Cutaneous Mucormycosis in Buffalos in the Brazilian Amazon Biome.

This is the first description of cutaneous mucormycosis in buffalo in the Brazilian Amazon biome. All buffalo showed apathy, inappetence, weight loss, reluctance to move, and prolonged sternal decubitus. Of the four affected animals, two died 15 and 30 days after the appearance of clinical signs. In the initial phase, the skin lesions were rounded areas with dry central regions, sensitive to palpation, with protruding edges and diameters ranging from 8 cm to 15 cm. These areas of necrosis were isolated or coalescing and present mainly on the limbs and sides. In an advanced stage of the disease, there was detachment of the skin from the necrotic areas with extensive wound formation, which sometimes exposed the subcutaneous tissue. The histopathology of the skin showed a multifocal inflammatory infiltrate composed of intact and degenerated eosinophils surrounded by epithelioid macrophages. At the center of these areas was a focally extensive area of epidermal ulceration characterized by intact and degenerated neutrophils, the necrosis of epithelial cells, and the accumulation of fibrin and erythrocytes. The mycological culture was positive for Rhizopus sp. The diagnosis of cutaneous dermatitis caused by Rhizopus sp. was based on clinical signs, macroscopic and histopathological findings, and the identification of the fungus by mycological and molecular techniques.

Inactivation of isolated fungi on Erythrina velutina Willd. seeds through atmospheric plasma

Abstract This study aimed to evaluate the effect of atmospheric plasma application on the inactivation of fungi on the surface of Erythrina velutina seeds and on isolated fungal colonies. Two experiments were conducted using a completely randomized design. First, plasma was applied to the surface of the seeds using helium gas and atmospheric plasma for 3, 6, and 9 min in addition to the control (untreated seeds), constituting seven treatments with five repetitions each. In the second experiment, Petri dishes containing the inoculum of different fungi were treated with atmospheric air plasma for 3, 6, and 9 min (Air-3, Air-6, and Air-9) and were compared with untreated fungi in Petri dishes without treatment (control), totaling four treatments and five repetitions each. We found that the application of atmospheric air plasma to E. velutina seeds for 9 min had an antimicrobial effect on the fungi Aspergillus niger, Aspergillus flavus, Fusarium sp., Brachysporium sp., and Rhizopus sp. The formation of fungal colonies isolated from E. velutina seeds was also inhibited by 3 min of exposure to atmospheric air plasma, except for A. niger, whose inhibition occurred after 6 min of exposure to atmospheric plasma.

Resumo Este trabalho teve como objetivo avaliar o efeito da aplicação de plasma atmosférico na inativação de fungos na superfície de sementes de Erythrina velutina e em colônias fúngicas isoladas. Dois experimentos foram realizados em delineamento inteiramente casualizado: no primeiro, o plasma foi aplicado na superfície das sementes usando gás hélio e plasma atmosférico por três, seis e nove minutos, além do controle (sementes sem tratamento), constituindo sete tratamentos com cinco repetições cada; no segundo experimento, placas de Petri contendo o inóculo de diferentes fungos foram tratadas com plasma atmosférico por três, seis e nove minutos (Air-3, Air-6 e Air-9) e comparadas com fungos não tratados em placas de Petri sem tratamento (controle), totalizando quatro tratamentos e cinco repetições cada. Descobrimos que a aplicação de plasma atmosférico nas sementes de E. velutina por nove minutos teve efeito antimicrobiano sobre os fungos Aspergillus niger, Aspergillus flavus, Fusarium sp., Brachysporium sp. e Rhizopus sp. A formação de colônias fúngicas isoladas de sementes de E. velutina também foi inibida por três minutos de exposição à aplicação de plasma atmosférico, exceto para A. niger, cuja inibição ocorreu a partir de 6 minutos de exposição à aplicação de plasma atmosférico.

