Mycotoxigenic fungi contamination of grains and peanuts from open markets in Kelantan, Malaysia

The warm weather and high relative humidity in Malaysia are ideal for the survival and proliferation of mycotoxigenic fungi leading to a high rate of stored product contamination. This study was conducted to enumerate and characterise the mycotoxigenic fungi associated with commonly consumed food grains in Kelantan, Malaysia. The fungal bioburden and fungal identification from forty-four composite food samples comprising 11 samples each of maize, wheat, rice, and peanuts from open markets in Kelantan, Malaysia, were determined using standard mycological techniques. A total of 115 mould fungal isolates belonging to 12 species were isolated, of which Aspergillus flavus (17.39%), A. versicolor (13.04%), A. felis (12.17%), Neoscytalidium dimidiatum (11.3%), Penicillium cheresanum (11.3%) and P. chrysogenum (8.7%), were predominant. Peanuts were the most contaminated (9.7×105 ± 1.5×105 CFU/g) followed by maize (7.5×105 ± 1.8×106 CFU/g), wheat (1.9×105 ± 2.6×105 CFU/g), and rice (9.9×104 ± 1.5×105 CFU/g). The levels of the mycotoxigenic fungi in peanut, maize, and wheat were above the permissible limit of 102 CFU/g set by the Malaysian Ministry of Health and 102 to 105 CFU/g set by the International Commission for Microbiological Specification for Foods, signifying that they are unsafe for use as food or feed ingredients. Hence, there is a need for more stringent control measures.

Prevalence of Microsporum canis from Pet Cats in Small Animal Hospitals, Chiang Mai, Thailand

Dermatophytosis is a disease caused by dermatophytes, a group of fungi that can cause disease both in humans and animals. The important genera that are pathogenic in animals include Trichophyton and Microsporum. Microsporum canis is an important species because it can cause zoonosis and is commonly found in domestic animals. Cats, which live very close to humans, may expose humans to this pathogen. This research focused on the epidemiology of M. canis found in cats. Hair samples were collected via the Mackenzie technique from cats with and without skin lesions, preliminarily examined with 10% KOH preparation, and cultured for fungal identification. Samples were confirmed with molecular techniques including polymerase chain reaction, gel electrophoresis, and sequencing. Samples were collected from 138 cats located in 93 households, 43 from cats with skin lesions (31.16%) and 95 from cats without skin lesions (68.84%). Eighteen cats with lesions (13.04%) and ten cats without lesions (7.2%) were found to carry M. canis. In eleven of the eighteen cats both with skin lesions and positive for M. canis (61.11%), the pathogen was found both at the site of the lesion and at other sites in the body. Because the pathogen can be found in the hair of cats with and without skin lesions, owners, keepers, veterinarians, and others who come into contact with these animals are at risk of infection if they are not aware or do not take precautions after contact with them.

Optimized isolation of 7,7′-biphyscion starting from Cortinarius rubrophyllus, a chemically unexplored fungal species rich in photosensitizers

Mushrooms such as the dermocyboid Cortinarius rubrophyllus are characterized by strikingly colorful fruiting bodies. The molecular dyes responsible for such colors recently experienced a comeback as photoactive compounds with remarkable photophysical and photobiological properties. One of them—7,7′-biphyscion—is a dimeric anthraquinone that showed promising anticancer effects in the low nanomolar range under blue-light irradiation. Compared to acidic anthraquinones, 7,7′-biphyscion was more efficiently taken up by cells and induced apoptosis after photoactivation. However, seasonal collection of mushrooms producing this compound, low extraction yields, and tricky fungal identification hamper further developments to the clinics. To bypass these limitations, we demonstrate here an alternative approach utilizing a precursor of 7,7′-biphyscion, i.e., the pre-anthraquinone flavomannin-6,6′-dimethyl ether, which is abundant in many species of the subgenus Dermocybe. Controlled oxidation of the crude extract significantly increased the yield of 7,7′-biphyscion by 100%, which eased the isolation process. We also present the mycochemical and photobiological characterization of the yet chemically undescribed species, i.e. C. rubrophyllus. In total, eight pigments (1–8) were isolated, including two new glycosylated anthraquinones (1 and 2). Light-dependent generation of singlet oxygen was detected for the first time for emodin-1-O-β-d-glucopyranoside (3) [photophysical measurement: Φ∆ = 0.11 (CD3OD)]. Furthermore, emodin (7) was characterized as promising compound in the photocytotoxicity assay with EC50-values in the low micromolar range under irradiation against cells of the cancer cell lines AGS, A549, and T24. Graphical abstract

