RESUMO
Lindane is a toxic and persistent organochlorine pesticide, whose extensive use generated its accumulation in different environmental matrices. Bioremediation is a promising technology that can be used combining bioaugmentation and biostimulation processes to soil restoration. The aim of the present work was to determine the conditions of maximum lindane removal by bioaugmentation with an actinobacteria consortium and biostimulation with sugarcane filter cake (SCFC). The assays were carried out on lindane-contaminated silty loam (SLS), clayey (CS), and sandy (SS) soils. Through complete factorial designs, the effects of three abiotic factors (moisture content, proportion and size of SCFC particles) were evaluated on lindane removal. In addition, a response optimizer determined the optimal conditions for pesticide removal in bioaugmented and biostimulated soils, in the range of levels studied for each factor. In these conditions, bioaugmentation of biostimulated soils increased the pesticide removal (SLS: 61.4%, CS: 70.8%, SS: 86.3%), heterotrophic microbial counts, and soil enzymatic activities, and decreased lindane T1/2, regarding the non-bioaugmented biostimulated controls, after 14 days of assay. The values of these parameters confirmed the efficiency of the bioremediation process. Finally, the viability of the four strains was demonstrated at the end of the assay. The results indicate that the simultaneous application of bioaugmentation with the actinobacteria consortium and biostimulation with SCFC constitutes a promising tool for restoring soils contaminated with lindane, by using the optimal conditions obtained through the factorial designs.
Assuntos
Biodegradação Ambiental , Hexaclorocicloexano/metabolismo , Microbiologia do Solo , Poluentes do Solo/metabolismo , Actinobacteria , Bactérias , Hexaclorocicloexano/análise , Praguicidas , Saccharum , Solo , Poluentes do Solo/análiseRESUMO
Lindane is an organochlorine pesticide that, due to its persistence in the environment, is still detected in different matrices. Bioremediation using actinobacteria consortia proved to be promising for the restoration of contaminated soils. Another alternative to remove xenobiotics is to use agricultural residues, which stimulates microbial activity, increasing its capacity to degrade organic pollutants. The present work studies the coupling of sugarcane bagasse biostimulation and bioaugmentation with the actinobacteria consortium composed of Streptomyces sp. A2, A5, A11 and M7 on lindane removal in different soil types. In this sense, factorial designs with three factors (proportion and size of sugarcane bagasse particles, and moisture content) were employed. A response optimizer identified the combination of factors levels that jointly allowed obtaining the maximum lindane removal in the evaluated conditions. In the optimal conditions, the effect of the bioremediation process on soil microbiota was studied by evaluating different parameters. The highest lindane removal percentages were detected in biostimulated microcosms bioaugmented with the microbial consortium, which were accompanied by a decrease in lindane half-life respect to the controls. Also, the bioaugmentation of biostimulated microcosms increased the microbial counts and enhanced soil enzymatic activities, corroborating the bioremediation process efficiency. The survival of the four actinobacteria at the end of the assay confirmed the ability of all Streptomyces strains to colonize amended soils. Bioremediation by simultaneous application of biostimulation with sugarcane bagasse and bioaugmentation with the actinobacteria consortium, in the optimized conditions, represents an efficient strategy to restore lindane contaminated soils.
