Zygomycete Fungus Research Articles

Metabolic engineering of Mucor circinelloides to improve astaxanthin production.

Astaxanthin is a bioactive natural pigment with antioxidant properties. It has extensive applications within the industrial sector as well as in human and animal health. Mucor circinelloides is a zygomycete fungus that accumulates β-carotene as the main carotenoid compound. M. circinelloides is a well-known model organism among Mucorales for studying carotenogenesis in fungi, which makes it a promising candidate for the biotechnological production of carotenoids. In this study, β-carotene hydroxylase (crtR-B) and ketolase (bkt) genes (codon-optimized) were coexpressed from Haematococcus pluvialis in M. circinelloides using two potent promoters gpd1 and zrt1 respectively to generate an astaxanthin-producing biofactory. Following 72h of cultivation, the recombinant M. circinelloides Mc-57 obtained in this study produced 135 ± 8µg/g of astaxanthin. This is the highest reported amount in M. circinelloides to date. The mRNA levels of crtR-B and bkt in Mc-57 were assayed using RT-qPCR. These levels showed a 5.7-fold increase at 72h and a 5.5-fold increase at 24h, respectively, compared to the control strain. This demonstrated the successful overexpression of both genes, which correlated with the production of astaxanthin in the Mc-57. Moreover, the addition of glutamate (2g/L) and mevalonate (15 mM) resulted in an increase in astaxanthin production in the recombinant strain. The results showed that the combined addition of these metabolic precursors resulted in 281 ± 20µg/g of astaxanthin, which is 2.08-fold higher than the control medium (135 ± 8µg/g). The addition of metabolic precursors also positively impacted the biomass growth of Mc-57, reaching 11.2 ± 0.57g/L compared to 9.1 ± 0.23g/L (control medium). The study successfully addressed the challenge of balancing the accumulation of astaxanthin with biomass growth, which has been regarded as common bottleneck in the metabolic engineering of microbial cells. The development of a recombinant fungal strain of M. circinelloides not only increased astaxanthin content. Additionally, it provided a foundation for further improvement of the biotechnological production of astaxanthin in M. circinelloides.

Fungi of the Arctic Seas

The abundance and diversity of mycelial fungi in the bottom sediments of the Arctic Ocean seas (the Greenland, Barents and Kara seas) were studied. Samples of the surface bottom sediments were collected during the 84th (July‒August 2021) and 86th (October‒November 2021) cruises of RV Akademik Mstislav Keldysh. The taxonomic affiliation of the isolated fungi was determined using polyphasic taxonomy. The isolated fungi belonged to 16 genera of different classes of ascomycetous, basidiomycetous, and zygomycetous fungi. The effect of cultivation temperature and different NaCl concentrations on fungal growth was determined, as well as the effect of cultivation conditions on the fatty acid profile for the strains capable of growth on media with increased osmotic potential. While fatty acid composition was shown to be affected by changes in environmental conditions, the response to osmotic stress differed among the studied cultures from deep-sea sediments.

Combating Black Fungus: Using Allicin as a Potent Antifungal Agent against Mucorales.

Invasive fungal (IF) diseases are a leading global cause of mortality, particularly among immunocompromised individuals. The SARS-CoV-2 pandemic further exacerbated this scenario, intensifying comorbid IF infections such as mucormycoses of the nasopharynx. In the work reported here, it is shown that zygomycetes, significant contributors to mycoses, are sensitive to the natural product allicin. Inhibition of Mucorales fungi by allicin in solution and by allicin vapor was demonstrated. Mathematical modeling showed that the efficacy of allicin vapor is comparable to direct contact with the commercially available antifungal agent amphotericin B (ampB). Furthermore, the study revealed a synergistic interaction between allicin and the non-volatile ampB. The toxicity of allicin solution to human cell lines was evaluated and it was found that the half maximal effective concentration (EC50) of allicin was 25-72 times higher in the cell lines as compared to the fungal spores. Fungal allicin sensitivity depends on the spore concentration, as demonstrated in a drop test. This study shows the potential of allicin, a sulfur-containing defense compound from garlic, to combat zygomycete fungi. The findings underscore allicin's promise for applications in infections of the nasopharynx via inhalation, suggesting a novel therapeutic avenue against challenging fungal infections.

