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The Brazilian National Environmental Council (CONAMA) Resolution 420/2009 establishes soil quality guideline values for copper, as the prevention value (PV). It defines the maximum concentrations of chemical substances permitted in the soil. Thus, the aim of the present research was to evaluate the effectiveness of the copper PV adopted at the national level by conducting exposure assays with enchytraeids (Enchytraeus crypticus, Enchytraeus bigeminus and Enchytraeus dudichi) and springtails (Folsomia candida, Proisotoma minuta and Sinella curviseta) in two Brazilian subtropical soils: a Latosol and an Argisol. Results revealed that copper reduces the reproductive capacity of all organisms as its concentration increases in the evaluated soils. Argisol presented the lowest effective concentration of 20% (EC 20 ) and 50% (EC 50 ) in species reproduction for all organisms evaluated. The most sensitive organism was the enchytraeid E. bigeminus, with EC 20 values of 58 mg kg⁻ 1 in Latosol and 30 mg kg⁻ 1 in Argisol, and EC 50 values of 155 mg kg⁻ 1 in Latosol and 91 mg kg⁻ 1 in Argisol. Among the springtails, S. curviseta exhibited the highest EC 20 , with values of 230 mg kg⁻ 1 in Latosol and 136 mg kg⁻ 1 in Argisol. Conversely, the highest values of EC 50 were recorded for the species P. minuta (526 mg kg⁻ 1 in Latosol and 415 mg kg⁻ 1 in Argisol). Based on the species sensitivity distribution (SSD) approach, the PVs calculated were 172 mg kg⁻ 1 for Latosol and 106 mg kg⁻ 1 for Argisol, derived from EC 50 data. For EC 20 data, the PV values calculated were 158 mg kg⁻ 1 in Latosol and 100 mg kg⁻ 1 in Argisol. By comparing the PV with the Brazilian adopted PV for copper (60 mg kg -1 ), it was observed the efficacy of the CONAMA Resolution., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier Ltd. All rights reserved.)

Copper is a trace element in plants and animals whose importance can be understood due to its role in different essential metabolic processes. Anthropogenic activities such as agriculture and mining are potential sources of pollution due to the emission of copper into the environment. Brazilian legislation ties soil quality to guideline values, among which the Prevention Value indicates the critical environmental limit for trace elements. The aim of this study was to obtain PVs for copper for two subtropical soils (Cambisol and Nitisol), given that the pedological richness was not considered when deriving the PVs contained in the federal normative. Reproduction assays followed ISO guidelines with the earthworm species Eisenia andrei and Perionyx excavatus, the enchytraeids Enchytraeus crypticus and Enchytraeus bigeminus and the springtails Folsomia candida and Proisotoma minuta. Results showed that the sensitivity of the organisms was greater in Cambisol. The most sensitive species were the earthworms, especially Perionyx excavatus (EC 50  = 67.83 in Cambisol; EC 50  = 264.96 in Nitisol). The springtails, on the other hand, were the least sensitive to contamination. These findings reinforce the need to include organisms from different ecological groups in ecotoxicological assessments. It was also observed that the PV adopted in federal legislation (= 60 mg kg -1 ) is in fact protective for the species and soils we evaluated, since the PVs we obtained based on the EC 50 were 346.74 mg kg -1 in Nitisol and 134.05 mg kg -1 in Cambisol. It is important to note that our results do not exclude the need for evaluations with other subtropical soils, given the influence of their properties on the toxicity and bioavailability of copper to soil organisms., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The increased production and expanding demand for cellulosic products have required companies to provide solutions for the proper disposal of generated residues without compromising soil quality. In this context, the current research aimed at evaluating the ecotoxicological impacts of dregs application in subtropical soils. The experiments were carried out with the earthworm Eisenia andrei, the springtail Folsomia candida, and the potworm Enchytraeus crypticus in two subtropical soils (Neosol and Cambisol) collected in the southern region of Brazil. The considerable differences in texture and organic matter content motivated the choosing of these soils. The reproductive rate of all organisms was influenced, with varying EC 50 values depending on the soil type. However, more pronounced effects were observed for Neosol. The springtail F. candida proved to be the most sensitive to contamination (EC 50  = 5.8 g kg -1 for Neosol and 48.5 g kg -1 for Cambisol), followed by the earthworm E. andrei (EC 50  = 62.4 g kg -1 for Neosol and 129.5 g kg -1 for Cambisol) and the potworm E. crypticus (EC 50  = 67.2 g kg -1 for Neosol and 230.4 g kg -1 for Cambisol). As a result, while dregs can be used to correct soil acidity, they have been shown to have negative effects on important organisms responsible for functions critical to maintaining ecosystem quality., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

