Synthesis and antimicrobial activity testing of quaternary ammonium silane compounds.

With increasing health awareness of the pathogenic effects of disease-causing microorganisms, interest in and use (of medical textiles, disinfectants in medical devices, etc.) of antimicrobial substances have increased in various applications, such as medical textiles and disinfectants (alcohol-based and nonalcoholic), in medical devices There are several concerns with alcohol-based disinfectants, such as surface deformation of medical devices due to high alcohol content and damage to skin tissue caused by lipid and protein denaturation of cell membranes. Quaternary ammonium compounds (quats) were preferred because they have the potential to prepare water-based disinfectants. In this study, novel (3-chloropropyl)triethoxysilane (CPTMO) and (3-chloropropyl)triethoxysilane (CPTEO) based quaternary ammonium silane compounds (silane-quats) were developed using quats with carbon chain lengths of C12, C14, C16 and C18. Titration (ASTM D2074) was used to calculate the yield of the synthesis and the structures of the products were characterised by Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance spectroscopy (13C NMR, 1H NMR) and gas chromatography-mass spectrometry (GC-MS).The in vitro antimicrobial activity of the synthesized samples was evaluated against Gram-positive (Staphylococcus aureus (S. aureus), Enterococcus hirae (E. hirae)) and Gram-negative (Escherichia coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa)) bacteria and fungi (Candida albicans (C. albicans), Aspergillus brasiliensis (A. brasiliensis)) using the minimum inhibitory concentration (MIC) test. According to MIC tests, the silane-quats with the highest antimicrobial effects were dimethylhexadecyl[3-(trimethoxysilyl)propyl]ammonium chloride (SQ3), which had an MIC of < 16 µg/ml (ppm) against E. coli, S. aureus, E. hirae, C. albicans, and A. brasiliensis and 32 µg/ml against P. aeruginosa. The MIC test results also showed antimicrobial activity at least 2 times greater than that of the commercially available disinfectant benzalkonium chloride (BAC). Findings suggest that SQ3 (C16) holds promise as an effective medical disinfectant, presenting a novel approach to combating microbial infections in healthcare settings.

Modeling the growth of Aspergillus brasiliensis affected by a nonthermal plasma.

AIM: The main objective of the study was to develop and validate a model for the growth of Aspergillus brasiliensis on surfaces, specifically on agar culture medium. An additional aim was to determine conditions for complete growth inhibition of this micromycete using two different nonthermal plasma (NTP) sources. METHODS AND RESULTS: The developed model uses two key parameters, namely the growth rate and growth delay, which depend on the cultivation temperature and the amount of inoculum. These parameters well describe the growth of A. brasiliensis and the effect of NTP on it. For complete fungus inactivation, a single 10-minute exposure to a diffuse coplanar surface barrier discharge was sufficient, while a point-to-ring corona discharge required several repeated 10-minute exposures at 24-h intervals. CONCLUSIONS: The article presents a model for simulating the surface growth of A. brasiliensis and evaluates the effectiveness of two NTP sources in deactivating fungi on agar media.

Comparing resistance of bacterial spores and fungal conidia to pulsed light and UVC radiation at a wavelength of 254 nm.

Pulsed light (PL) inactivates microorganisms by UV-rich, high-irradiance and short time pulses (250 µs) of white light with wavelengths from 200 nm to 1100 nm. PL is applied for disinfection of food packaging material and food-contact equipment. Spores of seven Bacillus ssp. strains and one Geobacillus stearothermophilus strain and conidia of filamentous fungi (One strain of Aspergillus brasiliensis, A. carbonarius and Penicillium rubens) were submitted to PL (fluence from 0.23 J/cm2 to 4.0 J/cm2) and UVC (at λ = 254 nm; fluence from 0.01 J/cm2 to 3.0 J/cm2). One PL flash at 3 J/cm2 allowed at least 3 log-reduction of all tested microorganisms. The emetic B. cereus strain F4810/72 was the most resistant of the tested spore-forming bacteria. The PL fluence to 3 log-reduction (F3 PL) of its spores suspended in water was 2.9 J/cm2 and F3 UVC was 0.21 J/cm2, higher than F3 PL and F3 UVC of spores of B. pumilus SAFR-032 2.0 J/cm2 and 0.15 J/cm2, respectively), yet reported as a highly UV-resistant spore-forming bacterium. PL and UVC sensitivity of bacterial spores was correlated. Aspergillus spp. conidia suspended in water were poorly sensitive to PL. In contrast, PL inactivated Aspergillus spp. conidia spread on a dry surface more efficiently than UVC. The F2 PL of A. brasiliensis DSM1988 was 0.39 J/cm2 and F2 UVC was 0.83 J/cm2. The resistance of spore-forming bacteria to PL could be reasonably predicted from the knowledge of their UVC resistance. In contrast, the sensitivity of fungal conidia to PL must be specifically explored.

