Decreased Ciprofloxacin Susceptibility Research Articles

Multidrug-ResistantSalmonella enterica, Democratic Republic of the Congo

Multidrug-Resistant<i>Salmonella enterica</i>, Democratic Republic of the Congo

Antibiotic Susceptibility Pattern and the Indicator of Decreased Ciprofloxacin Susceptibility of Salmonella enterica Serovar Typhi Isolated from Dhulikhel Hospital, Nepal

Monitoring the antibiotic susceptibility pattern of Salmonella enterica serovar Typhi (S. Typhi) is important for efficiently managing cases of typhoid fever. In this study, the antimicrobial susceptibility patterns of 114 S. Typhi isolates, which were collected from a university hospital in Nepal during July 2009-December 2010, were investigated by disc diffusion assays. All of the S. Typhi isolates were sensitive to amoxycillin-clavulanic acid. More than 95% of the isolates were sensitive to chloramphenicol, ceftazidime, ceftriaxone, and cotrimoxazole. In addition, 1.7% of the studied isolates showed multiple drug resistance patterns. Of the 40 S. Typhi isolates, 32 strains (80%) showed nalidixic acid (NA) resistance with decreased susceptibility to ciprofloxacin (CIP). Importantly, we found the simultaneous presence of NA resistance and decreased susceptibility to CIP, suggesting that the resistance to NA is a reliable indicator of decreased CIP susceptibility (sensitivity, 97.5%; specificity, 100.0%). Furthermore, the sequencing of NA-resistant S. Typhi isolates showed a predominant amino acid alteration in the quinolone resistance-determining region (QRDR) of gyrA gene at position 83 from Ser→Phe. Two isolates with resistance to both CIP and NA had a double-mutation (Ser83→Phe and Asp87→Asn) in the QRDR of the gyrA gene, of which one had an additional amino acid mutation (Ser80→Ilu) in the QRDR of the parC gene.

Prevalence and Mechanisms of Quinolone Resistance Among Selected NontyphoidSalmonellaIsolated from Food Animals and Humans in Korea

The aim of this study was to investigate the prevalence and mechanism of quinolone resistance among selected nontyphoid Salmonella (NTS) isolates. A total of 1279 NTS isolated from food animals (n=692) and humans (n=587) between 1995 and 2009 were investigated by serotyping, antimicrobial susceptibility testing, screening for plasmid-mediated quinolone resistance (PMQR) genes qnr, aac(6')-Ib-cr, and qepA and mutations in the quinolone resistance-determining region (QRDR) of gyrA and parC by PCR, and DNA sequencing. Three hundred thirty (47.7%) of 692 animal isolates and 177 (30.2%) of 587 human isolates were resistant to nalidixic acid. Most animal (94.8%, 313/330) and human (99.4%, 176/177) NTS exhibited decreased ciprofloxacin susceptibility (minimum inhibitory concentration [MIC]: 0.125-2 mg/L). None of them carried qnr or qepA gene. However, aac(6')-Ib was identified in six animal isolates, of which four carried aac(6')-Ib-cr gene. Based on antimicrobial resistance profile, year of isolation, MIC for quinolones and fluoroquinolones, and isolation frequency of serotype, 114 animal and 83 human isolates were tested for QRDR mutations. All contained a single mutation within the QRDR of gyrA at either codon 87 or 83, and 41 of them contained an additional mutation in parC. The most prevalent mutation was Asp87-Tyr (n=107), followed by Asp87-Gly (n=28), Asp87-Asn (n=26), Ser83-Tyr (n=22), and Ser83-Phe (n=14). Point mutations in parC were observed outside the QRDR, which included 40 isolates with Thr57-Ser substitution and 1 Salmonella Typhimurium with a novel Glu51-Lys substitution. In conclusion, a point mutation within the QRDR of gyrA was primarily responsible for quinolone resistance and reduced susceptibility to fluoroquinolones in NTS in Korea. To our knowledge, this is the first report of occurrence of aac(6')-Ib-cr gene among NTS in Korea. The spread of NTS carrying aac(6')-Ib-cr is of serious concern and should be carefully monitored.

