Expert Mycologists Research Articles

Diagnosis of Soybean Diseases Caused by Fungal and Oomycete Pathogens: Existing Methods and New Developments

Soybean (Glycine max) acreage is increasing dramatically, together with the use of soybean as a source of vegetable protein and oil. However, soybean production is affected by several diseases, especially diseases caused by fungal seed-borne pathogens. As infected seeds often appear symptomless, diagnosis by applying accurate detection techniques is essential to prevent propagation of pathogens. Seed incubation on culture media is the traditional method to detect such pathogens. This method is simple, but fungi have to develop axenically and expert mycologists are required for species identification. Even experts may not be able to provide reliable type level identification because of close similarities between species. Other pathogens are soil-borne. Here, traditional methods for detection and identification pose even greater problems. Recently, molecular methods, based on analyzing DNA, have been developed for sensitive and specific identification. Here, we provide an overview of available molecular assays to identify species of the genera Diaporthe, Sclerotinia, Colletotrichum, Fusarium, Cercospora, Septoria, Macrophomina, Phialophora, Rhizoctonia, Phakopsora, Phytophthora, and Pythium, causing soybean diseases. We also describe the basic steps in establishing PCR-based detection methods, and we discuss potentials and challenges in using such assays.

A comparison of the accuracy of mushroom identification applications using digital photographs

Objective To compare the accuracy of three popular mushroom identification software applications in identifying mushrooms involved in exposures reported to the Victorian Poisons Information Centre and Royal Botanic Gardens Victoria. Background Over the past 10 years, an increasing number of software applications have been developed for use on smart phones and tablet devices to identify mushrooms. We have observed an increase in poisonings after incorrect identification of poisonous species as edible, using these applications. Design We compared the accuracy of three iPhone™ and Android™ mushroom identification applications: Picture Mushroom (Next Vision Limited©), Mushroom Identificator (Pierre Semedard©), and iNaturalist (iNaturalist, California Academy of Sciences©). Each app was tested independently by three researchers using digital photographs of 78 specimens sent to the Victorian Poisons Information Centre and Royal Botanic Gardens Victoria over a two-year period, 2020–2021. Mushroom identification was confirmed by an expert mycologist. For each app, individual and combined results were compared. Results Picture Mushroom was the most accurate of the three apps and correctly identified 49% (95% CI [0–100]) of specimens, compared with Mushroom Identificator (35% [15–56]) and iNaturalist (35% [0–76]). Picture Mushroom correctly identified 44% of poisonous mushrooms [0–95], compared with Mushroom Identificator (30% [1–58]) and iNaturalist (40% [0–84), but Mushroom Identificator identified more specimens of Amanita phalloides correctly (67%), compared to Picture Mushroom (60%) and iNaturalist (27%). Amanita phalloides was falsely identified, twice by Picture Mushroom and once by iNaturalist. Conclusions Mushroom identification applications may be useful future tools to assist clinical toxicologists and the general public in the accurate identification of mushrooms species but, at present, are not reliable enough to exclude exposure to potentially poisonous mushrooms when used alone.

Https://gmsmushrooms.org/: A comprehensive Online Database of Mushrooms in the Greater Mekong Sub Region

The Greater Mekong Sub-region (GMS) is a global biodiversity hotspot and habitat for many ecologically and economically important mushrooms species. The diversity of mushrooms in the GMS has been studied and published in different journals, and many have been documented from different countries in the GMS region but data remain mostly fragmented. A comprehensive website, https://gmsmushrooms.org/, was established to update the taxonomy, illustrations, descriptions and other useful information for mushroom species reported from the GMS into a single platform. The website is managed by expert mycologists who regularly update information as new macrofungi are introduced from the GMS.

Clinical recovery of 5 dogs from amatoxin mushroom poisoning using an adapted Santa Cruz protocol for people

To describe the clinical course, treatment, and outcome of 5 dogs following ingestion of toxic Amanita spp. mushrooms containing amatoxins using an adapted version of the Santa Cruz protocol developed for people. Five dogs were presented with clinical signs compatible with amanitin toxicity with witnessed ingestion noted in 3 of 5 dogs. Clinical findings included acute onset vomiting and diarrhea, lethargy, and hepatopathy including signs of fulminant hepatic failure (increased liver enzyme activities, hyperbilirubinemia, prolonged clotting times, and hypoglycemia were noted among these cases). Urine toxicological screening confirmed the presence of Amanita toxins in 4 cases with expert mycologist speciation in the fifth. Core interventions included percutaneous biliary drainage, use of octreotide, and early nil per os orders. All dogs survived to discharge with this treatment strategy. This case series describes the use of a modified version of the Santa Cruz protocol to address amatoxin-induced fulminant hepatic failure in dogs. The protocol was safe, well tolerated, and all patients made a full clinical recovery.

