Diversity among strains of Pseudomonas aeruginosa from manure and soil, evaluated by multiple locus variable number tandem repeat analysis and antibiotic resistance profiles

Abstract

The results of a multiple locus variable number of tandem repeat (VNTR) analysis (MLVA)-based study designed to understand the genetic diversity of soil and manure-borne Pseudomonas aeruginosa isolates, and the relationship between these isolates and a set of clinical and environmental isolates, are hereby reported. Fifteen described VNTR markers were first selected, and 62 isolates recovered from agricultural and industrial soils in France and Burkina Faso, and from cattle and horse manure, along with 26 snake-related isolates and 17 environmental and clinical isolates from international collections, were genotyped. Following a comparison with previously published 9-marker MLVA schemes, an optimal 13-marker MLVA scheme (MLVA13-Lyon) was identified that was found to be the most efficient, as it showed high typability (90%) and high discriminatory power (0.987). A comparison of MLVA with PFGE for typing of the snake-related isolates confirmed the MLVA13-Lyon scheme to be a robust method for quickly discriminating and inferring genetic relatedness among environmental isolates. The 62 isolates displayed wide diversity, since 41 MLVA types (i.e. MTs) were observed, with 26 MTs clustered in 10 MLVA clonal complexes (MCs). Three and eight MCs were found among soil and manure isolates, respectively. Only one MC contained both soil and manure-borne isolates. No common MC was observed between soil and manure-borne isolates and the snake-related or environmental and clinical isolates. Antibiotic resistance profiles were performed to determine a potential link between resistance properties and the selective pressure that might be present in the various habitats. Except for four soil and manure isolates resistant to ticarcillin and ticarcillin/clavulanic acid and one isolate from a hydrocarbon-contaminated soil resistant to imipenem, all environmental isolates showed wild-type antibiotic profiles.

Low occurrence of Pseudomonas aeruginosa in agricultural soils with and without organic amendment

“…Data showed that those isolates had different MLVA types and differed from other environmental and clinical isolates (Youenou et al, 2014) suggesting that the detection of P. aeruginosa in the soil samples from Burkina Faso was not due to a laboratory contamination.…”

Toxicity assessment and microbial response to soil antibiotic exposure: differences between individual and mixed antibiotics

“…Among the species with signicant changes in different antibiotic treatments, Pseudomonas is the most well-studied, and many previous studies have found that the abundance of Pseudomonas changed with soil antibiotic amendments and showed antibiotic resistence. [98][99][100][101][102][103] A recent study demonstrates that Pseudomonas putida could actively form biolms to protect the bacterial population under tetracycline stress. 104 Pirellula and Gemmata belong to phylum Planctomycetes, which were reported to demonstrate broad-spectrum antibiotic resistance.…”

EsrC, an envelope stress‐regulated repressor of the mexCD‐oprJ multidrug efflux operon in Pseudomonas aeruginosa

“…Pseudomonas aeruginosa is an opportunistic human pathogen that occurs naturally in a variety of habitats including soil (Green et al ., ; Youenou et al ., ) and, especially, water, both fresh (Pellett et al ., ; Römling et al ., ; Aoi et al ., ; Howard et al ., ; Pirnay et al ., ; Khan et al ., ; Nonaka et al ., ; Selezska et al ., ) and marine (Römling et al ., ; Howard et al ., ; Kimata et al ., ; Nonaka et al ., ), often at sites of pollution (de Vicente et al ., ; Pirnay et al ., ) and in wastewater (Ringen and Drake, ; Ziegert and Stelzer, ; Howard et al ., ). In these environments, the organism will encounter a variety of stresses, naturally derived [e.g.…”

“…oxidative stress from reactive oxygen species generated by photochemical processes in aquatic ecosystems (Zepp et al ., ; Baxter and Carey, ; Cooper et al ., ) and nitrosative stress as a result of NO generation during the process of denitrification by this and other soil/aquatic denitrifying organisms (St John and Hollocher, ; Conrad, ; Bothe et al ., )] or promoted by environmental pollution/contamination [e.g., oxidative stress driven by heavy metals (Teitzel et al ., ; Barras and Fontecave, ; Macomber and Hausinger, ) and nitrosative stress driven by excess nitrogen from farming and other human activities (Fields, )]. Intriguingly, these stresses stimulate responses in P. aeruginosa that drive antibiotic resistance (Fraud and Poole, ; Fetar et al ., ), an interesting observation given the not uncommon occurrence of antibiotic‐resistant P. aeruginosa in natural, including soil (Pitondo‐Silva et al ., ; Youenou et al ., ) and aquatic (de Vicente et al ., ; Pirnay et al ., ; Khan et al ., ; Nonaka et al ., ; Suzuki et al ., ; Zanetti et al ., ) environments. The antibiotic efflux systems MexXY‐OprM and MexEF‐OprN, for example, are inducible by oxidative (Fraud and Poole, ) and nitrosative (Fetar et al ., ) stress, respectively, with these stresses selecting for efflux pump‐expressing antibiotic‐resistant mutants in vitro (Fraud and Poole, ; Fetar et al ., ).…”

“…triclosan (Chuanchuen et al ., ) and chlorhexidine (CHX) (Fraud et al ., )], organic solvents (Li et al ., ) and dyes and detergents (Srikumar et al ., 1997; 1998) and, so, its expression promotes multidrug resistance in this organism (Poole et al ., ; Chuanchuen et al ., ; Mulet et al ., ). The induction of mexCD‐oprJ by membrane‐damaging organic solvents/aromatic hydrocarbons (Fraud et al ., ) and its ability to promote solvent tolerance in vitro (Li et al ., ) may explain, in some instances at least, the presence of P. aeruginosa isolates at environmental sites of hydrocarbon contamination (Kaszab et al ., ; Youenou et al ., ) and the multidrug resistance of these isolates (Kaszab et al ., ; Youenou et al ., ).…”