Minimum Inhibitory Concentrations Of Enrofloxacin Research Articles

Prevalence of Mycoplasma bovis in Algeria and Characterisation of the Isolated Clones.

Bovine respiratory disease (BRD) is common in calves in Algeria, but to date, Mycoplasma bovis has never been monitored as a potential etiological agent. Here, to assess the presence (direct detection) and circulation (indirect detection) of M. bovis, broncho-alveolar lavage fluids (BALF) and serum samples were collected from 60 veal calf farms in Algeria. A commercial ELISA kit (ID Screen® ELISA) was used to screen for the presence of specific antibodies against M. bovis in 351 blood sera collected from both diseased and healthy calves, and 69% (241 sera) tested positive. BALFs from the 176 diseased calves were used to screen for M. bovis by real-time-PCR (rt-PCR), and 102 (58%) tested positive. A non-exhaustive set of 53 clones were isolated from 44 calves and further subtyped using polC gene sequencing. No predominant subtype was found, and two clones exhibited a new subtype. Fourteen clones were further characterized by multilocus sequence typing, and results showed a high degree of genetic diversity, with some clones having new alleles and subtypes. The minimum inhibitory concentrations (MICs) of 5 antimicrobials regularly used to treat BRD was determined on 45 clones. Susceptibility profiles showed very broad diversity, confirming the variety of clones actively circulating. We detected clones with high MICs, including increased MICs of enrofloxacin (n = 5). This is the first study to report the presence of M. bovis in Algeria in calves with BRD. This research also finds broad genetic and phenotypic diversity in the actively circulating isolates.

Mechanism of enrofloxacin-induced multidrug resistance in the pathogenic Vibrio harveyi from diseased abalones

Vibrio harveyi infection had caused severe economic losses in aquaculture. A pathogenic V. harveyi strain had been successfully induced to be a multiple-resistant strain by enrofloxacin (EFX), then the mechanism of multidrug resistance was analyzed. It suggested that the minimum inhibitory concentration of EFX increased by 32-folds. Results of the Kirby-Bauer test showed that the inhibitory zone diameter was 25.3 mm for the sensitive strain (labeled as HL-S) and 8.5 mm for the resistant strain (labeled as HL-R). After 20 serial passages, even when the stress of EFX was removed, the resistance persisted. After induction of EFX, HL-R resisted to other fluoroquinolones, it even resisted to furazolidone and streptomycin, although it was sensitive to these antibiotics initially. Its sensitivity to rifampicin and doxycycline also decreased obviously. Results showed that 3522 differentially expressed genes were identified. Expression of the multidrugs efflux resistance-nodulation-cell division was significantly upregulated (164.61-folds) in HL-R. Other key genes connected with drug efflux were also upregulated significantly (p<0.05). Notably, recA encoded for recombination protein was upregulated significantly, lexA was downregulated significantly in HL-R. Research results showed that the efflux system and the save our souls system have played crucial roles during the development of multidrug resistance of V. harveyi.

Characterization of Erysipelothrix rhusiopathiae Isolates from Diseased Pigs in 15 Chinese Provinces from 2012 to 2018.

Erysipelothrix rhusiopathiae can cause erysipelas in animals and erysipeloid in humans. Since its recurrence in 2012, swine erysipelas has caused serious losses within the pig industry in China. The aim of this study was to perform multilocus sequence typing and understand the virulence and antimicrobial susceptibility of E. rhusiopathiae isolates in China. Multilocus sequence typing (MLST) of a total of 120 strains was performed, and as a result, three different sequence types were identified, of which ST48 was the main one. Five isolates of each MLST type were randomly selected to be used to challenge mice. ST48 was associated with a higher virulence. Antimicrobial susceptibility was tested using a microdilution technique and, to analyze the resistance mechanism, six strains were selected for genome sequencing. A comparison of the six genomes indicated the presence of a suspected macrolide resistance gene, namely, Erm(A)-like, in erythromycin-resistant strains, which increased the minimum inhibitory concentration (MIC) of erythromycin against E. coli C600 at least four-fold. In addition, three mutations (gyrA86T-I, gyrA90D-N, and parC81S-I) were observed in the quinolone resistance-determining regions (QRDRs) of gyrA and parC in quinolone-resistant strains. After the gyrA gene with the 86T-I mutation or the parC gene with the 81S-I mutation was transfected into E. coli C600, the MIC of enrofloxacin against this strain increased at least two-fold. Our findings provide a theoretical basis for developing antibacterial drugs and may contribute to the clinical prevention and control of E. rhusiopathiae.