Solid-state fermentation with Zygomycetes fungi as a tool for biofortification of apple pomace with γ-linolenic acid, carotenoid pigments and phenolic antioxidants

In the last few years, there has been a growing interest in the more efficient utilization of agricultural and food by-products. Apples are among the most processed fruits in the world that generate huge quantities of processing waste biomasses. Therefore, the objective of this study was to improve the nutritional value of apple pomaces with γ-linolenic acid (GLA) and carotenoid pigments by solid-state fermentation (SSF) using two Zygomycetes fungi (Actinomucor elegans and Umbelopsis isabellina). The impact of fermentation periods on the polyphenol content and antioxidant capacity of the bioprocessed apple pomace was also investigated. The accumulated lipids were composed primarily of neutral fractions (mostly triacylglycerols). SSF with U. isabellina yielded a 12.72% higher GLA content than with A. elegans (3.85 g GLA/kg DW of pomace). Contrary to the lipogenic capacity, A. elegans showed higher carotenoids and phenolic antioxidants productivity than U. isabellina. The maximum concentrations for β-carotene (433.11 μg/g DW of pomace-SSF with A. elegans and 237.68 μg/g DW of pomace-SSF with U. isabellina), lutein (374.48 μg/g DW- A. elegans and 179.04 μg/g DW- U. isabellina) and zeaxanthin (247.35 μg/g DW- A. elegans and 120.41 μg/g DW- U. isabellina) were registered on the 12th day of SSFs. In the case of SSF with A. elegans, the amount of total phenolics increased significantly (27%) by day 4 from the initial value (2670.38 μg of gallic acid equivalents/g DW) before slowly decreasing for the remaining period of the fungal growth. The experimental findings showed that a prolonged fermentation (between 8 and 12 days) should be applied to obtain value-added apple pomaces (rich in GLA and carotenoids) with potential pharmaceutical and functional food applications. Moreover, the SSF processes of simultaneous bioaccumulation of valuable fatty acids, carotenoids and phenolic antioxidants proposed in the present study may open up new challenges for biotechnological production of industrially important biomolecules using abundant and unexploited apple pomaces.

Sequencing the Genomes of the First Terrestrial Fungal Lineages: What Have We Learned?

The first genome sequenced of a eukaryotic organism was for Saccharomyces cerevisiae, as reported in 1996, but it was more than 10 years before any of the zygomycete fungi, which are the early-diverging terrestrial fungi currently placed in the phyla Mucoromycota and Zoopagomycota, were sequenced. The genome for Rhizopus delemar was completed in 2008; currently, more than 1000 zygomycete genomes have been sequenced. Genomic data from these early-diverging terrestrial fungi revealed deep phylogenetic separation of the two major clades-primarily plant-associated saprotrophic and mycorrhizal Mucoromycota versus the primarily mycoparasitic or animal-associated parasites and commensals in the Zoopagomycota. Genomic studies provide many valuable insights into how these fungi evolved in response to the challenges of living on land, including adaptations to sensing light and gravity, development of hyphal growth, and co-existence with the first terrestrial plants. Genome sequence data have facilitated studies of genome architecture, including a history of genome duplications and horizontal gene transfer events, distribution and organization of mating type loci, rDNA genes and transposable elements, methylation processes, and genes useful for various industrial applications. Pathogenicity genes and specialized secondary metabolites have also been detected in soil saprobes and pathogenic fungi. Novel endosymbiotic bacteria and viruses have been discovered during several zygomycete genome projects. Overall, genomic information has helped to resolve a plethora of research questions, from the placement of zygomycetes on the evolutionary tree of life and in natural ecosystems, to the applied biotechnological and medical questions.

