Aplicación de PCR a partir de cultivo en la identificación de subespecies del complejo Mycobacterium Tuberculosis / The use of culture-based PCR for the identification of subspecies of the mycobacterium tuberculosis complex

Los métodos diagnósticos clásicos de tuberculosis (TB) se basan en la utilización de baciloscopía y cultivo. La identificación del agente etiológico desde la positivización del cultivo requiere entre 10 y 15 días, mientras que el empleo de la reacción en cadena de la polimerasa (PCR) disminuye el tiempo a 24 h, lo que permite no solo identificar las subespecies del complejo Mycobacterium tuberculosis (CMTB) sino también diferenciarlas de otras especies ambientales clínicamente importantes (MOTT) facilitando el diagnóstico y tratamiento. El objetivo del presente trabajo fue determinar la utilidad de la PCR en la identificación temprana de las micobacterias pertenecientes al CMTB, a partir de cultivos positivos, de pacientes con sospecha de TB, atendidos en un hospital pediátrico de alta complejidad, durante un período de cuatro años. A cada muestra, se le realizó baciloscopía y cultivo en medio líquido. A los cultivos positivos, una inmunocromatografía lateral (TBIDR) y luego PCR. El 4,6% del total de muestras (510/11.162) pertenecientes a 198 pacientes presentó cultivos positivos. Cuatrocientos veintiseis (84%) correspondieron a muestras respiratorias. El rendimiento de la baciloscopía directa fue del 41% (194/470). Cuatrocientos treinta y ocho (86%) resultaron M. tuberculosis, 21 (4%) Mycobacterium bovis, 7 (1%), M. bovis-BCG y 44 (9%) MOTT. La utilización de medios de cultivos líquidos junto con el empleo de PCR favorecen una rápida orientación microbiológica y constituye una estrategia útil para optimizar el manejo clínico de estas infecciones, desde el punto de vista terapéutico y epidemiológico, especialmente en pediatría (AU)

Classical diagnostic methods for tuberculosis (TB) are based on the use of smear microscopy and culture. The identification of the etiological agent from positive culture requires 10 to 15 days, while the use of the polymerase chain reaction (PCR) reduces the time to 24 h, which allows not only to identify the subspecies of the Mycobacterium tuberculosis complex (MTC) but also to differentiate them from clinically important environmental mycobacteria other than tuberculosis (MOTT), facilitating diagnosis and treatment. The aim of this study was to determine the usefulness of PCR in the early identification of mycobacteria belonging to the MTC, from positive cultures of patients with suspected TB seen in a pediatric tertiary hospital over a 4-year period. For each sample, smear microscopy and culture in liquid medium was performed. Positive cultures were subjected to lateral immunochromatography (TBIDR) and then PCR. Of the total number of samples (510/11,162) belonging to 198 patients, 4.6% showed positive cultures; 426 (84%) were respiratory samples. The direct smear microscopy yield was 41% (194/470). Overall, 438 (86%) were found to be M. tuberculosis, 21 (4%) Mycobacterium bovis, 7 (1%), M. bovis-BCG, and 44 (9%) MOTT. The use of liquid culture media together with the use of PCR favors a rapid microbiological orientation and is a useful strategy to optimize the clinical management of these infections, from a therapeutic and epidemiological point of view, especially in children (AU)

Lanthanoid analysis in seawater by seaFAST-SP3™ system in off-line mode and magnetic sector high-resolution inductively coupled plasma source mass spectrometer.

Analysis of lanthanoids in seawater is challenging due to the complex matrix (∼35 g L-1 TDS) and low dissolved concentrations (in ng L-1). A 4-step strict analytical protocol and state-of-the-art technology were implemented and validated in this study. The 4-steps method involves the 1) sample filtration and acidification (pH<2); 2) pre-concentration by the matrix separation system, 3) off-line injection of the eluted sample; and 4) determination of lanthanoids by high-resolution inductively coupled plasma mass spectrometer (HR-ICP-MS). Since there are no certified values for lanthanoids in seawater are available, the method validation was done by analyzing SLEW-3 (estuarine water reference samples) and comparing with other reports and artificial seawater (100 ng L-1 lanthanoid multi-element standard solutions). SLEW-3 recovery varied from 78.6% to 106% and in artificial samples it ranged from 87 to 110%. Low recovery can be explained by complex organic in seawater, because the UV oxidation was not performed in the acidified samples. The variation was ≤10%, except for Gd, Tb, and Yb (11-13.75%). Blanks varied between 0.01 and 0.07 ng L-1, except for La and Ce (0.13-0.21 ng L-1). Blanks represent <5% SLEW-3 values and <1% synthetic seawater. The procedural detection limit varied from 0.01 to 0.03 ng L-1.•Lanthanoids as geochemical tracers in seawaters•A 4-step strict analytical protocol and state-of-the-art technology for lanthanoids analyses in seawaters•Sample pre-concentration system for matrix separation for the detection of ultra-low lanthanoids levels.

Geogenic lanthanoid signature in coastal and marine waters from the southern Gulf of California.

Lanthanoids in the southern Gulf of California (GC) seawater are reported for the first time. Lanthanoids showed differences between peninsular and continental coastline, coastal or marine ecosystems, and dry or rainy season. The chondrite-normalized values showed high variability but followed a same pattern. Light lanthanoids were more enriched than heavy ones. Values of ∑Ln and La/Lu were higher in continental than peninsular coastlines, coastal than adjacent marine ecosystems, and rainy than dry season. Differences were related to the lithology and perturbation degree of the ecosystem watersheds. The chondrite-normalized patterns are typical of geological origin. Slightly negative Ce anomaly was related to the low levels of oxygen in water for the oxidation of Ce (III) to Ce (IV) and its posterior scavenging. Negative δEu anomaly is explained by an influx of fluvial and eolian materials from the upper continental, while a positive Eu anomaly related to hydrothermal vent inputs was non-evidenced.

Dominance patterns in macroalgal and phytoplankton biomass under different nutrient loads in subtropical coastal lagoons of the SE Gulf of California.

Nine macroalgal blooms were studied in five coastal lagoons of the SE Gulf of California. The nutrient loads from point and diffuse sources were estimated in the proximity of the macroalgal blooms. Chlorophyll a and macroalgal biomass were measured during the dry, rainy and cold seasons. Shrimp farms were the main point source of nitrogen and phosphorus loads for the lagoons. High biomasses were found during the dry season for phytoplankton at site 6 (791.7±34.6 mg m(-2)) and during the rainy season for macroalgae at site 4 (296.0±82.4 g m(-2)). Depending on the season, the phytoplankton biomass ranged between 40.0 and 791.7 mg m(-2) and the macroalgal biomass between 1 and 296.0 g m(-2). The bulk biomass (phytoplankton+macroalgal) displayed the same tendency as the nutrient loads entering the coastal lagoons. Phytoplankton and macroalgal biomass presented a significant correlation with the atomic N:P ratio.