Agarose gel (1%) analysis of PCR product of SXT integrase from IDH isolates and their transconjugants.

<p>PCR products obtained using genomic DNA templates from clinical isolates or their transconjugants have been electrophoresed in different lanes as follows: Lane M : 1 kb ladder (Fermentas); Lane 1: Positive control <i>V.cholerae</i> O139 MO10; Lane 2: Recipient <i>E. coli</i> XL-1 Blue; Lanes 3 and 4: Negative controls of no DNA template and SXT-negative IDH02095 isolate respectively; Lanes 5 and 6 : IDH01572 (SXT-positive) isolate and its transconjugant respectively; Lanes 7 and 8 : IDH01738 (SXT-positive) isolate and its transconjugant respectively.</p

Antimicrobial susceptibility of IDH01572, IDH01738 and their transconjugants.

<p>The antibiotic names are as described in legend to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056477#pone-0056477-g001" target="_blank">figure 1</a>.</p><p>Bold face indicates the resistance traits from recipient XL-1Blue cells.</p

Antibiotic susceptibility profile of 119 clinical isolates from Kolkata, India, in 2009.

<p>AMP, Ampicillin; CHL, Chloramphenicol; CIP, Ciprofloxacin; COT, Co-Trimoxazole; GEN, Gentamicin; KAN, Kanamycin; NAL, Nalidixic Acid; NEO, Neomycin; NOR, Norfloxacin; PB, Polymixin B; STR, Streptomycin; SUL, Sulfisoxazole; TET, Tetracycline; TRI, Trimethoprim.</p

Cytotoxic effect of environmental isolates on HT29 cells.

<p>HT29 cells not treated with supernatant from any of the isolates (A); Cell rounding in HT29 cells treated with the supernatant from P1(B); Clumping of HT29 cells treated with the supernatant from Z2 (C); Bar diagram showing cytotoxic effect of culture supernatants from environmental isolates and the controls <i>V. cholerae</i> N16961 and <i>E. coli</i> DH5α in terms of percentage of dead cell (D). The experiment was independently performed three times. The error bars indicate standard deviation.</p

Rabbit ileal loop assay to assess entrotoxigenic activity.

<p>Pictorial view of rabbit ileal loop of different clinical and environmental strains (A). Analysis of fluid accumulation of different <i>V. cholerae</i> strains (B). Rabbit ileal loops were inoculated with 10⁸ CFU of each strain N16961 (O1 El tor, clinical), IDH02365(O1 El tor, clinical), Z2 (non-O1/non-O139, Environmental) and W3(non-O1/non-O139, Environmental) in PBS and incubated for 18 h. Results are expressed as fluid accumulation (FA) (in milliliters) per loop length (in centimeters). Shown are means ± standard deviations; <i>n</i> = 3.</p

Analysis of protease activity of the environmental isolates.

<p>SDS PAGE (10%) analysis of culture supernatants (A). Lane 1: Protein markers (Pageruler, Fermentas) with molecular mass in kDa indicated on the left; Lanes 2 to 8, culture supernatants of Z2, Z3, P2, W1, W3, <i>V. cholerae</i> N16961 and <i>E. coli</i> DH5α, respectively. Protease activity of culture supernatants on 1.5% Skim milk agar plate (B). Sample identity has been indicated near each well.</p