Actividad antibacteriana de actinomicetos frente a Escherichia coli y Staphylococcus aureus multirresistentes de origen hospitalario / Antibacterial activity of actimycetes against multiresistant Escherichia coli and Staphylococcus aureus isolated from a hospital

Introducción: La resistencia a los antimicrobianos (RAM) es un problema de salud pública que manifiesta la disminuida eficacia de estos agentes en la prevención y tratamiento de una proporción cada vez más amplia de patologías. Los actinomicetos son un grupo bacteriano importante de productores de metabolitos activos contra patógenos. Objetivo: Aislar actinomicetos del bosque tropical de Nariño, con potencial producción de metabolitos inhibitorios contra bacterias multidrogo-resistentes. Materiales y métodos: Se tomaron muestras de suelo de Bosque Tropical Húmedo de la Reserva Natural del Río Ñambí, se analizaron microbiológica y molecularmente. Se estimuló la producción in vitro de metabolitos secundarios y evaluó el efecto inhibitorio de estos extractos contra las bacterias multidrogo-resistentes Escherichia coli y Staphylococcus aureus. Resultados: Se obtuvieron 11 aislados presuntivos, se confirmó que cuatro de ellos correspondieron al género Streptomyces sp. Las pruebas de inhibición contra bacterias multidrogo-resistentes E. coli y S. aureus, permitieron verificar que el aislado P3772 fue el más eficiente en la inhibición de los patógenos. Conclusiones: Todos los actinomicetos evaluados presentan actividad antibacteriana contra al menos una de las bacterias patógenas estudiadas; destacando el aislado P3772, que inhibe a E. coli y S. aureus. Se espera caracterizar los compuestos vinculados a la actividad antibacteriana.

Introduction: Antimicrobial resistance (AR) is a public health problem that reveals the diminished efficacy of these agents in the prevention and treatment of an increasingly larger number of pathologies. Actinomycetes are an important bacterial producer group of metabolites that are active against pathogens. Objective: To isolate actinomycetes from the tropical forest of Nariño (Colombia), which have the potential to produce inhibitory metabolites against multi-drug resistant bacteria. Materials and methods: Soil samples were taken from the Humid Tropical Forest of the Río Ńambí Natural Reserve and analyzed through microbiological and molecular assays. In vitro production of secondary metabolites was first stimulated, followed by the assessment of the inhibitory effect of these extracts against multi-drug resistant Escherichia coli and Staphylococcus aureus. Results: 11 presumptive isolates were obtained, confirming that four of them corresponded to the Streptomyces sp. genus. The bacterial isolate P3772 was identified as the one with the highest inhibitory effect against multi-drug resistant E. coli and S. aureus. Conclusions: All the actinomycetes evaluated presented antibacterial activity. The isolate P3772 stands out, which inhibited both E. coli and S. aureus. The compounds associated with this antibacterial activity will be characterized in future studies.

Polyhydroxyalkanoate biosynthesis by oxalotrophic bacteria from high Andean soil / Biosíntesis de polihidrxialcanoato por bacterias oxalotróficas de suelos altoandinos / Biossínteses de polihidroxialcanoato por bactérias oxalotróficas de solos andinos

Abstract Oxalate is a highly oxidized organic acid anion used as a carbon and energy source by oxalotrophic bacteria. Oxalogenic plants convert atmospheric CO2 into oxalic acid and oxalic salts. Oxalate-salt formation acts as a carbon sink in terrestrial ecosystems via the oxalate-carbonate pathway (OCP). Oxalotrophic bacteria might be implicated in other carbon-storage processes, including the synthesis of polyhydroxyalkanoates (PHAs). More recently, a variety of bacteria from the Andean region of Colombia in Narino have been reported for their PHA-producing abilities. These species can degrade oxalate and participate in the oxalate-carbonate pathway. The aim of this study was to isolate and characterize oxalotrophic bacteria with the capacity to accumulate PHA biopolymers. Plants of the genus Oxalis were collected and bacteria were isolated from the soil adhering to the roots. The isolated bacterial strains were characterized using biochemical and molecular biological methods. The consumption of oxalate in culture was quantified, and PHA production was monitored in batch fermentation. The polymeric composition was characterized using gas chromatography. Finally, a biosynthetic pathway based on our findings and on those from published sources is proposed. Strains of Bacillus spp. and Serratia sp. were found to metabolize calcium oxalate and synthesize PHA.

