Nalidixic Acid-susceptible Strains Research Articles

Efficacy of Ciprofloxacin for Treatment of Cholera Associated with Diminished Susceptibility to Ciprofloxacin to Vibrio cholerae O1.

ObjectiveWe identified a poor clinical response to treatment of cholera with a single 1 g dose of ciprofloxacin, a standard treatment for cholera.MethodsTo determine reasons for the poor response and better therapeutic approaches we examined the minimal inhibitor concentration (MIC, n = 275) and disc-diffusion zone sizes (n = 205) for ciprofloxacin and nalidixic acid of V. cholerae O1 strains isolated in Bangladesh from 1994 to 2012, and reexamined data from 161patients infected with Vibrio cholerae O1 recruited in four clinical trials who received single- or multiple-dose ciprofloxacin for treatment of cholera and compared their clinical response to the V. cholerae O1 susceptibility.ResultsAlthough all 275 isolates of V. cholerae O1 remained susceptible to ciprofloxacin using standard MIC and disc-diffusion thresholds, the MIC90 to ciprofloxacin increased from 0.010 in 1994 to 0.475 μgm/ml in 2012. Isolates became frankly resistant to nalidixic with the MIC90 increasing from 21 μgm/ml in 1994 to >256 μgm/ml and 166 of 205 isolates from 1994 to 2005 being frankly resistant using disc-diffusion testing. Isolates resistant to nalidixic acid by disc-diffusion testing had a median ciprofloxacin MIC of 0.190 μgm/ml (10th-90th centiles 0.022 to 0.380); nalidixic acid-susceptible isolates had a median ciprofloxacin MIC of 0.002 (0.002 to 0.012).The rate of clinical success with single-dose ciprofloxacin treatment for nalidixic acid-susceptible strains was 94% (61 of 65 patients) and bacteriologic success 97% (63/65) compared to 18% (12/67) and 8% (5/67) respectively with nalidixic acid-resistant strains (P<0.001 for both comparisons). Multiple-dose treatment with ciprofloxacin had 86% and 100% clinical and bacteriologic success rates respectively in patients infected with nalidixic acid-susceptible strains of V. cholerae O1 compared to clinical success 67% and bacteriologic success 60% with nalidixic acid-resistant strains.ConclusionsSingle-dose ciprofloxacin is not effective for treating cholera caused by V. cholerae O1 with diminished susceptibility to ciprofloxacin, and nalidixic acid disc-diffusion testing effectively screens for such isolates.

Nalidixic Acid-Resistant Strains of Salmonella Showing Decreased Susceptibility to Fluoroquinolones in the Midwestern Region of the Republic of Ireland Due to Mutations in the gyrA Gene

Nalidixic Acid-Resistant Strains of Salmonella Showing Decreased Susceptibility to Fluoroquinolones in the Midwestern Region of the Republic of Ireland Due to Mutations in the <i>gyrA</i> Gene

Plasmid-mediated quinolone resistance in expanded spectrum beta lactamase producing enterobacteriaceae in Morocco

Although independently acquired, plasmid-mediated quinolone resistance appears to be linked with extended-spectrum or AmpC-type beta-lactamases. Since no data are available in African countries, the prevalence of qnr genes at the University Hospital Ibn Rochd, Casablanca, Morocco, was investigated. Between October 2006 and March 2007, the following 39 randomly selected non-duplicate Enterobacteriaceae producing an extended-spectrum beta-lactamase (ESBL), representing 20% of ESBL strains with respect to species and ward origin, were collected: Escherichia coli (n = 16); Klebsiella spp (n = 14); Enterobacter cloacae (n = 8); Proteus mirabilis (n = 1). Antibiotic susceptibility testing was performed according to CLSI guidelines. ESBL detection was performed by the double disc diffusion test. A multiplex PCR was conducted to detect qnrA, qnrB and qnrS genes that were confirmed by sequencing of the PCR product. The estimated overall prevalence of qnr reached 36% (n = 14; qnrA, 10.25%; qnrB, 23.07%; qnrS, 2.56%). Genes were identified in E. coli, Klebsiella and Enterobacter with a respective prevalence of 18.7%, 50% and 62.5%. The qnr genes were detected in nine wards and qnrA1, qnrB1-B2-B4 and qnrS1 variants were identified. Three genes were identified among nalidixic acid susceptible strains (n = 6); three of those were also susceptible to ciprofloxacin. Among nalidixic acid and ciprofloxacin resistant strains, all strains had qnrB. This study highlights the high prevalence of qnr genes among ESBL strains in the Ibn Rochd CHU, Casablanca. Moreover, qnr were present in quinolone-susceptible strains which could lead to in vivo selection of ciprofloxacin-resistant strains.

