Respirometry for assessing the biodegradation of petroleum hydrocarbons.

The respiration method using the Micro-Oxymax respirometer was applied to evaluate the bioremediation potential of hydrocarbon-contaminated soils in two biopiles at the oil refinery in Czechowice-Dziedzice, Poland. In biopiles 1 and 2, two different technologies, i.e., enhanced (engineered) bioremediation and monitored natural attenuation (MNA) were used, respectively. In biopiles 1 and 2, the bioremediation process lasted 6 years and 8 months, respectively. The biodegradation of petroleum hydrocarbons was evaluated on the basis of CO2 production and O2 uptake. The CO2 production and O2 consumption rates during hydrocarbon biodegradation were calculated from the slopes of cumulative curve linear regressions. The results confirmed the hydrocarbon biodegradation process in both biopiles. However, in biopile 2 the process was more effective compared to biopile 1. In biopile 2, the O2 consumption and CO2 production means were 3.37 and 2.4 milliliters per kilogram of soil (dry weight) per minute, respectively. Whereas, in biopile 1, the O2 consumption and CO2 production means were 1.52 and 1.07 milliliters per kilogram of soil (dry weight) per minute, respectively. The mean biodegradation rate for biopile 2 was two times higher--67 mg hydrocarbons kg d.w.(-1)day(-1) compared with biopile 1, where the mean was 30 mg hydrocarbons kg d.w.(-1)day(-l). The results were correlated with petroleum hydrocarbon concentrations and microbial activity measured by dehydrogenase assay.

Use of molecular techniques in bioremediation.

In a practical sense, biotechnology is concerned with the production of commercial products generated by biological processes. More formally, biotechnology may be defined as "the application of scientific and engineering principles to the processing of material by biological agents to provide goods and services" (Cantor, 2000). From a historical perspective, biotechnology dates back to the time when yeast was first used for beer or wine fermentation, and bacteria were used to make yogurt. In 1972, the birth of recombinant DNA technology moved biotechnology to new heights and led to the establishment of a new industry. Progress in biotechnology has been truly remarkable. Within four years of the discovery of recombinant DNA technology, genetically modified organisms (GMOs) were making human insulin, interferon, and human growth hormone. Now, recombinant DNA technology and its products--GMOs are widely used in environmental biotechnology (Glick and Pasternak, 1988; Cowan, 2000). Bioremediation is one of the most rapidly growing areas of environmental biotechnology. Use of bioremediation for environmental clean up is popular due to low costs and its public acceptability. Indeed, bioremediation stands to benefit greatly and advance even more rapidly with the adoption of molecular techniques developed originally for other areas of biotechnology. The 1990s was the decade of molecular microbial ecology (time of using molecular techniques in environmental biotechnology). Adoption of these molecular techniques made scientists realize that microbial populations in the natural environments are much more diverse than previously thought using traditional culture methods. Using molecular ecological methods, such as direct DNA isolation from environmental samples, denaturing gradient gel electrophoresis (DGGE), PCR methods, nucleic acid hybridization etc., we can now study microbial consortia relevant to pollutant degradation in the environment. These techniques promise to provide a better understanding and better control of environmental biotechnology processes, thus enabling more cost effective and efficient bioremediation of our toxic waste and contaminated environments.

General assessment of the influence of a municipal landfill site and environmental factors on the occurrence of keratinolytic fungi in soil.

The study was to generally determine the influence of a municipal landfill site and environmental factors on the distribution of keratinolytic fungi in soil. The landfill site in Sosnowiec was selected for examination. Keratinolytic fungi occurred abundantly in soils of the landfill site examined and its surrounding area. Of 495 soil samples (Petri dishes) examined, 379 (76.56%) were found to be positive for keratinolytic fungi. Altogether, 1131 strains from 26 species were isolated from the samples. Among the fungi, some species with pathogenic properties (Microsporum racemosum, M. cookei, M. gypseum, Aphanoascus fulvescens and Scopulariopsis brevicaulis) were recorded. The influence of environmental factors on the qualitative and quantitative composition of keratinolytic fungi in the soils was complex. Among these factors, exchangeable acidity (pH in 1 M KCl, in particular), faecal bacterial contamination and the level of water deficit in soil were the most important. The conclusion has been drawn that municipal landfill sites are the sources of potentially pathogenic fungi with keratinolytic properties.

Studies on keratinophilic fungi. XI. Chrysosporium undulatum sp. nov.

