Construction of a combined soil quality indicator to assess the effect of glyphosate application.

Although the use of agrochemicals allowed increasing the crops productivity, in many cases led to soil deterioration. In this study, eight composite samples from different soils of two locations (San Martín and Anta) in Salta, Argentina, were collected and analyzed. All the samples were from loamy Entisols (0-20 cm depth) under reduced tillage without and with direct spray application of glyphosate. Twenty six variables were determined (physical, chemical, and biological soil quality indicators). From them, those of higher specificity and sensitivity to changes following glyphosate application were identified by a stepwise reduction of variables aided by statistical analysis. Samples were grouped regarding location and application of glyphosate, to identify differential effects upon variables, and glyphosate sensitive variables were selected by discarding those influenced by other factors. Thence, they were used to compose a first approximation to a combined soil quality indicator (CSQI) to assess the effect of glyphosate use in agriculture upon the soil. Overall, the set of physical variables showed the same discriminating structure as the biological set. Finally, two biological, two chemical, and two physical indicators resulted as the most specific to quality variations by the application of the herbicide, being the most sensitive the microbial biomass carbon and the (Aminomethyl)phosphonic acid concentration in soil. When these two were considered into a CSQI, it was possible to discriminate samples with the application of glyphosate (lower quality) from those without application (higher quality). To the best of our knowledge, this is the first attempt to propose a CSQI that could play an important role to prevent degradation in soils subjected to glyphosate application, as it could aid in the early detection of soil quality loss. This would provide to land managers a decision tool to let the land rest from glyphosate application, to ensure sustainable practices in agriculture.

Removal of a mixture of pesticides by a Streptomyces consortium: Influence of different soil systems.

Although the use of organochlorine pesticides (OPs) is restricted or banned in most countries, they continue posing environmental and health concerns, so it is imperative to develop methods for removing them from the environment. This work is aimed to investigate the simultaneous removal of three OPs (lindane, chlordane and methoxychlor) from diverse types of systems by employing a native Streptomyces consortium. In liquid systems, a satisfactory microbial growth was observed accompanied by removal of lindane (40.4%), methoxychlor (99.5%) and chlordane (99.8%). In sterile soil microcosms, the consortium was able to grow without significant differences in the different textured soils (clay silty loam, sandy and loam), both contaminated or not contaminated with the OPs-mixture. The Streptomyces consortium was able to remove all the OPs in sterile soil microcosm (removal order: clay silty loam > loam > sandy). So, clay silty loam soil (CSLS) was selected for next assays. In non-sterile CSLS microcosms, chlordane removal was only about 5%, nonetheless, higher rates was observed for lindane (11%) and methoxychlor (20%). In CSLS slurries, the consortium exhibited similar growth levels, in the presence of or in the absence of the OPs-mixture. Not all pesticides were removed in the same way; the order of pesticide dissipation was: methoxychlor (26%)>lindane (12.5%)>chlordane (10%). The outlines of microbial growth and pesticides removal provide information about using actinobacteria consortium as strategies for bioremediation of OPs-mixture in diverse soil systems. Texture of soils and assay conditions (sterility, slurry formulation) were determining factors influencing the removal of each pesticide of the mixture.

Potential application of a bioemulsifier-producing actinobacterium for treatment of vinasse.

Vinasse is a complex effluent created during production of ethyl alcohol, which can present serious pollution hazard in areas where it is discharged. A variety of technologies, many based upon recovery of the effluent via microbial pathways, are continually being evaluated in order to mitigate the pollution potential of vinasse. The present work reports on initial advances related to the effectiveness of the actinobacterium Streptomyces sp. MC1 for vinasse treatment. Alternative use of raw vinasse as a substrate for producing metabolites of biotechnological interest such as bioemulsifiers, was also evaluated. The strain was able to grow at very high vinasse concentrations (until 50% v/v) and remove over 50% of the biodegradable organic matter in a time period as short as 4 d. Potentially toxic metals such as Mn, Fe, Zn, As, and Pb were also effectively removed during bacterial growth. Decrease in the pollution potential of treated vinasse compared to raw effluent, was reflected in a significant increase in the vigour index of Lactuca sativa (letucce) used as bioremediation indicator. Finally, significant bioemulsifier production was detected when this strain was incubated in a vinasse-based culture medium. These results represent the first advances on the recovery and re-valuation of an actual effluent, by using an actinobacterium from our collection of cultures.

