits observed herein to their potential of atherosclerosis amelioration (lipid and lipoprotein lowering) and cancer prevention (excretion of toxic metabolites).     

A Fluorescent Lectin Test for Mold in Raw Tomato Juice

Fungal (mold) contamination is an important indicator of low quality raw product in the processing tomato industry. A quantitative lectin assay was developed that was less expensive, faster, and more precise than the industry standard Howard mold count. This assay, based on a fluorescent-labeled lectin isolated from wheat germ, had a selective affinity for the chitin in fungal cell walls. Assay values were correlated with mold contamination for 4 fungal species: Alternaria alternata (r²= 0.91), Cladosporium herbarum (r²= 0.75), Fusarium oxysporum (r²= 0.97), and Stemphylium botryosum (r²= 0.99). Combining all 4 species, the lectin assay had a strong correlation (r²= 0.76) with a linearized Howard mold count.     

Botryticidal Activity of Nanosized Silver‐Chitosan Composite and Its Application for the Control of Gray Mold in Strawberry

Botrytis cinerea Pers, the gray mold fungus, is among the most dangerous plant pathogens that cause great losses in agricultural crops. The botryticidal activities of nanosized silver (nano Ag), fungal chitosan (CTS) irradiated fungal chitosan (IrCTS), and nano Ag‐IrCTS composite were investigated. All of the examined materials exhibited powerful antifungal activity against B. cinerea. The most effective compound was nano Ag‐IrCTS composite with a minimal inhibitory concentration of 125 μg/mL. The microscopic examination, of treated B. cinerea with the composite, revealed that an obvious alteration in mycelial shape was appeared accompanied and moderate lysis in fungal hyphae. With the prolongation of treatment, most of the fungal mycelia were lysed into small and elastic fragments. The consequence of strawberries coating, with antifungal composite based solution, was the disappearance of gray mold infection signs in 90% of the contaminated fruits after 7 d of storage, treated fruits had a fresh‐like appearance at the end of storage. Coating with nano Ag‐IrCTS solution could be highly recommended regarding its efficiency in prohibiting B. cinerea growth, preventing gray mold decay and enhancing the overall quality of coated strawberry fruits.     

烟台地区引起甜樱桃采后腐烂霉菌的分离鉴定

为明确引起烟台地区甜樱桃采后腐烂的主要霉菌,对烟台采后腐烂甜樱桃中的霉菌进行分离,应用内部转录间隔区序列分析方法和形态学观察将分离的霉菌进行鉴定。选出代表性霉菌菌株回接樱桃,进行致腐性检测。结果表明:从烟台不同区县收集的腐烂樱桃中分离出的60株霉菌分属于6个种:燕麦赤霉菌(Gibberella avenacea)、总状毛霉菌(Mucor racemosus)、三线镰刀菌(Fusarium tricinctum)、互生链格孢菌(Alternaria alternate)、奥桑青霉菌(Penicillium polonicum)和烟曲霉菌(Aspergillus fumigatus),其中互生链格孢菌和总状毛霉数量较多,分别为14株和15株。将这些霉菌中的代表菌株重新接回到甜樱桃,均能引起甜樱桃的腐烂。     

Effect of chitosan and chitooligosaccharide on vitamin C and polyphenols contents in cherries and strawberries during refrigerated storage

The objective of this research was to investigate the effect of treatment with chitosan and chitooligosaccharide on vitamin C and polyphenols contents in cherries and strawberries during refrigerated storage. Chitosans and chitooligosaccharides, as well as vitamin C and polyphenols are associated with reduced risk of cardiovascular diseases and cancer. Synthesis of vitamin C in strawberries and loss of vitamin C in cherries were observed during refrigerated storage. The treatment with chitosan and chitooligosaccharides inhibits vitamin C synthesis in strawberries and promotes vitamin C synthesis in cherries. Lower values of anthocyanin content for strawberries and higher values of anthocyanin content for cherries treated with chitosan and chitooligosaccharide compared with anthocyanin content in control strawberries and cherries were found after 7 days storage at 4 °C. Total phenols content in strawberries decreased and in cherries increased after storage 7 days at 4 °C and more pronounced changes observed for samples treated with high molecular weight chitosan. The correlation between moisture content in treated and non-treated strawberries and contents of vitamin C, anthocyanin and total phenols was shown. No correlation between moisture content and content of vitamin C, anthocyanins and total phenols was found for cherries.     