In the last decade, advances in soil bacterial ecology have contributed to increasing agricultural production. Brazil is the world leading agriculture producer and leading soil biodiversity reservoir. Meanwhile, there is still a significant gap in the knowledge regarding the soil microscopic life and its interactions with agricultural practices, and the replacement of natural vegetation by agroecosystems is yet to be unfolded. Through high throughput DNA sequencing, scientists are now exploring the complexity of soil bacterial communities and their relationship with soil and environmental characteristics. This study aimed to investigate the progress of bacterial ecology studies in Brazil over the last 10 years, seeking to understand the effect of the conversion of natural vegetation in agricultural systems on the diversity and structure of the soil microbial communities. We conducted a systematic search for scientific publication databases. Our systematic search has matched 62 scientific articles from three different databases. Most of the studies were placed in southeastern and northern Brazil, with no records of studies about microbial ecology in 17 out of 27 Brazilian states. Out of the 26 studies that examined the effects of replacing natural vegetation with agroecosystems, most authors concluded that changes in soil pH and vegetation cover replacement were the primary drivers of shifts in microbial communities. Understanding the ecology of the bacteria inhabiting Brazilian soils in agroecosystems is paramount for developing more efficient soil management strategies and cleaner agricultural technologies., Competing Interests: The authors declare no conflict of interest.

Springtails (Collembola) inhabit soils from the Arctic to the Antarctic and comprise an estimated ~32% of all terrestrial arthropods on Earth. Here, we present a global, spatially-explicit database on springtail communities that includes 249,912 occurrences from 44,999 samples and 2,990 sites. These data are mainly raw sample-level records at the species level collected predominantly from private archives of the authors that were quality-controlled and taxonomically-standardised. Despite covering all continents, most of the sample-level data come from the European continent (82.5% of all samples) and represent four habitats: woodlands (57.4%), grasslands (14.0%), agrosystems (13.7%) and scrublands (9.0%). We included sampling by soil layers, and across seasons and years, representing temporal and spatial within-site variation in springtail communities. We also provided data use and sharing guidelines and R code to facilitate the use of the database by other researchers. This data paper describes a static version of the database at the publication date, but the database will be further expanded to include underrepresented regions and linked with trait data., (© 2024. The Author(s).)

Arbuscular mycorrhizal fungi (AMF) are a key group of soil organisms involved in several ecosystem services, but they had not been explored in retrospective ecological risk assessment (ERA) schemes, partially due to a lack of experimental protocols. We aimed to perform a site-specific test to assess the effects of multiple metal contaminated soil (Zn, Cu, Cd, and Pb) on several ecotypes of AMF and evaluate the conceptual fitness of the performed AMF test to include in the TRIAD approach of ERA schemes. The results demonstrated that increasing metal contaminated soil proportions that inhibited 50 % (IC 50 ) of spore germination varied from 28 to >80. Ecotypes such as Claroideoglomus etunicatum (IC 50  > 80) and Racocetra gregaria (IC 50  > 80) experienced 50 % reduction in spore germination at metals concentrations of 10,776.3 for Zn, 1015.2 Cu, 65.5 Cd, 140.2 mg dm -3 Pb, that are 3 times higher than those for Acaulospora mellea CMM101 (IC 50 28 [16.2-39.8]) (3441.7 Zn, 333.9 Cu, 17.8 Cd, 56.5 mg dm -3 Pb). In the evaluation of the suitability of the AMF ecotoxicological test to ERA, both spore germination and germinative tube growth were best evaluated and thus suitable in the following descending order: Tier III, Tier II, and Tier I. Variable effects of multiple-metal contamination on the ecotypes indicates how AMF community is affected in its pre-symbiotic structures. The ecotoxicological test allowed the selection of two species with the greatest sensitivity (Ambispora appendicula and Rhizophagus clarus CMM103) to the metal matrix, with the potential to best fit ERA objectives. The site-specific ecotoxicological test with AMF ex-situ proved adequate as an alternative test for Tiers II and III of TRIAD ERA schemes for metal contaminated areas. Data generated through test results, such as the inhibition concentrations (ICs), could be incorporated into ERAs risk indexes, increasing its ecological relevance, and reducing overall uncertainties., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Soil life supports the functioning and biodiversity of terrestrial ecosystems. Springtails (Collembola) are among the most abundant soil arthropods regulating soil fertility and flow of energy through above- and belowground food webs. However, the global distribution of springtail diversity and density, and how these relate to energy fluxes remains unknown. Here, using a global dataset representing 2470 sites, we estimate the total soil springtail biomass at 27.5 megatons carbon, which is threefold higher than wild terrestrial vertebrates, and record peak densities up to 2 million individuals per square meter in the tundra. Despite a 20-fold biomass difference between the tundra and the tropics, springtail energy use (community metabolism) remains similar across the latitudinal gradient, owing to the changes in temperature with latitude. Neither springtail density nor community metabolism is predicted by local species richness, which is high in the tropics, but comparably high in some temperate forests and even tundra. Changes in springtail activity may emerge from latitudinal gradients in temperature, predation and resource limitation in soil communities. Contrasting relationships of biomass, diversity and activity of springtail communities with temperature suggest that climate warming will alter fundamental soil biodiversity metrics in different directions, potentially restructuring terrestrial food webs and affecting soil functioning., (© 2023. The Author(s).)

