Natural occurrence of aflatoxin, aflatoxigenic and nonaflatoxigenic Aspergillus flavus in groundnut seeds across India

A survey across different agro-climatic regions of India was done and 38 groundnut seed samples were collected from various sources. Upon analysis, all samples were found infected with Aspergillus flavus ranging from 2 to 50% incidence with aflatoxin content of 0.0 to 28 ppb. Greenhouse studies revealed no correlation between incidence of A. flavus and aflatoxin content on seedling emergence, root length, shoot length and dry weight. Seeds were predominantly contaminated with aflatoxin B1 followed by aflatoxin B2. Among the tested A. flavus isolates, 31 were found aflatoxigenic and seven were nonaflatoxigenic when analyzed through cultural, thin layer chromatography, competitive direct enzyme linked immunosorbent assay and multiplex polymerase chain reaction. Present study reveals the current scenario of aflatoxin contamination, and aflatoxigenic and non-aflatoxigenic fungal infection in groundnut seeds collected across India.Keywords: Polymerase chain reaction (PCR), Aspergillus flavus, aflatoxin, enzyme-linked immuno sorbent assay (ELISA), groundnutAfrican Journal of Biotechnology Vol. 12(19), pp. 2587-259

Neuroprotective Effects of Bikaverin on H2O2-Induced Oxidative Stress Mediated Neuronal Damage in SH-Sy5y Cell Line

The generation of free radicals and oxidative stress has been linked to several neurodegenerative diseases including Parkinson's disease, Alzheimer's disease, Huntington's disease, and Amyotrophic lateral sclerosis. The use of free radical scavenging molecules for the reduction of intracellular reactive oxygen species is one of the strategies used in the clinical management of neurodegeneration. Fungal secondary metabolism is a rich source of novel molecules with potential bioactivity. In the current study, bikaverin was extracted from Fusarium oxysporum f. sp. lycopersici and its structural characterization was carried out. Further, we explored the protective effects of bikaverin on oxidative stress and its anti-apoptotic mechanism to attenuate H2O2-induced neurotoxicity using human neuroblastoma SH-SY5Y cells. Our results elucidate that pretreatment of neurons with bikaverin attenuates the mitochondrial and plasma membrane damage induced by 100 µM H2O2 to 82 and 26 % as evidenced by MTT and LDH assays. H2O2 induced depletion of antioxidant enzyme status was also replenished by bikaverin which was confirmed by Realtime Quantitative PCR analysis of SOD and CAT genes. Bikaverin pretreatment efficiently potentiated the H2O2-induced neuronal markers, such as BDNF, TH, and AADC expression, which orchestrate the neuronal damage of the cell. The H2O2-induced damage to cells, nuclear, and mitochondrial integrity was also restored by bikaverin. Bikaverin could be developed as a preventive agent against neurodegeneration and as an alternative to some of the toxic synthetic antioxidants

