Negative correlation between phospholipase and esterase activity produced by Fusarium isolates

Fusarium species have emerged as one of the more outstanding groups of clinically important filamentous fungi, causing localized and life-threatening invasive infections with high morbidity and mortality. The ability to produce different types of hydrolytic enzymes is thought to be an important virulence mechanism of fungal pathogens and could be associated with the environment of the microorganism. Here, we have measured the production of two distinct lipolytic enzymes, phospholipase and esterase, by sixteen Fusarium isolates recovered from the hospital environment, immunocompromised patients’ blood cultures, foot interdigital space scrapings from immunocompromised patients, and foot interdigital space scrapings from immunocompetent patients (4 isolates each). Fourteen of these 16 isolates were identified asFusarium solani species complex (FSSC) and two were identified as F. oxysporum species complex (FOSC). Some relevant genus characteristics were visualized by light and electron microscopy such as curved and multicelled macroconidia with 3 or 4 septa, microconidia, phialides, and abundant chlamydospores. All Fusarium isolates were able to produce esterase and phospholipase under the experimental conditions. However, a negative correlation was observed between these two enzymes, indicating that a Fusarium isolate with high phospholipase activity has low esterase activity and vice versa. In addition, Fusarium isolated from clinical material produced more phospholipases, while environmental strains produced more esterases. These observations may be correlated with the different types of substrates that these fungi need to degrade during their nutrition processes.

The left lung is preferentially targeted during experimental paracoccidioidomycosis in C57BL/6 mice

Paracoccidioidomycosis (PCM) is a chronic systemic mycosis caused by the inhalation of the thermally dimorphic fungus Paracoccidioides brasiliensis as well as the recently described P. lutzii. Because the primary infection occurs in the lungs, we investigated the differential involvement of the right and left lungs in experimental P. brasiliensis infection. Lungs were collected from C57BL/6 mice at 70 days after intravenous infection with 1×106 yeast cells of a virulent strain of P. brasiliensis (Pb18). The left lung, which in mice is smaller and has fewer lobes than the right lung, yielded increased fungal recovery associated with a predominant interleukin-4 response and diminished synthesis of interferon-γ and nitric oxide compared with the right lung. Our data indicate differential involvement of the right and left lungs during experimental PCM. This knowledge emphasizes the need for an accurate, standardized protocol for tissue collection during studies of experimental P. brasiliensis infection, since experiments using the same lungs favor the collection of comparable data among different mice.

Microbial profile of a kefir sample preparations: grains in natura and lyophilized and fermented suspension

Probiotics are supplementary foods developed by microbial strains that improve animal health beyond basic nutrition. Probiotics are consumed orally, regardless of being considered as normal inhabitants of the intestines, able to survive in enzimatic and biliary secretions. Kefir is a probiotic originated from the old continent, fermented by several bacteria and yeasts, encapsulated in a polyssacharide matrix, and resembles jelly grains. Kefir is also presented as its sourish product both in sugary or milky suspensions containing vitamins, aminoacids, peptides, carbohydrates, ethanol, and volatile compounds. Kefir is known to have a diverse microbial content depending on the country and fermentative substrates, which cause distinct probiotic effects. In this sense, the purpose of this work was to isolate, identify, and quantify the microbial content of a native sugary kefir sample (fermented suspension and lyophilized natural grains). Serial dilutions were plated on Rogosa agar (AR) and De Man, Rogosa and Sharpe (MRS), for Lactobacillus; Brain Heart Infusion (BHI), for total bacteria; Sabouraud-Dextrose-Agar (SDA), for yeasts and filamentous fungi; Thioglycolate Agar (TA), for Streptococcus, Acetobacteria and Leuconostoc; and Coconut Water Agar (CWA), and CWA supplemented with yeast extract (CWAY), for various genera. Genera and species for all strains were identified through biochemical reactions and specific API systems. The microbial profile of kefir was different from other sources of grains despite the presence of similar microorganisms and others which have not been reported yet. The data obtained with the CWA and CWAE media suggest that both substrates are alternative and salutary media for culture of kefir strains.

Influence of the fungi population on the physicochemical and chemical composition of coffee (Coffea arabica L.)

