Sensitivity of microbial bioindicators in assessing metal immobilization success in smelter-impacted soils.

While plant toxicity reduction remains the primary metric for judging the success of metal immobilization in soil, the suitability of microorganisms as universal indicators of its effectiveness in various contaminated soils remains a point of contention. This study assessed the sensitivity of microbial bioindicators in monitoring metal immobilization success in smelter-impacted soils. It compared plants and microorganisms as indicators of the efficiency of natural Fe-Mn nodules from the Gulf of Finland in immobilizing metals in soils contaminated by a Ni/Cu smelter, on the Kola Peninsula, Murmansk region, Russia. Perennial ryegrass (Lolium perenne) was grown on nodule-amended and control soils. Plant responses in the smelter-impacted soils proved to be sensitive and robust indicators of successful metal immobilization. However, microbial responses exhibited a more complex story. Despite the observed reductions in soluble metal concentrations, shoot metal contents in ryegrass, and significant improvements in plant growth, certain microbial bioindicators were unresponsive to metal immobilization success brought about by the addition of Fe-Mn nodules. Among microbial bioindicators studied, community-level physiological profiling, microbial biomass carbon, and basal respiration were sensitive indicators of metal immobilization success, whereas the number of saprotrophic, oligotrophic, and Fe-oxidizing bacteria and fungi, the biomass of bacteria and fungi, and enzymatic activity were less robust indicators. Interestingly, the correlations between different microbial responses measured were weak or even negative. Some microbial responses also exhibited negative correlations with plant biomass. These findings underscore the need for further research on comparative evaluations of plants and microorganisms as reliable indicators of metal immobilization efficacy in polluted environments.

Zinc shields against copper phytotoxicity in a contaminated soil.

While zinc protects plants from copper in hydroponics, its behavior in soil remains unclear. We investigated the potential of zinc sulfate to protect ryegrass from copper toxicity in contaminated soil. Twelve soil treatments combined varying levels of copper oxide (CuO) and zinc sulfate (ZnSO4). Increasing CuO significantly stunted ryegrass, but adding ZnSO4 mitigated the effects at each CuO level. ZnSO4 had no effect in unpolluted conditions. These results, supported by the Terrestrial Biotic Ligand Model, indicate that zinc competes with copper for binding sites, reducing copper uptake by ryegrass and mitigating its toxicity. Application of zinc sulfate to copper-contaminated soils appears promising for ryegrass growth, although field studies are critical to confirm real-world efficacy.

The downside of copper pesticides: An earthworm's perspective.

The widespread use of copper-based pesticides, while effective in controlling plant diseases, has been identified as a major source of copper contamination in soils. This raises concerns about potential adverse effects on earthworms, key players in soil health and ecosystem function. To inform sustainable pesticide practices, this study aimed to establish copper toxicity thresholds for earthworm avoidance in agricultural soils impacted by copper-based pesticides. We collected 40 topsoil samples (0-5 cm) from orchards and vineyards in the O'Higgins Region of central Chile, and 10 additional soils under native vegetation as background references. A standardized avoidance bioassay using Eisenia fetida assessed the impact of copper-based pesticides on the soils. Total copper concentrations ranged between 23 and 566 mg kg-1, with observed toxic effects on earthworms in certain soils. The effective concentration at 50% (EC50) for total soil copper, determined by Eisenia fetida's avoidance response, was 240 mg kg-1, with a 95% confidence interval of 193-341 mg kg-1. We further compared our EC50 values with existing data from agricultural soils impacted by mining activities. Interestingly, the results revealed a remarkable similarity between the thresholds for earthworm avoidance, regardless of the source of copper contamination. This observation underscores the universality of copper toxicity in agricultural ecosystems and its potential impact on soil biota. This study provides novel insights into copper toxicity thresholds for earthworms in real-world, pesticide-contaminated soils.

First Report of Pantoea ananatis and Pantoea eucalypti causing onion leaf blight and bulb decay in Central Chile.