Fungal Planet description sheets: 1436–1477

Novel species of fungi described in this study include those from various countries as follows: Argentina, Colletotrichum araujiae on leaves, stems and fruits of Araujia hortorum. Australia, Agaricus pateritonsus on soil, Curvularia fraserae on dying leaf of Bothriochloa insculpta, Curvularia millisiae from yellowing leaf tips of Cyperus aromaticus, Marasmius brunneolorobustus on well-rotted wood, Nigrospora cooperae from necrotic leaf of Heteropogon contortus, Penicillium tealii from the body of a dead spider, Pseudocercospora robertsiorum from leaf spots of Senna tora, Talaromyces atkinsoniae from gills of Marasmius crinis-equi and Zasmidium pearceae from leaf spots of Smilax glyciphylla. Brazil, Preussia bezerrensis from air. Chile, Paraconiothyrium kelleni from the rhizosphere of Fragaria chiloensis subsp. chiloensis f. chiloensis. Finland, Inocybe udicola on soil in mixed forest with Betula pendula, Populus tremula, Picea abies and Alnus incana. France, Myrmecridium normannianum on dead culm of unidentified Poaceae. Germany, Vexillomyces fraxinicola from symptomless stem wood of Fraxinus excelsior. India, Diaporthe limoniae on infected fruit of Limonia acidissima, Didymella naikii on leaves of Cajanus cajan, and Fulvifomes mangroviensis on basal trunk of Aegiceras corniculatum. Indonesia, Penicillium ezekielii from Zea mays kernels. Namibia, Neocamarosporium calicoremae and Neocladosporium calicoremae on stems of Calicorema capitata, and Pleiochaeta adenolobi on symptomatic leaves of Adenolobus pechuelii. Netherlands, Chalara pteridii on stems of Pteridium aquilinum, Neomackenziella juncicola (incl. Neomackenziella gen. nov.) and Sporidesmiella junci from dead culms of Juncus effusus. Pakistan, Inocybe longistipitata on soil in a Quercus forest. Poland, Phytophthora viadrina from rhizosphere soil of Quercus robur, and Septoria krystynae on leaf spots of Viscum album. Portugal (Azores), Acrogenospora stellata on dead wood or bark. South Africa, Phyllactinia greyiae on leaves of Greyia sutherlandii and Punctelia anae on bark of Vachellia karroo. Spain, Anteaglonium lusitanicum on decaying wood of Prunus lusitanica subsp. lusitanica, Hawksworthiomyces riparius from fluvial sediments, Lophiostoma carabassense endophytic in roots of Limbarda crithmoides, and Tuber mohedanoi from calcareus soils. Spain (Canary Islands), Mycena laurisilvae on stumps and woody debris. Sweden, Elaphomyces geminus from soil under Quercus robur. Thailand, Lactifluus chiangraiensis on soil under Pinus merkusii, Lactifluus nakhonphanomensis and Xerocomus sisongkhramensis on soil under Dipterocarpus trees. Ukraine, Valsonectria robiniae on dead twigs of Robinia hispida. USA, Spiralomyces americanus (incl. Spiralomyces gen. nov.) from office air. Morphological and culture characteristics are supported by DNA barcodes

