Rapid Microbial Community Changes During Initial Stages of Pine Litter Decomposition.

Plant litter decomposition is a process enabling biogeochemical cycles closing in ecosystems, and decomposition in forests constitutes the largest part of this process taking place in terrestrial biomes. Microbial communities during litter decomposition were studied mainly with low-throughput techniques not allowing detailed insight, particularly into coniferous litter, as it is more difficult to obtain high quality DNA required for analyses. Motivated by these problems, we analyzed archaeal, bacterial, and eukaryotic communities at three decomposition stages: fresh, 3- and 8-month-old litter by 16/18S rDNA pyrosequencing, aiming at detailed insight into early stages of pine litter decomposition. Archaea were absent from our libraries. Bacterial and eukaryotic diversity was greatest in 8-month-old litter and the same applied to bacterial and fungal rDNA content. Community structure was different at various stages of decomposition, and phyllospheric organisms (bacteria: Acetobacteraceae and Pseudomonadaceae members, fungi: Lophodermium, Phoma) were replaced by communities with metabolic capabilities adapted to the particular stage of decomposition. Sphingomonadaceae and Xanthomonadaceae and fungal genera Sistotrema, Ceuthospora, and Athelia were characteristic for 3-month-old samples, while 8-month-old ones were characterized by Bradyrhizobiaceae and nematodes (Plectus). We suggest that bacterial and eukaryotic decomposer communities change at different stages of pine litter decomposition in a way similar to that in broadleaf litter. Interactions between bacteria and eukaryotes appear to be one of the key drivers of microbial community structure.

Resilience of Soil Microbial Communities to Metals and Additional Stressors: DNA-Based Approaches for Assessing "Stress-on-Stress" Responses.

Many microbial ecology studies have demonstrated profound changes in community composition caused by environmental pollution, as well as adaptation processes allowing survival of microbes in polluted ecosystems. Soil microbial communities in polluted areas with a long-term history of contamination have been shown to maintain their function by developing metal-tolerance mechanisms. In the present work, we review recent experiments, with specific emphasis on studies that have been conducted in polluted areas with a long-term history of contamination that also applied DNA-based approaches. We evaluate how the "costs" of adaptation to metals affect the responses of metal-tolerant communities to other stress factors ("stress-on-stress"). We discuss recent studies on the stability of microbial communities, in terms of resistance and resilience to additional stressors, focusing on metal pollution as the initial stress, and discuss possible factors influencing the functional and structural stability of microbial communities towards secondary stressors. There is increasing evidence that the history of environmental conditions and disturbance regimes play central roles in responses of microbial communities towards secondary stressors.

Soil chemical properties affect the reaction of forest soil bacteria to drought and rewetting stress.

Reaction of soil bacteria to drought and rewetting stress may depend on soil chemical properties. The objectives of this study were to test the reaction of different bacterial phyla to drought and rewetting stress and to assess the influence of different soil chemical properties on the reaction of soil bacteria to this kind of stress. The soil samples were taken at ten forest sites and measured for pH and the contents of organic C (Corg) and total N (Nt), Zn, Cu, and Pb. The samples were kept without water addition at 20 - 30 °C for 8 weeks and subsequently rewetted to achieve moisture equal to 50 - 60 % of their maximum water-holding capacity. Prior to the drought period and 24 h after the rewetting, the structure of soil bacterial communities was determined using pyrosequencing of 16S rRNA genes. The drought and rewetting stress altered bacterial community structure. Gram-positive bacterial phyla, Actinobacteria and Firmicutes, increased in relative proportion after the stress, whereas the Gram-negative bacteria in most cases decreased. The largest decrease in relative abundance was for Gammaproteobacteria and Bacteroidetes. For several phyla the reaction to drought and rewetting stress depended on the chemical properties of soils. Soil pH was the most important soil property influencing the reaction of a number of soil bacterial groups (including all classes of Proteobacteria, Bacteroidetes, Acidobacteria, and others) to drought and rewetting stress. For several bacterial phyla the reaction to the stress depended also on the contents of Nt and Corg in soil. The effect of heavy metal pollution was also noticeable, although weaker compared to other chemical soil properties. We conclude that soil chemical properties should be considered when assessing the effect of stressing factors on soil bacterial communities.

