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The zygomycete Blakeslea trispora is used commercially as natural source of â-carotene. Trisporic acid (TA) is secreted from the mycelium of B. trispora during mating between heterothallic strains and is considered as a mediator of the regulation of mating processes and an enhancer of carotene biosynthesis. Gas chromatography-mass spectrometry and multivariate analysis were employed to investigate TA-associated intracellular biochemical changes in B. trispora. By principal component analysis, the differential metabolites discriminating the control groups from the TA-treated groups were found, which were also confirmed by the subsequent hierarchical cluster analysis. The results indicate that TA is a global regulator and its main effects at the metabolic level are reflected on the content changes in several fatty acids, carbohydrates, and amino acids. The carbon metabolism and fatty acids synthesis are sensitive to TA addition. Glycerol, glutamine, and ã-aminobutyrate might play important roles in the regulation of TA. Complemented by two-dimensional electrophoresis, the results indicate that the actions of TA at the metabolic level involve multiple metabolic processes, such as glycolysis and the bypass of the classical tricarboxylic acid cycle. These results reveal that the metabolomics strategy is a powerful tool to gain insight into the mechanism of a microorganism's cellular response to signal inducers at the metabolic level. [ABSTRACT FROM AUTHOR]

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Objectives Mucormycetes (formerly known as zygomycetes of the order Mucorales) and hyaline moulds such as those of the genus Fusarium or Paecilomyces are emerging as significant human pathogens. The aim of the study was to determine the in vitro antifungal susceptibility of these fungi to older and newer antifungals and to investigate the antifungal activity of amphotericin B, posaconazole and anidulafungin in dual combinations. Methods Twenty-one clinical isolates of mucormycetes and 16 of rare hyaline moulds were tested. MICs were determined by EUCAST methodology for conidia-forming moulds and Etesting. For antifungal combinations a chequerboard method based on EUCAST methodology was used. Results Against mucormycetes, amphotericin B exhibited the lowest MICs, followed by posaconazole. Ravuconazole was active against eight of the Rhizopus isolates (MIC 1 mg/L). Resistance to amphotericin B (MIC ≥2 mg/L) and posaconazole (MICs ≥4 mg/L) was observed in five and three Rhizopus isolates, respectively. Among Fusarium species variable susceptibility patterns were detected. Amphotericin B exhibited the lowest MICs, followed by voriconazole. Etesting for amphotericin B and posaconazole had excellent agreement with EUCAST methodology (78.6%–100%). Synergy between amphotericin B and anidulafungin was observed against two isolates (one Mucor circinelloides and one Fusarium proliferatum). Synergy or antagonism was not detected in any other combination. Conclusions The study showed that mucormycetes and other rare hyaline moulds exhibit variable susceptibilities to antifungals, and hence antifungal testing is valuable. The fact that the combination of amphotericin B with anidulafungin was found synergistic in some cases merits further investigation. [ABSTRACT FROM PUBLISHER]

Mixed fungal cultures used for making tempe, a fermented soy bean food, were screened for biomass conversion. Thirty-two zygomycetes strains from two tempe cultures were isolated and identified as Rhizopus, Mucor, Rhizomucor, and Absidia species based upon morphology. The dry weight biomass of these strains contained 49% to 63% protein and 10-24% chitosan. The strains with the best growth performance were selected and registered at Culture Collection of Gothenburg University as Rhizomucor CCUG 61146 and Rhizomucor CCUG 61147. These strains were able to grow both aerobically and micro-aerobically. Their ethanol yields were 0.38-0.47, 0.19-0.22, and 0.31-0.38 g/g on glucose, xylose, and a mix sugars consisting of cellobiose, glucose, xylose, arabinose, galactose, and mannose, respectively. The biomass yield of the strains varied between 65 and 140 mg dry weight/g glucose. [ABSTRACT FROM AUTHOR]

Abstract: Cutaneous zygomycosis is a fungal infection caused by zygomycetes that affects the skin. It occurs in uncontrolled diabetic patients and immunosuppressed individuals. It has 2 clinical forms: primary cutaneous zygomycosis and secondary cutaneous zygomycosis. The first is characterized by necrotic lesions and the fungus is usually inoculated by trauma. If diagnosed early, it generally has a good prognosis. Secondary zygomycosis is usually a complication and extension of the rhinocerebral variety that starts as a palpebral fistula and progresses to a necrotic lesion with a poor prognosis. The diagnosis is made by identification of the fungus by direct KOH examination, culture, and biopsy. Treatment for the primary disease is surgical debridement plus amphotericin B. The secondary type is treated with amphotericin B and/or posaconazole. [Copyright &y& Elsevier]

Abstract: An extracellular β-glucosidase from the zygomycete Rhizomucor miehei NRRL 5282 cultivated in a wheat bran-based solid state fermentation system was characterized. The purified enzyme exhibited an optimum temperature of 68–70°C and pH of 5.0. It efficiently hydrolyzed oligosaccharides having β-(1→4) glycosidic linkages and exhibited some β- and α-galactosidase activity. The V max for p-nitrophenyl-β-d-glucopyranoside and cellobiose was 468.2 and 115.5U/mg, respectively, while the Km was 0.12mM for both substrates. The enzyme had transglucosylation and transgalactosylation activities resulting in the formation of glycosides from cellobiose, lactose and ethanol. The enzyme increased the amounts of free phenolic antioxidants in sour cherry pomace indicating that its hydrolyzing activity could potentially be applicable to improve the bioavailability of these compounds. [Copyright &y& Elsevier]

Rhizopus oryzae is a filamentous fungus belonging to the Zygomycetes. It is among others known for its ability to produce the sustainable platform chemicals l-(+)-lactic acid, fumaric acid, and ethanol. During glycolysis, all fermentable carbon sources are metabolized to pyruvate and subsequently distributed over the pathways leading to the formation of these products. These platform chemicals are produced in high yields on a wide range of carbon sources. The yields are in excess of 85 % of the theoretical yield for l-(+)-lactic acid and ethanol and over 65 % for fumaric acid. The study and optimization of the metabolic pathways involved in the production of these compounds requires well-developed metabolic engineering tools and knowledge of the genetic makeup of this organism. This review focuses on the current metabolic engineering techniques available for R. oryzae and their application on the metabolic pathways of the main fermentation products. [ABSTRACT FROM AUTHOR]

The article focuses on a study which aims to determine physiological interaction between Aleuria aurantia lectin (AAL) produced in the mycelia and fruit-bodies of the ascomycete fungus Aleuria aurantia that hampers zygomycete fungus Mucor racemosus growth. It reports that AAL specifically bound to the hyphae of M. racemosus that inhibits its growth. The minimal concentration of AAL required to inhibit and disrupt the growth of fungus is also included.