Prevalence and role of efflux pump activity in ciprofloxacin resistance in clinical isolates of Klebsiella pneumoniae

We investigated the prevalence and role of efflux pump activity and possible drug influx resistance in ciprofloxacin susceptibility amongst 26 distinct clinical isolates of Klebsiella pneumoniae of varying ciprofloxacin susceptibilities and known quinolone resistance-determining region (QRDR) genotypes. Cellular [(14)C]ciprofloxacin accumulation patterns and the amount of cell-associated [(14)C]ciprofloxacin of mid-logarithmic phase cells were determined before and after challenging with the efflux pump inhibitor carbonyl cyanide 3-chlorophenylhydrazone (CCCP). Most isolates (24/26), and all with ciprofloxacin minimum inhibitory concentrations (MICs) >1µg/ml, had efflux activity that could extrude up to 90% of cell-associated [(14)C]ciprofloxacin; none had significant influx resistance. In isolates with no QRDR mutations, efflux alone reduced ciprofloxacin susceptibility. In isolates with QRDR mutations, the efflux activity varied: in one isolate with no efflux activity, the most common fluoroquinolone resistance-causing QRDR mutation did not bring about clinically significant ciprofloxacin resistance; isolates with multiple mutations had high MICs and, usually, high levels of efflux activity. Fluoroquinolone efflux activity is much more common in clinical isolates of K. pneumoniae than previously reported and it can contribute to decreased ciprofloxacin susceptibility.

A multiplex single nucleotide polymorphism typing assay for detecting mutations that result in decreased fluoroquinolone susceptibility in Salmonella enterica serovars Typhi and Paratyphi A

ObjectivesDecreased susceptibility to fluoroquinolones has become a major problem for the successful therapy of human infections caused by Salmonella enterica, especially the life-threatening typhoid and paratyphoid fevers.MethodsBy using Luminex xTAG beads, we developed a rapid, reliable and cost-effective multiplexed genotyping assay for simultaneously detecting 11 mutations in gyrA, gyrB and parE of S. enterica serovars Typhi and Paratyphi A that result in nalidixic acid resistance (NalR) and/or decreased susceptibility to fluoroquinolones.ResultsThis assay yielded unambiguous single nucleotide polymorphism calls on extracted DNA from 292 isolates of Salmonella Typhi (NalR = 223 and NalS = 69) and 106 isolates of Salmonella Paratyphi A (NalR = 24 and NalS = 82). All of the 247 NalR Salmonella Typhi and Salmonella Paratyphi A isolates were found to harbour at least one of the target mutations, with GyrA Phe-83 as the most common one (143/223 for Salmonella Typhi and 18/24 for Salmonella Paratyphi A). We also identified three GyrB mutations in eight NalS Salmonella Typhi isolates (six for GyrB Phe-464, one for GyrB Leu-465 and one for GyrB Asp-466), and mutations GyrB Phe-464 and GyrB Asp-466 seem to be related to the decreased ciprofloxacin susceptibility phenotype in Salmonella Typhi. This assay can also be used directly on boiled single colonies.ConclusionsThe assay presented here would be useful for clinical and reference laboratories to rapidly screen quinolone-resistant isolates of Salmonella Typhi and Salmonella Paratyphi A, and decipher the underlying genetic changes for epidemiological purposes.

Typhoid in Kenya is associated with a dominant multidrug-resistant Salmonella enterica serovar Typhi haplotype that is also widespread in Southeast Asia.

In sub-Saharan Africa, the burden of typhoid fever, caused by Salmonella enterica serovar Typhi, remains largely unknown, in part because of a lack of blood or bone marrow culture facilities. We characterized a total of 323 S. Typhi isolates from outbreaks in Kenya over the period 1988 to 2008 for antimicrobial susceptibilities and phylogenetic relationships using single-nucleotide polymorphism (SNP) analysis. There was a dramatic increase in the number and percentage of multidrug-resistant (MDR) S. Typhi isolates over the study period. Overall, only 54 (16.7%) S. Typhi isolates were fully sensitive, while the majority, 195 (60.4%), were multiply resistant to most commonly available drugs-ampicillin, chloramphenicol, tetracycline, and cotrimoxazole; 74 (22.9%) isolates were resistant to a single antimicrobial, usually ampicillin, cotrimoxazole, or tetracycline. Resistance to these antibiotics was encoded on self-transferrable IncHI1 plasmids of the ST6 sequence type. Of the 94 representative S. Typhi isolates selected for genome-wide haplotype analysis, sensitive isolates fell into several phylogenetically different groups, whereas MDR isolates all belonged to a single haplotype, H58, associated with MDR and decreased ciprofloxacin susceptibility, which is also dominant in many parts of Southeast Asia. Derivatives of the same S. Typhi lineage, H58, are responsible for multidrug resistance in Kenya and parts of Southeast Asia, suggesting intercontinental spread of a single MDR clone. Given the emergence of this aggressive MDR haplotype, careful selection and monitoring of antibiotic usage will be required in Kenya, and potentially other regions of sub-Saharan Africa.

