Exposure to fipronil, 2,4-D and vinasse influences macroinvertebrate assemblage structure: An experimental mesocosm approach.

Sugarcane is one of the main monocultures in Brazil and widely uses herbicide 2,4-D and fipronil insecticide. In addition, vinasse can be mentioned as it is widely used in this plantation. These compounds occurring simultaneously in the aquatic environment can potentiate the deleterious effects on organisms. Thus, this study aimed to evaluate the composition, abundance and ecological indices of the benthic macroinvertebrate community, as well as its ability to reestablish itself considering environmental contamination by the pesticides Regent® 800WG (active ingredient - a.i. fipronil) (F) and DMA® 806BR (a.i. 2,4-D) (D) and vinasse (V), alone and in mixtures: pesticides - M and the three contaminants - MV. The study was conducted using open-air mesocosms. The macroinvertebrate community was monitored by colonization structures, the physical-chemical parameters, metals and pesticides were determined and the effects of contaminants were evaluated over the exposure time in 1, 7, 14, 28, 75 to 150 days. A multiple regression was performed between the water parameters and significant relationships were found between parameters associated with vinasse contamination (pH, total nitrogen, turbidity, and dissolved oxygen) and concentration of fipronil and the ecological variables studied. Over time, changes were observed in the composition of the community. The dominance and richness increased in treatments V and MV. The family Chironomidae and subclass Oligochaeta were more sensitive to the treatment V and MV, while individuals from the families Phoridae, Ephydridae and Sciomyzidae were occasionally found (depending on the experimental time) in these treatments. The insects were sensitive to treatments F and M, disappearing in these mesocosms after contamination, reappearing only after 75 days. The results reveal that sugarcane management practices associated with the use of pesticides and vinasse as fertilizer pose risks to the macroinvertebrate community with consequences for the trophic chains, given its importance in freshwater ecosystems and adjacent terrestrial environments.

Detection and estimation of Mastigimas anjosi (Hemiptera: Calophyidae) populations on Cedrela fissilis trees.

Mastigimas anjosi is an important pest of Cedrela fissilis in the southeastern and southern regions of Brazil. The objective of the present study was to investigate the effect of the temperature, relative humidity, rainfall and natural enemies on the flight activity of adults and the movement of M. anjosi nymphs, with two sampling methods, yellow sticky traps and direct collections on the leaves of C. fissilis trees. The sex ratio of this pest was also assessed. The number of M. anjosi individuals was negatively correlated with the minimum and maximum temperatures, with a population peak in late May and early June 2017. The numbers of individuals of this psyllid was positively correlated with the relative humidity and rainfall. Larvae and adults of Cycloneda sanguinea, Hippodamia convergens adults and Atopozelus opsimus nymphs and adults preyed upon M. anjosi adults and nymphs on C. fissilis leaves in the field. Mastigimas anjosi sex ratio was 0.46. Information about sampling methods of M. anjosi and its natural enemies is presented and can contribute to the integrated management of this pest in the field.

Methane emission suppression in flooded soil from Amazonia.

The coupling between ferrous iron and methane production has important global implications, with iron ions acting as electron acceptors for anaerobic oxidation of methane (AOM) and inhibitors of methanogenesis in different environments, including floodplain soils. In this sense, we analyzed the relationship between Fe(II) concentration and methane production in soil layers collected at 0-15 cm and 15-30 cm from flooded-forest and -agroforestry in Amazonian clear water floodplain incubated in anaerobic batch reactors using acetate, formate and glucose as organic sources. High throughput sequencing of archaeal and bacterial 16S rRNA genes was employed to assess the abundance and composition of the active methanogenic and methanotrophic microbial groups potentially involved in Fe(III)-dependent AOM in the soil used as inoculum. Positive correlation was revealed between Fe(II) concentration and methane production, with higher accumulation of Fe(II) in incubated soil layer collected at 0-15 cm in both forest and agroforestry sites for all the three organic sources. The accumulation of Fe(II) in the incubated soil evidenced the oxidation of Fe(III) potentially by Methanobacterium, Desulfobulbus and 'Candidatus methanoperedens nitroreducens' living in anaerobic condition at this soil layer. The results point out to the microbial ferric iron reduction as an important potential pathway for anaerobic organic matter decomposition in Amazonian floodplain, evidencing methanogenesis suppression by Fe(III) reduction in flooded-forest and -agroforestry in Amazonian clear water river floodplain.