Abstract: The capability of two zygomycetes strains, Mucor indicus and an isolate from tempeh (Rhizopus sp.), to grow on orange peel hydrolysate and their tolerance to its antimicrobial activity, was investigated. Both fungi, in particular M. indicus, tolerated up to 2% d-limonene in semi-synthetic media during cultivation in shake flasks, under aerobic as well as anaerobic conditions. The tolerance of M. indicus was also tested in a bioreactor, giving rise to varying results in the presence of 2% limonene. Furthermore, both strains were capable of consuming galacturonic acid, the main monomer of pectin, under aerobic conditions when no other carbon source was present. The orange peel hydrolysate was based on 12% (dry w/v) orange peels, containing d-limonene at a concentration of 0.6% (v/v), which no other microorganism has been reported to be able to ferment. However, the hydrolysate was utilised by M. indicus under aerobic conditions, resulting in production of 410 and 400mgethanol/g hexoses and 57 and 75mgfungal biomass/g sugars from cultivations in shake flasks and a bioreactor, respectively. Rhizopus sp., however, was slow to germinate aerobically, and neither of the zygomycetes was able to consistently germinate in orange peel hydrolysate, under anaerobic conditions. The zygomycetes strains used in the present study demonstrated a relatively high resistance to the antimicrobial compounds present in orange peel hydrolysate, and they were capable of producing ethanol and biomass in the presence of limonene, particularly when cultivated with air supply. [Copyright &y& Elsevier]

Abstract: An assemblage of distinctive fungal reproductive units occurs in a fragment of permineralized xylem from the Lower Coal Measures (Carboniferous) of Great Britain. Specimens are spherical to irregular in outline, 70–95μm in diameter, and consist of a central cavity surrounded by an investment of tightly interlaced hyphae. Clusters of specimens may additionally be enveloped in a confluent hyphal meshwork. This complement of features is quite similar to the morphology seen in mantled zygosporangia/zygospores of certain extant zygomycetous fungi. Closely associated with several of the reproductive units are single or paired smaller spherical to elongate structures, which we interpret as apposed gametangia. This discovery represents only the third record of zygomycetous fungi from the Carboniferous. Moreover, the reproductive units resemble certain Carboniferous microfossils termed fungal sporocarps in the genus Mycocarpon, and thus indirectly support the hypothesis that the systematic affinities of these sporocarps may lie with the zygomycetous fungi. [Copyright &y& Elsevier]

The highest lycopene production in mated cultures of Blakeslea trispora was 578 mg/l by adding 42 mg geraniol/l to the medium after 48 h of growth. The control gave 317 mg/l. Adding isopentenyl alcohol at 40 mg/l, mevalonic acid at 17.5 mg/l or dimethyl allyl alcohol at 150 mg, each after 36 h growth, gave lycopene yields 62, 45 and 47%, respectively, higher than the control. [ABSTRACT FROM AUTHOR]

Many case reports suggest that the use of hyperbaric oxygen therapy associated with surgical and medical approaches may contribute to restricting the growth of zygomycetes in patient tissue. The primary aim of this study was to obtain data concerning the in vitro susceptibility of 22 zygomycetes to antifungals such as amphotericin B and posaconazole, and to compare the in vitro development of these fungi in aerobic normobaric versus hyperoxic normobaric and hyperbaric atmosphere. None of the zygomycetes grew after 24-hour and 72-hour incubation in a hyperoxic hyperbaric (2 or 3 ATA) atmosphere. However, when plates were maintained at room temperature in aerobic conditions, colonies were observed from 36-96 h after inoculation, while minimum inhibitory concentration (MIC) values remained the same. This preliminary in vitro study focuses on the in vitro examination of combination therapies to potentiate antifungal activity. Both hyperoxic hyperbaric conditions and a single antifungal agent, as well as combinations of different antifungal drugs were used. Results suggest an impressive in vitro fungistatic activity of the hyperoxic hyperbaric atmosphere, even if the antifungal effect is strictly time-dependent using these incubation conditions. [ABSTRACT FROM AUTHOR]

Studies on the abundance and genera composition of cellulose-decomposing fungi in four differently disturbed peatlands occupied by invasive moss Campylopus introflexus were carried out in autumn, spring and summer seasons. The abundance of fungi under C. introflexus was compared with the abundance of fungal communities under other plants occupying the same habitats (Polytrichum strictum, Eriophorum angustifolium, Calluna vulgaris) and bare peat. The serial dilution plate technique was applied for isolation and enumeration of cellulose-decomposing fungi. The large amount of organic matter as well as acidic reaction of peat creates favourable conditions for the development of fungi at the studied sites. The number of cellulose-decomposing fungi depended on the peatland type, prevailing plants and season. The richness of fungal genera (15) was determined under Campylopus introflexus. Fungal isolates belonging to the Penicillium, Trichoderma, Fusarium genera and the Zygomycetes class prevailed. [ABSTRACT FROM AUTHOR]