Chrysosporium undulatum sp. nov., showing intermediate characteristics between Chrysosporium and Malbranchea, is described and illustrated. This taxon is characterised by long and curved to loosely coiled hyphal appendages, deflected to wavy fertile hyphae forming dense tufts and small, subglobose, frequently intercalary, verrucose conidia. Restriction enzyme analysis and comparison of sequences of the ITS1-5.8S-ITS2 ribosomal DNA region of the most similar species strongly support this proposal.

Effect of cadmium on growth of potentially pathogenic soil fungi.

The effect of Cd on the mycelial growth of some potentially pathogenic soil fungi was investigated. Sixty-four strains from twenty-five fungal species were tested for their susceptibility to Cd. Final colony diameter, radial growth rate and final dry mass of mycelium (for Cd: 1-200 ppm) were measured. EcD50 values were calculated from these parameters. The intra- and interspecific variability in the results obtained is presented. Among the keratinolytic fungal group, the general Arthrographis, trichophyton and Chrysosporium (including Chrysosporium anamorph of Aphanoascus reticulisporus) were the most resistant to Cd. T. mentagrophytes was the most sensitive of all Trichophyton species tested. Among the nonkeratinolytic fungal group, the genera Pseudallescheria, Absida and Rhizopus were highly resistant to Cd. Strains of Aspergillus were more resistant to Cd than strains of Penicillium. Zygomycetes were more resistant to Cd than Ascomycetes with Fungi Imperfecti. Nonkeratinolytic fungi showed higher resistance to Cd than keratinolytic fungi. The two last differences resulted from the extremely high EcD50 values for R. oryzae and A. corymbifera.

A study of keratinolytic fungi in mountain sediments. I. Hair-baiting data.

Sediments form mountain streams in Brenna (Beskid Slaski) were examined for keratinolytic fungi. A rare psychrophilic dermatophyte, Keratinophyton ceretanicus, occurred abundantly in the sediments. The qualitative and qualitative compositions of keratinolytic fungi depended on the water contamination with sewage and on the contents of plant organic material, small mineral particles and salts in the sediments.

A preliminary study of the occurrence of actidione-resistant fungi in sediments of Catalonian river mouths (Spain). I. Keratinolytic fungi and related Onygenales.

Sediments from eight river mouths along the Catalonian coast (Spain) were surveyed for keratinolytic fungi and related Onygenales. The actidione plating technique was employed. Of 532 actidione-treated sediment samples, 268 (50.3%) were positive for the fungi. Altogether, 384 fungal strains from 35 species were isolated from the samples. Narasimhella marginospora, Aphanoascus fulvescens, Neoarachnotheca keratinophila with its anamorph Myriodontium keratinophilum, Narasimhella hyalinospora, Beauveria alba, Sporothrix schenckii, Chrysosporium lobatum and Gymnoascus littoralis were the predominant species in sediments. Abundance of N. marginospora was clearly correlated with the degree of water pollution with sewage.

Studies on keratinophilic fungi. IX: Neoarachnotheca gen. nov. and a new species of Nannizziopsis.

Neoarachnotheca is proposed as a new genus of Onygenales. The outstanding generic characteristics are white, spherical ascomata with a wall formed by a network of hyphae and spherical, subhyaline ascospores with an irregular sheath. Nt. keratinophila, the type species, characterized by wavy peridial hyphae has been isolated from marine and river sediments and Myriodontium keratinophilum is its anamorph. Nannizziopsis tropicalis is proposed as a new species based on a strain isolated from soil in Burundi. RFLPs analysis of ITS and 5.8S rDNA support these proposals. The differences with related genera are discussed.

Molecular characterization, relatedness and antifungal susceptibility of the basidiomycetous Hormographiella species and Coprinus cinereus from clinical and environmental sources.

Hormographiella-like strains, isolated from different natural substrates and producing sclerotia and occasionally basidiomata of Coprinus cinereus, were compared morphologically and using molecular techniques with clinical strains of Hormographiella aspergillata and H. verticillata. Analysis of restriction fragment length polymorphisms of ribosomal and mitochondrial-like DNA confirmed interspecific differences between H. aspergillata and H. verticillata, supporting the morphological data, and helped demonstrate that H. aspergillata is the anamorph of C. cinereus. The latter was confirmed also by crossing tests. The analysis of the mtDNA restriction profiles revealed intraspecific variability in C. cinereus, which allowed differentiation of clinical and environmental strains. Due to the implication of C. cinereus and Hormographiella in human opportunistic infections, the antifungal susceptibility test is included. Results show that all strains were susceptible to miconazole, itraconazole and ketoconazole but not to flucytosine and fluconazol. Susceptibility against amphotericin B was variable; while H. verticillata was susceptible, four out of seven C. cinereus strains tested were resistant.