Assuntos
Hexaclorocicloexano/isolamento & purificação , Hexaclorocicloexano/metabolismo , Poluentes do Solo/isolamento & purificação , Poluentes do Solo/metabolismo , Solo/química , Streptomyces/efeitos dos fármacos , Streptomyces/metabolismo , Biodegradação Ambiental/efeitos dos fármacos , Celulose/química , Celulose/farmacologia , Consórcios Microbianos/efeitos dos fármacos , Saccharum/químicaRESUMO
Bioremediation using actinobacterium consortia proved to be a promising alternative for the purification of co-contaminated environments. In this sense, the quadruple consortium composed of Streptomyces sp. M7, MC1, A5, and Amycolatopsis tucumanensis AB0 has been able to remove significant levels of Cr(VI) and lindane from anthropogenically contaminated soils. However, the effectiveness of the bioremediation process could not be evaluated only by analytical monitoring, which is complex mainly due to the characteristics of the matrix, producing non-quantitative analyte recoveries, or interferences in the detection stage and quantification. However, the effectiveness of the bioremediation process cannot be evaluated only through analytical monitoring, which is complex due mainly to the characteristics of the matrix, to the recoveries of non-quantitative analytes or to interferences in the detection and quantification stage. For this reason, it is essential to have tools of ecological relevance to assess the biological impact of pollutants on the environment. In this context, the objective of this work was to establish the appropriate bioassays to evaluate the effectiveness of a bioremediation process of co-contaminated soils. For this, five model species were studied: four plant species (Lactuca sativa, Raphanus sativus, Lycopersicon esculentum, and Zea mays) and one animal species (Eisenia fetida). On plant species, the biomarkers evaluated were inhibition of germination (IG) and the length of hypocotyls/steam and radicles/roots of the seedling. While on E. fetida, mortality (M), weight lost, coelomocyte concentration and cell viability were tested. These bioindicators and the battery of biomarkers quantified in them showed a different level of sensitivity, from maximum to minimum: E. fetidaâ¯>â¯L. esculentumâ¯>â¯L. sativaâ¯>â¯R. sativus â«>Z. mays. Therefore, E. fetida and L. esculentum and their respective biomarkers were selected to evaluate the effectiveness of the bioremediation process due to the capability of assessing the effect on the flora and the fauna of the soil, respectively. The joint application of these bioindicators in a field scale bioremediation process is a feasible tool to demonstrate the recovery of the quality and health of the soil.
Assuntos
Actinobacteria/metabolismo , Cromo/toxicidade , Hexaclorocicloexano/toxicidade , Poluentes do Solo/toxicidade , Animais , Biodegradação Ambiental , Cromo/metabolismo , Biomarcadores Ambientais , Hexaclorocicloexano/metabolismo , Consórcios Microbianos , Oligoquetos/efeitos dos fármacos , Desenvolvimento Vegetal/efeitos dos fármacos , Espécies Sentinelas , Poluentes do Solo/metabolismo , Streptomyces/metabolismoRESUMO
Highly contaminated γ-hexachlorocyclohexane (lindane) areas were reported worldwide. Low aqueous solubility and high hydrophobicity make lindane particularly resistant to microbial degradation. Physiological and genetic Streptomyces features make this genus more appropriate for bioremediation compared with others. Complete degradation of lindane was only proposed in the genus Sphingobium although the metabolic context of the degradation was not considered. Streptomyces sp.M7 has demonstrated ability to remove lindane from culture media and soils. In this study, we used MS-based label-free quantitative proteomic, RT-qPCR and exhaustive bioinformatic analysis to understand lindane degradation and its metabolic context in Streptomyces sp. M7. We identified the proteins involved in the up-stream degradation pathway. In addition, results demonstrated that mineralization of lindane is feasible since proteins from an unusual down-stream degradation pathway were also identified. Degradative steps were supported by an active catabolism that supplied energy and reducing equivalents in the form of NADPH. To our knowledge, this is the first study in which degradation steps of an organochlorine compound and metabolic context are elucidate in a biotechnological genus as Streptomyces. These results serve as basement to study other degradative actinobacteria and to improve the degradation processes of Streptomyces sp. M7.
Assuntos
Hexaclorocicloexano/metabolismo , Redes e Vias Metabólicas , Proteoma/metabolismo , Proteômica/métodos , Streptomyces/genética , Streptomyces/metabolismo , Transcriptoma , Biodegradação Ambiental , Proteoma/análiseRESUMO
Actinobacteria are well-known degraders of toxic materials that have the ability to tolerate and remove organochloride pesticides; thus, they are used for bioremediation. The biodegradation of organochlorines by actinobacteria has been demonstrated in pure and mixed cultures with the concomitant production of metabolic intermediates including γ-pentachlorocyclohexene (γ-PCCH); 1,3,4,6-tetrachloro-1,4-cyclohexadiene (1,4-TCDN); 1,2-dichlorobenzene (1,2-DCB), 1,3-dichlorobenzene (1,3-DCB), or 1,4-dichlorobenzene (1,4-DCB); 1,2,3-trichlorobenzene (1,2,3-TCB), 1,2,4-trichlorobenzene (1,2,4-TCB), or 1,3,5-trichlorobenzene (1,3,5-TCB); 1,3-DCB; and 1,2-DCB. Chromatography coupled to mass spectrometric detection, especially GC-MS, is typically used to determine HCH-isomer metabolites. The important enzymes involved in HCH isomer degradation metabolic pathways include hexachlorocyclohexane dehydrochlorinase (LinA), haloalkane dehalogenase (LinB), and alcohol dehydrogenase (LinC). The metabolic versatility of these enzymes is known. Advances have been made in the identification of actinobacterial haloalkane dehydrogenase, which is encoded by linB. This knowledge will permit future improvements in biodegradation processes using Actinobacteria. The enzymatic and genetic characterizations of the molecular mechanisms involved in these processes have not been fully elucidated, necessitating further studies. New advances in this area suggest promising results. The scope of this paper encompasses the following: (i) the aerobic degradation pathways of hexachlorocyclohexane (HCH) isomers; (ii) the important genes and enzymes involved in the metabolic pathways of HCH isomer degradation; and (iii) the identification and quantification of intermediate metabolites through gas chromatography coupled to mass spectrometry (GC-MS).