Resumen El oxalato es un anión de ácido orgánico altamente oxidado usado como fuente carbono y energía por bacterias oxalotróficas. Las plantas oxalogénicas convierten CO2 atmosférico en ácido oxálico y sales oxálicas. La formación de sales de oxalato actúa como un sumidero de carbono en ecosistemas terrestres via oxalato-carbonato (OCP). Las bacterias oxalotróficas podrían estar implicadas en otros procesos de almacenamiento de carbono, incluyendo la síntesis de polihidroxialcanoatos (PHAs). Recientemente, una variedad de bacterias de la región andina colombiana en Nariño ha sido reportada por su habilidad para producir PHAs. Estas especies pueden degradar oxalato y participar en la vía del oxalato-carbonato. El objetivo de este estudio fue aislar y caracterizar bacterias oxalotróficas con capacidad de acumular biopolímeros PHA. Se colectaron plantas del genero Oxalis y se aislaron bacterias del suelo adheridas a las raíces. Las cepas bacterianas aisladas se caracterizaron usando métodos bioquímicos y de biología molecular. Se cuantificó el consumo de oxalato en cultivo, y se monitoreó la producción de PHA en fermentación por lotes. La composición polimérica se caracterizó usando cromatografía de gases. Finalmente, se propone una via biosintética basada en nuestros hallazgos y en los de otras fuentes publicadas. Se encontró que las cepas de Bacillus spp. y Serratia sp. metabolizan oxalato de calcio y sintetizan PHA.

Resumo O oxalato é um ânion de ácido orgânico altamente oxidado utilizado como fonte de carbono e nergía por bactérias oxalotróficas. As plantas oxalogênicas convertem CO2 atmosférico em ácido oxálico e sais oxálicos. A formação de sais de oxalato atua como um sumidouro de carbono em ecossistemas terrestres via oxalato-carbono (OCP). As bactérias oxalotróficas poderiam estar envolvidas em outros processos de armazenamento de carbono, incluindo a sínteses de polihidroxialcanoatos (PHAs). Recentemente, uma variedade de bactérias da região Andina colombiana no Departamento de Nariño foi reportada devido a sua habilidade para produzir PHAs. Estas espécies podem degradar oxalato e participar na via oxalato-carbono. O objetivo de esse estudo foi isolar e caracterizar bactérias oxalotróficas com capacidade de acumular biopolímeros PHA. Plantas do género Oxalis foram coletadas e se isolaram bactérias do solo aderido a suas raízes. As cepas bacterianas isoladas se caracterizaram utilizando métodos bioquímicos e de biologia molecular. O consumo de oxalato em cultivo foi quantificado, e a produção de PHA foi monitorada em fermentação por lotes. A composição polimérica se caracterizou utilizando Cromatografia de Gases. Finalmente, se propõe uma via biossintética baseada em nossos resultados juntamente com resultados da literatura. Se encontrou que as cepas de Bacillus spp. e Serratia sp. metabolizam oxalato de cálcio e sintetizam PHA.

Nontuberculous mycobacteria in milk from positive cows in the intradermal comparative cervical tuberculin test: implications for human tuberculosis infections.

