Biocoagulation of Dried Algae Chlorella sp. and Pellets of Aspergillus Niger in Decontamination Process of Wastewater, as a Presumed Source of Biofuel.

The removal of microalgae represents a problematic part of the water decontamination process, in which most techniques are expensive and non-ecological. In the paper, we focus on the synergistic relationship between microscopic filamentous fungi and algal culture. In the process of decontamination of a model sample containing ammonium ions, efficient biocoagulation, resp. co-pelletization of dried algae Chlorella sp. and Aspergillus niger sensu stricto are shown. The microscopic filamentous fungus species A. niger was added to a culture of an algal suspension of Chlorella sp., where the adhesion of the algal cells to the fungi subsequently occurred due to the electrostatic effect of the interaction, while the flocculation activity was approximately 70 to 80%. The algal cells adhered to the surface of the A. niger pellets, making them easily removable from the solution. The ability of filamentous fungi to capture organisms represents a great potential for the biological isolation of microalgae (biocoagulation) from production solutions because microalgae are considered to be a promising renewable source of oil and fermentables for bioenergy. This form of algae removal, or its harvesting, also represents a great low-cost method for collecting algae not only as a way of removing unnecessary material but also for the purpose of producing biofuels. Algae are a robust bioabsorbent for absorbing lipids from the environment, which after treatment can be used as a component of biodiesel. Chemical analyses also presented potential ecological innovation in the area of biofuel production. Energy-efficient and eco-friendly harvesting techniques are crucial to improving the economic viability of algal biofuel production.

Experimental Treatment of Hazardous Ash Waste by Microbial Consortium Aspergillus niger and Chlorella sp.: Decrease of the Ni Content and Identification of Adsorption Sites by Fourier-Transform Infrared Spectroscopy.

Despite the negative impact on the environment, incineration is one of the most commonly used methods for dealing with waste. Besides emissions, the production of ash, which usually shows several negative properties, such as a higher content of hazardous elements or strongly alkaline pH, is problematic from an environmental viewpoint as well. The subject of our paper was the assessment of biosorption of Ni from ash material by a microbial consortium of Chlorella sp. and Aspergillus niger. The solid substrate represented a fraction of particles of size <0.63 mm with a Ni content of 417 mg kg-1. We used a biomass consisting of two different organisms as the sorbent: a non-living algae culture of Chlorella sp. (an autotrophic organism) and the microscopic filamentous fungus A. niger (a heterotrophic organism) in the form of pellets. The experiments were conducted under static conditions as well as with the use of shaker (170 rpm) with different modifications: solid substrate, Chlorella sp. and pellets of A. niger; solid substrate and pellets of A. niger. The humidity-temperature conditions were also changed. Sorption took place under dry and also wet conditions (with distilled water in a volume of 30-50 ml), partially under laboratory conditions at a temperature of 25°C as well as in the exterior. The determination of the Ni content was done using inductively coupled plasma optical emission spectrometry (ICP-OES). The removal of Ni ranged from 13.61% efficiency (Chlorella sp., A. niger with the addition of 30 ml of distilled water, outdoors under static conditions after 48 h of the experiment) to 46.28% (Chlorella sp., A. niger with the addition of 30 ml of distilled water, on a shaker under laboratory conditions after 48 h of the experiment). For the purpose of analyzing the representation of functional groups in the microbial biomass and studying their interaction with the ash material, we used Fourier-transform infrared (FTIR) spectroscopy. We observed that the amount of Ni adsorbed positively correlates with absorbance in the spectral bands where we detect the vibrations of several organic functional groups. These groups include hydroxyl, aliphatic, carbonyl, carboxyl and amide structural units. The observed correlations indicate that, aside from polar and negatively charged groups, aliphatic or aromatic structures may also be involved in sorption processes due to electrostatic attraction. The correlation between absorbance and the Ni content reached a maximum in amide II band (r = 0.9; P < 0.001), where vibrations of the C=O, C-N, and N-H groups are detected. The presented results suggest that the simultaneous use of both microorganisms in biosorption represents an effective method for reducing Ni content in a solid substrate, which may be useful as a partial process for waste disposal.

