Identification and biodiversity patterns ofAspergillusspecies isolated from some soil invertebrates at high altitude using morphological characteristics and phylogenetic analyses

Abstract

BackgroundThe carcinogenic, mutagenic, and teratogenic chemicals such as aflatoxin are a worldwide health problem.Aspergillusspp., responsible for most cases of aflatoxin contamination, are common in the environment and spread easily to many different types of food. The objectives of this study were to conduct a survey of fungi associated with three soil invertebrates in Taif, Saudi Arabia, identify these isolates and explore mycotoxins formation.MethodsIn total, 114 fungal isolates were collected from various soil invertebrates (millipedes,Armadillidium vulgareandPorcellio laevis) in Taif, Saudi Arabia, among them, 22 isolates were identified asAspergillusspp. based on morphological and molecular characteristics followed by bothFusariumandPenicillium.ResultsThe sequences of ITS 1 and ITS 4 were utilized. Using bootstrap analysis, phylogenetic tree was split into two distinct clusters. Five sub clusters were included inside the first major cluster, and their bootstrap value was 99%. While, there were two small clusters in the second major cluster. All the testedAspergillusstrains were able to have a single PCR fragment amplified using the primer AspTef. TEF-1 DNA sequence bootstrap analysis with 1,000 replicates revealed two distinct groups. Additionally, theAspergillusisolates were grouped into two different clusters with about 65% genetic similarity using ISSR-PCR analysis. The standard polymerase chain reaction was used to effectively amplify theAopks, afl-Aandomt-A genes in aflatoxigenicAspergillusstrains. FourAspergillusstrains used in this investigation were shown to generate aflatoxin B1. While, threeAspergillusstains showed ochratoxin genes.ConclusionsIn conclusion, the results indicate significant differences in the fungal community between ecoregions and soil invertebrates. Moreover, mycotoxin detection and identification amongAspergillusisolates were elucidated. This study could shed light on the risk of mycotoxin contamination along the supply chain.