Nearly 5,400 hectares of long-day onions (Allium cepa) are cultivated in the Central Zone of Chile (UTM 33°3´S to 37°24´S). During summer 2021-22 and 2022-23 (December to February), followed by high temperatures (around 35°C) and high humidity conditions, symptoms showing yellowing, soft rot, blight on old leaves, necrotic tips, and soft bulbs were observed. The affected plants were observed in "spots" into the fields, and the incidence reached 5-10%. The severity was high, and 70-80% of the affected plants died. Symptomatic plants from different fields from the Coquimbo, Metropolitan, O´Higgins and Maule regions were sampled. Isolations were made using casamino-acid peptone glucose (CPG) agar medium (Schaad, 2001). Yellow-pigmented, circular to irregular shaped colonies were observed. Molecular identification was carried out by partial 16S rRNA gene sequencing, resulting in the identification of 18 isolates of Pantoea spp. from twelve different fields. Selected strains were biochemically analyzed using the GEN III BIOLOG microtest system (Hayward, CA) and were identified as Pantoea spp. BLAST analyses of the 16S rRNA sequences (602 nt) of selected strains (GenBank Accession No. OR527817 to OR527819) against the NCBI Database resulted in the identification of Pantoea species with 100% coverage and 100% identity. To determine the Pantoea species of each strain, housekeeping gene gyrB (Delétoile et al. 2009) was amplified and sequenced (GenBank Accessions No. OR544061 to OR544063). BLAST analysis (802 nt) of selected strains resulted in 100% coverage and 100% identity, identifying three different species: P. ananatis, P. eucalypti, and P. agglomerans. Pantoea species were isolated from both leaves and bulbs and no more than one species was observed per field. Pathogenicity assays in onion plants and bulbs were performed based on the methodology described by Asselin et al. (2018). Five onion plants cv. Cimarron of thirteen-week-old were inoculated by wounding an external leave with a sterile toothpick previously immersed in a bacterial suspension at ~ 108 CFU/mL and maintained at 26-28°C for 30 days in high humidity conditions. Control plants were inoculated with sterile water. Plants inoculated with Pantoea spp. showed chlorosis, soft rot, and necrosis mainly in older leaves, as observed in the field, while negative control plants remained healthy. Pantoea spp. were re-isolated from the inoculated onion plants. Toothpicks dipped in the inoculum were stuck 4 cm into the shoulders of onion bulbs and incubated at 26°C for 20 days. Water was inoculated as a negative control. At the end of the incubation period, the bulbs were opened longitudinally across their inoculation sites and shrunken, brownish, watery scales were observed. Pantoea species have been previously described as causing leaf blight of onions in Georgia and Michigan in the USA and South Africa, Brazil, and Uruguay (Hattingh and Walters 1981; Gitaitis and Gay 1997; Edens et al. 2006; Tho et al. 2015; De Armas et al. 2022; Rosende et al. 2022). P. agglomerans was recently reported in Chile (Sepúlveda et al. 2023), but this is the first report of P. ananatis and P. eucalypti affecting onions in central Chile. This detection is an alert call for the onion's growers and exporters in Chile for upcoming seasons, where conditions predisposing to disease may continue to occur. It is crucial to continue analyzing the factors that caused the appearance of this new disease in onions.

Extremely High Soil Copper Content, yet Low Phytotoxicity: A Unique Case of Monometallic Soil Pollution at Kargaly, Russia.

