Mycology-Nanotechnology Interface: Applications in Medicine and Cosmetology.

In today's time, nanotechnology is being utilized to develop efficient products in the cosmetic and pharmaceutical industries. The application of nanotechnology in transforming bioactive material into nanoscale products substantially improves their biocompatibility and enhances their effectiveness, even when used in lower quantities. There is a significant global market potential for these nanoparticles because of which research teams around the world are interested in the advancements in nanotechnology. These recent advances have shown that fungi can synthesize metallic nanoparticles via extra- and intracellular mechanisms. Moreover, the chemical and physical properties of novel metallic nanoparticles synthesised by fungi are improved by regulating the surface chemistry, size, and surface morphology of the nanoparticles. Compared to chemical synthesis, the green synthesis of nanoparticles offers a safe and sustainable approach for developing nanoparticles. Biosynthesised nanoparticles can potentially enhance the bioactivities of different cellular fractions, such as plant extracts, fungal extracts, and metabolites. The nanoparticles synthesised by fungi offer a wide range of applications. Recently, the biosynthesis of nanoparticles using fungi has become popular, and various ways are being explored to maximize nanoparticles synthesis. This manuscript reviews the characteristics and applications of the nanoparticles synthesised using the different taxa of fungi. The key focus is given to the applications of these nanoparticles in medicine and cosmetology.

Identification of antibiotics for use in selection of the chytrid fungi Batrachochytrium dendrobatidis and Batrachochytrium salamandrivorans.

Global amphibian populations are being decimated by chytridiomycosis, a deadly skin infection caused by the fungal pathogens Batrachochytrium dendrobatidis (Bd) and B. salamandrivorans (Bsal). Although ongoing efforts are attempting to limit the spread of these infections, targeted treatments are necessary to manage the disease. Currently, no tools for genetic manipulation are available to identify and test specific drug targets in these fungi. To facilitate the development of genetic tools in Bd and Bsal, we have tested five commonly used antibiotics with available resistance genes: Hygromycin, Blasticidin, Puromycin, Zeocin, and Neomycin. We have identified effective concentrations of each for selection in both liquid culture and on solid media. These concentrations are within the range of concentrations used for selecting genetically modified cells from a variety of other eukaryotic species.

Folk classification of wild mushrooms from San Isidro Buensuceso, Tlaxcala, Central Mexico.

BACKGROUND: An ethnomycological study was conducted to describe the fungus concept and the traditional fungus classification system for the Nahuas of San Isidro Buensuceso, in central Mexico. The study which provides information on the co-existence of various forms of classification, based on both cultural and biological characteristics. METHODS: The research included conducting community interviews and forest forays in the company of mushroom pickers. The triad technique, pile sorting, and fresh mushroom sampling methods were used. Traditional names were analyzed to describe the Nahua classification system for fungi. RESULTS AND CONCLUSION: The triad technique with non-utilitarian stimuli allowed the fungi to be identified as an independent group of plants and animals. The Nahua people of San Isidro classify fungi primarily based on their use, where they grow, and by humoral characteristics. The analysis of the names revealed a classification based on the criteria proposed by Brent Berlin. This study identified the detailed knowledge of fungi in this Nahua community. The criteria used for the recognition of the species are very reliable, since they use organoleptic, ecological, phenological, and morphological characteristics.

3-Methylxanthine production through biodegradation of theobromine by Aspergillus sydowii PT-2.