Brazil is considered a megadiverse country, but the soil fauna is still very poorly known. The aim of this study was to report, for the first time, the abundance and genus composition of terrestrial enchytraeids (Enchytraeidae, Oligochaeta) in Savanna Tall Woodland (Cerradão) and a pasture in Cerrado Biome and in Upper Montane Atlantic Forest and a grassland in Atlantic Forest Biome. The enchytraeid density in Pasture and Cerradao was 2,036 and 18,844 (204 and 2,094, on average) individuals per square meter, respectively. At the Atlantic forest and Grassland, density was 9,666 and 12,242 individuals per square meter (1,075 and 1,471 on average). About genus composition for the studied areas, Enchytraeus and Hemienchytraeus were found in the four ecosystems evaluated, while Tupidrilus and Fridericia were found only in Cerradão and Atlantic Forest, respectively. Achaeta was absent in Upper Montane Atlantic Forest, but dominant in pasture, while Guaranidrilus was absent in Pasture, but predominant in the other ecosystems.

Understanding the effects of forest-to-agriculture conversion on microbial diversity has been a major goal in soil ecological studies. However, linking community assembly to the ruling ecological processes at local and regional scales remains challenging. Here, we evaluated bacterial community assembly patterns and the ecological processes governing niche specialization in a gradient of geography, seasonality, and land-use change, totaling 324 soil samples, 43 habitat characteristics (abiotic factors), and 16 metabolic and co-occurrence patterns (biotic factors), in the Brazilian Atlantic Rainforest, a subtropical biome recognized as one the world's largest and most threatened hotspots of biodiversity. Pairwise beta diversities were lower in pastures than in forest and no-till soils. Pasture communities showed a predominantly neutral model, regarding stochastic processes, with moderate dispersion, leading to biotic homogenization. Most no-till and forest microbial communities followed a niche-based model, with low rates of dispersal and weak homogenizing selection, indicating niche specialization or variable selection. Historical and evolutionary contingencies, as represented by soil type, season, and dispersal limitation were the main drivers of microbial assembly and processes at the local scale, markedly correlated with the occurrence of endemic microbes. Our results indicate that the patterns of assembly and their governing processes are dependent on the niche occupancy of the taxa evaluated (generalists or specialists). They are also more correlated with historical and evolutionary contingencies and the interactions among taxa (i.e., co-occurrence patterns) than the land-use change itself., (© 2022. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

