PSYCHROTROPHIC AND MESOPHILIC FUNGI ISOLATED FROM IMPORTED FROZEN LEAN MEAT IN EGYPT

From 475 samples of imported frozen lean beef in Egypt, 859 psychrotrophic fungal strains (770 strains by the dilution plating and 89 strains by direct plating) were isolated. There were also 1291 mesophilic fungal strains isolated (1133 and 158 strains by the two different methods, respectively). The predominant psychrotrophs identified were species of Cladosporium and Penicillium while most mesophiles were in the genera of Aspergillus, Cladosporium and Penicillium.     

FUNGAL BIOTA ISOLATED FROM SPANISH CHEESES

Fungal biota, with special reference to the genus Penicillium, was studied in 52 samples of commercial cheeses (10 fresh, 17 semiripened and 25 ripened) made from different types of milk (cow, ewe, goat and mixed) produced in southern Spain. In 41 of the total of cheeses analyzed (79%) molds were isolated. Penicillium was identified in 63% of the samples , Mucor spp. in 27% , Geotrichum candidum in 17% and Cladosporium herbarum in 10%; eleven other fungal genera were detected ranging from 2 to 4%. Thirty-five species of Penicillium were analyzed with the following distribution: 7 in fresh cheese, 16 in semiripened cheese and 30 in ripened cheese. The incidence of Penicillium spp. was also greater in the cheeses with a higher degree of ripeness, i.e. 20% in fresh cheese, 71% in the semiripened and 76% in the ripened cheese.     

Moulds and yeasts in fresh and minimally processed vegetables, and sprouts

A limited survey of fresh and minimally processed vegetables, and sprouts was conducted in the Washington, DC area to determine if potentially toxigenic and pathogenic fungi were present in these commodities. Thirty-nine ready-to-eat salads, 29 whole fresh vegetables and 116 sprout samples (bean, alfalfa, broccoli, crunchy, garlic, spicy, onion, clover, lentil and multi-seed sprouts) were purchased from 13 local supermarkets and tested for yeast and mould counts as well as the presence of toxigenic moulds. Yeasts were the most prevalent organisms found in these samples, at levels ranging from less than 100 to 4.0x10(8) cfu/g. Mould counts generally ranged from less than 100 to 4.0x10(4) cfu/g. Two crunchy sprout samples, however, contained unusually high numbers of Penicillium (1.1x10(8) and 1.3x10(8) cfu/g), two alfalfa sprout samples contained Geotrichum populations about 10(6) cfu/g, and two alfalfa sprout samples had Cladosporium counts higher than 2.5x10(5) cfu/g. The most common moulds found in fresh and minimally processed vegetables were Cladosporium, Alternaria and Penicillium; less common was Geotrichum. The most frequently isolated moulds from sprouts were Alternaria, Cladosporium, Penicillium, and Phoma. Phoma was especially common in alfalfa sprouts. Fusarium, Rhizopus, Mucor, and Geotrichum were isolated less often.     

Use of the selective agar medium CREAD for monitoring the level of airborne spoilage moulds in cheese production

It was investigated if a selective medium for common cheese spoiling moulds (CREAD) could give more relevant information than a general mould medium in hygienic air-sampling in cheese factories. A total of 126 air-samples were taken in six Nordic cheese factories using the general mould medium DG18 and CREAD. The level and genera of air-borne mould was determined. Identification to species-level was performed for a selection of samples. In five cheese factories the mycobiota was dominated by Penicillium spp. and in one cheese factory by Cladosporium spp. The concentration of air-borne moulds varied between the cheese factories ranging from 1 to 270 cfu/m3 on DG18 with a median value of 17. The number of mould colonies was in general lower at CREAD. Identification indicated that CREAD supported growth of common spoilage moulds for cheese, such as Penicillium palitans and P. commune. The mycobiota on DG18 also consisted of moulds not commonly associated with spoilage of cheese, such as Cladosporium spp., P. brevicompactum and P. chrysogenum. Contamination of cheese with mould is periodically a problem in production of semi-hard cheese and the level of air-borne mould is therefore routinely monitored in cheese factories. A clear correlation between the total number of moulds in air and mould growth on products is not always found. The conclusion from the investigation is that it is recommended to use a selective medium for cheese spoilage moulds, such as CREAD in hygienic monitoring.     