Aspergillus niger Environmental Isolates and Their Specific Diversity Through Metabolite Profiling.

We present a biological profile of 16 Aspergillus niger environmental isolates from different types of soils and solid substrates across a pH range, from an ultra-acidic (<3.5) to a very strongly alkaline (>9.0) environment. The soils and solid substrates also differ in varying degrees of anthropic pollution, which in most cases is caused by several centuries of mining activity at old mining sites, sludge beds, ore deposits, stream sediments, and coal dust. The values of toxic elements (As, Sb, Zn, Cu, Pb) very often exceed the limit values. The isolates possess different macro- and micromorphological features. All the identifications of Aspergillus niger isolates were confirmed by molecular PCR analysis and their similarity was expressed by RAMP analysis. The biochemical profile of isolates based on FF-MicroPlate tests from the Biolog system showed identical biochemical reactions in 50 tests, while in 46 tests the utilisation reactions differed. The highest similarity of strains isolated from substrates with the same pH, as well as the most suitable biochemical tests for analysis of the phenotypic similarity of isolated strains, were confirmed when evaluating the biochemical profile using multicriterial analysis in the Canoco program. The isolates were screened for mycotoxin production by thin-layer chromatography (TLC), as well. Two of them were able to synthesise ochratoxin A, while none produced fumonisins under experimental conditions. Presence of toxic compounds in contaminated sites may affect environmental microscopic fungi and cause the genome alteration, which may result in changes of their physiology, including the production of different (secondary) metabolites, such as mycotoxins.

Occurrence and diversity of cultivable autochthonous microscopic fungi in substrates of old environmental loads from mining activities in Slovakia.

Distribution and biodiversity of soil microscopic fungi in 5 areas of old environmental loads in Slovakia were studied in relation to very low amount of organic matter (%TOC from 0.2 to 3.54) and to the pH gradient from ultra-acidic (< 3.5) to very strongly alkaline (> 9.0). All soil samples were affected by several hundred years of mining activities and contained heavy metals and other toxic elements: arsenic, cadmium, copper, zinc, antimony, lead. Concentrations of toxicants highly exceeded their limited values. Fifty-three genera and 112 species of microscopic fungi were identified. Among them, Zygomycota occurred very rarely (8 genera and 12 species), except of samples with the highest content of TOC (2.01-3.54% - samples 2 and 6), regardless their pH. Though, on the other hand, from some similar samples (3, 5 and 9), incl. those with relatively high TOC (0.14-2.62%), the lower fungi were not recovered. Forty one genera and 95 species of Ascomycota represented the most abundant fungal phylum in all investigated samples. Among them, Penicillium chrysogenum var. chrysogenum, Aspergillus niger and Neosartorya fischeri were isolated the most often. Phytopathogenic moulds of Bionectria ochroleuca, Lewia infectoria, Phoma macrostoma and Phlebia acerina were also occurred frequently. The highest biodiversity of microfungal community was recorded in the extreme acidic environment, followed by the neutral, ultra-acidic and the very strong acidic ones. There was no similarity in microfungal spectrum found in the samples studied. Except of the ultra acidic and extreme acidic samples (1-2) as well as the ultra acidic and strong acidic ones (1-4) with the most rich mycobiota, that may indicate a certain similarity degree.

Removal of aluminium from aqueous solution by four wild-type strains of Aspergillus niger.

This paper provides a unique comparison of the performance of four wild-type Aspergillus niger strains in remediation of aluminium(III)-contaminated aqueous solutions. The direct fungal aluminium removal via biosorption and bioaccumulation was compared among all fungal strains, including bioaccumulation efficiency during dynamic and static cultivation. Our results indicate that aluminium bioaccumulation by living biomass outperformed biosorption, although biosorption by non-living biomass is a less time-demanding process. Among others, only one strain significantly differed regarding comparison of dynamic and static bioaccumulation. In this case, a significantly higher removal performance was achieved under dynamic cultivation conditions at initial aluminium(III) concentrations over 2.5 mg L-1. Although the fungal sensitivity towards aluminium(III) differed among selected fungal strains, there was no apparent correlation between the strains' removal performance and their adaptive mechanisms.

