Bioprospecting for culturable actinobacteria with antimicrobial properties isolated from rivers in Colombian Orinoquia

Purpose: To Isolate and characterize Actinobacteria with antimicrobial activity from Guaviare River (Colombia). Methods: Water and sediment samples were collected from Guaviare River. Direct plating, heat and CaCO3 methods were used to isolate Actinobacteria. Six bacterial strains were tested using T-Streak method: Escherichia coli ATCC 23724, Staphylococus aureus ATCC 25923, Acinetobacter baumannii ATCC 19606, Bacillus subtilis ATCC 21556, Klebsiella pneumoniae ATCC 700603, Chromobacterium violaceum ATCC 31532. Strains of Fusarium sp. H24, Trichoderma harzianum H5 and Colletotrichum gloeosporioides were tested using Kirby-Bauer method. Isolates with high antimicrobial activity were selected for further taxonomic identification. Results: A total of 374 actinobacteria isolates were obtained. Seven isolates exhibited high antimicrobial activity (p < 0.05) and were confirmed as members of Streptomycetaceae family. Of these, three isolates showed differential phenotypic and genotypic profiles, indicating that they may represent new species. Conclusions: To date, this is the first study of this type in Colombian Orinoquia and indicates that this promising source of Actinobacteria from aquatic sediments with the ability to produce antimicrobial secondary metabolites.

Book of abstracts...

2016

Wheat blast and its effect on technological quality of grain from two sowing dates.

2016

Efficacy of fungicides for the control of wheat blast: collaborative tests results.

2016

Fine phenotyping unlocks wheat mechanisms of reaction to Magnaporthe oryzae.

2016

A systemic approach to incorporate the highest levels of FHB resistance with agronomic merit and quality in wheat.

2016

Beech bark disease distribution and resistance in Michigan and fungal endophyte ecology of resistant and susceptible beech (Fagus grandifolia Ehrh.)

Beech bark disease (BBD), a non-native association of the fungal pathogen Neonectria faginata and the beech scale insect Cryptococcus fagisuga, has dramatically affected American beech within North American forests. To monitor the spread and effects of BBD in Michigan, a network of forest health monitoring plots was established in 2001 following the disease discovery in Ludington State Park (Mason County). Forest health canopy condition and basic forestry measurements including basal area were reassessed on beech trees in these plots in 2011 and 2012. The influence of bark-inhabiting fungal endophytes on BBD resistance was investigated by collecting cambium tissue from apparently resistant and susceptible beech. Vigor rating showed significant influences of BBD in sample beech resulting in reduced health and substantiated by significant increases of dead beech basal area over time. C. fagisuga distribution was found to be spatially clustered and widespread in the 22 counties in Michigan's Lower Peninsula which contained monitoring plots. Neonectria has been found in Emmet, Cheboygan and Wexford in the Lower Peninsula which may coincide with additional BBD introduction locations. Surveys for BBD resistance resulted in five apparently resistant beech which were added to a BBD resistance database. The most frequently isolated endophytes from cambium tissue were identified by DNA sequencing primarily as Deuteromycetes and Ascomycetes including Chaetomium globosum, Neohendersonia kickxii and Fusarium flocciferum. N. faginata in antagonism trials showed significant growth reduction when paired with three beech fungal endophytes. The results of the antagonism trial and decay tests indicate that N. faginata may be a relatively poor competitor in vivo with limited ability to degrade cellulose.

Descripción del análisis microbológico realizado mediante maldi-tof en muestras de la fundación santa fe de Bogotá.

Introducción: La rápida detección e identificación bacteriana es fundamental para el manejo de los pacientes críticos que presentan una patología infecciosa, esto requiere de métodos rápidos para el inicio de un correcto tratamiento. En Colombia se usan pruebas microbiología convencional. No hay estudios de espectrofotometría de masas en análisis de muestras de pacientes críticos en Colombia. Objetivo general: Describir la experiencia del análisis microbiológico mediante la tecnología MALDI-TOF MS en muestras tomadas en la Fundación Santa Fe de Bogotá. Materiales y Métodos: Entre junio y julio de 2013, se analizaron 147 aislamientos bacterianos de muestras clínicas, las cuales fueron procesadas previamente por medio del sistema VITEK II. Los aislamientos correspondieron a 88 hemocultivos (60%), 28 urocultivos (19%), y otros cultivos 31 (21%). Resultados: Se obtuvieron 147 aislamientos con identificación adecuada a nivel de género y/o especie así: en el 88.4% (130 muestras) a nivel de género y especie, con una concordancia del 100% comparado con el sistema VITEK II. El porcentaje de identificación fue de 66% en el grupo de bacilos gram negativos no fermentadores, 96% en enterobacterias, 100% en gérmenes fastidiosos, 92% en cocos gram positivos, 100% bacilos gram negativos móviles y 100% en levaduras. No se encontró ninguna concordancia en bacilos gram positivos y gérmenes del genero Aggregatibacter. Conclusiones: El MALDI-TOF es una prueba rápida para la identificación microbiológica de género y especie que concuerda con los resultados obtenidos de manera convencional. Faltan estudios para hacer del MALDI-TOF MS la prueba oro en identificación de gérmenes.