Inactivation of isolated fungi on Erythrina velutina Willd. seeds through atmospheric plasma / Inativação de fungos isolados em sementes de Erythrina velutina Willd. por meio do plasma atmosférico

Abstract This study aimed to evaluate the effect of atmospheric plasma application on the inactivation of fungi on the surface of Erythrina velutina seeds and on isolated fungal colonies. Two experiments were conducted using a completely randomized design. First, plasma was applied to the surface of the seeds using helium gas and atmospheric plasma for 3, 6, and 9 min in addition to the control (untreated seeds), constituting seven treatments with five repetitions each. In the second experiment, Petri dishes containing the inoculum of different fungi were treated with atmospheric air plasma for 3, 6, and 9 min (Air-3, Air-6, and Air-9) and were compared with untreated fungi in Petri dishes without treatment (control), totaling four treatments and five repetitions each. We found that the application of atmospheric air plasma to E. velutina seeds for 9 min had an antimicrobial effect on the fungi Aspergillus niger, Aspergillus flavus, Fusarium sp., Brachysporium sp., and Rhizopus sp. The formation of fungal colonies isolated from E. velutina seeds was also inhibited by 3 min of exposure to atmospheric air plasma, except for A. niger, whose inhibition occurred after 6 min of exposure to atmospheric plasma.

Resumo Este trabalho teve como objetivo avaliar o efeito da aplicação de plasma atmosférico na inativação de fungos na superfície de sementes de Erythrina velutina e em colônias fúngicas isoladas. Dois experimentos foram realizados em delineamento inteiramente casualizado: no primeiro, o plasma foi aplicado na superfície das sementes usando gás hélio e plasma atmosférico por três, seis e nove minutos, além do controle (sementes sem tratamento), constituindo sete tratamentos com cinco repetições cada; no segundo experimento, placas de Petri contendo o inóculo de diferentes fungos foram tratadas com plasma atmosférico por três, seis e nove minutos (Air-3, Air-6 e Air-9) e comparadas com fungos não tratados em placas de Petri sem tratamento (controle), totalizando quatro tratamentos e cinco repetições cada. Descobrimos que a aplicação de plasma atmosférico nas sementes de E. velutina por nove minutos teve efeito antimicrobiano sobre os fungos Aspergillus niger, Aspergillus flavus, Fusarium sp., Brachysporium sp. e Rhizopus sp. A formação de colônias fúngicas isoladas de sementes de E. velutina também foi inibida por três minutos de exposição à aplicação de plasma atmosférico, exceto para A. niger, cuja inibição ocorreu a partir de 6 minutos de exposição à aplicação de plasma atmosférico.

Effect of Celeriac Pulp Maceration by Rhizopus sp. Pectinase on Juice Quality.

Peeled and unpeeled celeriac pulp was macerated with pectinase from Rhizopus sp. at 25 °C for 30 and 60 min. Peeling, enzyme addition, and maceration time significantly affected the quality characteristics of the juice. The juice obtained from peeled celeriac was characterized by higher pressing yield, sucrose content, and antioxidant activity (ABTS*+ and DPPH*). The juice from the unpeeled root had higher extract, fructose, glucose, total polyphenols, antioxidant activity (FRAP), total phenolic acids, and total luteolin content. Applying the enzyme to celeriac pulp had no significant effect on the extract's content, analyzed sugars, and antioxidant activity of the juices (ABTS*+). Adding pectinase to unpeeled celery pulp resulted in a 2-10% increase in pressing efficiency, compared to the control sample held at 25 °C for the same period. Maceration of the enzyme-peeled pulp increased the antioxidant potential of the juice by 22% in the FRAP method. In contrast, in all juices analyzed, unpeeled and peeled roots increased antioxidant activity measured by the DPPH* method by 24-57% and total phenolic acids by 20-57%. The time of holding the pulp at 25 °C was an important factor, and its extension resulted in a decrease in the values of most of the analyzed parameters, with the exception of pressing efficiency and fructose content in all analyzed juice samples. Short-term, 30-min maceration of peeled and unpeeled celery pulp with pectinase from Rhizopus sp. had a significant effect on increasing juice yield, antioxidant activity, and phenolic compound content.

Impact of solid-state saccharification on the flavor of rice-flavor baijiu.

