Mucor Ramosissimus Research Articles

Determination of the quantitative and qualitative composition of the bacterial microflora of the digestive tract of horses in parasitic diseases

This article presents the results of studies of the quantitative and qualitative composition of the bacterial microflora of the digestive tract of horses with herd maintenance in Yakutia. The results of a bacteriological study of the intestinal contents conducted in horses infected with intestinal worms showed that the normal composition of the intestinal microflora represented by bacteroids, bifidobacteria, lactobacilli, pathogenic and non-pathogenic E. coli, staphylococci and microscopic fungi changes. As the authors of the article point out, helminths in the process of nutrition violate the integrity of the intestinal wall, blood and lymph vessels, which leads to intestinal bleeding, inflammation and necrosis of intestinal tissues, in addition, migrating through the intestinal wall, helminths are able to carry pathogenic microorganisms that cause infectious diseases (salmonellosus equarum, escherichiosis, adenitis equorum, malleus, etc.). The results of bacteriological studies of the intestinal contents of helminth-infected foals under the age of 6-8 months showed an average growth of bifidobacteria, lactobacilli - 4.3±0.2ґ104 CFU/g, MAFAnM - 1.5±0.1ґ105 CFU/g, non-pathogenic staphylococci - 5.7±0.2ґ104 CFU/g, lactose-negative escherichia - 1.7±0.1ґ103 CFU/g. They noted a high level of pathogenic staphylococci - 1.04±0.1ґ106 CFU/g, the absence of lactose-positive escherichia, a single growth of fungi of the genus Aspergillus, Penicillium and intensive growth of mold fungi of the genus Mucor ramosissimus. When examining the intestinal microflora of adult horses, a low level of bifidobacteria and lactobacilli was found - 7.8±0.3ґ103 CFU/g, MAFAnM was 3.08±0.2ґ105 CFU/g, pathogenic staphylococci - 6.8±0.3ґ104 CFU/g, lactose-negative E. coli - 5.4±0.3ґ102 CFU/g, absence of lactose-positive E. coli, moderate growth of fungi of the genus Mucor ramosissimus and Fusarium dimerum, Candida spp., Penicillium spp. and single growth.

#26: Rhino-Orbital Mucormycosis in Immunocompromised Children: Treatment Outcomes of Three Cases

Abstract Background Mucormycosis is an aggressive opportunistic fungal infection of the family Mucoraceae, including the genera Mucor, Absidia, and Rhizopus. It is the third most common cause of invasive fungal infection, with low incidence, but high mortality (50–90%), and it usually presents in immunocompromised hosts. The fungus spores are ubiquitous in nature and are found in soil, air, and on decaying vegetation. Individuals get infected following inhalation of spores, ingestion, or contamination of wounds. Rhino-orbito-cerebral mucormycosis is the most common form of illness in children. An early diagnosis and a multidisciplinary approach to treatment are necessary to prevent mortality. Methods We present a case series of three immunocompromised children with rhino-orbital mucormycosis in Nacional Medical Center “20 de Noviembre” from 2015 to 2019. We describe time to diagnosis, start of antifungal therapy, surgery involvement, and patient outcomes. Results Patient 1 was a 9-year-old girl with aplastic anemia who developed right palpebral swelling (day 1 of initial symptoms) and was initially diagnosed with preseptal cellulitis. On day 5, a necrotic area appeared in the right inner canthus. Paranasal sinus CT scan showed opacified ethmoidal sinus. Liposomal amphotericin B therapy was started. At day 7, surgical debridement was performed. At day 18, the patient died and the culture of the debrided tissue showed Mucor ramosissimus. Patient 2 was a 15-year-old boy with acute lymphoblastic leukemia who developed a necrotic area in right side of the nose and palate (day 1). Liposomal amphotericin B therapy was initiated. At day 3 and 6, surgery was performed. At day 8, cultures resulted in Rhizopus oryzae, and treatment with caspofungin was added. He had progression of the infection, requiring multiple interventions. Antifungal therapy consisted of 75 days with amphotericin B and 67 days with caspofungin, with resolution. At day 152, he had a event of neutropenia and fever and died of septic shock Patient 3 was a 16-year-old girl with acute lymphoblastic leukemia who developed a necrotic lesion on the palate and right side of the nose (day 1). Direct examination of the lesion showed hyaline, non-septated hyphae. Amphotericin B therapy was initiated and surgery was performed at day 3. By day 7, there was good clinical evolution and resolution the infection. She died at day 12 because of intestinal bleeding and hypovolemic shock. Conclusions The diagnosis and treatment of mucormycosis remains a challenge. Clinical suspicion should be high in patients with risk factors, and early identification and prompt treatment with antifungals and surgical debridement can reduce mortality and improve the prognosis. The poor outcomes of the patients in this case series were mainly due to complications of the underlying disease and not because of mucormycosis. However, in the first case, delayed diagnosis and treatment might have contributed to the unfavorable outcome. The treatment of choice is liposomal amphotericin B. In case 2, a second antifungal therapy included caspofungin due to isolation of R. oryzae, sensitive to echinocandins. Posaconazole can be considered as an alternative option, but it is not available at our institution. Early identification of clinical manifestations and early multidisciplinary treatment with surgical services and antifungal are needed to eliminate the infection. Risk factors must be modified to increase patient survival.