Although the tuberculin test represents the main in vivo diagnostic method used in the control and eradication of bovine tuberculosis, few studies have focused on the identification of mycobacteria in the milk from cows positive to the tuberculin test. The aim of this study was to identify Mycobacterium species in milk samples from cows positive to the comparative intradermal test. Milk samples from 142 cows positive to the comparative intradermal test carried out in 4,766 animals were aseptically collected, cultivated on Lowenstein-Jensen and Stonebrink media and incubated for up to 90 days. Colonies compatible with mycobacteria were stained by Ziehl-Neelsen to detect acid-fast bacilli, while to confirm the Mycobacterium genus, conventional PCR was performed. Fourteen mycobacterial strains were isolated from 12 cows (8.4%). The hsp65 gene sequencing identified M. engbaekii (n=5), M. arupense (n=4), M. nonchromogenicum (n=3), and M. heraklionense (n=2) species belong to the Mycobacterium terrae complex. Despite the absence of M. tuberculosis complex species in the milk samples, identification of these mycobacteria highlights the risk of pathogen transmission from bovines to humans throughout milk or dairy products, since many of mycobacterial species described here have been reported in pulmonary and extrapulmonary diseases both in immunocompetent and immunocompromised people.

Mathematical model for the growth of Mycobacterium tuberculosis in the granuloma.

In this work we formulate a model for the population dynamics of Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB). Our main interest is to assess the impact of the competition among bacteria on the infection prevalence. For this end, we assume that Mtb population has two types of growth. The first one is due to bacteria produced in the interior of each infected macrophage, and it is assumed that is proportional to the number of infected macrophages. The second one is of logistic type due to the competition among free bacteria released by the same infected macrophages. The qualitative analysis and numerical results suggests the existence of forward, backward and S-shaped bifurcations when the associated reproduction number R0 of the Mtb is less unity. In addition, qualitative analysis of the model shows that there may be up to three bacteria-present equilibria, two locally asymptotically stable, and one unstable.

Screening of polyketide genes from Brazilian Atlantic Forest soil / Evaluación de genes biosintéticos de policétidos provenientes de suelo de la selva atlántica brasileñae / Avaliação de genes biossintéticos de policetídeos provenientes do solo da Mata Atlântica Brasileira

Abstract Soil is a large source of microorganisms with potential to produce bioactive compounds. Since most of them cannot be cultured, metagenomics has become a useful tool in order to evaluate this potential. The aim of this study was to screen biosynthetic polyketide genes (PKS) present in a metagenomic library constructed from a soil sample isolated from the Brazilian Atlantic Forest. The library comprises 5000 clones with DNA inserts between 40 and 50 Kb. The characterization of the biosynthetic gene clusters of these molecules is a promising alternative to elucidate the biotechnological potential of bioactive compounds in microbial communities. The PKS genes were screened using degenerated primers. The positive clones for PKS systems were isolated, and their nucleotide sequences analysed with bioinformatics tools. The screening yielded two positive clones for PKS II genes. Furthermore, variations in the sequences of the PKS II genes from the metagenomic library were observed when compared with sequences of ketosynthases' databases. With these findings we gain insight into the possible relation between new biosynthetic genes and the production of new secondary metabolites.

Resumen El suelo es una fuente importante de microrganismos con potencial para producir compuestos bioactivos. Dado que la gran mayoría de estos microorganismos no puede cultivarse, la metagenómica se ha convertido en una herramienta útil para evaluar dicho potencial. El objetivo del presente estudio fue evaluar los genes biosintéticos de policétidos (PKS) presentes en una biblioteca metagenómica construida a partir de una muestra de suelo aislada de la selva atlántica brasileña. La biblioteca comprende 5000 clones con insertos de DNA entre 40 y 50 Kb. La caracterización de clústeres de genes biosintéticos de estas moléculas es una alternativa promisoria para elucidar el potencial biotecnológico de los compuestos bioactivos en comunidades microbianas. Los genes biosintéticos de PKS se evaluaron usando cebadores degenerados. Se aislaron los clones positivos para sistemas PKS y sus secuencias de nucleótidos se analizaron con herramientas bioinformáticas. La evaluación arrojó dos clones positivos para genes de PKS II. Además, se observaron variaciones en las secuencias de genes de PKS II de la biblioteca metagenómica cuando se compararon con las secuencias de las bases de datos de cetosintasas. Estos hallazgos proporcionan nueva información sobre la posible relación entre nuevos genes biosintéticos y la producción de nuevos metabolitos secundarios.

