Aislamiento e identificación de bacterias cultivables de la zona de raíz de granada roja (Punica granatum) en un huerto del Tephé Ixmiquilpan, Hidalgo, México

La microbiota bacteriana asociada a la zona de raí z sintetiza a cidos orga nicos, fitohormonas, pe ptidos, pigmentos y metabolitos bioactivos. Se determino la Capacidad de Intercambio Catio nico (CIC) del suelo, textura y pH. Los aislamientos bacterianos se identificaron mediante pruebas morfolo gicas y el metabolismo a trave s del sistema API20NE para el ge nero Pseudomonas. El ana lisis molecular de PCR utilizando iniciadores universales para la secuenciacio n del gen 16s ARNr. La secuencia consenso resultante de cada una de los aislados se comparo con el programa nBLAST del Centro Nacional NCBI. El suelo resulto ser Franco-arenoso con un pH de ligeramente a moderadamente alcalino y una CICT de bajo a medio. Se identificaron las especies Bacillus subtilis, Erwinia bilingea en la zona de raí z y Pseudomonas soli, P. oryzihabitans, P. putida y P. chlororaphis como endofí ticas. P. soli ha sido descrita recientemente (2014), con nuevo registro. P. oryzihabitans, es nativa de la rizosfera.The bacterial microbiota associated with the root zone synthesizes organic acids, phytohormones, peptides, pigments and bioactive metabolites. The Cation Exchange Capacity (CEC) of the soil, texture and pH were determined. Bacterial isolates were identified by morphological tests and metabolism through the API20NE system for the genus Pseudomonas. Molecular PCR analysis using universal primers for 16s rRNA gene sequencing. The consensus sequence resulting from each of the isolates was compared with the NCBI National Center's nBLAST program. The soil was found to be sandy loam with a pH of slightly to moderately alkaline and a CICT of low to medium. The species Bacillus subtilis, Erwinia bilingea in the root zone and Pseudomonas soli, P. oryzihabitans, P. putida and P. chlororaphis were identified as endophytic. P. soli has been described recently (2014), with a new record. P. oryzihabitans, is native to the rhizosphere

Efecto antagónico de Pseudomonaschlororaphis y Pseudomonasoryzihabitans sobre bacterias de interés médico

Se realizó un estudio exploratorio para evaluar el efecto de Pseudomonas chlororaphis y Pseudomonas oryzihabitans aisladas de raíz de granada en el crecimiento de bacterias de interés médico. El género Pseudomonas se caracteriza por la capacidad de metabolizar sustratos, producir pigmentos y otros metabolitos secundarios con efecto biocida como mecanismo para el control de poblaciones microbianas. Las cepas de P. chlororaphis y P. oryzihabitans se cultivaron en caldo nutritivo a 30°C por siete y veintiún días. Posteriormente se tomaron alícuotas, se centrifugaron a 10,000 rpm/10 minutos, y se avaluó su efecto Staphylococcus aureus, Salmonella sp. y Bacillus subtilis mediante la prueba de Kirby-Bauer. Ambas cepas de Pseudomonas mostraron actividad inhibitoria en Staphylococcus aureus, Salmonella sp. y Bacillus subtilis.An exploratory study was carried out to evaluate the effect of Pseudomonas chlororaphis and Pseudomonas oryzihabitans isolated from pomegranate root on the growth of bacteria of medical interest. The genus Pseudomonas is characterized by the ability to metabolize substrates, produce pigments and other secondary metabolites with a biocidal effect as a mechanism for the control of microbial populations. The P. chlororaphis and P. oryzihabitans strains were cultured in nutrient broth at 30° C for seven and twenty-one days. Afterwards, aliquots were centrifuged at 10,000 rpm/10 minutes, and their effect was evaluated on Staphylococcus aureus, Salmonella sp. and Bacillus subtilis using the Kirby-Bauer test. Both strains of Pseudomonas showed inhibitory activity in Staphylococcus aureus, Salmonella sp. and Bacillus subtilis

International Society of Human and Animal Mycology (ISHAM)-ITS reference DNA barcoding database - the quality controlled standard tool for routine identification of human and animal pathogenic fungi

