Occurrence and identification of Penicillium and Talaromyces species from beach sand

Aims@#Penicillium and Talaromyces were among the species of microfungi that inhabit beach sand in Batu Ferringhi Beach, Penang Island, Malaysia. Previously, Talaromyces was described as the sexual stage of Penicillium, but both are now accepted as separate genera based on molecular phylogeny. The aim of the present study was to identify species of Penicillium and Talaromyces that are present in beach sand in Malaysia.@*Methodology and results@#Species identities were confirmed according to similarities of the internal transcribed spacer regions and β-tubulin gene sequences and a phylogenetic analysis based on both regions/gene. Nine Penicillium spp. were identified as P. georgiense, P. chermesinum, P. pimiteouiense, P. citrinum, P. oxalicum, P. daleae, P. rolfsii and Penicillium sp. and the four Talaromyces spp. were T. siamense, T. atroroseus, T. minioluteus and T. fusiformis.@*Conclusion, significance and impact of study@#These findings showed that beach sand harboured a variety of Penicillium and Talaromyces species. The occurrence of Penicillium and Talaromyces in beach sands is associated with the organic matter in the sand, which provides suitable substrates and nutrient sources. Due to this, beach sand might harbour many potentially pathogenic or opportunistic species that may pose a health concern to immunocompromised individuals.

Statistical optimization of cellulases by Talaromyces thermophilus utilizing Saccharum spontaneum, a novel substrate

BACKGROUND: At present, cellulases are the most important enzymes worldwide, and their demand has been increasing in the industrial sector owing to their notable hydrolysis capability. RESULTS: In the present study, contrary to conventional techniques, three physical parameters were statistically optimized for the production of cellulase by thermophilic fungi by using response surface methodology (RSM). Among all the tested thermophilic strains, the best cellulase producing fungus was identified as Talaromyces thermophilus ­ both morphologically and molecularly through 5.8S/ITS rDNA sequencing. The central composite design (CCD) was used to evaluate the interactive effect of the significant factors. The CCD was applied by considering incubation period, pH, and temperature as the model factors for the present investigation. A second-order quadratic model and response surface method revealed that the independent variables including pH 6, temperature 50 C, and incubation period 72 h significantly influenced the production of cellulases. The analysis of variance (ANOVA) indicated that the established model was significant (P 0.05) and showed the high adequacy of the model. The actual and predicted values of CMCase and FPase activity showed good agreement with each other and also confirmed the validity of the designed model. CONCLUSIONS: We believe the present findings to be the first report on cellulase production by exploiting Kans grass (Saccharum spontaneum) as a substrate through response surface methodology by using thermophilic fungus, Talaromyces thermophilus.

Moringa straw as cellulase production inducer and cellulolytic fungi source / Residuo de moringa como inductor de la producción de celulasas y como fuente de hongos celulolíticos

Abstract Currently, the valorization of agroindustrial waste is of great interest. Moringa oleifera is a multipurpose tree whose softwood residues could be used as raw material for low-cost cellulase production. The aim of this study was to isolate, identify, and characterize microorganisms with cellulolytic activity in different carbon sources. We isolated and puri-fied 42 microorganisms from M. oleifera biomass. Fungi presenting the largest hydrolytic halos in carboxymethylcellulose as a substrate were molecularly identified as Penicillium funiculosum (FG1), Fusarium verticillioides (FG3) and Cladosporium cladosporioides (FC2). The ability of these fungal strains to break down cellulose was assessed in a submerged fermentation using either amorphous CMC or crystalline form (Avicel). P. funiculosum and C. cladosporioides displayed similar endoglucanase (606 U/l) and exoglucanase (205 U/l) activities in the Avicel-containing medium, whereas F. verticillioides showed the highest level of p-glucosidase activity (664 U/l) in the carboxymethylcellulose medium. In addition, the effect of three culture media (A, B, and C) on cellulase production was evaluated in P. funiculosum using moringa straw as a carbon source. The results showed a volumetric productivity improvement of cellulases that was 2.77-, 8.26-, and 2.30-fold higher for endoglucanase, exoglucanase and p-glucosidase, respectively when medium C containing moringa straw was used as a carbon source. The enzymatic extracts produced by these fungi have biotechnological potential especially for second-generation bioethanol production (2G) from moringa straw. This is the first report on the use of M. oleifera biomass to induce the production of various cellulases in P. funiculosum. © 2019 Asociación Argentina de Microbiología. Published by Elsevier Espana, S.L.U. This is an open access article under the CC BY-NC-ND license (https://creativecommons.org/licenses/by-nc-nd/4.0/).