Population pharmacokinetics/pharmacodynamics modelling of enrofloxacin for the three major trout pathogens Aeromonas salmonicida, Flavobacterium psychrophilum and Yersinia ruckeri

Enrofloxacin is a fluoroquinolone antimicrobial agent used in freshwater rainbow trout against the main pathogenic bacteria Aeromonas salmonicida, Yersinia ruckeri and Flavobacterium psychrophilum. However, the current “standard” dose (10 mg/kg/day for 10 days) was based only on some old, rather limited experimental data, and needed to be re-assessed. Thus, a pharmacokinetic-pharmacodynamic (PKPD) approach was used by combining a population PK model with new epidemiological data (Minimum Inhibitory Concentrations (MIC)) of the three bacterial species to determine optimal enrofloxacin doses in rainbow trout.Ninety-six rainbow trout (half diploid, half triploid) were randomly assigned to four different groups and received oral (gavage) and then intravenous administration of enrofloxacin at four different doses (range 5–60 mg/kg). Individual blood samples were taken to develop a population PK model.Enrofloxacin should be considered as a long-acting drug in trout due to the observed long plasma half-life (>100 h), which is therefore inadequate with the “standard” dosage based on daily oral administrations. Moreover, the fish ploidy had an impact on the PK of enrofloxacin with a longer persistence of enrofloxacin in triploid individuals, which raises the question of the withdrawal period to apply. The absolute bioavailability of oral enrofloxacin was estimated at ~88%.For F. psychrophilum, the provisional epidemiological cut-off value (CONRI), calculated according to the NRI method, was equal to 0.03 μg/mL. For A. salmonicida and Y. ruckeri, however, no clear bimodal distribution of MIC could be observed, and therefore no relevant CONRI could be obtained.According to our model, a single oral dose of ~5 mg/kg should provide sufficient exposure to treat the wild-type population of F. psychrophilum for 4 days, while complying with the PKPD breakpoints. Then, a maintenance dose of ~2.5 mg/kg could possibly be re-administered every 4 days. The absence of a CONRI did not allow to predict an optimal dose for the two other bacteria. As more than 70% of A. salmonicida isolates in our data set have an enrofloxacin MIC ≥0.25 μg/mL, it seems that enrofloxacin should not be recommended against this bacterium.The PKPD approach allowed us to refine the dosing regimens in rainbow trout, for a more sustainable approach. These new dosing regimens have yet to be clinically confirmed.

The pathogen Mycoplasma dispar Shows High Minimum Inhibitory Concentrations for Antimicrobials Commonly Used for Bovine Respiratory Disease.

Mycoplasma dispar is an overlooked pathogen often involved in bovine respiratory disease (BRD), which affects cattle around the world. BRD results in lost production and high treatment and prevention costs. Additionally, chronic therapies with multiple antimicrobials may lead to antimicrobial resistance. Data on antimicrobial susceptibility to M. dispar is limited so minimum inhibitory concentrations (MIC) of a range of antimicrobials routinely used in BRD were evaluated using a broth microdilution technique for 41 M. dispar isolates collected in Italy between 2011–2019. While all isolates had low MIC values for florfenicol (<1 μg/mL), many showed high MIC values for erythromycin (MIC90 ≥8 μg/mL). Tilmicosin MIC values were higher (MIC50 = 32 μg/mL) than those for tylosin (MIC50 = 0.25 μg/mL). Seven isolates had high MIC values for lincomycin, tilmicosin and tylosin (≥32 μg/mL). More, alarmingly, results showed more than half the strains had high MICs for enrofloxacin, a member of the fluoroquinolone class considered critically important in human health. A time-dependent progressive drift of enrofloxacin MICs towards high-concentration values was observed, indicative of an on-going selection process among the isolates.

