Assessing the reclamation of a contaminated site affected by the Fundão dam tailings trough phytoremediation and bioremediation.

The rupture of the Fundão dam (Brazil) spread tailings contaminated with sodium and ether-amine into the Doce River Basin. Aiming at rehabilitating a contaminated riparian site, phytoremediation with native species of the Atlantic Forest was performed under four treatments: ES-1: physical remediation (sediment scraping) + chemical remediation (organic matter) + bioremediation (double inoculation with the arbuscular mycorrhizal fungus (AMF) Rhizophagus irregularis and the plant growth-promoting bacteria Bacillus subtilis); ES-2: chemical remediation + bioremediation; ES-3: physical remediation + chemical remediation; ES-4: chemical remediation. Ether-amine and sodium contents, plant growth and, soil quality parameters were compared among treatments and relative to preserved and degraded sites. Two years after planting, the outstanding plant growth was attributed to the phytoremediation of ether-amine and ammonium, followed by a significant increase in soil microbial biomass (Phospholipid fatty acids-PLFAs), particularly the Gram+ bacteria and total fungi but not AMF, whose response was independent of the inoculation. While sodium and ether-amine declined, soil K, P, NO3- contents, dehydrogenase and acid phosphatase activities, cation exchange capacity (CEC) and soil aggregation increased, especially in ES-1. Thus, such remediation procedures are recommended for the restoration of riparian areas affected by the Fundão tailings, ultimately improving sediment fertility, aggregation and stabilization.

To the best of our knowledge, this is the only successful example of fragment of riparian Atlantic Forest successfully rehabilitated using native trees from the Atlantic Forest in the riparian zone affected by the contaminated Fundão dam tailings. For that, phytoremediation and bioremediation procedures were adopted to alleviate etheramine and sodium toxicity, resulting in the amelioration of soil fertility and particularly the stabilization of such highly disaggregated sediments, ultimately protecting the Doce river basin against these contaminants.

Rehabilitation of eroded trails and gullies on quartzite rock outcrops with native species in a high-altitude grassland.

The quartzite rock outcrops and the native vegetation of grasslands located at the Serra da Calçada Mountain in Minas Gerais State (Brazil) have been severely degraded by extreme sports activities such as motocross and off-road vehicles, greatly damaging the abundant headwaters. The main consequences thereof were hilly and gully erosion processes with soil loss and the deviation of the water from its original paths. However, currently, there is no report of successful restoration efforts in severely eroded outcrops in Brazilian high-altitude grasslands (campo rupestre). Through the Universal Soil Loss Equation (USLE), we found a high general erosion rate in the study site (669.91 t·ha-1·year-1), and the specific soil loss provoked by off-road vehicles on trails was significantly greater (49 m3 per 100 m2) than that caused by mountain bikes and trekking (5.8 m3 per 100 m2). We performed the physical reconstruction of eroded outcrops and surface water flow paths by allocating locally available quartzite rocks. These rocks were inoculated with different species of bryophytes and planted with native species under two treatments: un-inoculated and inoculated with arbuscular mycorrhizal fungi (AMF) spores of the Rhizophagus irregularis species. After 2 years, the bryophyte communities showed a similar pattern to the preserved site, and the AMF inoculation favoured plant establishment of most species, especially of the Asteraceae, Cyperaceae, Fabaceae, Malpighiaceae, Orchidaceae and Poaceae families. The AMF also improved the soil fertility, highlighting soil P, SOM, CEC, NH4+-N as well as soil water content and water retention capacity. Poaceae family species showed an outstanding occupation, which was considered a functional indicator of rehabilitation success, functioning as a "hydraulic carpet" for water exportation, conduction and drainage across the outcrops. This study provides an eco-technology to restore severely eroded outcrops over headwaters using native species in the Brazilian high-altitude grasslands.

Rehabilitation of a Riparian Site Contaminated by Tailings from the Fundão Dam, Brazil, Using Different Remediation Strategies.