Fungitoxic effect of ethanolic and aqueous leaf extracts of Azadirachta indica A. Juss, Ocimum gratissimum and Benlate in the control of Post harvest Fungal diseases of Solanum melongena Linn (Egg plant).

Fungitoxic properties of ethanolic and aqueous leaf extracts of Azadirachta indica, Ocimum gratissimum and benlate were used in vitro for the control of post-harvest fungal diseases of Solanum melongena, caused by Aspergillus niger, Rhizoctonia solani and Mucor ramosissimus. Pathogenicity test confirmed them as the causal organisms of the fruit rots. The three pathogens were treated with both ethanol and aqueous leaf extracts of the two plants at concentrations ranging from 25 to 100% and benlate at concentrations between 2.5 and 10.0%. Various concentrations were added to prepared Potato Dextrose Agar (PDA) media. The pathogens were inoculated separately into the PDA media and were incubated for eight days. Fungitoxic effects of these extracts on the mycelial growth of the pathogens were significant at P?0.05 for all treatments. The pathogens were completely inhibited at 100% concentration of both extracts and at 10% concentration of benlate. With respect to ethanolic extract; O.gratissimum was more efficient than A. indica but not with aqueous extract.

Neoflavonoids from the heartwood of Dalbergia melanoxylon

Three new neoflavonoids, named (1S,8R,9S)-1,5-dihydroxy-4,12-dimethoxy-8-vinyl-tricyclo[7.3.1.02,7]trideca-2,4,6,11-tetraen-10-one (1), 2,5,2',5'-tetrahydroxy-4-methoxybenzophenone (2) and 2,5,3'-trihydroxy-4-methoxybenzophenone (3), were isolated from the heartwood of Dalbergia melanoxylon. Their structures were established by spectroscopic methods, including UV, IR, HRMS, 1 D and 2 D-NMR. Compounds 1-3 were evaluated for inhibitory activity against three fungal strains Candida albicans, Mucor ramosissimus, Saccharomycopsis fermentans and seven bacterial strains Shigella dysenteriae, Salmonella enteri, Bacillus cereus, Staphylococcus epidermidis, Bacillus sphaericus, Staphylococcus aureus, Escherichia coli using the broth dilution method. However, none of compounds 1–3 showed potential antimicrobial activities in vitro.

A Revised Species Concept for Opportunistic Mucor Species Reveals Species-Specific Antifungal Susceptibility Profiles.