Enhancing feeding value of corn distiller’s grains with solubles via fungal co-cultured solid-state fermentation for monogastric animal nutrition

Corn distiller’s dried grains with solubles (cDDGS) as a major co-product from dry-grind corn-ethanol processes have imbalanced key amino acids (AA), high indigestible fiber and phytate, and risk of mycotoxin contamination, which limit its inclusion rate in monogastric animal diets. This study used cellulolytic fungus Trichoderma reesei (TR), two zygomycete fungi Mucor indicus (MI) and Rizopus oryzae (RO) and their co-cultures to ferment corn wet distiller’s grains with solubles (WDGS) with the supply of soybean hulls and urea via solid-state fermentation at 28 °C for up to 9 days. Compared to control (non-inoculation) and monoculture, TR and RO co-culture (TR+RO) after 6 and 9 days showed more balanced branch-chain AA (BCAA) of valine (Val), leucine (Leu), and isoleucine (Ile), higher phytate reduction (56–61%), lower concentration of deoxynivalenol (DON), zearalenone (ZEN) and total aflatoxins (AFT). However, the treatments with MI (MI, RO+MI, TR+MI) showed increase of DON while reduction of ZEN compared to treatment without MI. Due to fast growth of RO and MI, the co-culture of TR+MI and TR+RO after 3 days of fermentation improved total AA by 32 % and 28 %, respectively, higher than monoculture of TR (18 % after 3 days). Compared with co-culture, monoculture of TR produced feed with higher in vitro dry matter digestibility (IVDMD) with 22–43 % improvement from control, and higher in vitro digestibility of total AA (IVDAA) with 35–50 % improvement from control, the similar level was also achieved by co-culture of TR+RO after 6 days of fermentation. Therefore, co-culture of TR+RO and monoculture of TR in fungal bioprocessing could provide a feasible solution to improve the overall feeding value of the cDDGS.

Tolerance of Rare-Earth Elements in Extremophile Fungus Umbelopsis isabellina from Polar Loparite Ore Tailings in Northwestern Russia

In this study, extremophile fungal species isolated from pure loparite-containing sands and their tolerance/resistance to the lanthanides Ce and Nd were investigated. The loparite-containing sands were collected at the tailing dumps of an enterprise developing a unique polar deposit of niobium, tantalum and rare-earth elements (REEs) of the cerium group: the Lovozersky Mining and Processing Plant (MPP), located in the center of the Kola Peninsula (northwestern Russia). From the 15 fungal species found at the site, one of the most dominant isolates was identified by molecular analysis as the zygomycete fungus Umbelopsis isabellina (GenBank accession no. OQ165236). Fungal tolerance/resistance was evaluated using different concentrations of CeCl3 and NdCl3. Umbelopsis isabellina exhibited a higher degree of tolerance/resistance to cerium and neodymium than did the other dominant isolates (Aspergillus niveoglaucus, Geomyces vinaceus and Penicillium simplicissimum). The fungus began to be inhibited only after being exposed to 100 mg L-1 of NdCl3. The toxic effects of Ce were not observed in fungus growth until it was subjected to 500 mg∙L-1 of CeCl3. Moreover, only U. isabellina started to grow after extreme treatment with 1000 mg∙L-1 of CeCl3 one month after inoculation. This work indicates, for the first time, the potential of Umbelopsis isabellina to remove REEs from the loparite ore tailings, making it a suitable candidate for the development of bioleaching methods.

Divergent Evolution of Early Terrestrial Fungi Reveals the Evolution of Mucormycosis Pathogenicity Factors.

Fungi have evolved over millions of years and their species diversity is predicted to be the second largest on the earth. Fungi have cross-kingdom interactions with many organisms that have mutually shaped their evolutionary trajectories. Zygomycete fungi hold a pivotal position in the fungal tree of life and provide important perspectives on the early evolution of fungi from aquatic to terrestrial environments. Phylogenomic analyses have found that zygomycete fungi diversified into two separate clades, the Mucoromycota which are frequently associated with plants and Zoopagomycota that are commonly animal-associated fungi. Genetic elements that contributed to the fitness and divergence of these lineages may have been shaped by the varied interactions these fungi have had with plants, animals, bacteria, and other microbes. To investigate this, we performed comparative genomic analyses of the two clades of zygomycetes in the context of Kingdom Fungi, benefiting from our generation of a new collection of zygomycete genomes, including nine produced for this study. We identified lineage-specific genomic content that may contribute to the disparate biology observed in these zygomycetes. Our findings include the discovery of undescribed diversity in CotH, a Mucormycosis pathogenicity factor, which was found in a broad set of zygomycetes. Reconciliation analysis identified multiple duplication events and an expansion of CotH copies throughout the Mucoromycotina, Mortierellomycotina, Neocallimastigomycota, and Basidiobolus lineages. A kingdom-level phylogenomic analysis also identified new evolutionary relationships within the subphyla of Mucoromycota and Zoopagomycota, including supporting the sister-clade relationship between Glomeromycotina and Mortierellomycotina and the placement of Basidiobolus as sister to other Zoopagomycota lineages.