The tailings spilled by the Fundão Dam rupture in the Doce River basin (Brazil) had a high pH, elevated sodium (Na) and ether amine, and low soil organic matter. With the aim of decreasing the toxic compounds, we established 2 remediation strategies: treatment 1, phytoremediation with tolerant native species of the Atlantic Forest cultivated on scraped sediment plus the incorporation of organic matter; and treatment 2, phytoremediation with native species plus superficial deposition of organic matter. The experimental site was compared with a degraded site that the dam tailings had reached and with a preserved site, a fragment of preserved Atlantic Forest. After 12 mo, plants showed an outstanding growth, especially after treatment 1 (~4 m), and the remediation procedures resulted in significant decreases in pH (from 8.0 to ~ 6.0), Na (from 154 to 22-35 mg/kg), electrical conductivity, and ether amine (from 6.0 to 0.5 mg/kg) in both treatments. By contrast, ammonium, a product of ether amine degradation, showed a significant increase in the experimental site, along with a significant increase in nitrate and improvement of soil microbial populations assessed by phospholipid fatty acid analysis. The treatments also improved soil fertility in the experimental site, as estimated by soil nutrients, cation exchange capacity, and soil aggregation. Based on the parameters analyzed, a principal component analysis showed that samples from the degraded site and the preserved site clustered in an opposite position and those from the experimental site clustered in an intermediate position but closer to the samples from the preserved site. Overall, our results demonstrated that the remediation procedures adopted were effective and resulted in rehabilitation of a riparian forest contaminated with dam tailings that included Na and ether amine. Environ Toxicol Chem 2021;40:2359-2373. © 2021 SETAC.

Synergistic effects between arbuscular mycorrhizal fungi and rhizobium isolated from As-contaminated soils on the As-phytoremediation capacity of the tropical woody legume Anadenanthera peregrina.

The synergistic relationship between arbuscular mycorrhizal fungi and plant species may play a key role in phytoremediation of arsenic(As)-contaminated soils. By using modified Leonard jars, we investigated both the distinct and integrative roles of arbuscular mycorrhizal fungi (AMF-Acaulospora scrobiculata) and rhizobia (BH-ICB-A8) isolated from As-contaminated soil on the capacity of Anadenanthera peregrina to reclaim arsenate [As(V)] from soil. AMF inoculation greatly increased plant phosphorous nutrition, as reflected in greater growth, and increased As-concentrations in the roots and shoots. While rhizobia inoculation alone increased nitrogen nutrition it did not promote plant growth or As-uptake. Rhizobia and AMF inoculation together had synergistic effects, however, increasing both the growth and the As-phytoremediation capacity of A. peregrina. Joint inoculation with rhizobia and AMF should therefore be considered a potential technique for rehabilitating As-contaminated areas using A. peregrina.

Remediation of a Riparian Site in the Brazilian Atlantic Forest Reached by Contaminated Tailings from the Collapsed Fundão Dam with Native Woody Species.

The failure of the Fundão dam in Brazil spilled alkaline contaminated sediments (pH ~8) into the Doce River basin. The contaminated sediments had high levels of ether amine (6 mg/kg) and Na (54 mg/kg) in relation to those in preserved sites. In the present study, a riparian forest was established over contaminated sediment using 2 main remediation strategies: phytoremediation with species native to the Atlantic forest and previously selected for their tolerance to sediment toxicity, and physicochemical remediation by incorporating organic matter (OM) into the sediment. In the experimental site (ES), the tolerant native woody species were cultivated in 2 treatments: T1, scraping of the sediments and incorporation of the OM, and T2, nonmanaged sediment and superficial deposition of the OM. The results were compared with the findings from a degraded site (DS), which was reached by the contaminated sediments but lacked vegetation, and a preserved site (PS), which was composed of a fragment of preserved Atlantic forest. Six months after transplanting, the plants from T1 showed a better height growth performance and survival index in relation to those from T2. Furthermore, T1 showed a significant decline in the pH (to ~6) concomitant with a reduction in the ether amine and Na contents (to ~0.4 mg/kg and 23 mg/kg, respectively). There was an improvement in the soil fertility and total microbial biomass in both treatments, especially in T1. Therefore, the adopted phyto- and physicochemical remediation procedures are recommended to reclaim zones reached by dam tailings containing Na and ether amine. Integr Environ Assess Manag 2020;16:669-675. © 2020 SETAC.

