Identificacción de infecciones de vías urinarias, agente etiológico, sensibilidad a los antimicrobianos y factores asociados en los habitantes de la Comunidad de San Francisco de Cuchocorral. El Tambo-Cañar 2015

ANTECEDENTES.- Las infecciones urinarias representan un grave problema de salud que afecta a la población en general. En El Tambo representan una de las principales causas de consulta externa y hospitalización. OBJETIVO.- Identificar la prevalencia de infecciones de vías urinarias, agente etiológico, sensibilidad a los antimicrobianos y factores asociados en los habitantes de la comunidad de San Francisco de Cuchocorral, El Tambo 2015. METODOLOGÍA.- El estudio fue descriptivo de corte transversal. El universo fue de 208 habitantes y la muestra de 160 personas de la comunidad. Se realizó el examen elemental y microscópico de orina, el urocultivo y el antibiograma. Previo a la recolección de las muestras, los participantes firmaron el consentimiento o asentimiento informado y llenaron una encuesta. Se analizaron las muestras en el laboratorio de Microbiología de la Facultad de Ciencias Médicas. Para relacionar los resultados obtenidos y las variables de estudio se utilizó el programa SPSS V22 y Excel para la estadística descriptiva. RESULTADOS.- La prevalencia de infección urinaria fue del 10%, siendo el 100% de personas con infección del sexo femenino y de ellas el 44% mujeres de 19 – 45 años. Escherichia coli fue el microorganismo más frecuente con el 87,5% y mostró resistencia para Trimetoprim sulfametoxazol y Gentamicina en un 28,6%. Proteus y Klebsiella presentaron resistencia para Ampicilina sulbactam y Fosfomicina en un 100%. CONCLUSIÓN.- Contribuimos con datos epidemiológicos sobre prevalencia de infecciones de vías urinarias, agente etiológico y sensibilidad a los antimicrobianos en los habitantes de la comunidad de San Francisco de Cuchocorral

