A new endophytic species of Microthecium (Melanosporales, Sordariomycetes, Pezizomycotina, Ascomycota) from Mallorca (Balearic Islands, Spain) / Una nueva especie endofítica de Microthecium (Melanosporales, Sordariomycetes, Pezizomycotina, Ascomycota) de Mallorca (Islas Baleares, España)

Background The genus Microthecium contains 31 species worldwide distributed. Most of them are saprobic on soil and plant debris, but a few have been reported as mycoparasites on hypocrealean fungi. By contrast, this genus has never been reported as phytopathogenic, nor endophytic. Aims To isolate and identify endophytic fungi from Mediterranean herbaceous plants and trees in order to contribute to the knowledge of the hosts and their geographical location. The present work has been focused on the study of endophytic fungi of hawthorn (Crataegus monogyna). Methods The following steps were taken: i, isolation of the fungal strain from living stems of C. monogyna; ii, cultural and micro-morphological study, and iii, sequence comparison of different genetic markers by BLAST search with sequences deposited in GenBank. Results At the present work we describe a new species of the genus, Microthecium pleomorphosporum, isolated from living stems of C. monogyna in Mallorca (Balearic Islands, Spain). This fungus is characterized by the production of non-ostiolate perithecia and two sorts of ascospores (some smooth-walled, others delicately reticulated) bearing a germ pore at each end which are frequently ornamented by a surrounding donut-like structures, and a phialidic asexual morph and bulbils. The morphologically closest related species is Microthecium tenuissimum, which has bigger ascospores and lacks asexual reproduction. Phylogenetically, M pleomorphosporum is close-related to other species of the genus, although no genetic marker that discriminates this new species from other phylogenetically closer ones could be elucidated as a gold standard. Conclusions M. pleomorphosporum, order Melanosporales, is reported here as the first endophytic species of C. monogyna. (AU)

Antecedentes El género Microthecium incluye 31 especies de distribución cosmopolita. La gran mayoría de ellas son saprobias en el suelo y se encuentran sobre detritos vegetales, y tan solo unas pocas se han descrito como micoparásitas de hongos hipocreales. Por el contrario, nunca se había descrito a este género como fitopatógeno ni endofítico. Objetivos Aislar e identificar hongos endófitos de plantas herbáceas y árboles mediterráneos con el fin de contribuir al conocimiento de los hospederos y su situación geográfica. El presente trabajo se ha centrado en el estudio de los hongos endófitos del espino albar (Crataegus monogyna). Métodos Se incluyeron diferentes procesos: (1) aislamiento de la cepa fúngica a partir de tallos vivos de Crataegus monogyna; (2) estudio cultural y micromorfológico y (3) comparación de las secuencias nucleotídicas de diferentes marcadores filogenéticamente informativos con secuencias de taxones conocidos depositadas en el GenBank mediante búsqueda BLAST. Resultados En el presente trabajo describimos una nueva especie para el género, Microthecium pleomorphosporum, aislada de tallos vivos de C. monogyna en Mallorca (Islas Baleares, España). Este hongo se caracteriza por la producción de peritecios no ostiolados y de dos tipos de ascosporas (unas con paredes lisas y otras con paredes delicadamente reticuladas), con un poro germinativo en cada extremo frecuentemente ornamentado por estructuras circundantes en forma de rosquilla, y un estado asexual que produce conidios fialídicos y bulbillos. La especie morfológicamente más cercana es Microthecium tenuissimum, que tiene ascosporas más grandes y carece de multiplicación asexual. Filogenéticamente, Microthecium pleomorphosporum está estrechamente emparentada con otras especies del género y no se ha podido establecer ningún marcador genético de referencia (gold standard) para discriminar esta nueva especie de otras filogenéticamente más próximas. Conclusiones ... (AU)

A new endophytic species of Microthecium (Melanosporales, Sordariomycetes, Pezizomycotina, Ascomycota) from Mallorca (Balearic Islands, Spain).