Ecotoxicological studies on soil metal toxicity often rely on artificially contaminated soils. A major difficulty in using soils contaminated by anthropogenic activities (e.g., mining and agriculture) is the presence of multiple metals, which can make it impossible to distinguish the toxic effects of a particular metal. Therefore, sites with monometallic pollution have great potential for ecotoxicological research. One such site is an agricultural field in Kargaly, Orenburg region, Russia, where copper mining and smelting activities were carried out during the 18th-20th centuries. Samples of Mollisols (chernozems) were collected in the studied field. At several sampling points there were copper ore rocks on the surface, containing malachite (CuCO3 · Cu(OH)2 ). The soil samples had a high copper content, up to approximately 10 g kg-1 , compared with 75 mg kg-1 in the background soil. Importantly, the content of other elements in all soil samples was similar to that in the background soil, highlighting the uniqueness of the monometallic contamination in the study area. Despite the extremely high total copper content, exchangeable copper was relatively low, with a maximum of approximately 0.5 mg kg-1 . We performed a short-term (21-day) ecotoxicity assessment using perennial ryegrass as an indicator of copper toxicity. Contrary to expectations, plant growth was not affected by the high copper content in the studied soils. The low copper phytotoxicity may be explained by the low solubility of malachite. However, future long-term experiments may be warranted to determine copper toxicity thresholds for plants under field conditions. The site discovered in the present study could potentially acquire the same significance as the Danish Hygum site for the study of monometallic soil contamination. Environ Toxicol Chem 2023;42:707-713. © 2023 SETAC.

An Assessment of the Feasibility of Phytoextraction for the Stripping of Bioavailable Metals from Contaminated Soils.

Phytoextraction has been proposed in many papers as a low-cost method for remediating contaminated soil. However, if national regulation is based on total metal(loid) concentrations in soil, phytoextraction is generally infeasible because of the long time required for remediation. Assessing phytoextraction requires determination of the dynamic rate of metal removal from soil. Phytoextraction may be feasible if the main goal is to reduce the soluble fraction of the metal(loid) with the goal of reducing bioavailability. However, it has been reported that there is a large mass balance mismatch between the reduction of the soluble metal fraction in contaminated soil and metal uptake by plants. Several studies report that the decrease of soluble fraction of metals in soil is higher than can be accounted for by plant uptake. In other words, studies generally overestimate the feasibility of bioavailable contaminant stripping. Therefore, a more rigorous approach is advisable to ensure that papers on bioavailable contaminant stripping include relevant information on mass balances. Furthermore, to implement the concept of bioavailable contaminant stripping, regulations must distinguish between the bioavailable fraction and the total metal concentration in soil. Environ Toxicol Chem 2023;42:558-565. © 2022 SETAC.

Estudio de calidad de vida y comportamiento adaptativo en niños y adolescentes con discapacidad intelectual / Study of Quality of Life and Adaptive Behavior in Children and Adolescents with Intellectual Disability

Introducción: Los niños y adolescentes con discapacidad intelectual (DI) requieren de evaluaciones cognitivas, adaptativas y de calidad de vida (CV) con el fin de programar estrategias integrales de intervención basadas en sus necesidades. El objetivo de este estudio es describir CV, comportamiento adaptativo y cognición en una serie de niños y adolescentes con DI. Método: Se estudiaron 28 pacientes entre 6 a 18 años con escala de CV, evaluaciones cognitivas y comportamiento adaptativo. Resultados: En escala de CV se obtuvo una puntuación promedio, rango percentil 45-50, con menor puntaje en dimensiones de desarrollo personal, relaciones interpersonales e inclusión social. En escala de comportamiento adaptativo la mayoría de los pacientes presentaron nivel adaptativo bajo, sus dominios más afectados fueron comunicación y socialización. Al relacionar CV, comportamiento adaptativo y cognición se encontró una correlación significativa entre función adaptativa general y cognición (r = ,74, p < ,01) y entre función adaptativa e índice de calidad de vida (r = ,63, p < ,01). Conclusiones: En nuestra serie de niños y adolescentes con DI se relaciona un menor comportamiento adaptativo con menor CV y menor cognición. Inclusión social, desarrollo personal y relaciones interpersonales, así como socialización y comunicación, son las líneas a considerar como planes de intervención. (AU)

