Plant establishment, presence of arbuscular mycorrhizal fungi (AMF) and other rhizospheric fungi were studied in mine wastes from Zimapan, Hidalgo state, Mexico, using a holistic approach. Two long-term afforested and three non-afforested mine tailings were included in this research. Fifty-six plant species belonging to 29 families were successfully established on the afforested sites, while unmanaged tailings had only a few native plant species colonizing the surrounding soils. Almost all plant roots collected were associated to AMF in these sites. The genus Glomus was the most abundant AMF species found in their rhizosphere; however, the Acaulospora genus was also observed. Other rhizospheric fungi were identified by 18S rDNA sequencing analysis. Their role in these substrates, i.e. biocontrol, pollutant- and organic matter-degradation, and aides that increase plant metal tolerance is discussed. Our results advance the understanding of fungal diversity in sites polluted with metals and present alternative plants for remediation use.

A pot experiment was conducted to study the effects of high concentrations of available Cu and Pb in soil originated from the vicinity of a copper foundry in Poland (Cu, 2,585–3,725 mg kg−1 d.wt.; Pb, 1,459–1,812 mg kg−1 d.wt.) on the growth and chemical constituents of Betula pendula seedlings. Control plants grew in unpolluted forest soil. Dry matter accumulation in the plants during the growing season and root/leaf mineral content were determined. Colonization of birch roots by ectomycorrhizal (ECM) fungi also was evaluated, as was soil dehydrogenase activity for influence of the metals on soil microorganisms. The heavy metals negatively affected seedling growth, ECM colonization, and soil dehydrogenase activity. A reverse relationship was found between ECM abundance and heavy metal concentrations in birch leaves, indicating the potential of mycorrhizas to protect the aboveground part of young silver birch seedlings from elevated environmental levels of Cu and Pb.

Decolorization of six synthetic dyes and two raw textile effluents (A and B) by eight basidiomycetous fungi was investigated. Among eight basidiomycetous fungi, fungal isolate RCK-1 decolorized textile effluent A maximally (42%), while fungal isolate RCK-3 was found to decolorize more of Congo Red (69%), Xylidine Ponceau 2R (100%), Poly R-478 (96%), Indigo Carmine (99%), Lissamine Green B (90%), Toluidine Blue (57%) and textile effluent B (54%), than the rest of fungi. Percentage decolorization of all synthetic dyes and textile effluents by the new fungal isolates RCK-1 and RCK-3 was higher compared to the most widely studied simultaneous lignin degrader, Phanerochaete chrysosporium and selective lignin degrader, Pycnoporus cinnabarinus, when tested in liquid cultures. A statistically significant positive correlation between laccase production and decolorization of dyes and effluents was obtained as compared to other ligninolytic enzymes (lignin peroxidase and manganese peroxidase) production. This showed the importance of the differential contribution of the different ligninolytic enzymes towards the decolorization of the synthetic dyes and textile effluents. The substantially higher ligninolytic enzyme production by the fungal isolates RCK-1 and RCK-3 also suggested their potential use for textile effluent treatment and other possible biotechnological applications.

