Efecto de la exposición a glifosato y ciprofloxacina en bacterias entéricas de renacuajos

La alta carga de agroquímicos y antibióticos en los ambientes acuáticos y los agroecosistemas representa un riesgo para la vida silvestre. Dado que la microbiota intestinal juega un papel fundamental en el funcionamiento de su hospedador y es sensible a una amplia variedad de contaminantes, su estudio permite evaluar la salud de los organismos. En este trabajo estudiamos los efectos de formulaciones comerciales de un herbicida a base de glifosato (GBH) y del antibiótico ciprofloxacina (CIP), por separado y en mezcla, sobre la diversidad de bacterias intestinales de renacuajos del sapo común (Rhinella arenarum). El estudio de la diversidad de bacterias entéricas cultivables de rápido crecimiento y bajo requerimiento nutricional se llevó a cabo utilizando pruebas microbiológicas clásicas e identificación por espectrometría de masas de tiempo de vuelo por desorción/ionización láser asistida por matriz (MALDI-TOF). La microbiota entérica fue diferente según el tratamiento. El GBH indujo un aumento de la diversidad bacteriana, mientras que la CIP produjo una reducción. Entre estos cambios, destaca la presencia de Yersinia spp. y Proteus spp. solo en el tratamiento con GBH. Además, en el tratamiento GBH-CIP se encontró una disminución en la prevalencia de Klebsiella spp. y Pseudomonas spp. en la microbiota intestinal de los renacuajos. Este es el primer informe sobre la alteración del contenido bacteriano intestinal de renacuajos de R. arenarum producido por dos contaminantes emergentes de preocupación. Demostramos que el renacuajo del sapo común se puede utilizar como un organismo modelo no convencional para el monitoreo de la contaminación ambiental. Estos hallazgos constituyen el primer paso para comprender cómo la presencia de GBH y CIP en aguas dulces puede representar una amenaza para la vida silvestre y la salud humana a través de la disbiosis entérica asociada al efecto sobre la comunidad bacteriana.The high load of agrochemicals and antibiotics present in agricultural aquatic environments represents a risk for wildlife. Since enteric bacteria, which play a key role in the physiological functioning of their hosts, are sensitive to a wide variety of pollutants, their study allows to evaluate the health of organisms. This study aimed to evaluate the effects of commercial formulations of a glyphosate-based herbicide (GBH) and the antibiotic ciprofloxacin (CIP), individually and in mixture, on the bacterial diversity of the intestinal content of common toad (Rhinella arenarum) tadpoles. The diversity of cultivable fast-growing bacteria with low nutritional requirements was evaluated using classic microbiological tests and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry identification. Bacterial diversity varied among treatments. Taxa diversity increased in the GBH-treated group but decreased in the CIP-treated group. Remarkably, Yersinia spp. and Proteus spp. were only found in the GBH-treated group. The prevalence of Klebsiella spp. and Pseudomonas spp. decreased in the intestinal microbiota of the GBH-CIP-treated group. To our knowledge, this is the first report on the alteration of cultivable enteric bacteria of autochthonous tadpoles due to two pollutants of emerging concern. Our results demonstrate that R. arenarum tadpoles can be used as non-conventional model organisms for environmental pollution monitoring. Our preliminary findings would contribute to understanding how the presence of GBH and CIP in freshwaters may represent a threat to wildlife and human health by causing enteric dysbiosis of part of the bacterial community.Fil: Cuzziol Boccioni, Ana Paula. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Laboratorio de Ecotoxicologia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: García Effron, Guillermo. Universidad Nacional del Litoral; ArgentinaFil: Peltzer, Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Laboratorio de Ecotoxicologia; ArgentinaFil: Lajmanovich, Rafael Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Laboratorio de Ecotoxicologia; Argentin

Propiedades microbiológicas en un suelo de la Patagonia Argentina bajo la influencia de diferentes especies forestales