Treatment of Enteric Fever

Enteric fever is systemic illness caused by Salmonella Typhi and Salmonella Paratyphi A, B and C. It is believed to be a readily treatable illness by many clinicians in the developing world where it is endemic; however, with the emergence of drug resistance to fluoroquinolones, treatment is becoming increasingly difficult. While drugs such as cefixime, previously believed to be effective, have been proven otherwise, new agents such as gatifloxacin and azithromycin have proven to be promising. Re-emergence of chloramphenicol sensitive strains in previously resistant areas points towards the concept of antibiotic recycling, preserving the use of older antibiotics. Antibiotic recycling has been used successfully in hospital settings. However, its usefulness in community settings, where the main burden of enteric fever resides, is challenging to manage due to logistics and a lack of infrastructure. Nalidixic acid resistance used to be a marker for clinical response to flouroquinolones; however, recent studies highlight the importance of decreased ciprofloxacin susceptibility as a better marker. Enteric fever, as a public health problem, has been tackled by protection of food and water supplies in the industrialised countries of the world. Nonetheless, that goal seems too far-fetched in the developing world where there are hundreds of villages, towns and cities without adequate infrastructures. Perhaps the key to solving this problem is combining point-of-use-purification of water (by chlorination) with the treatment of illness in the community. Treatment of chronic carriers is also necessary in order to halt the cycles of transmission.

Antimicrobial resistance in typhoidal and nontyphoidal salmonellae

Increasing occurrence of antimicrobial resistance in both typhoidal and nontyphoidal salmonellae is a major public health problem. Recent studies documenting the occurrence and types of resistance, with particular reference to quinolones and extended spectrum cephalosporins, and new approaches to treatment are reviewed. Community and hospital-based studies in different Asian and African countries show widely variable rates of resistance in Salmonella enterica serovars Typhi and Paratyphi A. Occurrence of multidrug resistance has declined in some areas, but the incidence of decreased ciprofloxacin susceptibility has reached high levels, particularly in the Indian subcontinent, and isolates with full resistance to this antimicrobial are increasingly reported. Similar variability in resistance rates occurs among nontyphoidal salmonellae, with variation between serovars and by region. There are reports of plasmid-mediated qnr genes and a variety of extended spectrum cephalosporin resistance genes in nontyphoidal serovars. Two randomized controlled trials report gatifloxacin as a potential treatment option in enteric fever caused by multidrug-resistant isolates with decreased ciprofloxacin susceptibility. Patterns of resistance in Salmonella are constantly changing. Continual surveillance of resistance levels is critical for clinicians to keep abreast of treatment options, but it is often lacking in resource-poor regions of the world with the highest disease burden.

Clinical Response and Outcome of Infection with Salmonella enterica Serotype Typhi with Decreased Susceptibility to Fluoroquinolones: a United States FoodNet Multicenter Retrospective Cohort Study

