Streptomycin as a selective agent to facilitate recovery and isolation of introduced and indigenous Sphingomonas from environmental samples

Abstract

Sphingomonas is an organism of major interest for the degradation of organic contaminants in soils and other environments. A medium based on the aminoglycoside antibiotic streptomycin (Sm) was developed, which, together with the yellow pigmentation of Sphingomonas, facilitated the detection, recovery and quantification of culturable Sphingomonas from soils. All 29 previously described bacterial strains belonging to 17 different Sphingomonas species were able to grow on mineral media containing 200 microg ml(-1) streptomycin, showing that the capacity to resist high concentrations of Sm is a common characteristic within Sphingomonas. Incorporation of Sm into the mineral medium led to a significant reduction in the background microbial population and a concomitant 100 times more sensitive detection of Sphingomonas inoculated in non-sterile soil matrices. The Sm-containing medium was used to examine a variety of hydrocarbon-contaminated soils for the presence and biodiversity of Sphingomonas. Incorporation of Sm in the medium led to a significant increase in the number of yellow-pigmented colonies. Comparison of contaminated and non-contaminated soils derived from the same site revealed colonization by culturable yellow-pigmented Sm-resistant bacteria of the polluted location solely. Both yellow and non-yellow-pigmented colonies were purified from plates containing glucose and Sm, and BOX-polymerase chain reaction (PCR) was used to sort out clonally related strains. Representative strains from the major BOX-PCR clusters were identified using FAME and partial 16S rRNA gene sequencing. Forty-eight of 58 Sm-resistant isolates were identified as Sphingomonas sp. Streptomycin-resistant Sphingomonas isolates generated BOX-PCR diversity patterns that were site dependent and represented different species mainly belonging to Sphingomonas subgroups containing species formerly designated as Sphingopyxis and Sphingobium. The ability to degrade phenanthrene was only found in a minority of the Sphingomonas isolates, which all originated from soils containing high phenanthrene concentrations.

A Novel β-Peptidyl Aminopeptidase (BapA) from Strain 3-2W4 Cleaves Peptide Bonds of Synthetic β-Tri- and β-Dipeptides

“…Analysis of cellular fatty acids also resulted in a pattern typical for the genus Sphingomonas. Just recently, resistance against streptomycin was described as a characteristic typical of Sphingomonas strains (40). When unclassified bacteria were included in the 16S rRNA gene sequence analysis, the closest match was strain Y2 with 100% identity (GenBank accession number AB084247).…”

Novosphingobium panipatense sp. nov. and Novosphingobium mathurense sp. nov., from oil-contaminated soil

“…T and SM117 T were selectively isolated from oil-contaminated soil on minimal salt medium (MSM) containing 200 mg streptomycin ml 21 (Mohn et al, 2006;Vanbroekhoven et al, 2004). For this purpose, 1 g soil was suspended in 5 ml saline water and the mixture was diluted 10-fold and plated on MSM medium .…”

Abundance and Community Structure of Sphingomonads in Leaf Residues and Nearby Bulk Soil

“…The presence or even the dominance of non-culturable sphingomonads in a variety of hydrocarbon-contaminated soils was suggested (14,34). Therefore, culture-independent analyses of sphingomonad communities are needed in order to reveal their behavior, distribution, and community structure in the field.…”