191-197In forest systems, leaf litter is usually the main source of nutrients for the vegetation and microorganisms. Leaf litter composition may vary according to the biomass and quality of the materials produced by different species. Tree roots affect the development and activity of microflora. The aim of this study was to evaluate the influence of two implanted tree species, with different composition of plant residues, on the soil characteristics. The soil is classified as an Andisol and is located in the Trevelín Forest Station, Chubut, Argentina, Lat. 43° S, Long 71°31' West. The samples were extracted from the surface soil of two forest plots with a dominant species each: Radiata Pine (Pinus radiata D. Don.) and European Oak (Quercus robur). The following analytical determinations were carried out on soil samples: organic carbon, microbial respiration, deshydrogenase activity, microbial count (bacterial communities, amilolytic, actinomyces and fungi) as well as the functional diversity of associated microbial communities. The Shannon-Weaver index (H) was calculated. The soil under Oak trees showed the highest values of microbial respiration, organic carbon and amilolytic and bacterial community counts. No significant differences were found in deshydrogenase activity, actinomyces and fungi counts between species. The principal component analysis showed significant variations in the physiology of microbial communities associated with these two tree species. The microorganism H diversity index was significantly greater for Pine. Plants residues of broadleaf species such as Oak have lower contents of recalcitrant substances than conifer species, and this fact, associated with the higher development of total and amylolytic bacteria, could result in an increase in the decomposition of plant residues in the soil and in higher organic carbon and microbial respiration values

Multicenter Study of Method-Dependent Epidemiological Cutoff Values for Detection of Resistance in Candida spp. and Aspergillus spp. to Amphotericin B and Echinocandins for the Etest Agar Diffusion Method

BSTRACT Method-dependent Etest epidemiological cutoff values (ECVs) are not available for susceptibility testing of either Candida or Aspergillus species with amphotericin B or echinocandins. In addition, reference caspofungin MICs for Candida spp. are unreliable. Candida and Aspergillus species wild-type (WT) Etest MIC distributions (microorganisms in a species-drug combination with no detectable phenotypic resistance) were established for 4,341 Candida albicans, 113 C. dubliniensis, 1,683 C. glabrata species complex (SC), 709 C. krusei, 767 C. parapsilosis SC, 796 C. tropicalis, 1,637 Aspergillus fumigatus SC, 238 A. flavus SC, 321 A. niger SC, and 247 A. terreus SC isolates. Etest MICs from 15 laboratories (in Argentina, Europe, Mexico, South Africa, and the United States) were pooled to establish Etest ECVs. Anidulafungin, caspofungin, micafungin, and amphotericin B ECVs (in micrograms per milliliter) encompassing �97.5% of the statistically modeled population were 0.016, 0.5, 0.03, and 1 for C. albicans; 0.03, 1, 0.03, and 2 for C. glabrata SC; 0.06, 1, 0.25, and 4 for C. krusei; 8, 4, 2, and 2 forC. parapsilosis SC; and 0.03, 1, 0.12, and 2 for C. tropicalis. The amphotericin B ECV was 0.25 � g/ml for C. dubliniensis and 2, 8, 2, and 16 � g/ml for the complexes of A. fumigatus, A. flavus, A. niger, and A. terreus, respectively. While anidulafung in Etest ECVs classified 92% of the Candida fks mutants evaluated as non-WT, the performance was lower for caspofungin (75%) and micafungin (84%) cutoffs. Finally, although anidulafungin (as an echinocandin surrogate susceptibility marker) and amphotericin B ECVs should identify Candida and Aspergillus isolates with reduced susceptibility to these agents using the Etest, these ECVs will not categorize a fungal isolate as susceptible or resistant, as breakpoints do. KEYWORDS ECVs, Etest ECVs, Etest MICs Candida, Etest MICs Aspergillus, WT isolates, amphotericin B resistance, antifungal resistance, echinocandin resistance, non-WT, susceptibility marke

Posaconazole MIC Distributions for Aspergillus fumigatus Species Complex by Four Methods: Impact of cyp51A Mutations on Estimation of Epidemiological Cutoff Values

