Laut Seavchou, Poapolathep Saranya, Piasai Onuma, Sommai Sujinda, Boonyuen Nattawut, Giorgi Mario, Zhang Zhaowei, Fink-Gremmels Johanna, Poapolathep Amnart
Department of Pharmacology, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand.
Department of Plant Pathology, Faculty of Agriculture, Kasetsart University, Bangkok 10900, Thailand.
Foods. 2023 Jan 20;12(3):487. doi: 10.3390/foods12030487.
The study focused on the examination of the different fungal species isolated from commercial rice samples, applying conventional culture techniques, as well as different molecular and phylogenic analyses to confirm phenotypic identification. Additionally, the mycotoxin production and contamination were analyzed using validated liquid chromatography-tandem mass spectrometry (LC-MS/MS). In total, 40 rice samples were obtained covering rice berry, red jasmine rice, brown rice, germinated brown rice, and white rice. The blotting paper technique applied on the 5 different types of rice samples detected 4285 seed-borne fungal infections (26.8%) for 16,000 rice grains. Gross morphological data revealed that 19 fungal isolates belonged to the genera (18 of 90 isolates; 20%) and (72 of 90 isolates; 80%). To check their morphologies, molecular data (fungal sequence-based BLAST results and a phylogenetic tree of the combined ITS, , , and datasets) confirmed the initial classification. The phylogenic analysis revealed that eight isolates belonged to and, additionally, one isolate each belonged to , , and Furthermore, four isolates of and one isolate of each taxon were identified as (, , and ). The results showed that and were two commonly occurring fungal species in rice samples. After subculturing, ochratoxin A (OTA), generated by code W3-04, was discovered using LC-MS/MS. In addition, the toxin beauvericin was detected in one of the samples. Aflatoxin B1 or other mycotoxins, such as citrinin, trichothecenes, and fumonisins, were detected. These preliminary findings should provide valuable guidance for hazard analysis critical control point concepts used by commercial food suppliers, including the analysis of multiple mycotoxins. Based on the current findings, mycotoxin analyses should focus on toxins, including OTA and metabolites of (recently considered a producer of emerging mycotoxins) to exclude health hazards related to the traditionally high consumption of rice by Thai people.
该研究聚焦于对从市售大米样本中分离出的不同真菌种类进行检测,运用传统培养技术以及不同的分子和系统发育分析方法来确认表型鉴定。此外,使用经过验证的液相色谱 - 串联质谱法(LC-MS/MS)分析霉菌毒素的产生和污染情况。总共获取了40个大米样本,涵盖了稻粒、红米茉莉香米、糙米、发芽糙米和白米。应用于5种不同类型大米样本的吸水纸技术在16000粒大米中检测到4285例种子携带真菌感染(26.8%)。总体形态学数据显示,90株真菌分离物中有19株属于曲霉属(18株;20%)和青霉属(72株;80%)。为检查它们的形态,分子数据(基于真菌序列的BLAST结果以及ITS、cal、tef和tub数据集组合的系统发育树)证实了初步分类。系统发育分析表明,8株分离物属于曲霉属,此外,各有1株分离物属于青霉属、镰刀菌属和枝顶孢属。此外,4株曲霉属分离物和每个分类单元各1株分离物被鉴定为产毒菌株(黄曲霉、赭曲霉和寄生曲霉)。结果表明,曲霉属和青霉属是大米样本中两种常见的真菌种类。继代培养后,使用LC-MS/MS发现由编号W3-04的赭曲霉产生了赭曲霉毒素A(OTA)。此外,在其中一个样本中检测到了青霉毒素博韦里霉素。未检测到黄曲霉毒素B1或其他霉菌毒素,如桔霉素、单端孢霉烯族毒素和伏马菌素。这些初步研究结果应为商业食品供应商使用的危害分析关键控制点概念提供有价值的指导,包括多种霉菌毒素的分析。基于当前研究结果,霉菌毒素分析应侧重于赭曲霉毒素,包括OTA以及赭曲霉的代谢产物(最近被认为是新兴霉菌毒素的生产者),以排除与泰国人传统上大量食用大米相关的健康危害。