Children and adolescents with intellectual disabilities (ID) require cognitive, adaptive and quality of life (QoL) assessments in order to program integral strategies of intervention based on their needs. The objective of this study is to describe quality of life, adaptive behavior and cognition in a series of children and adolescents with ID. Method: 28 patients between 6 and 18 years old were studied with QoL scales, adaptive behavior and cognitive evaluations, and their correlations. Results: On the CV scale, an average score was obtained, 45-50 percentile range, with a lower score in dimensions of personal development, interpersonal relationships and social in-clusion. On the adaptive behavior scale, most of the patients presented a low adaptive level; their most affected domains were communication and socialization. When relat-ing QoL, adaptive behavior and cognition, a significant correlation was found between general adaptive function and cognition (r = ,74, p < ,01) and between adaptive function and quality of life index (r = ,63, p < ,01). Conclusions: In our series of children and adolescent with ID, a lower adaptive behavior is associated with a lower QoL and low-er cognition. Social inclusion, personal development and interpersonal relationships, as well as socialization and communication, are the lines to consider as intervention plans. (AU)

Microbial Community-Level Physiological Profiles and Genetic Prokaryotic Structure of Burned Soils Under Mediterranean Sclerophyll Forests in Central Chile.

Forest fires alter soil microbial communities that are essential to support ecosystem recovery following land burning. These alterations have different responses according to soil abiotic pre- and post-fire conditions and fire severity, among others, and tend to decrease along vegetation recovery over time. Thus, understanding the effects of fires on microbial soil communities is critical to evaluate ecosystem resilience and restoration strategies in fire-prone ecosystems. We studied the state of community-level physiological profiles (CLPPs) and the prokaryotic community structure of rhizosphere and bulk soils from two fire-affected sclerophyll forests (one surveyed 17 months and the other 33 months after fire occurrence) in the Mediterranean climate zone of central Chile. Increases in catabolic activity (by average well color development of CLPPs), especially in the rhizosphere as compared with the bulk soil, were observed in the most recently affected site only. Legacy of land burning was still clearly shaping soil prokaryote community structure, as shown by quantitative PCR (qPCR) and Illumina MiSeq sequencing of the V4 region of the 16S rRNA gene, particularly in the most recent fire-affected site. The qPCR copy numbers and alpha diversity indexes (Shannon and Pielou's evenness) of sequencing data decreased in burned soils at both locations. Beta diversity analyses showed dissimilarity of prokaryote communities at both study sites according to fire occurrence, and NO3 - was the common variable explaining community changes for both of them. Acidobacteria and Rokubacteria phyla significantly decreased in burned soils at both locations, while Firmicutes and Actinobacteria increased. These findings provide a better understanding of the resilience of soil prokaryote communities and their physiological conditions in Mediterranean forests of central Chile following different time periods after fire, conditions that likely influence the ecological processes taking place during recovery of fire-affected ecosystems.

Microbial responses are unreliable indicators of copper ecotoxicity in soils contaminated by mining activities.

Metal contamination of soil has become a serious environmental problem worldwide. Many studies have attempted to infer metal ecotoxicity from soil microbial responses. However, much of the data from these studies tends to be inconsistent and difficult to interpret. We hypothesized that microbial response would be a useful indicator of metal toxicity in soils contaminated by copper mining in Chile. Eighty-four topsoils (0-20 cm) were collected from three areas historically contaminated by copper mining (total Cu: 46-1106 mg kg-1, soluble Cu: 0.05-2.3 mg kg-1, pCu2+: 6.3-10, pH: 5.1-7.8, organic matter: 1.1-10%, clay: 0-28%). Based on soil metal concentrations and ecotoxicity thresholds, Cu was expected to be toxic to microorganisms in the studied soils, while the effects of other metals (total Zn: 79-672, As: 1.9-60, Pb: 19-220, Cd: 0.4-5.1 mg kg-1) were expected to be minor. Soil microbial responses (microbial biomass and numbers, nitrogen mineralization and nitrification, and community-level physiological profiles) were also measured. The results showed that the different responses of soil microbes were not correlated with each other. Furthermore, the soil microbial responses were mainly influenced by the physicochemical properties of the soil, not by the metal concentrations in the soil. The effect of copper on the microbial response was either stimulating (positive) or toxic (negative). Of the soil microbial responses measured in this study, only the microbial biomass was useful for calculating dose-response curves. However, the microbial biomass response was not consistent among the different soil copper pools (total copper, soluble copper, and activity of free Cu2+ ions). It is important to emphasize that the thresholds obtained for copper toxicity cannot be adopted in a robust manner because of the different microbial responses in different sampling areas. Thus, in the copper-contaminated soils under study, microbial response was found to be an unreliable indicator of metal toxicity.

