Fluoroquinolone resistance in penicillin-resistant Streptococcus pneumoniae Clones, Spain.Among 2,882 Streptococcus pneumoniae Streptococcus pneu·mo·ni·ae n. Pneumococcus. Streptococcus pneumoniae Microbiology A pathogenic streptococcus with 90 serotypes associated with pneumonia, bacteremia, meningitis Transmission Person to person Incidence sent to the Spanish Reference Laboratory during 2002, 75 (2.6%) were ciprofloxacin-resistant. Resistance was associated with older patients (3.9% in adults and 7.2% in patients [greater than or equal to] 65 years of age), with isolation from noninvasive sites (4.3% vs. 1.0%), and with penicillin and macrolide resistance. Among 14 low-level resistant (MIC 4-8 [micro]g/mL) strains, 1 had a fluoroquinolone fluoroquinolone /flu·o·ro·quin·o·lone/ (-kwin´o-lon) any of a subgroup of fluorine-substituted quinolones, having a broader spectrum of activity than nalidixic acid. fluor·o·quin·o·lone n. efflux efflux Medtalk That which flows outward phenotype phenotype (fē`nətīp'): see genetics. phenotype All the observable characteristics of an organism, such as shape, size, colour, and behaviour, that result from the interaction of its genotype (total genetic makeup) with , and 13 showed single ParC changes. The 61 high-level ciprofloxacin-resistant (MIC [greater than or equal to] 16 [micro]g/mL) strains showed either two or three changes at ParC, ParE, and GyrA. Resistance was acquired either by point mutation point mutation n. A mutation that involves a single nucleotide and may consist of loss of a nucleotide, substitution of one nucleotide for another, or the insertion of an additional nucleotide. (70 strains) or by recombination recombination, process of "shuffling" of genes by which new combinations can be generated. In recombination through sexual reproduction, the offspring's complete set of genes differs from that of either parent, being rather a combination of genes from both parents. with viridans streptococci Streptococcus (plural, streptococci) A genus of spherical-shaped anaerobic bacteria occurring in pairs or chains. Sydenham's chorea is considered a complication of a streptococcal throat infection. (4 strains) at the topoisomerase topoisomerase an enzyme involved in DNA replication that introduces a single-strand nick in the DNA enabling it to swivel and thereby relieve the accumulated winding strain generated during unwinding of the double helix. II genes. Although 36 pulsed-field gel electrophoresis gel electrophoresis n. Electrophoresis performed in a gel composed of agarose, polyacrylamide, or starch. patterns were observed, 5 international multiresistant clones ([Spain.sup.23F]-1, [Spain.sup.6B]-2, [Spain.sup.9V]-3, [Spain.sup.14]-5 and [Sweden.sup.15A]-25) accounted for 35 (46.7%) of the ciprofloxacin-resistant strains. Continuous surveillance is needed to prevent the dissemination of these clones. ********** Resistance of Streptococcus pneumoniae to multiple antibacterial antibacterial /an·ti·bac·te·ri·al/ (-bak-ter´e-al) destroying or suppressing growth or reproduction of bacteria; also, an agent that does this. an·ti·bac·te·ri·al adj. agents, including [beta]-lactams, macrolides, tetracyclines Tetracyclines Definition Tetracyclines are medicines that kill certain infection-causing microorganisms. Purpose Tetracyclines are called "broad-spectrum" antibiotics, because they can be used to treat a wide variety of , and co-trimoxazole, has emerged worldwide in the 1980s and 1990s (1-3) and has emphasized the need for new therapeutic alternatives, such as newer fluoroquinolones. Older fluoroquinolones, such as ciprofloxacin ciprofloxacin /cip·ro·flox·a·cin/ (sip?ro-flok´sah-sin) a synthetic antibacterial effective against many gram-positive and gram-negative bacteria; used as the hydrochloride salt. cip·ro·flox·a·cin n. and ofloxacin, have been widely used in the last 2 decades, but their activity against gram-positive pathogens is limited. Newer fluoroquinolones, such as levofloxacin, gatifloxacin, moxifloxacin, and gemifloxacin, have enhanced activity against most respiratory pathogens, and some are being more widely used to treat respiratory tract infections Noun 1. respiratory tract infection - any infection of the respiratory tract respiratory infection infection - the pathological state resulting from the invasion of the body by pathogenic microorganisms . Therefore, the emergence of fluoroquinolone-resistant S. pneumoniae strains, although worldwide prevalence is low, is a concern to clinicians who manage respiratory tract infections. A global surveillance study from 1998 to 2000 included 8,882 pneumococci isolated from blood and sputum sputum /spu·tum/ (spu´tum) [L.] expectoration; matter ejected from the trachea, bronchi, and lungs through the mouth. sputum cruen´tum bloody sputum. samples that were collected from centers in 26 countries. The study showed a 1.1% prevalence of ofloxacin-resistant strains, although in some places higher values of nonsusceptible ofloxacin strains were observed: 10.1% in Israel, 14.1% in Japan, and 22.3% in Hong Kong Hong Kong (hŏng kŏng), Mandarin Xianggang, special administrative region of China, formerly a British crown colony (2005 est. pop. 6,899,000), land area 422 sq mi (1,092 sq km), adjacent to Guangdong prov. (l). Ciprofloxacin-resistant strains isolated from patients with community-acquired respiratory tract infections have been reported in Spain (3.0% [4] and 7.1% [5]), Canada (1.7% [6]), and the United States United States, officially United States of America, republic (2005 est. pop. 295,734,000), 3,539,227 sq mi (9,166,598 sq km), North America. The United States is the world's third largest country in population and the fourth largest country in area. (1.4% [7]). Fluoroquinolone resistance is higher among related viridans group streptococci (VGS VGS Videregående Skole (Norwegian school) VGS Virtual Game Station VGS Voodoo Glow Skulls (Ska band) VGS Video Game System VGS Volunteer Gliding School VGS Voltage Gate to Source VGS Velocity Gate Stealer ) isolated from blood (8). Increased fluoroquinolone use correlates with an increase in the prevalence of ciprofloxacin resistance (4,6). Prior fluoroquinolone administration is a risk factor for resistant strain selection, as observed for infections caused by S. pneumoniae (9-12) and VGS (13). A study from our group showed ciprofloxacin-resistant pneumococci emerging in a patient who had received long-term ciprofloxacin therapy to treat persistent bronchiectasis bronchiectasis Abnormal expansion of bronchi in the lungs. It usually results when preexisting lung disease causes bronchial inflammation and obstruction. Bronchial wall fibres degenerate, and bronchi become dilated or paralyzed, preventing removal of secretions, which with Pseudomonas aeruginosa Pseudomonas aeruginosa A normal soil inhabitant and human saprophyte that may contaminate various solutions in a hospital, causing opportunistic infection in weakened Pts Clinical Infective endocarditis in IVDAs, RTIs, UTIs, bacteremia, meningitis, 'malignant' infection (14). Bacterial resistance to quinolones occurs mainly by alteration of their intracellular drug targets, the DNA topoisomerase DNA topoisomerase /DNA topo·isom·er·ase/ (to?po-i-som´er-as) either of two types of isomerase that catalyze the breakage, passage, and rejoining of one or both DNA strands, type I topoisomerases IV (Par[C.sub.2]Par[E.sub.2]) and DNA gyrase DNA gyrase (ji´ras) a type II DNA topoisomerase. (Gyr[A.sub.2]Gyr[B.sub.2]) enzymes. The pneumococcal pneumococcal /pneu·mo·coc·cal/ (-kok´al) pertaining to or caused by pneumococci. parC and parE genes are homologous homologous /ho·mol·o·gous/ (ho-mol´ah-gus) 1. corresponding in structure, position, origin, etc. 2. allogeneic. ho·mol·o·gous adj. 1. to gyrA and gyrB, respectively (15,16). Genetic and biochemical studies have shown that fluoroquinolones target primarily topoisomerase IV Topoisomerase IV is one of two type-II topoisomerases in bacteria, the other being DNA gyrase. Like gyrase, topoisomerase IV is able to pass one double-strand of DNA through another double-strand of DNA, thereby changing the linking number of DNA by two in each enzymatic step. and secondarily DNA gyrase in S. pneumoniae (16-19). Resistance mutations are localized in the