Investigations were made on living strains of fungi in a bioremediation process of three metal (lead) contaminated soils. Three saprotrophic fungi (Aspergillus niger, Penicillium bilaiae, and a Penicillium sp.) were exposed to poor and rich nutrient conditions (no carbon availability or 0.11 M d-glucose, respectively) and metal stress (25 µM lead or contaminated soils) for 5 days. Exudation of low molecular weight organic acids was investigated as a response to the metal and nutrient conditions. Main organic acids identified were oxalic acid (A. niger) and citric acid (P. bilaiae). Exudation rates of oxalate decreased in response to lead exposure, while exudation rates of citrate were less affected. Total production under poor nutrient conditions was low, except for A. niger, for which no significant difference was found between the poor and rich control. Maximum exudation rates were 20 µmol oxalic acid g⁻¹ biomass h⁻¹ (A. niger) and 20 µmol citric acid g⁻¹ biomass h⁻¹ (P. bilaiae), in the presence of the contaminated soil, but only 5 µmol organic acids g⁻¹ biomass h⁻¹, in total, for the Penicillium sp. There was a significant mobilization of metals from the soils in the carbon rich treatments and maximum release of Pb was 12% from the soils after 5 days. This was not sufficient to bring down the remaining concentration to the target level 300 mg kg⁻¹ from initial levels of 3,800, 1,600, and 370 mg kg⁻¹in the three soils. Target levels for Ni, Zn, and Cu, were 120, 500, and 200 mg kg⁻¹, respectively, and were prior to the bioremediation already below these concentrations (except for Cu Soil 1). However, maximum release of Ni, Zn, and Cu was 28%, 35%, and 90%, respectively. The release of metals was related to the production of chelating acids, but also to the pH-decrease. This illustrates the potential to use fungi exudates in bioremediation of contaminated soil. Nonetheless, the extent of the generation of organic acids is depending on several processes and mechanisms that need to be further investigated.

The concentration and size distribution of bacterial and fungal aerosol was studied in 15 houses. The houses were categorized into three types, based on occupant density and number of rooms: single room in shared accommodation (type I), single bedroom flat in three storey buildings (type II) and two or three bedroomed houses (type III). Sampling was undertaken with an Anderson six-stage impactor during the summer of 2007 in the living rooms of all the residential settings. The maximum mean geometric concentration of bacterial (5,036 CFU/m³, ± 2.5, n = 5) and fungal (2,124 CFU/m³, ± 1.38, n = 5) aerosol were in housing type III. The minimum levels of indoor culturable bacteria (1,557 CFU/m³, ±1.5, n = 5) and fungal (925 CFU/m³, ±2.9, n = 5) spores were observed in housing type I. The differences in terms of total bacterial and fungal concentration were less obvious between housing types I and II as compared to type III. With reference to size distribution, the dominant stages for culturable bacteria in housing types I, II and III were stage 3 (3.3-4.7 μm), stage 1 (7 μm and above) and stage 5 (1.1-2.1 μm), respectively. Whereas the maximum numbers of culturable fungal spores were recovered from stage 2 (4.7-7 µm), in housing type I, and from stage 4 (2.1-3.3 μm) in both type II and III houses. The average geometric mean diameter of bacterial aerosol was largest in type I (4.7 μm), followed by type II (3.89 μm) and III (1.96 μm). Similarly, for fungal spores, type I houses had the highest average mean geometric diameter (4.5 μm), while in types II and III the mean geometric diameter was 3.57 and 3.92 μm, respectively. The results indicate a wide variation in total concentration and size of bioaerosols among different residential settings. The observed differences in the size distributions and concentrations reflect their variable airborne behaviour and, as a result, different risks of respiratory exposure of the occupants to bioaerosols in various residential settings.

Exudation of low molecular weight organic acids by fungi was studied in a project focusing on bioremediation of metal-contaminated soils. The production of acids (mainly oxalic and citric acid) as a response to nutrient variations and presence of metals has recently been reported (Arwidsson et al. 2009). A significant release of metals was observed and was related not only to the production of organic acids but also to the resulting pH decrease in the systems. The processes governing the release and redistribution of metals in the soil–water fungus system were the focus of the present continuation of the project, based on observations of Aspergillus niger, Penicillium bilaiae, and a Penicillium sp. The release of lead was 12% from the soil with the second highest initial load (1,600 mg kg−1), while the release of copper was 90% from the same soil (140 mg kg−1). The dominating mechanism behind the release and subsequent redistribution was the change in pH, going from near neutral to values in the range 2.1–5.9, reflecting the production of organic acids. For some of the systems, the formation of soluble complexes is indicated (copper, at intermediate pH) which favors the metal release. Iron is assumed to play a key role since the amount of secondary iron in the soils is higher than the total load of secondary heavy metals. It can be assumed that most of the heavy metals are initially associated with iron-rich phases through adsorption or coprecipitation. These phases can be dissolved, or associated metals can be desorbed, by a decrease in pH. It would be feasible to further develop a process in technical scale for remediation of metal-contaminated soil, based on microbial metabolite production leading to formation of soluble metal complexes, notably with copper.