Biochemical Characterization of a Novel Thermostable 1,4-α-Glucoamylase from Aspergillus brasiliensis Strain Isolated in the Brazilian Atlantic Forest.

Glucoamylases are exo-enzymes that cleave the ends of the starch chain, releasing glucose units. In the current work, we described a novel 1,4-α-glucoamylase from an A. brasiliensis strain isolated from an environmental sample. The purified glucoamylase, GlaAb, has a molecular mass of 69 kDa and showed a starch binding domain. GlaAb showed a similar sequence to other fungal glucoamylases, and the molecular 3D model analysis of GlaAb suggests an overall structure as described in the literature, except by elongation in the loop connecting the 4th and 5th α-helices. The enzyme showed activity over a wide range of pH and temperature, with maximum activity at pH 4.5 and 60 °C. GlaAb was stable at 50 °C for 7 h, maintaining 67% residual activity, and it was not inhibited by glucose up to 0.1 M. The glucoamylase was 65% more active in the presence of Mn2+ and showed a Km of 2.21 mg mL-1, Vmax of 155 U mg-1, Kcat 179 s-1, and Kcat/Km 81.06 mg mL-1 s-1 using potato starch as substrate. The results obtained are promising and provide the basis for the development of applications of GlaAb in the industrial process.

Biomimetic Nanopillar Silicon Surfaces Rupture Fungal Spores.

The mechano-bactericidal action of nanostructured surfaces is well-documented; however, synthetic nanostructured surfaces have not yet been explored for their antifungal properties toward filamentous fungal species. In this study, we developed a biomimetic nanostructured surface inspired by dragonfly wings. A high-aspect-ratio nanopillar topography was created on silicon (nano-Si) surfaces using inductively coupled plasma reactive ion etching (ICP RIE). To mimic the superhydrophobic nature of insect wings, the nano-Si was further functionalised with trichloro(1H,1H,2H,2H-perfluorooctyl)silane (PFTS). The viability of Aspergillus brasiliensis spores, in contact with either hydrophobic or hydrophilic nano-Si surfaces, was determined using a combination of standard microbiological assays, confocal laser scanning microscopy (CLSM), and focused ion beam scanning electron microscopy (FIB-SEM). Results indicated the breakdown of the fungal spore membrane upon contact with the hydrophilic nano-Si surfaces. By contrast, hydrophobised nano-Si surfaces prevented the initial attachment of the fungal conidia. Hydrophilic nano-Si surfaces exhibited both antifungal and fungicidal properties toward attached A. brasisiensis spores via a 4-fold reduction of attached spores and approximately 9-fold reduction of viable conidia from initial solution after 24 h compared to their planar Si counterparts. Thus, we reveal, for the first time, the physical rupturing of attaching fungal spores by biomimetic hydrophilic nanostructured surfaces.

Biorefinery of Brewery Spent Grain by Solid-State Fermentation and Ionic Liquids.

Novel environmentally friendly pretreatments have been developed in recent years to improve biomass fractionation. Solid-state fermentation (SSF) and treatment with ionic liquids show low environmental impact and can be used in biorefinery of biomass. In this work, these processes were assessed with brewery spent grain (BSG). First, BSG was used as a substrate to produce cellulases and xylanases by SSF with the fungi Aspergillus brasiliensis CECT 2700 and Trichoderma reesei CECT 2414. Then, BSG was pretreated with the ionic liquid [N1112OH][Gly] and hydrolyzed with the crude enzymatic extracts. Results showed that SSF of BSG with A. brasiliensis achieved the highest enzyme production; meanwhile, the pretreatment with ionic liquids allowed glucan and xylan fractions to increase and reduce the lignin content. In addition, a mixture of the extracts from both fungi in a ratio of 2.5:0.5 Aspergillus/Trichoderma (v/v) efficiently hydrolyzed the BSG previously treated with the ionic liquid [N1112OH][Gly], reaching saccharification percentages of 80.68%, 54.29%, and 19.58% for glucan, xylan, and arabinan, respectively. In conclusion, the results demonstrated that the BSG biorefinery process developed in this work is an effective way to obtain fermentable sugar-containing solutions, which can be used to produce value-added products.