Interactions between selected geophilic fungi and pathogenic dermatophytes.

Interactions between selected geophilic fungi (keratinolytic and keratinophylic species) and pathogenic dermatophytes were examined in dual cultures on Sabouraud glucose agar (SGA). It was demonstrated that Trichophyton mentagrophytes var. mentagrophytes together with T. mentagrophytes var., quinceanum are the species most sensitive to the influences of the geophilic fungi. Microsporum persicolor was found to be more resistant to these influences. Only a few geophilic species inhibited the growth of M. canis on SGA. Keratinophilic species, unable to initiate hair decomposition but utilize the products of this process, exerted the larger inhibitory effect on the pathogenic dermatophytes than keratinolytic fungi.

A preliminary study on the occurrence of keratinolytic fungi in the street sweepings from Chorzów.

The street sweepings from the city of Chorzów were surveyed for keratinolytic fungi. Out of 106 Petri dishes examined, 98 (92.4%) were positive for these micro-organisms. Altogether, 185 fungal appearances belonging to 15 species were observed. Chrysosporium keratinophilum, Malbranchea flava, Ch. europae, Sporothrix schenckii, ch. anamorphs of Aphanoascus reticulisporus/fulvescens, Ch. an. Arthroderma curreyi, and M. an. Uncinocarpus reessi predominated in the sweepings. The occurrence of Ch. keratinophilum, Ch. europae, Ch. an. A. curreyi, and S. schenckii could depend on the content of heavy metals and the individual fractions in the sweepings. The epidemiological aspect of the presence of pathogenic fungi in the street dust was briefly discussed.

Sorption of cadmium by filamentous soil fungi.

The article presents the evaluation of the short-term sorption of Cd by selected soil fungi. The Freundlich adsorption isotherm method for metal sorption modelling was applied. Fungal strains belonging to two classes, Zygomycetes and Ascomycetes, were used. Altogether, six species from Ascomycetes (A. niger, A. nidulans, Ps. boydii, T. koningii, P. janthinellum, P. verrucosum) and three from Zygomycetes (R. rhizopodiformis, R. oryzae, R. pusillus) were examined. These fungi were selected for experiments due to the differences in their cell wall compositions. The walls of Ascomycetes and Zygomycetes are in the chitin-glucan and in the chitosan-chitin groups, respectively. It was found that the biomass of A. nidulans and R. pusillus displayed the relatively high Cd sorption capacity. The sorption of Cd was similar among the fungi investigated. The metal accumulation by these fungi increased with increasing concentrations of Cd. The binding of Cd to the fungal biomass was an extremely rapid process. During the first 5 minutes, 72.75% of Cd was taken up by the fungal biomass. The fungal Cd sorption was increased in low pH (range from 3 to 5).

[Test for the effects of mutagenic toxic fungal metabolites originating from municipal landfill sites]. / Badanie efektu mutagennego metabolitów grzybów toksynotwórczych pochodzacych z wysypiska odpadów komunalnych.

The purpose of the study is to assess the mutagenic effect of mycotoxins produced by moulds growing on municipal landfill sites. Mutagenicity of toxic fungal metabolites was determined by the Salmonella plate incorporation assay with two strains of bacteria: TA98 and TA100, with and without metabolic activation. The results obtained indicate that there is a severe hazard caused by these mycotoxins detected main by TA98 with metabolic activation. The most mutagenic mixture of mycotoxins acting directly on both strains was produced by Aspergillus nidulans. The highest mutagenic effect detected by TA98 with metabolic activation was found in the mixture of mycotoxins produced by one of three isolated Aspergillus fumigatus varieties.

First isolation of Microsporum racemosum Borelli in Poland.

During a study of keratinolytic fungi in the ash heap ground, considerably influenced by the municipal landfill site, a rare potentially pathogenic dermatophyte, Microsporum racemosum Borelli, was recognized. This is the first report on its isolation in Poland. Morphological and ecological features of the fungus are discussed.

Keratinolytic fungi in sewage sludge.

Sewage sludge from the Upper Silesia Region of Poland were surveyed for keratinolytic fungi. Out of 100 Petri dishes examined, 89 were positive for these micro-organisms. Altogether, 185 fungal appearances belonging to 10 species were observed. Trichophyton terrestre with its teleomorph Arthroderma quadrifidum, T. ajelloi with A. uncinatum, Microsporum gypseum with Arthroderma sp., and Chrysosporium keratinophilum with Aphanoascus keratinophilus prevailed in the sludges. The sewage treatment technologies together with the sludge structure, humidity and pH were found to be critical factors determining the occurrence of keratinolytic fungi in the sludge environment. The qualitative and quantitative composition of keratinolytic fungi could be a useful tool in evaluation of sludge treatment processes.