ABSTRACT Estimating epidemiological cutoff endpoints (ECVs/ECOFFS) may be hindered by the overlap of MICs for mutant and nonmutant strains (strains harboring or not harboring mutations, respectively). Posaconazole MIC distributions for the Aspergillus fumigatus species complex were collected from 26 laboratories (in Australia, Canada, Europe, India, South and North America, and Taiwan) and published studies. Distributions that fulfilled CLSI criteria were pooled and ECVs were estimated. The sensitivity of three ECV analytical techniques (the ECOFFinder, normalized resistance interpretation [NRI], derivatization methods) to the inclusion of MICs for mutants was examined for three susceptibility testing methods (the CLSI, EUCAST, and Etest methods). The totals of posaconazole MICs for nonmutant isolates (isolates with no known cyp51A mutations) and mutant A. fumigatus isolates were as follows: by the CLSI method, 2,223 and 274, respectively; by the EUCAST method, 556 and 52, respectively; and by Etest, 1,365 and 29, respectively. MICs for 381 isolates with unknown mutational status were also evaluated with the Sensititre YeastOne system (SYO). We observed an overlap in posaconazole MICs among nonmutants and cyp51A mutants. At the commonly chosen percentage of the modeled wild-type population (97.5%), almost all ECVs remained the same when the MICs for nonmutant and mutant distributions were merged: ECOFFinder ECVs, 0.5 μg/ml for the CLSI method and 0.25 μg/ml for the EUCAST method and Etest; NRI ECVs, 0.5 μg/ml for all three methods. However, the ECOFFinder ECV for 95% of the nonmutant population by the CLSI method was 0.25 μg/ml. The tentative ECOFFinder ECV with SYO was 0.06 μg/ml (data from 3/8 laboratories). Derivatization ECVs with or without mutant inclusion were either 0.25 μg/ml (CLSI, EUCAST, Etest) or 0.06 μg/ml (SYO). It appears that ECV analytical techniques may not be vulnerable to overlap between presumptive wild-type isolates and cyp51A mutants when up to 11.6% of the estimated wild-type population includes mutants. KEYWORDS Aspergillus fumigatus, CLSI ECVs, ECVs, EUCAST ECVs, Etest, SYO, cyp51A mutants, posaconazole, triazole resistance, wild typ

Influence of Metalworking Defect in Knee Cruciate Ligament Screws as a Possible Origin of Infections.

AbstractA bone mucormycosis outbreak in a clinic of Paraná city after arthroscopic knee anterior cruciate ligament reconstruction forced to suspend these interventions. The main candidates as the infection source were the implanted screws. These screws were metallurgically analyzed. Microscopic defects like burrs and bending's were found within the screw's Allen heads. According to previous report Spector et al. (2013), these defects protect microorganisms and make difficult to eliminate them. The aim of this work was to demonstrate the implication of metal working defects in the titanium fixing screws as a reservoir of contaminated machining oil which could be the possible transmitting vehicle in the outbreak of bone mucormycosis. The genetic relatedness of the isolated strains was established, favoring the hypothesis that contaminated screws might be the infection sources

Identification of Brucella ovis exclusive genes in field isolates from Argentina

La brucelosis ovina es una enfermedad infecciosa de curso crónico causada en nuestro país por Brucella ovis (B. ovis). Esta enfermedad constituye un problema importante para la producción ovina del país, especialmente en Patagonia, debido a su amplia difusión a nivel de establecimientos y a las pérdidas económicas que genera. El control de la enfermedad se basa actualmente en el diagnóstico y posterior descarte de animales infectados.Brucellosis caused by Brucella ovis is one of the most important infectious diseases of sheep. The aim of this study was to determine the presence of genes both inside and outside the specific B. ovis pathogenicity island 1 (BOPI-1) in a large collection of field isolates of B. ovis and other Brucella spp. from Argentina. The BOV_A0500 gene from B. ovis BOPI-1 was identified in all 104 B. ovis isolates studied. The BOPI-1 complete sequence was found to be conserved in 10 B. ovis strains from the collection, for which whole genome sequencing was performed. The BOV_0198 gene, which is outside BOPI-1 and considered exclusive to B. ovis, showed 90–100% identity with genomic regions of B. ovis, B. melitensis, B. abortus, B. canis, B. suis, B. microti, B. ceti and B. pinnipedialis. The results demonstrate that BOPI-1 is the only exclusive genetic region of B. ovis and marine Brucella spp. and that it is highly conserved in B. ovis field isolates from Argentina.EEA BarilocheFil: Alvarez, Lucia Paula. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Grupo Salud Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: García Effron, Guillermo. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Robles, Carlos Alejandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Grupo Salud Animal; Argentin