Use of ozone to reduce molds in a cheese ripening room

Cheese ripening rooms have an unusual environment, an environment that encourages mold growth. Ozone has been applied in various ways in the food industry. One useful advantage of ozone is that it inactivates molds. In this study, a cheese ripening room was ozonated, and the effectiveness of this treatment was evaluated both in air and on surfaces through sampling on a weekly basis over a 3-month period. The results obtained indicate that ozone treatment reduced the viable airborne mold load but did not affect viable mold on surfaces. Only by wiping the surfaces with a commercial sanitizer was it possible to decrease the viable mold load on surfaces. To improve overall hygiene in the ripening room, a combination of cleaning regimes is recommended. The mold genera occurring most frequently in the air of the cheese ripening room were Penicillium, Cladosporium, and Aspergillus, which accounted for 89.9% of the mold isolates. Penicillium and Aspergillus were identified to the species level, and data showed that P. brevicompactum and P. aurantiogriseum, as well as A. versicolor, were the species most frequently isolated.     

Fluctuations in the Levels of Antioxidant Compounds and Antioxidant Capacity of Ten Small Fruits During One Year of Frozen Storage

The fluctuations in antioxidant capacity (2,2-diphenyl-1-picrylhydrazyl, 2,2´-azinobis (3-ethylbenzthiazoline-6-sulphonic acid), and ferric reducing/antioxidant power assays), total phenol, total flavonoid, and total anthocyanin contents of 10 small fruits (sweet cherries, sour cherries, strawberries, red currants, raspberries, blackberries, hawthorn, cornelian cherries, and red and white grapes) were monitored during storage at ?20°C. After one year in storage, all varieties except hawthorn and white grapes retained equal or slightly greater total phenol content in comparison to the initial values. The total flavonoid and total anthocyanin levels also remained stable or even increased after 12 months in all fruits exept hawthorn and strawberries. Red currants and raspberries exhibited optimal preservation of antioxidant capacity, while hawthorn showed a decrease in the antioxidant capacity during different time points in storage as well as at the end of storage. Analyzed fruits were classified into four main groups based on hierarchical cluster analysis.     

拮抗酵母菌挥发物单体对草莓灰霉病的防治效果

为研究葡萄汁有孢汉逊酵母（Hanseniaspora uvarum）挥发物单体对草莓灰霉病的防治作用,研究了21 种挥发物单体离体条件下对灰葡萄孢的抑制作用,以及活体条件下3-羟基己酸乙酯、癸酸乙酯、桂皮醛和辛酸乙酯对草莓果实灰霉病的防治作用。结果表明：除月桂酸外,其余20 种单体均对灰葡萄孢菌丝生长和孢子萌发有抑制作用,其中桂皮醛剂量为62.5 μL/L时完全抑制菌丝生长,625 μL/L时可以完全抑制孢子萌发;活体条件下,4 种单体均能有效地抑制草莓果实采后灰霉病的发生,保持草莓色泽,延缓草莓可溶性固形物质量分数、可滴定酸质量分数、硬度等质构特性的下降,其中桂皮醛防治草莓灰霉病效果最好,具有商业开发前景。     

猕猴桃广谱拮抗内生菌分离鉴定及其对樱桃圣女果采后的保鲜作用

筛选具有广谱抑菌效果的内生菌并研究其发酵液对樱桃圣女果采后保鲜效果的影响。采用研磨法从健康猕猴桃的根、茎、叶和果实中分离纯化了18株内生菌,通过平板对峙法选出强活性菌株MR-1,并对其进行菌体、菌落形态的观察、生理生化测定及16S rRNA测序分析,进行菌种鉴定。进一步研究该活性菌株对8种常见致病菌的抑制效果以及樱桃圣女果主要病害菌的抑制作用。结果表明,菌株MR-1对采后致病菌拮抗效果较强,初步鉴定其为解淀粉芽孢杆菌,其对8种植物病原菌均有较强的拮抗作用,对链格孢、灰葡萄孢、辣椒炭疽病菌3种圣女果主要病害菌表现出较强的抑制作用,经内生菌MR-1发酵滤液处理的樱桃圣女果保鲜效果最好,其次为复合果蔬保鲜剂。储藏第8 d,与空白对照组相比,其腐烂直径分别下降了50.6%、50.6%及44.9%。解淀粉芽孢杆菌MR-1次级代谢产物丰富,其在果蔬保鲜中有较好的应用前景。