This study aimed to evaluate the effects of untreated pig manure from diets incorporating growth-promoting supplements (antibiotics and Zn oxide) on the survival and reproduction of Eisenia andrei earthworms. The tested manures were obtained from four different groups of pigs fed with four different diets: CS, a diet based on corn and soymeal; TR, a diet based on corn, soymeal, and ground wheat (15%); CSa, a diet based on corn and soymeal + 100 ppm of doxycycline + 50 ppm of colistin + 2500 ppm of Zn oxide; and TRa, a diet based on corn, soymeal, and ground wheat (15%) + 100 ppm of doxycycline + 50 ppm of colistin + 2500 ppm of Zn oxide. The study used two soils representative of the Southern region of Brazil (Oxisol and Entisol). In general, there were no significant differences between the different manures tested in each soil. However, there were differences in the toxicity manure on E. andrei between the soils, and the magnitude of this effect was dependent on the applied dose. In Oxisol, LC 50 values were higher than 80 m 3  ha -1 , and EC 50 varied from 9 to 27 m 3  ha -1 . In Entisol, the LC 50 values were below the lowest dose tested (< 25 m 3  ha -1 ), and EC 50 remained around 5 m 3  ha -1 . It may be possible that the effects observed were attributed to an excess of nitrogen, copper, and zinc, promoted by the addition of the untreated manure and how these factors interacted with soil type., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Information concerning arbuscular mycorrhizal (AM) fungal geographical distribution in tropical and subtropical soils from the Atlantic Forest (a global hotspot of biodiversity) are scarce and often restricted to the evaluation of richness and abundance of AM fungal species at specific ecosystems or local landscapes. In this study, we hypothesized that AM fungal diversity and community composition in subtropical soils would display fundamental differences in their geographical patterns, shaped by spatial distance and land-use change, at local and regional scales. AM fungal community composition was examined by spore-based taxonomic analysis, using soil trap cultures. Acaulospora koskei and Glomus were found as generalists, regardless of mesoregions and land uses. Other Acaulospora species were also found generalists within mesoregions. Land-use change and intensification did not influence AM fungal composition, partially rejecting our first hypothesis. We then calculated the distance-decay of similarities among pairs of AM fungal communities and the distance-decay relationship within and over mesoregions. We also performed the Mantel test and redundancy analysis to discriminate the main environmental drivers of AM fungal diversity and composition turnover. Overall, we found significant distance-decays for all land uses. We also observed a distance-decay relationship within the mesoregion scale (< 104 km) and these changes were correlated mainly to soil type (not land use), with the secondary influence of both total organic carbon and clay contents. AM fungal species distribution presented significant distance-decays, regardless of land uses, which was indicative of dispersal limitation, a stochastic neutral process. Although, we found evidence that, coupled with dispersal limitation, niche differentiation also played a role in structuring AM fungal communities, driven by long-term historical contingencies, as represented by soil type, resulting from different soil origin and mineralogy across mesoregions., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC part of Springer Nature.)

Given the essential role of arbuscular mycorrhizal fungi (AMF) in soil systems and agriculture, their use as biological indicators has risen in all fields of microbiology research. However, AMF sensitivity to chemical pesticides is poorly understood in field conditions, and not explored in ecotoxicology protocols. Hence, the goal of this study was to evaluate the effects of different concentrations of glyphosate (Roundup®) and diuron+paraquat (Gramocil®) on the germination of spores of Gigaspora albida and Rhizophagus clarus in a tropical artificial soil. This study was conducted in 2019 at the Soil Ecology and Ecotoxicology Laboratory of the Universidade do Estado de Santa Catarina. The nominal concentrations of glyphosate were 0, 10, 50, 100, 250, 500, 750 and 1000 mg a.i. kg -1 . For diuron+paraquat, the concentrations tested were 0, 10 + 20, 50 + 100, 100 + 200, 250 + 500, 500 + 1000, 750 + 1500 and 1000 + 2000 mg a.i. kg -1 . Glyphosate did not alter germination of G. albida, but germination inhibition of R. clarus spores was of 30.8% at 1000 mg kg -1 . Diuron+paraquat inhibited by 8.0% germination of G. albida, but only at the highest concentration tested. On the other hand, effects on R. clarus were detected at 50 + 100 mg kg -1 concentration and above, and inhibition was as high as 57.7% at the highest concentration evaluated. These results suggest distinct response mechanisms of Rhizophagus and Gigaspora when exposed to herbicides, with the former being more sensitive than the later., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

The biobed is a purification system, which reduces soil pollution for receiving pesticide residues from handling and washing machinery in agricultural areas. The aims of this study were (1) to assess ecotoxicity effects over time to soil fauna, posed by Lorsban ® 480 BR (Chlorpyrifos) and Dithane ® NT (Mancozeb) residues when disposed of in a biobed system compared with two subtropical soils, and (2) to assess ecotoxicity effects over time to soil fauna simulating an accidental spillage with Lorsban ® 480 BR at the biobed. A semi-field experiment was conducted for 420 days in southern Brazil, testing continuous disposal of washing pulverization tanks in biobeds, Typic Haploperox or Typic Hapludults. In addition, different biobeds received a single dose (1 L) of Lorsban ® 480 BR to simulate an accidental spillage. Chronic ecotoxicity tests were performed using Folsomia candida, Eisenia andrei, and Enchytraeus crypticus in different sampling times for both experiments. F. candida was the most sensitive species. The biobed system was able to eliminate effects from residues of both pesticides over time in all species, which did not happen in both natural soils. In accidental spillage simulation, even 420 days after contamination, F. candida did not show reproduction. The biobeds can be a feasible alternative for the disposal and treatment residues of pesticides, also for handling and washing pesticides activities. The system was efficient in promoting degradation and reducing ecotoxicity effects posed by Lorsban ® 480 BR and Dithane ® NT for soil fauna. It is a safe alternative to avoid soil contamination.

