Beuchat L R
Department of Food Science and Technology, University of Georgia, Griffin 30223-1797.
Int J Food Microbiol. 1992 Oct;17(2):145-58. doi: 10.1016/0168-1605(92)90112-g.
Dilution plating techniques are designed to determine populations of viable fungal propagules per unit weight or volume of food. Direct plating techniques, on the other hand, are designed to assess the internal mycoflora of individual pieces of foods, e.g., seeds or dried fruits, and results are expressed as a percentage of infected pieces. Both techniques are used by industry and regulatory agencies to monitor levels of fungal contamination at various stages of food handling, storing, processing and marketing. Peptone (0.1%) water is commonly used as a diluent for samples to be homogenized or blended. Buffered diluents containing up to 30% glucose or 60% sucrose are recommended for enumerating xerophiles. No one medium is satisfactory for detection or enumeration of yeasts and moulds in all foods. Dichloran rose bengal chloramphenicol agar, oxytetracycline glucose yeast extract agar and rose bengal chloramphenicol agar are superior to acidified potato dextrose agar for enumeration of yeasts and moulds. Dichloran 18% glycerol agar performs well for enumerating moderately xerophilic yeasts and moulds. Fastidious xerophiles require media containing high concentrations of sugars and/or sodium chloride. Media have been formulated to detect potentially aflatoxigenic aspergilli and mycotoxigenic strains of penicillia and fusaria, but increased selectivity and specificity of media for detecting mycotoxigenic moulds are needed. Heat-resistant mould ascospores often require heat treatment prior to plating in order to activate the germination process. The spread-plate technique is strongly preferred over the pour-plate technique for enumerating yeasts and moulds. The recommended incubation temperature is 25 degrees C, but incubation time between plating and counting colonies ranges from 5 days for determination of general populations of mycoflora to 4 weeks or more for fastidious xerophiles. There is a need for new and improved media for selectively isolating various groups, genera, species and/or strains of fungi capable of growing only under specific environmental conditions, e.g., low aw or, in the case of sublethally injured cells, under conditions which facilitate resuscitation. Improved media are needed which accurately detect moulds producing specific mycotoxins in a wide range of food types.
稀释平板技术旨在确定每单位重量或体积食品中 viable 真菌繁殖体的数量。另一方面,直接平板技术旨在评估单个食品(如种子或干果)的内部真菌区系,结果以受感染食品的百分比表示。行业和监管机构都使用这两种技术来监测食品处理、储存、加工和销售各个阶段的真菌污染水平。蛋白胨(0.1%)水通常用作待均质化或混合的样品的稀释剂。建议使用含有高达30%葡萄糖或60%蔗糖的缓冲稀释剂来计数嗜干菌。没有一种培养基能令人满意地检测或计数所有食品中的酵母和霉菌。二氯苯胺玫瑰红氯霉素琼脂、土霉素葡萄糖酵母提取物琼脂和玫瑰红氯霉素琼脂在计数酵母和霉菌方面优于酸化马铃薯葡萄糖琼脂。二氯苯胺18%甘油琼脂在计数中度嗜干酵母和霉菌方面表现良好。苛求嗜干菌需要含有高浓度糖和/或氯化钠的培养基。已经配制了培养基来检测潜在产黄曲霉毒素的曲霉菌以及青霉菌和镰刀菌的产霉菌毒素菌株,但需要提高检测产霉菌毒素霉菌的培养基的选择性和特异性。耐热霉菌子囊孢子在平板接种前通常需要进行热处理以激活萌发过程。在计数酵母和霉菌时,强烈推荐使用涂布平板技术而不是倾注平板技术。建议的培养温度为25℃,但从接种到计数菌落的培养时间范围从用于确定真菌区系总体数量的5天到用于苛求嗜干菌的4周或更长时间。需要新的和改进的培养基来选择性分离仅在特定环境条件下(如低水分活度)或对于亚致死损伤细胞在有利于复苏的条件下能够生长的各种真菌群、属、种和/或菌株。需要改进的培养基来准确检测在广泛食品类型中产生特定霉菌毒素的霉菌。