quinolone resistance-determining regions (QRDRs) of ParC, ParE, and GyrA. Low-level ciprofloxacin-resistant strains had mutations altering the QRDR QRDR Quinolone Resistance-Determining Regions of one of the two subunits of topoisomerase IV: S79 or D83 of ParC (16,17,20) or D435 of ParE (21). High-level ciprofloxacin-resistant strains had changes affecting both QRDRs of ParC and GyrA (S81, E85) (16,17) or ParE and GyrA (21). Genetic transformation experiments showed that single parC mutations confer low-level ciprofloxacin resistance, and that once the cells have acquired this phenotype, transforming to a higher level of resistance was possible by using DNA DNA: see nucleic acid. DNA or deoxyribonucleic acid One of two types of nucleic acid (the other is RNA); a complex organic compound found in all living cells and many viruses. It is the chemical substance of genes. containing the gyrA QRDR from the high-level ciprofloxacin-resistant strains (16,17). The description of recombinant strains with a mosaic structure in their DNA topoisomerase genes (22-24) has established that fluoroquinolone resistance in S. pneumoniae can be acquired by horizontal gene transfer “HGT” redirects here. For other uses, see HGT (disambiguation). Horizontal gene transfer (HGT), also Lateral gene transfer (LGT), is any process in which an organism transfers genetic material to another cell that is not its offspring. as well as by point mutation. VGS, which share the same mechanisms of resistance (13), are donors in the horizontal transfer to pneumococci (25) and act as a reservoir of fluoroquinolone resistance. To investigate the epidemiology of fluoroquinolone-resistant pneumococci, ascertain the possible dissemination of international clones, and determine the incidence of resistant strains originated by interspecific in·ter·spe·cif·ic adj. Arising or occurring between species. interspecific also interspecies Arising or occurring between species. Adj. 1. horizontal transfer, we characterized all ciprofloxacin-resistant S. pneumoniae strains sent to Spanish Reference Laboratory during 2002. Materials and Methods Bacterial Strains, Serotyping, and Susceptibility Tests susceptibility test Antimicrobial susceptibility test, see there We studied 2,882 S. pneumoniae strains submitted from 78 hospitals nationwide to the Spanish Reference Laboratory during 2002. Of the submitted strains, 1,904 (66.1%) were isolated from adults and 978 (39.1%) from children. The origin of isolates was as follows: 1,453 (50.4%) from blood or other sterile sites, 691 (24.0%) from respiratory tract respiratory tract n. The air passages from the nose to the pulmonary alveoli, including the pharynx, larynx, trachea, and bronchi. Respiratory tract , 231 (8.0%) from eye swabs, 205 (7.1%) from ear swabs, and 302 (10.5%) from other sites. Strains were confirmed as S. pneumoniae by standard methods, and serotypes were determined by a quellung reaction quel·lung reaction n. See Neufeld capsular swelling. . Susceptibility tests, performed initially by the agar-dilution method, selected 85 strains (one isolate per patient) with ciprofloxacin MIC [greater than or equal to] 4 [micro]g/mL. The susceptibility of these 85 strains was repeated twice by microdilution (Sensititre commercial plates) according to according to prep. 1. As stated or indicated by; on the authority of: according to historians. 2. In keeping with: according to instructions. 3. the National Committee for Clinical Laboratory Standards (NCCLS NCCLS National Committee for Clinical Laboratory Standards ) methods (26). S. pneumoniae ATCC ATCC American Type Culture Collection, see there 49619 and strain R6 were used for quality control. Fluoroquinolone efflux was determined (27). Genetic Transformation S. pneumoniae strain R6 was grown in a casein casein (kā`sēn), well-defined group of proteins found in milk, constituting about 80% of the proteins in cow's milk, but only 40% in human milk. hydrolysate-based medium (AGCH AGCH Access Grant Channel (GSM) ) with 0.2% sucrose and used as recipient in transformation experiments (20). Cultures containing 4 x [10.sup.6] CFU/mL were treated with DNA at 0.15 [micro]g/mL for 40 min at 30[degrees]C, then at 37[degrees]C for 90 min before plating on media plates containing 2.5 [micro]g/mL of ciprofloxacin. Colonies were counted after 24 h growth at 37[degrees] C in a 5% C[O.sub.2] atmosphere in AGCH medium with 1% agar. Southern Blot Analysis South·ern blot analysis n. An electrophoretic procedure used to separate and identify DNA sequences. Probes for parC and parE were amplified by polymerase chain reaction polymerase chain reaction (pŏl`ĭmərās') (PCR), laboratory process in which a particular DNA segment from a mixture of DNA chains is rapidly replicated, producing a large, readily analyzed sample of a piece of DNA; the process is (PCR PCR polymerase chain reaction. PCR abbr. polymerase chain reaction Polymerase chain reaction (PCR) ) of the R6 strain with oligonucleotides parCUP and parCDOWN, parEUP and parEDOWN, respectively (25). The ant probe was obtained by amplifying strain 3870 DNA (22) with oligonucleotides antUP and antDOWN (25). All probes were labeled with the Phototope-Star Detection Kit (New England Biolabs New England Biolabs (NEB) produces and supplies reagents for the life science industry. NEB offers a large selection of recombinant and native enzymes for genomic research. It also offers products in the areas related to proteomics and drug discovery. , Beverly, MA). Southern blot Southern blot a technique for detecting specific DNA sequences following agar gel electrophoresis of a set of DNA restriction enzyme digestion fragments. The fragments after electrophoresis are transferred to a nitrocellulose or nylon membrane by applying the membrane to the gel; and hybridization hybridization /hy·brid·iza·tion/ (hi?brid-i-za´shun) 1. crossbreeding; the act or process of producing hybrids. 2. molecular hybridization 3. followed the manufacturer's instructions. Pulsed-field Gel Electrophoresis (PFGE PFGE Pulsed-Field Gel Electrophoresis ) Genomic DNA genomic DNA n. The full complement of DNA contained in the genome of a cell or organism. embedded in agarose agarose more highly purified form of agar with similar uses to agar and widely used in the separation of nucleic acid fragments. plugs was restricted with SmaI, and fragments were separated by PFGE in a CHEF-DRIII apparatus (Bio-Rad, Hercules, CA) (28). PFGE patterns were compared to 14 representative international pneumococcal clones of the Pneumococcal Molecular Epidemiology molecular epidemiology Molecular medicine An evolving field that combines the tools of standard epidemiology–case studies, questionnaires and monitoring of exposure to external factors with the tools of molecular biology–eg, restriction endonucleases, Network (28). Strains with patterns varying by three or fewer bands were considered to represent the same PFGE type (29). PCR Amplifications and DNA Sequence DNA sequence Genetics The precise order of bases–A,T,G,C–in a segment of DNA, gene, chromosome, or an entire genome. See Base pair, Base sequence analysis, Chromosome, Gene, Genome. Determination Oligonucleotides gyrA44 and gyrA170 (13) were used to amplify and sequence the gyrA QRDRs from chromosomal DNA. Amplifications were performed with 0.5 U of Thermus thermophilus Thermus thermophilus is a gram negative eubacterium used in a range of biotechnological applications, including as a model organism for genetic manipulation and systems biology. The bacterium is extremely thermophilic, with an optimal growth temperature of about 65ºC. thermostable ther·mo·sta·ble or ther·mo·sta·bile adj. Unaffected by relatively high temperatures, as certain ferments or toxins. DNA polymerase DNA polymerase /DNA po·lym·er·ase/ (pah-lim´er-as) any of various enzymes catalyzing the template-directed incorporation of deoxyribonucleotides into a DNA chain, particularly one using a DNA template. (Biotools, Madrid, Spain), 0.1 [micro]g of chromosomal DNA, 1 [micro]mol/L (each) of the synthetic oligonucleotide Oligonucleotide A deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) sequence composed of two or more covalently linked nucleotides. Oligonucleotides are classified as deoxyribooligonucleotides or