Effect of processing for saponin removal on fungal contamination of quinoa seeds (Chenopodium quinoa Willd.)

Incidence of fungal contamination of quinoa seeds from three locations (Salar de Uyuni, Bolivia; Salta and Tucumán provinces, Argentina) was analyzed in samples with and without treatment to remove saponins (wet method). In processed samples, the percentage of infection was reduced. Distribution of the different fungal genera was not homogeneous in the three locations (p<0.05), although Penicillium and Aspergillus were the most prevalent contaminants, regardless the geographic origin of the samples. Other genera, such as Eurotium, Fusarium, Phoma, Ulocladium, Mucor and Rhizopus were less frequently isolated. Absidia, Alternaria, Cladosporium, Dreschlera, Epicoccum and Monascus were sporadically encountered. Significant differences (p<0.05) in the distribution of fungal genera in samples with and without saponins from each location were observed. In all cases, processing caused a decrease of Aspergillus incidence, while increased the proportion of Penicillium, Eurotium, Mucor and Rhizopus indicating that these genera were part of the internal mycota. A. flavus and A. niger were the dominating species of genus Aspergillus. A similar pattern of prevalent Penicillium species was observed in samples with and without saponins, since P. aurantiogriseum, P.chrysogenum, P. citrinum and P. crustosum were always present in high number, although their relative density was variable according to the geographic origin of samples. Mycotoxin-producing ability of most representative species was also determined. Toxigenic strains of A. flavus (aflatoxins and cyclopiazonic acid), A. parasiticus (aflatoxins), P. citrinum (citrinin) and P. griseofulvum (cyclopiazonic acid) were found. None of the A. niger isolates was ochratoxin A producer. The above mentioned mycotoxins were not detected in the samples analyzed.     

A SURVEY OF OCHRATOXIN A IN WHEAT AND BARLEY IN INDIA

Ochratoxin A (OTA), a secondary metabolite produced by the species of Aspergillus and Penicillium has been found to be nephrotoxic, immunosuppressive and teratogenic in many species and is a possible human carcinogen. Its prevalence in cereals, namely wheat and barley, has been reported from many European countries. In this study, 126 samples of wheat and barley collected from different regions in India were analyzed by high‐performance liquid chromatography for the presence of OTA. The highest OTA content determined in one sample of wheat was 2.3 µg/kg. None of the 126 samples of wheat and barley contained more than 5 µg/kg. Wheat (82.6%) and barley (90%) samples contained less than 0.1 µg/kg, which is the limit of quantification of OTA in this study. The maximum limit of OTA for wheat, barley and rye that is being debated at Codex Alimentarius Commission meetings is 5 µg/kg. In an attempt to determine if there was any correlation between the ochratoxigenic mold mycoflora and the ochratoxin content in the samples analyzed, enumeration of the molds that belonged to Aspergillus and Penicillium species was determined using general mycological media. The aflatoxigenic and ochratoxigenic mycoflora was also enumerated using selective agar media. It was observed that more aflatoxigenic molds were recovered in both wheat and barley samples. None of the species of Penicillium detected from the samples was found to be ochratoxigenic. However, Aspergillus species belonging to section Nigri, Aspergillus niger group, also an ochratoxin producer, were recovered from the samples.     

一例茶饮料污染源的分析

某企业生产的一批茶饮料中发现某些产品变质,3份样品出现pH值下降,或产气胀罐现象,或有肉眼可观察到浑浊、变色等感官指标的变化。检测发现引起腐败变质的主要是细菌,其数量≥104 CFU/mL,4株主要污染菌菌株经16S rDNA序列比对,鉴定为假单胞菌属(Pseudomonas sp.)、肠杆菌属(Enterobacter sp.)、芽胞杆菌属(Bacillus sp.)和短芽孢杆菌(Brevibacillus sp.)。样品中还有数量很低(101～102 CFU/mL左右)的霉菌检出,经形态观察初步鉴定为交链孢霉(Alternaria spp.)、枝孢霉(Cladosporium spp.)、曲霉(Aspergillus spp.)和青霉(Penicillium spp.)。该案例中霉菌虽然不是主要的腐败变质菌,但是其菌相组成与空气中主要的霉菌菌相一致,显示该污染的源头可能是空气,细菌的组成也能支持这个推论。实践中,通过更换无菌空气过滤器,确实解决了污染问题,证实了该推论。     