Antifungal Activity of Soft Tissue Extract from the Garden Snail Helix aspersa (Gastropoda, Mollusca).

Gastropods comprise approximately 80% of molluscans, of which land snails are used variably as food and traditional medicines due to their high protein content. Moreover, different components from land snails exhibit antimicrobial activities. In this study, we evaluated the antifungal activity of soft tissue extracts from Helix aspersa against Candida albicans, Aspergillus flavus, and Aspergillus brasiliensis by identifying extract components using liquid chromatography-tandem mass spectrometry (LC-MS-MS). Two concentrations of three extracts (methanol, acetone, and acetic acid) showed antifungal activity. Both acetone (1 g/3 mL) and acetic acid extracts (1 g/mL) significantly inhibited C.albicans growth (p = 0.0001, 5.2 ± 0.2 mm and p = 0.02, 69.7 ± 0.6 mm, respectively). A. flavus and A. brasiliensis growth were inhibited by all extracts at 1 g/mL, while inhibition was observed for acetic acid extracts against A. brasiliensis (p = 0.02, 50.3 ± 3.5 mm). The highest growth inhibition was observed for A. flavus using acetic acid and acetone extracts (inhibition zones = 38 ± 1.7 mm and 3.1 ± 0.7 mm, respectively). LC-MS-MS studies on methanol and acetone extracts identified 11-α-acetoxyprogesterone with a parent mass of 372.50800 m/z and 287.43500 m/z for luteolin. Methanol extracts contained hesperidin with a parent mass of 611.25400 m/z, whereas linoleic acid and genistein (parent mass = 280.4 and 271.48900 m/z, respectively) were the main metabolites.

Ent-hardwickiic acid from C. pubiflora and its microbial metabolites are more potent than fluconazole in vitro against Candida glabrata.

The incidence of Candida glabrata infections has rapidly grown and this species is among those responsible for causing invasive candidiasis with a high mortality rate. The diterpene ent-hardwickiic acid is a major constituent in Copaifera pubiflora oleoresin and the ethnopharmacological uses of this oleoresin by people from Brazilian Amazonian region point to a potential use of this major constituent as an antimicrobial. Therefore, the goal of this study was to evaluate the antifungal activity of ent-hardwickiic acid against Candida species and to produce derivatives of this diterpene by using microbial models for simulating the mammalian metabolism. The microbial transformations of ent-hardwickiic acid were carried out by Aspergillus brasiliensis and Cunninghamella elegans and hydroxylated metabolites were isolated and their chemical structures were determined. The antifungal activity of ent-hardwickiic acid and its metabolites was assessed by using the microdilution broth method in 96-well microplates and compared with that of fluconazole. All the diterpenes showed fungistatic effects (ranging from 19·7 to 75·2 µmol l-1 ) against C. glabrata at lower concentrations than fluconazole (163·2 µmol l-1 ) and were more potent fungicides (ranging from 39·5 to 150·4 µmol l-1 ) than fluconazole, which showed fungicidal effect at the concentration of 326·5 µmol l-1 .

Sterilization of food packaging by UV-C irradiation: Is Aspergillus brasiliensis ATCC 16404 the best target microorganism for industrial bio-validations?

In food industries UV-C irradiation is used to achieve decontamination of some packaging devices, such as plastic caps or laminated foils, and of those smooth surfaces that can be directly irradiated. Since its effectiveness can be checked by microbial validation tests, some ascospore-forming molds (Aspergillus hiratsukae, Talaromyces bacillisporus, Aspergillus montevidensis, and Chaetomium globosum) were compared with one of the target microorganisms actually used in industrial bio-validations (Aspergillus brasiliensis ATCC 16404) to find the species most resistant to UV-C. Tests were carried out with an UV-C lamp (irradiance = 127 µW/cm2; emission peak = 253.7 nm) by inoculating HDPE caps with one or more layers of spores. Inactivation kinetics of each strain were studied and both the corresponding 1D-values and the number of Logarithmic Count Reductions (LCR) achieved were calculated. Our results showed the important role played by the type of inoculum (one or more layers) and by the differences in cell structure (thickness, presence of protective solutes, pigmentation, etc.) of the strains tested. With a single-layer inoculum, Chaetomium globosum showed the highest resistance to UV-C irradiation (1D-value = 100 s). With a multi-layer inoculum, Aspergillus brasiliensis ATCC 16404 was the most resistant fungus (1D-value = 188 s), even if it reached a number of logarithmic reductions that was higher than those of some ascospore-forming mycetes (Aspergillus montevidensis, Talaromyces bacillisporus) tested.

