Microbial diversity and abundance of Hg related genes from water, sediment and soil the Colombian amazon ecosystems impacted by artisanal and small-scale gold mining.

The Amazon region abounds in precious mineral resources including gold, copper, iron, and coltan. Artisanal and small-scale gold mining (ASGM) poses a severe risk in this area due to considerable mercury release into the surrounding ecosystems. Nonetheless, the impact of mercury on both the overall microbiota and the microbial populations involved in mercury transformation is not well understood. In this study we evaluated microbial diversity in samples of soil, sediment and water potentially associated with mercury contamination in two localities (Taraira and Tarapacá) in the Colombian Amazon Forest. To this end, we characterized the bacterial community structure and mercury-related functions in samples from sites with a chronic history of mercury contamination which today have different levels of total mercury content. We also determined mercury bioavailability and mobility in the samples with the highest THg and MeHg levels (up to 43.34 and 0.049 mg kg-1, respectively, in Taraira). Our analysis of mercury speciation showed that the immobile form of mercury predominated in soils and sediments, probably rendering it unavailable to microorganisms. Despite its long-term presence, mercury did not appear to alter the microbial community structure or composition, which was primarily shaped by environmental and physicochemical factors. However, an increase in the relative abundance of merA genes was detected in polluted sediments from Taraira. Several Hg-responsive taxa in soil and sediments were detected in sites with high levels of THg, including members of the Proteobacteria, Acidobacteria, Actinobacteria, Firmicutes and Chloroflexi phyla. The results suggest that mercury contamination at the two locations sampled may select mercury-adapted bacteria carrying the merA gene that could be used in bioremediation processes for the region.

Highly mercury-resistant strains from different Colombian Amazon ecosystems affected by artisanal gold mining activities.

Two sites of the Colombian Amazon region with different levels of human intervention and mercury pollution were selected for the collection of samples of river and lake water, sediments, and associated forest soils. The Tarapacá region, affected mainly by barrage mining, showed low mercury concentrations, whilst in the Taraira region, affected by underground mining, there were several points with high mercury pollution levels. A collection of 72 bacterial and 10 yeast strains with different levels of mercury resistance was isolated and characterized. Most of the highly resistant bacterial strains (MIC > 40 mg L-1 HgCl2) were isolated from soil and sediment samples and belonged to either Pseudomonas (60%) or Bacillus (20%). Most of highly resistant bacterial strains were positive for the presence of the merA gene, suggesting an active mercury resistance mechanism. This was confirmed in the two most resistant strains, Pseudomonas sp. TP30 and Burkholderia contaminans TR100 (MIC = 64 and 71 mg L-1 HgCl2, respectively), which in the presence of increasing mercury concentrations expressed the merA gene at increasing levels, concomitant with a significant mercury reduction activity. Analysis of the MerA sequences present in the different isolates suggested a high gene conservation within the taxonomic groups but also several horizontal gene transfer events between taxonomically distant genera. We also observed a positive correspondence between the presence of the merA gene and the number of antibiotics to which the strains were resistant to. The most resistant strains are good candidates for future applications in the bioremediation of mercury-contaminated sites in the Amazon.Key points• Amazon sediments affected by underground gold mining have higher Hg levels.• Highly Hg-resistant isolates belonged to Pseudomonas and Bacillus genera.• TR100 and TP30 strains showed remediation potential to be used in the Amazon region.

Comunidades de hongos actinomicetos en tres tipos de vegetación de la Amazonia colombiana: abundancia, morfotipos y el gen 16s ADNr

Communities of Actynomicetes fungy in three vegetation types of the Colombian Amazon: abundance, morphotypes and the 16s rDNA gene. Among soil microorganisms, Actinomycetes play an important role in the sustainability of natural and agricultural systems: decomposition of organic matter; degradation of recalcitrant compounds like lignin; nitrogen fixation; degradation of agricultural chemicals and biological control in plants and animals. We evaluated their diversity in soils under three different vegetation covers (pasture, tropical primary forest and stubble) at two depths in the Southern Colombian Amazon border. We collected five replicates per vegetation type (in each, three samples at 0-20cm and three at 20-30cm; for a total of 30 samples). Abundance and phenotypic diversity were determined by plate counting. Genomic DNA was extracted from the isolates: the 16s rDNA gene was amplified with specific primers, and its genetic diversity was estimated by means of an amplified restriction analysis (ARDRA). Actynomicetes abundance varied with vegetation and depth, possibly reflecting presence of earthworms, macro-fauna and physico-chemical characteristics associated to fertility, as well as organic matter, total bases, and optimal capacity to cationic interchange. Primary forests had the highest diversity. Sixteen morpho-types (six genera) were identified; Streptomyces was the most abundant everywhere. The heterogeneity of ARDRA patterns prevented species identification because of the intra-species variability in sequences of 16s rDNA operons. This community is a biological indicator of landscape alteration and could include new bio-active compounds of pharmaceutical interest. Rev. Biol. Trop. 57 (4): 1119-1139. Epub 2009 December 01.