Human and animal fungal pathogens are a growing threat worldwide leading to emerging infections and creating new risks for established ones. There is a growing need for a rapid and accurate identification of pathogens to enable early diagnosis and targeted antifungal therapy. Morphological and biochemical identification methods are time-consuming and require trained experts. Alternatively, molecular methods, such as DNA barcoding, a powerful and easy tool for rapid monophasic identification, offer a practical approach for species identification and less demanding in terms of taxonomical expertise. However, its wide-spread use is still limited by a lack of quality-controlled reference databases and the evolving recognition and definition of new fungal species/complexes. An international consortium of medical mycology laboratories was formed aiming to establish a quality controlled ITS database under the umbrella of the ISHAM working group on "DNA barcoding of human and animal pathogenic fungi." A new database, containing 2800 ITS sequences representing 421 fungal species, providing the medical community with a freely accessible tool at http://www.isham.org and http://its.mycologylab.org/ to rapidly and reliably identify most agents of mycoses, was established. The generated sequences included in the new database were used to evaluate the variation and overall utility of the ITS region for the identification of pathogenic fungi at intra-and interspecies level. The average intraspecies variation ranged from 0 to 2.25%. This highlighted selected pathogenic fungal species, such as the dermatophytes and emerging yeast, for which additional molecular methods/genetic markers are required for their reliable identification from clinical and veterinary specimens.This study was supported by an National Health and Medical Research Council of Australia (NH&MRC) grant [#APP1031952] to W Meyer, S Chen, V Robert, and D Ellis; CNPq [350338/2000-0] and FAPERJ [E-26/103.157/2011] grants to RM Zancope-Oliveira; CNPq [308011/2010-4] and FAPESP [2007/08575-1] Fundacao de Amparo Pesquisa do Estado de So Paulo (FAPESP) grants to AL Colombo; PEst-OE/BIA/UI4050/2014 from Fundacao para a Ciencia e Tecnologia (FCT) to C Pais; the Belgian Science Policy Office (Belspo) to BCCM/IHEM; the MEXBOL program of CONACyT-Mexico, [ref. number: 1228961 to ML Taylor and [122481] to C Toriello; the Institut Pasteur and Institut de Veil le Sanitaire to F Dromer and D Garcia-Hermoso; and the grants from the Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq) and the Fundacao de Amparo a Pesquisa do Estado de Goias (FAPEG) to CM de Almeida Soares and JA Parente Rocha. I Arthur would like to thank G Cherian, A Higgins and the staff of the Molecular Diagnostics Laboratory, Division of Microbiology and Infectious Diseases, Path West, QEII Medial Centre. Dromer would like to thank for the technical help of the sequencing facility and specifically that of I, Diancourt, A-S Delannoy-Vieillard, J-M Thiberge (Genotyping of Pathogens and Public Health, Institut Pasteur). RM Zancope-Oliveira would like to thank the Genomic/DNA Sequencing Platform at Fundacao Oswaldo Cruz-PDTIS/FIOCRUZ [RPT01A], Brazil for the sequencing. B Robbertse and CL Schoch acknowledge support from the Intramural Research Program of the NIH, National Library of Medicine. T Sorrell's work is funded by the NH&MRC of Australia; she is a Sydney Medical School Foundation Fellow.info:eu-repo/semantics/publishedVersio

Biological activity of the mite Sancassania sp. (Acari: Acaridae) from bat guano associated with the pathogenic fungus Histoplasma capsulatum

Mites and the mammal pathogenic fungus Histoplasma capsulatum are the major components of bat guano microbiota. Interactions between mites and H. capsulatum were evaluated under laboratory conditions. Acarid mites, mainly Sancassania sp., were the most abundant microarthropod in the sampled guano of the Mexican bat Tadarida brasiliensis mexicana and, based on its morphology, Sancassania sp. was similar to the cosmopolitan species Sancassania sphaerogaster. The mycophagous and vectoring activities of this mite were tested for H. capsulatum and two other fungal species, Sporothrix schenckii (pathogenic) and Aspergillus sclerotiorum (non-pathogenic). S. ca. sphaerogaster was able to reproduce in H. capsulatum and S. schenckii colonies, multiplying in great numbers under controlled fungal mycelial-phase culture conditions. H. capsulatum colonies were completely destroyed after 14 days of in vitro interaction with mites. In contrast, S. ca. sphaerogaster did not reproduce in A. sclerotiorum cultures. S. ca. sphaerogaster was found vectoring H. capsulatum, but not the two other fungal species studied