Resumen Actualmente, la valorización de los residuos agroindustriales es de gran interés. En este trabajo se emplearon residuos de madera blanda de Moringa oleifera para la producción de celulasas de bajo costo. El objetivo fue aislar, identificar y caracterizar microorganismos con actividad celulolítica en diferentes fuentes de carbono. A partir de la biomasa de M. oleifera, se aislaron e identificaron 42 microorganismos productores de celulasas. Los hongos que presentaron los mayores halos de hidrólisis en carboximetilcelulosa como sustrato fueron identificados molecularmente como Penicillium funiculosum (FG1), Fusarium verticillioides (FG3) y Cladosporium cladosporioides (FC2). Mediante fermentación sumergida, se evaluó la capacidad de estas cepas en la producción de celulasas utilizando celulosa cristalina (Avicel) y amorfa (CMC) como fuentes de carbono. P. funiculosum y C. cladosporioides presentaron las mayores actividades de endoglucanasa (606 U/l) y exoglucanasa (205 U/l) en medio Avicel, mientras que F. verticillioides mostró la mayor actividad de p-glucosidasa (664 U/l) en medio CMC. Además, se evaluó el efecto de tres medios de cultivo (A, B y C) sobre la producción de celulasas en P. funiculosum empleando residuos de moringa como fuente de carbono. Los resultados mostraron que en el medio C, la productividad volumétrica de celulasas se incrementó en 2,77; 8,26 y 2,30 veces para las actividades de endoglucanasa, exoglucanasa y p-glucosidasa, respectivamente. Los extractos enzimáticos producidos tienen gran potencial para su utilización biotecnológica, especialmente en la sacarificación de residuos de moringa y la producción de bioetanol de segunda generación. Este es el primer estudio del uso de la biomasa de M. oleifera para inducir la producción de diversas celulasas en P. funiculosum.

Investigation on secondary metabolites of endophytic fungus Talaromyces purpurogenus hosted in Tylophora ovate / 中国中药杂志

Eight compounds,(R)-2-[5-(methoxycarbonyl)-4-methyl-6-oxo-3,6-dihydro-2H-pyran-2-yl]acetic acid(1),(3S,4R)-3,4-dihydro-3,4-epoxy-5-hydroxynaphthalen-1(2H)-one(2),(-)-mitorubrinol(3),(-)-mitorubrin(4),(±)-asperlone A(5), terreusinone(6), verrucisidinol(7) and cerebroside C(8) were isolated from the endophytic fungus Talaromyces purpurogenus by using various column chromatographic techniques. Their structures were identified by NMR, MS, CD and optical rotation. Compounds 1 and 2 were new compounds. Their anti-diabetic activities in vitro were evaluated, and compound 1 showed moderate inhibitory activity toward XOD at 10 μmol·L~(-1) with the inhibition rate of 69.9%.

New Records of Four Species Belonging to Eurotiales from Soil and Freshwater in Korea

Four strains of Penicillium and Talaromyces species are described and illustrated in an inventory of fungal species belonging to Eurotiales. The strains, CNUFC-DDS17-1, CNUFC-DDS27-1, CNUFC-PTM72-1, and CNUFC-YJW3-31, were isolated from soil and freshwater samples from South Korea. Based on their morphological characteristics and sequence analyses by the combined β-tubulin and calmodulin gene, the CNUFC-DDS17-1, CNUFC-DDS27-1, CNUFC-PTM72-1, and CNUFC-YJW3-31 isolates were identified as Penicillium pasqualense, Penicillium sanguifluum, Talaromyces apiculatus, and Talaromyces liani, respectively. The designated strains were found to represent a previously undescribed species of Korean fungal biota. In this study, detailed morphological descriptions and phylogenetic relationships of these species are provided.