Metals affect soil bacterial and fungal functional diversity differently.

Heavy metals can cause a decrease in the taxonomic diversity of soil communities. Because of functional redundancy, it remains unclear to what extent different functions performed by the soil microbial communities may be affected by pollution. We studied the impact of metal contamination on soil bacterial and fungal functional diversity, active microbial biomass, and soil respiration rate. Soil samples were collected from 39 sites along three forest and five meadow pollution transects near an abandoned Pb/Zn smelter in Avonmouth (UK) and Ni smelter in Clydach (UK), in a Cu mining and smelting region near Glogów (Poland), and in a Zn/Pb mining and smelting region near Olkusz (Poland). Biolog GN2 and SFN2 plates were used to study the bacterial and fungal functional diversity, which subsequently was expressed as Shannon's diversity index (H'). The active microbial biomass was measured as substrate-induced respiration. We found that the bacterial functional diversity significantly decreased, whereas the fungal functional diversity slightly increased, with increasing metal concentration. We also observed a slight negative effect of metal pollution on the active microbial biomass. No relationship was found between metal contamination and total soil respiration rate. This suggests a higher sensitivity of bacterial functional diversity as an indicator for the effects of metal pollution compared with overall soil respiration. All microbial parameters were affected by nutrient concentrations and/or soil pH.

The Effects of Heavy Metals and Total Petroleum Hydrocarbons on Soil Bacterial Activity and Functional Diversity in the Upper Silesia Industrial Region (Poland).

Various inorganic and organic pollutants in industrial soils may adversely affect soil microorganisms and terrestrial ecosystem functioning. The aim of the study was to explore the relationship between the microbial activity, microbial biomass, and functional diversity of soil bacteria and the metals and total petroleum hydrocarbons (TPHs) in the Upper Silesian Industrial Region (Poland). We collected soil samples in pine-dominated forest stands and analyzed them according to a range of soil physicochemical properties, including metal content (cadmium, lead, and zinc) and TPH content. Metal concentrations were normalized to their toxicity to soil microorganisms and integrated in a toxicity index (TI). Soil microbial activity measured as soil respiration rate, microbial biomass measured as substrate-induced respiration rate, and the bacterial catabolic activity (area under the curve, AUC) assessed using Biolog® ECO plates were negatively related to TPH pollution as shown in multiple regressions. The canonical correspondence analysis (CCA) showed that both TPH and TI affected the community-level physiological profiles (CLPPs) of soil bacteria and the pollutants' effects were much stronger than the effects of other soil properties, including nutrient content.

Elemental changes in the brain, muscle, and gut cells of the housefly, Musca domestica, exposed to heavy metals.

The toxic effects of heavy metals on organisms are well established. However, their specific action at the cellular level in different tissues is mostly unknown. We have used the housefly, Musca domestica, as a model organism to study the toxicity of four heavy metals: copper (Cu), zinc (Zn), cadmium (Cd), and lead (Pb). These have been fed to larvae at low and high, semi-lethal concentrations, and their accumulation in the head, thorax, and abdomen was subsequently measured in adult flies. In addition, their impact on the cellular concentration of several elements important for cell metabolism-sodium (Na+), magnesium (Mg++), phosphorous (P), sulphur (S), chloride (Cl-) and potassium (K+)-were measured in neural cells, muscle fibers, and midgut epithelial cells. Our study showed that the heavy metals accumulate mainly in the abdomen, in which the concentrations of two of the xenobiotic metals, Cd and Pb, were 213 and 23 times more concentrated, respectively, than in controls. All the heavy metals affected the cellular concentration of light elements in all cell types, but the changes observed were dependent on tissue type and were specific for each heavy metal, and its concentration.

Microbial community composition and functions are resilient to metal pollution along two forest soil gradients.