Bioaerosols are conglomerates of biological particles such as bacterial and fungal propagules and are produced in sewers and sewage treatment plants through evaporation and turbulence. In order to evaluate the hazard to employees in wastewater treatment plants, airborne microorganisms were measured at two different sites in the sewage systems and in the grit chamber of a treatment plant. Two additional samples were taken during high-pressure cleaning in the relief sewer. Outdoor air samples served as background values. Airborne microorganisms were collected using the impaction method with the MAS-100® and the impingement method with the SKC Biosampler®. The concentrations of coliform bacteria as well as the fungal species Aspergillus fumigatus were determined in addition to mesophilic bacteria counts (cfu/m³). The highest concentrations of mesophilic bacteria were found in the encased grit chamber. Coliform bacteria were found infrequently only in the aerosol of the sewage systems; A. fumigatus was detected at all sampling sites both indoors as well as outdoors. During high-pressure cleaning, total bacteria concentrations reached up to 4.0 × 104 cfu/m3, coliforms up to 3.0 × 103 cfu/m3. These results show that personnel protective measures should be recommended to decrease the exposure risk to biological particles.

Background, aim and scope Pulp and paper mills generate a plethora of pollutants depending upon the type of pulping process. Efforts to mitigate the environmental impact of such effluents have been made by developing more effective biological treatment systems in terms of biochemical oxygen demand, chemical oxygen demand, colour and lignin content. This study is the first that reports an evaluation of the effects of a tertiary treatment by fungi (Pleurotus sajor caju, Trametes versicolor and Phanerochaete chrysosporium and Rhizopus oryzae) on individual organic compounds of a Eucalyptus globulus bleached kraft pulp and paper mill final effluent after secondary treatment (final effluent). Material and methods The tertiary treatment with P. sajor caju, T. versicolor and P. chrysosporium and R. oryzae was performed in batch reactors, which were inoculated with separate fungi species and monitored throughout the incubation period. Samples from effluent after secondary and after tertiary treatment with fungi were analysed for both absorbance and organic compounds. The samples were extracted for organic compounds using solid-phase extraction (SPE) and analysed by gas chromatography-mass spectrometry (GC/MS). The efficiencies of the SPE procedure was evaluated by recovery tests. Results A total of 38 compounds (carboxylic acids, fatty alcohols, phenolic compounds and sterols) were identified and quantified in the E. globulus bleached kraft pulp mill final effluent after secondary treatment. Recoveries from the extraction procedure were between 98.2% and 99.9%. The four fungi species showed an adequate capacity to remove organic compounds and colour. Tertiary treatment with R. oryzae was able to remove 99% of organic compounds and to reduce absorbance on 47% (270 nm) and 74% (465 nm). P. sajor caju, T. versicolor and P. chrysosporium were able to remove 97%, 92% and 99% of organic compounds, respectively, and reduce 18% (270 nm) to 77% (465 nm), 39% (270 nm) to 58% (465 nm) and 31% (270 nm) to 10% (465 nm) of absorbance, respectively. Discussion The wide variety of organic compounds found in the final effluent must be due to the degradation of E. globulus wood in pulp and paper mill. The concentrations of organic compounds in the final effluent of E. globulus bleached kraft pulp mill were in residual levels maybe due to the secondary treatment. The recovery tests showed the effectiveness of the extraction procedure, and no losses of analyte were suspected in the analytical determinations. Lignin derivatives such as vanilic acid, syringic acid, guaiacol, syringol and phloroglucinol were totally removed by R. oryzae, but the 47% absorbance reduction obtained at 270 nm suggests that these species were not able to complete degradation of lignin macromolecular compounds. Conclusions The organic compounds (carboxylic acids, fatty alcohols, phenolic compounds and sterols) were removed more efficiently by tertiary treatment with R. oryzae or P. chrysosporium, followed by P. sajor caju and T. versicolor. Regarding the removal of both colour and organic compounds, the tertiary treatment with R. oryzae was the most efficient. Recommendations and perspectives In order to reduce the deleterious impacts of paper mill effluents, efforts have been made to develop more effective advanced tertiary treatments. This study may serve as a basis of characterisation, in terms of organic compounds of E. globulus bleached kraft pulp mill final effluent after secondary treatment and as an effort to understand the effects of tertiary treatments with fungi on low concentrations of organic compounds from biological secondary treatment.