BACKGROUND: The genus Microthecium contains 31 species worldwide distributed. Most of them are saprobic on soil and plant debris, but a few have been reported as mycoparasites on hypocrealean fungi. By contrast, this genus has never been reported as phytopathogenic, nor endophytic. AIMS: To isolate and identify endophytic fungi from Mediterranean herbaceous plants and trees in order to contribute to the knowledge of the hosts and their geographical location. The present work has been focused on the study of endophytic fungi of hawthorn (Crataegus monogyna). METHODS: The following steps were taken: i, isolation of the fungal strain from living stems of C. monogyna; ii, cultural and micro-morphological study, and iii, sequence comparison of different genetic markers by BLAST search with sequences deposited in GenBank. RESULTS: At the present work we describe a new species of the genus, Microthecium pleomorphosporum, isolated from living stems of C. monogyna in Mallorca (Balearic Islands, Spain). This fungus is characterized by the production of non-ostiolate perithecia and two sorts of ascospores (some smooth-walled, others delicately reticulated) bearing a germ pore at each end which are frequently ornamented by a surrounding donut-like structures, and a phialidic asexual morph and bulbils. The morphologically closest related species is Microthecium tenuissimum, which has bigger ascospores and lacks asexual reproduction. Phylogenetically, M pleomorphosporum is close-related to other species of the genus, although no genetic marker that discriminates this new species from other phylogenetically closer ones could be elucidated as a gold standard. CONCLUSIONS: M. pleomorphosporum, order Melanosporales, is reported here as the first endophytic species of C. monogyna.

A revision of malbranchea-like fungi from clinical specimens in the United States of America reveals unexpected novelty.

The fungi of the order Onygenales can cause important human infections; however, their taxonomy and worldwide occurrence is still little known. We have studied and identified a representative number of clinical fungi belonging to that order from a reference laboratory in the USA. A total of 22 strains isolated from respiratory tract (40%) and human skin and nails (27.2%) showed a malbranchea-like morphology. Six genera were phenotypically and molecularly identified, i.e. Auxarthron/Malbranchea (68.2%), Arachnomyces (9.1%), Spiromastigoides (9.1%), and Currahmyces (4.5%), and two newly proposed genera (4.5% each). Based on the results of the phylogenetic study, we synonymized Auxarthron with Malbranchea, and erected two new genera: Pseudoarthropsis and Pseudomalbranchea. New species proposed are: Arachnomyces bostrychodes, A. graciliformis, Currahmyces sparsispora, Malbranchea gymnoascoides, M. multiseptata, M. stricta, Pseudoarthropsis crassispora, Pseudomalbranchea gemmata, and Spiromastigoides geomycoides, along with a new combination for Malbranchea gypsea. The echinocandins showed the highest in vitro antifungal activity against the studied isolates, followed by terbinafine and posaconazole; in contrast, amphotericin B, fluconazole, itraconazole and 5-fluorocytosine were less active or lacked in vitro activity against these fungi.

Three New Derivatives of Zopfinol from Pseudorhypophila Mangenotii gen. et comb. nov.