Despite the global importance of forests, it is virtually unknown how their soil microbial communities adapt at the phylogenetic and functional level to long-term metal pollution. Studying 12 sites located along two distinct gradients of metal pollution in Southern Poland revealed that functional potential and diversity (assessed using GeoChip 4.2) were highly similar across the gradients despite drastically diverging metal contamination levels. Metal pollution level did, however, significantly impact bacterial community structure (as shown by MiSeq Illumina sequencing of 16S rRNA genes), but not bacterial taxon richness and community composition. Metal pollution caused changes in the relative abundance of specific bacterial taxa, including Acidobacteria, Actinobacteria, Bacteroidetes, Chloroflexi, Firmicutes, Planctomycetes and Proteobacteria. Also, a group of metal-resistance genes showed significant correlations with metal concentrations in soil. Our study showed that microbial communities are resilient to metal pollution; despite differences in community structure, no clear impact of metal pollution levels on overall functional diversity was observed. While screens of phylogenetic marker genes, such as 16S rRNA genes, provide only limited insight into resilience mechanisms, analysis of specific functional genes, e.g. involved in metal resistance, appears to be a more promising strategy.

Effects of cadmium and zinc on larval growth and survival in the ground beetle, Pterostichus oblongopunctatus.

Carabid beetles, like Pterostichus oblongopunctatus, living in metal contaminated areas may be exposed to elevated levels of metals within their diets. However, when compared to other second order consumers, they have one of the lowest observed levels of metals, indicating methods of detoxification to deal with such toxicants. In this study, we investigated if chronic, multigenerational exposure to metals leads to resistance to toxic metal concentrations, and if so, what are the costs associated with them. Adult organisms were collected from two sites, a polluted and a reference site near Olkusz, in southern Poland. These adults were immediately mated, and eggs were collected twice weekly to assess the effects in the larvae of the F(1) generation. Larvae were randomly exposed to one of four artificial mediums: control, 50 mg kg(-1) Cd, 500 mg kg(-1) Zn, and a combined treatment of 50 mg kg(-1) Cd and 500 mg kg(-1) Zn to investigate possible interactions. Individuals were sacrificed at 10, 30, and 40 days. Although metals were not accumulated in larvae (p>0.05), larvae fed the Cd or the Zn treatment grew significantly slower, and had the lowest survival rate (p<0.05) in respect to control. Out of metal treated animals, those on the combined treatment of CdZn grew the quickest and had the highest observed survival (p<0.05). Although previous studies have demonstrated changes in adult population parameters under chronic, multigeneration exposure to toxic metal concentrations, our study did not reveal any changes in the larval stage.

Microbial community structure and functioning along metal pollution gradients.

Toxic effects of heavy metals on soil microorganisms have been confirmed in a number of laboratory studies. However, most real-field studies do not allow for strong general conclusions due to a range of problems, such as pseudoreplication and confounding factors, which are almost impossible to control for with the most commonly used polluted versus unpolluted or random sampling designs. Effects of metal contamination on soil microbial community traits were measured along 2 pollution gradients in southern Poland. Employing an experimental regression design, using 2 separate gradients, the authors aimed to control for effects of soil properties and beta-diversity of microbial communities. General microbial activity was measured as soil basal respiration rate and substrate-induced respiration, while microbial functional and structural diversity were analyzed with community-level physiological profiles and phospholipid fatty acid patterns, respectively. Metal concentrations were normalized to their toxicity and integrated in a toxicity index (TI). Microbial activity (basal and substrate-induced respiration) decreased in both gradients with increasing TI. Community-level physiological profiles for fungi correlated positively with TI, but no impact of TI on the community-level physiological profiles of bacteria was observed. The phospholipid fatty acids a:15 and i:17 were positively correlated with TI, whereas 16:1ω9 and 18:2ω9 were negatively correlated with TI. The use of 2 gradients (Olkusz and Miasteczko Slaskie) allowed the authors to reveal a clear effect of pollution on general microbial structure and activities, even though they were not able to control completely for all confounding factors. Soil pH, organic matter content, and nutrient level appeared to be at least as important as TI in determining microbial community structure and activities.

Soil fertility and plant diversity enhance microbial performance in metal-polluted soils.