Mucormycosis of Forearm Masquerading as Soft Tissue Sarcoma

Mucormycosis is a rare invasive and life-threatening fungal infection usually affecting immunocompromised individuals caused by zygomycetes fungi. Incidence of mucormycosis in immunocompetent individuals is quite rare and is usually attributed to minor trauma. Most common clinical presentations include rhinocerebral and pulmonary forms followed by cutaneous form of mucormycosis where it usually presents as indurated erythematous plaques, targetoid and purpuric lesions. We present a case of mucormycosis in a 34-year-old immunocompetent male presenting with forearm masquerading as soft tissue sarcoma for its rarity in site and clinical presentation.

P289 A teenage with Pythium keratitis—a case report

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM Introduction Pythium is an oomycete found in an aquatic environment and is considered to be a plant pathogen. However, it is able to cause ocular and systemic infections in humans and animals. Keratitis or corneal ulceration caused by Pythium species closely resembles fungal keratitis and is known as a pseudo-fungal infection. It is associated with high ocular morbidity owing to the difficulty in diagnosis and treatment.Case ReportA 13-year-old adolescent from rural Sri Lanka presented with pain and tearing from left eye for 3 weeks’ duration. He was previously healthy and had no history of trauma to the eye. He had bathed in a lake recently. On examination, visual acuity was only perception of light and there was a corneal ulcer in the left eye. Since the ulcer had a poor response to medical therapy, penetrating keratoplasty was performed twice with failure of the graft.Direct smear of both corneal buttons revealed broad aseptate filaments with occasional branches at right angles that mimic fungi of zygomycetes. Culture on Sabouraud Dextrose Agar yielded expanding white submerged colonies. In the tease mount, hyaline, occasionally septate broad fungal filaments were visible. Therefore, the organism was identified as Pythium species in the Mycology Reference Laboratory.Repeated intracameral and intrastromal antifungal and topical and systemic antibiotic treatment resulted in a quiet, vascularized eye with the retained perception of light.Discussion Pythium keratitis carries a significant challenge in laboratory diagnosis due to its mycelial-like appearance. Nested PCR has higher accuracy than standard culture identification. Not being a true fungus, Pythium lacks ergosterol. Therefore, it is usually resistant to many commonly used antifungal agents which target ergosterol. High degree of suspicion is important for accurate identification in the laboratory and the clinicians should be informed for early, aggressive surgical intervention along with antimicrobial therapy in order to achieve a satisfactory outcome.

P288 Rhinocerebral mucormycosis due to Saksenaea vasiformis in a Sri Lankan patient: A rare fungal infection

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PMBackground: Rhizopus, Rhizomucor, and Mucor species are renowned agents causing rhinocerebral mucormycosis; a disease with a high mortality rate. Saksenaea vasiformis is extremely rarely observed in this clinical entity. Minimal sporulation in common laboratory media might be responsible for the under-reporting of this organism. Here we present an uncommon case of rhinocerebral mucormycosis due to S. vasiformis. in a Sri Lankan patient with diabetes mellitus.Case History: A 66-year-old female with diabetes mellitus was admitted with frontal headache, right-sided nasal block, anosmia, and right-side facial swelling. Examination revealed facial edema, maxillary sinus tenderness, and a white patch over the hard palate. Thick pus in postnasal space, growth in posterior tongue base, inflamed palate, and oropharynx, were revealed by rigid nasal endoscopy. She developed ophthalmoplegia along with right V and XII cranial nerve palsies irrespective of antibacterial therapy. Right side pansinusitis was observed in non-contrast computed tomography. She was subjected to right-side full house functional endoscopic sinus surgery with right orbital and optic nerve decompression. Irregular wide, ribbon-like, non-septate hyphae suggestive of Zygomycete fungi were observed in the direct microscopy of a deep surgical tissue sample and started with intravenous amphotericin B. After 5 days of incubation, the culture grew a zygomycetes-like mold with a lack of sporulation on Sabouraud dextrose agar, potato dextrose agar, and slide culture. However, the floating agar technique succeeded in producing flasks-shaped sporangium on a short sporangiophore with brown pigmented rhizoids after 14 days of incubation. The phenotypic features were suggestive of S. vasiformis. She was subjected to repeated debridement surgeries and treatment with amphotericin B was continued. She was clinically improving however, refractory hypokalemia along with hypernatremia interrupted her antifungal therapy resulting return of severe symptoms. Contrast-enhanced computed tomography of the brain revealed multiple micro-abscesses in the right temporal lobe. She clinically deteriorated and succumbed to the illness.Discussion: Saksenaea vasiformis, a member of Mucorales, is largely bounded to cutaneous and subcutaneous infections. Rhino-orbital-cerebral infection is relatively uncommon and most of the reported cases were fatal irrespective of optimal therapy. Prompt diagnosis through fungal investigations of deep biopsy is mandatory. The delayed identification of this organism is attributed to its nature of poor sporulation on routine media. Special culture techniques and nutritionally deprived media enhance sporulation. Saksenaea vasiformis is usually sensitive to amphotericin B, high minimum inhibitory concentration has been reported though.ConclusionSpecific culture techniques should be used to induce sporulation if non-sporulating mucormycetes are encountered. Rhinocerebral mucormycosis is associated with a high degree of mortality even with effective antifungal therapy.