Summary Posaconazole is the newest triazole antifungal agent available as an oral suspension with an extended spectrum of activity against Candida species, Aspergillus species, Cryptococcus neoformans, Zygomycetes and endemic fungi. Among posaconazole advantages are the relatively low potential of cross-resistance with other azoles, few drug interactions compared with other azoles and its activity against Zygomycetes. Randomised, double-blind trials have shown that posaconazole is effective for prophylaxis against invasive fungal infections (IFI), especially aspergillosis, in high-risk patients. Results of Phase III clinical trials and case/series reports indicate that posaconazole is effective in treating oesophageal candidiasis, including azole-refractory disease, and other IFI refractory to standard antifungal therapies. To date, posaconazole has appeared to be well tolerated even in long-term courses; it has an excellent safety profile with gastrointestinal disturbances being the most common adverse events reported. The dose of posaconazole is 200 mg three times daily for prophylaxis, 800 mg daily in two or four divided doses for the treatment of IFI and 100 mg daily (200 mg loading dose) for the treatment of oropharyngeal candidiasis. On the basis of early clinical experience, it appears that posaconazole will be a valuable aid in the management of life-threatening fungal infections. [ABSTRACT FROM AUTHOR]

Abstract: The mammalian steroid hormone progesterone actuates a signalling pathway in the zygomycete Rhizopus nigricans which includes heterotrimeric G proteins. To investigate the possibility that the Gβ subunit of these proteins is involved in the signalling, a cDNA library from R. nigricans exposed to progesterone was prepared and a sequence coding for a Gβ subunit was searched for. Using degenerate primers, two sequences, RnGPB1 and RnGPB2, were identified that exhibited a high degree of identity with those for Gβ from other filamentous fungi, but not from yeast. The presence of more than one Gβ subunit is very rare among the fungi, and it has been to date reported only for Rhizopus oryzae. We have shown that progesterone increases the expression of RnGPB1, but has no influence on the expression of RnGPB2. Therefore, our studies imply the involvement of Gβ subunit 1 in the response of R. nigricans to progesterone. Moreover, the Gβ subunit is subjected to endogenous ADP-ribosylation in the presence of NAD, which could be important in some, as yet unknown, cell process. Article from a special issue on steroids and microorganisms. [Copyright &y& Elsevier]

Spent sulphite liquor, the major byproduct from the sulphite pulp production process, was diluted to 50% and used for production of an edible zygomycete Rhizopus sp. The focus was on production, yield, and composition of the fungal biomass composition. The fungus grew well at 20 to 40°C, but 32°C was found to be preferable compared to 20 and 40°C in terms of biomass production and yield (maximum of 0.16 g/g sugars), protein content (0.50-0.60 g/g), alkali-insoluble material (AIM) (ca 0.15 g/g), and glucosamine content (up to 0.30 g/g of AIM). During cultivation in a pilot airlift bioreactor, the yield increased as aeration was raised from 0.15 to 1.0 vvm, indicating a high demand for oxygen. After cultivation at 1.0 vvm for 84 h, high yield and production of biomass (up to 0.34 g/g sugars), protein (0.30-0.50 g/g), lipids (0.02-0.07 g/g), AIM (0.16-0.28 g/g), and glucosamine (0.22-0.32 g/g AIM) were obtained. The fungal biomass produced from spent sulphite liquor is presently being tested as a replacement for fishmeal in feed for fish aquaculture and seems to be a potential source of nutrients and for production of glucosamine. [ABSTRACT FROM AUTHOR]

Mucormycosis is a devastating disease and can occur in patients with a variety of risk factors, the most important of which are immunosuppression, anatomic barrier breakdown, iron overload, and hyperglycemia/ acidosis. Similarly to what occurs with Aspergillus, the host stimulates an innate immune response against the challenging sporangiospores and invading hyphae of Zygomycetes. This article discusses the host defense to different Zygomycetes, its augmentation, and its subsequent impact on the outcome of mucormycosis. [ABSTRACT FROM AUTHOR]

Summary RNA-dependent RNA polymerases (RdRPs) play key roles in the RNA silencing pathway in a number of organisms. They have been involved in the production of double-stranded RNA (dsRNA) molecules that initiate the silencing mechanism as well as in the amplification of the silencing signal. The roles of RdRPs from fungi in these processes are poorly described compared with other eukaryotes. RNA silencing in the zygomycete Mucor circinelloides exhibits uncommon features, such as induction by self-replicative sense transgenes and an amplification process associated with two size classes of antisense small interfering RNAs (siRNAs). To investigate the function of fungal RdRP proteins in initiation and amplification of silencing we have cloned and characterized two different rdrp genes in M. circinelloides. Functional analysis of rdrp- disruption mutants indicates that rdrp-1 is essential for initiation of silencing by sense transgenes by producing antisense RNA transcripts derived from the transgene, but it is not necessary for amplification of the silencing signal, whereas rdrp-2 is required for efficient accumulation of the two different classes of secondary siRNAs regardless the nature of the trigger. Our results provide evidence for a functional diversification of M. circinelloides rdrp genes in different steps of the same RNA silencing pathway. [ABSTRACT FROM AUTHOR]

Zygomycetes such as Cunninghamella elegans seem to be promising biosorbents for pollutants removal from wastewaters because of their particular cell wall characteristics. In this article the effect of ten culture media on C. elegans biomass composition was investigated by means of Fourier transform infra red spectroscopy (FTIR). Biomasses grown on starches from potatoes and cereals were characterised by high amount of chitin and polysaccharides, the glucose gave rise to a biomass rich in acidic polysaccharides and lipids. By contrast, biomasses grown on corn steep liquor were poor in acidic polysaccharides and, when N sources and micronutrients were added, rich in proteins. The lipid content of the biomass generally increased by halving nutrients. Biosorption yields of these biomasses towards four wastewater models were assessed in terms of colour, salts and toxicity reduction. The biomasses rich in proteins and acid polysaccharides were less effective in removing reactive and direct dyes, whereas those rich in cationic polysaccharides showed a higher affinity for these dyes. Both chromatography and FTIR analyses showed that biomasses cultured in halved C and N had the highest affinity for salts. The wastewaters detoxification was quite always achieved, with values often lower that the Italian legal threshold limit. [ABSTRACT FROM AUTHOR]