Assuntos
Biodegradação Ambiental , Hexaclorocicloexano , Streptomyces , Biotecnologia , Hexaclorocicloexano/química , Hexaclorocicloexano/isolamento & purificação , Hexaclorocicloexano/metabolismo , Isomerismo , Streptomyces/química , Streptomyces/metabolismo , Streptomyces/fisiologiaRESUMO
Environmental mixed pollution by both organic and inorganic compounds are detected worldwide. Phytoremediation techniques have been proposed as ecofriendly methods for cleaning up polluted sites. Several studies have demonstrated enhanced dissipation of contaminants at the root-soil interface through an increase in microbial activity caused by the release of plant root exudates (REs). The aim of this study was to evaluate the effectiveness for Cr(VI) and lindane removal by Streptomyces M7 cultured in a co-contaminated system in presence of maize REs. Our results showed when REs were added to the contaminated minimal medium (MM) as the only carbon source, microbial removal of Cr(VI) and lindane increased significantly in comparison to contaminant removal obtained in MM with glucose 1 g L-1 . The maximum removal of 91% of lindane and 49.5% of Cr(VI) were obtained in the co-contaminated system. Moreover, Streptomyces M7 showed plant growth promoting traits which could improve plant performance in contaminated soils. The results presented in this study provide evidence that maize REs improved growth of Streptomyces M7 when REs were used as a carbon source in comparison to glucose. Consequently, lindane and Cr(VI) removal was considerably enhanced making evident the phytoremediation potential of the actinobacteria-plant partnership.
Assuntos
Carcinógenos Ambientais/metabolismo , Cromo/metabolismo , Hexaclorocicloexano/metabolismo , Inseticidas/metabolismo , Exsudatos de Plantas/metabolismo , Streptomyces/metabolismo , Zea mays/metabolismo , Carbono/metabolismo , Meios de Cultura/química , Exsudatos de Plantas/isolamento & purificação , Raízes de Plantas/metabolismo , Streptomyces/crescimento & desenvolvimentoRESUMO
Concentrations of organochlorine pesticides were quantified in samples of feathers (n = 17) and blood (n = 15) of the ferruginous pygmy owl (Glaucidium brasilianum). The individuals were captured near the Protected Natural Area Cerro Sonsonate, Chiapas, Mexico, between February and June 2014. In both tissues, pesticides belonging to seven organochlorine chemical families were detected. However, the organochlorine pesticide concentrations differed between feathers and blood. The highest concentrations of hexachlorocyclohexanes were found in feathers (0.63 ± 0.89 µg/g), whereas the highest concentrations of ΣDrines were found in blood (0.31 ± 0.47 µg/mL). By using the summed concentrations for each of the seven families of pesticides found in feathers, we did not find any significant correlation between the pesticides and pectoral muscle or body weight (p > 0.15). The ΣDDT group was the only pesticide family that showed a positive correlation with owl body weight (r = 0.60, p = 0.05); the concentrations of these pesticides were also high in feather and blood tissues (r = 0.87, p = 0.02). Our results confirm that ferruginous pygmy owls in the study area are exposed to these pesticides.