Fungal diversity inhabiting tissues of ebony (Diospyros celebica Bakh.) in urban forest

Eboni (Diospyros celebica Bakh.) is an endemic tree species of Sulawesi Island including Central Sulawesi, West Sulawesi and South Sulawesi. This species is also called fancy wood; its color is black-striped reddish brown, beautiful, and luxurious. In addition, tree growth is influenced by microbes, including fungi. Fungi are heterotrophic and eukaryotic organisms absorbing organic compounds from other organisms. The study aimed to identify ebony-associated fungi in Urban Forestry at the Tamalanrea Campus, Hasanuddin University, Makassar, Indonesia. This study consisted of the isolation stage both direct and dilution methods, the rejuvenation stage, and fungal identification. The study result indicated that there were 60 fungal isolates isolated from the ebony tree tissues, while 19 fungal isolates were isolated on the soil under the ebony stands. The direct isolation-based method was higher in term of number of fungal isolates than the dilution-based method. The isolated fungi belonged to the seven genera, namely Aspergillus, Penicillium, Gliocladium, Trichoderma, Fusarium, Rhizopus, and Phytophthora. Aspergillus and Penicillium was genera dominated both in tree tissues and in the soil under ebony stands.

Mixed-Culture Metagenomics of the Microbes Making Sour Beer

Mixed microbial cultures create sour beers but many brewers do not know which microbes comprise their cultures. The objective of this work was to use deep sequencing to identify microorganisms in sour beers brewed by spontaneous and non-spontaneous methods. Twenty samples were received from brewers, which were processed for microbiome analysis by next generation sequencing. For bacteria, primers were used to amplify the V3-V4 region of the 16S rRNA gene; fungal DNA detection was performed using primers to amplify the entire internal transcribed spacer region. The sequencing results were then used for taxonomy assignment, sample composition, and diversity analyses, as well as nucleotide BLAST searching. We identified 60 genera and 140 species of bacteria, of which the most prevalent were Lactobacillus acetotolerans, Pediococcus damnosus, and Ralstonia picketti/mannitolilytica. In fungal identification, 19 genera and 26 species were found, among which the most common yeasts were Brettanomyces bruxellensis and Saccharomyces cerevisiae. In some cases, genetic material from more than 60 microorganisms was found in a single sample. In conclusion, we were able to determine the microbiomes of various mixed cultures used to produce beer, providing useful information to better understand the sour beer fermentation process and brewing techniques.

Diversity of Pathogenic Fungi and Disease on Vegetable Crops at Polyculture Systems

Vegetables polyculture system is potentially increasing pathogenic fungi diversity because various plant hosts are available. There is no data about patogenic fungi diversity at polyculture vegetable farming in Serang village, District of Karangreja, Purbalingga Regency. This study aimed to determine patogenic fungal diversity and disease percentage caused by the fungi at polyculture vegetable farming in Serang village, District of Karangreja, Purbalingga Regency. This research used purposive random sampling. Infected plants were collected at ten polyculture farming locations and fungal identification was performed at the laboratory. Fungi were identified morphologically based on the signs, symptoms, as well as macroscopic and microscopic characters. The fungi's pathogenity was determined by applying Koch's postulate test. The data were analyzed descriptively through literature comparison. The results showed that seven fungal species were found at polyculture farms in Serang Village. The obtained fungi were Colletotrichum sp., Fusarium sp., Alternaria sp., Septoria sp., Cercospora sp., Botryodiplodia sp., and Nigrospora sp. The lowest damage was 18.24% on tomato fruit infected by Fusarium sp. and the highest was on chili plants which was caused by Colletotrichum sp. The data is the first report for polycuture system. The obtained data has important implication for the management of vegetables farming in Serang Village.