Selection of an actinobacteria mixed culture for chlordane remediation. Pesticide effects on microbial morphology and bioemulsifier production.

Chlordane bioremediation using actinobacteria mixed culture is an attractive clean-up technique. Their ability to produce bioemulsifiers could increase the bioavailability of this pesticide. In order to select a defined actinobacteria mixed culture for chlordane remediation, compatibility assays were performed among six Streptomyces strains. The strains did not show growth inhibition, and they were assayed for chlordane removal, either as pure or as mixed cultures. In pure cultures, all of the strains showed specific dechlorination activity (1.42-24.20 EU mg(-1)) and chlordane removal abilities (91.3-95.5%). The specific dechlorination activity was mainly improved with cultures of three or four microorganisms. The mixed culture consisting of Streptomyces sp. A2-A5-A13 was selected. Their ability to produce bioemulsifiers in the presence of glucose or chlordane was tested, but no significant differences were observed (p > 0.05). However, the stability of the emulsions formed was linked to the carbon source used. Only in chlordane presence the emulsions retained 100% of their initial height. Finally, the selected consortium showed a high degree of sporulation in the pesticide presence. This is the first study on the effects that chlordane exerts on microbe morphology and emulsifier production for a defined mixed culture of Streptomyces with ability to remediate the pesticide.

Removal of the insecticide diazinon from liquid media by free and immobilized Streptomyces sp. isolated from agricultural soil.

From an agricultural soil that had received continuous applications of organophosphorus pesticides, 30 actinobacteria strains were isolated. Two strains, identified as Streptomyces sp. AC1-6 and Streptomyces sp. ISP4, were selected because of their tolerance to diazinon and based on the relationship between diazinon removal and microbial growth. In liquid medium with diazinon at concentrations of 25 and 50 mg L(-1), both strains were able to remove approximately 40-50% and 70-90% of the initial diazinon after 24 and 96 h of incubation, respectively. This diazinon removal was accompanied by microbial growth of the strains, an initial pH decrease, and glucose consumption in the liquid medium. Evaluation of the diazinon removal achieved by the free actinobacteria and Streptomyces sp. AC1-6 immobilized on alginate beads revealed that the immobilized cells exhibited a 60% higher diazinon removal compared with the free cells. The reusability of the encapsulated biomass was confirmed, and a diazinon removal rate of more than 50% was obtained after the second batch. This work constitutes one of the few reports that describe Streptomyces strains as diazinon degraders. Given the high diazinon removal found, the streptomycetes exhibit suitable potential as diazinon-degrading actinobacteria for elimination of diazinon from liquid residues.

Enhanced polyaromatic hydrocarbon degradation by adapted cultures of actinomycete strains.