Several strains of Zygomycetes cultivated on glycerol produced mycelia rich in lipids containing higher amounts of neutral lipids (NL) than glycolipids plus sphingolipids and phospholipids (P), while biosynthesis of P in Mortierella ramanniana, Mucor sp., and Cunninghamella echinulata occurred though NL accumulation process was in progress. Polyunsaturated fatty acids (PUFA) concentration gradually decreased in all lipid fractions of M. ramanniana during growth. In contrast, in C. echinulata concentration of both linoleic and γ-linolenic acids increased with time, especially in P. Taking for granted that the main function of PUFA is associated to their participation in mycelial membranes, we could suppose that biosynthesis of these fatty acids is associated to mycelial growth. However, this is accurate only for some Zygomycetes, e.g., M. ramanniana. On the contrary, PUFA biosynthesis in C. echinulata persists after growth cessation, suggesting that in this species biosynthetic ability is not a strictly growth-associated process. Phosphatidyl-inositol and phosphatidyl-choline were the major P classes in C. echinulata and M. ramanniana, respectively. In M. ramanniana, a decrease of PUFA concentration was noticed even when mycelia were incubated in low temperature (conditions that normally favor PUFA biosynthesis), indicating that PUFA biosynthesis in this fungus is associated to primary metabolism. [ABSTRACT FROM AUTHOR]

Abstract: The metabolite profiles from muscles and livers of Atlantic salmon were investigated using high resolution nuclear magnetic resonance (HR-NMR) spectroscopy of aqueous extracts and magic angle spinning (MAS) NMR spectroscopy of intact tissues. Comparison of the data showed that most small metabolites present in the aqueous extracts were also identified in the Carr–Purcell–Meiboom–Gill (CPMG) MAS NMR spectra of the intact tissues. Not only the total omega-3 fatty acid content, but also the EPA and DHA content, in the muscle and liver tissues could be calculated directly from the diffusion-edited MAS NMR spectra without the need for lipophilic extraction. The effect of replacing a normal fish meal during a period of 3months with a diet containing 20% zygomycete was also investigated by multivariate analysis of the NMR spectra. Principal component analysis (PCA) was used to study the small metabolites distribution in the aqueous extracts of liver and muscles, and revealed that the feed containing zygomycete could have an influence on the metabolites profiles of juvenile Atlantic salmon. [Copyright &y& Elsevier]

A novel biosuperabsorbent protein hydrogel was prepared from protein-rich alcoholic-alkali soluble parts of zygomycete Rhizomucor pusillus biomass. The fungal protein content was 46.8%, and the lipid content was 13.1%. Extraction of protein from this microorganism through the method applied prevents protein decomposition, resulting in maximum yield. After alcoholic-alkaline extraction, the proteins from the biomass were acylated using ethylenediaminetetraacetic dianhydride and subsequently treated with glutaraldehyde as a crosslinker for further experiments. Thermal consistency was investigated by means of two different methods: thermal denaturation via differential scanning calorimetry and thermal decomposition study via thermogravimetric analysis. The swelling behaviour of the crosslinked hydrogel was measured in deionised water, 0.9% NaCl solution and synthetic urine, which were 87.6, 43 and 38.6 g/g water after 24 h, respectively. Moreover, the isoelectric point (p I) of the hydrogel was determined as pH = 8 by studying swelling behaviour at different pHs. In addition, the dependencies of the swelling behaviour with regard to the chemical modification, the ionic strength, the degree of crosslinking, as well as water absorbency with or without load were studied. [ABSTRACT FROM AUTHOR]

Although the fungal order Mortierellales constitutes one of the largest classical groups of Zygomycota, its phylogeny is poorly understood and no modern taxonomic revision is currently available. In the present study, 90 type and reference strains were used to infer a comprehensive phylogeny of Mortierellales from the sequence data of the complete ITS region and the LSU and SSU genes with a special attention to the monophyly of the genus Mortierella. Out of 15 alternative partitioning strategies compared on the basis of Bayes factors, the one with the highest number of partitions was found optimal (with mixture models yielding the best likelihood and tree length values), implying a higher complexity of evolutionary patterns in the ribosomal genes than generally recognized. Modeling the ITS1, 5.8S, and ITS2, loci separately improved model fit significantly as compared to treating all as one and the same partition. Further, within-partition mixture models suggests that not only the SSU, LSU and ITS regions evolve under qualitatively and/or quantitatively different constraints, but that significant heterogeneity can be found within these loci also. The phylogenetic analysis indicated that the genus Mortierella is paraphyletic with respect to the genera Dissophora, Gamsiella and Lobosporangium and the resulting phylogeny contradict previous, morphology-based sectional classification of Mortierella. Based on tree structure and phenotypic traits, we recognize 12 major clades, for which we attempt to summarize phenotypic similarities. M. longicollis is closely related to the outgroup taxon Rhizopus oryzae, suggesting that it belongs to the Mucorales. Our results demonstrate that traits used in previous classifications of the Mortierellales are highly homoplastic and that the Mortierellales is in a need of a reclassification, where new, phylogenetically informative phenotypic traits should be identified, with molecular phylogenies playing a decisive role. [ABSTRACT FROM AUTHOR]

In this article the authors discuss the study which investigates undocumented fungal species. They assess the gap between the described and sequenced dimensions of fungal diversity, focusing on the zygomycete genus. It examines the complexities of nomenclature of mortierelloid fungi. It also demonstrates the need for sequence-based classification of fungal species.