The influence of fungi associated with coffee fruits was verified regarding the chemical and physicochemical composition of Coffea arabica L. raw grains. The fruits were harvested at EPAMIG Experimental farm in Lavras, State of Minas Gerais - making up the different samples here analyzed. After processing and drying, the grains were incubated in wet chamber for fungal exteriorization through the blotter test method and submitted to the following analyses: polyphenoloxidase, total reducing and non-reducing sugars, clorogenic acid, titrable acidity, potassium leaching, electric conductivity and caffeine. The occurrence of the P. variable, P. rugulosum, P. funiculosum, F. equiseti, F. semitectum, A.alutaceus, A. niger and C. cladosporioides fungi in the different samples was detected. From the analysis of the results obtained, it was observed that the presence of the Aspergillus alutaceus fungus reduces the activity of the enzyme polyphenoloxidase and increases the values of potassium leaching, electric conductivity and chlorogenic acid. The incidence of the Cladosporium cladosporioides fungus influenced the average values of potassium leaching and electric conductivity.

Phenolic compounds and antioxidant activity in fermented rice (Oryza sativa) bran

This study investigated the content of total phenolic compounds and antioxidant activity in fermented rice bran in order to evaluate the effect of solid state fermentation on these properties. The process was performed with the fungus Rhizopus oryzae CTT 1217 in tray reactors at 30 °C for 120 hours. Samples of fermented rice bran were collected every 24 hours. Antioxidant property was evaluated by the diphenyl-1-picrylhydrazyl radical scavenging method and through the inhibition of enzymatic oxidation and lipid peroxidation of olive oil. The methanol extract of the biomass obtained at 96 hours of fermentation inactivated 50% of free radical in 15 minutes. The same extract reduced the peroxide value in the olive oil by 57% after 30 days of storage. The aqueous extract of the biomass obtained at 120 hours was the most efficient inhibitor of the darkening reaction catalyzed by peroxidase.

Chemical, biochemical, and microbiological aspects of chitosan quaternary salt as active coating on sliced apples

The biocompatibility of chitosan and chitosan quaternary salt coatings was evaluated for use as edible coatings for sliced apple. Measurement of water loss, color change, and fungal growth appearance were monitored as a function of time. A significant brownish effect was observed on chitosan coated slices, varying greatly from L* = 76.5 and Hue angle = 95.9° (t = 0) to L* = 45.3 and Hue angle = 69.8° (t = 3 days), whilst for TMC coated samples the variation was considerable lower (L* = 74.1; Hue angle = 95.0°) to (L* = 67.0; Hue angle = 83.8°) within the same period. The hydrosoluble derivative N,N,N-trimethylchitosan demonstrated good antifungal activity against P. expansum although highly dependent on the polymer properties such as degree of quaternization. The most efficient formulation was that prepared from derivative having a degree of quaternization of 45%, high solubility, and high viscosity. This formulation restrained fungus spreading up to 30%, while for the control it reached almost 80% of the total assessed surfaces during 7 days of storage.

Chemical composition of tempeh from soybean cultivars specially developed for human consumption

Tempeh is a food obtained by fermentation of soybean grains by the fungus Rizophus oligosporus. It is a traditional Indonesian food that presents benefits for human health protecting against diarrhea and chronic diseases. Tempeh processing includes dehulling, cooking, inoculation, and fermentation. In this study, chemical characteristics of tempeh prepared with soybean cultivars specially developed for human consumption (BRS 216, BRS 232, BRS 257, and BRS 267) were investigated. Soybean grains and tempeh obtained from these cultivars were analyzed for oil, protein, antinutrional factors, and isoflavone content. Cultivar BRS 216 presented the highest protein content in the grains (36.81%) and in tempeh (51.99%). On average, the protein content in tempeh increased 16% in relation to that of soybean grains. Isoflavone content was higher in the grains than in tempeh with significant differences among the cultivars. However, the aglycones content increased about 50% in tempeh (49.00 mg.100 g-1 on average) compared to that of raw material (soybean grains - 21.49 mg.100 g-1, on average). The content of Kunitz trypsin inhibitor (KSTI) reduced 83% in tempeh, on average, as compared to the value found in the grains. Phytic acid content was similar in both tempeh and the grains.