AcuD Gene Knockout Attenuates the Virulence of Talaromyces marneffei in a Zebrafish Model

Talaromyces marneffei is the only dimorphic species in its genus and causes a fatal systemic mycosis named talaromycosis. Our previous study indicated that knockdown of AcuD gene (encodes isocitrate lyase of glyoxylate bypass) of T. marneffei by RNA interference approach attenuated the virulence of T. marneffei, while the virulence of the AcuD knockout strains was not studied. In this study, T. marneffei-zebrafish infection model was successfully established through hindbrain microinjection with different amounts of T. marneffei yeast cells. After co-incubated at 28°C, the increasing T. marneffei inoculum doses result in greater larval mortality; and hyphae generation might be one virulence factor involved in T. marneffei-zebrafish infection. Moreover, the results demonstrated that the virulence of the ΔAcuD was significantly attenuated in this Zebrafish infection model.

Diversity of Aspergillus, Penicillium, and Talaromyces Species Isolated from Freshwater Environments in Korea

In order to elucidate the fungal diversity and community structure in freshwater environments, numerous fungal strains were isolated from freshwater, submerged soils, twigs, dead insects, etc. Among them, the present study has focused specifically on Aspergillus, Penicillium, and Talaromyces species, which produce diverse useful metabolites in general. Twelve strains of Aspergillus isolated were identified as A. japonicus (n = 5), A. tubingensis (3), A. niger (2), and A. flavus (2), 10 strains of which belong to Aspergillus section Nigri, named black Aspergillus. Eight strains of Penicillium were identified as P. brasilianim (n = 3), P. oxalicum (2), P. crustosum (1), P. expansum (1), and P. piscarium (1). Two different strains of Talaromyces were identified as T. pinophilus and T. versatilis. Thus far, Penicillium piscarium and Talaromyces versatilis have been unrecorded in Korea, for which we provide detailed morphological and molecular characteristics.

Characterization and structure of a novel thermostable glucoamylase from Talaromyces leycettanus JCM12802 / 生物工程学报

Glucoamylase is a critical ingredient for saccharification in the starch decomposition, and widely used in food, pharmaceutical and fermentation industries. Glucoamylases are usually thermostable and have peak activities at high temperature, as required for the industrial process of glucose production. In this study, a glucoamylase gene belonging to the glycoside hydrolase (GH) family 15, Tlga15A, was cloned from Talaromyces leycettanus JCM12802, and successfully expressed in Pichia pastoris GS115. Recombinant glucoamylase TlGA showed optimal activities at pH 4.5 and 75 °C. The result of thermostability analysis showed that TlGA retained above 70% activity after incubating for 1 h at 65 °C, and 43% residual activity after 30 min at 70 °C. Moreover, TlGA had high resistance to most metal ions and chemical reagents tested. Various starch substrates could be hydrolyzed by TlGA, including soluble starch (255.6±15.3) U/mg, amylopectin (342.3±24.7) U/mg, glycogen (185.4±12.5) U/mg, dextrin (423.3±29.3) U/mg and pullulan (65.7±8.1) U/mg. The primary, secondary and tertiary structures of glucoamylase were further analyzed. The low ratio of Gly in the primary structure and low exposed nonpolarity solvent accessible surface in the tertiary structure may be the main reasons for TlGA's thermostability. These results show that TlGA is great promising for potential use in the commercial production of glucose syrups. Moreover, this research will provide knowledge and innovating ideas for the improvement of glucoamylase thermostability.

Antimicrobial activity and acetylcholinesterase inhibition by extracts from chromatin modulated fungi