ribooligonucleotides. primers, and 0.2 mmol/L of each deoxynucleoside triphosphate triphosphate /tri·phos·phate/ (tri-fos´fat) a salt containing three phosphate radicals. tri·phos·phate n. A salt or ester containing three phosphate groups. (dNTP) in the buffer recommended by the manufacturers. Amplification was achieved with an initial cycle of 1 min denaturation denaturation, term used to describe the loss of native, higher-order structure of protein molecules in solution. Most globular proteins exhibit complicated three-dimensional folding described as secondary, tertiary, and quarternary structures. at 94[degrees]C; 25 cycles of 30 s at 94[degrees]C, 45 s at 55[degrees]C, and 90 s polymerase extension step at 72[degrees]C; and a final 3-min 72[degrees]C extension step, followed by slow cooling to 4[degrees]C. Fragments including ParE residues 398-647, the intergenic parE-parC region, and ParC residues 1-152 were amplified from genomic DNA with oligonucleotides parE398 (13) and parC152 (16) in PCR reactions performed as previously described, except that 30 cycles of amplification with an extension step of 3 min were applied. All strains, except three, CipR-73, CipR-74, and CipR-75, which yielded fragments of 2.4, 3.0, and 2.9 kb, respectively, yielded 1.6-kb fragments. Those PCR fragments that contained both parE and parC QRDRs were purified by using MicroSpin S400 HR columns (Amersham Pharmacia Biotech, Pistcatway, NJ) and sequenced on both strands with oligonucleotides parE398, parE483 (13), parC50, and parC152 (16) with an Applied Biosystems Applied Biosystems, Inc. (formerly NASDAQ: ABIO) is the original name of a pioneer biotechnology company founded in 1981 in Foster City, California, among the Silicon Valley cities of the southern San Francisco Bay Area. Prism 377 DNA sequencer A DNA sequencer is an instrument used to automate the DNA sequencing process. DNA sequencers have become more important due to large genomics projects and the need to increase productivity. , according to protocols provided by the manufacturer. Statistical Analysis The Fisher exact test ([chi square chi square (kī), n a nonparametric statistic used with discrete data in the form of frequency count (nominal data) or percentages or proportions that can be reduced to frequencies. ] test) was used when appropriate. Comparisons were considered significant at p < 0.05. Results Among 2,882 strains, 75 (2.6%) had ciprofloxacin MIC [greater than or equal to] 4 [micro]g/mL by microdilution and were considered ciprofloxacin-resistant. Ten strains with initial ciprofloxacin MIC 4 [micro]g/mL by agar-dilution showed MIC 1-2 [micro]g/mL by microdilution. Among the 75 ciprofloxacin-resistant strains, 67% were isolated from sputum, whereas 19% were from blood, 13% from lower respiratory tract Noun 1. lower respiratory tract - the bronchi and lungs lung - either of two saclike respiratory organs in the chest of vertebrates; serves to remove carbon dioxide and provide oxygen to the blood samples, and 1.3% from pus pus, thick white or yellowish fluid that forms in areas of infection such as wounds and abscesses. It is constituted of decomposed body tissue, bacteria (or other micro-organisms that cause the infection), and certain white blood cells. . Although ciprofloxacin-resistant strains belonged to 17 different serotypes, 6 serotypes accounted for 55 (73.3%) of the strains: 14 (14 strains), 23F (11 strains), 19F (11 strains), 6B (8 strains), 3 (6 strains), and 15A (5 strains). These serotypes were also the most frequent among the 2,807 ciprofloxacin-susceptible strains. No ciprofloxacin-resistance was found among isolates from pediatric patients <15 years, whereas the prevalence of resistance was 75 (3.9%) of 1,904 isolates from adult patients ([greater than or equal to] 15 years), and 53 (7.2%) of 738 isolates from patients [greater than or equal to] 65 years (p < 0.001). The frequencies of ciprofloxacin resistance were higher among noninvasive pneumococci than among invasive strains (61 [4.3%] of 1,429 vs. 14 [1.0%] of 1,453, p < 0.001). An association between penicillin and ciprofloxacin resistance was observed, with ciprofloxacin-resistant strains distributed as follows: 17 (1.0%) of 1,658 penicillin-susceptible, 39 (4.1%) of 961 intermediate-resistant, and 19 (7.2%) of 263 resistant isolates (p < 0.001). Ciprofloxacin-resistant pneumococci were also more represented among macrolide-resistant isolates: 22 (1.2%) ciprofloxacin-resistant strains among 1,833 erythromycin-susceptible and 53 (5.1%) ciprofloxacin-resistant strains among 1,049 erythronycin-resistant (p < 0.001). Antimicrobial-drug resistance among the 2,807 ciprofloxacin-susceptible pneumococci was lower than those of ciprofloxacin-resistant strains (penicillin 41.4% vs. 73.3%, erythromycin erythromycin (ĭrĭth'rōmī`sĭn), any of several related antibiotic drugs produced by bacteria of the genus Streptomyces (see antibiotic). 35.5% vs. 70.7%, tetracycline tetracycline (tĕ'trəsī`klēn), any of a group of antibiotics produced by bacteria of the genus Streptomyces. They are effective against a wide range of Gram positive and Gram negative bacteria, interfering with protein 33.6% vs. 69.3%, and chloramphenicol chloramphenicol (klōr'ămfĕn`əkŏl'), antibiotic effective against a wide range of gram-negative and gram-positive bacteria (see Gram's stain). It was originally isolated from a species of Streptomyces bacteria. 15.4% vs. 44.0%, p < 0.001). Fifty-five (73.3%) of ciprofloxacin-resistant strains showed multidrug resistance multidrug resistance, n the adaptation of tumor cells or infectious agents to resist chemotherapeutic agents. (resistant to more than three chemically unrelated drugs), among them, 26 strains were resistant to six antimicrobial agents Antimicrobial agents Chemical compounds biosynthetically or synthetically produced which either destroy or usefully suppress the growth or metabolism of a variety of microscopic or submicroscopic forms of life. (penicillin, tetracycline, chloramphenicol, erythromycin, clindamycin, and co-trimoxazole). Fourteen (18.7%) ciprofloxacin-resistant (MIC 4-8 [micro]g/mL) strains were classified as low-level ciprofloxacin-resistant, and 61 (81.3%) strains were classified as high-level ciprofloxacin-resistant (MIC 16-128 [micro]g/mL). The comparative in vitro in vitro /in vi·tro/ (in ve´tro) [L.] within a glass; observable in a test tube; in an artificial environment. in vi·tro adj. In an artificial environment outside a living organism. activity of five fluoroquinolones (MI[C.sub.90]) against the ciprofloxacin-resistant strains was the following: gemifloxacin (0.5 [micro]g/mL) > moxifloxacin (4 [micro]g/mL) > gatifloxacin (8 [micro]g/mL) > levofloxacin (32 [micro]g/mL) > ciprofloxacin (64 [micro]g/mL) (Table 1). Gemifloxacin was the most active fluoroquinolone tested by using the NCCLS breakpoint The location in a program used to temporarily halt the program for testing and debugging. Lines of code in a source program are marked for breakpoints. When those instructions are about to be executed, the program stops, allowing the programmer to examine the status of the program (26), 49.3% of ciprofloxacin-resistant strains were nonsusceptible to this antimicrobial antimicrobial /an·ti·mi·cro·bi·al/ (-mi-kro´be-al) 1. killing microorganisms or suppressing their multiplication or growth. 