紫薯腐败菌的分离与鉴定

目的 从自然腐败的紫薯上分离纯化紫薯腐败菌, 通过反接种确定其致腐性, 并将其鉴定到属。方法 通过多次稀释平板涂布培养, 进行单菌落形态学观察, 将菌种反接种到新鲜紫薯上, 筛选出腐败菌, 以真菌ITS序列、细菌16S rDNA序列构建发育树, 鉴定到属, 再将鉴定结果与分离的菌种的形态学鉴定结果对比, 验证菌种鉴定结果。结果 从样品中分离出6种典型腐败菌, 有4种属, 分别为枝孢霉属、青霉属、曲霉属、芽孢杆菌属。结论 确定紫薯的腐败菌为哥氏枝孢霉(Cladosporium gossypiicola)、Penicillium christenseniae、热带青霉(Penicillium tropicum)、黄曲霉(Aspergillus flavus)、贝莱斯芽孢杆菌(Bacillus velezensis)和阿氏芽孢杆菌(Bacillus aryabhattai)。     

CONTAMINATION OF GRAINS BY MYCOTOXIN-PRODUCING MOLDS AND MYCOTOXINS AND CONTROL BY GAMMA IRRADIATION

Ninety random grain samples were collected and analyzed for mycotoxins, and the effect of gamma irradiation on the production of mycotoxins in grains was studied. Aspergillus, Penicillium, Mucor, Rhizopus, Fusarium, Alternaria, Scopulariopsis and Cladosporium were the most common fungal genera isolated from grains. Aspergillus flavus, Aspergillus niger, Aspergillus candidus, Aspergillus ochraceus, Penicillium citrinum, Penicillium expansum, Penicillium citreonigrum, Penicillium purpurogenum, Penicillium griseofulvum and Penicillium verrucosumwere the most common Aspergillus and Penicillium species in grains. Out of 120 Aspergillus and Penicillium isolates, 80 were mycotoxin producers. Analysis of grains revealed the occurrence of aflatoxin B₁ ochratoxin A, cycolopiazonic acid and citrinin. Of the 90 samples, 67 were positive for one or more mycotoxin. Irradiation of grains at dose of 2.0 and 4.0 kGy decreased significantly the total fungal counts compared with unirradiated controls. After 100 days of storage at room temperature, the unirradiated grains were contaminated with high concentrations of mycotoxins as compared with irradiated 4.0‐kGy samples. Mycotoxin production in grains decreased with increasing irradiation doses and was not detected at 6.0 kGy over 100 days of storage.     

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.     

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.     

Microbial quality and presence of moulds in Kuflu cheese

The chemical and microbial qualities, including fungal flora, of 30 samples of Kuflu cheese randomly purchased from different markets in Turkey were investigated. The gross composition of the cheese samples ranged between 37.65-53.65% moisture, 6.21-40.09% fat-in-dry matter, 4.70-10.07% salt-in-moisture and 26.18-44.85% protein. The mean pH value of the cheeses was 6.29+/-0.28 and pH values ranged from 5.52 to 7.22. Variations between the samples in terms of their gross composition suggested a lack of quality standards in cheesemilk, cheesemaking procedure and ripening conditions. The levels of main microbial groups including total mesophilic and coliform bacteria, yeasts and moulds and the presence of some potentially pathogenic microorganisms (E. coli, Salmonella spp. and Staphylococcus aureus) were determined. The high numbers of all microbial groups and presence of potentially pathogenic organisms in the cheese samples suggested that the production and maturation of Kuflu cheese should be improved by better hygiene. Moulds at the cheese surface were isolated and identified. A total of 24 different mould species were detected and the genus most frequently isolated was Penicillium spp. which represented 70.25% of total isolates. Penicillium commune, P. roqueforti and P. verrucosum were the most abundant species in the cheeses sampled. The other dominant fungal groups were Geotrichum candidum, Penicillium expansum and P. chrysogenum. Other genera isolated from the cheese were Acremonium, Alternaria, Aspergillus, Cladosporium, Geotrichum, Mucor, Rhizopus and Trichoderma. The potentially toxigenic species, including some Penicillum spp. and Aspergillus flavus, were also detected.     