Fifteen actinomycete strains were evaluated for their potential use in removal of polycyclic aromatic hydrocarbons (PAH). Their capability to degrade of naphthalene, phenanthrene, and pyrene was tested in minimal medium (MM) and MM with glucose as another substrate. Degradation of naphthalene in MM was observed in all isolates at different rates, reaching maximum values near to 76% in some strains of Streptomyces, Rhodococcus sp. 016 and Amycolatopsis tucumanensis DSM 45259. Maximum values of degradation of phenanthrene in MM occurred in cultures of A. tucumanensis DSM 45259 (36.2%) and Streptomyces sp. A12 (20%), while the degradation of pyrene in MM was poor and only significant with Streptomyces sp. A12 (4.3%). Because of the poor performance when growing on phenanthrene and pyrene alone, Rhodococcus sp. 20, Rhodococcus sp. 016, A. tucumanensis DSM 45259, Streptomyces sp. A2, and Streptomyces sp. A12 were challenged to an adaptation schedule of successive cultures on a fresh solid medium supplemented with PAHs, decreasing concentration of glucose in each step. As a result, an enhanced degradation of PAHs by adapted strains was observed in the presence of glucose as co-substrate, without degradation of phenanthrene and pyrene in MM while an increase to up to 50% of degradation was seen with these strains in glucose amended media. An internal fragment of the catA gene, which codes for catechol 1,2-dioxygenase, was amplified from both Rhodococcus strains, showing the potential for degradation of aromatic compounds via salycilate. These results allow us to propose the usefulness of these actinomycete strains for PAH bioremediation in the environment.

Enhanced removal of a pesticides mixture by single cultures and consortia of free and immobilized Streptomyces strains.

Pesticides are normally used to control specific pests and to increase the productivity in crops; as a result, soils are contaminated with mixtures of pesticides. In this work, the ability of Streptomyces strains (either as pure or mixed cultures) to remove pentachlorophenol and chlorpyrifos was studied. The antagonism among the strains and their tolerance to the toxic mixture was evaluated. Results revealed that the strains did not have any antagonistic effects and showed tolerance against the pesticides mixture. In fact, the growth of mixed cultures was significantly higher than in pure cultures. Moreover, a pure culture (Streptomyces sp. A5) and a quadruple culture had the highest pentachlorophenol removal percentages (10.6% and 10.1%, resp.), while Streptomyces sp. M7 presented the best chlorpyrifos removal (99.2%). Mixed culture of all Streptomyces spp. when assayed either as free or immobilized cells showed chlorpyrifos removal percentages of 40.17% and 71.05%, respectively, and for pentachlorophenol 5.24% and 14.72%, respectively, suggesting better removal of both pesticides by using immobilized cells. These results reveal that environments contaminated with mixtures of xenobiotics could be successfully cleaned up by using either free or immobilized cultures of Streptomyces, through in situ or ex situ remediation techniques.

Bacterial bio-resources for remediation of hexachlorocyclohexane.

In the last few decades, highly toxic organic compounds like the organochlorine pesticide (OP) hexachlorocyclohexane (HCH) have been released into the environment. All HCH isomers are acutely toxic to mammals. Although nowadays its use is restricted or completely banned in most countries, it continues posing serious environmental and health concerns. Since HCH toxicity is well known, it is imperative to develop methods to remove it from the environment. Bioremediation technologies, which use microorganisms and/or plants to degrade toxic contaminants, have become the focus of interest. Microorganisms play a significant role in the transformation and degradation of xenobiotic compounds. Many Gram-negative bacteria have been reported to have metabolic abilities to attack HCH. For instance, several Sphingomonas strains have been reported to degrade the pesticide. On the other hand, among Gram-positive microorganisms, actinobacteria have a great potential for biodegradation of organic and inorganic toxic compounds. This review compiles and updates the information available on bacterial removal of HCH, particularly by Streptomyces strains, a prolific genus of actinobacteria. A brief account on the persistence and deleterious effects of these pollutant chemical is also given.

Lindane removal by pure and mixed cultures of immobilized actinobacteria.

