New species in Aciculosporium, Shimizuomyces and a new genus Morakotia associated with plants in Clavicipitaceae from Thailand.

Three new fungal species in the Clavicipitaceae (Hypocreales, Ascomycota) associated with plants were collected in Thailand. Morphological characterisation and phylogenetic analyses based on multi-locus sequences of LSU, RPB1 and TEF1 showed that two species belong to Aciculosporium and Shimizuomyces. Morakotia occupies a unique clade and is proposed as a novel genus in Clavicipitaceae. Shimizuomyces cinereus and Morakotia fusca share the morphological characteristic of having cylindrical to clavate stromata arising from seeds. Aciculosporium siamense produces perithecial plates and occurs on a leaf sheath of an unknown panicoid grass.

Fungal pathogens occurring on Orthopterida in Thailand.

Two new fungal genera and six species occurring on insects in the orders Orthoptera and Phasmatodea (superorder Orthopterida) were discovered that are distributed across three families in the Hypocreales. Sixty-seven sequences generated in this study were used in a multi-locus phylogenetic study comprising SSU, LSU, TEF, RPB1 and RPB2 together with the nuclear intergenic region (IGR). These new taxa are introduced as Metarhizium gryllidicola, M. phasmatodeae, Neotorrubiella chinghridicola, Ophiocordyceps kobayasii, O. krachonicola and Petchia siamensis. Petchia siamensis shows resemblance to Cordyceps mantidicola by infecting egg cases (ootheca) of praying mantis (Mantidae) and having obovoid perithecial heads but differs in the size of its perithecia and ascospore shape. Two new species in the Metarhizium cluster belonging to the M. anisopliae complex are described that differ from known species with respect to phialide size, conidia and host. Neotorrubiella chinghridicola resembles Torrubiella in the absence of a stipe and can be distinguished by the production of whole ascospores, which are not commonly found in Torrubiella (except in Torrubiella hemipterigena, which produces multiseptate, whole ascospores). Ophiocordyceps krachonicola is pathogenic to mole crickets and shows resemblance to O. nigrella, O. ravenelii and O. barnesii in having darkly pigmented stromata. Ophiocordyceps kobayasii occurs on small crickets, and is the phylogenetic sister species of taxa in the 'sphecocephala' clade.

Revisiting Metarhizium and the description of new species from Thailand.

Over the last two decades the molecular phylogeny and classification of Metarhizium has been widely studied. Despite these efforts to understand this enigmatic genus, the basal lineages in Metarhizium are still poorly resolved. In this study, a phylogenetic framework is reconstructed for the Clavicipitaceae focusing on Metarhizium through increased taxon-sampling using five genomic loci (SSU, LSU, tef, rpb1, rpb2) and the barcode marker ITS rDNA. Multi-gene phylogenetic analyses and morphological characterisation of green-spored entomopathogenic Metarhizium isolates from Thailand and soil isolates of M. carneum and M. marquandii reveal their ecological, genetic and species diversity. Nineteen new species are recognised in the Metarhizium clade with narrow host ranges: two new species are found in the M. anisopliae complex - M. clavatum on Coleoptera larvae and M. sulphureum on Lepidoptera larvae; four new species are found in the M. flavoviride complex - M. biotecense and M. fusoideum on brown plant hoppers (Hemiptera), M. culicidarum on mosquitoes, M. nornnoi on Lepidoptera larvae; three new species M. megapomponiae, M. cicadae, M. niveum occur on cicadas; five new species M. candelabrum, M. cercopidarum, M. ellipsoideum, M. huainamdangense M. ovoidosporum occur on planthoppers, leafhoppers and froghoppers (Hemiptera); one new species M. eburneum on Lepidoptera pupae; and four new species M. phuwiangense, M. purpureum, M. purpureonigrum, M. flavum on Coleoptera . Of these 19 new species, seven produce a sexual morph (M. clavatum, M. eburneum, M. flavum, M. phuwiangense, M. purpureonigrum, M. purpureum, and M. sulphureum) and asexual morphs are found in the remaining new species and also in M. sulphureum, M. purpureonigrum and M. purpureum. Metarhizium blattodeae, M. koreanum and M. viridulum are new records for Thailand. An alternative neotype for Metarhizium anisopliae is proposed based on multi-gene and 5'tef analyses showing that CBS 130.71 from Ukraine is more suitable, being from a much closer geographical location to Metchnikoff's Metarhizium anisopliae. This isolate is distinct from the neotype of Metarhizium anisopliae var. anisopliae proposed by M. Tulloch from Ethiopia (ARSEF 7487). Six new genera are established for monophyletic clades subtending the core Metarhizium clade, including Keithomyces, Marquandomyces, Papiliomyces, Purpureomyces, Sungia, and Yosiokobayasia. Metarhizium carneum, M. aciculare, and M. neogunnii are combined in Keithomyces and one new combination for M. marquandii in Marquandomyces is proposed. Purpureomyces is introduced for species producing purple stromata including a new combination for M. khaoyaiense and two new species P. maesotensis and P. pyriformis. Papiliomyces contains two new combinations for M. liangshanense and Metacordyceps shibinensis. The genus Sungia is proposed for the Korean species M. yongmunense on Lepidoptera pupa and Yosiokobayasia for the Japanese species M. kusanagiense also on Lepidoptera pupa. A synoptic and dichotomous key to the accepted taxa is provided together with tables listing distinguishing morphological characters between species, host preferences, and geography.

Prevalence and risk factors for intestinal carriage of CTX-M-type ESBLs in Enterobacteriaceae from a Thai community.

The incidence of infections caused by antimicrobial-resistant Enterobacteriaceae in Thailand is increasing and human intestinal flora is an important reservoir for these organisms. This study was carried out to determine the intestinal carriage of bla CTX-M extended spectrum ß-lactamase-positive Enterobacteriaceae (ESBL + E) and AmpC-positive Enterobacteriaceae in a community setting in Northern Thailand, and to identify potential risk factors for carriage. A total of 307 fecal samples were collected from healthy volunteers in Phitsanulok province, and cefotaxime-resistant Enterobacteriaceae (CtxRE) were isolated using selective media. Polymerase chain reaction (PCR) was used to detect ESBL and AmpC genes. Risk factors were analyzed using multiple logistic regression. Genotyping was performed by multilocus sequence typing (MLST) analysis. Two hundred ninety-one CtxRE isolates were obtained and Escherichia coli was the predominant organism (66.3%). The intestinal carriage rates of bla CTX-M ESBL + E and AmpC-positive Enterobacteriaceae were 52.1% and 6.2%, respectively. Comparative levels of bla CTX-M group 1 and bla CTX-M group 9 were found while bla CMY-2 was the predominant genotype among AmpC genes. Co-existence of two ß-lactamase genes in a single isolate was found in 6.5% of isolates. Consumption of undercooked meat was strongly associated with intestinal carriage of bla CTX-M ESBL + E (p = 0.003, OR = 2.133, 95% CI = 1.289-3.530). Phylogenetic grouping and MLST analysis of E. coli isolates revealed the presence of E. coli B2-ST131 (n = 8). Of these, seven carried bla CTX-M-group 9 and 1 carried bla CMY-2. Our results suggest that residents in Thailand are at high risk for developing endogenous infections caused by antibiotic-resistant Enterobacteriaceae.