Degradación bacteriana de cianuro y compuestos nitrogenados tóxicos

La ruta de asimilación de cianuro en P. pseudoalcaligenes CECT5344 transcurre a través de un nitrilo formado por la reacción química del cianuro con el oxalacetato, siendo este último acumulado como consecuencia de la acción conjunta de una malato:quinona oxidoreductasa (MQO) y la oxidasa terminal resistente a cianuro (CioAB) (Luque-Almagro et al., 2011b). Los nitrilos pueden ser convertidos en amonio por la acción de una nitrilasa o un sistema nitrilo hidratasa/amidasa. Con el objetivo de elucidar la ruta de asimilación de cianuro en P. pseudoalcalígenes CECT5344, se ha analizado el proteoma de este microorganismo en condiciones cianotróficas frente a nitrato como fuente de nitrógeno como control. En este estudio se identificaron proteínas relacionadas con la ruta de asimilación de cianuro en la estirpe CECT5344, que aparecían inducidas por cianuro, como NitB y NitG, cuyos genes se encuentran localizados en la agrupación génica nit1C. Además de NitB y NitG, de función desconocida, la agrupación génica nit1C codifica un regulador transcripcional del tipo Fis dependiente de σ54 (NitA), una nitrilasa (NitC), una proteína que pertenece a la superfamilia S-adenosilmetionina (NitD), un miembro de la superfamilia N-aciltransferasa (NitE), un polipéptido de la familia AIRS/GARS (NitF) y una oxidorreductasa dependiente de NADH (NitH). Un análisis transcripcional mediante RT-PCR determinó que los genes nitBCDEFGH se cotranscriben, mientras que el gen regulador nitA se transcribe de forma divergente. Además, resultados obtenidos por RT-PCR confirman que la expresión de los genes nitBCDEFGH está inducida por cianuro y reprimida por amonio. La relación entre el cianuro y el grupo de genes nit1C queda patente por el fenotipo de los mutantes deficientes nitA, nitB y nitC, incapaces de usar complejos cianuro-metálicos o 2-hidroxinitrilos como única fuente de nitrógeno. Todos estos datos indican que la nitrilasa NitC, junto con la proteína NitB, utilizan de forma específica determinados nitrilos alifáticos como sustrato, entre los que se encuentran el formado durante la asimilación de cianuro (Estepa et al., 2012). Además, entre las proteínas inducidas por cianuro se identificaron una dihidropicolinato sintasa (DapA), una fosfoserina transaminasa (SerC) y una proteína de función desconocida (Orf1), las tres codificadas por genes del operón cio, una cianasa (CynS), la proteína S6 de la subunidad ribosomal 30S (RpsF), una superóxido dismutasa (SodB), la ferritina (Dps), una oxidorreductasa (Fpr) y un factor de elongación P (EF-P). Una vez identificadas, estas proteínas se han analizado funcionalmente y se han localizado en el genoma de P. pseudoalcaligenes CECT5344 los genes correspondientes, así como los genes adyacentes. La inducción de estas proteínas en condiciones cianotróficas sugiere que el metabolismo del cianuro incluye, además de la resistencia y asimilación de este tóxico, otros procesos biológicos relacionados con el metabolismo del cianato y de algunos aminoácidos, el estrés oxidativo y la homeostasis de hierro, entre otros. Por otra parte, el conocimiento en profundidad y la interpretación de la secuencia génica de P. pseudoalcaligenes CECT5344, así como el análisis comparativo frente a organismos no cianotrofos ha permitido entender algunos de los mecanismos implicados en la resistencia y asimilación de cianuro, lo que permitiría conducir a la posterior mejora del proceso de biodegradación de cianuro. Además, el estudio del genoma de la estirpe CECT5344 permitirá explorar la capacidad de este organismo para ser utilizado en procesos de biorremediación de residuos cianurados en los que se encuentran metales y otros tóxicos (Luque-Almagro et al., 2013; Wibberg et al., 2014). En este trabajo se muestran y discuten los resultados de la secuenciación del genoma de P. pseudoalcaligenes, así como el estudio del análisis filogenético y evolutivo de la cepa, estableciéndose de esta manera relaciones con otras especies en base a los genomas secuenciados de las mismas, entre las que destaca P. mendocina ymp relacionada con P. pseudoalcaligenes CECT5344. El estudio de las características del genoma de P. pseudoalcaligenes CECT5344 ha sido completado con un análisis comparativo frente a los genomas de otras especies de Pseudomonas, encontrándose así semejanzas y diferencias en cuanto a la distribución génica funcional. Por último, se muestra un análisis del genoma de P. pseudoalcaligenes CECT5344 en relación con los genes implicados probablemente en los procesos de asimilación de cianuro y residuos cianurados, tales como los codificantes de nitrilasas y aquellos implicados en la resistencia a cianuro como los constituyentes del operón cio que codifican la oxidasa terminal insensible a cianuro. Finalmente, se discute la presencia de genes implicados posiblemente en otros procesos con una alto potencial biotecnológico, tales como la producción de bioplásticos y la biodegradación de diversos contaminantes.

Effects of the encapsulation of usnic acid into liposomes and interactions with antituberculous agents against multidrug-resistant tuberculosis clinical isolates

Mycobacterium tuberculosis (Mtb) has acquired resistance and consequently the antibiotic therapeutic options available against this microorganism are limited. In this scenario, the use of usnic acid (UA), a natural compound, encapsulated into liposomes is proposed as a new approach in multidrug-resistant tuberculosis (MDR-TB) therapy. Thus the aim of this study was to evaluate the effect of the encapsulation of UA into liposomes, as well as its combination with antituberculous agents such as rifampicin (RIF) and isoniazid (INH) against MDR-TB clinical isolates. The in vitro antimycobacterial activity of UA-loaded liposomes (UA-Lipo) against MDR-TB was assessed by the microdilution method. The in vitro interaction of UA with antituberculous agents was carried out using checkerboard method. Minimal inhibitory concentration values were 31.25 and 0.98 μg/mL for UA and UA-Lipo, respectively. The results exhibited a synergistic interaction between RIF and UA [fractional inhibitory concentration index (FICI) = 0.31] or UA-Lipo (FICI = 0.28). Regarding INH, the combination of UA or UA-Lipo revealed no marked effect (FICI = 1.30-2.50). The UA-Lipo may be used as a dosage form to improve the antimycobacterial activity of RIF, a first-line drug for the treatment of infections caused by Mtb.