Triangularia mangenotti was analyzed for the production of secondary metabolites, resulting in the isolation of known zopfinol (1) and its new derivatives zopfinol B-C (2-4), the 10-membered lactones 7-O-acetylmultiplolide A (5) and 8-O-acetylmultiplolide A (6), together with sordarin (7), sordarin B (8), and hypoxysordarin (9). The absolute configuration of 1 was elucidated by the synthesis of MPTA-esters. Compound 1 showed antimicrobial activity against the Gram-positive bacteria Bacillus subtilis and Staphylococcus aureus and the fungus Mucor hiemalis. While 4 was weakly antibacterial, 3 showed stronger antibiotic activity against the Gram-positive bacteria and weak antifungal activity against M. hiemalis and Rhodotorula glutinis. We furthermore observed the cytotoxicity of 1, 3 and 4 against the mammalian cell lines KB3.1 and L929. Moreover, the new genus Pseudorhypophila is introduced herein to accommodate Triangularia mangenotii together with several species of Zopfiella-Z. marina, Z. pilifera, and Z. submersa. These taxa formed a well-supported monophyletic clade in the recently introduced family Navicularisporaceae, located far from the type species of the respective original genera, in a phylogram based on the combined dataset sequences of the internal transcribed spacer region (ITS), the nuclear rDNA large subunit (LSU), and fragments of the ribosomal polymerase II subunit 2 (rpb2) and ß-tubulin (tub2) genes. Zopfiella submersa is synonymized with P. marina due to the phylogenetic and morphological similarity. The isolation of zopfinols 1-4 and sordarins 7-9 confirms the potential of this fungal order as producers of bioactive compounds and suggests these compounds as potential chemotaxonomic markers.

Scedosporium spp. from Clinical Setting in Argentina, with the Proposal of the New Pathogenic Species Scedosporium americanum.

Species of the genus Scedosporium (family Microascaceae, phylum Ascomycota) are responsible for a wide range of opportunistic human infections, and have a low susceptibility to most antifungal drugs. It is well known that the pattern of Scedosporium species distribution varies according to geographic region. To assess the diversity of Scedosporium species in Argentina involved in human infections, we carried out a retrospective study reviewing 49 strains from clinical samples sent for diagnosis to the National Clinical Mycology Reference Laboratory between 1985 and 2019. Then, a phenotypic characterization, a phylogenetic study and and in vitro susceptibility test to antifungals were carried out. An analysis of combined nucleotide sequences dataset of the internal transcribed spacer of the ribosomal DNA (ITS) and of a fragment of the ß-tubulin gene (BT2) demonstrated that 92 % of the strains belonged to the species S. boydii, S. apiospermum and S. angustum, all them pertaining to S. apiospermum species complex. However, two strains (4%) were identified as S. aurantiacum, a species never reported in clinical settings in the Americas'. Surprisingly, one of them displayed a polycytella-like conidiogenesis, up to date only reported for S. apiospermum. In addition, the strain DMic 165285 was phylogenetically located far away from the rest of the species, so is proposed as the novel species Scedosporium americanum. On the other hand, from all seven antifungals tested, voriconazole and posaconazole were the most active drugs against Scedosporium spp.

New Xerophilic Species of Penicillium from Soil.

Soil is one of the main reservoirs of fungi. The aim of this study was to study the richness of ascomycetes in a set of soil samples from Mexico and Spain. Fungi were isolated after 2% w/v phenol treatment of samples. In that way, several strains of the genus Penicillium were recovered. A phylogenetic analysis based on internal transcribed spacer (ITS), beta-tubulin (BenA), calmodulin (CaM), and RNA polymerase II subunit 2 gene (rpb2) sequences showed that four of these strains had not been described before. Penicillium melanosporum produces monoverticillate conidiophores and brownish conidia covered by an ornate brown sheath. Penicillium michoacanense and Penicillium siccitolerans produce sclerotia, and their asexual morph is similar to species in the section Aspergilloides (despite all of them pertaining to section Lanata-Divaricata). P. michoacanense differs from P. siccitolerans in having thick-walled peridial cells (thin-walled in P. siccitolerans). Penicillium sexuale differs from Penicillium cryptum in the section Crypta because it does not produce an asexual morph. Its ascostromata have a peridium composed of thick-walled polygonal cells, and its ascospores are broadly lenticular with two equatorial ridges widely separated by a furrow. All four new species are xerophilic. Despite the genus Penicillium containing more than 480 known species, they are rarely reported as xerophilic.

A new pleosporalean fungus isolated from superficial to deep human clinical specimens.