Recently, the species concept of opportunistic Mucor circinelloides and its relatives has been revised, resulting in the recognition of its classical formae as independent species and the description of new species. In this study, we used isolates of all clinically relevant Mucor species and performed susceptibility testing using the EUCAST reference method to identify potential species-specific susceptibility patterns. In vitro susceptibility profiles of 101 mucoralean strains belonging to the genus Mucor (72), the closely related species Cokeromyces recurvatus (3), Rhizopus (12), Lichtheimia (10), and Rhizomucor (4) to six antifungals (amphotericin B, natamycin, terbinafine, isavuconazole, itraconazole, and posaconazole) were determined. The most active drug for all Mucorales was amphotericin B. Antifungal susceptibility profiles of pathogenic Mucor species were specific for isavuconazole, itraconazole, and posaconazole. The species formerly united in M. circinelloides showed clear differences in their antifungal susceptibilities. Cokeromyces recurvatus, Mucor ardhlaengiktus, Mucor lusitanicus (M. circinelloides f. lusitanicus), and Mucor ramosissimus exhibited high MICs to all azoles tested. Mucor indicus presented high MICs for isavuconazole and posaconazole, and Mucor amphibiorum and Mucor irregularis showed high MICs for isavuconazole. MIC values of Mucor spp. for posaconazole, isavuconazole, and itraconazole were high compared to those for Rhizopus and the Lichtheimiaceae (Lichtheimia and Rhizomucor). Molecular identification combined with in vitro susceptibility testing is recommended for Mucor species, especially if azoles are applied in treatment.

CULTURAL PROPERTIES OF FUNGI OF THE FAMILY MUCORACEAE, THE GENUS ASPERGILLUS AS CAUSATIVE AGENTS OF ZOOANTHROPONOSIS

At present, in Ukraine there is a growing prevalence of fungal infections, including aspergillosis and mucormycosis, against HIV / AIDS, tuberculosis, bronchial asthma and cancerous diseases [11]. The literature available presents scanty data on which of the most commonly used nutrient media provide the highest biosynthetic activity for the fungi of the family Mucoraceae, the genus Aspergillus. The aim of this study was to investigate the cultural properties of the studied isolates of fungi on different nutrient media. The following field fungi isolates were studied: Mucor ramosissimus Samutsevitsch, Rhizopus spp., Aspergillus fumigatus Fresenius, Aspergillus niger van Tieghem, Aspergillus flavus Link. The following nutrient media were used: the Plout medium, the Grigoraki medium, the Czapek agar, the Sabouraud agar, the wort-agar, the Van Eterson medium, the Sabouraud-dextrose broth, the honey medium. The concentration of fungi spores per 1 cm3 of inoculum was assessed on the basis of standard methods using a Goryaev's chamber. The evaluation of the intensity of spore formation on different nutrient media was carried out by standard techniques and assessed in CFU/ cm2 [30]. The most intense growth of micromycetes of the Mucor and Aspergillus genera is found on media containing sucrose, dextrose, maltose, glucose, dextrin, glycerol or plant components (the Sabouraud agar, the Czapek agar, the wort agar, the Grigoraki medium, and the Plout medium).

COMPARATIVE BIOLOGICAL EVALUATION OF SIX ENDOPHYTIC FUNGI ISOLATED FROM VINCA ROSEA LEAVES

In this study, a total of six endophytic fungi have been isolated from Vinca rosea (Apocynaceae) leaves growing in Egypt. The isolated fungi were identified morphologically and microscopically up to species to be; Alternaria phragmospora, Aspergillus awamori, Penicillium duclauxii, Penicillium melinii, Nigrospora sphaerica and Mucor ramosissimus. The extracts of the all identified fungi were screened biologically for antileukemic, cytotoxic, antimalarial, antileishmanial, antimicrobial, antioxidant and anti-inflammatory activities as well as for cannabinoid and opioid receptor binding affinities. Out of the six examined fungal extracts four exhibited promising antimalarial activities, four showed modeate antileukemic activities and two of them exhibited cytotoxic activities, three showed antioxidant activities, three exhibited anti-inflammatory activities, and the only one showed an antifungal activity. Alternaria phragmospora was the most active antimalarial agent inhibiting Plasmodium falciparum D6 and W2 clones with IC50 values of 1.9 and 2.1 μg/mL, respectively. Alternaria phragmospora extractwas subjected to liquid-liquid partition using 90% MeOH, hexane then BuOH and H2O. MeOH and BuOH fractions exhibited strong activities against Plasmodium falciparum D6 clone with IC50 values of 1.3 and 28 μg/mL and W2 clone with IC50 values of 1.4 and 28 μg/mL, respectively. These results may be an excellent opportunity to get a new antimalarial drug derived from endophytic fungi.