Effect of nitrogen nutrition on yeast ecology and alcoholic fermentation

Mestrado Vinifera Euromaster - Instituto Superior de Agronomia - UL

Avaliação da espécie vegetal Croton cajucara Benth como inibidor de biocorrosão em aço carbono Aisi 1020

Actually in the oil industry biotechnological approaches represent a challenge. In that, attention to metal structures affected by electrochemical corrosive processes, as well as by the interference of microorganisms (biocorrosion) which affect the kinetics of the environment / metal interface. Regarding to economical and environmental impacts reduction let to the use of natural products as an alternative to toxic synthetic inhibitors. This study aims the employment of green chemistry by evaluating the stem bark extracts (EHC, hydroalcoholic extract) and leaves (ECF, chloroform extract) of plant species Croton cajucara Benth as a corrosion inhibitor. In addition the effectiveness of corrosion inhibition of bioactive trans-clerodane dehydrocrotonin (DCTN) isolated from the stem bark of this Croton was also evaluated. For this purpose, carbon steel AISI 1020 was immersed in saline media (3,5 % NaCl) in the presence and absence of a microorganism recovered from a pipeline oil sample. Corrosion inhibition efficiency and its mechanisms were investigated by linear sweep voltammetry and electrochemical impedance. Culture-dependent and molecular biology techniques were used to characterize and identify bacterial species present in oil samples. The tested natural products EHC, ECF and DCTN (DMSO as solvent) in abiotic environment presented respectively, corrosion inhibition efficiencies of 57.6% (500 ppm), 86.1% (500 ppm) and 54.5% (62.5 ppm). Adsorption phenomena showed that EHC best fit Frumkin isotherm and ECF to Temkin isotherm. EHC extract (250 ppm) dissolved in a polar microemulsion system (MES-EHC) showed significant maximum inhibition efficiency (93.8%) fitting Langmuir isotherm. In the presence of the isolated Pseudomonas sp, EHC and ECF were able to form eco-compatible organic films with anti-corrosive properties

Ecogenômica de archaea e monitoramento de comunidades de procariotos redutores de sulfato: aplicações na indústria de petróleo e gás

The human activities responsible for the ambient degradation in the modern world are diverse. The industrial activities are preponderant in the question of the impact consequences for brazilian ecosystems. Amongst the human activities, the petroliferous industry in operation in Potiguar Petroliferous Basin (PPB) displays the constant risk of ambient impacts in the integrant cities, not only for the human populations and the environment, but also it reaches the native microorganisms of Caatinga ground and in the mangrove sediment. Not hindering, the elaboration of strategies of bioremediation for impacted areas pass through the knowledge of microbiota and its relations with the environment. Moreover, in the microorganism groups associated to oil, are emphasized the sulfate-reducing prokaryotes (SRP) that, in its anaerobic metabolism, these organisms participate of the sulfate reduction, discharging H2S, causing ambient risks and causing the corrosion of surfaces, as pipelines and tanks, resulting in damages for the industry. Some ancestries of PRS integrate the Archaea domain, group of microorganisms whose sequenced genomes present predominance of extremophilic adaptations, including surrounding with oil presence. This work has two correlated objectives: i) the detection and monitoring of the gene dsrB, gift in sulfate-reducing prokaryotes, through DGGE analysis in samples of mDNA of a mangrove sediment and semiarid soil, both in the BPP; ii) to relate genomic characteristics to the ecological aspects of Archaea through in silico studies, standing out the importance to the oil and gas industry. The results of the first work suggest that the petrodegraders communities of SRP persist after the contamination with oil in mangrove sediment and in semiarid soil. Comparing the populations of both sites, it reveals that there are variations in the size and composition during one year of experiments. In the second work, functional and structural factors are the probable cause to the pressure in maintenance of the conservation of the sequences in the multiple copies of the 16S rDNA gene. Is verified also the discrepancy established between total content GC and content GC of the same gene. Such results relating ribosomal genes and the ambient factors are important for metagenomic evaluations using PCR-DGGE. The knowledge of microbiota associated to the oil can contribute for a better destination of resources by the petroliferous industry and the development of bioremediation strategies. Likewise, search to lead to the best agreement of the performance of native microbiota in biogeochemical cycles in Potiguar Petroliferous Basin ecosystem