Background, aim, and scope To stimulate the national economy, a so-called “gold week” comprising May Day and National Day has been put in force by the government, and the first golden-week holiday began on October 1, 1999. Statistical data show that about 15,000 visitors were received each day by Emperor Qin's Terra-Cotta Museum during just such a gold week period. To evaluate the effects of tourism on indoor air, airborne samples were collected by the sedimentation plate method for 5 min during the “Oct. 1” gold week period of 2006, and both composition and changes of airborne bacteria and fungi in indoor/outdoor air in the museums were investigated. Materials and methods Airborne microbes were simultaneously collected by means of gravitational sedimentation on open Petri dishes. Three parallel samples were collected at the same time each day, and samples were subsequently incubated in the lab. Microbiology media were prepared before each experiment by a professional laboratory. Concentrations were calculated and presented as average data of colony-forming units per cubic meter of air (CFU/m³). Results The results show that (1) 13 bacterial genera and eight genera of fungi were identified from indoor and outdoor air at Emperor Qin's Terra-Cotta Museum during “Oct. 1” gold week in 2006. The bacterial groups occupied 61%, the fungi groups occupied 36%, and others occupied 3% of the total number of isolated microorganism genera. (2) As for the comparison of indoor and outdoor samples, the average concentrations of fungi were higher during the afternoon (13:00) than for the morning (09:00). The average concentrations of bacteria in indoor air were higher during the afternoon (13:00) than for the morning (9:00), and in outdoor air, they were lower during the afternoon (13:00) than for the morning (9:00). (3) The average concentrations of five dominant groups of bacteria and three dominant groups of fungi were higher during the afternoon (13:00) than for the morning (9:00) in the indoor air, but the average concentrations of fungi were higher and those of bacteria were lower during the afternoon than for the morning, for outdoor air. (4) As for the comparison of indoor samples, the bacterial daily concentrations and fungal daily concentrations were higher during the afternoon (13:00) than those for the mornings (9:00) over the 10 days. For the comparison of outdoor samples, the bacterial concentration was lower, and the fungal concentrations were higher during the afternoon (13:00) than those for the morning (9:00) over the 10 days. Discussion The results also show that the numbers of airborne bacteria and fungi had a daily character in indoor air and were higher in the afternoon. The airborne microbe concentrations were found to be similar to residential indoor values from other reports; the indoor museum maximum of microbial concentrations was 90 CFU/m³ and did not exceed the Chinese indoor bioaerosol guideline. However, microorganisms may fall on the surface of display items as a result of particle sedimentation and would, as such, be capable of degrading objects by way of their secretions, e.g., enzymes and organic acids. Therefore, the right steps should be taken to prevent any deterioration in the quality of displayed artifacts. Conclusions The results show that museum air was affected by human activity; therefore, it is imperative that the number of visitors be strictly limited and that windows be opened regularly to avoid air pollution. Recommendations and perspectives The data provide a significant scientific basis for indoor air quality control and museum scientific management. It is recommended that the number of visitors be strictly limited.