Increasing spectrum in antimicrobial resistance of Shigella isolates in Bangladesh: resistance to azithromycin and ceftriaxone and decreased susceptibility to ciprofloxacin.

Antimicrobial resistance of Shigella isolates in Bangladesh, during 2001-2002, was studied and compared with that of 1991-1992 to identify the changes in resistance patterns and trends. A significant increase in resistance to trimethoprim-sulphamethoxazole (from 52% to 72%, p < 0.01) and nalidixic acid (from 19% to 51%, p < 0.01) was detected. High, but unchanged, resistance to tetracycline, ampicillin, and chloramphenicol, low resistance to mecillinam (resistance 3%, intermediate 3%), and to emergence of resistance to azithromycin (resistance 16%, intermediate 62%) and ceftriaxone/cefixime (2%) were detected in 2001-2002. Of 266 recent isolates, 63% were resistant to > or =3 anti-Shigella drugs (multidrug-resistant [MDR]) compared to 52% of 369 strains (p < 0.007) in 1991-1992. Of 154 isolates tested by E-test in 2001-2002, 71% were nalidixic acid-resistant (minimum inhibitory concentration [MIC] > or =32 microg/mL) and had 10-fold higher MIC90 (0.25 microg/mL) to ciprofloxacin than that of nalidixic acid-susceptible strains exhibiting decreased ciprofloxacin susceptibility, which were detected as ciprofloxacin-susceptible and nalidixic acid-resistant by the disc-diffusion method. These strains were frequently associated with MDR traits. High modal MICs were observed to azithromycin (MIC 6 microg/mL) and nalidixic acid (MIC 128 micdrog/mL) and low to ceftriaxone (MIC 0.023 microg/mL). Conjugative R-plasmids-encoded extended-spectrum beta-lactamase was responsible for resistance to ceftriaxone/cefixime. The growing antimicrobial resistance of Shigella is worrying and mandates monitoring of resistance. Pivmecillinam or ciprofloxacin might be considered for treating shigellosis with caution.

Fluoroquinolone-resistantSalmonellaParatyphi A

Fluoroquinolone-resistant<i>Salmonella</i>Paratyphi A

Characterization of the molecular mechanisms of quinolone resistance in Yersinia enterocolitica O:3 clinical isolates.

The aim of this study was to determine the roles of mutations in the gyrA and parC genes and the overexpression of efflux pump(s) as mechanisms of resistance to quinolones. Forty-five Yersinia enterocolitica O:3 clinical isolates (41 nalidixic acid-resistant, three nalidixic acid-susceptible and one nalidixic acid-resistant strain obtained in vitro) were analysed. All the nalidixic acid-resistant strains showed mutations in the gyrA gene and none in the parC gene. The presence of the inhibitor produced decreases in the MIC values of nalidixic acid by two to six serial dilution steps in 37 of the 41 nalidixic acid-resistant strains. Meanwhile, the MIC value of ciprofloxacin was affected in two strains whose values diminished three serial dilution steps. The nalidixic acid-resistant mutant obtained in vitro was also affected by the inhibitor decreasing the MIC value of nalidixic acid three serial dilutions steps whereas the MICs for the nalidixic acid-susceptible strains were not affected. Our results show that the high level of resistance to nalidixic acid is likely due to an overexpression of an efflux pump plus a mutation in the gyrA gene, whereas decreased susceptibility to ciprofloxacin is only associated with the presence of a mutation in the gyrA gene.