Rice-flavor baijiu is a traditional Chinese liquor. The flavor profile and volatiles presented with or without the solid-state saccharification (SSS) were investigated to reveal the effects of SSS process on the quality of rice-flavor baijiu. The liquor prepared with SSS had a sweet flavor. It contained significantly higher contents of ß-phenylethyl alcohol, ß-phenylethyl acetate, and ethyl lactate with odor active value of >1. The liquor prepared without SSS had a cheese-like flavor. It was confirmed that the cheese-like flavor derived from butanoic acid was only detected in the liquor prepared without SSS. SSS facilitated the biosynthesis of ß-phenylethyl alcohol and ethyl lactate by supplying a large amount of phenylalanine and lactic acid at the initial stage of fermentation, and it prevented contamination. These results indicated that the SSS process contributed to produce the characteristic flavor compounds of rice-flavor baijiu. PRACTICAL APPLICATION: Solid-state saccharification (SSS) process of rice-flavor baijiu contributes not only in brewing, but also in the production of the characteristic flavor compounds and the repression of the off-flavor derived from the contamination. Therefore, SSS is a critical process to control the flavor of rice-flavor baijiu.

Hongo del orden mucorales identificado en los ambientes del área Covid ­ 19 de un hospital de alta complejidad / Mucoral order fungus identified in the environments from Covid-19 area of a high complexity hospital

El orden Mucorales está compuesto por los géneros Mucor, Rhizopus y Rhizomucor, estos hongos son de importancia médica y causan mucormicosis en pacientes con una clínica vulnerable como los diabéticos, con un sistema inmune deficiente, enfermedades oncológicas y hematológicas, especialmente los pacientes con insuficiencia respiratoria por la enfermedad del COVID ­ 19 y que a su vez se le asocian las antes mencionadas, facilitando las invasiones por estos agentes oportunistas sin dejar de mencionar las infecciones bacterianas según estudios realizados de aislamientos ambientales

The order Mucorales is composed of the genera Mucor, Rhizopus and Rhizomucor, these fungi are of medical importance and cause mucormycosis in patients with vulnerable clinical conditions such as diabetics, with a deficient immune system, oncological and hematological diseases, especially patients with respiratory failure due to COVID-19 disease, which in turn are associated with the above mentioned, facilitating invasions by these opportunistic agents, not to mention the bacterial infections according to studies of environmental isolates.

Comparative study of As (V) uptake in aqueous medium by a polymer inclusion membrane-based passive sampling device and two filamentous fungi (Aspergillus niger and Rhizopus sp.).

In this work a polymer inclusion membrane (PIM) is proposed as passive sampler material and compared with two filamentous fungi for As (V) uptake to evaluate its ability as chemical surrogate material for the monitoring of this metalloid in aquatic environments. Results show excellent passive sampling characteristics of the device since a linear uptake profile as a function of time was observed. The correlation coefficients between the PIM passive sampler with Aspergillus niger (r = 0.83) and Rhizopus sp. (r = 0.13) uptake, show that the first species is the best modeled by the PIM, suggesting its potential as a chemical substitute in bioavailability studies.

Microbial Community Structure and Chemical Constituents in Shinkiku, a Fermented Crude Drug Used in Kampo Medicine.

Shinkiku (Massa Medicata Fermentata) is a traditional crude drug used to treat anorexia and dyspepsia of elder patients in east Asia. Shinkiku is generally prepared by the microbial fermentation of wheat and herbs. Shinkiku is also used in Japanese Kampo medicine as a component of (Hangebyakujutsutemmato). However, the quality of shinkiku varies by manufacture because there are no reference standards to control the quality of medicinal shinkiku. Thus, we aim to characterize the quality of various commercially available shinkiku by chemical and microbial analysis. We collected 13 shinkiku products manufactured in China and Korea and investigated the microbial structure and chemical constituents. Amplicon sequence analysis revealed that Aspergillus sp. was common microorganism in shinkiku products. Digestive enzymes (α-amylase, protease, and lipase), organic acids (ferulic acid, citric acid, lactic acid, and acetic acid), and 39 volatile compounds were commonly found in shinkiku products. Although there were some commonalities in shinkiku products, microbial and chemical characteristic considerably differed as per the manufacturer. Aspergillus sp. was predominant in Korean products, and Korean products showed higher enzyme activities than Chinese products. Meanwhile, Bacillus sp. was commonly detected in Chinese shinkiku, and ferulic acid was higher in Chinese products. Principal component analysis based on the GC-MS peak area of the volatiles also clearly distinguished shinkiku products manufactured in China from those in Korea. Chinese products contained higher amounts of benzaldehyde and anethole than Korean ones. Korean products were further separated into two groups: one with relatively higher linalool and terpinen-4-ol and another with higher hexanoic acid and 1-octen-3-ol. Thus, our study revealed the commonality and diversity of commercial shinkiku products, in which the commonalities can possibly be the reference standard for quality control of shinkiku, and the diversity suggested the importance of microbial management to stabilize the quality of shinkiku.