Comparison of Different Temperature Conditions for Microbial Enumeration of Aspergillus brasiliensis in the Preservative Efficacy Test.

The preservative efficacy test is an important method for assessing the antimicrobial effect of cosmetic products. In this study, the optimum conditions for the efficient microbial enumeration of Aspergillus brasiliensis were investigated. Cosmetic products, inoculated with A. brasiliensis spore suspensions, were cultivated at 22.5°C, 32.5°C, or 40°C and the detection rate and the number of colonies were determined using the pour culture method. There was no difference in the viable counts of visible colonies among different temperature conditions. However, the viable counts after 3 days of culture were significantly greater for the cultures maintained at 32.5°C or 40°C compared with those maintained at 22.5°C. This effect was attenuated in products containing fatty acids, which could inhibit fungal growth. Overall, these results demonstrate that cultivating A. brasiliensis at 32.5°C reduces the time required for enumeration in the preservative efficacy test. Thus, the results of this study are expected to help improve and expedite microbiological quality control in the cosmetic industry.

Influence of type, concentration, exposure time, temperature, and presence of organic load on the antifungal efficacy of industrial sanitizers against Aspergillus brasiliensis (ATCC 16404).

Parameters such as type and concentration of the active compound, exposure time, application temperature, and organic load presence influence the antimicrobial action of sanitizers, although there is little data in the literature. Thus, this study aimed to evaluate the antifungal efficacy of different chemical sanitizers under different conditions according to the European Committee for Standardization (CEN). Aspergillus brasiliensis (ATCC 16404) was exposed to four compounds (benzalkonium chloride, iodine, peracetic acid, and sodium hypochlorite) at two different concentrations (minimum and maximum described on the product label), different exposure times (5, 10, and 15 min), temperatures (10, 20, 30, and 40 °C), and the presence or absence of an organic load. All parameters, including the type of sanitizer, influenced the antifungal efficacy of the tested compounds. Peracetic acid and benzalkonium chloride were the best antifungal sanitizers. The efficacy of peracetic acid increased as temperatures rose, although the opposite effect was observed for benzalkonium chloride. Sodium hypochlorite was ineffective under all tested conditions. In general, 5 min of sanitizer exposure is not enough and >10 min are necessary for effective fungal inactivation. The presence of organic load reduced sanitizer efficacy in most of the tested situations, and when comparing the efficacy of each compound in the presence and absence of an organic load, a difference of up to 1.5 log CFU was observed. The lowest concentration recommended on the sanitizer label is ineffective for 99.9% fungal inactivation, even at the highest exposure time (15 min) or under the best conditions of temperature and organic load absence. Knowledge of the influence exerted by these parameters contributes to successful hygiene since the person responsible for the sanitization process in the food facility can select and apply a certain compound in the most favorable conditions for maximum antifungal efficacy.

Potentiated inhibition of Trichoderma virens and other environmental fungi by new biocide combinations.

Fungi cause diverse, serious socio-economic problems, including biodeterioration of valuable products and materials that spawns a biocides industry worth ~$11 billion globally. To help combat environmental fungi that commonly colonise material products, this study tested the hypothesis that combination of an approved fungicide with diverse agents approved by the FDA (Food and Drug Administration) could reveal potent combinatorial activities with promise for fungicidal applications. The strategy to use approved compounds lowers potential development risks for any effective combinations. A high-throughput assay of 1280 FDA-approved compounds was conducted to find those that potentiate the effect of iodopropynyl-butyl-carbamate (IPBC) on the growth of Trichoderma virens; IPBC is one of the two most widely used Biocidal Products Regulations-approved fungicides. From this library, 34 compounds in combination with IPBC strongly inhibited fungal growth. Low-cost compounds that gave the most effective growth inhibition were tested against other environmental fungi that are standard biomarkers for resistance of synthetic materials to fungal colonisation. Trifluoperazine (TFZ) in combination with IPBC enhanced growth inhibition of three of the five test fungi. The antifungal hexetidine (HEX) potentiated IPBC action against two of the test organisms. Testable hypotheses on the mechanisms of these combinatorial actions are discussed. Neither IPBC + TFZ nor IPBC + HEX exhibited a combinatorial effect against mammalian cells. These combinations retained strong fungal growth inhibition properties after incorporation to a polymer matrix (alginate) with potential for fungicide delivery. The study reveals the potential of such approved compounds for novel combinatorial applications in the control of fungal environmental opportunists. KEY POINTS: • Search with an approved fungicide to find new fungicidal synergies in drug libraries. • New combinations inhibit growth of key environmental fungi on different matrices. • The approach enables a more rapid response to demand for new biocides.