Recent advances in the management of patients with haematological malignancies and transplant recipients have paralleled an increase in the incidence of fungal diseases due to pathogenic genera such as Candida and Aspergillus and the emergence of less common genera including Fusarium and Zygomycetes. Despite availability of new antifungal agents these opportunistic infections have high mortality. Rapid and reliable species identification is essential for antifungal treatment, but detection of the increasing diversity of fungal pathogens by conventional phenotypic methods remains difficult and time-consuming, and the results may sometimes be inconclusive, especially for unusual species. New diagnostic techniques (e.g., 1,3-beta-d-glucan detection) could improve this scenario, although further studies are necessary to confirm their usefulness in clinical practice. [ABSTRACT FROM AUTHOR]

C.M. Durand, C.D. Alonso, A.P. Subhawong, N.P. Kwiatkowski, M. Showel, K.C. Carroll, K.A. Marr. Rapidly progressive cutaneous Rhizopus microsporus infection presenting as Fournier's gangrene in a patient with acute myelogenous leukemia. Transpl Infect Dis 2011: 13: 392-396. All rights reserved Abstract: Members of the genus Rhizopus within the class Zygomycetes can cause devastating opportunistic infections. Cutaneous disease arising from direct inoculation of fungal spores has the potential to disseminate widely. Here, we describe a dramatic case of cutaneous Rhizopus infection involving the penis in a patient with acute myelogenous leukemia. Despite aggressive surgical debridement, systemic antifungal therapy, and donor lymphocyte infusion, the infection was ultimately fatal. This case illustrates the unique diagnostic and therapeutic challenges in the clinical management of cutaneous Rhizopus infection. [ABSTRACT FROM AUTHOR]

Abstract: The zygomycete Mucor circinelloides accumulates β-carotene as the main carotenoid compound. In this study, the applicability of some early genes of the general isoprenoid pathway to improve the carotenoid production in this fungus was examined. The isopentenyl pyrophosphate isomerase gene (ipi) was cloned and used together with the genes encoding farnesyl pyrophosphate synthase (isoA) and geranylgeranyl pyrophosphate synthase (carG) in overexpression studies. Transformation experiments showed that the first bottleneck in the pathway, from the aspect of carotenoid production, is the step controlled by the carG gene, but overexpression of the ipi and isoA genes also contributes to the availability of the precursors. Transformations with these isoprenoid genes in combination with a bacterial β-carotene ketolase gene yielded Mucor strains producing canthaxanthin and echinenone. [Copyright &y& Elsevier]

Abstract: The present work deals with production of ethanol from sweet sorghum bagasse by a zygomycetes fungus Mucor hiemalis. The bagasse was treated with phosphoric acid and sodium hydroxide, with or without ultrasonication, prior to enzymatic hydrolysis by commercial cellulase and β-glucosidase enzymes. The phosphoric acid pretreatment was performed at 50°C for 30min, while the alkali treatment performed with 12% NaOH at 0°C for 3h. The pretreatments resulted in improving the subsequent enzymatic hydrolysis to 79–92% of the theoretical yield. The best hydrolysis performance was obtained after pretreatment by NaOH assisted with ultrasonication. The fungus showed promising results in fermentation of the hydrolyzates. In the best case, the hydrolyzate of NaOH-ultrasound pretreated bagasse followed by 24h fermentation resulted in about 81% of the corresponding theoretical ethanol yield. Furthermore, the highest volumetric ethanol productivity was observed in the hydrolyzates of NaOH pretreated bagasse, especially after ultrasonication in pretreatment stage. [Copyright &y& Elsevier]

Cokeromyces recurvatus is a dimorphic zygomycete with histologic morphology similar to Coccidioides immitis. A 66-year-old man who was status-post bone marrow transplantation for chronic myelogenous leukemia was hospitalized with new onset rash, nausea, and vomiting and subsequently expired. A sputum culture collected on the day of death revealed heavy growth of C. recurvatus 6 days after collection. At autopsy, microscopic examination of the lungs revealed numerous thick-walled, nonbudding spherules ranging in size from 40 to 80 µm. Initial immunohistochemical staining of the formalin-fixed lung tissue was positive for Coccidioides . Additional immunoperoxidase staining revealed the organisms were consistent with a zygomycete fungus, compatible with C. recurvatus infection. Polymerase chain reaction using panfungal primers was attempted on the formalin-fixed tissue but was inconclusive. This case highlights the potential for misdiagnosing Cokeromyces as Coccidioides when the diagnosis is based on histology and immunohistochemical staining. [ABSTRACT FROM PUBLISHER]

The data on visceral mycoses reported in the "Annual of Pathological Autopsy Cases in Japan" were analyzed epidemiologically every four years from 1989 to 2005, and in 2007. The frequency rates of visceral mycoses dropped sharply between 1989 (4.5%) and 1994 (3.2%), but by 2001 had risen again and have remained (4.4-4.6%) generally stable since then. The predominant causative agents were Candida and Aspergillus. Although the rate of candidosis showed a gradual decrease, the rate of aspergillosis showed an increase by degrees. Furthermore, the rate of aspergillosis exceeded that of candidosis in 1994, and the difference in the rates between the two conditions apparently further increased until 2001. After 2005, however no changes in this difference were observed. For complicated infections, the incidence of coinfection with Aspergillus and Candida showed a decreasing, and that with Aspergillus and Zygomycetes showed an increasing tendency. Severe infections with Zygomycetes showed a clear increase from 57.4% in 1989 to 88.9% in 2007. Comparing underlying diseases with mycoses in 1989 and 2007, leukemia (including myelodysplastic syndrome) decreased from 26.1% to 18.8% and bacterial infections (including interstitial pneumonia) increased from 11.1% to 22.1%. By age, the highest frequency rate of mycoses was observed in the range of 60-79 years, and the frequency rate of exogenous fungal infections such as aspergillosis, cryptococcosis, zygomycosis and trichosporonosis showed an increasing trend in the less than one-year old group. [ABSTRACT FROM AUTHOR]