Human infections by pleosporalean fungi (class Dothideomycetes, phylum Ascomycota) are rarely reported. Because their identification is challenging using morphological characterization, several phylogenetic markers must be sequenced for an accurate identification and taxonomical placement of the isolates. Three isolates of clinical origin were phenotypically characterized, but due to the absence of relevant morphological traits, D1-D2 domains of the 28S nrRNA gene (LSU), the internal transcribed spacer region (ITS) of the nrRNA, and fragments of the RNA polymerase II subunit 2 (rpb2) and translation elongation factor 1-alpha (tef1) genes were sequenced to allow a phylogenetic analysis that would solve their phylogenetic placement. That analysis revealed that these isolates did not match any previously known pleosporalean genera, and they are proposed here as the new fungal genus, Gambiomyces. Unfortunately, the isolates remained sterile, which, consequently, made the morphological description of the reproductive structures impossible. Future studies should try to understand the behaviour of this fungus in nature as well as its characteristics as an opportunistic fungal pathogen. Molecular identification is becoming an essential tool for proper identification of Dothideomycetes of clinical origin. LAY ABSTRACT: We describe a new pleosporalen pathogenic fungus, Gambiomyces profunda, found in superficial to deep samples from a human patient. Because all strains remained sterile, the fungus was finally identified following a phylogenetic analysis by using four different molecular markers.

Seven New Cytotoxic and Antimicrobial Xanthoquinodins from Jugulospora vestita.

During the course of a screening for novel biologically active secondary metabolites produced by the Sordariomycetes (Ascomycota, Fungi), the ex-type strain of Jugulospora vestita was found to produce seven novel xanthone-anthraquinone heterodimers, xanthoquinodin A11 (1) and xanthoquinodins B10-15 (2-7), together with the already known compound xanthoquinodin B4 (8). The structures of the xanthoquinodins were determined by analysis of the nuclear magnetic resonance (NMR) spectroscopic and mass spectrometric data. Moreover, the absolute configurations of these metabolites were established by analysis of the 1H-1H coupling constants, nuclear Overhauser effect spectroscopy (NOESY) correlations, and Electronic Circular Dichroism (ECD) spectroscopic data. Antifungal and antibacterial activities as well as cytotoxicity of all compounds were tested. Xanthoquinodin B11 showed fungicidal activities against Mucor hiemalis [minimum inhibitory concentration (MIC) 2.1 µg/mL], Rhodotorula glutinis (MIC 2.1 µg/mL), and Pichia anomala (MIC 8.3 µg/mL). All the compounds 1-8 displayed anti-Gram-positive bacteria activity (MIC 0.2-8.3 µg/mL). In addition, all these eight compounds showed cytotoxicity against KB 3.1, L929, A549, SK-OV-3, PC-3, A431, and MCF-7 mammalian cell lines. The six novel compounds (1-3, 5-7), together with xanthoquinodin B4, were also found in the screening of other strains belonging to Jugulospora rotula, revealing the potential chemotaxonomic significance of the compound class for the genus.

Re-Evaluation of the Order Sordariales: Delimitation of Lasiosphaeriaceae s. str., and Introduction of the New Families Diplogelasinosporaceae, Naviculisporaceae, and Schizotheciaceae.

The order Sordariales includes the polyphyletic family Lasiosphaeriaceae, which comprises approximately 30 genera characterized by its paraphysate ascomata, asci with apical apparati, and mostly two-celled ascospores, which have a dark apical cell and a hyaline lower cell, frequently ornamented with mucilaginous appendages[...].

New Taxa of the Family Amniculicolaceae (Pleosporales, Dothideomycetes, Ascomycota) from Freshwater Habitats in Spain.