[Prevalence estimation of Autism Spectrum disorders in chilean urban population]. / Estimación de la prevalencia de trastorno del Espectro Autista en población urbana chilena.

INTRODUCTION: The prevalence of Autism Spectrum Disorder has increased, varying between 0.5 and 1% around the world. The prevalence of ASD in Chile is unknown. OBJECTIVE: To estimate the prevalence of ASD in two urban communes of Santiago, Chile. SUBJECTS AND METHOD: Cross-sectional epidemiological study. 272 children aged between 18-30 months who attended well-child visits at two Family Health Centers in two urban communes of Santiago participated. Consecutive sampling was used and chil dren who were already being monitored by neurology were excluded. Screening was performed using the Modified Checklist for Autism in Toddlers (M-CHAT). Those children with altered M-CHAT were evaluated by a pediatric neurologist at the San Borja Arriarán Clinical Hospital and diagnosed with ASD according to clinical criteria. The Autism Diagnostic Observation Schedule - Second Ver sion (ADOS-2) was used as a diagnostic complement. The prevalence of ASD was estimated with a 95% confidence interval. RESULTS: 44 children had altered M-CHAT; 5 of them were clinically diagno sed with ASD. A 1.95% prevalence of ASD (95% CI 0.81-4.63) was obtained, with a sex distribution of 4 boys per 1 girl. CONCLUSIONS: This study is the first estimate of ASD prevalence in two communes of Santiago, Chile. A high prevalence of this condition was observed, which highlights the need for obtaining resources for an early multidisciplinary approach for these patients.

Gypsum soil amendment in metal-polluted soils-an added environmental hazard.

Scientists around the world have long been searching for effective strategies to reduce the bioavailability of metals in contaminated soils. In case of metal-spiked soils, some studies have proposed gypsum as a soil amendment to alleviate metal phytotoxicity. However, for real field-collected soils, evidence on the efficacy of gypsum as a metal phytotoxicity amendment is limited. Therefore, the present study was designed to examine the effect of gypsum on plant growth in soils polluted by a copper smelter. We grew perennial ryegrass on untreated and gypsum-treated soils (at a dose of 3% by weight) under laboratory conditions. We found that gypsum had no effect on alleviating metal phytotoxicity in our soils. We also demonstrated - for the first time - that gypsum increased the concentrations of soluble metals in the soil, enhancing metal uptake by plants. The calcium ions from gypsum displace metals in the soil exchangeable complex; however, the metals do not get immobilized in soils because gypsum is a neutral salt. While our results contrast with the Terrestrial Biotic Ligand Model, that Model has never been tested on real industrially polluted soils but only on metal-spiked soils. Our main conclusion is that gypsum is ineffective in alleviating metal phytotoxicity in real industrially polluted soils and, moreover, its use is inappropriate as a soil remediation method, because it increases the environmental hazard rather than reducing it. Our study is the very first attempt to recognize that gypsum is a hazardous material when used to ameliorate soils polluted by metals.

[Therapeutic use of robotics in children with Autism Spectrum Disorder]. / Uso terapéutico de robótica en niños con Trastorno del Espectro Autista.

INTRODUCTION: Robot-assisted Therapy (RAT) can improve the behavior of children with Autism Spectrum Disorder (ASD) in a spontaneous and entertaining way. There are no previous experiences of this type of inter vention in our country. OBJECTIVE: To describe a clinical experience of using RAT and its impact on the behaviors of a group of children with ASD, in a therapeutic context. PATIENTS AND METHOD: Quasi experimental clinical experience type study. 4 children with a clinical diagnosis of ASD were selected, supported by the ADOS-2 (Autism Diagnostic Observation Schedule); aged between 9 and 13 years, and normal IQ according to the WISC-III (Wechsler Intelligence Scale for Children). This study was approved by the Central Metropolitan Ethics Committee. Patients attended 10 structured robot-as sisted therapy sessions, working collaboratively in pairs. Workshop attendance and parent and child satisfaction were evaluated through surveys, the adaptive behavior with the Vineland scale, and so cial interaction with video coding guidelines. RESULTS: Patients presented a very good adherence and satisfaction with the activity. There was an improvement in socialization behaviors and social age. Video-coding showed an increase in social interaction and improvement in the behavior of the pa tients after attending workshops. CONCLUSIONS: We observed that the experience with RAT, adapted to the context of a Chilean public health center, was highly attractive and beneficial for patients with ASD, improving core symptoms such as difficulties in social interaction and behavioral problems.