ABSTRACT Major health challenges as the increasing number of cases of infections by antibiotic multiresistant microorganisms and cases of Alzheimer's disease have led to searching new control drugs. The present study aims to verify a new way of obtaining bioactive extracts from filamentous fungi with potential antimicrobial and acetylcholinesterase inhibitory activities, using epigenetic modulation to promote the expression of genes commonly silenced. For such finality, five filamentous fungal species (Talaromyces funiculosus, Talaromyces islandicus, Talaromyces minioluteus, Talaromyces pinophilus, Penicillium janthinellum) were grown or not with DNA methyltransferases inhibitors (procainamide or hydralazine) and/or a histone deacetylase inhibitor (suberohydroxamic acid). Extracts from T. islandicus cultured or not with hydralazine inhibited Listeria monocytogenes growth in 57.66 ± 5.98% and 15.38 ± 1.99%, respectively. Increment in inhibition of acetylcholinesterase activity was observed for the extract from P. janthinellum grown with procainamide (100%), when compared to the control extract (39.62 ± 3.76%). Similarly, inhibition of acetylcholinesterase activity increased from 20.91 ± 3.90% (control) to 92.20 ± 3.72% when the tested extract was obtained from T. pinophilus under a combination of suberohydroxamic acid and procainamide. Concluding, increases in antimicrobial activity and acetylcholinesterase inhibition were observed when fungal extracts in the presence of DNA methyltransferases and/or histone deacetylase modulators were tested.

Penicillium Species Other Than Talaromyces marneffei Producing Red Pigment from Clinical Specimens: Isolation of Talaromyces albobiverticillius

No abstract available.

Primer registro de Talaromyces udagawae relacionado con cuerpos humanos en descomposición, en suelo de la Argentina / First record of Talaromyces udagawae in soil related to decomposing human remains in Argentina

The morphologic features of Talaromyces udagawae Stolk and Samson are here described and illustrated. This teleomorphic Ascomycota fungus was isolated from soil obtained in Buenos Aires province (Argentina) from beneath a human cadaver in an advanced state of decomposition. After washing and serial dilution of the soil along with moist-chamber techniques for fungal cultivation, T. udagawae formed very restricted colonies of bright yellow color on different growth media with 8-ascospored asci. The ascospores were ellipsoidal and ornamented. The anamorphic state was not observed. Molecular-genetic techniques identified the species. The present record is the first of the species in Argentina, pointing it as a tool to identify soils where cadaver decomposition occurs.

Se describen e ilustran las características morfológicas de Talaromyces udagawae Stolk y Samson. Se aisló el estado teleomórfico de este hongo Ascomycota de suelo obtenido en la provincia de Buenos Aires (Argentina), por debajo de un cadáver humano en avanzado estado de descomposición. Las muestras de suelo fueron analizadas mediante lavado, dilución seriada y cámaras húmedas, técnicas ampliamente usadas para el estudio de hongos de suelo. T. udagawae formó colonias muy restringidas de color amarillo brillante en diferentes medios de cultivo, con ascos con 8 ascosporas. Las ascosporas eran elipsoidales y ornamentadas. No fue hallado el estado anamórfico. La especie también fue identificada mediante técnicas moleculares. El presente registro es el primero de la especie en la Argentina y el único que la postula como herramienta para identificar suelos donde ocurre una descomposición cadavérica.

In vitro cytotoxicity and antimicrobial activity of Talaromyces flavus SP5 inhabited in the marine sediment of Southern Coast of India / 中国天然药物

Marine sediment samples were collected from the coastal areas of Southern India, particularly in Kanyakumari District. Twenty-eight different fungal strains were isolated. The screening of fungi from marine sediment was done to isolate a potent fungus that can produce bioactive compounds for biomedical applications. Only three strains viz Trichoderma gamsii SP4, Talaromyces flavus SP5 and Aspergillus oryzae SP6 were screened for further studies. The intracellular bioactive compounds were extracted using solvent extraction method. The crude extracts were tested for its anti-microbial and anti-cancer properties and analytically characterized using Gas Chromatography Mass Spectrometry (GC-MS). All the three extracts were active, but the extract from T. flavus SP5 was found to be more active against various human pathogens, viz., Pseudomonas aeruginosa ATCC 27853 (17.8 ± 0.1), Escherichia coli ATCC 52922 (18.3 ± 0.3), and Candida tropicalis ATCC 750 (17.7 ± 0.4). It also exhibited cytotoxic activity against HEp2 carcinoma cell line with the LC value of 25.7 μg·L. The GC-MS data revealed the presence of effective bioactive compounds. These results revealed that the extract from isolated fungus T. flavus SP5 acted as a potent antimicrobial, antifungal, and anticancer agent, providing basic information on the potency of marine fungi towards biomedical applications; further investigation may lead to the development of novel anticancer drugs.