Bacterial community composition and diversity of two different forms of an organic residue of bioenergy crop.

The use of residue of sugarcane ethanol industry named vinasse in fertirrigation is an established and widespread practice in Brazil. Both non-concentrated vinasse (NCV) and concentrated vinasse (CV) are used in fertirrigation, particularly to replace the potassium fertilizer. Although studies on the chemical and organic composition of vinasse and their impact on nitrous oxide emissions when applied in soil have been carried out, no studies have evaluated the microbial community composition and diversity in different forms of vinasse. We assessed the bacterial community composition of NCV and CV by non-culturable and culturable approaches. The non-culturable bacterial community was assessed by next generation sequencing of the 16S rRNA gene and culturable community by isolation of bacterial strains and molecular and biochemical characterization. Additionally, we assessed in the bacterial strains the presence of genes of nitrogen cycle nitrification and denitrification pathways. The microbial community based on 16S rRNA sequences of NCV was overrepresented by Bacilli and Negativicutes while CV was mainly represented by Bacilli class. The isolated strains from the two types of vinasse belong to class Bacilli, similar to Lysinibacillus, encode for nirK gene related to denitrification pathway. This study highlights the bacterial microbial composition particularly in CV what residue is currently recycled and recommended as a sustainable practice in sugarcane cultivation in the tropics.

Correction to: Genome-resolved metagenomics of sugarcane vinasse bacteria.

[This corrects the article DOI: 10.1186/s13068-018-1036-9.].

Methodology for soil respirometric assays: Step by step and guidelines to measure fluxes of trace gases using microcosms.

This methodology is proposed to measure the fluxes of trace gases among microcosms and the atmosphere. As microcosm respiration we include both aerobic and anaerobic respiration, which may results in CO2, CH4, NO, N2O, N2, H2S and H2 fluxes. Its applicability includes the assessment of products biodegradability and toxicity, the effect of treatments and products on greenhouse gases fluxes, and the mineralization of organic fertilizers. A step by step procedure; the complementary parameters and good practices that might be taken into account to perform a microcosm experiment; and the tools nowadays available that could be useful in this respirometric methodology are presented. We included a spreadsheet with calculus examples. Samples were taken at 1; 30; 60 and 90 min after closing the microcosms to determine the gases fluxes. The dilution effect was negligible, as we present. Besides CO2, we have successfully quantified the fluxes of CH4 and N2O from the microcosms in a broad range of concentrations. This method is useful in technical and scientific studies, for instances to test new products and improve the understanding of microbial processes, respectively. •Simple materials are required to set up the microcosm.•Examples of (pre) treatments are given regarding water availability, fertilizer doses, pH adjustment and nutrients amendments.•The method was suitable to directly measure multiple trace gases fluxes, either produced or consumed during microcosm respiration.

Genome-resolved metagenomics of sugarcane vinasse bacteria.