Research Note: Longitudinal monitoring of chicken houses in a commercial layer farm for antimicrobial resistance in Escherichia coli with special reference to plasmid-mediated quinolone resistance

Plasmid-mediated quinolone resistance (PMQR) genes located on conjugative plasmids can be transferred to other bacteria in the absence of antimicrobial selective pressure. To elucidate the prevalence of resistance, including PMQR in an egg-producing commercial layer farm in western Japan where no antimicrobials were used, minimum inhibitory concentrations (MIC) for a total of 375 Escherichia coli isolates obtained from chicken houses in the farm between 2012 and 2017 were determined using the agar dilution methods. Eighty-seven isolates resistant to oxytetracycline (OTC) accounted for 23.0% of the tested isolates, followed by isolates resistant to dihydrostreptomycin (DSM) (18.4%), sulfisoxazole (18.1%), ampicillin (AMP) (14.4%), trimethoprim (TMP) (14.4%), and nalidixic acid (10.1%). The prevalence rate of multidrug-resistant (MDR) isolates—which are resistant to 3 or more antimicrobial classes, including β-lactams, aminoglycosides, quinolones, folate pathway inhibitors, tetracyclines, and phenicols—was inversely related to the age of chickens at the time of bacterial examination. Probably, the prevalence of MDR isolates in layer chickens may have decreased with age owing to the absence of selective pressure. Furthermore, 45 isolates exhibiting enrofloxacin MICs of more than 0.25 μg/mL were examined for PMQR genes. The transfer of PMQR genes was tested by conjugation analysis. Southern blot analysis of genomic DNA revealed that the qnrS1 (5 isolates), qnrS2 (1 isolate), and qnrS13 genes (1 isolate) were located on plasmids with sizes ranging from approximately 60 to 120 kpb. In 1 of the 5 qnrS1-positive isolates and in an isolate with qnrS13, the qnrS genes were transferred to recipient strains. The plasmid harboring the qnrS1 gene was typed as IncF by PCR-based replicon typing. On this plasmid, the blaTEM, aadA, tetA, and dfrA1 genes responsible for resistance to AMP, DSM, OTC, and TMP, respectively, were detected. The tetA gene was detected in the plasmid harboring the qnrS13 gene, which was typed as IncI1. These results suggest that despite the low prevalence of quinolone resistance in this farm, various PMQR genes, located on diverse plasmids, exist.

Pharmacokinetics of enrofloxacin and its metabolite ciprofloxacin following single-dose subcutaneous injection in black-tailed prairie dogs (Cynomys ludovicianus).

OBJECTIVE To determine plasma concentrations of enrofloxacin and its active metabolite ciprofloxacin following single-dose SC administration to black-tailed prairie dogs (Cynomys ludovicianus). ANIMALS 8 captive healthy 6-month-old sexually intact male black-tailed prairie dogs. PROCEDURES Enrofloxacin (20 mg/kg) was administered SC once to 6 prairie dogs and IV once to 2 prairie dogs. A blood sample was collected from each animal immediately before (0 hours) and 0.5, 1, 2, 4, 8, 12, and 24 hours after drug administration to evaluate the pharmacokinetics of enrofloxacin and ciprofloxacin. Plasma enrofloxacin and ciprofloxacin concentrations were quantified with ultraperformance liquid chromatography-mass spectrometry, and noncompartmental pharmacokinetic analysis was performed. RESULTS Enrofloxacin was biotransformed to ciprofloxacin in the prairie dogs used in the study. For total fluoroquinolones (enrofloxacin and ciprofloxacin), the mean (range) of peak plasma concentration, time to maximum plasma concentration, and terminal half-life after SC administration were 4.90 μg/mL (3.44 to 6.08 μg/mL), 1.59 hours (0.5 to 2.00 hours), and 4.63 hours (4.02 to 5.20 hours), respectively. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that administration of enrofloxacin (20 mg/kg, SC, q 24 h) in black-tailed prairie dogs may be appropriate for treatment of infections with bacteria for which the minimum inhibitory concentration of enrofloxacin is ≤ 0.5 μg/mL. However, clinical studies are needed to determine efficacy of such enrofloxacin treatment.