Los actinomicetos son importantes en la sostenibilidad de sistemas naturales. Su diversidad fue evaluada en suelos de bosque, pastizal y rastrojo, y dos profundidades en el Sur del Trapecio Amazónico Colombiano. Se analizaron suelos de cinco repeticiones por cobertura para un total de 15 unidades. Se tomaron seis muestras en cada unidad y dos profundidades, para un total de 30. Los actinomicetos cultivables se determinaron por recuento en placa, se extrajo ADN, se amplificó el gen ADNr 16s y su diversidad genética se estimó por ARDRA. Hubo diferencias de abundancia entre coberturas y profundidades, relacionadas con la vegetación, presencia de lombrices, macrofauna, altos niveles de materia orgánica, y bases totales. Se obtuvieron valores de diversidad fenotípica similares para las tres coberturas, pero los bosques son más diversos. Se identificaron 16 morfotipos, agrupados en séis géneros, siendo Streptomyces el más abundante. La heterogeneidad de los patrones ARDRA no permitió la asignación de especies, reflejándose variaciones en las secuencias de diferentes operones ADNr 16s en un mismo organismo. Las perturbaciones en la cobertura influyen sobre los actinomicetos, generando cambios en su abundancia y diversidad. Su importancia ecológica permite proponerlos como indicadores biológicos de alteración del paisaje.

[Communities of Actynomicetes fungy in three vegetation types of the Colombian Amazon: abundance, morphotypes and the 16s rDNA gene]. / Comunidades de hongos actinomicetos en tres tipos de vegetación de la Amazonia colombiana: abundancia, morfotipos y el gen 16s ADNr.

Among soil microorganisms, Actinomycetes play an important role in the sustainability of natural and agricultural systems: decomposition of organic matter; degradation of recalcitrant compounds like lignin; nitrogen fixation; degradation of agricultural chemicals and biological control in plants and animals. We evaluated their diversity in soils under three different vegetation covers (pasture, tropical primary forest and stubble) at two depths in the Southern Colombian Amazon border. We collected five replicates per vegetation type (in each, three samples at 0-20cm and three at 20-30cm; for a total of 30 samples). Abundance and phenotypic diversity were determined by plate counting. Genomic DNA was extracted from the isolates: the 16s rDNA gene was amplified with specific primers, and its genetic diversity was estimated by means of an amplified restriction analysis (ARDRA). Actynomicetes abundance varied with vegetation and depth, possibly reflecting presence of earthworms, macro-fauna and physico-chemical characteristics associated to fertility, as well as organic matter, total bases, and optimal capacity to cationic interchange. Primary forests had the highest diversity. Sixteen morpho-types (six genera) were identified; Streptomyces was the most abundant everywhere. The heterogeneity ofARDRA patterns prevented species identification because of the intra-species variability in sequences of 16s rDNA operons. This community is a biological indicator of landscape alteration and could include new bio-active compounds of pharmaceutical interest.

Micorrizas arbusculares del sur de la Amazonia colombiana y su relación con algunos factores fisicoquímicos y biológicos del suelo / x

La presencia de micorrizas arbusculares en la vegetación de la Amazonia mejora la nutrición de las plantas en suelos de baja fertilidad. Este trabajo evaluó la presencia natural de hongos micorrícicos de tipo arbuscular (HMA) en suelos ácidos de textura franco-arcillosa a arcillosa del sur de la Amazonia colombiana bajo bosque, rastrojo joven, y praderas establecidas, a dos profundidades diferentes. Fue estudiada la presencia de HMA (riqueza y abundancia de esporas) relacionado con la acidez, la capacidad de intercambio catiónico, el carbono orgánico, el fósforo total, las fracciones de fósforo soluble y fijado al aluminio, hierro y calcio, y el ADN total del suelo. Se detectaron diferencias significativas, en el contenido de ADN total y el número de esporas respecto a la profundidad de muestreo. El pH presentó un efecto significativo sobre el contenido de ADN y el número de esporas de HMA. El contenido de ADN en el suelo se vio afectado por las concentraciones de fosfatos de aluminio, mientras la esporulación de HMA fue afectada por las concentraciones de fosfatos de hierro del suelo. Así, el número de esporas de HMA en suelos de la Amazonia se ve afectada por la profundidad, el pH y por el tipo de fosfatos minerales presentes.

A presença das micorrizas arbusculares na vegetação da Amazônia pode incrementar a nutrição das plantas nos solos com baixa fertilidade. Neste estudo foi avaliada a ocorrência natural de fungos formadores de micorriza arbuscular (MA) em solos ácidos de textura argilosa no sul da Amazônia colombiana, considerando-se nas coberturas de floresta nativa, floresta secundaria jovem, e pastagens estabelecidas e duas profundidades. Foi estudada a ocorrência das MA (riqueza e abundância de esporos) com relação à: acidez, capacidade de troca de cátions, carbono orgânico, fósforo total, frações de fósforo solúvel, frações fixadas ao alumínio, ferro e cálcio e DNA total do solo. Quanto à profundidade, encontraram-se diferenças significativas, tanto para o conteúdo de DNA total e ao número de esporos. Entre as variáveis químicas o pH teve efeito significativo sob o conteúdo de DNA e o número de esporos dos fungos MA. O conteúdo de DNA foi influenciado pelas concentrações de fosfatos de alumínio do solo, em quanto a esporulação de fungos MA foi influenciada pelas concentrações de fosfatos de ferro. Concluiu-se que a abundância dos fungos MA variou dependendo da profundidade do estrato avaliada, o pH e os fosfatos minerais presentes no solo.