A ruptura da Barragen de Fundão em Mariana (Brazil) lançou um sedimento alcalino (pH ~8) ao longo da Bacia do Rio Doce. O sedimento contaminado apresentou altos níveis de éter-amina (6 mg/kg) e Na (54 mg/kg) em relação aqueles encontrados na área preservada. No presente estudo, uma floresta riparia foi estabelecida sobre este sedimento contaminado usando-se 2 estratégias de remediação: Fito-remediação com espécies nativas da Mata Atlântica, previamente selecionadas pela tolerância à toxicidade do sedimento, e remediação físico química pela raspagem superficial do sedimento e aplicação de matéria orgânica (OM). Na área experimental (ES) as espécies arbóreas da mata Atlântica selecionadas foram plantadas sob 2 tratamentos: T1, Raspagem superficial do sedimento e incorporação da OM, e T2, sedimento não foi manejado e recebeu aplicação superficial de OM. Os resultados foram comparados com aqueles obtidos na área degradada (DS), atingida pelo sedimento contaminado e sem vegetação, assim como com aqueles obtidos em uma área preservada (PS), constituída de um fragmento de Mata Atlântica. Após seis meses do plantio as plantas de T1 apresentaram um maior crescimento em altura e maior taxa de sobrevivência que as plantas de T2. Além disto ES, especialmente T1 mostraram um significante declínio do pH (~6.0) e concomitante redução nos teores de éter-amina e Na (~0.4 mg/kg e 23 mg/kg, respectivamente). Os procedimentos de remediação permitiram também um significativo aumento da fertilidade química e da biomassa microbiana do sedimento em ambos os tratamentos, especialmente em T1. Assim, os procedimentos de remediação físico-quimica e fitoremediação aqui adotados, são recomendados para recuperar as zonas atingidas pelo rejeito da barragem contaminado com sódio e éter-amina. Integr Environ Assess Manag 2020;16:669-675.

The N-fixing legume Periandra mediterranea constrains the invasion of an exotic grass (Melinis minutiflora P. Beauv) by altering soil N cycling.

Melinis minutiflora is an invasive species that threatens the biodiversity of the endemic vegetation of the campo rupestre biome in Brazil, displacing the native vegetation and favouring fire spread. As M. minutiflora invasion has been associated with a high nitrogen (N) demand, we assessed changes in N cycle under four treatments: two treatments with contrasting invasion levels (above and below 50%) and two un-invaded control treatments with native vegetation, in the presence or absence of the leguminous species Periandra mediterranea. This latter species was considered to be the main N source in this site due to its ability to fix N2 in association with Bradyrhizobia species. Soil proteolytic activity was high in treatments with P. mediterranea and in those severely invaded, but not in the first steps of invasion. While ammonium was the N-chemical species dominant in plots with native species, including P.mediterranea, soil nitrate prevailed only in fully invaded plots due to the stimulation of the nitrifying bacterial (AOB) and archaeal (AOA) populations carrying the amoA gene. However, in the presence of P. mediterranea, either in the beginning of the invasion or in uninvaded plots, we observed an inhibition of the nitrifying microbial populations and nitrate formation, suggesting that this is a biotic resistance strategy elicited by P. mediterranea to compete with M. minutiflora. Therefore, the inhibition of proteolytic activity and the nitrification process were the strategies elicited by P.mediterranea to constrain M.munitiflora invasion.

Invasion of the Brazilian campo rupestre by the exotic grass Melinis minutiflora is driven by the high soil N availability and changes in the N cycle.

The Serra do Rola Moça State Park (PESRM) in Minas Gerais State, Brazil is a preserved site representative of the campo rupestre biome over an ironstone outcrop that has a high level of plant diversity. Almost 60% of this grassy field has been invaded by the exotic molasses grass (Melinis minutiflora), which constitutes a severe threat to the biodiversity and survival of this biome, particularly due to the impacts of annual fires and inappropriate restoration interventions. Many invasive species exhibit a high demand for nitrogen (N). Hence, this work aimed to study the N cycle alterations promoted by M. minutiflora in a site of the campo rupestre, where the leguminous species Mimosa pogocephala was prevalent. The biome's soils exhibited a high natural N fertility and low C:N ratio. The main N source in this biome resulted from the biological N fixation performed by M. pogocephala associated with Burkholderia nodosa, as evidenced by the total leaf N content, leaf δ15N signature, nodule occupation and bacterial molecular identification analyses. The displacement of native species by molasses grass was associated with changes in the soil N forms, namely the nitrate increased as the ammonium decreased. The latter was the dominant N form in the native species plots, as observed in the soil analysis of total N, ammonium and nitrate contents. The dominant ammonium form was changed to the nitric form by the stimulation of ammonia-oxidising bacteria populations due to the invasive species. Therefore, the key mechanism behind the invasiveness of the exotic grass and the concomitant displacement of the native species may be associated with changes in the soil N chemical species. Based on this finding and on the high N-based soil fertility found in the campo rupestre N fertilisation procedures for restoration of invaded areas should be strictly avoided in this biome.

Nodulation in Dimorphandra wilsonii Rizz. (Caesalpinioideae), a threatened species native to the Brazilian Cerrado.