Patients with typhoid fever due to Salmonella enterica serotype Typhi strains for which fluoroquinolones MICs are elevated yet that are classified as susceptible by the current interpretive criteria of the Clinical and Laboratory Standards Institute may not respond adequately to fluoroquinolone therapy. Patients from seven U.S. states with invasive Salmonella serotype Typhi infection between 1999 and 2002 were enrolled in a multicenter retrospective cohort study. Patients infected with Salmonella serotype Typhi isolates with ciprofloxacin MICs of 0.12 to 1 microg/ml (decreased ciprofloxacin susceptibility but not resistant to ciprofloxacin [DCS]) were compared with patients infected with isolates with ciprofloxacin MICs <0.12 microg/ml for fever clearance time and treatment failure. Of 71 patients, 30 (43%) were female and 24 (34%) were infected with Salmonella serotype Typhi with DCS; the median age was 14 years (range, 1 to 51 years). Twenty-one (88%) of 24 isolates with DCS were resistant to nalidixic acid. The median antimicrobial-related fever clearance times in the DCS and non-DCS groups were 92 h (range, 21 to 373 h) and 72 h (range, 19 to 264 h) (P = 0.010), respectively, and the fluoroquinolone-related fever clearance times in the DCS and non-DCS groups were 90 h (range, 9 to 373 h) and 64 h (range, 34 to 204 h) (P = 0.153), respectively. Four (17%) of 24 patients in the DCS group and 2 (4%) of 46 patients in the non-DCS group (relative risk, 2.5; 95% confidence interval, 1.2 to 5.1) experienced treatment failure. Associations persisted after adjustment for potential confounders. We demonstrate that patients infected with Salmonella serotype Typhi isolates with DCS show evidence of a longer time to fever clearance and more frequent treatment failure. Nalidixic acid screening does not detect all isolates with DCS.

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.

Emergence of multidrug-resistant Salmonella enterica serotype Typhi with decreased ciprofloxacin susceptibility in Bangladesh

During 1989-2002, we studied the antimicrobial resistance of 3928 blood culture isolates of Salmonella enterica serotype Typhi (S. Typhi) in Dhaka, Bangladesh. Overall 32% (1270) of the strains were multidrug-resistant (MDR, resistant to chloramphenicol, ampicillin and trimethoprim-sulphamethoxazole); first detected in 1990 (rate of 8%), increased in 1994 (44%), declined in 1996 (22%, P<0.01 compared to 1994) and re-emerged in 2001 (36%) and 2002 (42%, P<0.01 compared to 1996). An increased MIC of ciprofloxacin (0.25 microg/ml) indicating decreased susceptibility to ciprofloxacin was detected in 24 (18.2%) out of 132 randomly selected strains during 1990-2002; more frequently in MDR than susceptible strains (46.3% vs. 5.5%, P<0.001), and the proportion of them rose to 47% in 2002 from 8% in 2000 (P<0.01). Ciprofloxacin (5 microg) disk diffusion zone diameters of < or =24 mm as break-point had 98% sensitivity and 100% specificity when compared with a ciprofloxacin MIC of 0.25 microg/ml as break-point for decreased susceptibility; being a useful and easy screen test. All strains were susceptible to ceftriaxone. The emergence of MDR S. Typhi with decreased ciprofloxacin susceptibility will further complicate the therapy of typhoid fever because of the lack of optimum treatment guidelines.

Antimicrobial resistance of Salmonella enterica group C strains isolated from humans in Turkey, 2000–2002

Fifty-three Salmonella enterica group C isolates obtained from various human samples (47 stool, 4 blood and 2 urine) in ten provinces of Turkey between 1 July 2000 and 30 June 2002 were serotyped and resistance to antimicrobials was investigated by agar dilution tests. The isolates were identified as S. Choleraesuis (11), S. Hadar (7), S. Irumu (4), S. Virchow (3), S. Tallahassee (3), S. Paratyphi C (2), S. Braenderup (2), S. Othmarschen (2), S. Menston (2), S. Concord (2), S. Infantis (2), S. Kottbus (2), S. Edinburg (1), S. Oranienburg (1), S. Muenchen (1) and S. Malmoe (1). Antimicrobial resistance rates of S. enterica groups C 1 and C 2 were high for ampicillin (26% and 60%, respectively), amoxicillin/clavulanic acid (11% and 40%), chloramphenicol (16% and 27%) and tetracycline (3% and 40%). The percentages of strains sensitive to all antimicrobials were 58% and 33%, respectively. Multiresistance was not observed in group C 1 isolates, but the rate of multiresistant isolates was 13% in group C 2. The rate of decreased ciprofloxacin susceptibility (Cip L) was 61% in serogroup C 1 and 20% in serogroup C 2. These results indicated that S. enterica group C infections in humans were not infrequent in Turkey and that multiple antimicrobial resistance was common within these strains.