Characterization of Paecilomyces variotii and Talaromyces amestolkiae in Korea Based on the Morphological Characteristics and Multigene Phylogenetic Analyses

During fungal diversity surveys of the order Eurotiales in Korea, two fungal strains, EML-DG33-1 and EML-NCP50, were isolated from samples of rat dung and fig tree leaf collected at a garden located in Gwangju in 2014. To complete the National Species List of Korea, it is a prerequisite to verify whether many questionable species, which were previously recorded but not confirmed, indeed present in Korea. Herein, the isolates were confirmed as undescribed species, Paecilomyces variotii and Talaromyces amestolkiae based on the combination of morphological and phylogenetic analyses of multigenes including the rDNA internal transcribed spacer, β-tubulin, and RNA polymerase II subunit 2.

Comparison of Talaromyces marneffei Infection in Human Immunodeficiency Virus-positive and Human Immunodeficiency Virus-negative Patients from Fujian, China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Talaromyces (Penicillium) marneffei (TM) is an emerging dimorphic human pathogenic fungus that is endemic to Southeast Asia. TM mostly occurs as an opportunistic infection in patients with human immunodeficiency virus (HIV). The objective of this study was to compare the clinical and laboratory parameters of patients with TM infections who were HIV-positive and HIV-negative and to assess therapies and outcomes.</p><p><b>METHODS</b>This was a retrospective analysis of 26 patients diagnosed with disseminated TM infection from September 2005 to April 2014 at Fujian Provincial Hospital, China.</p><p><b>RESULTS</b>Patients with TM infection tend to present with fever, weight loss, and anemia. The time from symptom onset to confirmed diagnosis was greater for HIV-negative patients (n = 7; median: 60 days, range: 14-365 days) than for HIV-positive patients (n = 19; median: 30 days, range: 3-90 days, Mann-Whitney U = 31.50, P= 0.041). HIV-negative patients were more likely to have dyspnea (57.1% vs. 5.3%, χ2 = 8.86, P= 0.010), low neutrophil count (Mann-Whitney U = 27.00, P= 0.029), high CD4 count (Mann-Whitney U = 0.00, P= 0.009), and high lymphocyte count (Mann-Whitney U = 21.00, P= 0.009). There were no significant differences in other demographic, clinical, or biochemical characteristics. Among all the patients, 12 HIV-positive patient and 1 HIV-negative patient received amphotericin and fluconazole treatment, 9 of whom improved, 1 died, 2 had kidney damage, 1 had hypokalemia due to exceeded doses.</p><p><b>CONCLUSIONS</b>HIV-negative patients with TM infections tend to have a longer diagnostic interval, a higher percentage of dyspnea, higher levels of CD4 and lymphocytes, and lower neutrophil counts than TM infection in HIV-positive patients. Treatment programs with amphotericin and fluconazole are mostly effective.</p>

Diversity of Endophytic Fungi Associated with the Roots of Four Aquatic Plants Inhabiting Two Wetlands in Korea

A total of 4 aquatic plants, Eleocharis kuroguwai Ohwi, Hydrocharis dubia Backer, Salvinia natans All., and Zizania latifolia Turcz., were sampled from representative two wetlands of South Korea. A total of 38 endophytic fungal strains were isolated from aquatic plants native to the Daepyeong wetland, and 27 strains were isolated from the Jilnal wetland. The internal transcribed spacer regions of fungal isolates were sequenced and a phylogenetic analysis was performed. In addition, endophytic fungal diversity from each wetland and host plant species was deduced. A total of 25 fungal genera were purely isolated, and 16 fungal genera were isolated from each of the two wetlands. Commonly isolated genera from both wetlands were Aspergillus, Cladosporium, Clonostachys, Fusarium, Leptosphaeria, Penicillium, and Talaromyces. This study revealed that fungal diversity varied with environmental conditions and by host plant in representative two wetlands.