Isolation and Characterization of Three Zygomycetous Fungi in Korea: Backusella circina, Circinella muscae, and Mucor ramosissimus

While surveying undiscovered fungal taxa in Korea, three rare zygomycetous fungal strains, CNUFC-PTF2-1, CNUFC-TF3-1, and CNUFC-ESAF3-1, were isolated from soil, leaf, and freshwater samples, respectively. The strains were analyzed morphologically as well as phylogenetically based on the internal transcribed spacer region and 28S rDNA sequences. Sequence analysis of the two loci revealed that the isolates, CNUFC-PTF2-1, CNUFC-TF3-1, and CNUFC-ESAF3-1, were identified as Backusella circina, Circinella muscae, and Mucor ramosissimus, respectively. These species have not yet been previously described in Korea.

Isolation, identification and anti-candidal activity of filamentous fungi from Saudi Arabia soil

Ten fungal strains; namely, Penicillium melinii, Petriella setifera, Aspergillus pseudo-niger, Alternaria chlamydospora, Pythium nayoroense, Phoma glomerata, Mucor ramosissimus, Mucor racemosus, Fusarium chlamydosporum and Rhizopus azygosporus were isolated from soil. The extra- and intra-cellular extracts of the fungal strains grown on malt extract and yeast-extract sucrose media were screened for their anticandidal activity against different clinically-isolated Candida species. Most of the fungal extracts showed activity against different Candida species. However, the fungal strains grew on malt extract showed greater activities than those grew on yeast extract sucrose media. The activity of the intracellular was higher than the extracellular metabolites. All fungal extracts (extra and intra) were similar in chemical constituent; they contained carbohydrates and/or glycosides, unsaturated sterols and/or triterpens, tannins and traces of coumarins. Alkaloids, flavonoids, saponins, anthraquinones and cardenolides were no detected. The intra-cellular extracts contained more compounds than the extra-cellular extracts.

Effects of 7-hydroxycalamenene isolated from Croton cajucara essential oil on growth, lipid content and ultrastructural aspects of Rhizopus oryzae.

The leaves and bark of Croton cajucara, a shrub from the Amazon region, have been used in folk medicine to treat diabetes, malaria, and gastrointestinal and liver disorders. The essential oil from the leaves, rich in linalool, presented antileishmanial and antimicrobial activities. A chemotype of this species was found with an essential oil rich in 7-hydroxycalamenene. During our studies of the C. cajucara essential oil, we isolated 7-hydroxycalamenene at > 98 % purity. The minimum inhibitory concentration of 7-hydroxycalamenene against Absidia cylindrospora, Cunninghamella elegans, Mucor circinelloides, Mucor circinelloides f. circinelloides, Mucor mucedo, Mucor plumbeus, Mucor ramosissimus, Rhizopus microsporus, Rhizopus oryzae, and Syncephalastrum racemosum ranged from 19.53 to 2500 µg/mL. The reference drug used, amphotericin B, presented a minimum inhibitory concentration ranging from 0.085 µg/mL to 43.87 µg/mL. 7-Hydroxycalamenene also altered spore differentiation and total lipid content. Ultrastructural analysis by transmission electron microscopy showed significant alterations in the cellular structure of R.oryzae.

Application of a multiplex suspension array for rapid and simultaneous identification of clinically important mold pathogens

We have developed a microsphere-based suspension array (MSA) for the identification of 23 medically important mold pathogens including Aspergillus spp., Fusarium spp., Mucor spp., Rhizopus spp., Rhizomucor pusillus, Penicillium marneffei, Saksenaea vasiformis, Apophysomyces elegans, Lichtheimia corymbifer, and Syncephalastrum racemosum. Twenty-one oligonucleotide probes were designed based on the internal transcribed spacer (ITS2) region for species level identification of molds. Among the 21 probes, 2 probes are shared by more than one species due to low or absence of sequence variability, i.e. Rpam for Rhizopus azygosporus/Rhizopus microsporus and Fumop for Fusarium moniliforme/Fusarium oxysporum/Fusarium pallidoroseum. No cross reactivity was identified except for probes of Mucor racemosus (Murac) which cross react with Mucor hiemalis and Mucor ramosissimus. The sensitivity of MSA is 100 fg–1 ng. The whole procedure including DNA extraction and PCR amplification can be finished within 5 h. The MSA is simple, rapid, specific, high-throughput and capable of multiple-species detection in one reaction tube.