2. an agent with such effects. drug. Among the 14 low-level ciprofloxacin-resistant isolates, 12 were susceptible to levofloxacin, 13 to gatifloxacin, and all were susceptible to moxifloxacin and gemifloxacin according to NCCLS criteria (26). However, all but one of these low-level ciprofloxacin-resistant strains had first-step parC mutations that would favor the appearance of high-level resistant strains. Among the 61 high-level ciprofloxacin-resistant strains, all showed cross-resistance to levofloxacin and gatifloxacin and all but one to moxifloxacin. Twenty four (39.3%) of 61 high-level ciprofloxacin-resistant strains had gemifloxacin MIC 0.12 [micro]g/mL, which could be considered susceptible according to NCCLS (26), although they showed double mutations (parC or parE and gyrA). One of 75 ciprofloxacin-resistant strains, strain CipR-71, with MIC 8 [micro]g/mL, had an efflux mechanism as the single cause of resistance (see below). The parC, parE, and gyrA QRDRs of 85 strains (75 ciprofloxacin-resistant strains and 10 strains with ciprofloxacin MIC 1-2 [micro]g/mL) were characterized. Most resistant strains (70 of 75) showed low nucleotide sequence variations ([less than or equal to] 1%), but five strains had high variations (>4%) in at least one of their QRDRs (Figure). These strains would have a mosaic structure in those genes showing such a QRDR sequence variation (25): two strains in parC, one in parC + parE, and two in parC + parE + gyrA. Mosaic parC genes had the N91D change, and mosaic gyrA genes had the S114G GyrA change (Figure) that is also present in other ciprofloxacin-resistant S. pneumoniae strains with mosaic genes and in both ciprofloxacin-susceptible and -resistant VGS (25), which indicates their recombinational origin. As expected (25), strains with mosaic parE + parC genes had also an ant-like gene in the intergenic parE-parC region (Figure). [FIGURE OMITTED] All low-level ciprofloxacin-resistant strains had mutations producing amino acid amino acid (əmē`nō), any one of a class of simple organic compounds containing carbon, hydrogen, oxygen, nitrogen, and in certain cases sulfur. These compounds are the building blocks of proteins. changes in ParC, whereas 92% of high-level resistant strains had double (ParC + GyrA or ParE + GyrA) and 8% had triple mutations (two changes in ParC + GyrA, ParC + ParE + GyrA, ParC + two changes in GyrA). Classical mutations known to be involved in fluoroquinolone resistance were found in all but two resistant strains with mosaic parC genes (Table 2): D78A (strain CipR-71) and $79R (strain CipR-75). Genetic transformation experiments showed that the $79R change was involved in ciprofloxacin resistance and that the D78A change was not. CipR-71 and CipR-75 chromosomal DNA transformed the susceptible R6 strain to resistance with high efficiency (>1 x [10.sup.4] transformants per microgram microgram /mi·cro·gram/ (µg) (mi´kro-gram) one millionth (10-6) of a gram. mi·cro·gram n. Abbr. of DNA). The analysis of two transformants of each experiment showed that, while transformants obtained with CipR-71 DNA (ParC D78A) did not carry parC mutations, those obtained with CipR-75 (ParC S79R) carried the same nucleotide changes in parC as the donor strain. When PCR products carrying ParE residues 398-647, the intergenic pare--parC region, and ParC residues 1-152 (including ParE and ParC QRDRs) were used as DNA donors, only those obtained from strain CipR-75 were able to transform R6 to ciprofloxacin resistance. The two transformants selected carried the ParC S79R change, and one of them also had the N91D change. These results showed that the S79R change is involved in the resistance phenotype of strain CipR-75 and that the D78A change of strain CipR-71 is not involved in resistance. Determining MIC to ciprofloxacin, norfloxacin, ethidium bromide Ethidium bromide (sometimes abbreviated as EtBr) is an intercalating agent commonly used as a nucleic acid stain in molecular biology laboratories for techniques such as agarose gel electrophoresis. , and acriflavine ac·ri·fla·vine n. A brown or orange powder derived from acridine and used as a topical antiseptic. acriflavine an antiseptic dye used for topical application; average strength is 1:1000 to 1:8000 solution. in the presence or absence of reserpine reserpine (rĕsûr`pēn), alkaloid isolated from the root of the snakeroot plant (Rauwolfia serpentina), a small evergreen climbing shrub of the dogbane family native to the Indian subcontinent. showed greater than fourfold fourfold Adjective 1. having four times as many or as much 2. composed of four parts Adverb by four times as many or as much Adj. 1. MIC decreases in the presence of reserpine, both in the CipR-71 strain and in the two transformants obtained when chromosomal DNA was used as donor, which indicates the existence of an efflux mechanism (data not shown). Although 36 different PFGE patterns were observed in the 75 ciprofloxacin-resistant strains, 48 strains can be grouped into nine PFGE patterns (Table 3): 11 strains belonged to [Spain.sup.23F]-1 clone, 8 strains to [Spain.sup.14]-5 clone, 6 strains to [Spain.sup.9V]-3 clone, 5 strains to [Spain.sup.6B]-2 clone, 5 strains to [Sweden.sup.15A]-25 clone, 4 strains to clone C of serotype serotype /se·ro·type/ (ser´o-tip) the type of a microorganism determined by its constituent antigens; a taxonomic subdivision based thereon. se·ro·type n. See serovar. v. 19F, 4 strains to clone D of serotype 19F, 3 strains to clone A of serotype 3 related to multilocus sequence type (MLST MLST Multi Locus Sequence Typing MLST Medical Logistics Support Team MLST Mini Losi Super Truck (1/18th scale radio control vehicle) ) 260 (30), and 2 strains to clone B of serotype 3 related to MLST 180 (30). Among 14 blood isolates, 12 different PFGE types were observed. Only three of these strains (CipR-1, -2, and -15) shared an identical PFGE (clone A of serotype 3), although they had different parE polymorphisms and different resistance patterns (Table 2). One of the strains with a mosaic parC gene belonged to the [Spain.sup.6B]-2 clone, and one strain with mosaic parC and parE genes belonged to the [Spain.sup.23F]-1 clone. Capsular cap·su·lar adj. Of, relating to, or resembling a capsule. Adj. 1. capsular - resembling a capsule; "the capsular ligament is a sac surrounding the articular cavity of a freely movable joint and attached to the bones" switching was observed in three strains of serotype 14 with the [Spain.sup.9V]-3 clone, one strain of serotype 19F with [Spain.sup.9V]-3 clone, and two strains of serotype 19A with [Spain.sup.23]-1 clone. In general, strains that shared the same PFGE pattern shared identical polymorphisms on their DNA topoisomerase QRDRs with respect to the sequence of the R6 strain (Table 3). For instance, all strains of the [Spain.sup.9V]-3 clone had identical polymorphisms, the same found in the ATCC 700671 strain representative of this clone (14). All but one of the strains belonging to the [Spain.sup.23F]-1 clone had two polymorphisms in ParC (K137N and a change in codon codon: see nucleic acid. G128: GGT GGT ?-glutamyl transferase. GGT Gammaglutamyltransferase, see there instead of GGC GGC Girl Guides of Canada GGC Greenwood Genetic Center (South Carolina) GGC Gwasanaeth Gwaed Cymru (Welsh Blood Service) GGC Generalized Goppa Code GGC Grosvenor Gallery Company ), two polymorphisms in ParE (I460V and a change in codon I476: ATT ATT ammonia tolerance test. instead of ATC ATC Air Traffic Control ATC Average Total Cost ATC Certified Athletic Trainer ATC At the Center (Hartford, Maine retreat center) ATC Applied Technology Council ATC All Things Considered ), and a change in codon Y75 of GyrA (TAT TAT abbr. Thematic Apperception Test TAT 1. tube agglutination test. 2. tetanus antitoxin. TAT instead of TAC 1. TAC - Translator Assembler-Compiler. For Philco 2000. 2. TAC - Terminal Access Controller. ). Identical polymorphisms were found in the ATCC 700669 strain representative of the [Spain.sup.23F]-1 clone (data not shown); the only exception was the [Spain.sup.23F]-1 strain CipR-73 that had parC and parE mosaic genes (Table 3). Other exceptions were three strains of the [Spain.sup.14]-5 clone (CipR-38, CipR-39, and CipR-40) that had an additional GyrA polymorphism polymorphism, of minerals, property of crystallizing in two or more distinct forms. Calcium carbonate is dimorphous (two forms), crystallizing as calcite or aragonite. Titanium dioxide is trimorphous; its three forms are brookite, anatase (or octahedrite), and rutile. (V88I), which were isolated from three patients temporally related in the same hospital, and two other strains, CipR-1 of clone A of serotype 3 and CipR-57 of clone D of serotype 19F that showed different polymorphisms in ParE (Table 3). Discussion The worldwide prevalence of fluoroquinolone resistance in S. pneumoniae is low, although it varies over time, geographic region, age group, and origin of isolates (1,4,6,31). The overall incidence of ciprofloxacin-resistant S. pneumoniae strains in this study was 2.6%, lower than that reported in previous studies (3%-7%) (4,5) conducted in Spain in which noninvasive isolates were predominant. In agreement with those previous studies, noninvasive