Resumo O solo é uma fonte de importante de microrganismos com potencial para produzir compostos bioativos. Considerando que a maioria destes microrganismos não se pode cultivar, a metagenômica tem se convertido em uma ferramenta útil para avaliar este potencial. O objetivo deste estudo foi avaliar os genes biossintéticos de policetídeos (PKS) presentes em uma biblioteca metagenômica construída a partir de uma amostra de solo isolada da Mata Atlântica brasileira. A biblioteca compreende 5000 clones com insertos de DNA entre 40 e 50 Kb. A caracterização de clusters de genes biossintéticos destas moléculas é uma alternativa promissora para elucidar o potencial biotecnológico de compostos bioativos em comunidades microbianas. Os genes PKS foram avaliados usando primers degenerados. Os clones positivos para sistemas PKS foram isolados e suas sequências de nucleotídeos foram analisadas com ferramentas de bioinformática. A avaliação forneceu dois clones positivos para genes PKS II. Além disso, variações nas sequências dos genes PKS II da biblioteca metagenômica foram observadas quando comparadas com sequências da base de dados de cetosintases. Com estas descobertas obtivemos uma visão sobre uma possível relação entre novos genes biossintéticos e a produção de novos metabólitos secundários.

Eficiencia en la reducción de Cromo por una bacteria silvestre en un tratamiento tipo Batch utilizando como sustrato agua residual del municipio de Pasto, Colombia / Efficiency in the reduction of chromium by a wild bacterium in a Batch treatment type using residual water substrate from the municipality of Pasto, Colombia

Resumen Objetivo: Evaluar la eficiencia en la reducción de Cromo en un tratamiento tipo Batch, utilizando como sustrato agua residual municipal inoculada con una bacteria silvestre. Materiales y métodos: Se verificó a escala de laboratorio el porcentaje de reducción de Cromo hexavalente de tres bacterias silvestres previamente aisladas de agua residual del Río Pasto (Bacillus thuringiensis, Bacillus amyloliquefaciens y Paenibacillus sp.); se seleccionó el aislado que presentó mayor porcentaje de reducción de Cr y fue sometido a diferentes tratamientos. El análisis de los resultados se hizo mediante estadística descriptiva. Resultados: B. thuringiensis, B. amyloliquefaciens, y Paenibacillus sp., presentaron porcentajes de reducción de Cr (VI) de 82,01%; 80,85% y 79,27%, respectivamente. Se determinó que el tercer tratamiento (agua sin esterilizar del Río Pasto con B. thuringiensis) presentó diferencias significativas respecto a los demás (p = 0,0001 α = 0,05), concluyendo que B. thuringiensis reduce en mayor proporción el Cr (VI), los resultados encontrados en esta investigación son promisorios en el campo de la biorremediación de efluentes contaminados con Cromo ya que pueden ser tomados como base para implementar estrategias de biorremediación a gran escala. Conclusión: La bacteria B. thuringiensis presentó alta eficiencia en la reducción de Cromo hexavalente (99,42%), cuando fue implementada en un tratamiento a escala de laboratorio de agua residual sin esterilizar.

Abstract Objective: To evaluate the efficiency in the reduction of chromium in a Batch treatment type, using municipal residual water substrate inoculated with a wild bacterium. Materials and methods: The reduction percentage of hexavalent chromium of three wild bacteria previously isolated from residual water from the Pasto River was verified at laboratory scale (Bacillus thuringiensis, Bacillus amyloliquefaciens and Paenibacillus sp.); the isolated that showed the highest percentage of reduction of Cr was selected and was subjected to different treatments. The analysis of results was done using descriptive statistics. Results: B. thuringiensis, B. amyloliquefaciens, and Paenibacillus sp., presented percentages of reduction of Cr (VI) of 82,01%; 80,85% and 79,27%, respectively. It was determined that the third treatment (nonsterile water from the Pasto River with B. thuringiensis) presented significant differences with regard to the other (p = 0.0001 α = 0.05), concluding that B. thuringiensis reduces in greater proportion the Cr (VI). The results found in this research are promising in the field of bioremediation of contaminated effluents with Chrome since they may be taken as the basis for implementing strategies of bioremediation on a large scale. Conclusion: The bacteria B. thuringiensis presented high efficiency in the reduction of hexavalent chromium (99.42%) when implemented in a treatment at laboratory scale of residual nonsterile water.