Investigating the Effect of Alcohol Dehydrogenase Gene Knockout on Lipid Accumulation in Mucor circinelloides WJ11

Mucor circinelloides is an oleaginous, dimorphic zygomycete fungus species that produces appreciable levels of ethanol when grown under aerobic conditions in the presence of high glucose, indicating the fungus is a Crabtree-positive microorganism. Engineering efforts to redirect carbon flux from ethanol to lipid biosynthesis may shed light on the critical role of ethanol biosynthesis during aerobic fermentation in M. circinelloides. Therefore, in this study, the alcohol dehydrogenase gene (ADH1) of M. circinelloides WJ11 was deleted, and its effects on growth, lipid production, and fatty acid content were analyzed. Our results showed that knocking out of adh1∆ reduced the ethanol concentration by 85–90% in fermented broth, indicating that this gene is the major source of ethanol production. Parallel to these findings, the lipid and fatty acid content of the mutant was decreased, while less change in the growth of WJ11 was observed. Furthermore, a fermentation study showed the lipid and fatty acid content was restored in the mutant strain when the fermentation media was supplemented with 0.5% external ethanol, indicating the importance of alcohol dehydrogenase and its product on growth and lipid biosynthesis in M. circinelloides. To our knowledge, this is the first study to show a link between alcohol dehydrogenase and lipid production in M. circinelloides.

Evolution of zygomycete secretomes and the origins of terrestrial fungal ecologies

SummaryFungi survive in diverse ecological niches by secreting proteins and other molecules into the environment to acquire food and interact with various biotic and abiotic stressors. Fungal secretome content is, therefore, believed to be tightly linked to fungal ecologies. We sampled 132 genomes from the early-diverging terrestrial fungal lineage zygomycetes (Mucoromycota and Zoopagomycota) and characterized their secretome composition. Our analyses revealed that phylogeny played an important role in shaping the secretome composition of zygomycete fungi with trophic mode contributing a smaller amount. Reconstruction of the evolution of secreted digestive enzymes revealed lineage-specific expansions, indicating that Mucoromycota and Zoopagomycota followed different trajectories early in their evolutionary history. We identified the presence of multiple pathogenicity-related proteins in the lineages known as saprotrophs, suggesting that either the ecologies of these fungi are incompletely known, and/or that these pathogenicity-related proteins have important functions associated with saprotrophic ecologies, both of which invite further investigation.

Fermentative production, characterization and antimicrobial activity of chitosan from some Zygomycetes fungi

Fermentative production, characterization and antimicrobial activity of chitosan from some Zygomycetes fungi

Latent Functions and Applications of Cytochrome P450 Monooxygenases from Thamnidium elegans: A Novel Biocatalyst for 14α-Hydroxylation of Testosterone.