Background, aim, and scope Soil contamination with heavy metals occurs as a result of both anthropogenic and natural activities. Heavy metals could have long-term hazardous impacts on the health of soil ecosystems and adverse influences on soil biological processes. Soil enzymatic activities are recognized as sensors towards any natural and anthropogenic disturbance occurring in the soil ecosystem. Similarly, microbial biomass carbon (MBC) is also considered as one of the important soil biological activities frequently influenced by heavy metal contamination. The polymerase chain reaction-denaturing gradient gel electrophoresis (DGGE) has recently been used to investigate changes in soil microbial community composition in response to environmental stresses. Soil microbial community structure and activities are difficult to elucidate using single monitoring approach; therefore, for a better insight and complete depiction of the soil microbial situation, different approaches need to be used. This study was conducted in a greenhouse for a period of 12 weeks to evaluate the changes in indigenous microbial community structure and activities in the soil amended with different application rates of Cd, Pb, and Cd/Pb mix. In a field environment, soil is contaminated with single or mixed heavy metals; so that, in this research, we used the selected metals in both single and mixed forms at different application rates and investigated their toxic effects on microbial community structure and activities, using soil enzyme assays, plate counting, and advanced molecular DGGE technique. Soil microbial activities, including acid phosphatase (ACP), urease (URE), and MBC, and microbial community structure were studied. Materials and methods A soil sample (0-20 cm) with an unknown history of heavy metal contamination was collected and amended with Cd, Pb, and Cd/Pb mix using the CdSO₄ and Pb(NO₃)₂ solutions at different application rates. The amended soils were incubated in the greenhouse at 25 ± 4°C and 60% water-holding capacity for 12 weeks. During the incubation period, samples were collected from each pot at 0, 2, 9, and 12 weeks for enzyme assays, MBC, numeration of microbes, and DNA extraction. Fumigation-extraction method was used to measure the MBC, while plate counting techniques were used to numerate viable heterotrophic bacteria, fungi, and actinomycetes. Soil DNAs were extracted from the samples and used for DGGE analysis. Results ACP, URE, and MBC activities of microbial community were significantly lower (p < 0.05) in the metal-amended samples than those in the control. The enzyme inhibition extent was obvious between different incubation periods and varied as the incubation proceeded, and the highest rate was detected in the samples after 2 weeks. However, the lowest values of ACP and URE activities (35.6% and 36.6% of the control, respectively) were found in the Cd₃/Pb₃-treated sample after 2 weeks. Similarly, MBC was strongly decreased in both Cd/Pb-amended samples and highest reduction (52.4%) was detected for Cd₃/Pb₃ treatment. The number of bacteria and actinomycetes were significantly decreased in the heavy metal-amended samples compared to the control, while fungal cells were not significantly different (from 2.3% to 23.87%). In this study, the DGGE profile indicated that the high dose of metal amendment caused a greater change in the number of bands. DGGE banding patterns confirmed that the addition of metals had a significant impact on microbial community structure. Discussion In soil ecosystem, heavy metals exhibit toxicological effects on soil microbes which may lead to the decrease of their numbers and activities. This study demonstrated that toxicological effects of heavy metals on soil microbial community structure and activities depend largely on the type and concentration of metal and incubation time. The inhibition extent varied widely among different incubation periods for these enzymes. Furthermore, the rapid inhibition in microbial activities such as ACP, URE, and MBC were observed in the 2 weeks, which should be related to the fact that the microbes were suddenly exposed to heavy metals. The increased inhibition of soil microbial activities is likely to be related to tolerance and adaptation of the microbial community, concentration of pollutants, and mechanisms of heavy metals. The DGGE profile has shown that the structure of the bacterial community changed in amended heavy metal samples. In this research, the microbial community structure was highly affected, consistent with the lower microbial activities in different levels of heavy metals. Furthermore, a great community change in this study, particularly at a high level of contamination, was probably a result of metal toxicity and also unavailability of nutrients because no nutrients were supplied during the whole incubation period. Conclusions The added concentrations of heavy metals have changed the soil microbial community structure and activities. The highest inhibitory effects on soil microbial activities were observed at 2 weeks of incubation. The bacteria were more sensitive than actinomycetes and fungi. The DGGE profile indicated that bacterial community structure was changed in the Cd/Pb-amended samples, particularly at high concentrations. Recommendations and perspectives The investigation of soil microbial community structure and activities together could give more reliable and accurate information about the toxic effects of heavy metals on soil health.