Antifungal activity of marine-derived actinomycetes against Talaromyces marneffei.

AIMS: This study aimed to isolate actinomycetes from marine environments and examine their antifungal activity against Talaromyces marneffei both in vitro and in vivo. METHODS AND RESULTS: Nineteen out of 101 actinomycete extracts were active and further determined for their minimum inhibitory concentrations (MIC). Three extracts of AMA50 that isolated from sediment showed strong antifungal activity against T. marneffei yeast (MICs ≤0·03-0·25 µg ml-1 ) and mould (MICs 0·5-16 µg ml-1 ) forms. The hexane extract from the cells of AMA50 (AMA50CH) exhibited the best activity against both the forms (MIC ≤ 1 µg ml-1 ). Three extracts from AMA50 killed the melanized yeast cells at 0·5 µg ml-1 . The AMA50CH was further tested for protective effects in Caenorhabditis elegans model. At concentrations of 1-8 µg ml-1 , the AMA50CH prolonged survival of T. marneffei-infected C. elegans with a 60-70% survival rate. The composition of AMA50CH was determined by gas chromatography-mass spectrometry. The major components were n-hexadecanoic acid, tetradecanoic acid and pentadecanoic acid. Sequencing analysis revealed that isolate AMA50 belonged to the genus Streptomyces. CONCLUSIONS: The AMA50CH from Streptomyces sp. AMA50 was the most effective extract against T. marneffei. SIGNIFICANCE AND IMPACT OF THE STUDY: Talaromyces marneffei is one of the most important thermally dimorphic pathogenic fungi. These results indicated the potency of marine-derived actinomycete extracts against T. marneffei both in vitro and in vivo.

Comparative studies on Pb(II) biosorption with three spongy microbe-based biosorbents: High performance, selectivity and application.

Lead pollution in industrial-derived water has become an increasingly serious concern. The development of adsorbents with excellent efficiency, selectivity and separability using diverse microorganisms is ideal for treating lead pollution. In this study, gram-negative bacteria Pseudomonas putida I3, gram-positive bacteria Microbacterium sp. OLJ1 and mycelial fungus Talaromyces amestolkiae Pb served as raw materials to facilely synthesize sponge-like biosorbents via a one-step method at room temperature. SEM, EDS, FTIR, 13C NMR, XRD and XPS were used for investigating the morphology and surface properties of these three biosorbents. The obtained biosorbents possessed the same three-dimensional porous structure but different productivities and mechanical strengths due to the similar chemical compositions and different cell structures of their microorganisms. Pb(II) adsorption on X-PI3, X-OLJ1 and X-TPb was fast and pH dependent, with maximal adsorption capacities of 345.02, 237.02 and 199.02 mg/g, respectively. The biosorbents had a high selectivity for Pb(II), while Pb(II) remarkably suppressed the adsorption of co-existing heavy metal ions. The analyses indicated that Pb(II) removal was mainly achieved by ion exchange reactions, surface complexation with heteroatom-containing functional groups and microprecipitation. The treatment effects of synthetic and real wastewater revealed that the as-prepared biosorbents are promising for Pb(II) removal.

Discovery of a sexual cycle in Talaromyces amestolkiae.

Talaromyces amestolkiae is a common cosmopolitan species that has been cultured from indoor house dust, sputum and lungs from cystic fibrosis patients, indoor air, wheat, soil, pineapple, sculptures and manure. It was described as an asexual Talaromyces species and was reported to produce black sclerotia. In this study we report on the induction of sexual reproductive structures in T. amestolkiae. The mating type of 18 T. amestolkiae strains was determined with MAT-specific primers. Subsequently opposite mating types were inoculated on oatmeal agar and malt-extract agar and incubated 6-20 wk at 25 and 30 C in darkness. After incubation single ascospore isolations were made and evidence of recombination in the offspring was examined by amplified fragment length polymorphism and pairwise homoplasy index test, which is implemented in Splitstree4. The offspring displayed clear evidence of recombination on a genetic level as shown in the variations observed between banding patterns in the amplified fragment length polymorphism. Also a net-like and reticulated NeighborNet was observed and the pairwise homoplasy index test for recombination supported the presence of recombination (P = 0.003372). The distribution of MAT1-1 and MAT1-2 genes in the progeny showed a close to 1:1 ratio. Talaromyces amestolkiae is only the second heterothallic Talaromyces species to produce ascomata and ascospores under laboratory conditions.