Effect of peracetic acid on ascospore-forming molds and test microorganisms used for bio-validations of sanitizing processes in food plants.

Industrial sterilization of packaging and filling machineries by peracetic acid (PAA) is a widespread practice. In our study we assessed the resistance to PAA of three ascospore-forming molds (Chaetomium globosum ATCC 6205; Talaromyces bacillisporus SSICA 10915; Aspergillus hiratsukae SSICA 3913) compared to that of Aspergillus brasiliensis ATCC 16404 and Bacillus atrophaeus DSM 675, that are currently used as test microorganisms during industrial bio-validations of food packaging and machineries. Tests were carried out at 40 °C using 1,000 mg/l of PAA, with or without a supporting material (aluminium, tin-plate, PET). At all conditions tested, a greater resistance to PAA was registered for C. globosum, followed by T. bacillisporus, A. hiratsukae, A. brasiliensis and B. atrophaeus. D-values of C. globosum varied from 23 to 68 min, whereas T. bacillisporus showed D-values from 83 to 352 s and A. hiratsukae showed D-values from 32 to 65 s. Surprisingly, both test microorganisms (A. brasiliensis and B. atrophaeus) proved less resistant than ascospore-forming molds tested, their D-values being always lower than 30 s. Cells treated without a supporting material proved more resistant than those deposited on plastic or metallic strips, with the exception of tin-plate, where results approaching those obtained without a supporting materials were obtained. Based on the results obtained in this paper, test microorganisms currently used for bio-validations in industrial plants and also heat-resistant strains proved sensibly less resistant to PAA than C. globosum. Therefore, for practical purposes C. globosum should be furtherly studied to understand if its use during bio-validations of sanitizing processes could lead to more performing results.

Proximal subungual onychomycosis of digitus minimus due to Aspergillus brasiliensis.

Onychomycosis is a common nail infection caused by dermatophytes, non-dermatophytic molds (NDMs) and yeast. Aspergillus spp. are emerging etiological agents of non-dermatophyte mold onychomycosis (NDMO). Though this is usually of cosmetic concern, it may also cause pain and discomfort to the patient. The toenail is more commonly involved as compared to fingernail. The nails are discoloured and disfigured. Onychomycosis may expose the patient to cellulitis of lower extremities. The clinical presentation of dermatophytic and NDM onychomycosis is more or less similar, which creates problem in the diagnosis. Fingernail infection may cause social and psychological problem to the patient if fingernail is involved. Incidence of onychomycosis has been seen more in immunosuppressed individuals, where it is of more serious medical concern. In the present study we are reporting a case of proximal subungual onychomycosis (PSO) due to Aspergillus brasiliensis.

Fungistatic and Fungicidal Capacity of a Biosurfactant Extract Obtained from Corn Steep Water.

Biosurfactants are surface-active compounds that are produced by microorganisms, which in addition to their surfactant capacity, can possess interesting antimicrobial activities that are used in their incorporation into the agrifood industry. In this work, the preservative capacity of a novel biosurfactant extract obtained from a residual stream of the corn-milling industry was evaluated against two different fungi (Aspergillus brasiliensis and Candida albicans) under different biosurfactant concentrations (0.33-0.99 mg/mL), temperatures (4-40 °C), and incubation times (5-11 days). All the assays started with the same concentration of fungi (2 × 106 CFU/mL). The results showed that temperature played an important role in the bactericidal and fungistatic effects of this biosurfactant extract. It was observed that at a low biosurfactant concentration (0.33 mg/mL) and low or high temperatures in the range tested, this biosurfactant extract possessed an important fungicidal effect (complete inhibition) on A. brasiliensis, while at intermediate temperatures, it achieved a fungistatic effect (50% of inhibition). Regarding C. albicans, it was observed that this strain was more resistant than A. brasiliens, although it was possible to achieve growth inhibitions of 76.3% at temperatures of 40 °C after 8 days of incubation with a biosurfactant concentration of 0.99 mg/mL. This work supports the possible application of biosurfactants extracted from corn steep water as preservatives and antimicrobial agents against fungal contaminations on agrifood products.