Cytochrome P450 monooxygenases (P450s) are ubiquitous enzymes with high availability and diversity in nature. Fungi provide a diverse and complex array of P450s, and these enzymes play essential roles in various secondary metabolic processes. Besides the physiological impacts of P450s on fungal life, their versatile functions are attractive for use in advanced applications of the biotechnology sector. Herein, we report gene identification and functional characterization of P450s from the zygomycetous fungus Thamnidium elegans (TeCYPs). We identified 48 TeCYP genes, including two putative pseudogenes, from the whole-genome sequence of T. elegans. Furthermore, we constructed a functional library of TeCYPs and heterologously expressed 46 TeCYPs in Saccharomyces cerevisiae. Recombinants of S. cerevisiae were then used as whole-cell biocatalysts for bioconversion of various compounds. Catalytic potentials of various TeCYPs were demonstrated through a functionomic survey to convert a series of compounds, including steroidal substrates. Notably, CYP5312A4 was found to be highly active against testosterone. Based on nuclear magnetic resonance analysis, enzymatic conversion of testosterone to 14α-hydroxytestosterone by CYP5312A4 was demonstrated. This is the first report to identify a novel fungal P450 that catalyzes the 14α-hydroxylation of testosterone. In addition, we explored the latent potentials of TeCYPs using various substrates. This study provides a platform to further study the potential use of TeCYPs as catalysts in pharmaceutical and agricultural industries and biotechnology.

Turning food waste to antibacterial and biocompatible fungal chitin/chitosan monofilaments

Here, cell wall of a zygomycete fungus, Rhizopus delemar, grown on bread waste was wet spun into monofilaments. Using the whole cell wall material omits the common chitosan isolation and purification steps and leads to higher material utilization. The fungal cell wall contained 36.9% and 19.7% chitosan and chitin, respectively. Solid state NMR of the fungal cell wall material confirmed the presence of chitosan, chitin, and other carbohydrates. Hydrogels were prepared by ultrafine grinding of the cell wall, followed by addition of lactic acid to protonate the amino groups of chitosan, and subsequently wet spun into monofilaments. The monofilament inhibited the growth of Bacillus megaterium (Gram+ bacterium) and Escherichia coli (Gram- bacterium) significantly (92.2% and 99.7% respectively). Cytotoxicity was evaluated using an in vitro assay with human dermal fibroblasts, indicating no toxic inducement from exposure of the monofilaments. The antimicrobial and biocompatible fungal monofilaments, open new avenues for sustainable biomedical textiles from abundant food waste.

Antimicrobial activities of microbial essential fatty acid against foodborne pathogenic bacteria.

Background and Objectives:Human health and development have been related to dietary intake of essential fatty acids (omega 3, 6 and 9) and important for brain development, immune system function, and blood pressure regulation. Microbial essential oils are more natural and safer alternatives to synthetic preservatives. These oils have been demonstrated to have antibacterial activity within food systems and may be ideal additives to food formulations. Zygomycete fungi are well-known as good candidate for production of essential oils.Materials and Methods:Essential oils of fungi Mucor rouxii, Mucor circinelloides and Cuninghamella echinulata were extracted and fatty acids were analyzed by GC, for the first, antimicrobial activity of the fungi essential oils against foodborne pathogenic bacteria E. coli, S. aureus, B. cereus, B. subtilis, and S. enterica was examined by disc diffusion and well diffusion methods and the minimal inhibitory concentrations (MIC) of oils were determined by microtiter plate.Results:The fungi oils were exhibited the stron g antibacterial effect against Gram-positive bacteria, B. cereus, S. aureus and B. subtilis higher than Gram-negative and commercial oleic acid and linoleic acid. The MIC of the fungi oil extracts was 0.25 mg/ml for B. cereus and B. subtilis and 0.5 mg/ml about S. aureus. This research demonstrated microbial essential oils may be suitable for their antimicrobial properties in food.Conclusion:Microbial essential oil with good antibacterial activity could also be used in selected cases like foodborne disease.