isolates in this study displayed ciprofloxacin resistance as high as 4.3%. A higher prevalence (7.2%) was seen in patients [greater than or equal to] 65 years, which also agreed with previous reports (4,6), and which possibly reflects increased fluoroquinolone use in this group of patients. The prevalence of ciprofloxacin resistance seen in this study (2.6%) is similar to the 2.5% found among unselected isolates of Bellvitge Hospital (Barcelona) in 2002 (J. Linares, pers. comm.), but it is lower than the 6.3% found in Donostia Hospital (San Sebastian) (E. Perez-Trallero, pers. comm.). A significant relation between resistance to penicillin or macrolides and resistance to ciprofloxacin was observed (Table 1), in agreement with previous reports (4,6). Penicillin- and multidrug-resistant pneumococci have been common in Spain since the 1980s; these strains primarily belong to serogroup 19 and to four multiresistant epidemic clones: [Spain.sup.23F]-1, [Spain.sup.6B]-2, [Spain.sup.9V]-3, and [Spain.sup.14]-5 (30,32,33). The emergence of fluoroquinolone resistance among strains of these clones has been described, (33-36) and in our study, resistance occurred mainly among those four widely disseminated clones (Table 3). Since neither temporal nor geographic relationships were found among most patients infected by these four clones, this finding could reflect the high prevalence of these clones in the Spanish population, as suggested previously (36), rather than the spreading of resistant clones. Exceptions were five strains of the [Spain.sup.14]-5 clone that could be grouped in two temporal clusters. One cluster accounted for two strains (CipR-66 and -67) collected in the same laboratory, with identical polymorphisms and triple mutations in QRDRs. The second cluster accounted for three identical strains of the [Spain.sup.14]-5 clone (CipR-38, -39 and -40), which were recovered from three patients at a laboratory from San Sebastian in a 29-day period. These strains had an additional GyrA polymorphism (V881) not present in other [Spain.sup.14]-5 clone strains and may represent a variant of this clone (33). Nevertheless, the epidemiologic features of these international clones (2,37,38) suggest that dissemination of ciprofloxacin resistance through these isolates is a plausible scenario and may predict a rapid increase of resistance in S. pneumoniae in countries with an increasing use of fluoroquinolones. Also plausible is that recombinant ciprofloxacin-resistant isolates with mosaic DNA topoisomerase genes belonging to the [Spain.sup.23F]-1 and [Spain.sup.6B]-2 international clones (Table 3) have disseminated and that these isolates have acquired resistance mutations by horizontal transfer from VGS. Although a low prevalence (5 [6.7%] of 75) of S. pneumoniae strains with mosaic parC genes among the ciprofloxacin-resistant strains was observed, the existence of these kind of strains (Figure) is an indication that recombination has occurred between resistant VGS and S. pneumoniae, including the prevalent pneumococcal clones. In addition, five resistant strains of the [Sweden.sup.15A]-25 clone, another multiresistant clone previously reported as susceptible (28), were found (Table 3). Although newer fluoroquinolones have enhanced in vitro activity against S. pneumoniae and are less likely to select for resistant isolates, their use to treat respiratory tract infections merits special attention. More than 20 reports of levofloxacin treatment failure have been concurrent with the development of resistance during or after therapy. In some cases, strains susceptible to levofloxacin but with a first-step parC mutation, as a consequence of previous quinolone use, were present before therapy was initiated. Identifying these strains will help avoid therapeutic failures. Since fluoroquinolone resistance is more frequent in the elderly with chronic respiratory diseases who have had long-term quinolone therapy (4,6,10-12,14), recent use of these drugs should contraindicate con·tra·in·di·cate v. To indicate the inadvisability of something, such as a medical treatment. further fluoroquinolone treatment. The use of NCCLS susceptibility breakpoints for newer fluoroquinolones underestimates a high proportion of low-level ciprofloxacin-resistant strains with first-step mutations whose detection should be improved by decreasing those breakpoints as previously suggested (11,39). In the present study, which followed NCCLS criteria (26), 12 (85%) of 14 low-level ciprofloxacin-resistant strains with a single parC mutation were susceptible to levofloxacin (MIC 1-2 [micro]g/mL). These data agree with those of other authors who found parC mutations in 59% of the strains with levofloxacin MIC 2 [micro]g/mL (39). Using a ciprofloxacin resistance breakpoint MIC [greater than or equal to] 4 [micro]g/mL, as was used by Chen et al. (6), would improve detection of these mutant strains. In fact, in our study, no strains with first-step mutations and ciprofloxacin MIC 2 [micro]g/mL were detected, although other authors have found these mutations in up to 29% of this type of strain (39). Although gemifloxacin was the most active quinolone studied, and 39.3% of high-level ciprofloxacin resistance with double mutations were gemifloxacin-susceptible with the NCCLS breakpoint, a patient infected by such strains should not be treated with any quinolone. To avoid treatment failures, using microbiologic breakpoints for quinolone susceptibility would be more prudent than using clinical breakpoints. If fluoroquinolones are widely and indiscriminately used, resistance to fluoroquinolones in S. pneumoniae may become a problem in the near future.
Table 1. In vitro activity of 13 antimicrobial druqs aqainst 75
ciprofloxacin-resistant Streptococcus pneumoniae isolates (a)
MI[C.sub.50] MI[C.sub.90] MIC range
Drug ([micro]g/mL) ([micro]g/mL) ([micro]g/mL)
Penicillin 1 2 0.03-8
Amoxicillin 1 4 0.06-16
Cefotaxime 0.5 1 0.03-8
Erythromycin 128 128 0.06->256
Clindamycin 128 128 0.06->256
Tetracycline 32 64 0.12-64
Chloramphenicol 2 16 2-32
Cotrimoxazole [greater than or [greater than or 0.5/9.5-[greater
equal to] 4/76 equal to] 4/76 than or equal
to] 4/76
Ciprofloxacin (d) 32 64 4-128
Levofloxacin 16 32 1-64
Gatifloxacin 4 8 0.5-16
Moxifloxacin 2 4 0.12-8
Gemifloxacin 0.12 0.5 0.06-2
Susceptible
Drug breakpoints %S %I
Penicillin [less than or 26.7 48.0
equal to] 0.06
Amoxicillin [less than or 80.0 12.0
equal to] 2
Cefotaxime [less than or 92.1 5.3
equal to] 1
[less than or 56.1 36.0
equal to] 0.5
Erythromycin [less than or 29.3 0.0
equal to] 0.25
Clindamycin [less than or 37.3 0.0
equal to] 0.25
Tetracycline [less than or 30.7 2.7
equal to] 2
Chloramphenicol [less than or 56.0
equal to] 4
Cotrimoxazole [less than or 32.0 4.0
equal to]
0.5/9.5
Ciprofloxacin (d) NA NA NA
Levofloxacin [less than or 16.0 2.7
equal to] 2
Gatifloxacin [less than or 17.3 2.7
equal to] 1
Moxifloxacin [less than or 20.0 37.3
equal to] 1
Gemifloxacin [less than or 50.7 28.0
equal to] 0.12
Drug %R %I+R
Penicillin 25.3 73.3
Amoxicillin 8.0 20.0 (b)
Cefotaxime 2.6 7.9 (b)
7.9 43.9 (c)
Erythromycin 70.7 70.7
Clindamycin 62.7 62.7
Tetracycline 66.6 69.3
Chloramphenicol 44.0 44.0
Cotrimoxazole 64.0 68.0
Ciprofloxacin (d) NA
Levofloxacin 81.3 84.0
Gatifloxacin 80.0 82.7
Moxifloxacin 42.7 80.0
Gemifloxacin 21.3 49.3
(a) S, susceptible; I, intermediate; and R, resistant, according to
National Committee for Clinical Laboratory Standards (NCCLS) 2004
interpretative criteria; NA, not available.
(b) NCCLS 2004 nonmeningitis criteria.
(c) NCCLS 2004 meningitis criteria.
(d) Ciprofloxacin resistance defined as [greater than or equal to]
4 [mirco]g/mL by Chen et al. (4).
Table 2. Fluoroquinolone MIC of 85 strains and amino acid changes
in their DNA topoisomerase genes (a)
Amino acid substitution
No.
strains ParC Par E GyrA
10 None None None
1 None# None None
8 S79F None None
3 S79Y None None
1 D83G None None
1 D83Y None None
1 S79A None S81F
21 S79F None S81F
9 S79F None S81Y
6 S79F None E85K
6 S79Y None S81F
1 S79Y None S81Y
2 D83N None S81F
1 D83N None S81F
3 None D435N S81F
1 None D435N S81Y
1 S79Y, D83N None E85K
3 S79F D435N E85K
1 S79Y D435N E85K
1 S79F None S81F, E85A
1 S79I# None S81F
1 S79F# None# S81F
1 S79F# None# S81F#
1 S79R# None# S81F#
MIC ([micro]g/mL)
No.