Characteristics of the antimicrobial resistance of Staphylococcus aureus isolated from chicken meat produced by different integrated broiler operations in Korea

Vertical integration of the broiler industry allows producers to combine different biosecurity and sanitation practices, housing technologies, and feeding regimens to improve food safety. The purpose of this study was to investigate the prevalence of Staphylococcus aureus (S. aureus) and to characterize the antimicrobial-resistant isolates recovered from 7 different integrated broiler operation systems in Korea. Among 200 chicken meat samples, 94 were observed to be positive for S. aureus. However, the prevalence varied from 25.0 to 58.3% in chicken meats, indicating variation in S. aureus occurrence among the operations. Four methicillin-resistant S. aureus isolates (MRSA) were recovered from 3 different operations. A high proportion of the S. aureus isolates were resistant to penicillins (51.2%), tetracycline (38.8%), and ciprofloxacin (CIP; 33.9%). Especially, 3 different operations showed a high number of CIP resistance (45.5∼100%) and multidrug resistance (50.0∼100%). Among 41 CIP-resistant S. aureus isolates, 75.6% showed a double amino-acid exchange of both gyrA and parC, with CIP minimum inhibitory concentrations (MIC) of ≥32 μg/mL. Four MRSA isolates showed resistance to 5 or 7 classes of antimicrobial agents, exhibiting oxacillin, CIP, and enrofloxacin MIC ranges of 16 to 128, 32 to 64, and 8 to 128 μg/mL, respectively, and had double mutations of S84L/S80F in gyrA/parC. Our findings suggest that S. aureus with resistance to important antimicrobial compounds can now be found in association with integrated broiler operations, providing the data to support the development of a monitoring and prevention program in integrated operations.

Combination of Antimicrobials and Essential Oils as an Alternative for the Control of Salmonella enterica Multiresistant Strains Related to Foodborne Disease.

Due to the increase in bacterial resistance to antimicrobials (AMBs) commonly used in veterinary and human medicine, the new strategies for controlling zoonoses focus on the study of natural products with demonstrated AMB activity, such as essential oils (EOs). The aim of this study was to evaluate the in vitro effect of the combination of enrofloxacin (ENR), ceftiofur (CEF), and trimethoprim-sulfamethoxazole (SXT) with cinnamon, clove, oregano, and red thyme EOs against multiple drug-resistant strains of Salmonella enterica. The minimum inhibitory concentration (MIC) of each product was determined by microdilution and "Checkerboard" methods and their combined effect was evaluated against 15 strains of S. enterica. The results were interpreted by the calculation of fractional inhibitory concentration (FIC) and their respective indexes (FICI). Significant susceptibility of all strains to the four EOs was observed. The results showed a synergistic effect between EOs and AMBs tested, highlighting the upper percentage of total synergies of the SXT with the four EOs (FICI ≤0.5 in 60% of assays), and the most effective combination being the one of ENR and cinnamon. The MIC of cinnamon was reduced from 1250 to 312.5 μg/mL and the MIC of ENR from 2 to 0.031 μg/mL. There was no antagonism in the tested combinations (AMBs-EOs). Our results support the combined use of EOs and AMBs for the control of multiresistant strains of S. enterica with a reduction of the minimum effective dose of AMBs and their adverse effects.