Lindane (γ-HCH) is an organochlorine insecticide that has been widely used in developing countries. It is known to persist in the environment and can cause serious health problems. One of the strategies adopted to remove lindane from the environment is bioremediation using microorganisms. Immobilized cells present advantages over free suspended cells, like their high degradation efficiency and protection against toxins. The aims of this work were: (1) To evaluate the ability of Streptomyces strains immobilized in four different matrices to remove lindane, (2) To select the support with optimum lindane removal by pure cultures, (3) To assay the selected support with consortia and (4) To evaluate the reusability of the immobilized cells. Four Streptomyces sp. strains had previously shown their ability to grow in the presence of lindane. Lindane removal by microorganisms immobilized was significantly higher than in free cells. Specifically immobilized cells in cloth sachets showed an improvement of around 25% in lindane removal compared to the abiotic control. Three strains showed significantly higher microbial growth when they were entrapped in silicone tubes. Strains immobilized in PVA-alginate demonstrated lowest growth. Mixed cultures immobilized inside cloth sachets showed no significant enhancement compared to pure cultures, reaching a maximum removal of 81% after 96 h for consortium I, consisting of the four immobilized strains together. Nevertheless, the cells could be reused for two additional cycles of 96 h each, obtaining a maximum removal efficiency of 71.5% when each of the four strains was immobilized in a separate bag (consortium III).

Chromate reductase activity in Streptomyces sp. MC1.

Biological transformation of Cr(VI) to Cr(III) by enzymatic reduction may provide a less costly and more environmentally friendly approach to remediation. In a previous report a Cr(VI) resistant actinomycete strain, Streptomyces sp. MC1, was able to reduce Cr(VI) present in a synthetic medium, soil extract and soil samples. This is the first time optimal conditions such as pH, temperature, growth phase and electron donor have been elucidated in vitro for Cr(VI) reduction by a streptomycete. Chromate reductase of Streptomyces sp. MC1 is a constitutive enzyme which was mainly associated with biomass and required NAD(P)H as an electron donor. It was active over a broad temperature (19-39 degrees C) and pH (5-8) range, and optimum conditions were 30 degrees C and pH 7. The enzyme was present in supernatant, pellet and cell free extract. Bioremediation with the enzyme was observed in non-compatible cell reproduction systems, conditions frequently found in contaminated environments.

Cu(II) removal by Rhodotorula mucilaginosa RCL-11 in sequential batch cultures.

The present study explored the ability of the yeast Rhodotorula mucilaginosa RCL-11 to adapt to increasing Cu(II) concentrations, measuring oxidative stress through superoxide dismutase and catalase activity in two parallel sequential batch assays. One assay was performed in Erlenmeyer flasks without aeration and a second in a fermentor in which the dissolved oxygen was maintained at 30% saturation. Both assays were carried out by increasing Cu(II) concentrations in five sequential steps: 0; 0.1; 0.2; 0.5 and 1 mM. Each assay was incubated at 30 degrees C, 250 rpm and pH 5.5. While growth parameters of R. mucilaginosa RCL-11 decreased 90-95% with increasing Cu(II) concentration in the culture medium, the oxidative stress level increased from 30 to 55% in both assays. Cells grown under controlled oxygen conditions showed 30% more copper bioaccumulation and 10% glucose consumption when compared with cells grown without aeration. SOD activity was higher under controlled than without aeration, whereas CAT activity was similar under both test conditions. Cu(II) bioaccumulation by R. mucilaginosa RCL-11 and a possible increase in this capacity by adaptation of the strain under controlled aeration represent a potential valuable tool for treatment of effluents or water bioremediation with high copper contents.

Responses of Candida fukuyamaensis RCL-3 and Rhodotorula mucilaginosa RCL-11 to copper stress.

The effect of high Cu(II) concentrations on superoxide dismutase (SOD) and catalase (CAT) activity in Candida fukuyamaensis RCL-3 and Rhodotorula mucilaginosa RCL-11, previously isolated from a copper filter at a mine plant in Argentina, was studied. Addition of 0.1, 0.2 and 0.5 mM Cu(II) to the culture medium increased total SOD and CAT activity in both strains. Native polyacrylamide gel electrophoresis revealed two bands with SOD activity for C. fukuyamaensis RCL-3 and only one for R. mucilaginosa RCL-11; the three bands corresponded to MnSOD.Intracellular accumulation of copper and morphological changes was observed using electron microscopy. Dark bodies examined with transmission electron microscopy (TEM) after 48 h of incubation probably corresponded to copper deposits. The number of dark bodies in R. mucilaginosa RCL-11 grew with increasing incubation time, whereas in C. fukuyamaensis RCL-3 the amount decreased. Scanning electron micrographs (SEM) of C. fukuyamaensis RCL-3 did not reveal any differences compared with the control, but R. mucilaginosa RCL-11 cells were bigger than control ones. TEM confirmed absence of compartmentalization mechanisms in Cu(II) detoxification since electron-dense bodies were mainly found in the cytoplasm.