Physicochemical and microbiological quality of raspberries (Rubus idaeus) treated with different doses of gamma irradiation

This study was conducted to evaluate the physicochemical and microbiological characteristics of raspberries exposed to different radiation doses. The fruits were harvested in the city of Campestre, MG, packed in polyethylene bags, and transported to the Federal University of Lavras (UFLA), where they were separated into 4 lots. Irradiation was performed at the Center for Development of Nuclear Technology in Belo Horizonte, MG. The doses used were 0 (control), 0.5, 1.0, and 2.0 kGy. After irradiation, the fruits were transported back to UFLA and stored at 1 ºC and 95% relative humidity (RH) for 12 days. The physicochemical analyses for mass loss, total soluble solids, titratable acidity, pH, total soluble sugars, total soluble pectin, firmness, vitamin C content, total antioxidant activity, and total phenolic, and the microbiological assays (coliform at 35 and 45 ºC, psychrotrophic and filamentous fungi and yeasts) were performed after 0, 3, 6, 9, and 12 days of storage. Lower loss of mass and filamentous fungi and yeast count were observed in the irradiated fruits, and 2 kGy was determined as the most effective dose for microbial control, but this irradiation dose also resulted in increased loss of fruit firmness.

Tempeh flour as a substitute for soybean flour in coconut cookies

The objective of this study was to prepare roasted and lyophilized tempeh flour with soybean cultivar BRS 267 to apply them in the formulation of coconut biscuits. The cookies produced with whole soy flour and mixed flour of soybean and tempeh were evaluated for proximate composition, fatty acid profile, and isoflavone aglycones in order to verify the effects of inoculation with the fungus Rhizopus oligosporus and those of the drying processes of roasting and lyophilization on the chemical characteristics of the final product. Sensory acceptance and purchase intention of the formulated products were also evaluated. The results indicate the maintenance of linolenic acid, which is important in the prevention of coronary diseases, and an increase in the aglycones levels when the tempeh flour was used. Lipids and proteins showed differences, and the sensory analyses demonstrated similarity between the cookies with satisfactory scores for aroma, flavor, texture, and overall acceptability for both samples. when compared to the control. Purchase intent was also positive for the lyophilized and toasted tempeh flours, thus enabling the use of the roasting process as a simple drying method, for processing tempeh and obtaining a flour rich in proteins and aglycones that can be used as a partial substitute for soy flour in cookies and other bakery products.

Huitlacoche yield in some maize varieties in the Mediterranean region of Turkey

Huitlacoche is the Aztecs name given to the smut galls on ears of maize caused by the pathogenic plant fungus Ustilago maydis [(DC) Corda.)]. It is known as maize mushroom, and it has been considered a delicacy and in Mesoamerica. The aim of the present study was to determine the responses of some maize varieties to the growth of the fungus in order to evaluate the prospect production of these smutty ears as a maize mushroom. A 2-year study was conducted in the Mediterranean region of Turkey in 2010 and 2011. Inoculations were performed by injecting inoculum into the ear through the silk channel of plants in plots. Each treatment had control plots. Average ear-gall (huitlacoche) severity and incidence of all the varieties were at the rates of 4.0 and 41.6%, respectively. However, the highest severity of ear-gall (6.5) and incidence (60.6%) were found in Karadeniz Yıldızı flint maize variety; colossal smutty ears were formed in the maize cultivars. This study showed that certain maize cultivars (flint corn and dent corn) can be used efficiently in the production of huitlacoche.

Effect of huitlacoche (Ustilago maydis DC Corda) paste addition on functional, chemical and textural properties of tortilla chips

AbstractThis study analyzed the addition of huitlacoche paste (HP) in baked tortilla chips (TC), evaluating its effects on functional, physicochemical and structural changes during processing. Two blue corn grains were nixtamalized, stone milled, air dried and milled to obtain flour; commercial blue corn flour (TM1) and commercial TC (TM2) were used as controls. Additions of 0, 3, 6 and 9% of HP were formulated; masas were prepared at 55% moisture content (MC), precooked and baked in an industrial machine. TC crispiness was influenced by grain characteristics and percentage of HP. Huitlacoche paste addition caused an increase in total dietary fiber (from 5.27 to 14.54%), total soluble phenolics content (from 17.52 to 37.60 mg GAE/100 g) and antioxidant capacity (from 6.74 to 7.98 μmol TE/g) in TC. Results suggest that tortilla chips added with huitlacoche can be an alternative to prepare this traditional edible fungus and produce healthier snacks, not fried and enriched with bioactive compounds.