Diversity and Plant Growth Promoting Capacity of Endophytic Fungi Associated with Halophytic Plants from the West Coast of Korea

Five halophytic plant species, Suaeda maritima, Limonium tetragonum, Suaeda australis, Phragmites australis, and Suaeda glauca Bunge, which are native to the Muan salt marsh of South Korea, were examined for fungal endophytes by sequencing the internal transcribed spacer (ITS) region containing ITS1, 5.8S rRNA, and ITS2. In total, 160 endophytic fungal strains were isolated and identified from the roots of the 5 plant species. Taxonomically, all 160 strains belonged to the phyla Ascomycota, Basidiomycota, and Zygomycota. The most dominant genus was Fusarium, followed by the genera Penicillium and Alternaria. Subsequently, using 5 statistical methods, the diversity indices of the endophytes were determined at genus level. Among these halophytic plants, P. australis was found to host the greatest diversity of endophytic fungi. Culture filtrates of endophytic fungi were treated to Waito-C rice seedlings for plant growth-promoting effects. The fungal strain Su-3-4-3 isolated from S. glauca Bunge provide the maximum plant length (20.1 cm) in comparison with wild-type Gibberella fujikuroi (19.6 cm). Consequently, chromatographic analysis of the culture filtrate of Su-3-4-3 showed the presence of physiologically active gibberellins, GA(1) (0.465 ng/mL), GA(3) (1.808 ng/mL) along with other physiologically inactive GA(9) (0.054 ng/mL) and GA(24) (0.044 ng/mL). The fungal isolate Su-3-4-3 was identified as Talaromyces pinophilus.

Discovery of Two New Talaromyces Species from Crop Field Soil in Korea

Two new fungal species of the genus Talaromyces, Talaromyces purpurogenus and Talaromyces trachyspermus from the Trichocomaceae family, were recovered during an investigation of fungal communities in soil collected from the Gangwon-do and Jeollanam-do provinces of Korea. These two species have not been previously officially reported from Korea. In this study, detailed descriptions of internal transcribed spacer rDNA and beta-tubulin gene regions of these two fungi are presented. Morphological features of the two fungi in five agar media, potato dextrose, oatmeal, malt extract, czapek yeast extract, and yeast extract sucrose, are also reported. The species were identified on the basis of molecular and morphological analysis, and herein we present data with detailed descriptions and figures.

Two new polyesters from wetland soil-derived fungus Talaromyces flavus / 中国中药杂志

Two new polyesters, talapolyesters G-H (1-2) were isolated from the wetland soil-derived fungus Talaromyces flavus BYD07-13, and their structures were determined by NMR and MS spectroscopic data. The absolute configurations of the residues were determined by alkaline hydrolysis. The cytotoxicity against five tumor cell lines (HL-60, SMMC-7721, A-549, MCF-7 and SW480) of 1-2 was examined.

Diversity of Culturable Soil Micro-fungi along Altitudinal Gradients of Eastern Himalayas

Very few studies have addressed the phylogenetic diversity of fungi from Northeast India under the Eastern Himalayan range. In the present study, an attempt has been made to study the phylogenetic diversity of culturable soil fungi along the altitudinal gradients of eastern Himalayas. Soil samples from 24 m above sea level to 2,000 m above sea level altitudes of North-East India were collected to investigate soil micro-fungal community structure and diversity. Molecular characterization of the isolates was done by PCR amplification of 18S rDNA using universal primers. Phylogenetic analysis using BLAST revealed variation in the distribution and richness of different fungal biodiversity over a wide range of altitudes. A total of 107 isolates were characterized belonging to the phyla Ascomycota and Zygomycota, corresponding to seven orders (Eurotiales, Hypocreales, Calosphaeriales, Capnodiales, Pleosporales, Mucorales, and Mortierellales) and Incertae sedis. The characterized isolates were analysed for richness, evenness and diversity indices. Fungal diversity had significant correlation with soil physico-chemical parameters and the altitude. Eurotiales and Hypocreales were most diverse and abundant group of fungi along the entire altitudinal stretch. Species of Penicillium (D = 1.44) and Aspergillus (D = 1.288) were found to have highest diversity index followed by Talaromyces (D = 1.26) and Fusarium (D = 1.26). Fungal distribution showed negative correlation with altitude and soil moisture content. Soil temperature, pH, humidity and ambient temperature showed positive correlation with fungal distribution.