Molecular Identification of Mushroom Species in Italy: An Ongoing Project Aimed at Reinforcing the Control Measures of an Increasingly Appreciated Sustainable Food

Proper investment in mushroom production (farming and wild mushroom picking activities) may represent a winning strategy for many countries, including Italy, to better face the problems of food security and environmental impact, and to break away from imports, enhancing the local products. However, the risk related to the consumption of poisoning species requires governments to implement or reinforce effective control measures to protect consumers. Mushroom identification by phenotype observation is hardly applicable if morphologically-similar species, non-whole specimens, or clinical samples are involved. Genotypic analysis is a valid alternative. An ongoing research project involving the Experimental Zooprophylactic Institute of Lazio and Tuscany, the regional Mycological Inspectorate, the Tuscany Mycological Groups Association, and the Department of Veterinary Sciences of the University of Pisa aims to reinforce the collaboration among institutions for the management of mushroom poisoning. The core’s project aims to develop an internal genetic database to support the identification of wild and cultivated mushroom species in the Italian territory. The database will include Internal Transcribed Spacer (ITS) sequences retrieved from official databases (the NCBI GenBank and the BOLD system) which are considered to be reliable, after a proper selection process, and sequences from specimens collected directly and identified by expert mycologists. Once it is validated, the database will be available and further implementable by the official network of national laboratories.

Wild mushrooms poisoning: a public health challenge

Abstract Issue Mushrooms are considered as ingredient of gourmet cuisine across the globe and mushroom poisonings can occur because of misidentification of a poisonous species as edible. Of the vast number of mushroom species, approximately 100 are toxic(**). Description of the Problem Mushroom poisoning is a public health problem. People should be educated about this poison as often there are no guidelines about the treatment of mushroom toxicity. Despite treatment, the prognosis can be mortal. In the Department of Hygiene and Health Prevention (ATS Val Padana, Italy) expert mycologists offer their consultation free of charge to people who pick mushrooms and register, for each consultation, quantity and toxicity of collected mushrooms. Results In 2018-19, 279 consultations were offered: 124(44.44%) in 2018, 155 (55.56%) in 2019. 58.42% of mushrooms were considered not dangerous for human health, without any difference among the two considered years (p = 0.167). 233.7 kg mushrooms were considered edible (mean 2.71, SD 16.44), 92.95 kg were considered dangerous and destroyed (mean 1.39, SD 6.99). The most identified species were: Agrocybe aegerita (11.47%), Armillaria Mellea (29.75%), Boletus edulis (6.45%), Macrolepiota venenata** (1.43%, N = 4), Xerocomus badius (2.51%), Lyophillum loricatum (1.79%), Agaricus Xanthodermus** (3.23%, N = 9). Intoxications occurred among people who did not refer to our mycology unit. Conclusions Mushroom poisoning represents a major health risk and as it's often consequence of misidentification, a free of charge consultation offered by experts is fundamental to avoid intoxications or deaths. Key messages Mushroom poisoning presents a major health risk. A free of charge consultation offered by experts is fundamental to avoid intoxications or deaths.

Identification of dermatophytes by MALDI-TOF MS technology in the clinical laboratory

Conventional dermatophyte diagnosis includes microscopic and macroscopic characterization of the fungi by a skilled mycologist following fungi cultivation for several weeks. In recent years, matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS) has become useful for highly specific and sensitive identification of bacteria and yeasts from clinical samples. The current study aimed to test the influence of using a protein extraction protocol on dermatophyte identification by MALDI-TOF MS.We collected scrapings of affected skin from forty-six patients diagnosed with tinea. Each sample was cultured on four tubes of Sabouraud Dextrose Agar. After 21–28 days of incubation, fungi morphology was examined by an expert mycologist according to standard microscopic and macroscopic colony characteristics.Dermatophyte identification by MS was performed following 14 (Protocol I) or 21 (Protocol II) days of incubation with direct deposit and addition of formic acid or following an Ethanol/formic acid extraction – "EtOH/FA complete extraction protocol" after 14 days (Protocol III). We found that the EtOH/FA complete extraction protocol had a 100% correct identification rate. While identification to species level was obtained in 0% and 22% of the dermatophytes following Protocol I and Protocol II, respectively, the EtOH/FA complete extraction protocol resulted in 65% identification to the species level. Considering the straightforward performance of the EtOH/FA complete extraction protocol and the high accuracy, we recommend its implementation in routine diagnosis in clinical laboratories.

FunGene-DB: A web-based tool for Polyporales strains authentication

Polyporales are extensively studied wood-decaying fungi with applications in white and green biotechnologies and in medicinal chemistry. We developed an open-access, user-friendly, bioinformatics tool named FunGene-DB (http://www.fungene-db.org). The goal was to facilitate the molecular authentication of Polyporales strains and fruit-bodies, otherwise subjected to morphological studies. This tool includes a curated database that contains ITS1-5.8S-ITS2 rDNA genes screened through a semi-automated pipeline from the International Nucleotide Sequence Database (INSD), and the similarity search BLASTn program. Today, the web-accessible database compiles 2379 accepted sequences, among which 386 were selected as reference sequences (most often fully identified ITS sequences for which a voucher, strain or specimen, has been deposited in a public-access collection). The restriction of the database to one reference sequence per species (or per clade for species complex) allowed most often unequivocal analysis. We conclude that FunGene-DB is a promising tool for molecular authentication of Polyporales. It should be especially useful for scientists who are not expert mycologists but who need to check the identity of strains (e.g. for culture collections, for applied microbiology).