Disclosing a crypt: microbial diversity and degradation activity of the microflora isolated from funeral clothes of Cardinal Peter Pázmány.

A crypt can be considered as a particular environment where different microbial communities contribute to decomposition of organic materials present inside during a long interval of time. The textile remains of the funeral clothes (biretta and tunic) of Cardinal Pázmány, an important historic figure dead in Bratislava the 19th March 1637, conserved in this kind of environment were subjected to microbial investigation. The sampling comprised three different approaches and the use of various kinds of cultivation media. Two different PCR-based clustering methods, f-ITS and f-CBH, were employed in order to select the bacterial and fungal microfloras which were identified in a second step by the 16S rRNA and ITS sequencing respectively. The isolated microflora was tested for its proteolytic, keratinolytic and cellulolytic activities and for its ability to grow on Fibroin agar medium. The combination of cultural, molecular and biodegradative assays was able to isolate and characterize a bacterial community composed mainly by members of the phyla Firmicutes and Actinobacteria. The fungal community appeared more diversified, together with several Penicillium and Aspergillus strains, members belonging to the species Beauveria bassiana, Eurotium cristatum, Xenochalara juniperi, Phialosimplex caninus and Myriodontium keratinophilum were isolated. Bacteria, especially the Bacillus members, showed their strong ability to degrade keratin and gelatin and a large portion of them was able to growth on Fibroin agar. The fungal isolates displayed a widespread cellulolytic activity and fibroin utilization, although they possessed a weaker and slower proteolytic and keratinolytic properties respect to bacterial counterpart. The present study can be considered perhaps as the first or among the few microbial investigations which treated the textile biodegradation from such unusual environment.

Influence of the Environment on the Morphological and Biochemical Characteristics of Different Aspergillus niger Wild Type Strains.

The present work studied the differences in accumulation, transformation and volatilization of different heavy metals ions on molecular and macromorphological features of Aspergillus niger wild type strains. Four different strains of A. niger (An) were used. Three strains (An-P, An-N, An-S) were isolated from acid and ultra acid mining regions with higher concentration of As and Sb. The fourth strain (An-G) was used as the comparative one. Environmental burden strongly affected biochemical, macro and micromorphological characteristics of studied strains. The RAMP profiles showed 90 % similarity among the studied strains. The strain An-S showed its own characteristic RAMP profile, different to the others ones. Analyzed strains can be clustered into two groups on the basis of the changes in gene expression and morphological parameters. Differences were found in both acid ß-1,3-glucanases and peroxidases. Main quantitative and qualitative differences by A-PAGE and SDS-PAGE were registered for proteins with Mr ~ 50; 34; 28-27 and 11 kDa. Presence of living mutants of A. niger strains in old environmental burden indicate on the adaptation and mutation processes of soil microorganisms from the point of long-term effect.

Analysis and comparison of the microflora isolated from fresco surface and from surrounding air environment through molecular and biodegradative assays.

The aim of this study was to find a correlation among the environmental isolated microflora and the fresco colonizators through the investigation of their biodegradative abilities and DNA characteristics. A molecular technique named RAMP (Random Amplified Microsatellite Polymorphisms) was utilized in order to analyze the DNA diversity of bacterial and fungal species isolated from fresco as well as from air samples. The RAMP-PCR results were combined with the screening of some biodegradative properties obtained through the use of specific agar plate assays detecting the proteolytic, solubilization and biomineralization abilities of the isolated microflora. This comparative analysis showed that only in few cases a direct link among the fresco and airborne isolates of specific microbial group existed. The investigation clearly evidenced that colonization of surface of Ladislav's fresco occurred in different time and by different strains than those observed at the moment of sampling campaign. Furthermore, the microflora investigation permitted the identification of taxonomically interesting bacteria with particular biodegradative properties, which had been less studied until now.