With the exception of the so-called Ingoldian fungi, the diversity and distribution of the freshwater aero-aquatic or facultative fungi are not well known in Spain. In view of that, we collected and placed into wet chambers 105 samples of submerged and decomposing plant debris from various places in Spain, looking for individuals belonging to these latter two morpho-ecological groups of fungi. As a result, we found and isolated in pure culture several fungi, the morphology of some of them belonging to the family Amniculicolaceae (order Pleosporales, class Dothideomycetes). After a careful phenotypic characterization and a phylogenetic tree reconstruction using a concatenated sequence dataset of D1-D2 domains of the 28S nrRNA gene (LSU), the internal transcribed spacer region (ITS) of the nrDNA, and a fragment of the translation elongation factor 1-alpha (tef1) gene, we report the finding of three new species of the genus Murispora: Murispora navicularispora, which produces cinnamon-colored, broadly fusiform to navicular ascospores; Murispora fissilispora, which has as a remarkable characteristic the production of both sexual and asexual morphs in vitro; and Murispora asexualis, the unique species of the genus that lacks a sexual morph. As a consequence of the phylogenetic study, we introduce the new aero-aquatic genus Fouskomenomyces, with a new combination (Fouskomenomyces cupreorufescens, formerly Spirosphaera cupreorufescens as the type species of the genus) and a new species, Fouskomenomyces mimiticus; we propose the new combinations Murispora bromicola (formerly Pseudomassariosphaeria bromicola) and Murispora triseptata (formerly Pseudomassariosphaeria triseptata); and we resurrect Massariosphaeria grandispora, which is transferred to the family Lopiostomataceae.

Structure elucidation and absolute configuration of metabolites from the soil-derived fungus Dictyosporium digitatum using spectroscopic and computational methods.

Following the discovery of a new class of compounds that inhibit the mucosa-associated lymphoid tissue lymphoma translocation 1 (MALT1) protease in a prior study, further chemical investigation of the Dictyosporium digitatum fungus resulted in the identification of 16 additional metabolites, including 12 undescribed compounds (1-12). The constitution and relative configuration of these new molecules were established by comprehensive NMR and HRMS analyses. Their absolute configurations were determined by employing Mosher's ester analysis and TDDFT ECD calculations. Two sesquiterpenes, dictyosporins A (1) and B (2), possess an undescribed eudesmen-type of structural scaffold. The ability of the isolated compounds to inhibit MALT1 proteolytic activity was evaluated, but none of them exhibited significant inhibition.

Fungal Diversity of Deteriorated Sparkling Wine and Cork Stoppers in Catalonia, Spain.

Filamentous fungi are rarely reported as responsible for spoiling wine. Cork taint was detected in sparkling wine; therefore, we investigated fungal contamination as a possible cause of organoleptic alteration. Spoiled wine was filtered and membranes were plated onto potato dextrose agar (PDA). The cork stoppers used for sealing bottles were cut and also plated onto PDA. Fungal strains were phenotypically characterized and molecularly identified by sequencing of a fragment of the 28S nrRNA gene (LSU) and (occasionally) by other additional molecular markers. Twenty-seven strains were isolated and sixteen species were identified, all of them belonging to the phylum Ascomycota. The fungi isolated from wine were three species of Aspergillus section Nidulantes, a species of Penicillium section Exicaulis and Beauveria bassiana. Candida patagonica was isolated from both sort of samples, and the fungi isolated from cork stoppers were Altenaria alternata and Cladosporium cladosporioides. Surprisingly, most of the taxa recovered from the cork stoppers and/or wine were new to the science: a new genus (Dactylodendron) and seven new species belonging to the genera Cladophialophora, Dactylodendron, Kirschsteiniothelia, Rasamsonia, and Talaromyces. Future studies could let us know if these fungi would be able to produce compounds responsible for cork taint.

Two new species of Gloniopsis (Hysteriales, Ascomycota) from clinical specimens: Morphological and molecular characterisation.