BACKGROUND: Methylxanthines, including caffeine, theobromine and theophylline, are natural and synthetic compounds in tea, which could be metabolized by certain kinds of bacteria and fungi. Previous studies confirmed that several microbial isolates from Pu-erh tea could degrade and convert caffeine and theophylline. We speculated that these candidate isolates also could degrade and convert theobromine through N-demethylation and oxidation. In this study, seven tea-derived fungal strains were inoculated into various theobromine agar medias and theobromine liquid mediums to assess their capacity in theobromine utilization. Related metabolites with theobromine degradation were detected by using HPLC in the liquid culture to investigate their potential application in the production of 3-methylxanthine. RESULTS: Based on theobromine utilization capacity, Aspergillus niger PT-1, Aspergillus sydowii PT-2, Aspergillus ustus PT-6 and Aspergillus tamarii PT-7 have demonstrated the potential for theobromine biodegradation. Particularly, A. sydowii PT-2 and A. tamarii PT-7 could degrade theobromine significantly (p < 0.05) in all given liquid mediums. 3,7-Dimethyluric acid, 3-methylxanthine, 7-methylxanthine, 3-methyluric acid, xanthine, and uric acid were detected in A. sydowii PT-2 and A. tamarii PT-7 culture, respectively, which confirmed the existence of N-demethylation and oxidation in theobromine catabolism. 3-Methylxanthine was common and main demethylated metabolite of theobromine in the liquid culture. 3-Methylxanthine in A. sydowii PT-2 culture showed a linear relation with initial theobromine concentrations that 177.12 ± 14.06 mg/L 3-methylxanthine was accumulated in TLM-S with 300 mg/L theobromine. Additionally, pH at 5 and metal ion of Fe2+ promoted 3-methylxanthine production significantly (p < 0.05). CONCLUSIONS: This study is the first to confirm that A. sydowii PT-2 and A. tamarii PT-7 degrade theobromine through N-demethylation and oxidation, respectively. A. sydowii PT-2 showed the potential application in 3-methylxanthine production with theobromine as feedstock through the N-demethylation at N-7 position.

Antifungal susceptibility testing practices in mycology laboratories in France, 2018.

A survey of mycology laboratories for antifungal susceptibility testing (AFST) was undertaken in France in 2018, to better understand the difference in practices between the participating centers and to identify the difficulties they may encounter as well as eventual gaps with published standards and guidelines. The survey captured information from 45 mycology laboratories in France on how they perform AFST (number of strains tested, preferred method, technical and quality aspects, interpretation of the MIC values, reading and interpretation difficulties). Results indicated that 86% of respondents used Etest as AFST method, with a combination of one to seven antifungal agents tested. Most of the participating laboratories used similar technical parameters to perform their AFST method and a large majority used, as recommended, internal and external quality assessments. Almost all the participating mycology laboratories (98%) reported difficulties to interpret the MIC values, especially when no clinical breakpoints are available. The survey highlighted that the current AFST practices in France need homogenization, particularly for MIC reading and interpretation.

Professor Shu-Ting Chang on His 90th Birthday.

This issue is dedicated to the 90th birthday of Professor Shu-Ting Chang, a prominent scientist in the field of mushroom biology, including cultivation and nutritional values of mushrooms, medicinal mushroom science, and environmental impact of mushrooms. Professor Shu-Ting Chang is also one of the initiators and has been an editor of our journal from its inception (in 1999) until now.

Ethnomycological knowledge among Kaqchikel, indigenous Maya people of Guatemalan Highlands.

BACKGROUND: The Guatemalan Highlands is a region of great but so far poorly known mycological diversity. People living in this area have long used wild fungi as a source of food and income. However, our knowledge of the ethnomycological practices of the Mayan peoples of Guatemala is still rudimental, especially if compared with information reported for the neighboring region of Mexico. Among the main indigenous groups of the Maya people inhabiting the highlands of Central Guatemala, stand the Kaqchikel, accounting for nearly 8% of the entire Guatemalan population. The main aim of this study was to record the traditional knowledge and use of edible wild mushrooms by inhabitants of the municipality of San Juan Sacatepéquez that lies at the heart of the Kaqchikel area in the central highlands of Guatemala, also describing the relevant selling practices and dynamics. A secondary aim was to compare the diversity and composition of the mushroom assemblage offered at the market with the macrofungal diversity of woods in the area. METHODOLOGY: This study is the result of 4 years of ethnomycological research, conducted through continuous visits to the municipal market and focused interviews with collectors and vendors. Field sampling in pine-oak forested areas surrounding San Juan Sacatepéquez, from where the mushrooms sold at the market are foraged, were also conducted, in the presence of local collectors. RESULTS: The results show a significant richness of species sold in the market, a network of commerce of purchase, sale, and resale of several species, with relatively stable prices, and knowledge about edible and inedible species that is transmitted mainly within the family nucleus. The business of selling mushrooms in the market is an exclusive activity of women, who are supplied by collectors or by other vendors. Fungi are sold and bought only as food, while no consumption of hallucinogenic mushrooms or medicinal mushrooms was recorded. Several species of Amanita, Cantharellus, Boletus, Lactarius, and Russula were those most commercialized in the 4 years of the study, but we also spotted fungi never reported before as consumed in the country, including Gastropila aff. fumosa (= Calvatia fumosa) and several species of Cortinarius. Field sampling in nearby pine-oak forests confirmed an elevated local macrofungal diversity. CONCLUSION: Our study unveiled the contemporary wealth of Kaqchikel culture for what concerns mushrooms, demonstrating that mushrooms continue to be culturally and economically important for these communities despite the erosion of traditional knowledge. Our results also confirmed the need to investigate in greater detail the Guatemalan mycodiversity that is vast and poorly known.