Wastes generated in pig production are widely used as agricultural fertilizers. Nevertheless, such form of disposal supplies large amounts of waste in soils annually and can cause environmental pollution. The ecological risk of this practice to soil organisms has received little attention. Ecotoxicological tests are used to evaluate the toxicity of contaminants added to the soil biota. The aimed to evaluate the effect of liquid pig manure (LPM) on the reproduction of Eisenia andrei and Enchytraeus crypticus when applied in natural soils. LPM doses caused effects on earthworm reproduction in both soils, with EC 50 of 112 and 150 m 3  ha -1 in the Entisol and Nitosol, respectively. On enchytraeids, LPM had bigger effects, leading to EC 50 of 17.7 and 45.0 m 3  ha -1 in the Entisol and Nitosol, respectively. The results emphasize the importance to consider the ecological risks of LPM of conducting studies with natural soils and edaphic fauna as indicators.

Mancozeb is a dithiocarbamate non-systemic fungicide widely used to control fungal diseases of plants, commonly applied in apple orchards in Brazil. Instead of its common use, there are no reports about the risk to non-target organisms in Brazilian soils. We studied the risk of Mancozeb (in the commercial formulation Dithane ® NT) for standard invertebrate species (Folsomia candida, Eisenia andrei and Enchytraeus crypticus) in two subtropical Brazilian soils, Oxisol and Ultisol, which are representative of apple production areas in Brazil. Reproduction and survival tests were carried out following ISO guidelines. Results showed that Mancozeb in Oxisol reduced the survival and reproduction of collembolans (LC 50 54.43 and EC 50 2.72 mg a.i. kg -1 ) and enchytraeids (LC 50 6.97 and EC 50 3.56 mg a.i. kg -1 ), in lowest values than those observed in Ultisol (F. candida LC 50  > 1000 and EC 50  > 100 mg a.i. kg -1 ; E. crypticus LC 50 280.21 and EC 50 29.67). Effects to E. andrei were similar in both soils and indicated a lower sensitivity of this species to Mancozeb. The species F. candida and E. crypticus were more sensitive than E. andrei. These results reinforce the need to include other soil organisms besides earthworms, using chronical endpoints and considering different types of soils, to better predict the risk of pesticides for subtropical soils., (Copyright © 2019. Published by Elsevier Ltd.)

Inadequate application of liquid pig manure (LPM) may pose risks to the soil due to the potential contaminants that exists, as well as by the large water input that can originate excessive moisture. By using Terrestrial Model Ecosystems, this study aimed to evaluate the effect of application of LPM (82% moisture) using the application rates of 20, 50 and 150 m 3 ha -1 and also of the corresponding amount of water to understand the origin of effects on the soil fauna of two sub-tropical soils. In general, the results obtained for the two soil types indicated that LPM (150 m 3 ha -1 ) changed the composition of soil fauna, with an increase in the abundance of insect larvae and dipterans, but a decrease in the number of earthworms and enchytraeids. Microbial biomass, soil respiration and the nutrients Mg, K and P increased with the amount of LPM supplied to the soil. When analysing the effects of adding large volumes of water via the LPM, results showed that application did not originate significant effects on the parameters measured. The differences obtained when comparing both treatments were mainly attributed to the organic load brought by the LPM and not to the amount of water without LPM., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Arbuscular mycorrhizal fungi (AMF) are mutualistic symbionts considered a key group in soil systems involved in the provision of several ecosystem services. Recently they have been listed by EFSA as organisms to be included in the test battery for the risk assessment of plant protection product (PPPs). This study aimed to contribute to improve the ISO Protocol (ISO 10832: 2009) by assessing the feasibility of using other AMF species under different test conditions. Overall, results showed that AMF species Gigaspora albida and Rhizophagus clarus (selected out of five AMF species) are suitable to be used in spore germination tests using the ISO protocol (14 days incubation with sand or artificial soil as substrate) to test PPPs. However, several modifications to the protocol were made in order to accommodate the use of the tested isolates, namely the incubation temperature (28 °C instead of 24 °C) and the change of reference substance (boric acid instead of cadmium nitrate). The need for these changes, plus the results obtained with the three fungicides tested (chlorothalonil, mancozeb and metalaxyl-M) and comparisons made with literature on the relevance of the origin of AMF isolates in dictating the adequate test conditions, emphasize the importance of adjusting test conditions (AMF species/isolates and test temperature) when assessing effects for prospective risk assessment targeting different climatic zones. So, further studies should be conducted with different AMF species and isolates from different climatic regions, in order to better define which species/isolate and test conditions should be used to assess effects of a particular PPP targeting a given climatic zone.