The threatened caesalpinioid legume Dimorphandra wilsonii, which is native to the Cerrado biome in Brazil, was examined for its nodulation and N(2)-fixing ability, and was compared with another, less-threatened species, D. jorgei. Nodulation and potential N(2) fixation was shown on seedlings that had been inoculated singly with five bradyrhizobial isolates from mature D. wilsonii nodules. The infection of D. wilsonii by two of these strains (Dw10.1, Dw12.5) was followed in detail using light and transmission electron microscopy, and was compared with that of D. jorgei by Bradyrhizobium strain SEMIA6099. The roots of D. wilsonii were infected via small transient root hairs at 42 d after inoculation (dai), and nodules were sufficiently mature at 63 dai to express nitrogenase protein. Similar infection and nodule developmental processes were observed in D. jorgei. The bacteroids in mature Dimorphandra nodules were enclosed in plant cell wall material containing a homogalacturonan (pectic) epitope that was recognized by the monoclonal antibody JIM5. Analysis of sequences of their rrs (16S rRNA) genes and their ITS regions showed that the five D. wilsonii strains, although related to SEMIA6099, may constitute five undescribed species of genus Bradyrhizobium, whilst their nodD and nifH gene sequences showed that they formed clearly separated branches from other rhizobial strains. This is the first study to describe in full the N(2)-fixing symbiotic interaction between defined rhizobial strains and legumes in the sub-family Caesalpinioideae. This information will hopefully assist in the conservation of the threatened species D. wilsonii.

Nodulation and nitrogen fixation by Mimosa spp. in the Cerrado and Caatinga biomes of Brazil.

*An extensive survey of nodulation in the legume genus Mimosa was undertaken in two major biomes in Brazil, the Cerrado and the Caatinga, in both of which there are high degrees of endemicity of the genus. *Nodules were collected from 67 of the 70 Mimosa spp. found. Thirteen of the species were newly reported as nodulating. Nodules were examined by light and electron microscopy, and all except for M. gatesiae had a structure typical of effective Mimosa nodules. The endosymbiotic bacteria in nodules from all of the Mimosa spp. were identified as Burkholderia via immunolabelling with an antibody against Burkholderia phymatum STM815. *Twenty of the 23 Mimosa nodules tested were shown to contain nitrogenase by immunolabelling with an antibody to the nitrogenase Fe- (nifH) protein, and using the delta(15)N ((15)N natural abundance) technique, contributions by biological N(2) fixation of up to 60% of total plant N were calculated for Caatinga Mimosa spp. *It is concluded that nodulation in Mimosa is a generic character, and that the preferred symbionts of Brazilian species are Burkholderia. This is the first study to demonstrate N(2) fixation by beta-rhizobial symbioses in the field.

Seleçäo e caracterizaçäo de estirpes de rhizobium estáveis e capazes de fixar nitrogênio em feijäo(phaseolus vulgaris L.) / Selection and characterization of rhizobium spp.strains stable and capable in fixing nitrogen in bean(phaseolus vulgaris L.)

resumo:Determinou-se a variabilidade na capacidade de fixaçäo de N2, através de testes de reduçäo de acetileno em nódulos formados por estirpes de Rhizobium, antes e após exposiçäo das bactérias "in vitro" à temperaturas elevadas (38-39ºC).Nódulos formados tanto por estirpes de R.leguminosarum bv.phaseoli como R.tropici mais tolerantes a altas temperaturas quando inoculadas em feijäo, näo sofreram alteraçöes nas características simbióticas tais como, atividade de nitrogenase, peso seco de planta e nitrogênio total fixado.O padräo de proteínas(eletroforese SDS-PAGE) diferenciou estirpes entre e dentro das espécies.A hibridizaçäo do DNA total usando "nifprob" marcada via "nick translation"(biotina 14 dATP), quando a digestäo foi efetuada com EcoRI, diferenciou a espécie de R.leguminosarumbv.phaseoli de R.tropici.Foi observado polimorfismo entre as estirpes de R.leguminosarum bv.phaseoli após digestäo com Bam HI e entre R.leguminosarum bv.phaseoli e R.tropici após a digestäo com Hind III.Näo foram detectadas alteraçöes nos padröes protéicos ou genômicos e na atividade da nitrogenase da mesma estirpe antes e após crescimento a temperaturas elevadas, indicando que as estirpes de ambas as espécies(R.leguminosarum bv.phaseoli e R.tropici), tolerantes a altas temperaturas säo também mais estáveis geneticamente