Molecular characterisation of resistance to fluoroquinolones in Haemophilus parasuis isolated from China

There are many studies on fluoroquinolone (FQ) resistance in the literature, but little is known regarding the molecular characterisation of FQ-resistant Haemophilus parasuis. In this study, a total of 138 H. parasuis isolates were examined, among which 83 (60.1%) were resistant to enrofloxacin (EFX) and 8 (5.8%) were resistant to levofloxacin (LFX) as determined by Etest. Ten point mutations in the quinolone resistance-determining regions of gyrA, gyrB, parC and parE were detected by PCR and DNA sequencing. Interestingly, 100% of the resistant isolates contained mutations at 87D of gyrA, but other mutations occurred less frequently. Furthermore, it was found that there was synergy between 73S→R/I in parC and other point mutations with respect to FQ resistance. To examine the effect of different point mutations on FQ resistance, the minimum inhibitory concentrations of EFX and LFX were determined for strains generated by site-directed mutagenesis, and three point mutations (gyrA 87D→N, parC 73S→R and parE 551T→A) were shown to be involved in FQ resistance.

Mechanisms accounting for fluoroquinolone multidrug resistance Escherichia coli isolated from companion animals

Multidrug resistance (MDR) is associated with fluoroquinolone (FQ) resistance in companion animal Escherichia coli (E. coli). In this study, gyrA, gyrB, parC, and parE quinolone resistance determining regions (QRDR) were sequenced among uropathogenic E. coli isolates with different resistant phenotypes. Also determined were porin, efflux pump and regulatory gene expression based on quantitative real-time reverse transcriptase PCR (qRT-PCR), the impact of efflux pump inhibition (Phe-Arg-β-naphthylamide) and the presence of plasmid-mediated quinolone resistance (PMQR). Using enrofloxacin as the prototypic FQ, we found that (i) the number of mutations in target genes correlate well with minimum inhibitory concentrations (MICs). A single mutation (Ser83Leu) in gyrA increases FQ MIC in susceptible isolates; subsequent mutations result in resistance that increases from low (enrofloxacin MICs 4–16μg/ml) to high level (enrofloxacin MICs≥128μg/ml) with each progressive mutation. (ii) as MIC increase, acrB activity and the number of drug classes contributing to the MDR phenotype increases; (iii) a consistent relationship between regulatory gene expression and MIC could not be identified; and (iv) qnrS and aac(6′)-Ib-cr gene were detected in 14 and 5 ENRR-MDR isolates containing the target mutation, respectively. Of 13 isolates expressing PDR isolates, 10 (77%) were positive for qnrS gene, and 4 (40%) carried both qnrS and aac(6′)-Ib-cr gene. These findings demonstrated that MDR-associated FQ resistance in canine and feline uropathogenic E. coli reflects a combination of point mutations, enhanced efflux pump activities, and PMQR mechanisms. Point mutations in DNA gyrase, however, are necessary to achieve a clinical level of FQ resistance.

Molecular characterisation of quinolone resistance in Escherichia coli from animals in Turkey

The aim of this study were to detect the gyrA, parC and marR mutations and qnr genes (qnrA, qnrB and qnrS) in 120 strains of Escherichia coli isolated from animals....

Phylogenetic grouping, antibiotic resistance profile, fluoroquinolone susceptibility and ST131 status of canine extraintestinal Escherichia coli isolated from submissions to a veterinary diagnostic laboratory 2005-08

Fluoroquinolones (FQs) are a recommended treatment for Escherichia coli infections in companion animals, particularly in cases of resistance to other drug classes. In a retrospective study, 162 canine clinical E. coli isolates, obtained from veterinary diagnostic submissions (January 2005 - June 2008), were analyzed for phylogenetic group and antibiogram phenotype, using nine antimicrobials and enrofloxacin, ciprofloxacin, moxifloxacin and pradofloxacin minimum inhibitory concentrations (MICs), either in the absence or presence of an efflux pump inhibitor. The isolate susceptibility distribution was bimodal; a high proportion (141/162;87%) showed a sensitivity equivalent to wildtype E. coli (enrofloxacin MIC 0.004 - 0.06 μg/mL), while a minority (4/162;2%) showed reduced susceptibility (enrofloxacin MICs of 0.125 - 0.5 μg/mL), and the remainder (17/162;10%) yielding enrofloxacin MICs in the highlevel resistance range of ≥16 μg/mL. All FQ-resistant isolates were also multidrug resistant. The majority of FQsensitive isolates belonged to phylogenetic group B2 (101/162;62%), and the majority of resistant isolates to group D (8/17;47%). A single resistant B2 isolate and three FQ-sensitive isolates were identified as ST131. Efflux pump activity contributed significantly to MICs for all FQs, except for ciprofloxacin, which may be attributable to its higher polarity compared to the other FQs. These findings confirm a low prevalence of FQ resistance in Australian canine E. coli isolates. Detection of a high moxifloxacin: low ciprofloxacin MIC efflux-associated phenotype (102/162;63%) amongst canine strains may indicate previous exposure to moxifloxacin selective pressure, providing more evidence of exchange of E. coli strains between humans and dogs. The presence of sensitive ST131 strains in the isolate collection does suggest, however, that resistant ST131 strains could potentially emerge under both human and veterinary antimicrobial selection pressure, a risk that could be mitigated by using the most active fluoroquinolone (i.e. pradofloxacin in dogs) against wild-type E. coli at mutant prevention concentrations.