BACKGROUND: The coelomycetes comprise a wide range of fungal species distributed in at least three different classes of the phylum Ascomycota. These are morphologically characterised by producing their conidia inside of fruiting bodies called pycnidia or acervuli, and only a reduced number of species are able to cause human infections. However, their identification in the clinical laboratory is often difficult, due to their few morphological features or because they remain sterile. MATERIALS AND METHODS: In the present study, three isolates of coelomycetes of clinical origin were phenotypically and molecularly studied, by sequencing the D1-D2 fragment of the 28S nuclear ribosomal RNA (nrRNA) (LSU), the internal transcribed spacer region (ITS1-5.8S-ITS2) and a fragment of the translation elongation factor 1-alpha (tef1) genes. RESULTS AND CONCLUSIONS: As result of the molecular analysis, the isolates were identified as belonging to the genus Gloniopsis (order Hysteriales, Dothideomycetes) but without the characteristics of any of the species described so far. Therefore, we propose the new species Gloniopsis percutanea and Gloniopsis pneumoniae. Furthermore, this study revealed that some isolates from clinical specimens identified previously as Rhytidhysteron spp. were misidentified, and considering the few studies in the order Hysteriales and the scarce number of sequences of phylogenetic markers, future revisions of this order should be performed to clarify their taxonomy and obtain a better identification from isolates involved in human mycoses.

Diversity of coelomycetous fungi in human infections: A 10-y experience of two European reference centres.

The coelomycetous fungi are difficult to properly identify from their phenotypic characterization and their role as etiologic agents of human infections is not clear. We studied the species distribution of these fungi among clinical isolates that had been collected and stored over a ten-year period in two European reference laboratories (France and Spain). We identified phenotypically and molecularly 97 isolates by sequencing the D1-D2 fragment of the 28S nrRNA (LSU) gene and we provided the in vitro antifungal susceptibility pattern of seven antifungals against 46 isolates. Species of the orders Pleosporales and Glomerellales were present in both collections, and Botryosphaeriales and Diaporthales only in the French one. The most prevalent species were Medicopsis romeroi, Neocucurbitaria keratinophila, Neocucurbitaria unguis-hominis and Paraconiothyrium cyclothyrioides, which had been recovered primarily from superficial tissues. The Didymellaceae was the most common family represented, with 27 isolates distributed into five genera. Most of the isolates tested were susceptible to antifungals, and only the geometric mean (GM) and minimal inhibitory concentration (MIC) values of itraconazole and caspofungin had higher values. This study provides a good picture of the great diversity of coelomycetous fungi in the European clinical context, and the basis for future studies on this interesting but neglected group of fungi.

Secondary Metabolites from the Fungus Dictyosporium sp. and Their MALT1 Inhibitory Activities.

Bioassay-guided separation of an extract from a Dictyosporium sp. isolate led to the identification of six new compounds, 1-6, together with five known compounds, 7-11. The structures of the new compounds were primarily established by extensive 1D and 2D NMR experiments. The absolute configurations of compounds 3-6 were determined by comparison of their experimental electronic circular dichroism (ECD) spectra with DFT quantum mechanical calculated ECD spectra. Compounds 3-5 possess novel structural scaffolds, and biochemical studies revealed that oxepinochromenones 1 and 7 inhibited the activity of MALT1 protease.

Novel Paranannizziopsis species in a Wagler's viper (Tropidolaemus wagleri), tentacled snakes (Erpeton tentaculatum), and a rhinoceros snake (Rhynchophis boulengeri) in a zoological collection.