[Children and adolescents with intellectual disabilities studied with genetic tests according to their clinical phenotype]. / Estudio genético de acuerdo a características fenotípicas de niños y adolescentes con discapacidad intelectual de etiología indeterminada.

INTRODUCTION: Intellectual disability (ID) is a neurodevelopmental disorder characterized by limitations in intellec tual and adaptive functioning, of various etiologies, including genetic causes. OBJECTIVE: to describe genetic studies carried out in a series of children and adolescents with ID of previously undetermined etiology, considering their phenotypic characteristics. PATIENTS AND METHOD: Descriptive study of a series of patients with ID aged 6 to 18 years. Clinical records, cognitive assessment results (Wechsler -TADI), and genetic study performed were reviewed. They were classified according to phenotypic characteristics into Group 1 patients without a specific phenotype, Group 2: patients with Angel- man- and Rett-like neurodevelopmental disorders phenotype, and Group 3: patients with difficult- to-control seizures. Group 1 was studied with CMA and Groups 2 and 3 with specific genetic panels. RESULTS: 18 patients were described, average age 11 years, male predominance, non-consanguineous parents, and with history of psychomotor retardation. Common comorbidities were epilepsy, autism spectrum disorder (ASD), and behavioral difficulties. Most had a neurological examination without focus and had TADI with very poor developmental ages. In Group 1, there was one patient with a 16p11.2 microdeletion and in Group 3 a duplication of the IQSEC2 gene was found in a patient with difficult-to-control seizures. CONCLUSIONS: The phenotypic characteristics allow to guide the choice of specific genetic studies in children and adolescents with ID of previously undetermined etiology to approach the etiological diagnosis.

Autism screening and conditional cash transfers in Chile: Using the Quantitative Checklist (Q-CHAT) for early autism detection in a low resource setting.

LAY ABSTRACT: Getting a diagnosis of autism can take long, because autism is different across people, but also because it depends on the way it gets diagnosed. This is especially important in poorer countries or in the case of poor people living in wealthier countries that have significant groups of disadvantaged communities. We adapted a 10-item version of the Q-CHAT-25 questionnaire for use in routine health check-ups programme in Chile and recruited 287 participants under the age of three divided into three groups: Controls (125), Developmental Delay (149) and Autism Spectrum Condition (13). Our results show that a short questionnaire for autism screening can be successfully applied in a health-check programme in poor resource settings. Our results show that our questionnaire had good overall performance, not different to its longer version, the Q-CHAT-25. Our questionnaire was autism specific, with good sensitivity and reliability, and is suitable to be used in a screening setting. This study provides evidence that the implementation of Autism Spectrum Condition screening programmes using the Q-CHAT-10 provides value for money and improves diagnosis of Autism Spectrum Condition in those participating in routine health check-up programmes in developing countries or poor areas of wealthy countries.

Characterization of Bacteriophages against Pseudomonas Syringae pv. Actinidiae with Potential Use as Natural Antimicrobials in Kiwifruit Plants.