Mutations in gyrA and parC genes in nalidixic acid-resistant Escherichia coli strains from food products, humans and animals.

Mutations in quinolone targets were analysed in 80 unrelated nalidixic acid-resistant (NALR) Escherichia coli strains whose nalidixic acid and ciprofloxacin MICs ranged from 32 to >256 mg/L and 0.03-64 mg/L, respectively. These strains were isolated from food products (23) and faecal samples from humans (15) and healthy animals (42). Thirteen nalidixic acid-susceptible (NALS) E. coli strains were also analysed. Mutations in gyrA and parC genes were studied by PCR and sequencing. No amino acid changes were detected in GyrA or ParC proteins of the 13 NALS strains. A single change in the GyrA protein was detected in all 61 NALR strains with ciprofloxacin MICs </= 2 mg/L with the following substitutions (number of strains): Ser-83-->Leu (54), Ser-83-->Ala (one), Ser-83-->Val (one), Asp-87-->Asn (two), Asp-87-->Tyr (two) and Asp-87-->Gly (one). A double change in GyrA was found in 18 of 19 NALR strains with ciprofloxacin MICs >/= 4 mg/L. Amino acid substitutions were Ser-83-->Leu, with an additional change [Asp-87-->Asn (15), Asp-87-->Tyr (two) or Asp-87-->His (one)]. The remaining strain (ciprofloxacin MIC 4 mg/L) showed a single Ser-83-->Leu substitution. In respect of the ParC protein, a single change at Ser-80 or Glu-84 was found in 25 of 42 strains, with ciprofloxacin MICs ranging from 0.5 to 32 mg/L. A double substitution (Ser-80-->Ile and Glu-84-->Gly) was found in one strain (ciprofloxacin MIC 64 mg/L). No amino acid changes were detected in the GyrB protein of 18 NALR strains.

High rate of resistance to nalidixic acid in Salmonella enterica: its role as a marker of resistance to fluoroquinolones

High rate of resistance to nalidixic acid in Salmonella enterica: its role as a marker of resistance to fluoroquinolones

Antimicrobial susceptibilities of Campylobacter jejuni and coli by using E-test in Taiwan.

To report the in vitro antibiotic susceptibility of Campylobacter species, we determined the MICs of 6 antibiotics by E-test for 93 human clinical strains and 35 chicken strains. The 6 antimicrobial agents tested were gentamicin, erythromycin, clindamycin, tetracycline, ciprofloxacin, and nalidixic acid. Isolates from humans were significantly more susceptible than chicken strains to erythromycin, clindamycin and ciprofloxacin. Nearly all of the human and chicken strains were susceptible to gentamicin. Among human isolates of C. jejuni, cross-resistance between nalidixic acid and ciprofloxacin was found in 66% of the strains, but none of the nalidixic acid-susceptible strains was resistant to ciprofloxacin. The higher prevalence of ciprofloxacin resistance in this area may be attributable to the large amount use of quinolones in poultry. Because of the high resistance rates of chicken isolates to the commonly used antimicrobial agents, it is necessary to create innovative methods to limit the inappropriate use of antibiotics in poultry in order to prevent the spread of the drug-resistant strains to humans.

Quinolone-resistant Salmonella typhi in Viet Nam: molecular basis of resistance and clinical response to treatment.

