Identification Of Fungi On The Surface Of Five Health-Care Chinese Herbal Materials

Traditional Chinese medicine is the treasure of the Chinese nation.Health-care Chinese herbal materials occupy a key position in the food industry and modern medical system in China.Recently.the contaminations of fungi and mycotoxins in Chinese herbal materials have been widely concerned,posing a potential threat to consumer health and the industry development.This study identified the fungi on the surface of five Healthcare Chinese herbal materials through DNA metabarcoding technology.The total DNA on the surface of the materials was exacted.The polymerase chain reaction(PCR)of nuclear ribosomal RNA internal transcribed spacer 2(ITS2)was amplified by using the primer pairs.The ITS2 region was sequenced through the Illumina Miseq PE300 platform.After the sequences were purified and assembled,cluster analysis was conducted.Fungal diversity,fungal composition,and comparison of fungal community were analyzed.The following was the contents and conclusions:1.Identification of fungi on the surface of Angelicae Sinensis Radix(ASR)was conducted.The 15 ASR samples were collected from five main production areas.Results showed that 15 samples were contaminated by fungi.Ascomycota,Leotiomycetes,Helotiales,and Didymellaceae were the dominant fungi,the relative abundances were 14.89%-99.57%,0.19%-84.97%,0.19%-65.60%.and 0.26%-74.68%respectively.Cladosporium,Fusarium,and Penicillium were distributed in all samples.Besides ASRY3 and ASRH1,Aspergillus and Rhizopus were detected in all the other samples.Furthermore,there were four toxigenic fungi(Penicillium oxalicum P.brevicompactum,P.citrinum,and Aspergillus clavatus)and one human pathogen(Sarocladium strictum).2.Identification of fungi on the surface of Panacis Quinquefolii Radix(PQ)was conducted.The fungi were detected in 15 samples that were collected from five main production areas.Mucoromycota(13.53%-57.81%).Mucoromycetes(13.86%-57.81%),Mucorales(13.53%-57.81%),Rhizopodaceae(13.86%-57.79%)were predominant.Rhizopus(13.86%-57.79%)was the most abundant,followed by Aspergillus(6.05%35.59%)and Fusarium(2.62%-12.44%).Except for PQ_BJ1 and PQ_SX3.Penicillium was distributed in all PQ samples,and the relative abundance in PQ_SD2 was the highest.The results indicated six human pathogens:Candida tropicalis.C.parapsilosis,C.palmioleophila,C.zeylanoides,Malassezia restricta,and M.globose.3.Identification of fungi on the surface of Polygoni Multiflori Radix(PMR)was conducted.There were 18 samples collected from five different areas in China,including 15 PMR samples and three Polygoni Multiflori Radix Praeparata(PMRP)samples.The groups based on the collection areas and processing methods were different significantly.18 samples were contaminated by fungi.Ascomycota,Eurotiomycetes,Eurotiales,Aspergillaceae and Aspergillus were dominant at phylum,order,and genus levels,with the relative abundance of 71.55%-99.77%,6.30%-98.07%,2.57%-98.07%.3.77%97.91%,and 2.14%-94.05%.The result showed six toxigenic fungi(Aspergillus flavus,A.nomius,A.tamarii,Fusarium solani,Penicillium paxillin,and P.steckii)and one human pathogen(Candida tropicalis).4.Identification of fungi on the surface of Fritillariae Cirrhosae Bulbus(FCB)was conducted.The 15 FCB samples,collected from five provinces,were detected the existence of fungi.Ascomycota(41.58%-99.66%),Eurotiomycetes(8.49%-63.93%).Eurotiales(8.49%-63.53%).and Aspergillaceae(8.49%-63.51%)were dominant at the phylum,class,order,and family level.Aspergillus(8.49%-63.41%).Rhizopus(0%57.42%),and Fusarium(0%-22.81%)were the predominant genus.There were five toxigenic fungi detected(Penicillizum brevicompactum,P.citrinum.P.oxalicum.Trichothecium roseum,and Aspergillus rest rictus).5.Identification of fungi on the surface of Lonicerae Japonicae Flos(LJF)was conducted.A total of 18 LJF samples,collected from five main production areas,were detected fungal contaminations.The groups based on the collection areas and processing methods were different significantly.Ascomycota(20.77%-88.41%),Mucoromycetes(6.40%-79.23%),Mucorales(6.40%-79.23%),and Rhizopodaceae(6.40%-79.23%)were predominant.Rhizopus and Aspergillus contaminated all samples.Besides LJFHB3,Fusarium was detected in 17 samples.Moreover.ten human pathogens were detected:Candida tropical is,C.parapsilosis,C.sake,Malassezia restricta,M.sympodialis.Kodamaea ohmeri,Lodderomyces elongisporus,Schizophyllum commune,Wallemia sebi,and Mucor circinelloides.This study could provide references for supervising the fungal contamination in health care medicine,and offer an early warning for the better control mycotoxin contaminations.The research had a significant meaning in guaranteeing the standardized production of Chinese herbal materials and ensuring public safety drug use.