Pseudomonas syringae pv. actinidiae (Psa) is the causal agent of a bacterial canker in kiwifruit plants and has caused economic losses worldwide. Currently, the primary strategies to control this pathogen include the use of copper-based compounds and even antibiotics. However, the emergence of isolates of Psa that are resistant to these agrochemicals has raised the need for new alternatives to control this pathogen. Bacteriophages have been proposed as an alternative to control bacterial infections in agriculture, including Psa. Here, we show the isolation and characterization of 13 phages with the potential to control Psa infections in kiwifruit plants. The phages were characterized according to their host range and restriction fragment length polymorphism (RFLP) pattern. Four phages were selected according to their lytic effect on the bacteria and their tolerance to different environmental conditions of pH (4-7), temperature (4-37 °C), and solar radiation exposure (30 and 60 min). The selected phages (CHF1, CHF7, CHF19, and CHF21) were sequenced, revealing a high identity with the podophage of Psa phiPSA2. In vitro assays with kiwifruit leaf samples demonstrated that the mixture of phages reduced the Psa bacterial load within three hours post-application and was able to reduce the damage index in 50% of cases. Similarly, assays with kiwifruit plants maintained in greenhouse conditions showed that these phages were able to reduce the Psa bacterial load in more than 50% of cases and produced a significant decrease in the damage index of treated plants after 30 days. Finally, none of the selected phages were able to infect the other bacteria present in the natural microbiota of kiwifruit plants. These results show that bacteriophages are an attractive alternative to control Psa infections in kiwifruit plants.

Chilean regulations on metal-polluted soils: The need to advance from adapting foreign laws towards developing sovereign legislation.

Chile as a major international Cu producer faces serious soil contamination issues in mining areas. Currently Chile does not have any specific law governing the maximum permissible concentrations of metals in soils to protect ecosystems and human health. Chile heavily relies on the use of environmental laws of 14 foreign countries; the choice of the country depends on the similarity of its environmental conditions with those in Chile. In this study, we used an online database to compare the similarity of Chilean rocks to those in foreign countries. Likewise, we performed soil sampling and determined the background concentrations of Cu, As, Pb, and Zn in soils of the Aconcagua basin, the largest river basin in the Valparaiso Region of central Chile. The results showed that geochemical patterns in Chile have the greatest resemblance to New Zealand, Mexico, and Italy. The background Cu concentration in the Aconcagua basin (134 mg kg-1) exceeded the legislated limits of New Zealand (100 mg kg-1) and Italy (120 mg kg-1), whereas the background Zn concentration (200 mg kg-1) exceeded the legislated limit of Italy (150 mg kg-1). Due to the elevated natural abundance of Cu and Zn in Chile, international laws should not be applied in Chile for the assessment of soil contamination. In addition, we assessed ecological risk using the results of our previous studies obtained by analyzing native field-contaminated soils of the Valparaiso region. In the Aconcagua basin, Cu posed high risk for plants in 11% of the samples, whereas As posed high risk for earthworms in 48% of the samples. We suggest that future studies are required to search for other organisms that can serve as biomarkers of metal toxicity because our previous studies were limited to plants and earthworms. Importantly, As posed high risk to human health in 25% of the samples in our study. There is a need for future studies to demonstrate empirically an association between soil As and children's blood As in order to establish the national threshold values of soil As to protect human health. We conclude that there is an urgent need in Chile to advance from the current approach of adapting foreign laws to developing Chilean sovereign environmental legislation.

Insights Into the Microbiology of the Chaotropic Brines of Salar de Atacama, Chile.

Microbial life inhabiting hypersaline environments belong to a limited group of extremophile or extremotolerant taxa. Natural or artificial hypersaline environments are not limited to high concentrations of NaCl, and under such conditions, specific adaptation mechanisms are necessary to permit microbial survival and growth. Argentina, Bolivia, and Chile include three large salars (salt flats) which globally, represent the largest lithium reserves, and are commonly referred to as the Lithium Triangle Zone. To date, a large amount of information has been generated regarding chemical, geological, meteorological and economical perspectives of these salars. However, there is a remarkable lack of information regarding the biology of these unique environments. Here, we report the presence of two bacterial strains (isolates LIBR002 and LIBR003) from one of the most hypersaline lithium-dominated man-made environments (total salinity 556 g/L; 11.7 M LiCl) reported to date. Both isolates were classified to the Bacillus genera, but displayed differences in 16S rRNA gene and fatty acid profiles. Our results also revealed that the isolates are lithium-tolerant and that they are phylogenetically differentiated from those Bacillus associated with high NaCl concentration environments, and form a new clade from the Lithium Triangle Zone. To determine osmoadaptation strategies in these microorganisms, both isolates were characterized using morphological, metabolic and physiological attributes. We suggest that our characterization of bacterial isolates from a highly lithium-enriched environment has revealed that even at such extreme salinities with high concentrations of chaotropic solutes, scope for microbial life exists. These conditions have previously been considered to limit the development of life, and our work extends the window of life beyond high concentrations of MgCl2, as previously reported, to LiCl. Our results can be used to further the understanding of salt tolerance, most especially for LiCl-dominated brines, and likely have value as models for the understanding of putative extra-terrestrial (e.g., Martian) life.