Review on Antifungal Agents

The four main classes of antifungal drugs are the polyenes, Azoles, allylamines and echinocandins. Clinically useful “older” agents include topical azole Formulations (for superficial yeast and dermatophyte Infections), first-generation triazoles (fluconazole and Itraconazole, for a range of superficial and invasive fungal Infections), amphotericin B formulations (for a broad range of Invasive fungal infections) and terbinafine (for dermatophyte Infections). Clinically important “newer” agents include members of the Echinocandin class (eg, caspofungin) and second-generation Triazoles (eg, voriconazole and posaconazole). Voriconazole and posaconazole have broad-spectrum activity Against yeasts and moulds, including Aspergillus species. Posaconazole is the only azole drug with activity against Zygomycete fungi. Caspofungin and the other echinocandins are effective in Treating Candida and Aspergillus infections. The azoles are relatively safe, but clinicians should be aware Of drug–drug interactions and adverse effects, including Visual disturbances (with voriconazole), elevations in liver Transaminase levels, and skin rashes. Caspofungin has Minimal adverse effects. Combination antifungal therapy may be appropriate in Selected patients with invasive fungal infections, but is Empiric and driven by individual physician practice. Clinical needs for novel antifungal agents have altered.

Cryogenian Glacial Habitats as a Plant Terrestrialisation Cradle - The Origin of the Anydrophytes and Zygnematophyceae Split.

For tens of millions of years (Ma), the terrestrial habitats of Snowball Earth during the Cryogenian period (between 720 and 635 Ma before present–Neoproterozoic Era) were possibly dominated by global snow and ice cover up to the equatorial sublimative desert. The most recent time-calibrated phylogenies calibrated not only on plants but on a comprehensive set of eukaryotes indicate that within the Streptophyta, multicellular charophytes (Phragmoplastophyta) evolved in the Mesoproterozoic to the early Neoproterozoic. At the same time, Cryogenian is the time of the likely origin of the common ancestor of Zygnematophyceae and Embryophyta and later, also of the Zygnematophyceae–Embryophyta split. This common ancestor is proposed to be called Anydrophyta; here, we use anydrophytes. Based on the combination of published phylogenomic studies and estimated diversification time comparisons, we deem it highly likely that anydrophytes evolved in response to Cryogenian cooling. Also, later in the Cryogenian, secondary simplification of multicellular anydrophytes and loss of flagella resulted in Zygnematophyceae diversification as an adaptation to the extended cold glacial environment. We propose that the Marinoan geochemically documented expansion of first terrestrial flora has been represented not only by Chlorophyta but also by Streptophyta, including the anydrophytes, and later by Zygnematophyceae, thriving on glacial surfaces until today. It is possible that multicellular early Embryophyta survived in less abundant (possibly relatively warmer) refugia, relying more on mineral substrates, allowing the retention of flagella-based sexuality. The loss of flagella and sexual reproduction by conjugation evolved in Zygnematophyceae and zygomycetous fungi during the Cryogenian in a remarkably convergent way. Thus, we support the concept that the important basal cellular adaptations to terrestrial environments were exapted in streptophyte algae for terrestrialization and propose that this was stimulated by the adaptation to glacial habitats dominating the Cryogenian Snowball Earth. Including the glacial lifestyle when considering the rise of land plants increases the parsimony of connecting different ecological, phylogenetic, and physiological puzzles of the journey from aquatic algae to terrestrial floras.

Isolation of oleaginous fungi from natural sources, and fatty acid profile of Cunninghamella echinulata

Oleaginous microorganisms accumulate 20% or more of their dry weight as lipid. In the present study, 247 fungal isolates were obtained from different types of soil, marine water, and food, and screened for their ability to accumulate considerable amounts of lipid. A primary screening with Nile red staining was followed by sulfo-phosphovanillin reaction. The most potent isolate was identified employing both morphological and molecular methods as the Zygomycetous fungus, Cunninghamella echinulata AUMC14396. The lipid of this fungus was extracted, transesterified, and analyzed via GC-MS to elucidate the fatty acid (FA) composition. The fatty acid profile revealed the presence of various fatty acids with different concentrations. Of the total fatty acids, saturated FAs comprised 44.49%, while monounsaturated FAs were 19.91%, and polyunsaturated fatty acids (PUFAs) were 35.61%. The most predominant species was linoleic acid (13.79%) followed by stearic (12.78%) and oleic (12.50%) acids. The polyunsaturated fatty acid, gamma linolenic acid was present in a considerable amount (8.02%). Other important PUFAs were also found in the fatty acid profile such as arachidonic acid, eicosapentaenoic acid, and docosahexaenoic acid. The results of this study were discussed in respect to the important applications in the context of production of pharmaceuticals and nutraceuticals.