Accurate assessment of antibiotic susceptibility and screening resistant strains of a bacterial population by linear gradient plate

The dynamics of a bacterial population exposed to the minimum inhibitory concentration (MIC) of an antibiotic is an important issue in pharmacological research. Therefore, a novel antibiotic susceptibility test is urgently needed that can both precisely determine the MIC and accurately select antibiotic-resistant strains from clinical bacterial populations. For this purpose, we developed a method based on Fick's laws of diffusion using agar plates containing a linear gradient of antibiotic. The gradient plate contained two layers. The bottom layer consisted of 15 mL agar containing the appropriate concentration of enrofloxacin and allowed to harden in the form of a wedge with the plate slanted such that the entire bottom was just covered. The upper layer consisted of 15 mL plain nutrient agar added with the plate held in the horizontal position. After allowing vertical diffusion of the drug from the bottom agar layer for 12 h, the enrofloxacin concentration was diluted in proportion to the ratio of the agar layer thicknesses. The uniform linear concentration gradient was verified by measuring the enrofloxacin concentration on the agar surface. When heavy bacterial suspensions were spread on the agar surface and incubated for more than 12 h, only resistant cells were able to form colonies beyond the boundary of confluent growth of susceptible cells. In this way, the true MIC of enrofloxacin was determined. The MICs obtained using this linear gradient plate were consistent with those obtained using conventional antibiotic susceptibility tests. Discrete colonies were then spread onto a gradient plate with higher antibiotic concentrations; the boundary line increased significantly, and gene mutations conferring resistance were identified. This new method enables the rapid identification of resistant strains in the bacterial population. Use of the linear gradient plate can easily identify the precise MIC and reveal the dynamic differentiation of bacteria near the MIC. This method allows the study of genetic and physiological characteristics of individual strains, and may be useful for early warning of antibiotic resistance that may occur after use of certain antimicrobial agents, and guide clinical treatment.

Fluoroquinolone resistance mechanisms in multidrug-resistant Escherichia coli isolated from extraintestinal infections in dogs

Fluoroquinolone resistance is an emerging problem in companion animal practice. The present study aimed to determine comparative fluoroquinolone minimum inhibitory concentrations (MICs) for enrofloxacin, marbofloxacin and pradofloxacin and identify plasmid-mediated quinolone resistance (PMQR) mechanisms in 41 multidrug-resistant (MDR) Escherichia coli isolates representing three main clonal groups (CGs) cultured from extraintestinal infections in dogs. All isolates were resistant to fluoroquinolones and the PMQR genes qnrA1, qnrB2, qnrS1 and qepA were identified in isolates from each CG. For a subset of 13 representative isolates, fluoroquinolone chromosomal resistance mechanisms were characterized. CG1 isolates had three mutations in the quinolone resistance determining region (QRDR), two in gyrA (Ser TCG-83→Leu TTG and Asp GAC-87→Asn AAC) and one in parC (Ser AGC-80→Ile ATT), whilst CG2 and CG3 isolates also possessed an additional mutation in parC (Glu GAA-84→Gly GGA) which was reflected in higher fluoroquinolone MICs compared to CG1. Organic solvent tolerance was demonstrated in 8 of the 13 isolates, and all 13 isolates demonstrated enhanced efflux on the basis of a 4-fold decrease or greater in the MIC of enrofloxacin when incubated with an efflux pump inhibitor. A mutation in acrR which can cause overexpression of the AcrAB multidrug efflux pump was detected in CG1 strains. These findings indicate that fluoroquinolone resistance in MDR E. coli isolated from extraintestinal infections in dogs is associated with a combination of target mutations in the QRDRs, transferable PMQR mechanisms and enhanced efflux.