Cr(VI) adsorption from aqueous solution by fungal bioremediation based using Rhizopus sp.

The highly toxic species of Chromium in its hexavalent state is an important hazard to the flora and fauna, causing a rupture in balance especially in aquatic environments. The removal of Cr(VI) ions from aqueous solutions using fungal biomass of Rhizopus sp. was investigated under batch experiments. The biomass was produced and treated with NaCl to compare pre-treated and untreated biosorbents capacity. Adsorption of Cr(VI) was investigated with a 23 experimental design to determine the best operational parameters including pH [2.0-4.0], temperature [20-40 °C] and agitation [50-150 rpm]. Maximum Cr(VI) uptake (99%) indicated that pH 2.0 is the optimal for Cr(VI) removal. Linear and non-linear kinetic models were evaluated. The best fitting for linear kinetics was the pseudo-second order linear equation and the Elovich model in its non-linear form, suggesting chemisorption as the controlling step of adsorption. Results followed Langmuir isotherm equation, the qm was 9.95 (mg·g-1) for Rhizopus sp. + NaCl. Thermodynamic parameters were calculated using the adsorption equilibrium constant obtained from Langmuir isotherm and indicated that the adsorption process was spontaneous and endothermic. The surface characteristics of the biomass were analyzed by Fourier transform infrared (FTIR) spectra; the analysis showed the involvement of amino groups in the bonding with Cr(VI). SEM and EDX analysis confirmed the presence of Cr in the biomass after adsorption. The results of these experiments may be utilized for modeling, simulation, and scale-up processes in the future.

Improvement of the Culture Medium for the Dichlorvos-Ammonia (DV-AM) Method to Selectively Detect Aflatoxigenic Fungi in Soil.

The dichlorvos-ammonia (DV-AM) method is a simple but sensitive visual method for detecting aflatoxigenic fungi. Here we sought to develop a selective medium that is appropriate for the growth of aflatoxigenic fungi among soil mycoflora. We examined the effects of different concentrations of carbon sources (sucrose and glucose) and detergents (deoxycholate (DOC), Triton X-100, and Tween 80) on microorganisms in soils, using agar medium supplemented with chloramphenicol. The results demonstrated that 5⁻10% sucrose concentrations and 0.1⁻0.15% DOC concentrations were appropriate for the selective detection of aflatoxigenic fungi in soil. We also identified the optimal constituents of the medium on which the normal rapid growth of Rhizopus sp. was completely inhibited. By using the new medium along with the DV-AM method, we succeeded in the isolation of aflatoxigenic fungi from non-agricultural fields in Fukui city, Japan. The fungi were identified as Aspergillus nomius based on their calmodulin gene sequences. These results indicate that the new medium will be useful in practice for the detection of aflatoxigenic fungi in soil samples including those from non-agricultural environments.

Bioremediation of hexavalent chromium by endophytic fungi; safe and improved production of Lactuca sativa L.