Bioremediation of chromium(VI) contaminated soil by Streptomyces sp. MC1.

This work provides quantitative information on Cr(VI) reduction in soil samples by an indigenous actinomycete. Streptomyces sp. MC1, previously isolated from sugarcane, has shown ability to reduce Cr(VI) in liquid minimal medium. A reduction of 100 and 75% was obtained at initial Cr(VI) concentrations of 5 and 50 mg l(-1), respectively, after 48 h of incubation. Bioremediation ability of Streptomyces sp. MC1 was assayed in soil extracts and soil samples. Relative growth of Streptomyces sp. MC1 was 77 and 38% when grown in soil extract with 10 and 50 mg l(-1) of Cr(VI), respectively. MC1 was able to reduce 30% of Cr(VI) after 96 h of incubation with 10 mg l(-1) of Cr(VI), and reduction coincided with the exponential growth phase at pH 7 and 30 degrees C.In soil samples, Streptomyces sp. MC1 was able to reduce up to 94% of the Cr(VI) bioavailability (50 mg kg(-1)) after 7 d. These results were compared with non-inoculated soil samples with Cr(VI). Bioremediation activity of Streptomyces sp. MC1 was not inhibited by natural soil microbial flora. Besides, Streptomyces sp. MC1 growth was not inhibited by 50 mg kg(-1) of Cr(VI). In contrast to findings obtained by other authors, our results showed almost complete Cr(VI) removal from soil without any previous treatment, and without addition of any substrate and with a normal soil humidity level. These results confirm the Cr(VI)-contaminated soil bioremediation potential of Streptomyces sp. MC1.

Chromate removal by yeasts isolated from sediments of a tanning factory and a mine site in Argentina.

Twenty-one yeast-like microorganisms were isolated from tannery effluents and from a nickel-copper mine in Argentina. They were tested for their Cu(II), Ni(II), Cd(II) and Cr(VI) tolerance in qualitative assays on solid medium. Three isolates were selected for their multiple tolerance to the different heavy metals and highest tolerance to Cr(VI). According to morphological and physiological analysis and 26S rDNA D1/D2 domain sequences the isolates were characterized as: Lecythophora sp. NGV-1, Candida sp. NGV-9 and Aureobasidium pullulans VR-8. Resistance of the three strains to high Cr(VI) concentrations and their ability to remove Cr(VI) were assessed using YNB-glucose medium supplemented with 0.5 and 1 mM Cr(VI). Chromate removal activity was estimated by measuring remaining Cr(VI) concentration in the supernatant using the colorimetric 1,5-diphenylcarbazide method and total chromium was determined by flame atomic absorption spectroscopy. The results indicate that the initial Cr(VI) concentration negatively influenced growth and the specific growth rate but stimulated the metabolic activity of the three strains; resistance to Cr(VI) by these strains was mainly due to reduction of Cr(VI) rather than chromium bioaccumulation. This study showed the potential ability of these strains as tools for bioremediation of Cr(VI) from contaminated sites.

Chromium(VI) resistance and removal by actinomycete strains isolated from sediments.