This study examined the effects of soil physicochemical properties (including heavy metal pollution) and vegetation parameters on soil basal respiration, microbial biomass, and the activity and functional richness of culturable soil bacteria and fungi. In a zinc and lead mining area (S Poland), 49 sites were selected to represent all common plant communities and comprise the area's diverse soil types. Numerous variables describing habitat properties were reduced by PCA to 7 independent factors, mainly representing subsoil type (metal-rich mining waste vs. sand), soil fertility (exchangeable Ca, Mg and K, total C and N, organic C), plant species richness, phosphorus content, water-soluble heavy metals (Zn, Cd and Pb), clay content and plant functional diversity (based on graminoids, legumes and non-leguminous forbs). Multiple regression analysis including these factors explained much of the variation in most microbial parameters; in the case of microbial respiration and biomass, it was 86% and 71%, respectively. The activity of soil microbes was positively affected mainly by soil fertility and, apparently, by the presence of mining waste in the subsoil. The mining waste contained vast amounts of trace metals (total Zn, Cd and Pb), but it promoted microbial performance due to its inherently high content of macronutrients (total Ca, Mg, K and C). Plant species richness had a relatively strong positive effect on all microbial parameters, except for the fungal component. In contrast, plant functional diversity was practically negligible in its effect on microbes. Other explanatory variables had only a minor positive effect (clay content) or no significant influence (phosphorus content) on microbial communities. The main conclusion from this study is that high nutrient availability and plant species richness positively affected the soil microbes and that this apparently counteracted the toxic effects of metal contamination.

Pine forest and grassland differently influence the response of soil microbial communities to metal contamination.

Metal pollution can affect soil microbial communities, and vegetation potentially influences this relationship. It can, for example, modify the toxicity of metal to soil microbes by controlling its input to the ground or by altering soil physicochemical properties. This study examined metal effects on soil respiration, potentially active microbial biomass (SIR) and catabolic abilities of culturable heterotrophic bacterial communities (Biolog GN) in pine forest and grassland ecosystems developed on soils contaminated with Zn, Pb and Cd. In samples from non-forested areas we found that metal pollution reduced the microbial biomass and functional diversity of bacteria, while increasing the metabolic quotient. In samples from pine forests we found no relationship between metal pollution and microbial parameters. Metals induced changes in soil respiration neither in forest nor in grassland sites. Generally, microbial performance was determined predominantly by soil physicochemical properties (nutrient content, acidity, contamination level). Vegetation type seemed a minor but important factor influencing microbial communities. More work is needed to determine why even relatively high metal concentrations do not significantly affect microbial communities in forest soils.

Decreased energetic reserves, morphological changes and accumulation of metals in carabid beetles (Poecilus cupreus L.) exposed to zinc- or cadmium-contaminated food.

The prime objective of the study was to find out whether contamination of food with metals affects body size, energetic reserves and developmental instability in ground beetles (Poecilus cupreus L.: Carabidae). The transfer of Cadmium (Cd) and Zinc (Zn) from medium (nominal concentrations in the medium: 0, 40, 160, 640 or 800 mg kg(-1) for Cd and 0, 400, 1600 or 6400 mg kg(-1) for Zn) to housefly larvae to beetles was also studied. Feeding the beetles throughout their entire lifetime with Cd-contaminated housefly pupae resulted in a significant decrease in body caloric value and the size of the elytrae, tibiae and rear femora. Although body mass also decreased with increasing Cd concentration, this effect was non-significant due to large variance in all treatments. Similar trends were also found in beetles fed pupae contaminated with Zn, but the effect on body mass and caloric value was non-significant. Zn exerted significant effects only on the size of the elytrae, middle and rear tibiae, and front and rear femora. No effect on fluctuating asymmetry (FA) was found in Cd- or Zn-treated beetles. The results indicate that ground beetles exposed to metal-contaminated food have lower amounts of energy available, which may be reflected in lower energetic reserves and changed body growth. However, the metals do not cause developmental instability in the carabids studied, at least not in the first generation. The concentrations of Zn were efficiently regulated in carabids, resulting in only minor differences between the beetles exposed to different Zn treatments. In contrast, Cd accumulated both in the housefly and the beetles, and the concentrations increased significantly with increasing medium contamination level.