Characterization of high fluoride resistant Pseudomonas aeruginosa species isolated from water samples

Abstract

Groundwater fluoride contamination is one of the dominant toxicological environmental problems in India and overseas. Water samples were taken from Natham taluk and screened the fluoride resistant bacteria. Initially, Twenty- four fluoride resistant colonies were selected from 50 mM NaF supplemented LB agar plates. All isolates showed β-haemolysis on blood agar plates, confirmed as pathogens. Virulence factors (algD, plcH, toxA, gyrB, rhlC, lasB) and biofilm-forming genes (ppyR, pelA, pslA) were identified in these isolates by PCR analysis. Fluoride ion transporter ‘crcB’ was successfully amplified from these isolates by gene-specific PCR. Genus Pseudomonas and P. aeruginosa species-specific PCR analysis identified that all strains belonging to P. aeruginosa species. Besides, three high fluoride resistance strains were selected based on high fluoride resistance and confirmed as P. aeruginosa species by 16S rRNA sequencing and NCBI blast analysis.

Characterization of β-lactamase and virulence genes in Pseudomonas aeruginosa isolated from clinical, environmental and poultry sources in Bangladesh

“…PCR amplification of 16s_ Pseudo for the detection of Pseudomonas genus [ 23 ] and Pseudo_aeru for the detection of P . aeruginosa species [ 24 , 25 ] was performed on all phenotypically identified isolates of P . aeruginosa .…”

Characterization of β-lactamase and virulence genes inPseudomonas aeruginosaisolated from clinical, environmental and poultry sources in Bangladesh

“…To amplify the 16S rRNA sequences of Pseudomonas , a set previously designed primer set was used ( Table 1 ). PCR amplification of 16s_ Pseudo for the detection of Pseudomonas genus [21] and Pseudo_aeru for the detection of P. aeruginosa species [22, 23] was performed on all phenotypically identified isolates of P. aeruginosa . The PCR condition against these primer sets for the amplification of genus- and species-specific primers is shown in Table S2 .…”

“…Amplification of targeted DNA was carried out in a 20 µL reaction mixture, including 3 µL nuclease-free water, 10 µL 2X master mixture (Promega, Madison, WI, USA), 1 µL forward and reverse primers (for each) and 5 µL DNA template. PCR-positive controls consisted of P. aeruginosa genomic DNA previously confirmed for the target genes [21][22][23].…”

Pretreatments for Microbial Analysis and Evaluation of Hygiene of Wet Towels and Wet Wipes

“…It was heated in a heat block at 99°C for 10 min, centrifuged at 15,814×g for 3 min, and the supernatant was used as template DNA. The universal bacterial 16s rRNA primers 27F (5'-AGA GTT TGA TCM TGG CTC AG-3') and 1492R (5'-CGG TTA CCT TGT TAC GAC TT-3') 15 were used for identification with 16s rRNA sequencing. The PCR product mixture was amplified with modified cycling conditions [95°C for 4 min (initial denaturation), 30 cycles of 95°C for 30 sec (denaturation), 55°C for 40 sec (annealing), and 72°C for 1 min (extension)] using Rotor-Gene Q (Qiagen).…”

“…The PCR product mixture was amplified with modified cycling conditions [95°C for 4 min (initial denaturation), 30 cycles of 95°C for 30 sec (denaturation), 55°C for 40 sec (annealing), and 72°C for 1 min (extension)] using Rotor-Gene Q (Qiagen). 15 Fluorescencelabeled ddNTP was added to the PCR product using a BigDye Terminator v3.1 matrix standard kit (ThermoFisher Scientific). Sequencing was performed with Applied Biosystems 3730 DNA Analyzer (Thermo Fisher Scientific).…”