Fungal Planet description sheets: 1478-1549

Novel species of fungi described in this study include those from various countries as follows: Australia, Aschersonia mackerrasiae on whitefly, Cladosporium corticola on bark of Melaleuca quinquenervia, Penicillium nudgee from soil under Melaleuca quinquenervia, Pseudocercospora blackwoodiae on leaf spot of Persoonia falcata, and Pseudocercospora dalyelliae on leaf spot of Senna alata. Bolivia, Aspicilia lutzoniana on fully submersed siliceous schist in high-mountain streams, and Niesslia parviseta on the lower part and apothecial discs of Erioderma barbellatum onatwig. Brazil, Cyathus bonsai on decaying wood, Geastrum albofibrosum from moist soil with leaf litter, Laetiporus pratigiensis on a trunk of a living unknown hardwood tree species, and Scytalidium synnematicum on dead twigs of unidentified plant. Bulgaria, Amanita abscondita on sandy soil in a plantation of Quercus suber. Canada, Penicillium acericola on dead bark of Acer saccharum, and Penicillium corticola on dead bark of Acer saccharum. China, Colletotrichum qingyuanense on fruit lesion of Capsicum annuum. Denmark, Helminthosphaeria leptospora on corticioid Neohypochnicium cremicolor. Ecuador (Galapagos), Phaeosphaeria scalesiae on Scalesia sp. Finland, Inocybe jacobssonii on calcareouss oils in dry forests and park habitats. France, Cortinarius rufomyrrheus on sandy soil under Pinus pinaster, and Periconia neominutissima on leaves of Poaceae. India, Coprinopsis fragilis on decaying bark of logs, Filoboletus keralensis on unidentified woody substrate, Penicillium sankaranii from soil, Physisporinus tamilnaduensis on the trunk of Azadirachta indica, and Poronia nagaraholensis on elephant dung. Iran, Neosetophoma fic on infected leaves of Ficus elastica. Israel, Cnidariophoma eilatica (incl. Cnidariophoma gen. nov.) from Stylophora pistillata. Italy, Lyophyllum obscurum on acidic soil. Namibia, Aureobasidium faidherbiae on dead leaf of Faidherbia albida, and Aureobasidium welwitschiae on dead leaves of Welwitschia mirabilis. Netherlands, Gaeumannomycella caricigena on dead culms of Carex elongata, Houtenomyces caricicola (incl. Houtenomyces gen. nov.) on culms of Carex disticha, Neodacampia ulmea (incl. Neodacampia gen. nov.) on branch of Ulmus laevis, Niesslia phragmiticola on dead standing culms of Phragmites australis, Pseudopyricularia caricicola on culms of Carex disticha, and Rhodoveronaea nieuwwulvenica on dead bamboo sticks. Norway, Arrhenia similis half-buried and moss-covered pieces of rotting wood in grass-grownpath. Pakistan, Mallocybe ahmadii on soil. Poland, Beskidomyces laricis (incl. Beskidomyces gen. nov.) from resin of Larix decidua ssp. polonica, Lapidomyces epipinicola from sooty mould community on Pinus nigra, and Leptographium granulatum from a gallery of Dendroctonus micans on Picea abies. Portugal, Geoglossum azoricum on mossy areas of laurel forest areas planted with Cryptomeria japonica, and Lunasporangiospora lusitanica from a biofilm covering a bio deteriorated limestone wall. Qatar, Alternaria halotolerans from hypersaline sea water, and Alternaria qatarensis from water sample collected from hypersaline lagoon. South Africa, Alfaria thamnochorti on culm of Thamnochortus fraternus, Knufia aloeicola on Aloe gariepensis, Muriseptatomyces restionacearum (incl.Muriseptatomyces gen. nov.) on culms of Restionaceae, Neocladosporium arctotis on nest of cases of bagworm moths(Lepidoptera, Psychidae) on Arctotis auriculata, Neodevriesia scadoxi on leaves of Scadoxus puniceus, Paraloratospora schoenoplecti on stems of Schoenoplectus lacustris, Tulasnella epidendrea from the roots of Epidendrum × obrienianum, and Xenoidriella cinnamomi (incl. Xenoidriella gen. nov.) on leaf of Cinnamomum camphora. South Korea, Lemonniera fraxinea on decaying leaves of Fraxinus sp. frompond. Spain, Atheniella lauri on the bark of fallen trees of Laurus nobilis, Halocryptovalsa endophytica from surface-sterilised, asymptomatic roots of Salicornia patula, Inocybe amygdaliolens on soil in mixed forest, Inocybe pityusarum on calcareous soil in mixed forest, Inocybe roseobulbipes on acidic soils, Neonectria borealis from roots of Vitis berlandieri × Vitis rupestris, Sympoventuria eucalyptorum on leaves of Eucalyptus sp., and Tuber conchae fromsoil. Sweden, Inocybe bidumensis on calcareous soil. Thailand, Cordyceps sandindaengensis on Lepidoptera pupa, buried in soil, Ophiocordyceps kuchinaraiensis on Coleoptera larva, buried in soil, and Samsoniella winandae on Lepidoptera pupa, buriedinsoil. Taiwan region (China), Neophaeosphaeria livistonae on dead leaf of Livistona rotundifolia. Türkiye, Melanogaster anatolicus on clay loamy soils. UK, Basingstokeomyces allii (incl. Basingstokeomyces gen. nov.) on