Degradación de oxalato por bacterias oxalotróficas asociadas a plantas del género Oxalis sp en regiones Andinas del departamento de Nariño, Colombia / Oxalate degradation by oxalotrophic bacteria associated with plants of the genus Oxalis sp in the Andean region of the department of Nariño, Colombia

Introducción: El ácido oxálico (H2C2O4) y las sales de oxalato son sustancias altamente oxidadas y consideradas tóxicas para algunos sistemas biológicos, incluido el humano, no obstante, pueden ser utilizadas como fuente de carbono y energía por algunas comunidades bacterianas, denominadas oxalotróficas, las cuales por su capacidad metabólica forman parte de la ruta biogeoquímica oxalato-carbonato (OCP, oxalate-carbonate pathway). Objetivo: Aislar y caracterizar bacterias oxalotróficas a partir de plantas del género Oxalis sp., de zonas alto-andina de Nariño- Colombia. Materiales y métodos: Se recolectaron muestras de suelo rizosférico de plantas oxalogénicas que fueron analizadas con parámetros fisicoquímicos y se utilizó un medio selectivo Schlegel para el aislamiento de bacterias oxalotróficas. Resultados: Las bacterias aisladas en medio selectivo Schlegel fueron caracterizadas bioquímicamente como: (Serratia fonticola, Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus vallismortis y Bacillus cereus). Estas especies fueron capaces de degradar oxalato e incrementar el pH producto de la degradación. Conclusión: Este tipo de bacterias pueden ser estudiadas en trabajos complementarios para evaluar su potencial como biofertilizantes y/o alternativas de bioremediación en suelos ácidos. El estudio a pesar de ser indicativo a nivel biológico, puede en un futuro y con base en mayores soportes en investigación, tornarse en una promisoria aplicación para reducir el oxalato de calcio en los alimentos de consumo diario que presentan un renglón de importancia agrícola en la región, potencialmente dañinos para la función renal.

Introduction: Oxalic acid (H2C2O4) and oxalate salts are highly oxidized substances, which are considered as toxic for some biological systems, including the human being; however, they can be used as a source of carbon and energy for some bacterial communities called oxalotrophic which are part of the so called oxalate-carbonate geochemistry pathway (OCP) due to its metabolic capacity. Objective: The aim of this research was to isolate and characterize oxalotrophic bacteria from plants of the genus Oxalis sp. in the high-andean zone of the department of Nariño, Colombia. Materials and methods: Samples of rhizosphere soil from oxalogenic plants were analyzed with physicochemical parameters and a Schlegel selective medium was used to isolate oxalotrophic bacteria. Results: The isolated bacteria through Schlegel selective medium were identified biochemically as: (Serratia fonticola, Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus vallismortis and Bacillus cereus). The identified species play an important role in the rhizosphere soil, principally for the capacity to increase the pH during oxalate consumption. Conclusion: This type of bacteria can be studied in additional studies to evaluate their potential as bio-fertilizers and/or bioremediation alternatives in acid soils. Despite that the study is indicative at a biological level, it can become a promising application, in the future and with greater support in research, to reduce the calcium oxalate in food of daily consumption which represent an agricultural important line in the region and are potentially harmful to kidney finction.

Mathematical modeling on bacterial resistance to multiple antibiotics caused by spontaneous mutations.

We formulate a mathematical model that describes the population dynamics of bacteria exposed to multiple antibiotics simultaneously, assuming that acquisition of resistance is through mutations due to antibiotic exposure. Qualitative analysis reveals the existence of a free-bacteria equilibrium, resistant-bacteria equilibrium and an endemic equilibrium where both bacteria coexist.