Abstract: To date, among the zygomycete fungi that have been examined, a Spitzenkörper has not been reported. In this paper, the cytoplasmic order of hyphal tip cells of Basidiobolus sp., a zygomycete genus of uncertain phylogeny, has been examined using light microscopy and transmission electron microscopy methods. With phase-contrast light optics, a phase-dark body was observed at the tips of growing hyphae of Basidiobolus sp. The hyphal apex also showed high affinity for FM4-64 labelling resulting in an intense fluorescence signal. The phase-dark inclusion exhibited independent motility within the hyphal apex and its presence and position were correlated to the rate and direction of hyphal growth. The hyphal apex of Basidiobolus sp. did not contain γ-tubulin. Ultrastructural observations revealed a dense cluster of vesicles at the hyphal apex. These results suggest that the growing hypha of Basidiobolus sp. contains a Spitzenkörper, a character generally attributed to members of the ascomycete and basidiomycete fungi and not to zygomycete fungi. [Copyright &y& Elsevier]

Pulmonary mucormycosis is an opportunist fungal infection with high mortality. Posaconazole, an azole antifungal which has a proven efficacy against zygomycetes in vitro, has been shown to be useful as salvage therapy in various studies. We reported herein a lymphoma patient with persistent febrile neutropenia treated for invasive aspergillosis who developed pulmonary mucormycosis subsequently, and shared our experience on his treatment using posaconazole as salvage therapy following amphothericin B, which is thought to enhance survival because of the positive outcome achieved. [ABSTRACT FROM AUTHOR]

Copyright of IBS, Immuno-analyse & Biologie Specialisee is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Abstract: A complete process for the production of bioethanol and fungal biomass from spruce and birch was investigated. The process included milling, pretreatment with N-methylmorpholine-N-oxide (NMMO), washing of the pretreated wood, enzymatic hydrolysis, and cultivation of the zygomycetes fungi Mucor indicus. Investigated factors included wood chip size (0.5–16mm), pretreatment time (1–5h), and scale of the process from bench-scale to 2m high air-lift reactor. Best hydrolysis yields were achieved from wood chips below 2mm after 5h of pretreatment. Ethanol yields (mg/gwood) of 195 and 128 for spruce, and 175 and 136 for birch were achieved from bench-scale and airlift, respectively. Fungal biomass yields (mg/gwood) of 103 and 70 for spruce, and 86 and 66 for birch from bench scale and airlift respectively were simultaneously achieved. NMMO pretreatment and cultivation with M. indicus appear to be a good alternative for ethanol production from birch and spruce. [Copyright &y& Elsevier]

Abstract: Molecular approaches to study the biology of the zygomycete Mucor circinelloides depend mainly on the existence of a polyethylene glycol-based transformation method, which is one of the most efficient in zygomycete fungi. However, the poor reliability and low transformation rates of this method are major obstacles in the molecular study of a number of biological processes. This paper describes an easy and reliable method to transform M. circinelloides protoplasts by electroporation. A high-voltage pulse of 25μF capacitance, 400Ω resistance, and 4kV/cm field strength were seen to be the optimal electrical conditions for delivering DNA into M. circinelloides protoplasts. Under these electrical conditions, successful transformations were carried out with several self-replicative plasmid and strain combinations, producing up to more than 500 transformants per μg DNA. Targeted DNA integration of a transgene (atfA gene of Acinetobacter baylyi) in a particular locus (carRP) was also achieved. This transformation method will considerably facilitate in-depth molecular genetic studies of the biology of this fungus. [Copyright &y& Elsevier]

Skull-base osteomyelitis (SBO) occurs secondary to invasive bacterial and fungal infection. Distinguishing between fungal and bacterial aetiologies of SBO has significant therapeutic implications. An 18-year (1990-2007) retrospective review of patients with SBO presenting to Westmead Hospital was performed. Epidemiological, clinical, laboratory and radiology data were collated. Twenty-one patients (median age 58 years) with SBO were identified: ten (48%) had bacterial and 11 (52%) had fungal SBO. Diabetes mellitus (57%) and chronic otitis externa (33%) were the most frequent co-morbidities; immunosuppression was present in five cases (24%). Cranial nerve deficits occurred in ten (48%) patients. The commonest pathogens were Pseudomonas aeruginosa (50% bacterial SBO) and a zygomycete (55% fungal SBO). Compared to bacterial SBO, fungal SBO was more frequently associated with underlying chronic sinusitis, sinonasal pain, facial/periorbital swelling and nasal stuffiness or discharge and the absence of purulent ear discharge (all p <0.05). Bacterial SBO was more frequently associated with deafness, ear pain or ear discharge (all p <0.05). Median time to presentation was longer in patients with bacterial SBO (26.3 weeks vs. 8.1 weeks, p 0.08). Overall 6-month survival was 88% (14/18 patients). All four deaths occurred in patients with fungal SBO. Immunosuppression was a risk factor for death (p <0.05). Early diagnostic sampling is recommended in patients at increased risk of fungal SBO to enable optimal antimicrobial and surgical management. [ABSTRACT FROM AUTHOR]

Abstract: Fifteen eukaryotic microorganisms were tested for their ability to assimilate biodiesel derived waste glycerol and convert it into value-added metabolic products. For this purpose yeast and Zygomycetes strains were cultivated in nitrogen-limited raw glycerol-based media (initial glycerol concentration 30 g/L). Yeasts tested accumulated restricted lipid quantities (up to ∼22%, wt/wt, in the case of Rhodotorula sp), while differentiations in their fatty acid composition were recorded in relation to the yeast strains employed and the fermentation time. On the contrary, fungi accumulated higher quantities of lipid inside their mycelia (ranging between 18.1 and 42.6%, wt/wt, of dry biomass) that contained in variable amounts the medically important GLA (γ-linolenic acid). Moreover, Yarrowia lipolytica, Pichia membranifaciens and Thamnidium elegans were further studied in media having increased initial glycerol concentrations. In these conditions Y. lipolytica secreted significant amounts of acetic acid (29.2 g/L), as well as mannitol (19.4 g/L) while P. membranifaciens reached 28.4 g/L of biomass at glycerol concentration 90 g/L. T. elegans produced 11.6 g/L of oil, with 71.1%, wt/wt, of fat in biomass, while the maximum concentration of GLA was 371 mg/L. Detailed analysis of T. elegans lipids indicated that the phospholipids fraction was particularly rich in polyunsaturated fatty acids. [Copyright &y& Elsevier]