Sorption, desorption, and degradation of (4-chloro-2-methylphenoxy)acetic acid in representative soils of the Danubian Lowland, Slovakia.

Herbicide leaching through soil into groundwater greatly depends upon sorption-desorption and degradation phenomena. Batch adsorption, desorption and degradation experiments were performed with acidic herbicide MCPA and three soil types collected from their respective soil horizons. MCPA was found to be weakly sorbed by the soils with Freundlich coefficient values ranging from 0.37 to 1.03 mg(1-1/)(n) kg(-1) L(1/)(n). It was shown that MCPA sorption positively correlated with soil organic carbon content, humic and fulvic acid carbon contents, and negatively with soil pH. The importance of soil organic matter in MCPA sorption by soils was also confirmed by performing sorption experiments after soil organic matter removal. MCPA sorption in these treated soils decreased by 37-100% compared to the original soils. A relatively large part of the sorbed MCPA was released from soils into aqueous solution after four successive desorption steps, although some hysteresis occurred during desorption of MCPA from all soils. Both sorption and desorption were depth-dependent, the A soil horizons exhibited higher retention capacity of the herbicide than B or C soil horizons. Generally, MCPA sorption decreased in the presence of phosphate and low molecular weight organic acids. Degradation of MCPA was faster in the A soil horizons than the corresponding B or C soil horizons with half-life values ranging from 4.9 to 9.6 d in topsoils and from 11.6 to 23.4 d in subsoils.

Investigation of microbial community isolated from indoor artworks and air environment: identification, biodegradative abilities, and DNA typing.

This study deals with establishing the characteristics of a microbial community isolated from indoor artworks and the surrounding air environment. It is one of the few studies on microbial degradation of indoor artworks. It shows the potential biodegradative risk that can occur if artworks are not exhibited and conserved in an appropriate environment. The microbial community isolated from the indoor artworks and air environment was examined by cultural and molecular methods. Different plate assays were used to screen the biodegradative activity of the isolated microflora: Remazol Brilliant Blue R, phenol red, and Azure B for the ligninolytic properties; Ostazin brilliant red H-3B for cellulose degradation; CaCO3 glucose agar for solubilization activity; and B4 agar for biomineralization. To type the bacterial and fungal isolates, 2 PCR methods, repetitive extragenic palindromes (REP) and random amplified microsatellite polymorphisms (RAMP) were used. The art objects were principally colonized by fungi. The most commonly isolated strains were represented by hyphomycetes of the genera Penicillium, Aspergillus, Cladosporium, and Chaetomium. Members of these genera showed intensive biodegradation activity, both on wood and on stone. Bacteria were predominant in the air, exhibiting complex communities, both in the air and on the artworks. The most frequently isolated genera were Bacillus and Staphylococcus with extensive biodegradation abilities. REP-PCR revealed high variability within strains belonging to the same genus. RAMP is a new PCR-based method, used in this research for the first time to cluster the microfilamentous fungi and to characterize and select especially Penicillium and Aspergillus strains, which were isolated in a large number.

Production of catalases by Aspergillus niger isolates as a response to pollutant stress by heavy metals.

Isolates of Aspergillus niger, selected from the coal dust of a mine containing arsenic (As; 400 mg/kg) and from the river sediment of mine surroundings (As, 1651 mg/kg, Sb, 362 mg/kg), growing in minimal nitrate medium in the phase of hyphal development and spore formation, exhibited much higher levels of total catalase activity than the same species from the culture collection or a culture adapted to soil contaminated with As (5 mg/L). Electrophoretic resolution of catalases in cell-free extracts revealed three isozymes of catalases and production of individual isozymes was not significantly affected by stress environments. Exogenously added stressors (As(5+), Cd(2+), Cu(2+)) at final concentrations of 25 and 50 mg/L and H(2)O(2) (20 or 40 mM) mostly stimulated production of catalases only in isolates from mines surroundings, and H(2)O(2) and Hg(2+) caused the disappearance of the smallest catalase I. Isolates exhibited a higher tolerance of the toxic effects of heavy metals and H(2)O(2), as monitored by growth, than did the strain from the culture collection.