Analysis of the genome content of Lactococcus garvieae by genomic interspecies microarray hybridization

BACKGROUND Lactococcus garvieae is a bacterial pathogen that affects different animal species in addition to humans. Despite the widespread distribution and emerging clinical significance of L. garvieae in both veterinary and human medicine, there is almost a complete lack of knowledge about the genetic content of this microorganism. In the present study, the genomic content of L. garvieae CECT 4531 was analysed using bioinformatics tools and microarray-based comparative genomic hybridization (CGH) experiments. Lactococcus lactis subsp. lactis IL1403 and Streptococcus pneumoniae TIGR4 were used as reference microorganisms. RESULTS The combination and integration of in silico analyses and in vitro CGH experiments, performed in comparison with the reference microorganisms, allowed establishment of an inter-species hybridization framework with a detection threshold based on a sequence similarity of >or= 70%. With this threshold value, 267 genes were identified as having an analogue in L. garvieae, most of which (n = 258) have been documented for the first time in this pathogen. Most of the genes are related to ribosomal, sugar metabolism or energy conversion systems. Some of the identified genes, such as als and mycA, could be involved in the pathogenesis of L. garvieae infections. CONCLUSIONS In this study, we identified 267 genes that were potentially present in L. garvieae CECT 4531. Some of the identified genes could be involved in the pathogenesis of L. garvieae infections. These results provide the first insight into the genome content of L. garvieae.

2013 survey of Iowa groundwater and evaluation of public well vulnerability classifications for contaminants of emerging concern (2015)

Studies in Iowa have long documented the vulnerability of wells with less than 50 feet (15 meters) of confining materials above the source aquifer to contamination from nitrate and various pesticides. Recent studies in Wisconsin have documented the occurrence of viruses in untreated groundwater, even in wells considered to have little vulnerability to contamination from near-surface activities. In addition, sensitive methods have become available for analyses of pharmaceuticals and pesticides. This study represents the first comprehensive examination of contaminants of emerging concern in Iowa’s groundwater conducted to date, and one of the first conducted in the United States. Raw groundwater samples were collected from 66 public supply wells during the spring of 2013, when the state was recovering from drought conditions. Samples were analyzed for 206 chemical and biological parameters; including 20 general water-quality parameters and major ions, 19 metals, 5 nutrients, 10 virus groups, 3 species of pathogenic bacteria, 5 microbial indicators, 108 pharmaceuticals, 35 pesticides and pesticide degradates, and tritium. The wells chosen for this study represent a diverse range of ages, depths, confining material thicknesses, pumping rates, and land use settings. The most commonly detected contaminant group was pesticide compounds, which were present in 41% of the samples. As many as 6 pesticide compounds were found together in a sample, most of which were chloroacetanilide degradates. While none of the measured concentrations of pesticide compounds exceeded current benchmark levels, several of these compounds are listed on the U.S. Environmental Protection Agency’s Contaminant Candidate List and could be subject to drinking water standards in the future. Despite heavy use in the past decade, glyphosate was not detected, and its metabolite, aminomethylphosphonic acid, was only detected in two of 60 wells tested (3%) at the detection limit of 0.02 μg/L. Pharmaceutical compounds were detected in 35% of 63 samples. Of the 14 pharmaceuticals detected, six had reported concentrations above the method reporting limit, with the maximum reported concentration of 826 ng/L for acetaminophen. Diphenhydramine was the only pharmaceutical to have two detections above the reporting limit, at 24.5 and 145 ng/L. Eight pharmaceuticals had confirmed detections at concentrations below the method reporting limit. Caffeine was the most frequently detected pharmaceutical compound (25%), followed by the caffeine metabolite, 1,7-dimethylxanthine (16%). Microorganisms were detected in 21% of the wells using quantitative polymerase chain reaction methodologies. The most frequently detected microorganism was the pepper mild mottle virus (PMMV), a plant pathogen found in human waste. PMMV was detected in 17% of samples at concentrations ranging from 0.4 to 6.38 gene copies per liter. GII norovirus, human polyomavirus, bovine polyomavirus, and Campylobacter were also detected, while adenovirus, enterovirus, GI norovirus, swine hepatitis E, Salmonella, and enterohemmorhagic E. coli were not detected. No correlations were found between viruses or pathogenic bacteria and microbial indicators. Wells with less than 50 feet (15 meters) of confining material were shown to have greater incidence of surface-related contaminants; however, significant relationships (p<0.05) between confining layer thickness and contaminants were only found for nitrate and herbicides.