Nalidixic acid-resistant Salmonella typhi (NARST) was first isolated in Viet Nam in 1993. Analysis of the quinolone resistance-determining region of gyrA in 20 NARST isolates by polymerase chain reaction and single-stranded conformational polymorphism yielded two novel patterns: pattern II corresponding to a point mutation at nucleotide 87 Asp-->Gly (n = 17), and pattern III corresponding to a point mutation at nucleotide 83 Ser-->Phe (n = 3). In trials of short-course ofloxacin therapy for uncomplicated typhoid, 117 (78%) of 150 patients were infected with multidrug-resistant S. typhi, 18 (15%) of which were NARST. The median time to fever clearance was 156 hours (range, 30-366 hours) for patients infected with NARST and 84 hours (range, 12-378 hours) for those infected with nalidixic acid-susceptible strains (P < .001). Six (33.3%) of 18 NARST infections required retreatment, whereas 1 (0.8%) of 132 infections due to susceptible strains required retreatment (relative risk = 44; 95% confidence interval = 5.6-345; P < .0001). We recommend that short courses of quinolones not be used in patients infected with NARST.

Comparison of disc diffusion and agar dilution methods for antibiotic susceptibility testing of Campylobacter jejuni subsp. jejuni and Campylobacter coli

The correlation between disc diffusion and agar dilution susceptibility testing of five antibiotics was studied against 145 Campylobacter strains: 99 Campylobacter jejuni subsp. jejuni and 46 Campylobacter coli. The percentages of true results and 95% CI for disc diffusion for resistant strains were 100% (93.2-100%) for tetracycline (53 strains tested), 100% (77.2-100%) for ciprofloxacin (13 strains tested), 86.7% (62.1-96.3%) for nalidixic acid (15 strains tested), 100% (56.6-100%) for erythromycin (five strains tested) and 68.8% (44.4-85.8%) for ampicillin (16 strains tested). The percentages of true results and 95% CI were 97.6-100% and 93.2-100% respectively for 89-140 susceptible strains to the five antibiotics tested. There was a 1.4% major error for nalidixic acid, 0.7% very major error for erythromycin, 5.5% and 1.4% minor and major errors respectively for ampicillin. There was complete agreement for ciprofloxacin and tetracycline. Results of ampicillin susceptibility are not expected to be useful in a clinical setting. The nalidixic acid disc is a marker of ciprofloxacin susceptibility as the nalidixic acid-susceptible strains were susceptible to ciprofloxacin while most of the resistant ones were resistant to ciprofloxacin. Overall, our results suggest that disc diffusion is a reliable, easy and inexpensive susceptibility testing method for C. jejuni and C. coli for erythromycin, ciprofloxacin and tetracycline. Until more erythromycin- and ciprofloxacin-resistant strains are tested to confirm the reliability of this test, the resistance to these drugs needs to be confirmed using the Etest or the agar dilution method.

Antibacterial Activities of Grepafloxacin, Ciprofloxacin, Ofloxacin and Fleroxacin

Grepafloxacin (OPC-17116) and three other fluoroquinolones were tested against 461 bacterial isolates representing 44 species. Grepafloxacin was superior to ciprofloxacin, ofloxacin and fleroxacin in its activity against the Gram-positive cocci, including staphylococci, pneumo-cocci and hemolytic streptococci. Attempts to select grepafloxacin-resistant mutants from a methicillin-resistant Staphylococcus aureus strain were unsuccessful, although ciprofloxacin readily selected quinolone-resistant mutants of that strain. Among Gram-negative bacilli other than Escherichia coli, mutants were readily selected by in vitro exposure to either grepafloxacin or ciprofloxacin. All such mutants showed complete cross resistance to six other quinolones. The four study drugs were active against nalidixic acid-susceptible strains belonging to 13 species of the Enterobacteriaceae, but ciprofloxacin was the most potent drug studied. Against Acinetobacter spp. and Stenotrophomonas maltophilia, grepafloxacin was two- to four-times more active than ciprofloxacin. However, against Pseudomonas aeruginosa, ciprofloxacin was two- to four-times more potent than grepafloxacin. Grepafloxacin and ciprofloxacin were both found to be bactericidal agents.