leaves of Allium schoenoprasum. Ukraine, Xenosphaeropsis corni on recently dead stem of Cornus alba. USA, Nothotrichosporon aquaticum (incl. Nothotrichosporon gen. nov.) from water, and Periconia philadelphiana from swab of coil surface. Morphological and culture characteristics for these new taxa are supported by DNA barcodes.The work of P.W. Crous and colleagues benefitted from funding by the European Union’s Horizon 2020 research and innovation program (RISE) under the Marie Skłodowska-Curie grant agreement No. 101008129, project acronym ‘Mycobiomics’, and the Dutch NWO Roadmap grant agreement No. 2020/ENW/00901156, project ‘Netherlands Infrastructure for Ecosystem and Biodiversity Analysis – Authoritative and Rapid Identification System for Essential biodiversity information’(acronym NIEBAARISE). G. Gulden, B. Rian and I. Saar thank K. Bendiksen at the fungarium and G. Marthinsen at NorBol, both Natural History Museum, University of Oslo for valuable help with the collections, and the sequencing of our finds of A. similis from 2022. Sincere thanks to A. Voitk for assistance with the colour plate and review of the manuscript. I. Saar was supported by the Estonian Research Council (grant PRG1170). P. Rodriguez-Flakus and co-authors are greatly indebted to their colleagues and all staff of the Herbario Nacional de Bolivia, Instituto de Ecología, Universidad Mayor de SanAndrés, La Paz, for their generous long-term cooperation. Their research was financially supported by the National Science Centre (NCN) in Poland (grants numbers 2018/02/X/NZ8/02362 and 2021/43/B/NZ8/02902). Y.P. Tan and colleagues thank M.K. Schutze (Department of Agriculture and Fisheries, Queensland, Australia) for determining the identity of the insect hosts for Aschersonia mackerrasiae. The Australian Biological Resources Study funded the project that led to the discovery of Aschersonia mackerrasiae. K.G.G. Ganga acknowledges support from the University Grants Commission (UGC), India, in the form of a UGC research fellowship (Ref No. 20/12/2015(ii) EU-V), and the authorities of the University of Calicut for providing facilities to conduct this study. S. Mahadevakumar acknowledges the Director, KSCSTE - Kerala Forest Research Institute and Head of Office, Botanical Survey of India,Andaman and Nicobar Regional Centre, Port Blair for the necessary support and M. Madappa, Department of Studies in Botany, University of Mysore for technical assistance. A.R. Podile thanks the Department of Science and Technology, Govt. of India for the JC Bose Fellowship (Grant No. JCB/2017/000053) & MoE and IOE-Directorate-UOH for project (Grant No.UOH-IOE-RC3-21-065). Financial support was provided to R. de L. Oliveira and K.D. Barbosa by the Coordenação deAperfeiçoamento de Pessoal de Nível Superior - Brazil (CAPES) – Finance code 001, and to I.G. Baseia and M.P. Martín by the National Council for Scientific and Technological Development (CNPq) under CNPq-Universal 2016 (409960/2016-0) and CNPq-visiting researcher (407474/2013-7). E. Larsson acknowledges the Swedish Taxonomy Initiative, SLU Artdatabanken, Uppsala, Sweden. H.Y. Mun and J. Goh were supported by a grant from the Nakdonggang National Institute of Biological Resources (NNIBR), funded by the Ministry of Environment (MOE) of the Republic of Korea (NNIBR202301106). J. Trovão and colleagues were financed by FEDER - Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020 - Operational Programme for Competitiveness and Internationalisation (POCI), and by Portuguese funds through FCT- Fundação para a Ciência e a Tecnologia in the framework of the project POCI-01-0145-FEDER-PTDC/EPH-PAT/3345/ 2014. Their research was carried out at the R & D Unit Centre for Functional Ecology – Science for People and the Planet (CFE), with reference UIDB/04004/2020, financed by FCT/MCTES through national funds (PIDDAC). João Trovão was supported by POCH - Programa Operacional Capital Humano (co-funding by the European Social Fund and national funding by MCTES), through a ‘FCT- Fundação para a Ciência e Tecnologia’ PhD research grant (SFRH/ BD/132523/2017). O. Kaygusuz and colleagues thank the Research Fund of the Isparta University ofApplied Sciences for their financial support under the project number 2021-ILK1-0155. They also thank N. Sánchez Biezma of the Department of Drawing and Scientific Photography at the Alcalá University for his help in the digital preparation of the photographs. The research of M. Spetik and co-authors was supported by project No. IGAZF/2021-SI1003. V. Darmostuk and colleagues acknowledge our colleagues and all staff of the Herbario Nacional de Bolivia, Instituto de Ecología, Universidad Mayor de San Andrés, La Paz, for their generous long-term cooperation. They would also