Resistance Mechanism Against Fluoroquinolones inMycoplasma hyopneumoniaeField Isolates

The quinolone resistance-determining regions (QRDR) of gyrA, gyrB, parC, and parE of ten Mycoplasma hyopneumoniae field isolates that were either sensitive (5) or resistant (5) to the fluoroquinolones flumequine and enrofloxacin were characterized. In all five resistant isolates, one point mutation (C --> A) in parC was found, resulting in an amino acid change from serine to tyrosine at position 80 (Escherichia coli numbering). For four of these isolates, this was the only mutation found. These isolates had a minimum inhibitory concentration (MIC) of enrofloxacin of 0.5 microg/ml, whereas for sensitive isolates the MIC of enrofloxacin was < or =0.06 microg/ml. One resistant isolate (Mh 20) had an extra mutation (C --> T) in gyrA resulting in an amino acid change from alanine to valine at position 83 (E. coli numbering), leading to a further increase in the MIC of enrofloxacin (>1 microg/ml). No mutations resulting in an amino acid change were detected in the QRDR of the gyrB and parE genes of the selected isolates. This is the first description of the mechanism of stepwise resistance against fluoroquinolones in M. hyopneumoniae.

Comparison of the in Vitro Susceptibility of Rodent Isolates of Pseudomonas Aeruginosa and Pasteurella Pneumotropica to Enrofloxacin

The objectives of this study were to determine and compare the in vitro enrofloxacin susceptibility of 94 Pseudomonas aeruginosa isolates obtained from enrofloxacin-treated and untreated mice and that of 40 Pasteurella pneumotropica strains and also to assess the efficacy and effects of enrofloxacin treatment of laboratory mice. The minimum inhibitory concentrations (MICs) of enrofloxacin against all the Ps. aeruginosa isolates were in the range of 1 to 4 microg/ml, whereas those against all the P. pneumotropica strains were less than 0.5 microg/ml. The mutation frequency in 54% of the Ps. aeruginosa isolates on treatment with enrofloxacin ranged from 10(-6) to 10(-8); however, none of the P. pneumotropica strains could grow on medium containing more than 3 microg/ml enrofloxacin. Comparison of in vitro enrofloxacin susceptibilities suggested that enrofloxacin was effective for eliminating P. pneumotropica but not for eliminating Ps. aeruginosa for which the MIC of enrofloxacin was more than 1 microg/ml. These results indicated that the enrofloxacin susceptibility of P. pneumotropica was higher than that of Ps. aeruginosa, and that the enrofloxacin treatment might not affect the susceptibility of Ps. aeruginosa.

In vitro antimicrobial susceptibility of Mycoplasma mycoides mycoides large colony and Arcanobacterium pyogenes isolated from clinical cases of ulcerative balanitis and vulvitis in Dorper sheep in South Africa

