Abstract
Penicillium species belong to main causative agents of food spoilage leading to significant economic losses and potential health risk for consumers. These fungi have been isolated from various food matrices, including table eggs. In this study, both conventional Polymerase Chain Reaction (PCR) and Polymerase Chain Reaction-Internal Transcribed Spacer-Restriction Fragment Length Polymorphism (PCR-ITS-RFLP) methods were used for species identification of Penicillium (P.) spp. isolated from the eggshells of moldy chicken eggs. Seven restriction endonucleases (Bsp1286I, XmaI, HaeIII, HinfI, MseI, SfcI, Hpy188I) were applied to create ribosomal restriction patterns of amplified ITS regions. To identify P. verrucosum, P. commune, and P. crustosum with the help of conventional PCR assay, species-specific primer pairs VERF/VERR, COMF/COMR, and CRUF/CRUR were designed on the base of 5.8 subunit-Internal Transcribed Spacer (5.8S-ITS) region. Altogether, 121 strains of microscopic filamentous fungi were isolated by traditional culture mycological examination. After morphological evaluation of both macroscopic and microscopic features, 96 strains were classified in Penicillium spp. Two molecular methods used have confirmed eight isolates as P. verrucosum, 42 isolates as P. commune, and 19 isolates as P. crustosum. Both PCR-ITS-RFLP and conventional PCR assays appear to be suitable alternatives for rapid identification of the above mentioned Penicillium species.
Highlights
Penicillium spp. is one of the most common microscopic filamentous fungi in foodprocessing industry with more than 200 species known so far [1]
Strains of microscopic filamentous fungi classified as P. verrucosum shown a good growth on MEA, Czapek Yeast Autolysate agar medium (CYA), and Yeast Extract Sucrose agar medium (YES) media, with average colony diameters (25.25 ± 8.84) mm for MEA; (25.42 ± 8.56) mm for CYA and (22 ± 10.41) mm for YES agar after 7 day incubation at 25 ◦C
Penicillium spp. are ubiquitous microscopic filamentous fungi that have been isolated from widespread substrates
Summary
Penicillium spp. is one of the most common microscopic filamentous fungi in foodprocessing industry with more than 200 species known so far [1]. Penicillium species grow best at temperatures between 5 ◦C and 37 ◦C, water activity from 0.78 to 0.88 and pH level from 3 to 4.5 They occur in soil, on decomposing vegetation and compost, on dried food, spices, cereals, in fresh fruits and vegetables, as well as in the air and dust [2]. In particular table eggs, are susceptible to contamination by microscopic filamentous fungi. This contamination can be involved by a variety of sources, e.g., feces, litter, feed, and from improper handling and storage [5]. As temperatures close to +15 ◦C are preferably used in egg storage facilities, the presence of all the three Penicillium species could be predicted in table eggs during storage
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