Studies on the Diels-Alder adduct from cyclopentadiene and p-benzoquinone: Biotransformation, enantiomeric excess and absolute configuration

The use of microorganisms or isolated enzymes in synthetic routes has been extensively used by industry and academic research. A great advantage of biotransformation in a synthetic route is highly regio- and enatiosselective control, which can be achieved through microbial catalyzed reaction. Biotransformation is one of the most efficient methods in a production of high purity optical compounds and development of efficient routes for target molecules. Biotransformation by Mucor ramosissimus of enedione 1 gave the keto-alcohol (-)-2 with enantiomeric excess (ee)>99,9% (determined by 1 H NMR with [(+)-Eu(hfc)] 3 ) and their epimer (-)-3 . Determination of the absolute configuration of epimer (-)-3 was accomplished by controlled reaction and measuring of optical rotation (scheme 1). The fungus M. ramosissimus is a promising species to perform desymmetrization of diketones with high enantioselectivity.

Development of new chiral auxiliary from the Diels-Alder adduct endo-tricyclo[6.2.1.02,7] undeca-4 ,9-dien-3 ,6-dione

Stereochemistry control is very important in organic synthesis and it is a basic challenge in asymmetric synthesis. An approach widely used is making use of chiral auxiliary compounds. In this work it is showed the synthesis of highly optically pure rigid polycyclic compounds from the Diels-Alder adduct 1 . Compounds 1 - 3 were prepared as described by Ito et. al. [1]. The alcohol (-)- 3 was obtained by biotransformation of diketone 2 with Mucor ramosissimus, e.e.>99% (60%). The polycyclic halides 4 - 6 were synthesized (60-97%) after treatment of alcohol (-)- 3 with NBS, NIS and, NCS in dichloromethane, respectively. Compound 7 was prepared by reaction with p-toluenesulfonic acid, whilst 8 was prepared by treatment of (-)- 3 with m-chloroperbenzoic acid. The strained polycylic alcohols 9 - 13 will be achieved by reduction of ketones 4 - 8 with NaBH 4 . All compounds were identified by 1 H and 13 C NMR spectroscopy. Due to intrinsic rigidity of the synthesized polycyclic, they could be of use as chiral auxiliary agents. Prochiral substrates can be readily attached by many ways to C-3 or through hydroxyl group found in compounds 8 - 13 . The stereo-dirigent potential of these polycyclic can also be increased by directly or indirectly introduction of many groups at position C-3, or at adjacent positions to carbonyl or hydroxyl at C-3 and C-10.

Antifungal activity of silver nanoparticle-encapsulated β-cyclodextrin against human opportunistic pathogens

Silver nanoparticles were synthesised by reducing silver acetate with a long-chain aliphatic amine. β-Cyclodextrin (CD)-stabilised silver nanoparticles were successfully synthesised and characterised by the UV–vis spectroscopy and scanning electron microscopy analysis. This system was examined for their antifungal activity against opportunistic human pathogens such as Aspergillus fumigatus, Mucor ramosissimus and Chrysosporium species. This study clearly demonstrates that the present system is a powerful antifungal agent against human opportunistic pathogenic fungi.

Two new species ofMucorfrom clinicalsamples

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.