Heat activation of Neosartorya and Talaromyces ascospores and enhancement by organic acids.

Neosartorya and Talaromyces are typical fungi capable of producing heat resistant ascospores responsible for the spoilage of processed fruit products. In this study, the heat activation rates of Neosartorya and Talaromyces ascospores were investigated in several suspending media at various heating temperatures. Ascospores were dispersed in pH 3.5 McIlvain buffer, organic acid/alcohol-supplemented McIlvain buffer and grape juice (pH 3.5, 5.0 degrees Brix) prior to heat treatments. In McIlvain buffer, the number of germinating ascospores increased logarithmically with longer exposure to heating at an test temperatures. Heat activation rates (k values) accelerated with increasing temperature. The calculated activation energy (Ea) values were similar among ascospores from the same genus, but the Ea of the test Neosartorya spp. were greater than that of the test Talaromyces spp. Greater k values were calculated from acetate-supplemented McIlvain buffer and grape juice. Similarly, normal- and branched-chain fatty acids were shown to enhance the heat activation rate of the ascospores in McIlvain buffer systems. These results could assist the food industry in designing adequate thermal processes for food products against the heat resistant fungi.

High viscosity and anisotropy characterize the cytoplasm of fungal dormant stress-resistant spores.

Ascospores of the fungus Talaromyces macrosporus are dormant and extremely stress resistant, whereas fungal conidia--the main airborne vehicles of distribution--are not. Here, physical parameters of the cytoplasm of these types of spores were compared. Cytoplasmic viscosity and level of anisotropy as judged by spin probe studies (electron spin resonance) were extremely high in dormant ascospores and during early germination and decreased only partly after trehalose degradation and glucose efflux. Upon prosilition (ejection of the spore), these parameters fell sharply to values characteristic of vegetative cells. These changes occurred without major volume changes that suggest dramatic changes in cytoplasmic organization. Azide reversibly inhibited prosilition as well as the decline in cytoplasmic parameters. No organelle structures were observed in etched, cryoplaned specimens of ascospores by low-temperature scanning electron microscopy (LTSEM), confirming the high cytoplasmic viscosity. However, cell structures became visible upon prosilition, indicating reduced viscosity. The viscosity of fresh conidia of different Penicillium species was lower, namely, 3.5 to 4.8 cP, than that of ascospores, near 15 cP. In addition the level of anisotropic motion was markedly lower in these cells (h(0)/h(+1) = 1.16 versus 1.4). This was confirmed by LTSEM images showing cell structures. The decline of cytoplasmic viscosity in conidia during germination was linked with a gradual increase in cell volume. These data show that mechanisms of cytoplasm conservation during germination differ markedly between ascospores and conidia.

Dormant ascospores of Talaromyces macrosporus are activated to germinate after treatment with ultra high pressure.

AIMS: Ascospores of Talaromyces macrosporus are constitutively dormant and germinate after a strong external shock, classically a heat treatment. This fungus is used as a model system to study heat resistance leading to food spoilage after pasteurization. This study evaluates the effect of high pressure on the germination behaviour of these spores. METHODS AND RESULTS: Ascospore containing bags were subjected to ultra high pressure and spores were plated out on agar surfaces. Untreated suspensions showed invariably very low germination. Increased germination of ascospores occurred after short treatments at very high pressure (between 400 and 800 MPa). Activation is partial compared with heat activation and did not exceed 6.9% (65 times that of untreated suspensions) of the spore population. Maximum activation was attained shortly (10 s-3 min) after the pressure was applied and accompanied by cell wall deformations as judged by scanning electron microscopy. The spores observed in this study were harvested from cultures that were 39-58 days old. The maturity of spores at similar developmental stages was measured by assessing the heat resistance of ascospores. Between 20 and 40 days heat resistance increased 2.4-fold, but only an additional increase of 1.3-fold was observed at later stages (40-67 days). CONCLUSIONS: Our investigations show that high pressure constitutes a second type of shock that can activate heat-resistant ascospores to germinate. Activation is maximal after very short treatments and accompanied with changes in the cell wall structure. High-pressure activation is not the result of immaturity of the ascospores. SIGNIFICANCE AND IMPACT OF THE STUDY: These observations are relevant for the application of high pressure as a novel pasteurization method.