New antifungal ent-labdane diterpenes against Candida glabrata produced by microbial transformation of ent-polyalthic acid.

Candida glabrata, the most common non-albicans Candida species and one of the primary causes of candidemia, exhibits decreased susceptibility to azoles and more recently to echinocandins. Polyalthic acid 1, a furan diterpene, has been shown promising biological potential and in this study ent-polyalthic acid derivatives with antifungal activity against Candida glabrata were produced by microbial transformation. Incubation of 1 with Aspergillus brasiliensis afforded two known (compounds 5 and 10) and eight new derivatives (compounds 2-4, 6-9 and 11). The most common reaction was hydroxylation, but isomerization of the double bond and acetylation were also detected. None of the tested compounds showed cytotoxicity against HeLa, MCF-7 and MCF-10A cell lines showing IC50 values ranging from 62.6 µM to > 500 µM. Compounds 1, 5, 6, 8 and 11 showed fungistatic effects (ranging from 34.1 µM to 39.5 µM) on C. glabrata at lower concentrations than fluconazole (163.2 µM). Compounds 1, 6 and 8 were more potent fungicides (ranging from 79.0 to 143.6 µM) than fluconazole, which showed fungicidal effect at concentrations higher than 163.2 µM. These results suggest that ent-polyalthic acid and some of its derivatives could be used as lead compounds to develop new antifungal agents.

Sanitization of packaging and machineries in the food industry: Effect of hydrogen peroxide on ascospores and conidia of filamentous fungi.

In the food industry, sterilization of packaging and filling machines by hydrogen peroxide (HP) is a widespread practice. Its effectiveness is usually tested by means of inactivation tests on selected test microorganisms that were any case chosen without taking into account that food products could be also spoiled by microorganisms presumably resistant to HP. For this reason, the aim of this work was to assess the resistance of different ascospore-forming moulds (Talaromyces bacillisporus, Aspergillus hiratsukae, Chaetomium globosum) to HP, in order to find the most resistant to this kind of chemical stress, and to compare their resistance with that registered for other moulds, including test microorganism Aspergillus brasiliensis ATCC 16404. Tests were carried out from 50 to 60 °C on spores or conidia, depending on the strain, either by immersing inoculated strips (aluminium, tin-plate, HDPE, PET) in HP, or by directly inoculating cells in the sanitizing medium. In both tests, T. bacillisporus proved the most resistant strain, followed by A. hiratsukae, C. globosum and A. brasiliensis at all temperatures tested. In test without a supporting material, D values of T. bacillisporus varied from 6 to 23 s. In test with metallic or plastic strips, D values of T. bacillisporus were higher on plastic materials, compared to those obtained on metallic ones up to 53 °C, whereas at higher temperatures D values proved similar. For A. hiratsukae, D values were similar if different materials were compared, except for D50 on aluminium and HDPE, which proved slightly higher (3.1-3.4 s) than those obtained on tin-plate and PET (2.7-2.8 s). Analogously, ascospores of C. globosum behaved in a similar way if different materials were compared, except for D50 values that varied in a wide range (from 2.9 s on tin-plate to 4.0 s on HDPE). A. brasiliensis was rapidly inactivated by the synergistic effect of heat and hydrogen peroxide, so for this strain it was not possible to calculate any D value. Based on the results obtained in this paper, tested ascospore-forming moulds proved to be sensibly more resistant to HP than other heat-sensitive strains tested, their D values always being significantly higher, regardless of the strain considered and the supporting material assessed. Ascospore-forming moulds could be furtherly investigated, as for practical purposes they seemed most suitable as target microorganisms than heat-sensitive microorganisms such as Aspergillus brasiliensis ATCC 16404, their use during bio-validations of sanitizing processes on machineries used for refrigerated products (pH > 4.5) or non-refrigerated acid products (pH ≤ 4.5) leading to more performing results.

