Diversity of Chromobacterium violaceum isolates from aquatic environments of state of Pará, Brazilian Amazon

Abstract

The present study intended to characterize the phenotypic and genetic diversity of Brazilian isolates of Chromobacterium violaceum from aquatic environments within the Amazon region. Nineteen isolates showed morphological properties of C. violaceum and the majority grew at 44°C. Low temperatures, in contrast, showed (Ueda et al. 1994, Melo et al. 2000 and anti-Mycobacterium tuberculosis (de Souza et al. 1999) activities. Other properties of C. violaceum include the production of cyanide (Michaels & Corpe 1965), the solubilization of gold (Faramarzi et al. 2004), the production of chitinolytic enzymes (Chernin et al. 1998), the synthesis of bioplastics (Steinbüchel et al. 1993) and environmental detoxification (Carepo et al. 2004). Although it is a valuable biotechnological resource, C. violaceum is a highly virulent opportunistic pathogen to humans and animals (Chen et al. 2003, Brito et al. 2004, de Siqueira et al. 2005). Recombinase A (RecA) is a multifunctional protein involved in general recombination, DNA repair and the SOS response, and it is highly conserved among eubacteria (Cox 2003). Several studies have shown that recA can be used as a molecular tool to study diversity within the Erwinia genus (Waleron et al. 2002), C. violaceum (Scholz et al. 2005, Ochrobactrum anthropi (Scholz et al. 2006) and the Burkholderia cepacia complex (Seo & Tsuchiya 2004), despite this gene's high degree of nucleotide diversity (Casati et al. 2004).The present study aims to further characterize the phenotypic and genetic diversity of Brazilian isolates of C. violaceum from aquatic environments within the Amazon region. MATERIALS AND METHODSIsolates from natural water resources and reference strain -Water samples were collected from domestic wells (less than 15 m in depth) in Barcarena (site 1) and in Vila Bonifácio, municipality of Bragança (site 2). Samples were also collected from rivers within the National Forest of Caxiuana (site 3) and in Belém, the capital of Pará (site 4).Nineteen isolates of C. violaceum were recovered from superficial and underground water using the Membrane Filter (MF) and Most Probable Number (MPN) technique, according to the Standard Methods for the Examination of Water and Wastewater (APHA 2005). The MF technique was performed by the filtraDiversity of C. violaceum in the Brazilian Amazon • LT Dall'Agnol et al. 679tion of water samples through a sterile cellulose ester membrane with 0.45 µm pores. Membranes were transferred to m-Endo-type media with incubation at 37ºC for 24 h. The MPN technique consisted of the inoculation of sequential volumes of water samples in Lactose Broth, followed by incubation at 37ºC for 24-48 h. The colonies that presented a violet pigmentation in the isolation medium were transferred to Agar Stock (storage medium). Prior to biochemical or molecular testing, isolates were grown in Nutrient Broth (NB) for 24 h at 37ºC and placed in Nutrient Agar to re-isolate and confirm the anoxic condition of the culture. The reference strain ATCC 12472 was obtained from the Br...

Isolation and Identification of Chromobacterium Sp. From Different Ecosystems

“…However, in citrate reduction and sugar fermentation studies some variations we found. Our results were similar with the study of Dall' Agnol et al [24]. Their study stated that larger phenotypic variation was observed in the citrate test and the oxidation/fermentation of sucrose.…”

Antifungal activity of violacein purified from a novel strain of Chromobacterium sp. NIIST (MTCC 5522)

“…Significant genetic variability within and between populations of Chromobacterium sp. have been reported earlier (Dall'Agnol et al, 2008).…”

Chromobacterium vaccinii sp. nov., isolated from native and cultivated cranberry (Vaccinium macrocarpon Ait.) bogs and irrigation ponds

“…The current edition of Bergey's Manual of Systematic Bacteriology lists only the type species, Chromobacterium violaceum (Gillis & Logan, 2005), but five additional species have been proposed since 2007, based on the recognition of significant genetic, metabolic and ecological differences: Chromobacterium subtsugae (Martin et al, 2007), C. aquaticum (Young et al, 2008), C. haemolyticum (Han et al, 2008), C. piscinae (Kämpfer et al, 2009) and C. pseudoviolaceum (Kämpfer et al, 2009). Detailed analyses of C. violaceum collected from geographically diverse tropical regions indicate that there is considerable genetic variability within Chromobacterium (Ponnusamy et al, 2011;Dall'Agnol et al, 2008;Lima-Bittencourt et al, 2007, 2011Barreto et al, 2008), a finding which tends to support the recognition of additional species within Chromobacterium, particularly if these differences can be tied to variations in vertebrate pathogenicity or significant differences in ecological adaptation (Vinatzer & Bull, 2009). …”