Metabolic profiling of natural and cultured Cordyceps by NMR spectroscopy.

Cordyceps, a type of Chinese herbal medicine that exhibits anti-angiogenesis and tumor growth suppression effects, has recently gained increasing popularity. However, high-quality, natural Cordyceps, such as Ophiocordyceps sinensis, is very rare and difficult to obtain in large amounts. Cordyceps is cultured instead of harvested from natural sources, but the quality with respect to the ingredients has not been fully studied. In this study, we performed an NMR metabolic profiling of aqueous extracts of Cordyceps without any sample treatment to evaluate the proper species and medium and influence of two different disinfection methods. It was discovered that Cordyceps militaris fungus and silkworm chrysalis medium were suitable for cultivation of Cordyceps. Furthermore, cordycepin, a Cordyceps-specific functional compound, was produced at different growth stages during different cultivation processes, even at the mycelial stage, and was found at three times higher concentrations in cultured C. militaris compared to that in naturally occurring C. militaris.

Ethnomycological knowledge in three communities in Amealco, Quéretaro, México.

BACKGROUND: Fungi have multiple uses in temperate areas of México, but an important decrease in the traditional knowledge of uses and customs of mushrooms becomes a fundamental issue for fungi conservation. However, only few studies quantify the traditional ethnomycological knowledge in México, and this study is the first quantitative report for Querétaro, a central state with both Otomí and Mestizo communities and a high fungi diversity. METHODS: The present study was conducted registering traditional knowledge on the use and consumption of mushrooms in three Hñähñu (Otomí) communities (Tesquedó, Xajay, and Tenasdá) in Amealco de Bonfil, Querétaro, México, between August 2013 and November 2014. We conducted a stratified sampling, where uses common Hñähñu and Spanish names, and eight quantitative variables that conform the "Edible Mushrooms Cultural Significant Index" (EMCI) were recorded from 100 informants. For the classification and ordination analysis of species and uses, we used multivariate techniques such as cluster, multidimensional scaling, and principal components (PC). RESULTS: Thirty-three mushrooms species were registered, most of them used for consumption by households, few aimed for commercial purposes, one species is medicinal, another has veterinary, and other ludic uses (as a toy). The three species with the highest EMCSI were Amanita basii, Fistulinella wolfeana, and Lactarius indigo. Edibility was the main use detected in the survey, and people harvested mushrooms provided by the forest mainly during the rainy season. We observed that mushroom searching and collection are activities that strengthen the family ties and are crucial for the transfer of this knowledge through generations. Cluster analysis separates groups according to different values in EMCSI variables, and principal components ordinate the species by frequencies (PC1) and traditions (PC2). CONCLUSIONS: The current state of knowledge in the studied communities is strong, especially among women, but with a tendency to disappear due to migration and lack of interest among new generations. Future quantitative studies are important to analyze tendencies of the traditional ethnomycological knowledge transferred to new generations.

Cultivation of Agaricus bisporus enriched with selenium, zinc and copper.

BACKGROUND: Agaricus bisporus (white button mushroom) is an important culinary and medicinal species of worldwide importance. The present study investigated for the first time whether it may be grown on substrates supplemented with Se alone or in combination with Cu and/or Zn (0.1-0.8 mmol L-1 ) to produce fruiting bodies of increased nutritional value. RESULTS: As found, substrate supplementation did not affect yielded biomass up to 0.6 mmol L-1 element concentrations regardless of the cultivation model. At 0.8 mmol L-1 Se + Cu and Se + Zn supplementation biomass comparable with controls still developed. The accumulation of trace elements in the fruiting bodies generally increased over the concentration gradient reaching its maximum at 0.6 mmol L-1 (for Se + Zn and Se + Cu + Zn) and 0.8 mmol L-1 (for Se and Se + Cu). The organic Se constituted the greatest share in total Se quota. As calculated, each 10 g of dried fruiting bodies of A. bisporus obtained from 0.6 or 0.8 mmol L-1 supplementation would represent 342-469% of the Recommended Daily Allowance (RDA) for Se, 43.4-48.5% for Cu and 5.2-5.8% for Zn. CONCLUSION: Considering inexpensive methods of A. bisporus cultivation, global popularity and use of this mushroom, its biofortification with Se, Cu and Zn could have a practical application in deficiency prevention and assisted treatment. © 2016 Society of Chemical Industry.