烟草种子携带病原真菌的分离与鉴定

采用无菌滤纸培养烟苗的方法对贵州主栽的4个烤烟品种裸种进行了带菌检测,并对种子携带真菌进行了分离与鉴定。结果表明,K326、贵烟4号、韭菜坪2号和南江3号的种子带菌率分别为15%、77.78%、21.67%和61.39%。分离和鉴定发现种子携带的真菌有链格孢菌属（Alternaria sp.）、拟茎点霉属（Phomopsis sp.）、支顶孢属（Acremonium sp.）、柄孢壳属（Zopfiella sp.）、曲霉属（Aspergillus sp.）、镰孢菌属（Fusarium sp.）、青霉菌属（Penicillium sp.）、赤霉菌属（Gibberella sp.）、芽枝孢菌属（Cladosporium sp.）、离蠕孢属（Bipolaris sp.）和Stagonosporopsis cucurbitacearum等。获得40株真菌中共有22种真菌,优势菌群主要为链格孢属、芽枝孢菌属、镰孢菌属和赤霉菌属。链格孢菌属在4个烤烟品种上均有发现。镰孢菌属主要来自于贵烟4号、韭菜坪2号和南江3号,其中南江3号种子上最多。芽枝孢菌属仅来自贵烟4号。贵烟4号和南江3号种子上的真菌种类和数量较多。这些种子携带真菌的发现对烤烟种子的生产与加工具有重要的意义。     

Fungi causing thread mould spoilage of vacuum packaged Cheddar cheese during maturation.

Thread mould is a defect which occurs sporadically in maturing vacuum packaged Cheddar cheese, caused by the growth of fungi in folds and wrinkles of the plastic film in which the cheese is packaged. Fungi were isolated and identified from 110 Cheddar cheese blocks exhibiting typical thread mould defects. The major causative species were found to be Cladosporium cladosporioides, Penicillium commune, C. herbarum, P. glabrum and a Phoma species. Yeasts were also frequently isolated from the cheese, the majority belonging to the genus Candida. Fungal species which can cause thread mould defects were also found in the cheese factory environment, on cheesemaking equipment, in air, and in curd and whey, providing a wide range of potential sources of contamination.     

The presence of Penicillium and Penicillium mycotoxins in food wastes

A total of 97 samples (48 summer and 49 winter) of food waste from private households were investigated for Penicillium and for mycotoxins. Twenty-five Penicillium species were isolated and Penicillium crustosum, Penicillium brevicompactum, Penicillium chrysogenum, Penicillium expansum, Penicillium roqueforti, Penicillium spinulosum, Penicillium viridicatum, Penicillium commune, Penicillium citrinum and Penicillium solitum were, in decreasing order, the most frequently identified species. Mycotoxins produced by several of these species, including mycophenolic acid, roquefortine C, penitrems A-F and thomitrems A and E, were detected. Of the 48 summer samples, 36 were severely infected and contained more than 10(5) colony forming units (CFU) Penicillium/g sample. The levels of mycotoxins in these samples were in the range 75-19000 microg/kg mycophenolic acid, 40-920 microg/kg roquefortine C, 35-7500 microg/kg penitrem A, 20-2100 microg/kg thomitrem A and 20-3300 microg/kg thomitrem E. Of the 49 winter samples, only one was found to contain mycophenolic acid (4800 microg/kg) and roquefortine C (190 microg/kg), and this sample was severely infected with P. roqueforti. Thirty samples of food waste collected from the food manufacturing industry were also investigated. The number of Penicillium in these samples was between 10(5) and 10(6) colony forming units (CFU)/g sample. Seven of these samples contained mycophenolic acid ranging from 50 to 600 microg/kg and three of these samples also contained roquefortine C in the range 100-250 microg/kg.     