Forty-one isolated actinomycetes were used to study qualitative and semi-quantitative screening of chromium(VI) resistance. Chromate-removing activity was estimated using the Cr(VI) specific colorimetric reagent 1,5-diphenylcarbazide. Twenty percent of the isolates from El Cadillal (EC) and 14% of isolates from a copper filter plant (CFP) were able to grow at 13 mM of Cr(VI). All isolates from sugar cane (SCP) could grow up to Cr(VI) concentration of 17 mM. EC, CFP and SCP strains were able to remove 24%, 30% and more than 40% of Cr(VI), respectively. The highest and lowest Cr(VI) specific removal values were 75.5 mg g(-1) cell by M3 (CFP), and 1.5 mg g(-1) cell by C35 (EC) strains. Eleven Cr(VI) resistant strains were characterized and identified as species of the genera Streptomyces (10) and Amycolatopsis (1). Differences on actinomycete community composition between contaminated and non-contaminated soil were found. This study showed the potential capacity of actinomycetes as tools for Cr(VI) bioremediation.

Copper tolerant yeasts isolated from polluted area of Argentina.

Eleven yeasts were isolated from wastewater sediment samples collected from a copper filter mine plant, located in the province of Tucumán, Argentina, and tested for their heavy metal tolerance. Two isolates were selected based on their multiple tolerance to different heavy metals and their copper biosorption capacity was studied. Analysis of the 26S rDNA D1/D2 domain sequences indicates that isolate RCL-3 showed similarity with Candida sp. and RCL-11 with Rhodotorula mucilaginosa . Growth performance and copper toxicity of both yeasts were evaluated using YNB-glucose medium supplemented with 0.1, 0.2, 0.5 and 1 mM of Cu2+ solutions. Candida sp. RCL-3 was able to grow up to 7 mg ml(-1) biomass in the presence of either 0.1 or 0.2 mM Cu2+, and at 0.5 mM Cu2+ growth reached 5.5 mg ml(-1). R. mucilaginosa RCL-11 reached 8 mg ml(-1) in the presence of 0.1 mM Cu2+, and values of 6.5 and 5.5 mg ml(-1) biomass were obtained at 0.2 and 0.5 mM Cu2+, respectively. Copper accumulation profiles were different: the metal was librated from the intact cells by Candida sp. whereas R. mucilaginosa did not show release from the cells indicating intracellular storage. Specific biosorption of copper by both isolated yeasts showed increase with the initial copper supplied with the medium (up to 11.5 and 8.0 mg Cu g(-1) biomass for Candida sp. and R. mucilaginosa , respectively). However, specific biosorption decreased with time.

Estimation of growth inhibition by copper and cadmium in heavy metal tolerant actinomycetes.

A new isothermic model of actinomycetes growth in presence of toxic concentrations of Cd(2+) and Cu(2+) is described. Microbial growth inhibition, displayed as a decrease of biomass, can be correlated with the increase of cadmium(II) or copper(II) concentration in the medium. The reciprocal dry biomass against metal concentration showed a linear correlation higher than 0.9 in tested strains. The mathematical model can be useful to predict the behavior of actinomycetes at inhibitory concentrations of copper(II) and cadmium(II) in large screening procedures.

Screening of heavy metal-tolerant actinomycetes isolated from the Salí River.

Fifty-three strains of actinomycetes resistant to heavy metals were isolated from the Salí River in northwest Argentina. Screening procedures that involve solid and liquid synthetic media containing Cd(2+), Cu(2+), or Hg(2+) allowed the selection of six strains. These strains showed a quantitative sorption of Cd(2+) and Cu(2+) by more than 98% of the initial metal concentrations (0.1, 0.5, and 1.0 mM) tested.

Estudio de la dinámica de la carga orgánica, bacteriana y viral en el Embalse El Cadillal

El Embalse El Cadillal es el más importante de la Pcia. de Tucumán ya que abastece de agua potable al 70o/o de la población. Sus aguas son permanentemente analizadas para determinar el nivel de eutroficación. El objetivo de este trabajo es emplear indicadores no convencionales para incluir en el estudio de las aguas del embalse: dosaje de actinomicetes y recuperación de virus entéricos