Rarely reported fungal spores and structures: An overlooked source of probative trace evidence in criminal investigations.

The value of pollen and plant spores as trace evidence has long been established, but it is only in the last eight years that fungal spores have been analysed routinely from the same palynological samples. They have greatly enhanced the specificity of links between people, objects, and places. Most fungal species occupy restricted ecological niches and their distributions can be limited both spatially and geographically. Spores may be dispersed over very short distances from the fungal sporophore,(1) and their presence in any palynological assemblage may indicate a restricted area of ground, or the presence of particular plants (even specific dead plant material). Fungal spores can represent primary, secondary, or even tertiary proxy evidence of a location, and can indicate the presence of a plant even though the plant is not obvious at a crime scene. In some cases, spores from fungi which have rarely been reported, and are considered to be rare, have been of particular value in providing intelligence or evidence of contact. Ten examples are given from case work in which rarely reported or unusual fungi have proved to be important in criminal investigations.

Problemas clínicos en micología médica: problema número 49 / Clinical problems in medical mycology: problem number 49

The case of a 59-year-old female born in Buenos Aires (Argentina) is presented. She had been diagnosed with HIV in 2007 and received highly active antiretroviral therapy until 2011; she also suffered from diabetes type 2. She had received empirical treatment (pyrimethamine-clindamycin) for cerebral toxoplasmosis. Fifteen days later she suffered a drug-induced skin disorder and was treated in the Dermatology Service of the Hospital Muñiz with corticosteroids. After five weeks she was readmitted to the Infectious Disease Unit due to asthenia, weight loss, left hip pain and weakness in all four limbs. Septic arthritis and aseptic hip necrosis were ruled out. Blood cultures were positive for Staphylococcus aureus and Escherichia coli. The patient received intravenous antibiotics, but before being discharged Acinetobacter baumannii was isolated from blood, catheter and urine cultures, and a new series of antibiotics were prescribed. On the 3rd day she presented encephalic facies, changes of behaviour and disorientation, without nuchal rigidity, Kernig and Brudzinski signs or focal signs. An X-ray computed tomography did not show parenchymal lesions. A yeast identified as Candida albicans was isolated in a cerebrospinal fluid culture. The same yeast was recovered in a new cerebrospinal fluid sample. The isolate was susceptible to amphotericin B and susceptible dose dependent to fluconazole. The patient was treated with amphotericin B (0.7 mg/kg plus 800 mg fluconazole daily). Three weeks later, new cerebrospinal fluid cultures were negative. Unfortunately, the patient died soon afterwards (AU)

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Nutrient control for stationary phase cellulase production in Trichoderma reesei Rut C-30.

This work describes the use of nutrient limitations with Trichoderma reesei Rut C-30 to obtain a prolonged stationary phase cellulase production. This period of non-growth may allow for dependable cellulase production, extended fermentation periods, and the possibility to use pellet morphology for easy product separation. Phosphorus limitation was successful in halting growth and had a corresponding specific cellulase production of 5±2 FPU/g-h. Combined with the addition of Triton X-100 for fungal pellet formation and low shear conditions, a stationary phase cellulase production period in excess of 300 h was achieved, with a constant enzyme production rate of 7±1 FPU/g-h. While nitrogen limitation was also effective as a growth limiter, it, however, also prevented cellulase production.

Miniaturized Cultivation of Microbiota for Antimalarial Drug Discovery.

The ongoing search for effective antiplasmodial agents remains essential in the fight against malaria worldwide. Emerging parasitic drug resistance places an urgent need to explore chemotherapies with novel structures and mechanisms of action. Natural products have historically provided effective antimalarial drug scaffolds. In an effort to search nature's chemical potential for antiplasmodial agents, unconventionally sourced organisms coupled with innovative cultivation techniques were utilized. Approximately 60,000 niche microbes from various habitats (slow-growing terrestrial fungi, Antarctic microbes, and mangrove endophytes) were cultivated on a small-scale, extracted, and used in high-throughput screening to determine antimalarial activity. About 1% of crude extracts were considered active and 6% partially active (≥ 67% inhibition at 5 and 50 µg/mL, respectively). Active extracts (685) were cultivated on a large-scale, fractionated, and screened for both antimalarial activity and cytotoxicity. High interest fractions (397) with an IC50 < 1.11 µg/mL were identified and subjected to chromatographic separation for compound characterization and dereplication. Identifying active compounds with nanomolar antimalarial activity coupled with a selectivity index tenfold higher was accomplished with two of the 52 compounds isolated. This microscale, high-throughput screening project for antiplasmodial agents is discussed in the context of current natural product drug discovery efforts.