Discriminación entre Candida dubliniensis y Candida albicans evaluando el morfotipo de las colonias en agar Sabouraud-trifeniltetrazolio

Candida dubliniensis is a germ tube and chlamydoconidia producing Candida species that may be misidentified as Candida albicans. Molecular-based methods are the most reliable techniques for C. albicans and C. dubliniensis differentiation. However, accurate, quick and inexpensive phenotypic tests are needed to be used in low-complexity mycology laboratories.Candida dubliniensis es una especie del género Candida capaz de producir tubos germinativos y clamidoconidios, y puede ser identificada erróneamente como Candida albicans. Las técnicas moleculares de identificación son consideradas las más específicas para diferenciar estas especies. Sin embargo, se siguen necesitando métodos exactos, rápidos y de bajo coste para ser utilizados en laboratorios de micología de baja complejidad.Fil: Gamarra, Soledad. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Mancilla, Estefanía. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Dudiuk, Catiana Beatriz. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Santa Fe. Unidad de Adm.territorial; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: García Effron, Guillermo. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentin

Aspergillus fumigatus intrinsic fluconazole resistance is due to the naturally occurring T301I substitution in Cyp51Ap

Aspergillus fumigatus intrinsic fluconazole resistance has been demonstrated to be linked to the CYP51A gene, although the precisemolecular mechanism has not been elucidated yet. Comparisons between A. fumigatus Cyp51Ap and Candida albicansErg11p sequences showed differences in amino acid residues already associated with fluconazole resistance in C. albicans. Theaim of this study was to analyze the role of natural polymorphism I301 in Aspergillus fumigatus Cyp51Ap in the intrinsic fluconazoleresistance phenotype of this pathogen. The I301 residue in A. fumigatus Cyp51Ap was replaced with a threonine (analogueto T315 at Candida albicans fluconazole-susceptible Erg11p) by changing one single nucleotide in CYP51A gene. Also, aCYP51A knockout strain was obtained using the same parental strain. Both mutants? antifungal susceptibilities were tested. TheI301T mutant exhibited a lower level of resistance to fluconazole (MIC, 20 g/ml) than the parental strain (MIC, 640 g/ml),while no changes in MIC were observed for other azole- and non-azole-based drugs. These data strongly implicate the A. fumigatusCyp51Ap I301 residue in the intrinsic resistance to fluconazole.Fil: Leonardelli, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Macedo, Daiana. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Dudiuk, Catiana Beatriz. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Cabeza, Matías Sebastián. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Gamarra, Maria Soledad. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: García-Effron, Guillermo. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentin

First clinical isolation of an azole-resistant aspergillus fumigatus isolate harboring a TR46 Y121F T289A mutation in South America

One of the most recently described Aspergillus fumigatus CYP51Amediated azole resistance mechanisms is TR46 Y121F T289A. Clinical A. fumigatus strains harboring these substitutions have been reported worldwide, with the exception of South America. We describe the first clinical A. fumigatus strain with this resistance mechanism isolated from an Argentinian patient. The strain was isolated in 2009 (1 year after the first-described mutant in United States), demonstrating that these alleles were scattered worldwide earlier than previously thought.Fil: Isla, Guillermina. Instituto Nacional de Enfermedades Infecciosas; ArgentinaFil: Leonardelli, Florencia. Universidad Nacional del Litoral; ArgentinaFil: Tiraboschi, Iris Nora. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Refojo, Nicolás. Universidad de Buenos Aires. Facultad de Medicina. Hospital de Clínicas General San Martín; ArgentinaFil: Hevia, Alejandra. Instituto Nacional de Enfermedades Infecciosas; ArgentinaFil: Vivot, Walter Oscar. Instituto Nacional de Enfermedades Infecciosas; ArgentinaFil: Szusz, Wanda. Instituto Nacional de Enfermedades Infecciosas; ArgentinaFil: Córdoba, Susana Beatríz. Instituto Nacional de Enfermedades Infecciosas; ArgentinaFil: García Effron, Guillermo. Instituto Nacional de Enfermedades Infecciosas; Argentin