In vitro activities of sparfloxacin, tosufloxacin, ciprofloxacin, and fleroxacin

The in vitro activity of sparfloxacin was compared with those of tosufloxacin, ciprofloxacin, and fleroxacin against 730 bacterial isolates representing 49 different species. Sparfloxacin and ciprofloxacin had similar spectra of activity, but sparfloxacin was less active against Pseudomonas aeruginosa and more active against many gram-positive cocci and anaerobic bacteria. Tosufloxacin MICs were generally 8- to 16-fold lower than those for sparfloxacin or ciprofloxacin. All four fluoroquinolones were active against nalidixic acid-susceptible strains of the family Enterobacteriaceae (MIC for 90% of the isolates [MIC90], less than or equal to 0.25 micrograms/ml) but nalidixic acid-resistant strains were less susceptible (MIC90, greater than or equal to 4.0 micrograms/ml). Against Pseudomonas aeruginosa isolates, MIC90s were 1.0 micrograms/ml for tosufloxacin, 2.0 micrograms/ml for ciprofloxacin, and 4.0 micrograms/ml for sparfloxacin. Against Enterococcus faecalis, sparfloxacin and ciprofloxacin MIC90s were 1.0 and 2.0 micrograms/ml, respectively. MIC90s for ciprofloxacin-susceptible Staphylococcus aureus were 0.016 micrograms/ml for tosufloxacin, 0.06 micrograms/ml for sparfloxacin, and 0.5 micrograms/ml for both ciprofloxacin and fleroxacin. With four species of gram-negative bacilli, mutants resistant to two to four times the sparfloxacin MIC occurred spontaneously at frequencies of 10(-7) to 10(-9): single-step high-level resistance was not observed. In vitro-selected sparfloxacin-resistant mutants displayed cross-resistance to other quinolones, as did clinical isolates of ciprofloxacin-resistant S. aureus. Tosufloxacin MICs with broth microdilution methods were four- to eightfold greater than those obtained with agar dilution methods. The two procedures gave comparable results when sparfloxacin or ciprofloxacin was being tested.

Therapy for shigellosis. I. Randomized, double-blind trial of nalidixic acid in childhood shigellosis

We compared nalidixic acid, 55 mg/kg/day, with ampicillin, 100 mg/kg/day, both given orally for 5 days, in the treatment of children with dysentery caused by shigellosis. All patients entered into the study had illness of less than 72 hours' duration and no prior allopathic drug therapy. Treatment was randomized and administered in double-blind fashion. Patients initially treated with ampicillin who were infected with a Shigella strain resistant to ampicillin were considered as a separate group (ampicillin-R). All isolates were susceptible to nalidixic acid. Similar percentages of patients treated with nalidixic acid (26/32, 81%) and with ampicillin (17/22, 77%) were clinically cured by the end of therapy; the rate in ampicillin-R (3/14, 21%) patients was significantly lower (p less than 0.001). Stool frequency in patients treated with nalidixic acid was significantly less than for ampicillin-treated or ampicillin-R patients during the final 3 study days. All patients treated with nalidixic acid and ampicillin had Shigella eradicated from their stool by day 3, compared with 77% (10/13) of ampicillin-R patients (p less than 0.05, ampicillin-R vs nalidixic acid or ampicillin). We conclude that nalidixic acid is an effective alternative to ampicillin in the treatment of shigellosis caused by nalidixic acid-susceptible strains.

Resistance to ten different fluoroquinolone antibiotics following in vitro exposures to nalidixic acid

Ten different fluoroquinolone antibiotics were tested against selected Enterobacteriaceae . Against nalidixic acid-resistant clinical isolates, minimal inhibitory concentrations for all 10 drugs were eightfold to 32-fold greater than those against comparable nalidixic acid-susceptible strains. When the latter strains were serially exposed to increasing concentrations of nalidixic acid, susceptibility to all 10 fluoroquinolones decreased as resistance to nalidixic acid and cinoxacin was acquired in vitro.