A new species of Syncephalis (Zoopagales) was isolated from soil in a semiarid area of northeastern Brazil. Syncephalis aggregata is distinguished from the other species of the genus by the production of merosporangiophores in dense tufts and with randomly spaced, irregular swellings and simple merosporangia produced over the upper 50% of a globose to ovoid vesicle. An identification key for the species of Syncephalis found in Brazil is provided. [ABSTRACT FROM AUTHOR]

Background: Burkholderia rhizoxinica is an intracellular symbiont of the phytopathogenic zygomycete Rhizopus microsporus, the causative agent of rice seedling blight. The endosymbiont produces the antimitotic macrolide rhizoxin for its host. It is vertically transmitted within vegetative spores and is essential for spore formation of the fungus. To shed light on the evolution and genetic potential of this model organism, we analysed the whole genome of B. rhizoxinica HKI 0454 - a type strain of endofungal Burkholderia species. Results: The genome consists of a structurally conserved chromosome and two plasmids. Compared to free-living Burkholderia species, the genome is smaller in size and harbors less transcriptional regulator genes. Instead, we observed accumulation of transposons over the genome. Prediction of primary metabolic pathways and transporters suggests that endosymbionts consume host metabolites like citrate, but might deliver some amino acids and cofactors to the host. The rhizoxin biosynthesis gene cluster shows evolutionary traces of horizontal gene transfer. Furthermore, we analysed gene clusters coding for nonribosomal peptide synthetases (NRPS). Notably, B. rhizoxinica lacks common genes which are dedicated to quorum sensing systems, but is equipped with a large number of virulence-related factors and putative type III effectors. Conclusions: B. rhizoxinica is the first endofungal bacterium, whose genome has been sequenced. Here, we present models of evolution, metabolism and tools for host-symbiont interaction of the endofungal bacterium deduced from whole genome analyses. Genome size and structure suggest that B. rhizoxinica is in an early phase of adaptation to the intracellular lifestyle (genome in transition). By analysis of tranporters and metabolic pathways we predict how metabolites might be exchanged between the symbiont and its host. Gene clusters for biosynthesis of secondary metabolites represent novel targets for genomic mining of cryptic natural products. In silico analyses of virulence-associated genes, secreted proteins and effectors might inspire future studies on molecular mechanisms underlying bacterial-fungal interaction. [ABSTRACT FROM AUTHOR]

Two new species in the order Mucorales, Mucor velutinosus and Mucor ellipsoideus, isolated from human clinical specimens in the USA, are described and illustrated. The former species is similar to Mucor ramosissimus, from which it can be differentiated by its ability to grow at 37°C and produce verrucose sporangiospores. Mucor ellipsoideus is also able to grow and sporulate at 37°C like M. indicus, the nearest phylogenetic species in this study, however, the former has narrow ellipsoidal sporangiospores in contrast to the subglobose to ellipsoidal sporangiospores of M. indicus. Analysis of the sequences of the ITS and the D1-D2 regions of the rRNA genes confirmed the novelty of these species. The in vitro antifungal susceptibility of the new species showed that amphotericin B was active against all isolates and posaconazole and itraconazole showed low activity. [ABSTRACT FROM AUTHOR]

Ophiobolins are sesterterpene-type phytotoxins produced by fungi belonging mainly to the genus Bipolaris. In this study, the antifungal effect of ophiobolins A and B on different zygomycetes has been examined. Depending on the zygomycete tested, MIC values of 3.175-50 μg mL were found for ophiobolin A and 25-50 μg mL for ophiobolin B. Ophiobolin A inhibited sporangiospore germination of Mucor circinelloides and caused morphological changes; the fungus formed degenerated, thick or swollen cells with septa. Cytoplasm effusions from the damaged cells were also observed. Fluorescence microscopy after annexin and propidium iodide staining of the treated cells suggested that the drug induced an apoptosis-like cell death process in the fungus. [ABSTRACT FROM AUTHOR]

The objective of this research was to study the diversity of fungi associated with the degradation of para rubber leaf litter. Samples were collected at three periods in 2010: January (during the late rainy season), April (during the dry season) and July (early in the rainy season) in Nakhon Si Thammarat and Songkhla Provinces, both in southern Thailand. Each sample comprised the following three groups; newly fallen leaves, middle stage decaying leaves and old decaying fallen leaves. Moist chamber and dilution pour-plate methods were used. Fungal identification was based on morphological examination under compound and stereo microscopes. A total of 447 species of fungi were identified from the para rubber leaf litter, comprising 405 anamorphic taxa, 38 ascomycetes, two zygomycetes, one oomycete and one basidiomycete. Cladosporium tenuissimum was the dominant species in newly fallen leaves. Kirschsteinothelia sp., Nigrospora sphaerica and Veronaea coprophila were the dominant species in middle stage decaying leaves. In addition to Kirschsteinothelia sp., Lasiodiplodia cf. theobromae, Panchanania jaipurensis, Subulispora procuvata and Veronaea coprophila were the dominant species in old decaying fallen leaves. [ABSTRACT FROM AUTHOR]