Assuntos
Monitoramento Ambiental , Poluentes Ambientais/metabolismo , Praguicidas/metabolismo , Estrigiformes/metabolismo , Animais , Plumas/química , Plumas/metabolismo , Hexaclorocicloexano/sangue , Hexaclorocicloexano/metabolismo , Hidrocarbonetos Clorados/análise , Hidrocarbonetos Clorados/metabolismo , México , Praguicidas/análiseRESUMO
The organochlorine insecticide γ-hexachlorocyclohexane (γ-HCH, lindane) and its non-insecticidal α- and ß-isomers continue to pose serious environmental and health concerns, although their use has been restricted or completely banned for decades. In this study we report the first evidence of the growth ability of a Streptomyces strain in a mineral salt medium containing high doses of α- and ß-HCH (16.6 mg l(-1)) as a carbon source. Degradation of HCH isomers by Streptomyces sp. M7 was investigated after 1, 4, and 7 days of incubation, determining chloride ion release, and residues in the supernatants by GC with µECD detection. The results show that both the α- and ß-HCH isomers were effectively metabolized by Streptomyces sp. M7, with 80 and 78 % degradation respectively, after 7 days of incubation. Moreover, pentachlorocyclohexenes and tetrachlorocyclohexenes were detected as metabolites. In addition, the formation of possible persistent compounds such as chlorobenzenes and chlorophenols were studied by GC-MS, while no phenolic compounds were detected. In conclusion, we have demonstrated for the first time that Streptomyces sp. M7 can degrade α- and ß-isomers individually or combined with γ-HCH and could be considered as a potential agent for bioremediation of environments contaminated by organochlorine isomers.
Assuntos
Hexaclorocicloexano/química , Hexaclorocicloexano/metabolismo , Poluentes do Solo/química , Poluentes do Solo/metabolismo , Streptomyces/metabolismo , Anaerobiose , Biodegradação Ambiental , Inseticidas/química , Inseticidas/metabolismo , IsomerismoRESUMO
Banana has been a main agricultural product in the French West Indies (Guadeloupe and Martinique) since the 1960s. This crop requires the intensive use of pesticides to prevent attacks by insect pests. Chlorinated pesticides, such as hexachlorocyclohexane (HCH), chlordecone and dieldrin, were used until the beginning of the 1990s, resulting in a generalized diffuse contamination of the soil and water in the areas of banana production, hence the need to develop solutions for cleanup of the polluted sites. The aims of this work were (i) to assess lindane degradation in soil slurry microcosms treated with lindane at 10 mg/L and (ii) to detect the catabolic genes involved in the HCH degradation pathway. The soil slurry microcosm system showed a 40% lindane degradation efficiency at the end of a 30-day experiment. Lower lindane removal was also detected in the abiotic controls, probably caused by pesticide adsorption to soil particles. Indeed, the lindane concentration decreased from 6000 to 1330 ng/mL and from 800 to 340 ng/mL for the biotic and abiotic soils, respectively. Nevertheless, some of the genes involved in the HCH degradation pathway were amplified by polymerase chain reaction (PCR) from crude deoxyribonucleic acid (DNA) extracted from the Guadeloupe agricultural soil, suggesting that HCH degradation is probably mediated by bacteria closely related to the family Sphingomonadaceae.
Assuntos
Hexaclorocicloexano/metabolismo , Inseticidas/metabolismo , Microbiologia do Solo , Agricultura , Biodegradação Ambiental , Guadalupe , Redes e Vias Metabólicas , Poluentes do Solo/metabolismo , Sphingomonadaceae/metabolismoRESUMO
The aim of this work was to study the impact of environmental factors on the bioremediation of Cr(VI) and lindane contaminated soil, by an actinobacterium, Streptomyces sp. M7, in order to optimize the process. Soil samples were contaminated with 25 µg kg(-1) of lindane and 50 mg kg(-1) of Cr(VI) and inoculated with Streptomyces sp. M7. The lowest inoculum concentration which simultaneously produced highest removal of Cr(VI) and lindane was 1 g kg(-1). The influence of physical and chemical parameters was assessed using a full factorial design. The factors and levels tested were: Temperature: 25, 30, 35°C; Humidity: 10%, 20%, 30%; Initial Cr(VI) concentration: 20, 50, 80 mg kg(-1); Initial lindane concentration: 10, 25, 40 µg kg(-1). Streptomyces sp. M7 exhibited strong versatility, showing the ability to bioremediate co-contaminated soil samples at several physicochemical conditions. Streptomyces sp. M7 inoculum size was optimized. Also, it was fitted a model to study this process, and it was possible to predict the system performance, knowing the initial conditions. Moreover, optimum temperature and humidity conditions for the bioremediation of soil with different concentrations of Cr(VI) and lindane were determined. Lettuce seedlings were a suitable biomarker to evaluate the contaminants mixture toxicity. Streptomyces sp. M7 carried out a successful bioremediation, which was demonstrated through ecotoxicity test with Lactuca sativa.