One of the main problems of the industrialized world is the accumulation of chromium (Cr) in soil, which is a serious threat to the crops. Complete removal of Cr from the contaminated soils poses a great challenge. However, this issue can be minimised by using plant growth promoting microbes as a bioremediation tool. In the present study, healthy plants established near the University campus in Mardan were selected for the isolation of Cr resistant endophytes. From the designated plants, 114 species of endophytic fungi were isolated. Among the 114 isolated strains, 4 strains have induced resistance in L. sativa against Cr. The strains were identified as Aspergillus fumigatus, Rhizopus sp., Penicillium radicum and Fusarium proliferatum based on ITS region (18 S rDNA) homology. The isolates have removed Cr from soil and culture media as well as bio-transformed it from highly toxic hexavalent to least toxic trivalent form, thus helped the Cr stressed L. sativa to restore its normal growth. The Rhizopus Sp. CUC23 has mainly accrued Cr and detoxified intracellularly, whereas A. fumigatus ML43 and P. radicum PL17 has detoxified up to 95% of Cr extracellularly. From the results, it is concluded that the selected endophytic strains might be used as biofertilizer for healthy and safe crop production in Cr contaminated soils.

Optimization of thermostable organic solvent-tolerant lipase production by thermotolerant Rhizopus sp. using solid-state fermentation of palm kernel cake.

This study enhanced the production of thermostable organic solvent-tolerant (TS-OST) lipase by locally isolated thermotolerant Rhizopus sp. strain using solid-state fermentation (SSF) of palm kernel cake (PKC). The optimum conditions were achieved using a series of statistical approaches. The cultivation parameters, which include fermentation time, moisture content, temperature, pH, inoculum size, various carbon and nitrogen sources, as well as other supplements, were initially screened by the definitive screening design, and one-factor-at-a-time using PKC as the basal medium. Three significant factors (olive oil concentration, pH, and inoculum size) were further optimized using face-centred central composite design. The results indicated a successful and significant improvement of lipase activity by almost two-fold compared to the initial screening production. The findings showed that the optimal conditions were 2% (v/w) inoculum size, 2% (v/w) olive oil, 0.6% (w/w) peptone, 2% (v/w) ethanol, 70% moisture content at initial pH 10.0 and 45 °C within 72 h of fermentation. Process optimization resulted in maximum lipase activity of 58.63 U/gram dry solids (gds). The analysis of variance showed that the statistical model was significant (p value <0.0001) and reliable with a high value of R2 (0.98) and adjusted R2 (0.96). This indicates a better correlation between the actual and predicted responses of lipase production. By considering this study, the low-cost PKC through SSF appears to be promising in the utilization of agro-industrial waste for TS-OST lipase production. This is because satisfactory enzyme activity could be attained that promises industrial applications.

Quantification of polyphenols and flavonoid content and evaluation of anti-inflammatory and antimicrobial activities of Stenocereus stellatus extracts.

The hexanic, ethyl acetate and methanolic extracts from branches of Stenocereus stellatus were tested in both the 12-O-tetradecanoylphorbol-13-acetate (TPA) - induced ear oedema model and antimicrobial activity assay. The % of oedema inhibition, the Minimum Inhibitory Concentration (MIC), as well as the polyphenolic and flavonoid content were determined. Also, extracts were analysed by gas chromatography-mass spectrometry (GC-MS). In TPA model, the three extracts showed moderate oedema inhibition. In the antimicrobial activity assay, methanolic extract shows better MIC against all strains. The lowest MICs were for Candida albicans (31 µg/mL) and Rhizopus sp. (15 µg/mL). Also, 50.78 mg eq. of gallic acid/g extract of polyphenol and 115.12 mg eq. of catequine/g extract of flavonoids content were founded in ethyl acetate extract. In the chromatographic analysis, ß-sitosterol, ß-amyrine, betulin and some other molecules were identified. The results show that S. stellatus possess antimicrobial activities against some fungus species.

Production of High Ethanol Concentration from Raw Corn Flour Using Rhizopus sp. and Saccharomyces cerevisiae / 微生物学通报

A novel raw-starch-digesting glucoamylase producer,Rhizopus sp.W-08,was used in a novel fermentation system of solid-state followed by submerged,and high enzyme activity of 72 IU/mL was obtained.In the following simultaneous saccharification and fermentation process, Saccharomyces cerevisiae Z-06 directly converted raw corn flour to ethanol with the concentration of 21 % (V/V) at 30℃ after 48h.The conversion efficiency of raw corn flour to ethanol was 94.5 % of the theoretical ethanol yield.