A statistical evaluation of the occurrence of keratinolytic fungi in the sediments of two dam reservoirs.

Sediments from two dam reservoirs "Przeczyce" and "Plawniowice" were examined for keratinolytic fungi. The results show the dependence of keratinolytic fungi in sediments on the degree of water contamination with sewage. Ch. keratinophilum is species associated with sewage inputs to superficial water.

Isolation of keratinolytic fungi from a coal mine dump.

During the study of fungal succesion in the coal mine dump in Brzezinka (Poland), soil samples were examined for keratinolytic fungi. These micro-organisms were rather poorly represented in the area studied. Out of 300 soil samples examined, only 48 (16%) were positive for keratinolytic fungi.Trichophyton ajelloi andArthroderma curreyi were the prevailing species. These species occurred practically at two locations, i.e. on the naked carbon rocks inhabited by algae crops (chiefly byCyanophyta) and in the pine litter. It can be supposed that the occurrence of keratinolytic fungi was more dependent on the favourable general conditions such as increasing organic matter content, microflora, and humidity than on the presence of keratin remains in the soil. Because of the lack of potentially pathogenic fungi, the coal mine dump examined cannot be considered as an important source of fungal infection.

[The occurrence of keratinolytic fungi in the polluted environment of the Labedy District in Gliwice]. / Wystepowanie grzybów keratynolitycznych w zanieczyszczonym srodowisku dzielnicy Gliwic-Labedy.

This study was undertaken to find relationships between the degree of bacteriological contamination with qualitative composition of potentially pathogenic keratinolytic fungal population in soil, sediment and air samples from the Labedy district in Gliwice (Poland). The examined soil samples were characterized by the predominance of Botryotrichum piluliferum, Chrysosporium anamorph of Arthroderma curreyi, Myceliophthora anamorph of Ctenomyces serratus, Chrysosporium pannicola and Trichphyton ajelloi. These species are typical for keratinolytic mycoflora in moderate climate soils, and their abundance was certainly resulted from the assembly of keratin remains in the soil environment. In the light of the weak bacteriological and mycological differentiation of the examined soil samples, however, it is difficult to determine categorically the extent to which the remains were originated from sewage via soil flooding by sewage or air transportation, or from the local human and animal population. Subsequently, the population of keratinolytic fungi in sediments was found to be clearly dependent of the degree of water contamination with sewage. In badly polluted sediments, Chrysosporium pannicola, Chrysosporium anamorph of Aphanoascus fulvescens, Chrysosporium keratinophilum, Trichophyton ajelloi and Microsporum cookei were prevalent species. Keratinolytic fungi were only a small part of airborne fungal population in sewage bioaerosoles. Geomyces pannorum, a soil species better known by its celullotytic than keratinolytic properties, predominated in air samples. Some pathogenic species, such as Aspergillus flavus, Aureobasidium pullulans, Chrysporium anamorph of Aphanoascus fulvescens, Candida spp., Geotrichum candidum, Microsporum canis, Sporothrix schenckii and Trichosporon beigelii, were recovered in the present study. This confirms that the sewage-contaminated environments in an important storage place of pathogenic fungi, mostly from the opportunistic subgroup.

[Keratinophilic fungi in waste water sediments]. / Grzyby keratynofilne w osadach sciekowych.

In 21 samples of various kinds of waste water sediments derived from 10 waste water treatment plants of Upper Silesia, the occurrence of keratinophilic fungi was examined. A 2-year recultivation cycle of one of the sediments was investigated. Keratinophilic fungi were found to occur abundantly in waste water sediments. The qualitative and quantitative composition of these fungi depends on the origin and composition of waste water sediments, stabilization degree of their organic matter, hydration degree and structure. It was postulated that keratinophilic fungi may be utilized as microbiological indices of the transformations of organic matter of waste water sediments as well as of the degree of their deactivation from the sanitary standpoint. These fungi can be particularly useful upon utilization of the sediments for recultivation and fertilization of grounds.

[Keratinophilic fungi in domestic gardens]. / Grzby keratynofilne w ogródkach przydomowych.

The quantitative compositions and growth of keratinophilic fungi in fertile and polluted soils of home-attached gardens in Upper Silesia were reported. The investigated soil samples were characterized by intensive growth of these fungi. In total, 49 species of microscopic fungi were isolated and identified. The main factors influencing the occurrence of keratinophilic fungi in soil were discussed.