BACKGROUND: The production of 1 L of ethanol from sugarcane generates up to 12 L of vinasse, which is a liquid waste containing an as-yet uncharacterized microbial assemblage. Most vinasse is destined for use as a fertilizer on the sugarcane fields because of the high organic and K content; however, increased N2O emissions have been observed when vinasse is co-applied with inorganic N fertilizers. Here we aimed to characterize the microbial assemblage of vinasse to determine the gene potential of vinasse microbes for contributing to negative environmental effects during fertirrigation and/or to the obstruction of bioethanol fermentation. RESULTS: We measured chemical characteristics and extracted total DNA from six vinasse batches taken over 1.5 years from a bioethanol and sugar mill in Sao Paulo State. The vinasse microbial assemblage was characterized by low alpha diversity with 5-15 species across the six vinasses. The core genus was Lactobacillus. The top six represented bacterial genera across the samples were Lactobacillus, Megasphaera and Mitsuokella (Phylum Firmicutes, 35-97% of sample reads); Arcobacter and Alcaligenes (Phylum Proteobacteria, 0-40%); Dysgonomonas (Phylum Bacteroidetes, 0-53%); and Bifidobacterium (Phylum Actinobacteria, 0-18%). Potential genes for denitrification but not nitrification were identified in the vinasse metagenomes, with putative nirK and nosZ genes the most represented. Binning resulted in 38 large bins with between 36.0 and 99.3% completeness, and five small mobile element bins. Of the large bins, 53% could be classified at the phylum level as Firmicutes, 15% as Proteobacteria, 13% as unknown phyla, 13% as Bacteroidetes and 6% as Actinobacteria. The large bins spanned a range of potential denitrifiers; moreover, the genetic repertoires of all the large bins included the presence of genes involved in acetate, CO2, ethanol, H2O2, and lactose metabolism; for many of the large bins, genes related to the metabolism of mannitol, xylose, butyric acid, cellulose, sucrose, "3-hydroxy" fatty acids and antibiotic resistance were present based on the annotations. In total, 21 vinasse bacterial draft genomes were submitted to the genome repository. CONCLUSIONS: Identification of the gene repertoires of vinasse bacteria and assemblages supported the idea that organic carbon and nitrogen present in vinasse together with microbiological variation of vinasse might lead to varying patterns of N2O emissions during fertirrigation. Furthermore, we uncovered draft genomes of novel strains of known bioethanol contaminants, as well as draft genomes unknown at the phylum level. This study will aid efforts to improve bioethanol production efficiency and sugarcane agriculture sustainability.

Nitrous oxide emission related to ammonia-oxidizing bacteria and mitigation options from N fertilization in a tropical soil.

Nitrous oxide (N2O) from nitrogen fertilizers applied to sugarcane has high environmental impact on ethanol production. This study aimed to determine the main microbial processes responsible for the N2O emissions from soil fertilized with different N sources, to identify options to mitigate N2O emissions, and to determine the impacts of the N sources on the soil microbiome. In a field experiment, nitrogen was applied as calcium nitrate, urea, urea with dicyandiamide or 3,4 dimethylpyrazone phosphate nitrification inhibitors (NIs), and urea coated with polymer and sulfur (PSCU). Urea caused the highest N2O emissions (1.7% of N applied) and PSCU did not reduce cumulative N2O emissions compared to urea. NIs reduced N2O emissions (95%) compared to urea and had emissions comparable to those of the control (no N). Similarly, calcium nitrate resulted in very low N2O emissions. Interestingly, N2O emissions were significantly correlated only with bacterial amoA, but not with denitrification gene (nirK, nirS, nosZ) abundances, suggesting that ammonia-oxidizing bacteria, via the nitrification pathway, were the main contributors to N2O emissions. Moreover, the treatments had little effect on microbial composition or diversity. We suggest nitrate-based fertilizers or the addition of NIs in NH4(+)-N based fertilizers as viable options for reducing N2O emissions in tropical soils and lessening the environmental impact of biofuel produced from sugarcane.

Does Diatomaceous Earth Control Leaf-Cutter Ants (Hymenoptera: Formicidae) in the Eucalyptus Plantations?

Genus Atta includes some of the most important Formicidae leaf cutter ants which cause extensive damage to the eucalyptus plantations. Atta sexdens rubropilosa Forel, one of the chief pests in Brazilian reforestation, can restrict and reduce forest productivity by its intense and constant leaf-cutting activities on plants at all stages. Therefore, the demand for new products to control A. sexdens rubropilosa indicates the study of the utilization of the dry powder formulation of diatomaceous earth (DE) against this pest in the eucalyptus cultivars. The study was conducted using 120 colonies of A. sexdens rubropilosa in Eucalyptus grandis Hill ex. Maiden x Eucalyptus urophylla Blake (Myrtaceae) (urograndis) stand. The randomized block experimental design was used with six treatments (1, 10, 25, and 50 g/m2 of DE, 6.0 g/m2 sulfluramid bait per square meter of loose soil, and the control) with five replications, each with four colonies of this ant. Diatomaceous earth was applied to the active A. sexdens rubropilosa ant holes, and the sulfluramid bait was applied in bulk in a localized manner. The control efficacy of A. sexdens rubropilosa with DE was low, showing values similar to that of the control, and, for this reason, it cannot be used to control this ant. The bait with sulfluramid showed higher efficacy than those of the other treatments.