strains CIP LVX GAT MXF GEMI
10 1-2 1-2 0.12-0.5 0.12-0.25 0.015-0.03
1 8 2 0.5 0.12 0.06
8 4-8 2-4 0.5-2 0.12-0.5 0.03-0.06
3 4-8 2-4 0.5 0.25 0.03-0.06
1 4 1 0.5 0.25 0.03
1 4 1 0.5 0.25 0.03
1 16 8 4 2 0.12
21 32-128 8-32 4-8 2-4 0.12-1
9 32-64 8-16 4-8 1-4 0.12-0.25
6 32-64 16-64 4-16 2-8 0.25-2
6 32-64 8-64 4-8 2-4 0.12-0.25
1 32 16 8 4 0.12
2 32-64 8-16 2-4 2 0.12-0.25
1 32 16 8 4 0.5
3 16-32 8-16 4-8 2-4 0.12-0.25
1 16 16 4 2 0.12
1 64 32 8 4 1
3 64 64 8-16 8 0.5
1 64 32 8 4 0.5
1 64 64 16 8 1
1 32 16 4 2 0.12
1 32 16 4 4 0.5
1 32 8 4 2 0.25
1 32 16 8 4 0.5
Only changes involved in resistance are shown, and double underlining
indicates that the residue is located in a gene with a mosaic
structure. Additional amino acid changes, not involved in resistance,
were: ParC D78A (the CipR-71 strain with no mutations that has a
mosaic parC gene), R36C (3 strains), ParC K137N (27 strains), ParC
N91D (the three strains with mosaic parC genes), ParE I460V
(40 strains), ParE453S (1 strain), GyrA V88I (3 strains), GyrA S114G
(the two strains with mosaic gyrA genes), and GyrA N150H (one of the
two strains with a mosaic gyrA gene). CIP, ciprofloxacin; LVX,
levofloxacin; GAT, gatifloxacin; MXF, moxifloxacin; GEMI,
gemifloxacin.
Note: The residue is located in a gene with a mosaic structure
indicated with #.
Table 3. Summary of phenotypic characteristics and changes
in the QRDR among the most prevalent pulsed-field gel
electrophoresis patterns of ciprofloxacin-resistant strains (a)
PFGE Strain Serotype Resistance pattern
Referent R6 NT S
[Spain.sup. CipR-8 23F PECITCSxTCp
23F] - 1
CipR-5 19A (b) PTCSxTCp
CipR-9 23F PECITCSxTCp
CipR-12 23F PTCCp
CipR-30, -31, 23F PECITCSxTCp
-32, -33
CipR-48, -49 23F PECITCSxTCp
CipR-73 19A (b) ECITSxTCp
[Spain.sup. CipR-24 14 PECITCSxTCp
14] - 5
CipR-41 14 PECITCSxTCp
CipR-55 14 PECITCSxTCp
CipR-66, -67 14 PECITCSxTCp
CipR-38, 14 PECITCSxTCp
-39, -40
[Spain.sup. CipR-10 9V PSxTCp
9V] - 3
CipR-14 14 (b) PESxTCp
CipR-51 14 (b) PSxTCp
CipR-20 9V PSxTCp
CipR-28 19F (b) PSxTCp
CipR-58 14 (b) PSxTCp
[Spain.sup. CipR-3 6B PECITSxTCp
6B] - 2
CipR-17, 6B PECITCSxTCp
-18, -19
CipR-72 6B PECITSxTCp
[Sweden.sup. CipR-4 15A PECITCp
15A] - 25
CipR-64 15A PECITCp
CipR-60 15A PECITCp
CipR-45, -50 15A PECITCp
C CipR-59 19F PTCSxTCp
CipR-43, -44 19F PTCSxTCp
CipR-65 19F PECITCSxTCp
D CipR-57 19F PECITCp
CipR-29 19F PECITCCp
CipR-46, -47 19F PECITCp
A CipR-1 3 Cp
CipR-2 3 Cp
CipR-15 3 TCCp
B CipR-61 3 Cp
CipR-52 3 ECp
aa and nt changes in QRDR
PFGE Strain ParC ParE GyrA
Referent R6 None None None
[Spain.sup. CipR-8 G128 (GGT), 1460V, 1476 Y75 (TAT)
23F] - 1 K137N, S79F* (ATT)
CipR-5 G128 (GGT), 1460V, 1476 Y75 (TAT)
K137N, S79F* (ATT)
CipR-9 G128 (GGT), 1460V, 1476 Y75 (TAT)
K137N, S79Y* (ATT)
CipR-12 G128 (GGT), 1460V, 1476 Y75 (TAT)
K137N, D83G* (ATT)
CipR-30, -31, G128 (GGT), 1460V, 1476 Y75 (TAT),
-32, -33 K137N, S79F* (ATT) S81F*
CipR-48, -49 G128 (GGT), 1460V, 1476 Y75 (TAT),
K137N, S79F* (ATT) E85K*
CipR-73 S79F*# None Y75 (TAT),
S81F*
[Spain.sup. CipR-24 S79F* None Y75 (TAT),
14] - 5 S81F*
CipR-41 S79F* None Y75 (TAT),
S81Y*
CipR-55 S79Y* None Y75 (TAT),
S81F*
CipR-66, -67 S79F* D435N* Y75 (TAT),
E85K*
CipR-38, S79F* None Y75 (TAT),
-39, -40 V881, S81Y*
[Spain.sup. CipR-10 K137N, S79Y* 1460V Y75 (TAT)
9V] - 3
CipR-14 K137N, S79A* 1460V Y75 (TAT),
S81F*
CipR-51 K137N, S79Y* 1460V Y75 (TAT),
S81F*
CipR-20 K137N, S79F* 1460V Y75 (TAT),
S81F*
CipR-28 K137N, S79F* 1460V Y75 (TAT),
S81F*
CipR-58 K137N, D83N* 1460V Y75 (TAT),
S81F*
[Spain.sup. CipR-3 S79F* None Y75 (TAT)
6B] - 2
CipR-17, S79F* None Y75 (TAT),
-18, -19 S81F*
CipR-72 S79F*# None Y75 (TAT),
S81F*
[Sweden.sup. CipR-4 G77 (GGA), 1460V Y75 (TAT)
15A] - 25 S79F*
CipR-64 G77 (GGA) 1460V, D435N* Y75 (TAT),
S81Y*
CipR-60 G77 (GGA), 1460V Y75 (TAT),
D83Y* E85K*
CipR-45, -50 G77 (GGA), 1460V Y75 (TAT),
S79F* E85K*
C CipR-59 D83N* None Y75 (TAT),
S81F*
CipR-43, -44 S79F* None Y75 (TAT),
S81Y*
CipR-65 S79Y, D83N* None Y75 (TAT),
E85K*
D CipR-57 G77 (GGA), 1460V, P454S Y75 (TAT),
S79Y* S81Y*
CipR-29 G77 (GGA), 1460V Y75 (TAT),
S79F* S81F*
CipR-46, -47 G77 (GGA), 1460V Y75 (TAT),
S79F* E85K*
A CipR-1 R95C, S79F* 1460V Y75 (TAT),
H104 (CAC)
CipR-2 R95C, S79F* None Y75 (TAT),
H104 (CAC)
CipR-15 R95C, S79F* None Y75 (TAT),
H104 (CAC),
S81F*
B CipR-61 None 1460V, D435N* Y75 (TAT),
S81F*
CipR-52 S79Y* 1460V Y75 (TAT),
S81F*
(a) QRDR, quinolone-resistance determining regions; PFGE, pulse-field
gel electrophoresis Smal patterns; aa, amino acid; nt, nucleotide; S,
susceptible to all antibiotics tested; P, resistant to penicillin (MIC
0.12-4 [micro]g/mL); T, resistant to tetracycline (MIC [greater than
or equal to] 4 [micro]g/mL); C, resistant to chloramphenicol (MIC
[greater than or equal to] 8 [micro]g/mL); E, resistant to erythromycin
(MIC [greater than or equal to] 0.5 [micro]g/mL); CI, resistant to
clindamycin (MIC [greater than or equal to] 0.5 [micro]g/mL); SxT,
resistant to trimethropin-sulfamethoxazole (MIC [greater than or equal
to] 4/76 [micro]g/mL); Cp, resistant to ciprofloxacin (MIC [greater
than or equal to] 4 [micro]g/mL). Residue changes involved in
fluoroquinolone resistance are showed in boldface, and double
underlining indicates that the residue is located in a gene with a
mosaic structure. A, PFGE type related to serotype 3 with MLST 260; B,
PFGE type related to serotype 3 with MLST 180; NT, non typeable.
(b) Capsular switching.
Note: Residue changes involved in fluoroquinolone
resistance indicated with *.
Note: The residue is located in a gene with a mosaic
structure indicated with #.
