Identification and Molecular Characterization of Bacteria and Fungi Associated with Three Fresh Edible Mushrooms

Anyakorah Caroline Iruoma, Adeniji Omolara Folakemi
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Abstract

Aims: Mushrooms are health-food used as food supplements and fortification but are susceptible to microbial spoilage due to high moisture content and nutrient status. Fresh mushrooms sometimes show signs of spoilage after harvest and makes storage difficult. The objective of the study was to identify and characterize bacteria and fungi associated with three fresh edible mushrooms, under ambient and cold temperatures. Methodology: The study was conducted at the Bells University of Technology, Nigeria, between December 2020 and October 2021. Pleurotus ostreatus and Calocybe indica fruitbodies were procured from the Federal Institute of Industrial Research Oshodi while Pleurotus tuber-regium fruitbodies were obtained from sclerotia planted in loamy soil. The fruitbodies were kept at ambient (28oC) and cold (15oC) temperatures respectively. The bacterial and fungal counts on each of the mushrooms were taken at 0,3, 5, and 7 days after harvest. The isolated bacteria were identified by conventional methods; Analytical Profile Index (API) 20E kits (BioMerieux), while Fungi were identified by morphological features and PCR amplification using ITS 1f/ ITS 4r universal primers.  Results: The bacterial and fungal counts on the fruitbodies ranged from 5.7 log CFU/ml – 6.3 log CFU/ml and 5.0 log CFU/ml – 5.9 log CFU/ml respectively. Seven genera of bacteria isolated were gram-negative bacteria. At ambient temperature, Pseudomonas aeruginosa, Enterobacter asburiae, Klebsiella oxytoca, Klebsiella ornithinolytica, Serratia marcescens, Chryseobacterium meningosepticum and Cedecea davisae were isolated while Enterobacter cloacae, Enterobacter sakazakii and Citrobacter braakii were isolated at cold temperature. Aspergillus, Penicillium, and Fusarium were isolated at both temperatures while Alternaria was isolated at ambient temperature. Conclusion: Isolated bacteria and fungi were mostly enteric pathogens and potential mycotoxin-producing fungi. This is an indication that strict hygiene and control measures should be put in place during the production and storage of these mushrooms in order to improve the quality and food safety of fresh mushrooms in Nigeria.
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三种新鲜食用菌相关细菌和真菌的鉴定及分子特性研究
目的:蘑菇是用作食品补充和强化的保健食品,但由于其高水分含量和营养状况,容易受到微生物的破坏。新鲜的蘑菇有时会在收获后出现变质的迹象,使储存变得困难。该研究的目的是在常温和低温下鉴定和表征与三种新鲜食用蘑菇有关的细菌和真菌。方法:该研究于2020年12月至2021年10月在尼日利亚贝尔科技大学进行。平菇和Calocybe indica子实体来自Oshodi联邦工业研究所,而块茎-区域平菇子实体来自壤土种植的菌核。子实体分别在常温(28℃)和低温(15℃)下保存。在收获后0、3、5和7天对每种蘑菇进行细菌和真菌计数。采用常规方法对分离细菌进行鉴定;真菌采用ITS 1f/ ITS 4r通用引物,通过形态学特征和PCR扩增进行鉴定。结果:子实体细菌和真菌计数范围分别为5.7 ~ 6.3 log CFU/ml和5.0 ~ 5.9 log CFU/ml。分离出革兰氏阴性菌7属。常温下分离出铜绿假单胞菌、土产肠杆菌、氧化克雷伯菌、禽毒克雷伯菌、粘质沙雷菌、脑膜炎败血症黄杆菌和鸟粪菌,低温下分离出阴沟肠杆菌、坂崎肠杆菌和布拉克柠檬酸杆菌。在两种温度下分离曲霉、青霉和镰刀菌,而在室温下分离交替菌。结论:分离的细菌和真菌以肠道致病菌和潜在的产霉菌毒素真菌为主。这表明,在这些蘑菇的生产和储存过程中,应该采取严格的卫生和控制措施,以提高尼日利亚新鲜蘑菇的质量和食品安全。
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