Biological activity of the mite Sancassania (Acari: Acaridae) from bat guano associated with the pathogenic fungus Histoplasma capsulatum

Mites and the mammal pathogenic fungus Histoplasma capsulatum are the major components of bat guano microbiota. Interactions between mites and H. capsulatum were evaluated under laboratory conditions. Acarid mites, mainly Sancassania , were the most abundant microarthropod in the sampled guano of the Mexican bat Tadarida brasiliensis mexicana and, based on its morphology, Sancassania sp. was similar to the cosmopolitan species Sancassania sphaerogaster . The mycophagous and vectoring activities of this mite were tested for H. capsulatum and two other fungal species, Sporothrix schenckii (pathogenic) and Aspergillus sclerotiorum (non-pathogenic). S. ca. sphaerogaster was able to reproduce in H. capsulatum and S. schenckii colonies, multiplying in great numbers under controlled fungal mycelial-phase culture conditions. H. capsulatum colonies were completely destroyed after 14 days of in vitro interaction with mites. In contrast, S. ca. sphaerogaster did not reproduce in A. sclerotiorum cultures. S. ca. sphaerogaster was found vectoring H. capsulatum, but not the two other fungal species studied

Profile of cytokines in the lungs of BALB/c mice after intra-nasal infection with Histoplasma capsulatum mycelial propagules

Fil: Sahaza, Jorge H. Laboratorio de Inmunología de Hongos, Unidad de Micología, Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM); México.Fil: Suárez-Alvarez, Roberto. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Micología; Argentina.Fil: Estrada-Bárcenas, Daniel Alfonso. Laboratorio de Inmunología de Hongos, Unidad de Micología, Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM); México.Fil: Pérez-Torres, Armando. Laboratorio de Filogenia del Sistema Inmune de Piel y Mucosas, Departamento de Biología Celular y Tisular, Facultad de Medicina, UNAM; México.Fil: Taylor, Maria Lucia. Laboratorio de Inmunología de Hongos, Unidad de Micología, Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM); México.The host pulmonary response to the fungus Histoplasma capsulatum was evaluated, through the profile of cytokines detected by the MagPix magnetic beads platform in lung homogenates and by lung-granulomas formation, from mice intra-nasally infected with mycelial propagules (M-phase) of two virulent H. capsulatum strains, EH-46 and G-217B. Results highlight that mice lung inflammatory response depends on the H. capsulatum strain used, during the first step of the fungal infection. IL-1β and TNF-α increased their concentrations in mice infected with both strains. The highest levels of IL-6, IL-17, and IL-23 were found in EH-46-infected mice, whereas levels of IL-22 were variable at all post-infection times for both strains. Significant increases of IL-12, IFN-γ, IL-4, and IL-10 were associated to EH-46-infected mice. Histological lung findings from EH-46-infected mice revealed incipient and numerous well-developed granulomas, distributed in lung-lobes at the 14th and the 21st days after infection, according to cytokine profiles

Histoplasma capsulatum Isolated from Tadarida brasiliensis Bats Captured in Mexico Form a Sister Group to North American Class 2 Clade.