Category Learning Increases Discriminability of Relevant Object Dimensions in Visual Cortex

Learning to categorize objects can transform how they are perceived, causing relevant perceptual dimensions predictive of object category to become enhanced. For example, an expert mycologist might become attuned to species-specific patterns of spacing between mushroom gills but learn to ignore cap textures attributable to varying environmental conditions. These selective changes in perception can persist beyond the act of categorizing objects and influence our ability to discriminate between them. Using functional magnetic resonance imaging adaptation, we demonstrate that such category-specific perceptual enhancements are associated with changes in the neural discriminability of object representations in visual cortex. Regions within the anterior fusiform gyrus became more sensitive to small variations in shape that were relevant during prior category learning. In addition, extrastriate occipital areas showed heightened sensitivity to small variations in shape that spanned the category boundary. Visual representations in cortex, just like our perception, are sensitive to an object's history of categorization.

Mushroom poisoning from species of genus Inocybe (fiber head mushroom): a case series with exact species identification

Background. Many species of the genus Inocybe (family Cortinariaceae, higher Basidiomycetes) are muscarine-containing mycorrhizal mushrooms, ubiquitous around the world. The few published reports on the poisonous Inocybe mushrooms are often limited by the inadequate identification of the species. The clinical course of patients with typical muscarinic manifestations, in whom Inocybe spp. was unequivocally identified, is reported. Case series. Between November 2006 and January 2008 14 consecutive patients with typical muscarinic syndrome after mushroom ingestion were recorded. The clinical manifestations included combinations of nausea, vomiting, diarrhea, abdominal pain, hypersalivation, diaphoresis, tachycardia, bradycardia, hypotension, lacrimation, blurred vision, miosis, tremor, restlessness, flushing, and syncope. Time to onset of toxicity ranged between 15 min and 2 h after consumption, 5 h in one patient. Treatment was supportive, including intravenous fluids, antiemetics, and 1 mg atropine intravenously. Full recovery ensued within 12 h. In all the cases, an expert mycologist unequivocally identified the leftovers of the consumed mushrooms as Inocybe fastigiata, Inocybe geophylla, and Inocybe patouillardii. Conclusion. In this case series of patients who ingested identified muscarine-containing mushrooms supportive treatment and atropine resulted in recovery in all cases.

Mushroom toxins yield to analysis

Mushrooms deserve their notoriety for producing peculiar poisons. Recently, several previously undescribed mushroom toxins, including one family of compounds that's not dangerous to man and another compound that caused a small outbreak of poisonings and several fatalities in Poland during the 1950's, have yielded to chemical analysis. The task apparently isn't easy. According to one chemist who is an avid amateur mycologist, the challenge of doing good chemistry on rare mushrooms often results in a mismatch of talents. Few chemists know much about mushrooms, and so their analyses often may be wasted on improperly identified specimens. And the reverse case creates its own set of problems wherein expert mycologists find themselves at a loss when fine chemical analysis is needed. Fortunately there are exceptions to these rules. And the recent (and long-term) efforts of Theodor Wieland, Heinz Faulstich, and their colleagues at the Max Planck Institute for Medical Research in Heidelberg, West Germany, to analy...

Mycologist Helpful in Mold Allergy Diagnosis

The services of an expert mycologist can be invaluable in the diagnosis of mold allergy, according to Mayer A. Green, MD, of Pittsburgh, who addressed the American College of Allergy meeting in Bal Harbour, Fla, Mar 1-6. When faced with a suspected case of mold allergy, it is essential that the physician, in addition to making mold sensitization studies, know the atmospheric mold content of the area in which the patient resides. At this point, the mycologist's services are useful, he said. In the United States, reference is often made to the so-called atmospheric X factors, which do not coincide with identifiable pollens. "The probability is that some of the seasonal and post-seasonal respiratory allergic symptoms may be due to molds." Only recently, he said, have allergists been made aware of the role of molds in producing allergies. Much of the identification and classification of molds has been carried out

An Addition to the Parasites of the Human Ear

long prevalent in the groups to which the ear-parasites belong, recently abated in the Black Moulds by A. Fischer, but still in force in the Imperfect Fungi, and the fact that otologists have not been much concerned with mycology nor always able to command the assistance of expert mycologists, have united with the inherent difficulties due to the effect of situs on the forms themselves to produce errors in determination, uncertainty, and even controversy. The greater number of the species reported and the greater number of the well identified and authenticated species belong to the Aspergillaceae. Next come the Mucoraceae. The Protoascineae and Pezizineae contribute one each. In addition there are several