Use of organic compost containing waste from small ruminants in corn production.

Composting is a useful way of transforming livestock waste into organic fertilizer, which is proven to increase soil nutrient levels, and thus crop yield. Remains from production and slaughter of small ruminants can become a source of important elements for plant growth, such as N, after microorganism-driven decomposition.The aim of this investigation was to evaluate the effects of this compost on soil fertility and on the nutritional status and yield of the corn crop. The experiment was conducted in a Haplic Luvisol in a randomized block design with six treatments and five application rates of the organic compound in Mg ha-1: 3 (half the standard rate), 6 (standard rate), 9 (one and a half times the standard rate), 12 (twice the standard rate), and 24 (four times the standard rate) and an additional treatment with mineral fertilizers (110, 50 e 30 kg ha-1 of N, P2O5 and K2O, respectively), with four blocks. Evaluations were performed for two harvests of rainfed crops, measuring soil fertility, nutritional status, and grain yield. The compost increased P, K, Na and Zn values in the 0.00-0.20 m layer in relation of mineral fertilization in 616, 21, 114 and 90 % with rate 24 Mg ha-1 in second crop. Leaf N, Mg, and S contents, relative chlorophyll content, and the productivity of corn kernels increased in 27, 32, 36, 20 e 85 %, respectively, of low rate (3 Mg ha-1) to high rate (24 Mg ha-1) with of application of the compost. Corn yield was higher with application of organic compost in rate of 24 Mg ha-1 than mineral fertilizer combination in second crop.

Microbiome and resitome and foods of animal origin

To ensure food safety and to prevent food-borne illnesses, rapid and accurate detection of pathogenic agents is essential. It has already been demonstrated that shotgun metagenomic sequencing can be used to detect pathogens and their antibiotic resistance genes in food. In the studies presented in this thesis, the application shotgun metagenomic sequencing has been applied to investigate both the microbiome and resistome of foods of animal origin in order to assess advantages and disadvantages of shotgun metagenomic sequencing in comparison to the cultural methods. In the first study, it has been shown that shotgun metagenomics can be applied to detect microorganisms experimentally spiked in cold-smoked salmon. Nevertheless, a direct correlation between cell concentration of each spiked microorganism and number of corresponding reads cannot be established yet. In the second and third studies, the microbiomes and resistomes characterizing caeca and the corresponding carcasses of the birds reared in the conventional and antibiotic free farms were compared. The results highlighted the need to reduce sources of microbial contamination and antimicrobial resistance not only at the farm level but also at the post-harvest one. In the fourth study, it has been demonstrated that testing a single aliquot of a food homogenate is representative of the whole homogenate because biological replicates displayed overlapping taxonomic and functional composition. All in all, the results obtained confirmed that the application of shotgun metagenomic sequencing represents a powerful tool that can be used in the identification of both spoilage and pathogenic microorganism, and their resistome in foods of animal origin. However, a robust relationship between sequence read abundance and concentration of colony-forming unit must be still established.