Mutations in the gyrA gene in Salmonella enterica clinical isolates with decreased ciprofloxacin susceptibility

The incidence of resistance to nalidixic acid has increased in the University General Hospital of Elche, Spain, and this paper describes the investigation of this phenomenon. This increase was mainly due to an increase of nalidixic-resistant Salmonella enterica serotype Enteritidis. Resistance to nalidixic acid is an indicator of decreased ciprofloxacin susceptibility (sensitivity 100%, specificity 96.7%). Strains that were resistant to nalidixic acid and exhibited decreased susceptibility to ciprofloxacin had a single mutation in QRDR of gyrA: Asp87-Asn, Asp87-Tyr or Ser83-Phe. The sensitivity of S. enterica strains to nalidixic acid should be tested and the breakpoint of ciprofloxacin established by MENSURA applied, instead of that of the NCCLS for these strains.

Predictive value of nalidixic acid resistance for detecting salmonellae with decreased ciprofloxacin susceptibility

Seventy-three Salmonella isolates classified as ciprofloxacin susceptible when using the criteria of the National Committee for Clinical Laboratory Standards were studied for nalidixic acid (NA) resistance. The aim of the study was to determine the predictive value of nalidixic acid resistance in screening for decreased ciprofloxacin susceptibility. We observed that isolates with decreased ciprofloxacin susceptibility were all resistant to nalidixic acid. Identification of nalidixic acid resistance by the disk diffusion method provided 100% sensitivity and a specificity of 98.4% in strains with minimum inhibitory concentrations (MICs) >0.008 mg/l.

Trends in antimicrobial drug resistance in Salmonella enterica serotypes Typhi and Paratyphi A isolated in Europe, 1999–2001

Results of antimicrobial sensitivity tests for strains of Salmonella enterica serotypes Typhi and Paratyphi A isolated from patients in ten European countries between 1999 and 2001 have been transferred electronically to the Enter-net surveillance hub. For Typhi between 22 and 29% of isolates were multiresistant (to four drugs or more) with decreased susceptibility to ciprofloxacin (MIC 0.25–1.0 mg/l) increasing from 20% in 1999 to 26% in 2001. Nineteen of 169 (11%) strains with decreased ciprofloxacin susceptibility were sensitive to nalidixic acid. For Paratyphi A multiple resistance increased from 9% in 1999 to 25% in 2001 and decreased ciprofloxacin susceptibility from 6 to 17%. Clinicians should be aware of the possibility of treatment failures when fluoroquinolones are used as the first-line drug for infections with Typhi and Paratyphi A, particularly for patients recently returning from areas where drug-resistant strains are endemic.

Efficacy and pharmacodynamics of gemifloxacin versus levofloxacin in guinea pig pneumococcal pneumonia induced by strains with decreased ciprofloxacin susceptibility

Quinolone in vivo bactericidal activity was investigated in a guinea pig pneumonia model using three Streptococcus pneumoniae strains with decreasing susceptibility to ciprofloxacin. Treatment regimens resulted in values of AUC 0–24 h and C 30 min similar to those of standard oral regimens in human serum. Efficacy was defined as a significant difference in number of viable bacteria in the lungs compared with the control. Ciprofloxacin, levofloxacin and gemifloxacin were effective against the levofloxacin-susceptible strain. Only gemifloxacin achieved a ≥99.9% reduction versus control against the levofloxacin intermediate-resistant strain. Gemifloxacin achieved a 99.69% reduction and was the only quinolone significantly different from the control ( P<0.05) against the levofloxacin-resistant strain. Gemifloxacin offers in vivo activity against ciprofloxacin- to levofloxacin-resistant pneumococci.

Antibiotic Susceptibilities of Salmonella enterica Serovar Typhi and S. enterica Serovar Paratyphi A Isolated from Patients in Japan

The antibiotic susceptibilities of 62 strains of Salmonella enterica serovar Typhi and 37 strains of S. enterica serovar Paratyphi A were investigated with 18 antibiotics. Eighteen S. enterica serovar Typhi isolates and five S. enterica serovar Paratyphi A isolates were resistant to one or more antimicrobial agents, among which 10 S. enterica serovar Typhi isolates were nalidixic acid resistant and also showed decreased ciprofloxacin susceptibility.