like to thank the SERNAP (http://sernap.gob.bo), and all protected areas staff, for providing permits for scientific studies, as well as their assistance and logistical support during the field works. This research was financially supported by the National Science Centre (NCN) in Poland (grant number DEC-2013/11/D/NZ8/ 03274). M. Kaliyaperumal and co-authors thank the Centre of Advanced Studies in Botany, University of Madras for the laboratory facilities. M. Kaliyaperumal thanks the Extramural Research-SERB, DST (EMR/2016/003078), Government of India, for financial assistance. M. Kaliyaperumal and K. Kezo thanks RUSA 2.0 (Theme-1, Group-1/2021/49) for providing a grant. M. Shivannegowda and colleagues thank C.R. Santhosh, Department of Studies in Microbiology, University of Mysore, Manasagangotri, Mysuru for technical support. They also thank K.R. Sridhar, Mangalore University, Karnataka, India and S.S.N. Maharachchikumbura, University of Electronic Science and Technology of China, Chengdu for their support and helping with technical inputs. The study of G.G. Barreto and co-authors was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brazil (CAPES - Finance Code 001), and the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq - Proc. 131503/2019-7; Proc. 312984/2018-9); the authors also thank to Programa de Pós-Graduação em Botânica – PPGBOT. L.F.P. Gusmão thanks to Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for a research grant. T. Nkomo and colleagues thank the National Research Foundation of SouthAfrica for funding this study, with additional funding from the Forestry and Agricultural Biotechnology Institute and the University of Pretoria. G. Delgado is grateful to W. Colbert and S. Ward (Eurofins Built Environment) for continual encouragement and provision of laboratory facilities. J.G. Maciá-Vicente acknowledges support from the Landes-Offensive zur Entwicklung Wissenschaftlich-ökonomischer Exzellenz (LOEWE) of the state of Hesse within the framework of the Cluster for Integrative Fungal Research (IPF) of Goethe University Frankfurt. F. Esteve-Raventós and colleagues acknowledge P. Juste and J.C. Campos for the loan of some collections for study and N. Subervielle and L. Hugot (Conservatoire Botanique National de Corse, Office de l’Environnement de la Corse, Corti) for their assistance. They also acknowledge the Balearic Mycology Group (FCB) for their permanent help in the search for collections in the Balearic Islands, and Y. Turégano for obtaining some of the sequences presented here, and L. Parra for his suggestions and help on nomenclatural issues. S. Mongkolsamrit and colleagues were financially supported by the Platform Technology Management Section, National Centre for Genetic Engineering and Biotechnology (BIOTEC), Project Grant No. P19-50231. S. De la Peña-Lastra and colleagues thank the Atlantic Islands National Maritime-Terrestrial Park authorities and guards. A. Mateos and co-authors would like to thank Secretaria Regional doAmbiente eAlterações Climáticas Açores for the permission granted for the sampling (Licença nº 16/2021/ DRAAC). To the ECOTOX group for co-funding the trip. J. Mack & D.P. Overy were funded byAgriculture &Agri-Food Canada (Project ID#002272: Fungal and Bacterial Biosystematics-bridging taxonomy and “omics” technology in agricultural research and regulation) and are grateful for molecular sequencing support from the Molecular Technologies Laboratory (MTL) at the Ottawa Research & Development Centre of Agriculture & Agri-Food Canada. The study of P. Czachura was funded by the National Science Centre, Poland, under the project 2019/35/N/NZ9/04173. The study of M. Piątek and coauthors was funded by the National Science Centre, Poland, under the project 2017/27/B/NZ9/02902. O. Yarden and L. Granit were funded by the Israel Science Foundation (grant number 888/19). H. Taşkın and colleagues received support from the BulgarianAcademy of Sciences and the Scientific and Technological Research Council of Türkiye (Bilateral grant agreement between BAS and TÜBİTAK, project number 118Z640). The authors would also like to thank S. Şahin (İzmir, Türkiye) for conveying one of the localities of A. abscondita. Andrew Miller would like to thank the Roy J. Carver Biotechnology Center at the University of Illinois for Sanger sequencing. E.R. Osieck thanks Staatsbosbeheer for permission to collect fungi in Nieuw Wulven, in the Netherlands. P. van ‘t Hof and co-authors thank the Galapagos Genetic Barcode project supported by UK Research and Innovation, Global Challenges Research Fund, Newton Fund, University of Exeter, Galapagos Science Center, Universidad San Francisco de Quito, Galapagos Conservation Trust, and Biosecurity Agency of Galapagos (ABG).Peer reviewe