DETECTION OF SPORES AND HYPHAE OF ARTWORKS’ BIODETERIOGENIC FILAMENTOUS FUNGI BY RNA-FISH

Filamentous fungi are a threat to the conservation of Cultural Heritage. Thus, detection and identification of viable filamentous fungi are crucial for applying adequate Safeguard measures. RNA-FISH protocols have been previously applied with this aim in Cultural Heritage samples. However, only hyphae detection was reported in the literature, even if spores and conidia are not only a potential risk to Cultural Heritage but can also be harmful for the health of visitors, curators and restorers. Thus, the aim of this work was to evaluate various permeabilizing strategies for their application in the detection of spores/conidia and hyphae of artworks’ biodeteriogenic filamentous fungi by RNA-FISH. Besides of this, the influence of cell aging on the success of the technique and on the development of fungal autofluorescence (that could hamper the RNA-FISH signal detection) were also investigated. Five common biodeteriogenic filamentous fungi species isolated from biodegradated artworks were used as biological model: Aspergillus niger, Cladosporium sp, Fusarium sp, Penicillium sp. and Exophialia sp. Fungal autofluorescence was only detected in cells harvested from Fusarium sp, and Exophialia sp. old cultures, being aging-dependent. However, it was weak enough to allow autofluorescence/RNA-FISH signals distinction. Thus, autofluorescence was not a limitation for the application of RNA-FISH for detection of the taxa investigated. All the permeabilization strategies tested allowed to detect fungal cells from young cultures by RNA-FISH. However, only the combination of paraformaldehyde with Triton X-100 allowed the detection of conidia/spores and hyphae of old filamentous fungi. All the permeabilization strategies failed in the Aspergillus niger conidia/spores staining, which are known to be particularly difficult to permeabilize. But, even in spite of this, the application of this permeabilization method increased the analytical potential of RNA FISH in Cultural Heritage biodeterioration. Whereas much work is required to validate this RNA-FISH approach for its application in real samples from Cultural Heritage it could represent an important advance for the detection, not only of hyphae but also of spores and conidia of various filamentous fungi taxa by RNA-FISH.

Flowchart for the production of perithecia of Gibberella zeae in wheat grains.

2016

Methodology to determine the number of infection points of Gibberella zeae in wheat spikes.

2016

Effects of sowing dates and wheat genetic resistance in FHB control.

2016

Effects of baking process on mycotoxin content in whole and white breads.

2016

Seleção de bactérias endofíticas com ação antagônica a fitopatógenos.

Muitos métodos rápidos e eficientes de seleção de agentes de biocontrole de fitopatógenos tem sido utilizados, visando reduzir tempo e custo dispendido em testes de campo. Neste trabalho realizou-se uma seleção de isolados endofíticos com potencial de uso no biocontrole de fitopatógenos em testes de antagonismo in vitro. De um total de 95 isolados de bactérias endofíticas do milho, seis foram selecionados quanto à inibição a Pythium aphanidermatum. A essa seleção, foram incluídos um isolado de Bacillus subtilis 0G, Bacillus lentimorbus e Streptomyces sp., para verificação de antagonismo a Rhizoctonia solani, Fusarium moniliforme, Sclerotium rolfsii e Exserohilum turcicum. Verificou-se que os endofíticos B. subtilis 0G, B. lentimorbus e Streptomyces sp., apresentaram ação antagônica superior aos demais, com taxas de inibição entre 32,0% e 53,8%. Dentre os endofíticos do milho, Bacillus agaradhaerens foi o que mais se destacou, com taxas de inibição variando entre 43,7% e 52,3% e indicando uma inespecificidade de ação. Este estudo, embora preliminar, permite vislumbrar a utilização desses endofíticos na supressão de doenças em diferentes sistemas patógeno-hospedeiro em testes subseqüentes, sob condições de casa-de-vegetação e a campo.

Fungos associados às podridões pós-colheita em acerola.

A aceroleira é uma das principais culturas produzidas no Submédio do Vale do São Francisco, sendo importante fonte de renda na região. O fruto é rico em vitamina C e muito utilizado na indústria de sucos. As podridões resultantes da atividade de patógenos ocasionam graves perdas na pós-colheita de acerolas. Desta forma, o objetivo deste trabalho foi isolar e identificar os fungos associados às podridões em acerola em diferentes cultivares e estádios de maturação. Os frutos foram coletadas das cultivares Flor Branca e Junco, em quatro estádios de maturação (0%, 1-25%, 25-75% e 75%-100% de coloração vermelha da casca). Posteriormente, os mesmos foram colocados em câmara úmida por 48 horas, em temperatura de 25o C e avaliados quanto à incidência de fungos causadores de podridões pós-colheita. Os fungos Aspergillus e Mucor predominaram com 96% e 100% para as cultivares Junco e Flor Branca, respectivamente. Alguns fungos só se desenvolvem quando os frutos atingem 75% de maturação, tais como Alternaria, Fusarium e Lasiodiplodia. Aspergillus e Mucor são encontrados independentemente do estádio de maturação do fruto.