The Rhizopus oryzae species complex is a group of zygomycete fungi that are common, cosmopolitan saprotrophs. Some strains are used beneficially for production of Asian fermented foods but they can also act as opportunistic human pathogens. Although R. oryzae reportedly has a heterothallic (+/-) mating system, most strains have not been observed to undergo sexual reproduction and the genetic structure of its mating locus has not been characterized. Here we report on the mating behavior and genetic structure of the mating locus for 54 isolates of the R. oryzae complex. All 54 strains have a mating locus similar in overall organization to Phycomyces blakesleeanus and Mucor circinelloides (Mucoromycotina, Zygomycota). In all of these fungi, the minus (2) allele features the SexM high mobility group (HMG) gene flanked by an RNA helicase gene and a TP transporter gene (TPT). Within the R. oryzae complex, the plus (+) mating allele includes an inserted region that codes for a BTB/POZ domain gene and the SexP HMG gene. Phylogenetic analyses of multiple genes, including the mating loci (HMG, TPT, RNA helicase), ITS1-5.8S-ITS2 rDNA, RPB2, and LDH genes, identified two distinct groups of strains. These correspond to previously described sibling species R. oryzae sensu stricto and R. delemar. Within each species, discordant gene phylogenies among multiple loci suggest an outcrossing population structure. The hypothesis of random-mating is also supported by a 50:50 ratio of plus and minus mating types in both cryptic species. When crossed with tester strains of the opposite mating type, most isolates of R. delemar failed to produce zygospores, while isolates of R. oryzae produced sterile zygospores. In spite of the reluctance of most strains to mate in vitro, the conserved sex locus structure and evidence for outcrossing suggest that a normal sexual cycle occurs in both species. [ABSTRACT FROM AUTHOR]

Invasive fungal disease (IFD) is a life-threatening event in immunocompromised patients, and there is an urgent need for reliable screening methods facilitating rapid and broad detection of pathogenic fungi. We have established a two-reaction real-time PCR assay permitting highly sensitive detection of more than 80 fungal pathogens, covering a large spectrum of moulds, yeasts and Zygomycetes. To assess the clinical potential of the assay, more than 600 consecutive specimens from 125 pediatric patients carrying a high risk of IFD were analyzed. An excellent correlation between PCR positivity and the presence of proven, probable or possible fungal infection according to the European Organization for Research and Treatment of Cancer criteria was demonstrated, as revealed by the sensitivity of the assay of 96% (95% CI: 82-99%). The negative predictive value of the panfungal PCR assay presented was 98% (95% CI: 90-100%), while the specificity and the positive predictive value were 77% (95% CI: 66-85%) and 62% (95% CI: 47-75%), respectively. The results indicate that molecular screening of patients during febrile neutropenic episodes by the assay presented could help prevent unnecessary toxicity resulting from empirical antifungal treatment in individuals who may not be at risk of imminent fungal disease. Our observations raise the possibility that rapid species identification may be required to increase the positive predictive value for impending fungus-related disease. [ABSTRACT FROM AUTHOR]

The article presents a case study of an 11-year-old female, 21-year-old female, and 25-year-old male with cutaneous zygomycosis in Mexico. The risk factors for cutaneous zygomycosis common to the three patients reported have been described as a result of penetrating trauma, and or burns from motor vehicle accidents. In addition, the presence of fungi in soil and on soil-contaminated skin lesions was a consistent pattern in the three cases presented.

Abstract: Light is an environmental signal that modulates many aspects of the biology of zygomycete fungi. Light regulation has been investigated in the zygomycetes Phycomyces blakesleeanus, Mucor circinelloides and Pilobolus crystallinus. Examples of light regulation include the phototropism of the fruiting bodies, the regulation of the development of reproductive structures, and the activation of the biosynthesis of β-carotene. In fungi blue light is perceived by proteins homologous to WC-1, a Neurospora crassa photoreceptor and Zn finger protein that interacts with WC-2 to form a photoresponsive transcription factor complex. Unlike ascomycete and basidiomycete fungi that usually have one wc-1 and one wc-2 gene, several studies have uncovered an unexpected multitude of genes similar to wc-1 and wc-2 in the genomes of several zygomycete fungi. Some of these genes are required for fungal photoresponses, but the function of many of them remains unknown. The presence of multiple wc-1 genes confirms previous suggestions of multiple blue-light photoreceptors in Phycomyces. [ABSTRACT FROM AUTHOR]

Invasive fungal infections remain a serious and life-threatening complication in patients undergoing hematopoietic stem cell transplantation. Since it became clear that lymphocytes, in particular lymphocytes from the T helper 1 (T1) subset, play a critical secondary defense against fungal pathogens, the adoptive transfer of functionally active antifungal T1 cells might be an attractive option to restore adaptive antifungal immune effector mechanisms. Major advances have been made in the generation and characterization of antifungal T cells, which are active against medical important fungi such as Aspergillus spp and Candida spp. However, given the paucity of large homogenous patient populations, major challenges remain in evaluating the clinical usefulness of adoptive antifungal immunotherapy, which should be performed in international collaborative trials. [ABSTRACT FROM AUTHOR]

Objectives: We analysed the in vitro and in vivo effects of posaconazole and amphotericin B against three clinical isolates of zygomycetes: Lichtheimia corymbifera, F1; and Rhizopus oryzae, F5 and F6. [ABSTRACT FROM PUBLISHER]

Trisporic acid, its precursors and derivatives are used within zygomycete fungi as communication signals and sexual regulators, and also influence the production rate of the parent compound, β-carotene. Cultivation parameters during growth and the trisporoid production phase of Blakeslea trispora were studied in two-step shake flask cultures and up-scaled fermentations. Comparison of various fermentation protocols allowed the definition of parameters governing trisporoid production. Highest yields were obtained when the initial growth phase allowed for both rapid growth and fast exhaustion of nitrogen and phosporous sources. Onset of trisporoid production is accompanied by a pH drop in the medium and triggered by nutrient limitation, nitrogen depletion being the most important factor. Supplementation of cultures with carbon at low concentration after onset of trisporoid production led to prolonged growth and higher final product accumulation. B. trispora produces trisporoids in two major series, B and C. During a first peak in trisporic acid accumulation, production of trisporic acid B exceeds that of trisporic acid C, which later accumulates at the expense of the trisporic acid B, indicating a variable regulation of the ratio between these metabolites. These data are valuable for tailoring production systems for enrichment of specific intermediates of this complex signal family. [ABSTRACT FROM AUTHOR]

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