The in vitro activities of enrofloxacin, florfenicol, oxytetracycline and spiramycin were determined against field isolates of Mycoplasma mycoides mycoides large colony (MmmLC) by means of the broth microdilution technique. The minimum inhibitory concentrations (MICs) of these antimicrobial drugs were determined for a representative number of 10 isolates and 1 type strain. The susceptibility of Arcanobacterium pyogenes to enrofloxacin, oxytetracycline and tilmicosin was determined by means of an agar disk diffusion test. The MICs of enrofloxacin, florfenicol, oxytetracycline and spiramycin were within the ranges of 0.125-0.5, 1.0-2.0, 2.0-4.0 and 4.0-8.0 microg/ml, respectively. This study has shown that resistance of MmmLC against enrofloxacin, florfenicol, oxytetracycline and spiramycin was negligible. All the field strains of A. pyogenes that were tested were susceptible to enrofloxacin, oxytetracycline and tilmicosin with mean inhibition zones of 30.6, 42.3 and 35.8 mm, respectively. Although there is lack of data on in vivo efficacy and in vitro MIC or inhibition zone diameter breakpoints of these antimicrobial drugs for MmmLC, the MIC results indicate that these 4 classes of antimicrobial drugs should be effective in the treatment of ulcerative balanitis and vulvitis in sheep in South Africa.

Salmonella gallinarum gyrA mutations associated with fluoroquinolone resistance

Salmonella enterica subsp. enterica serovar Gallinarum (S. gallinarum) is the causative organism of fowl typhoid, and an outbreak of fowl typhoid in Korea was confirmed in 1992. The aim of this study was to investigate possible changes in fluoroquinolone susceptibility among S. gallinarum isolates from 1995 to 2001, and to analyse mutations of the gyrA gene in fluoroquinolone-resistant isolates. Among 258 S. gallinarum isolates tested by the disk diffusion method, isolates from 1995 (n=18) were susceptible to all fluoroquinolones tested, whereas a number of isolates from 2001 (n=46) showed reduced susceptibility to enrofloxacin (6.5%), ciprofloxacin (10.9%), norfloxacin (52.5%) and ofloxacin (82.6%). The minimum inhibitory concentration range of enrofloxacin, ciprofloxacin, norfloxacin, ofloxacin and danofloxacin increased from < or =0.06 approximately 0.25 microg/ml in 1995 to 2 approximately 8 microg/ml in 2001. When amino acid changes in the gyrA were analysed by DNA sequencing, 22.5% and 14.7% among 258 isolates had a mutation at the Ser-83 and Asp-87 codons, respectively, and the prevalence of these mutants increased from 5.6% in 1995 to 89.1% in 2001. These mutants contained a change from Ser to Phe or Tyr at codon 83, or a change from Asp to Gly, Tyr or Asn at codon 87, and showed a range of minimum inhibitory concentrations of enrofloxacin from 0.5 to 8 microg/ml, ciprofloxacin from 0.25 to 4 microg/ml, norfloxacin from 2 to 32 microg/ml, ofloxacin from 0.5 to 4 microg/ml, and danofloxacin from 0.5 to 4 microg/ml. These results suggested an important association between the gyrA mutations and fluoroquinolone resistance of S. gallinarum.

Determination of skin concentrations of enrofloxacin in dogs with pyoderma

Objective To compare serum and skin concentrations of enrofloxacin in dogs with pyoderma with those of clinically normal dogs and to evaluate concentrations in dogs with superficial versus deep pyoderma. Animals 16 clinically normal dogs and 16 dogs with pyoderma. Procedure Enrofloxacin (approx 5 mg/kg of body weight, PO) was administered daily to all dogs. Serum samples and skin biopsy specimens were obtained on day 1 at 3 hours after drug administration and on day 3 immediately before and 3 hours after drug administration. Samples and specimens were assayed by high-performance liquid chromatography. Morphometric analysis was performed on skin biopsy specimens to determine correlation between inflammatory cells and peak tissue enrofloxacin concentration on day 1. Results Morphometric analysis revealed high correlation between dermal inflammatory cell count and drug concentration in dogs with pyoderma. Conclusions At mean dosage of 5 mg/kg once daily, enrofloxacin tissue concentrations were significantly greater in dogs with pyoderma at 3 hours after pill administration. Enrofloxacin tissue concentration on day 3 at 3 hours after pill administration was 12.4 times the 90% minimum inhibitory concentration of enrofloxacin for Staphylococcus intermedius. Clinical Relevance In dogs with pyoderma, therapeutic tissue concentrations of enrofloxacin are reached as early as 3 hours after drug administration. (Am J Vet Res 1998;59:veterinary medicine-veterinary medicine)