Moulds and yeasts in fruit salads and fruit juices

Thirty-eight fruit salad samples including cantaloupe, citrus fruits, honeydew, pineapple, cut strawberries and mixed fruit salads, and 65 pasteurized fruit juice samples (apple, carrot, grapefruit, grape and orange juices, apple cider, and soy milk) were purchased from local supermarkets in the Washington, DC area and tested for fungal contamination. The majority of fruit salad samples (97%) were contaminated with yeasts at levels ranging from <2.0 to 9.72 log10 of colony forming units per gram (cfu/g). Frequently encountered yeasts were Pichia spp., Candida pulcherrima, C. lambica, C. sake, Rhodotorula spp., and Debaryomyces polymorphus. Low numbers of Penicillium spp. were found in pineapple salads, whereas Cladosporium spp. were present in mixed fruit and cut strawberry salads. Twenty-two per cent of the fruit juice samples tested showed fungal contamination. Yeasts were the predominant contaminants ranging from <1.0 to 6.83 log10 cfu/ml. Yeasts commonly found in fruit juices were C. lambica, C. sake, and Rhodotorula rubra. Geotrichum spp. and low numbers of Penicillium and Fusarium spp. (1.70 and 1.60 log10 cfu/ml, respectively) were present in grapefruit juice.     

不同储藏年限稻谷真菌群落多样性与优势菌属探究

为有效防控储藏稻谷霉变,确保稻谷在储藏期的品质与安全,本研究先后对同一粮库点新入库、储藏1年、储藏2年及储藏3年稻谷样品进行了采集,并对其真菌群落多样性进行了探究。通过Illumina MiSeq序列分析法,探究了不同储藏年限稻谷的真菌群落多样性,优势菌属以及真菌群落随储藏年限增加的演变规律。结果发现新入库储藏稻谷的真菌群落多样性远大于储藏期稻谷。此外,储藏稻谷的优势菌属包括假丝酵母菌属（Gibberella）,曲霉属（Aspergillus）,帚枝霉属（Sarocladium）,枝孢霉属（Cladosporium）,链格孢属（Alternaria）和青霉属（Penicillium）等。储藏年限不同,对应的优势菌属不同,其中曲霉属均是储藏期稻谷的优势菌属之一。     

Mycological survey for potential aflatoxin and ochratoxin producers and their toxicological properties in harvested Brazilian black pepper.

A mycological survey was carried out on 115 samples of whole dried black pepper seeds, from two main production regions of Brazil (Pará and Espírito Santo). A high incidence of contamination was verified in both regions when 99.1% of the samples showed filamentous fungi contamination. A total of 497 species of nine different genera were isolated (Aspergillus, Eurotium, Rhizopus, Penicillium, Curvularia, Cladosporium, Absidia, Emericella and Paecilomyces). The genus Aspergillus was the predominant (53.5%) followed by species from the Eurotium genus (24.5%). Eurotium chevalieri (16.4%) was the most predominant species followed by A. flavus (14.6%) present on 55 samples of black pepper (47.8%) analysed. Twenty-five samples (21.7%) were contaminated with aflatoxigenic strains of A. flavus and A. parasiticus. In relation to the types of aflatoxins produced by mycotoxigenic strains, it was observed that 25 strains (44.6%) of 56 isolated of A. flavus produced aflatoxins. From 12 samples, A. ochraceus species were isolated in low frequency (3.5%). Two strains of A. ochraceus from 16 isolated were producers of ochratoxin A. With respect to the aflatoxins and ochratoxin A natural contamination, none of the samples presented detectable levels of these mycotoxins using thin-layer chromatographic analysis.     

Fungi in desiccated coconut

Twenty-eight fungal species were isolated from desiccated coconut and identified. Members of the genus Eurotium (Aspergillus glaucus group) were the principle species, with Penicillium, Cladosporium, Aspergillus flavus and Aspergillus niger commonly present. Two rare moulds, Chrysosporium farinicola (Burnside) Skou C. fastidium Pitt) and a new species of Arthropsis, were found. Different species were isolated with different diluents. The need for two to three weeks incubation period is discussed and the implications of this for the microbiological specifications for dried foods noted.     

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。