GENOTYPIC ANALYSIS OF MUCOR FROM THE PLATYPUS IN AUSTRALIA

Mucor amphibiorum is the only pathogen known to cause significant morbidity and mortality in the free-living platypus (Ornithorhynchus anatinus) in Tasmania. Infection has also been reported in free-ranging cane toads (Bufo marinus) and green tree frogs (Litoria caerulea) from mainland Australia but has not been confirmed in platypuses from the mainland. To date, there has been little genotyping specifically conducted on M. amphibiorum. A collection of 21 Mucor isolates representing isolates from the platypus, frogs and toads, and environmental samples were obtained for genotypic analysis. Internal transcribed spacer (ITS) region sequencing and GenBank comparison confirmed the identity of most of the isolates. Representative isolates from infected platypuses formed a clade containing the reference isolates of M. amphibiorum from the Centraal Bureau voor Schimmelcultures repository. The M. amphibiorum isolates showed a close sequence identity with Mucor indicus and consisted of two haplotypes, differentiated by single nucleotide polymorphisms within the ITS1 and ITS2 regions. With the exception of isolate 96-4049, all isolates from platypuses were in one haplotype. Multilocus fingerprinting via the use of intersimple sequence repeats polymerase chain reaction identified 19 genotypes. Two major clusters were evident: 1) M. amphibiorum and Mucor racemosus; and 2) Mucor circinelloides, Mucor ramosissimus, and Mucor fragilis. Seven M. amphibiorum isolates from platypuses were present in two subclusters, with isolate 96-4053 appearing genetically distinct from all other isolates. Isolates classified as M. circinelloides by sequence analysis formed a separate subcluster, distinct from other Mucor spp. The combination of sequencing and multilocus fingerprinting has the potential to provide the tools for rapid identification of M. amphibiorum. Data presented on the diversity of the pathogen and further work in linking genetic diversity to functional diversity will provide critical information for its management in Tasmanian river systems.

Biotransformation of a cage-like diels-alder adduct and derivatives by Mucor ramosissimus samutsevitsch

The present study aimed to evaluate the ability for biotransformation of the Diels-Alder adduct tricyclo[6.2.1.0(2,7)]undeca-4,9-dien-3,6-dione (1) and two synthetic derivatives by the saprobe fungus Mucor ramosissimus Samutsevitsch. Products from oxidation, isomerization and, regioselective and enantioselective reduction were achieved.

Isolation of Phosphate-Solubilizing Fungi from Phosphate Mines and Their Effect on Wheat Seedling Growth

Three phosphate-solubilizing fungi, identified as Penicillium expansum, Mucor ramosissimus, and Candida krissii, were isolated from phosphate mines (Hubei, People's Republic of China) and characterized. All the isolates demonstrated diverse levels of phosphate-solubilizing capability in National Botanical Research Institute's phosphate growth medium containing rock phosphate as sole phosphate source. Acidification of culture medium seemed to be the main mechanism for rock phosphate solubilization. Indeed, citric acid, oxalic acid, and gluconic acid were shown to be present in the culture medium inoculated with these isolates. Moreover, the isolates produced acid and alkaline phosphatases in culture medium, which may also be helpful for RP solubilization. A strong negative correlation between content of soluble phosphorus and pH (r = - 0.89; p < 0.01) in culture medium was observed in this study. All the isolates promoted growth, soil available phosphorus, phosphorus, and nitrogen uptake of wheat seedling in field soil containing rock phosphate under pot culture conditions, thus demonstrating the capability of these isolates to convert insoluble form of phosphorus into plant available form from rock phosphate, and therefore hold great potential for development as biofertilizers to enhance soil fertility and promote plant growth.

Biotransformation of a cage-like diels-alder adduct and derivatives by Mucor ramosissimus samutsevitsch

The present study aimed to evaluate the ability for biotransformation of the Diels-Alder adduct tricyclo[6.2.1.02,7]undeca-4,9-dien-3,6-dione (1) and two synthetic derivatives by the saprobe fungus Mucor ramosissimus Samutsevitsch. Products from oxidation, isomerization and, regioselective and enantioselective reduction were achieved.

Spores of Mucor ramosissimus, Mucor plumbeus and Mucor circinelloides and their ability to activate human complement system in vitro

Spores of Mucor ramosissimus, Mucor plumbeus and Mucor circinelloides and their ability to activate human complement system in vitro