Genetic and Phenotypic Characterization of Indole-Producing Isolates of Pseudomonas syringae pv. actinidiae Obtained From Chilean Kiwifruit Orchards.

In recent years, Chilean kiwifruit production has been affected by the phytopathogen Pseudomonas syringae pv. actinidiae (Psa), which has caused losses to the industry. In this study, we report the genotypic and phenotypic characterization of 18 Psa isolates obtained from Chilean kiwifruits orchards between 2012 and 2016 from different geographic origins. Genetic analysis by multilocus sequence analysis (MLSA) using four housekeeping genes (gyrB, rpoD, gltA, and gapA) and the identification of type III effector genes suggest that the Chilean Psa isolates belong to the Psa Biovar 3 cluster. All of the isolates were highly homogenous in regard to their phenotypic characteristics. None of the isolates were able to form biofilms over solid plastic surfaces. However, all of the isolates formed cellular aggregates in the air-liquid interface. All of the isolates, except for Psa 889, demonstrated swimming motility, while only isolate Psa 510 demonstrated swarming motility. The biochemical profiles of the isolates revealed differences in 22% of the tests in at least one Psa isolate when analyzed with the BIOLOG system. Interestingly, all of the isolates were able to produce indole using a tryptophan-dependent pathway. PCR analysis revealed the presence of the genes aldA/aldB and iaaL/matE, which are associated with the production of indole-3-acetic acid (IAA) and indole-3-acetyl-3-L-lysine (IAA-Lys), respectively, in P. syringae. In addition, IAA was detected in the cell free supernatant of a representative Chilean Psa strain. This work represents the most extensive analysis in terms of the time and geographic origin of Chilean Psa isolates. To our knowledge, this is the first report of Psa being able to produce IAA. Further studies are needed to determine the potential role of IAA in the virulence of Psa during kiwifruit infections and whether this feature is observed in other Psa biovars.

Zinc alleviates copper toxicity to symbiotic nitrogen fixation in agricultural soil affected by copper mining in central Chile.

According to the Terrestrial Biotic Ligand Model, other cations might compete with Cu+2 for biotic ligand sites and provide a protective effect. In particular, evidence suggests Zn may alleviative Cu toxicity. No study, to the best of our knowledge, has focused explicitly on the alleviating effect Zn might have on Cu toxicity to soil microorganisms in field-contaminated soils. The aim of this study was to investigate the alleviating effect Zn might have on Cu toxicity to symbiotic nitrogen fixation in agricultural soils affected by copper mining in central Chile. The bioassay estimated the symbiotic nitrogen fixation capacity of a population of rhizobia in a specified soil, using the soil as inocula for Phaseolus vulgaris L. grown in a soil-less system (pots with perlite) irrigated with a sterile nitrogen-free nutrient solution. Among all soil physicochemical characteristics, the Cu/Zn ratio best explained changes in symbiotic nitrogen fixation. The effective concentration 50% (EC50) of Cu/Zn ratio for symbiotic nitrogen was equal to 1.2, with 95% confidence interval of 1.0-1.3. Symbiotic nitrogen fixation decreased with increased Cu/Zn ratio, thus suggesting that Zn alleviates Cu toxicity to nitrogen fixing microorganisms.