Acknowledgments We thank all Spanish microbiologists who sent strains to Laboratorio de Referencia de Neumococos. L.B. was supported by a fellowship from Instituto de Salud Carlos III Carlos III may refer to:
Dr. de la Campa La Campa is an aldea, or small town, in the Honduran Department of Lempira, located about 18 kilometers by dirt road from Gracias, the largest town in the immediate region. is a research scientist at the Centro Nacional de Microbiologia, Instituto de Salud Carlos III in Madrid, Spain. Her research interest focuses on the molecular basis of antimicrobial resistance in bacteria. References (1.) Jacobs MR, Felmingham D, Appelbaum PC, Gruneberg RN, the Alexander Project Group. The Alexander Project 1998-2000: susceptibility of pathogens isolated from community-acquired respiratory tract infection to commonly used antimicrobial agents. J Antimicrob Chemother. 2003;52:229-46. (2.) Munoz R, Coffey TJ, Daniels M, Dowson CG, Laible G, Casal J, et al. Intercontinental spread of a multiresistant clone of serotype 23F Streptococcus pneumoniae. J Infect Dis. 1991;164:302-6. (3.) Fenoll A, Jado I, Vicioso D, Perez A, Casal J. Evolution of Streptococcus pneumoniae serotypes and antibiotic resistance antibiotic resistance, n the ability of certain strains of microorganisms to develop resistance to antibiotics. antibiotic resistance in Spain: update (1990-1996). J Clin Microbiol. 1998;36:3447-54. (4.) Linares J, de la Campa AG, Pallares R. Fluoroquinolone resistance in Streptococcus pneumoniae. N Engl J Med. 1999;341:1546-7. (5.) Perez-Trallero E, Fernadez-Mazarrasa C, Garcia-Rey C, Bouza E, Aguilar L, Garcia de Lomas J, et al. Antimicrobial susceptibilities of 1,684 Streptococcus pneumoniae and 2,039 Streptococcus pyogenes Streptococcus py·og·e·nes n. A bacterium that causes the formation of pus or of fatal septicemias. Streptococcus pyogenes A common bacterium that causes strep throat and can also cause tonsillitis. isolates and their ecological relationships Ecological Relationships result from the fact that organisms in an ecosystem interact with each other, in the natural world, no organism is an autonomous entity isolated from its surroundings. : results of a l-year (1998-1999) multicenter surveillance study in Spain. Antimicrob Agents Chemother. 2001;45:3334-40. (6.) Chen DK, McGeer A, de Azavedo JC, Low DE. Decreased susceptibility of Streptococcus pneumoniae to fluoroquinolones in Canada. N Engl J Med. 1999;341:233-9. (7.) Doern GV, Heilmann KP, Huynh HK, Rhomberg PR, Coffman SL, Brueggemann AB. Antimicrobial resistance among clinical isolates of Streptococcus pneumoniae in the United States during 1999-2000, including a comparison of resistance rates since 1994-1995. Antimicrob Agents Chemother. 2001;45:1721-9. (8.) de Azevedo JCS JCS abbr. Joint Chiefs of Staff JCS (US) n abbr (= Joint Chiefs of Staff) → Stabschefs pl , Trpeski L, Pong-Porter S, Matsumura S, Network TCBS TCBS Tea Club and Barrovian Society (from Tolkien novel) TCBS The Bear Creek School TCBS Trunked Common Base Station , Low DE. In vitro activity of fluoroquinolones against antibiotic-resistant blood culture isolates of viridans group streptococci across Canada Across Canada was an afternoon program that formerly aired on The Weather Network. The segment ran from early 1999 until mid 2002. The show ran from 3:00PM ET until 7:00 PM ET. . Antimicrob Agents Chemother. 1999;43:2299-301. (9.) Perez-Trallero E, Garcia-Arenzana JM, Jimenez JA, Peris A. Therapeutic failure and selection of resistance to quinolones in a case of pneumococcal pneumonia Pneumococcal Pneumonia Definition Pneumococcal pneumonia is a common but serious infection and inflammation of the lungs. It is caused by the bacterium Streptococcus pneumoniae. treated with ciprofloxacin. Eur J Clin Microbiol Infect Dis. 1990;9:905-6. (10.) Urban C, Rahman N, Zhao X, Mariano N, Segal-Maurer SA, Drlica K, et al. Fluoroquinolone-resistant Streptococcus pneumoniae associated with levofloxacin therapy. J Infect Dis. 2001;184:794-8. (11.) Davidson R, Cavalcanti R, Brunton JL, Darrin J, Bast Bast, in Egyptian religion Bast (băst), ancient Egyptian cat goddess. At first a goddess of the home, she later became known as a goddess of war. The center of her cult was at Bubastis. Her name also appears as Ubast. DJ, de Azevedo JCS, et al. Resistance to levofloxacin and failure of treatment of pneumococcal pneumonia. N Engl J Med. 2002;346:747-50. (12.) Perez-Trallero E, Marimon JM, Iglesias L, Larruskain J. Fluoroquinolone and macrolide treatment failure in pneumococcal pneumonia and selection of multidrug-resistant isolates. Emerg Infect Dis. 2003;9:1159-62. (13.) Gonzalez I, Georgiou M, Alcaide al·cai·de also al·cay·de n. The commander or governor of a fortress in Spain or Portugal. [Spanish, from Arabic al-q F, Balas D, Linares J, de la Campa AG. Fluoroquinolone resistance mutations in the parC, parE, and gyrA genes of clinical isolates of viridans group streptococci. Antimicrob Agents Chemother. 1998;42:2792-8. (14.) de la Campa AG, Ferrandiz MJ, Tubau F, Pallares R, Manresa F, Linares J. Genetic characterization of fluoroquinolone-resistant Streptococcus pneumoniae strains isolated during ciprofloxacin therapy from a patient with bronchiectasis. Antimicrob Agents Chemother. 2003;47:1419-22. (15.) Balas D, Fernandez-Moreira E, de la Campa AG. Molecular characterization of the gene encoding the DNA gyrase A subunit sub·u·nit n. A subdivision of a larger unit. Noun 1. subunit - a monetary unit that is valued at a fraction (usually one hundredth) of the basic monetary unit fractional monetary unit of Streptococcus pneumoniae. J Bacteriol. 1998;180:2854-61. (16.) Munoz R, de la Campa AG. ParC subunit of DNA topoisomerase IV of Streptococcus pneumoniae is a primary target of fluoroquinolones and cooperates with DNA gyrase A subunit in forming resistance phenotype. Antimicrob Agents Chemother. 1996;40:2252-7. (17.) Janoir C, Zeller V, Kitzis M-D, Moreau NJ, Gutmann L. High-level fluoroquinolone resistance in Streptococcus pneumoniae requires mutations in parC and gyrA. Antimicrob Agents Chemother. 1996;40:2760-4. (18.) Pan X-S, Ambler J, Mehtar S, Fisher LM. Involvement of topoisomerase IV and DNA gyrase as ciprofloxacin targets in Streptococcus pneumoniae. Antimicrob Agents Chemother. 1996;40:2321-6. (19.) Fernandez-Moreira E, Balas D, Gonzalez I, de la Campa AG. Fluoroquinolones inhibit preferentially Streptococcus pneumoniae DNA topoisomerase IV than DNA gyrase native proteins. Microb Drug Resist. 2000;6:259-67. (20.) Martin-Galiano AJ, de la Campa AG. High-efficiency generation of antibiotic-resistant strains of Streptococcus pneumoniae by PCR and transformation. Antimicrob Agents Chemother. 2003;47:1257-61. (21.) Perichon B, Tankovic J, Courvalin R Characterization of a mutation in the parE gene that confers fluoroquinolone resistance in Streptococcus pneumoniae. Antimicrob Agents Chemother. 1997;41:166-7. (22.) Ferrandiz MJ, Fenoll A, Linares J, de la Campa AG. Horizontal transfer of parC and gvrA in fluoroquinolone-resistant clinical isolates of Streptococcus pneumoniae. Antimicrob Agents Chemother. 2000;44:840-7. (23.) Bast DJ, de Azevedo JCS, Tam TY, Kilburn L, Duncan C, Mandell LA, et al. Interspecies recombination contributes minimally to fluoroquinolone resistance in Streptococcus pneumoniae. Antimicrob Agents Chemother. 2001;45:2631-4. (24.) Yokota S, Sato K, Kuwahara O, Habadera S, Tsukamoto N, Ohuchi H, et al. Fluoroquinolone-resistant Streptococcus pneumoniae occurs frequently in elderly patients in Japan. Antimicrob Agents Chemother. 