Preservative and Irritant Capacity of Biosurfactants From Different Sources: A Comparative Study.

One of the most important challenges for pharmaceutical and cosmetic industries is solubilization and preservation of their active ingredients. Therefore, most of these formulations contain irritant chemical additives to improve their shelf-life and the solubility of hydrophobic ingredients. An interesting alternative to chemical surfactants and preservatives is the use of biosurfactants; thus, their surfactant properties and composition make them more biocompatible than their chemical counterparts. Moreover, some biosurfactants have shown antimicrobial activity in addition to their detergent capacity. In this work, the antimicrobial and irritant effect of 2 biosurfactant extracts was studied: one produced in a controlled fermentation process with Lactobacillus pentosus and the other produced from corn stream by spontaneous fermentation. The results showed a strong antimicrobial activity of the biosurfactant extract obtained from corn stream on pathogenic bacteria, in comparison with the L. pentosus biosurfactant extract. Moreover, both biosurfactants did not produce any irritant effect on the chorioallantoic membrane of hen's egg assay contrary to sodium dodecyl sulfate. This is the first study dealing with the application of biosurfactant extracts on sensitive biological membranes, and this is the first time that the preservative capacity of a biosurfactant extract obtained in spontaneous fermentation is being evaluated, achieving promising results.

Characterization and antimicrobial activity of silver nanoparticles mycosynthesized by Aspergillus brasiliensis.

AIMS: Since mycosynthesis of metal nanoparticles (NPs) is advertised as a promising and ecofriendly approach. Thus, this study aims to investigate the capability of Aspergillus brasiliensis ATCC 16404 for mycosynthesis of silver NPs (AgNPs). METHODS AND RESULTS: One-factor-at-a-time-technique was used to study the effect of different physicochemical parameters: the reaction time, pH, temperature, different stirring rates, illumination, and finally, the different concentrations of silver nitrate and fungal biomass on the mycosynthesis of AgNPs. The visual observation showed the characteristic brown colour formation due to the bioreduction of Ag+ ions to Ag0 by the mycelial cell-free filtrate (MCFF). The UV/visible spectrophotometric technique displayed a characteristic sharp peak at ʎ440 confirming the mycosynthesis of AgNPs. The zeta potential value -16·7 mV assured the long-term stability of AgNPs and the dynamic light scattering analysis revealed good dispersion and average particle size 77 nm. The energy dispersive X-ray spectroscopy displayed a maximum elemental distribution of silver elements. The X-ray diffraction spectroscopy demonstrated the crystallinity of the mycosynthesized AgNPs. The field emission scanning electron microscope and high-resolution transmission electron microscope revealed monodispersed spherical shaped AgNPs with average particle size of 6-21 nm. The FTIR analysis showed the major peaks of proteins providing the possible role of MCFF in the synthesis and stabilization of the AgNPs. The mycosynthesized AgNPs expressed good biocidal activity against different pathogenic micro-organisms causing some water-related diseases and health problems to local residents. CONCLUSIONS: This study proved that A. brasiliensis ATCC 16404 MCFF has good potential for mycosynthesis of AgNPs, which exhibited good antimicrobial effect on different pathogenic micro-organisms; thus, it can be applied for water disinfection. SIGNIFICANCE AND IMPACT OF THE STUDY: This research provides a helpful insight into the development of a new mycosynthesized antimicrobial agent.

Polycyclic aromatic hydrocarbon-emulsifier protein produced by Aspergillus brasiliensis (niger) in an airlift bioreactor following an electrochemical pretreatment.

An emulsifier protein (EP) was produced and easily separated from oil-contaminated water as an economical substrate when Aspergillus brasiliensis, pretreated in a solid state culture with a controlled electric field, was used in an airlift bioreactor. The hydrocarbon-EP comprised 19.5% of the total protein, its purification enhanced the specific emulsifying activity (EA) seven times. The influence of operational conditions (pH and salt concentration) on the EA were assessed to characterise the emulsion stability. The EA was increased by 19% in alkaline environments (pH 7-11), but it was not affected by the presence of salt (0-35 g L-1). On the other hand, preheating the EP samples (60 °C) enhanced the EA by 2.5 times. Based on analysis of its EA, this EP can be applied as a bioremediation enhancer in contaminated soils.