Colletotrichum Species Related To “Nam Dork Mai See Tong” Mango In Central Thailand And Risk Detection of Carbendazim-Resistant Strains

Nam Dork Mai See Tong mango is a deluxe commercial fruit in Thailand, however anthracnose disease becoming a major problem affecting market-driven. Fungicide treatment is the main component in the mango anthracnose management, notwithstanding the appearance of fungal-resistance has been become a factor of fungicide limition and can affect to increasingly higher costs. Thirty-two isolates of the Colletotrichum species complex were obtained from anthracnose-diseased mango cv. Nam Dork Mai See Tong in 3 provinces of Thailand. Fungal identification based on morphological and molecular markers was investigated. All isolates were divided into 3 species: C. asianum , C. gloeosporioides and C. siamense . Pathogenicity tests revealed that all 3 species caused symptoms on artificially unwounded mango fruits and leaves. Only C. gloeosporioides have previously been reported on mango, while C. asianum and C. siamense were first reports associated with mango in Thailand. The responsiveness of Colletotrichum isolates to carbendazim was evaluated using an MIC assay. Nine isolates of C. asianum were highly resistant, while 5 and 8 isolates of C. gloeosporioides were resistant (R) and highly resistant (HR), respectively. Moreover, all isolates of C. siamense were HR. Mutations were detected by PCR of a partial sequence of the β-tubulin gene. The sequence of β-tubulin gene in HR strains showed a single nucleotide transversion of adenine to cytosine, resulting in a substitution at codon 198. However, R strains were found to have only an amino acid change at codon 200. Additionally, PCR-RFLP was applied as a rapid technique to detect carbendazim resistance. The results demonstrated that only the Bsh 1236I restriction enzyme generated two bands (200 and 300 bp) in the highly resistant strain, but these bands were absent in the sensitive (S) and R strains. Hence, PCR-RFLP technique showed the potential to specifically detect benzimidazole fungicide resistance in 3 species of Colletotrichum. Moreover, this technique is accessible and feasible for laboratory assessment

Mixed culture metagenomics of the microbes making sour beer

Mixed microbial cultures create sour beers, but many brewers do not know which microbes comprise their cultures. The objective of this work was to use deep sequencing to identify mi-croorganisms in sour beers brewed by spontaneous and non-spontaneous methods. Twenty samples were received from brewers, which were processed for microbiome analysis by next generation sequencing. For bacteria, primers were used to amplify the V3-V4 region of the 16S rRNA gene; fungal DNA detection was performed using primers to amplify the entire internal transcribed spacer region. The sequencing results were then used for taxonomy assignment, sample composition, and diversity analyses, as well as nucleotide BLAST searching. We identi-fied 60 genera and 140 species of bacteria, of which the most prevalent were Lactobacillus acetotol-erans, Pediococcus damnosus, and Ralstonia picketti/mannitolilytica. In fungal identification, 19 genera and 26 species were found, among which the most common yeasts were Brettanomyces bruxellen-sis and Saccharomyces cerevisiae. In some cases, genetic material from more than 60 microorgan-isms was found in a single sample. In conclusion, we were able to determine the microbiomes of various mixed cultures used to produce beer, providing useful information to better understand the sour beer fermentation process and brewing techniques.

Analysis of Microbiota and Mycobiota in Fungal Ball Rhinosinusitis: Specific Interaction between Aspergillus fumigatus and Haemophilus influenza?

Fungal ball (FB) rhinosinusitis (RS) is the main type of non-invasive fungal RS. Despite positive direct examination (DE) of biopsies, culture remains negative in more than 60% of cases. The aim of the study was to evaluate the performance/efficacy of targeted metagenomics (TM) to analyze microbiota and mycobiota in FB and find microbial associations. Forty-five sinus biopsies from patients who underwent surgery for chronic RS were included. After DE and culture, DNA was extracted, then fungal ITS1–ITS2 and bacterial V3–V4 16S rDNA loci were sequenced (MiSeqTM Illumina). Operational taxonomic units (OTUs) were defined via QIIME and assigned to SILVA (16S) and UNITE (ITS) databases. Statistical analyses were performed using SHAMAN. Thirty-eight patients had FB and seven had non-fungal rhinosinusitis (NFRS). DE and culture of FB were positive for fungi in 97.3 and 31.6% of patients, respectively. TM analysis of the 38 FB yielded more than one fungal genus in 100% of cases, with Aspergillus in 89.5% (34/38). Haemophilus was over-represented in FB with >1000 reads/sample in 47.3% (18/38) compared to NFRS (p < 0.001). TM allowed fungal identification in biopsies with negative culture. Haemophilus was associated with FB. Pathogenesis could result from fungi–bacteria interactions in a mixed biofilm-like structure.