2002;46:3311-5. (25.) Balsalobre L, Ferrandiz MJ, Linares J, Tubau F, de la Campa AG. Viridans group streptococci are donors in horizontal transfer of topoisomerase IV genes to Streptococcus pneumoniae. Antimicrob Agents Chemother. 2003;47:2072-81. (26.) National Committee for Clinical Laboratory Standards. Performance standards for antimicrobial susceptibility testing. Fourteenth informational supplement. NCCLS document M100-S14. Wayne (PA): The Committee; 2004. (27.) Ferrandiz MJ, Oteo J, Aracil B, Gomez-Garces JL, de la Campa AG. Drug efflux and parC mutations are involved in fluoroquinolone resistance in viridans group streptococci. Antimicrob Agents Chemother. 1999;43:2520-3. (28.) McGee L, McDougal L, Zhou J, Spratt BG, Tenover FC, George R, et al. Nomenclature nomenclature /no·men·cla·ture/ (no´men-kla?cher) a classified system of names, as of anatomical structures, organisms, etc. binomial nomenclature of major antimicrobial-resistant clones of Streptococcus pneumoniae defined by the pneumococcal molecular epidemiology network. J Clin Microbiol. 2001;39:2565-71. (29.) Tenover FC, Arbeit R, Goering RV, Mickelsen PA, Murray BE, Persing DH, et al. Interpreting chromosomal DNA restriction patterns produced by pulse-field gel electrophoresis: criteria for bacterial strain typing. J Clin Microbiol. 1995;33:2233-9. (30.) Enright MC, Fenoll A, Griffiths D, Spratt BG. The three major Spanish clones of penicillin-resistant Streptococcus pneumoniae are the most common clones recovered in recent cases of meningitis in Spain. J Clin Microbiol. 1999;37:3210-6. (31.) Daily P, Gelling L, Rothrock G, Reingold A, Vugia D, Barrett NL, et al. Resistance of Streptococcus pneumoniae to fluoroquinolones--United States, 1995-1999. MMWR MMWR Morbidity & Mortality Weekly Report Epidemiology A news bulletin published by the CDC, which provides epidemiologic data–eg, statistics on the incidence of AIDS, rabies, rubella, STDs and other communicable diseases, causes of mortality–eg, Morb Mortal Wkly Rep. 2001;50:800-4 (32.) Coffey T, Berron S, Daniels M, Garcia-Leoni M, Cercenado E, Bouza E, et al. Multiply antibiotic-resistant Streptococcus pneumoniae antibiotic-resistant Streptococcus pneumoniae Any of a number of strains of S pneumoniae which are resistant to one or more antibiotics. See S pneumoniae. recovered from Spanish hospitals (1988-1994): novel major clones of serotypes 14, 19F and 15F. Microbiol. 1996;142:2747-57. (33.) Perez-Trallero E, Marimon JM, Gonzalez A, Iglesias L. Spain 14-5 international multiresistant Streptococcus pneumoniae clone resistant to fluoroquinolones and other families of antibiotics. J Antimicrob Chemother. 2003;51:715-9. (34.) Ho PL, Yung RWH RWH Rain Water Harvesting RWH Return With Honor RWH Radar Warning & Homing RWH Read and Write Hold Time , Tsang DNC DNC Democratic National Committee DNC Democratic National Convention DNC Do Not Call DNC Delaware North Companies DNC Domain Name Commissioner DNC Direct Numerical Control DNC Do Not Change DNC Does Not Compute DNC Digital Nautical Chart , Que TL, Ho M, Seto WH, et al. Increasing resistance of Streptococcus pneumoniae to fluoroquinolones: results of a Hong Kong multicentre study in 2000. J Antimicrob Chemother. 2001;48:659-65. (35.) McGee L, Goldsmith CE, Klugman KP. Fluoroquinolone resistance among clinical isolates of Streptococcus pneumoniae belonging to international multiresistant clones. J Antimicrob Chemother. 2002;49:173-6. (36.) Alou L, Ramirez M, Garcia-Rey C, Prieto J, de Lencastre H. Streptococcus pneumoniae isolates with reduced susceptibility to ciprofloxacin in Spain: clonal diversity and appearance of ciprofloxacin-resistant epidemic clones. Antimicrob Agents Chemother. 2001;45:2955-7. (37.) Corso A, Severina EP, Petruk VF, Mauriz YR, Tomasz A. Molecular characterization of penicillin-resistant Streptococcus pneumoniae isolates causing respiratory disease in the United States. Microb Drug Resist. 1998;4:325-37. (38.) Shi ZY, Enright MC, Wilkinson P, Griffiths D, Spratt BG. Identification of three major clones of multiply antibiotic-resistant Streptococcus pneumoniae in Taiwanese hospitals by multilocus sequence typing Multilocus sequence typing (MLST) is a technique in molecular biology for the typing of multiple loci. The procedure characterizes isolates of bacterial species using the DNA sequences of internal fragments of multiple (usually seven) housekeeping genes. . J Clin Microbiol. 1998;36:3514-9. (39.) Lim S, Bast D, McGeer A, de Azavedo J, Low DE. Antimicrobial susceptibility breakpoints and first-step parC mutation in Streptococcus pneumoniae: redefining fluoroquinolone resistance. Emerg Infect Dis. 2003;9:833-7. Address for correspondence: Adela G. de la Campa, Unidad de Genetica Bacteriana, Centro Nacional de Microbiologla, Instituto de Salud Carlos III, 28220 Majadahonda, Madrid, Spain; fax: +34-91-509-79-19; email: agcampa@isciii.es Adela G. de la Campa, * Luz Balsalobre, * Carmen Carmen throws over lover for another. [Fr. Lit.: Carmen; Fr. Opera: Bizet, Carmen, Westerman, 189–190] See : Faithlessness Carmen the cards repeatedly spell her death. [Fr. Ardanuy, ([dagger]) Asuncion Fenoll, * Emilio Perez-Trallero,([double dagger double dagger n. A reference mark (  ) used in printing and writing. Also called diesis.Noun 1. ]) Josefina Linares, ([dagger]) and the Spanish Pneumococcal Infection Study Network G03/103 (1) * Instituto de Salud Carlos III, Majadahonda, Madrid, Spain; ([dagger]) Hospital de Bellvitge, L'Hospitalet de Llobregat L'Hospitalet de Llobregat or L'Hospitalet is a city to the immediate southwest of Barcelona, Catalonia, Spain, and the second largest in Catalonia by population. L'Hospitalet de Llobregat is notable for being one of the most densely populated cities not just in Spain but , Barcelona, Spain; and ([double dagger]) Hospital Donostia, San Sebastian, Guipuzcoa, Spain (1) Spanish Pneumococcal Infection Study Network--general coordination: Roman Pallares; participants and centers: Ernesto Garcia (Centro de Investigaciones Biologicas, Madrid); Julio Casal, Asuncion Fenoll, Adela G. de la Campa (Centro Nacional de Microbiologia, Institute de Salud Carlos III, Madrid); Emilio Bouza, (Hospital Gregorio Maranon, Madrid); Fernando Baquero (Hospital Ramon y Cajal Ra·mòn y Ca·jal , Santiago 1852-1934. Spanish histologist. He shared a 1906 Nobel Prize for research on the nervous system. , Madrid); Francisco Soriano, Jose Prieto (Fundacion Jimenez Diaz y Hospital Clinico, Madrid); Roman Pallares, Josefina Linares (Hospital Universitari de Bellvitge, Barcelona); Javier Garau, Javier Martinez Javier Martinez (born February 5, 1977, in Bayamon, Puerto Rico) was a Major League Baseball pitcher. Drafted by the Chicago Cubs in the 3rd round of the 1994 MLB amateur draft, Martinez would make his Major League Baseball debut with the Pittsburgh Pirates on April 2, 1998, Lacasa (Hospital Mutua de Terrassa, Barcelona); Cristina Latorre (Hospital Sant Joan Sant Joan (official name; Spanish: San Juan) is a municipality on Majorca, Spain, situated in the center of the island in the comarca of Pla de Mallorca. The town Sant Joan, formerly known as Sant Joan de Sineu, was founded in 1300. de Deu, Barcelona); Emilio Perez-Trallero, Alberto Gonzalez (Hospital Donostia, San Sebastian); Juan Garcia Juan Garcia can refer to
|
|
||||||||||||||||||||
Printer friendly
Cite/link
Email
Feedback
Reader Opinion