Enhanced extraction of phenolic compounds from coffee industry’s residues through solid state fermentation by Penicillium purpurogenum

Abstract The use of agroindustrial residues is an economical solution to industrial biotechnology. Coffee husk and pulp are abounding residues from coffee industry which can be used as substrates in solid state fermentation process, thus allowing a liberation and increase in the phenolic compound content with high added value. By employing statistical design, initial moisture content, pH value in the medium, and the incubation temperature were evaluated, in order to increase the polyphenol content in a process of solid state fermentation by Penicillium purpurogenum. The main phenolic compounds identified through HPLC in fermented coffee residue were chlorogenic acid, caffeic acid, and rutin. Data obtained through HPLC with the radical absorbance capacity assay suggest the fermented coffee husk and pulp extracts potential as a source of phenolic acids and flavonoids. Results showed good perspectives when using P. purpurogenum strain to enhance the liberation of phenolic compounds in coffee residues.

Characterisation of Cladonia rangiferina transcriptome and genome, and the effects of dehydration and rehydration on its gene expression

Lichens are symbiotic organisms, which consist of the fungal partner and the photosynthetic partner, which can be either an alga or a cyanobacterium. In some lichen species the symbiosis is tripartite, where the relationship includes both an alga and a cyanobacterium alongside the primary symbiont, fungus. The lichen symbiosis is an evolutionarily old adaptation to life on land and many extant fungal species have evolved from lichenised ancestors. Lichens inhabit a wide range of habitats and are capable of living in harsh environments and on nutrient poor substrates, such as bare rocks, often enduring frequent cycles of drying and wetting. Most lichen species are desiccation tolerant, and they can survive long periods of dehydration, but can rapidly resume photosynthesis upon rehydration. The molecular mechanisms behind lichen desiccation tolerance are still largely uncharacterised and little information is available for any lichen species at the genomic or transcriptomic level. The emergence of the high-throughput next generation sequencing (NGS) technologies and the subsequent decrease in the cost of sequencing new genomes and transcriptomes has enabled non-model organism research on the whole genome level. In this doctoral work the transcriptome and genome of the grey reindeer lichen, Cladonia rangiferina, were sequenced, de novo assembled and characterised using NGS and traditional expressed sequence tag (EST) technologies. RNA extraction methods were optimised to improve the yield and quality of RNA extracted from lichen tissue. The effects of rehydration and desiccation on C. rangiferina gene expression on whole transcriptome level were studied and the most differentially expressed genes were identified. The secondary metabolites present in C. rangiferina decreased the quality – integrity, optical characteristics and utility for sensitive molecular biological applications – of the extracted RNA requiring an optimised RNA extraction method for isolating sufficient quantities of high-quality RNA from lichen tissue in a time- and cost-efficient manner. The de novo assembly of the transcriptome of C. rangiferina was used to produce a set of contiguous unigene sequences that were used to investigate the biological functions and pathways active in a hydrated lichen thallus. The de novo assembly of the genome yielded an assembly containing mostly genes derived from the fungal partner. The assembly was of sufficient quality, in size similar to other lichen-forming fungal genomes and included most of the core eukaryotic genes. Differences in gene expression were detected in all studied stages of desiccation and rehydration, but the largest changes occurred during the early stages of rehydration. The most differentially expressed genes did not have any annotations, making them potentially lichen-specific genes, but several genes known to participate in environmental stress tolerance in other organisms were also identified as differentially expressed.

Efficiency of methods to detect Sclerotinia sclerotiorum in commercial soybean seed lots

Most soybean pathogens are seed transmitted, deserving emphasis the fungus Sclerotinia sclerotiorum, which has been presenting worrying levels of field incidence in some soybean cropping areas in several Brazilian states. The objective of this study was to verify the efficiency of different methods for detecting S. sclerotiorum on soybean seeds artificially infected in the laboratory and from field production areas with a historical disease incidence. Seed samples of seven different cultivars collected from naturally infested fields, and one seed sample artificially inoculated in the laboratory were used. The following detection methods recommended in the literature were compared: Blotter test at 7 ºC, 14 ºC, and 21 ºC; Rolled Paper; and Neon-S. Results demonstrated that these methods showed no repeatability and had a low sensitivity for detecting the pathogen in seeds from areas with disease incidence. They were effective, however, for its detection on artificially inoculated seeds. In the Blotter test method at 7 ºC, there was a lower incidence of other fungi considered undesirable during seed analysis.