We report several cases of fungal infections in snakes associated with a new species within the genus Paranannizziopsis. Three juvenile Wagler's vipers (Tropidolaemus wagleri) presented with skin abnormalities or ulcerative dermatitis, and two snakes died. Histologic examination of skin from the living viper revealed hyperplastic, hyperkeratotic, and crusting epidermitis with intralesional fungal elements. The terrestrial Wagler's vipers were housed in a room with fully aquatic tentacled snakes (Erpeton tentaculatum), among which there had been a history of intermittent skin lesions. Approximately 2 months after the biopsy of the viper, a skin sample was collected from one tentacled snake (TS1) with skin abnormalities and revealed a fungal infection with a similar histologic appearance. Fungal isolates were obtained via culture from the Wagler's viper and TS1 and revealed a novel species, Paranannizziopsis tardicrescens, based on phenotypic characterization and molecular analysis. P. tardicrescens was cultured and identified by DNA sequence analysis 8 months later from a dead tentacled snake in an exhibit in an adjacent hallway and 13 months later from a living rhinoceros snake (Rhynchophis boulengeri) with two focal skin lesions. Antifungal susceptibility testing on three of four cultured isolates demonstrated potent in vitro activity for terbinafine and voriconazole.

Neocucurbitaria keratinophila: An emerging opportunistic fungus causing superficial mycosis in Spain.

Although there have been few reports of opportunistic infections (superficial and systemic) caused by coelomycetous fungi, they are becoming more frequent. Neocucurbitaria keratinophila (formerly Pyrenochaeta keratinophila), characterized by producing pycnidial conidiomata and small hyaline conidia, seems to be an emergent opportunistic pathogen in Spain. Since this fungus was first reported from human keratitis, eight strains have been isolates from clinical cases in Spain. This is a retrospective study of these fungal strains, including phenotypic and molecular characterizations, and in vitro antifungal susceptibility assays. These clinical strains were identified by sequencing four phylogenetic markers such as the internal transcribed spacer region (ITS1-5.8S-ITS2) and fragments of the 28S nrRNA (LSU), beta-tubulin (tub2), and RNA polymerase II subunit 2 (rpb2) genes, and by morphological characterization. All the strains tested were susceptible to the majority of antifungals, being isavuconazole the only drug that showed a poor antifungal activity.

Mucormycosis: Battle with the Deadly Enemy over a Five-Year Period in India.

Mucormycosis is an emerging opportunistic fungal infection. Increasing immunocompromization, widespread use of antibacterial and antifungal agents (such as voriconazole prophylaxis), carcinomas, transplantation and lifestyle diseases such as diabetes are the main contributors to this situation. The predominant clinical manifestations of mucormycosis vary from host to host, with rhino-orbital-cerebral, pulmonary, cutaneous, and gastrointestinal infections being the most common. In India, the prevalence of mucormycosis is approximately 0.14 cases/1000 population, which is about 70 times the worldwide-estimated rate for mucormycosis. The present study was undertaken over a period of five years (January 2009-December 2014) to determine the prevalence of mucormycosis. The samples suspected of mucormycosis were examined by direct KOH wet mount and cultured on Sabouraud's dextrose agar without actidione and on blood agar as per standard mycological techniques. Histopathological correlation was done for most of the cases. Antifungal susceptibility testing was performed by the EUCAST reference method. We identified a total of 82 cases of mucormycosis out of a total of 6365 samples received for mycological culture and examination during the said time period. Out of these, 56 were male patients and 27 were females. Most common presentation was rhino-orbito-cerebral (37), followed by cutaneous (25), pulmonary (14), oral cavity involvement (4) and gastrointestinal (2). The most common risk factors were diabetes and intramuscular injections. The fungi isolated were Rhizopus arrhizus (17), Apophysomyces variabilis (12), R. microsporus (9), Lichtheimia ramosa (8), Saksenaea erythrospora (5), Syncephalastrum racemosus (4), R. homothallicus (2), Rhizomucor pusillus (1), Mucor irregularis (1) and A. elegans (1). The mainstay of the treatment was amphotericin B, along with extensive surgical debridement whenever feasible. Most of the patients (50) recovered, but 25 died. The rest of the patients left against medical advice. "Nip in the Bud" should be the mantra for clinicians/surgeons for a favorable prognosis. Early diagnosis, prompt institution of appropriate antifungal therapy, surgical debridement whenever necessary, knowledge of risk factors and their timely reversal is the key for management.