Enhanced-efficiency fertilizers in nitrous oxide emissions from urea applied to sugarcane.

The environmental benefits of producing biofuels from sugarcane have been questioned due to greenhouse gas emissions during the biomass production stage, especially nitrous oxide (NO) associated with nitrogen (N) fertilization. The objective of this work was to evaluate the use of nitrification inhibitors (NIs) dicyandiamide (DCD) and 3,4 dimethylpyrazole phosphate (DMPP) and a controlled-release fertilizer (CRF) to reduce NO emissions from urea, applied at a rate of 120 kg ha of N. Two field experiments in ratoon cycle sugarcane were performed in Brazil. The treatments were (i) no N (control), (ii) urea, (iii) urea+DCD, (iv) urea+DMPP, and (v) CRF. Measurements of NO fluxes were performed using static chambers with four replications. The measurements were conducted three times per week during the first 3 mo and biweekly afterward for a total of 217 and 382 d in the first and second seasons, respectively. The cumulative NO-N emissions in the first ratoon cycle were 1098 g ha in the control treatment and 1924 g ha with urea (0.7% of the total N applied). Addition of NIs to urea reduced NO emissions by more than 90%, which did not differ from those of the plots without N. The CRF treatment showed NO emissions no different from those of urea. The results were similar in the second ratoon: the treatment with urea showed NO emissions of 0.75% of N applied N. Application of NIs resulted in a strong reduction in NO emissions, but CRF increased emissions compared with urea. We therefore conclude that both NIs can be options for mitigation of greenhouse gas emission in sugarcane used for bioenergy.

Stocks of carbon and nitrogen and partitioning between above- and belowground pools in the Brazilian coastal Atlantic Forest elevation range.

We estimated carbon and nitrogen stocks in aboveground biomass (AGB) and belowground biomass (BGB) along an elevation range in forest sites located on the steep slopes of the Serra do Mar on the north coast of the State of São Paulo, southeast Brazil. In elevations of 100 m (lowland), 400 m (submontane), and 1000 m (montane) four 1-ha plots were established, and above- (live and dead) and belowground (live and dead) biomass were determined. Carbon and nitrogen concentrations in each compartment were determined and used to convert biomass into carbon and nitrogen stocks. The carbon aboveground stock (C(AGB)) varied along the elevation range from approximately 110 to 150 Mg·ha(-1), and nitrogen aboveground stock (N(AGB)), varied from approximately 1.0 to 1.9 Mg·ha(-1). The carbon belowground stock (C(BGB)) and the nitrogen belowground stock (N(BGB)) were significantly higher than the AGB and varied along the elevation range from approximately 200-300 Mg·ha(-1), and from 14 to 20 Mg·ha(-1), respectively. Finally, the total carbon stock (C(TOTAL)) varied from approximately 320 to 460 Mg·ha(-1), and the nitrogen total stock (N(TOTAL)) from approximately 15 to 22 Mg·ha(-1). Most of the carbon and nitrogen stocks were found belowground and not aboveground as normally found in lowland tropical forests. The above- and belowground stocks, and consequently, the total stocks of carbon and nitrogen increased significantly with elevation. As the soil and air temperature also decreased significantly with elevation, we found a significantly inverse relationship between carbon and nitrogen stocks and temperature. Using this inverse relationship, we made a first approach estimate that an increase of 1°C in soil temperature would decrease the carbon and nitrogen stocks in approximately 17 Mg·ha(-1) and 1 Mg·ha(-1) of carbon and nitrogen, respectively.