Adjustment of a rapid method for quantification of Fusarium spp. spore suspensions in plant pathology.

The use of a Neubauer chamber is a broadly employed method when cell suspensions need to be quantified. However, this technique may take a long time and needs trained personnel. Spectrophotometry has proved to be a rapid, simple and accurate method to estimate the concentration of spore suspensions of isolates of the genus Fusarium. In this work we present a linear formula to relate absorbance measurements at 530nm with the number of microconidia/ml in a suspension.

Effect of revulsive cultivation on the yield and quality of newly formed sclerotia in medicinal Wolfiporia cocos.

Wolfiporia cocos is a well-known medicinal mushroom widely used in China, Japan and other Asiatic countries for its various therapeutic effects. 'Revulsive cultivation' is a newly developed method for promoting sclerotia growth in W. cocos field cultivation in China. In this report, we have systematically examined the effects of 'revulsive cultivation' on the yield and quality of newly formed sclerotia. The results showed that the genetic differences between the cultivated strain and the revulsive strain of T1 used in this study did not affect the formation process of new, large sclerotia in which the mycelia of the cultivated strain grew on pine logs directionally assembled on the revulsive strain. Additionally, 'revulsive cultivation', in which the cultivated strain and the revulsive strain used had the same or different genotypes, could remarkably increase the yield, lower the water content, and increase the water-soluble polysaccharide content of the newly formed sclerotia. Moreover, we observed that the changes in the values of the tested economic traits obtained from different genotype combinations through 'revulsive cultivation' were dissimilar. The correlations of these changes with the original sclerotium-forming ability of the cultivated strains and the genetic differences between the cultivated strain and the revulsive strain were not significant. These results will broaden our knowledge regarding the field cultivation of this medical fungus, stimulate new thinking on the study of sclerotium formation in some sclerotium-forming fungi, and promote further studies on the mechanism of sclerotium formation in W. cocos.

Comparative evaluation of antifungal activity of melaleuca oil and fluconazole when incorporated in tissue conditioner: an in vitro study.

PURPOSE: This in vitro study sought to compare the antifungal activity of melaleuca alternifolia oil and fluconazole mixed with a tissue conditioner. MATERIALS AND METHODS: By testing several concentrations of fluconazole and melaleuca oil in Visco-gel, the minimum most effective concentration of each antifungal agent against Candida albicans was determined. Mean inhibition diameter (MID) was used to measure the antifungal activity, and data were analyzed statistically for significance of findings. To determine the minimum most effective concentration of fluconazole, different concentrations of 1%, 3%, 5%, and 10% w/w in Visco-gel were tested on Sabouraud dextrose agar pregrown with C. albicans. MIDs were measured at 24 hours and on day 7, while carrying out the monitoring every day. Similarly, the minimum most effective concentration of melaleuca oil in Visco-gel was found by testing it in several concentrations (1%, 5%, 10%, 20%, 25%, 27.5%, 30%, 35% w/w). Subsequently, the minimum most effective concentration of each antifungal agent was used to compare the antifungal activity against C. albicans over 7 days using the same procedure and using plain tissue conditioner as the control. RESULT: The minimum most effective concentrations of melaleuca oil in Visco-gel and fluconazole in Visco-gel were 30% w/w and 5% w/w, respectively. Thirty percent w/w melaleuca oil was found to be the most effective (p < 0.001) and superior to 5% fluconazole in Visco-gel, as it retained substantial antifungal activity (MID), even on day 7 when fluconazole had lost its antifungal effect completely as evidenced by regrowth of C. albicans by day 7. CONCLUSION: Thirty percent melaleuca oil in tissue-conditioner Visco-gel was superior to 5% fluconazole in Visco-gel as an antifungal agent. Though both showed comparable antifungal activity at 24 hours against C. albicans, fluconazole had completely lost it by day 7, whereas melaleuca oil had substantially retained its antifungal action.