Histoplasma capsulatum is a dimorphic fungus associated with respiratory and systemic infections in mammalian hosts that have inhaled infective mycelial propagules. A phylogenetic reconstruction of this pathogen, using partial sequences of arf, H-anti, ole1, and tub1 protein-coding genes, proposed that H. capsulatum has at least 11 phylogenetic species, highlighting a clade (BAC1) comprising three H. capsulatum isolates from infected bats captured in Mexico. Here, relationships for each individual locus and the concatenated coding regions of these genes were inferred using parsimony, maximum likelihood, and Bayesian inference methods. Coalescent-based analyses, a concatenated sequence-types (CSTs) network, and nucleotide diversities were also evaluated. The results suggest that six H. capsulatum isolates from the migratory bat Tadarida brasiliensis together with one isolate from a Mormoops megalophylla bat support a NAm 3 clade, replacing the formerly reported BAC1 clade. In addition, three H. capsulatum isolates from T. brasiliensis were classified as lineages. The concatenated sequence analyses and the CSTs network validate these findings, suggesting that NAm 3 is related to the North American class 2 clade and that both clades could share a recent common ancestor. Our results provide original information on the geographic distribution, genetic diversity, and host specificity of H. capsulatum

Genetic diversity of Histoplasma capsulatum isolated from infected bats randomly captured in Mexico, Brazil, and Argentina, using the polymorphism of (GA)(n) microsatellite and its flanking regions

Fil: Taylor, María Lucía. Universidad Nacional Autónoma de México. Facultad de Medicina. Departamento de Microbiología-Parasitología; México.Fil: Hernández-García, Lorena. Universidad Nacional Autónoma de México. Facultad de Medicina. Departamento de Microbiología-Parasitología; México.Fil: Estrada-Bárcenas, Daniel. Universidad Nacional Autónoma de México. Facultad de Medicina. Departamento de Microbiología-Parasitología; México.Fil: Salas-Lizana, Rodolfo. Universidad Nacional Autónoma de México. Instituto de Ecología. Departamento de Ecología Evolutiva; México.Fil: Zancopé-Oliveira, Rosely M. Fundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Serviço de Micología-Setor de Imunodiagnóstico; Brasil.Fil: García de la Cruz, Saúl. Universidad Nacional Autónoma de México. Facultad de Medicina. Departamento de Microbiología-Parasitología; México.Fil: Galvão-Dias, Maria A. Centro de Controle de Zoonoses de Sao Paulo; Brasil.Fil: Curiel-Quesada, Everardo. Instituto Politécnico Nacional. Departamento de Bioquímica. Escuela Nacional de Ciencias Biológicas; México.Fil: Canteros, Cristina E. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bioquímica; Argentina.Fil: Bojórquez-Torres, Georgina. Universidad Nacional Autónoma de México. Facultad de Medicina. Departamento de Microbiología-Parasitología; México.Fil: Bogard-Fuentes, Carlos A. Universidad Nacional Autónoma de México. Facultad de Medicina. Departamento de Microbiología-Parasitología; México.Fil: Zamora-Tehozol, Erick. Universidad Nacional Autónoma de México. Facultad de Medicina. Departamento de Microbiología-Parasitología; México.The genetic diversity of 47 Histoplasma capsulatum isolates from infected bats captured in Mexico, Brazil, and Argentina was studied, using sequence polymorphism of a 240-nucleotides (nt) fragment, which includes the (GA)(n) length microsatellite and its flanking regions within the HSP60 gene. Three human clinical strains were used as geographic references. Based on phylogenetic analyses of 240-nt fragments achieved, the relationships among H. capsulatum isolates were resolved using neighbour-joining and maximum parsimony methods. The tree topologies obtained by both methods were identical and highlighted two major clusters of isolates. Cluster I had three sub-clusters (Ia, Ib, and Ic), all of which contained Mexican H. capsulatum samples, while cluster II consisted of samples from Brazil and Argentina. Sub-cluster Ia included only fungal isolates from the migratory bat Tadarida brasiliensis. An average DNA mutation rate of 2.39 × 10(-9) substitutions per site per year was estimated for the 240-nt fragment for all H. capsulatum isolates. Nucleotide diversity analysis of the (GA)(n) and flanking regions from fungal isolates of each cluster and sub-cluster underscored the high similarity of cluster II (Brazil and Argentina), sub-clusters Ib, and Ic (Mexico). According to the genetic distances among isolates, a network of the 240-nt fragment was graphically represented by (GA)(n) length haplotype. This network showed an association between genetic variation and both the geographic distribution and the ecotype dispersion of H. capsulatum, which are related to the migratory behaviour of the infected bats studied