Cytotoxic effects of oosporein isolated from endophytic fungus cochliobolus kusanoi

In the present study, oosporein, a fungal toxic secondary metabolite known to be a toxic agent causing chronic disorders in animals, was isolated from fungus Cochliobolus kusanoi of Nerium oleander L. Toxic effects of oosporein and the possible mechanisms of cytotoxicity as well as the role of oxidative stress in cytotoxicity to Madin-Darby canine kidney kidney cells and RAW 264.7 splene cells were evaluated in vitro. Also to know the possible in vivo toxic effects of oosporein on kidney and spleen, Balb/C mouse were treated with different concentrations of oosporein ranging from 20 to 200 mu M). After 24 h of exposure histopathological observations were made to know the effects of oosporein on target organs. Oosporein induced elevated levels of reactive oxygen species (ROS) generation and high levels of malondialdehyde, loss of mitochondrial membrane potential, induced glutathione hydroxylase (GSH) production was observed in a dose depended manner. Effects oosporein on chromosomal DNA damage was assessed by Comet assay, and increase in DNA damage were observed in both the studied cell lines by increasing the oosporein concentration. Further, oosporein treatment to studied cell lines indicated significant suppression of oxidative stress related gene (Superoxide dismutasel and Catalase) expression, and increased levels of mRNA expression in apoptosis or oxidative stress inducing genes HSP70, Caspase3, Caspase6, and Caspase9 as measured by quantitative real time-PCR assay. Histopathological examination of oosporein treated mouse kidney and splenocytes further revealed that, oosporein treated target mouse tissues were significantly damaged with that of untreated sam control mice and these effects were in directly proportional to the the toxin dose. Results of the present study reveals that, ROS is the principle event prompting increased oosporein toxicity in studied in vivio and in vitro animal models. The high previlance of these fungi in temperate climates further warrants the need of safe food grain storage and processing practices to control the toxic effects of oosporein to humans and live stock

A novel igy-aptamer hybrid system for cost-effective detection of seb and its evaluation on food and clinical samples

In the present study, we introduce a novel hybrid sandwich-ALISA employing chicken IgY and ssDNA aptamers for the detection of staphylococcal enterotoxin B (SEB). Cloning, expression and purification of the full length recombinant SEB was carried out. Anti-SEB IgY antibodies generated by immunizing white leg-horn chickens with purified recombinant SEB protein and were purified from the immunized egg yolk. Simultaneously, ssDNA aptamers specific to the toxin were prepared by SELEX method on microtiter well plates. The sensitivity levels of both probe molecules i.e., IgY and ssDNA aptamers were evaluated. We observed that the aptamer at 250 ngmL(-1) concentration could detect the target antigen at 50 ngmL(-1) and the IgY antibodies at 250 ngmL(-1), could able to detect 100 ngmL(-1) antigen. We further combined both the probes to prepare a hybrid sandwich aptamer linked immune sorbent assay (ALISA) wherein the IgY as capturing molecule and biotinylated aptamer as revealing probe. Limit of detection (LOD) for the developed method was determined as 50 ngmL(-1). Further, developed method was evaluated with artificially SEB spiked milk and natural samples and obtained results were validated with PCR. In conclusion, developed ALISA method may provide cost-effective and robust detection of SEB from food and environmental samples