Reference substances are recommended to evaluate the quality of laboratory test species and the reliability of ecotoxicity data. Boric acid (BA) has been recommended as reference substance in some standardized tests in OECD soil, but no data are available for Tropical Artificial Soil (TAS). For this purpose, avoidance tests with Eisenia andrei, lethality tests with E. andrei and Folsomia candida, and reproduction tests with E. andrei, Enchytraeus crypticus and F. candida were carried out in TAS (5% organic matter), following ISO guidelines, and compared between two laboratories. Collembolans were more sensitive than earthworms in lethality tests (LC 50  = 342 and > 1000 mg kg -1 , respectively). For both laboratories, the EC 50 values were similar for reproduction of oligochaeta species (165 mg kg -1 for E. crypticus; 242 and 281 mg kg -1 for E. andrei), but significantly different for reproduction of F. candida (96 and 198 mg kg -1 ). Present results suggest that boric acid could replace the current pesticides recommended by ISO guidelines as reference substances on reproduction tests with soil invertebrates in TAS. Concerning avoidance tests, additional investigations should be performed with other substances that cause no neurotoxic effects on soil organisms.

The effects of pig manure, from diets incorporating veterinary pharmaceuticals, on survival and reproduction of Folsomia candida were evaluated. Manures derived from the following diets: corn and soymeal (CS); 85% CS diet+15% wheat meal (TR); CS diet+100ppm doxycycline+50ppm colistin+2500ppm Zn oxide (CSa); TR diet+100ppm doxycycline+50ppm colistin+2500ppm Zn oxide (TRa). Manures were tested in two subtropical soils representative of southern (Oxisol and Entisol). Despite the antibiotics no significant differences were found between the four manures within each soil. However, strong differences were found on the toxicity between soils. In Oxisol, LC50 values were around 100m(3)ha(-1), and EC50 values around 80m(3)ha(-1). In Entisol these were much lower, with LC50 values oscillating around 20m(3)ha(-1) and EC50 values between 10-15m(3)ha(-1). The observed toxicity on both soils was attributed to excess of nitrogen, Cu and Zn in the highest doses. The strong difference between soils could be explained by soil properties, namely CEC, organic matter, and clay contents that were lower in Entisol, indicating a poor ability to retain contaminants increasing their availability in soil. Results suggest that the application of these residues should be regulated not only using a volume-based criterion, but should incorporate data on soil properties, complemented by an ecotoxicological assessment., (Copyright © 2016. Published by Elsevier B.V.)

This work aimed to evaluate density of associative diazotrophic bacteria populations in soil and grass root samples from heavy metal contaminated sites, and to characterize isolates from these populations, both, phenotypically (Zinc, Cadmium and NaCl tolerance in vitro, and protein profiles) and genotypically (16S rDNA sequencing), as compared to type strains of known diazotrophic species. Densities were evaluated by using NFb, Fam and JNFb media, commonly used for enrichment cultures of diazotrophic bacteria. Bacterial densities found in soil and grass root samples from contaminated sites were similar to those reported for agricultural soils. Azospirillum spp. isolates from contaminated sites and type strains from non-contaminated sites varied substantially in their in vitro tolerance to Zn+2 and Cd+2, being Cd+2 more toxic than Zn+2. Among the most tolerant isolates (UFLA 1S, 1R, S181, S34 and S22), some (1R, S34 and S22) were more tolerant to heavy metals than rhizobia from tropical and temperate soils. The majority of the isolates tolerant to heavy metals were also tolerant to salt stress as indicated by their ability to grow in solid medium supplemented with 30 g L(-1) NaCl. Five isolates exhibited high dissimilarity in protein profiles, and the 16S rDNA sequence analysis of two of them revealed new sequences for Azospirillum.