Antimicrobial resistance of Clostridium difficile isolates in a tertiary medical center, Israel

The antimicrobial susceptibilities of 49 Clostridium difficile isolates obtained from patients with C. difficile-associated diarrhea to metronidazole, vancomycin, rifampicin, fusidic acid, doxycycline, and linezolid were determined by the disc diffusion and Etest (Biodisk, Solna, Sweden). Random amplification of polymorphic DNA-PCR amplification assay was performed for studying clonality of isolates. Resistance to metronidazole was found in 2% (1/49 isolates; MIC > or = 256 microg/mL) of isolates and resistance to linezolid in 2% (1/49 isolates; MIC = 24 microg/mL). One isolate showed combined resistance to fusidic acid (by disc diffusion test) and rifampicin (MIC > or = 32 microg/mL). All isolates were sensitive to doxycycline and vancomycin. Molecular typing revealed an absence of clonality among the resistant isolates, whereas the sensitive isolates were monoclonal. Resistance of C. difficile to metronidazole and other antimicrobials including linezolid exists in our institution. This finding should promote exploration of this problem in Israel and clarify the impact of resistance on outcome.     

In vitro activity of ramoplanin against Clostridium difficile, including strains with reduced susceptibility to vancomycin or with resistance to metronidazole

We evaluated the in vitro activity of ramoplanin, an antimicrobial compound that inhibits cell wall synthesis by acting at the level of lipid intermediate formation, against Clostridium difficile. We included strains with reduced susceptibilities to vancomycin (vancomycin-intermediate [Van(i)] strains) or with resistance to metronidazole (Mtz(r)), in order to assess the potential utility of ramoplanin for the treatment of C. difficile-associated diarrhea. We tested the activity of ramoplanin against a total of 105 nonduplicate clinical isolates of toxigenic C. difficile, including 8 Van(i) isolates and 6 Mtz(r) isolates, obtained from our laboratory. Ramoplanin was active against all strains tested at concentrations ranging from 0.03 to 0.5 microg/ml (MICs at which 50 and 90% of isolates were inhibited, 0.25 microg/ml; geometric mean MIC, 0.22 microg/ml). All isolates, independently of their levels of susceptibility to vancomycin or metronidazole, were considered susceptible to ramoplanin (MICs, < or =0.5 microg/ml).     

临床分离艰难梭菌188株的耐药性研究

目的了解艰难梭菌临床分离株对抗菌药物的耐药性及耐药机制。方法以琼脂稀释法测定甲硝唑等12种抗菌药物对2007年3月～2009年3月复旦大学附属华山医院临床分离的艰难梭菌的体外抗菌活性;PCR扩增及DNA测序比对检测耐药基因及耐药决定区突变。结果 188株菌株包括25种核糖型别,以核糖型017最为常见(25%),其次为001与012。未检测到核糖型027,但有1株细菌为核糖型078。所有菌株均对甲硝唑、万古霉素和哌拉西林-他唑巴坦敏感,对莫西沙星、环丙沙星、左氧氟沙星、红霉素、克林霉素、四环素、利福平、利福昔明和夫西地酸的耐药率分别为50.0%、100%、70.7%、77.7%、83.0%、47.9%、19.8%、19.8%和12.8%。主要核糖型菌株耐药性更高。耐药株中耐药基因或耐药决定区突变检出率分别为:ermB基因76%;tetM基因97.8%;gyrA基因突变59.6%;g了rB基因突变11.7%;g了rA与gyrB基因同时突变28.7%;rpoB基因突变100%。仅在2株夫西地酸耐药株中检测到fusA基因突变。结论艰难梭菌对甲硝唑与万古霉素仍呈现敏感,但对氟喹诺酮类药物及红霉素等呈现较高的耐药率。核糖型017等流行株对莫西沙星、利福平等抗菌药物耐药率更高。     

Activity of a novel cyclic lipopeptide, CB-183,315, against resistant Clostridium difficile and other Gram-positive aerobic and anaerobic intestinal pathogens

We evaluated the activity of CB-183,315 against Clostridium difficile, including strains that are resistant to fluoroquinolones and metronidazole and with elevated MICs to vancomycin as well as other Gram-positive intestinal pathogens. The MICs of CB-183,315 against all C. difficile isolates were ≤ 1 μg/ml. CB-183,315 had greater activity than vancomycin and metronidazole against C. difficile isolates and was more active than the comparators against vancomycin-resistant enterococcus (VRE). CB-183,315 also had excellent activity against methicillin-resistant Staphylococcus aureus (MRSA), other Clostridium spp., and Peptostreptococcus spp.     

Antimicrobial susceptibility testing of Clostridium difficile using EUCAST epidemiological cut-off values and disk diffusion correlates

With the emergence of reduced susceptibility of Clostridium difficile to metronidazole and vancomycin the value of antimicrobial susceptibility testing has increased. The aim of our study was to evaluate disk diffusion for susceptibility testing of C. difficile by comparing disk diffusion results with MICs from gradient tests and to propose zone diameter breakpoint correlates for the EUCAST epidemiological cut-off values (ECOFFs) recently published. We tested 211 clinical isolates of C. difficile, from patients with diarrhoea hospitalized at Aarhus and Odense University Hospitals, Denmark. Furthermore, ten clinical isolates of C. difficile from the Anaerobe Reference Laboratory, University Hospital of Wales, with known reduced susceptibility to either metronidazole or vancomycin, were included. Isolates were tested with Etest gradient strips and disk diffusion towards metronidazole, vancomycin and moxifloxacin on Brucella Blood Agar supplemented with hemin and vitamin K. We found an excellent agreement between inhibition zone diameter and MICs. For each MIC value, the inhibition zones varied from 0 to 8 mm, with 93% of values within 6 mm for metronidazole, 95% of values within 4 mm for vancomycin, and 98% of values within 4 mm for moxifloxacin. With proposed zone diameter breakpoints for metronidazole, vancomycin and moxifloxacin of WT ≥ 23 mm, WT ≥ 19 and WT ≥ 20 mm, respectively, we found no very major errors and only major errors below 2%. In conclusion, we suggest that disk diffusion is an option for antimicrobial susceptibility testing of C. difficile.     

Prevalence and association of macrolide-lincosamide-streptogramin B (MLS(B)) resistance with resistance to moxifloxacin in Clostridium difficile

Clostridium difficile remains the leading cause of nosocomially acquired diarrhoea. C. difficile usually exhibits resistance against beta-lactam antibiotics, whereas susceptibility to other drugs may vary. This study investigated the antimicrobial susceptibility of C. difficile to different antibiotics over a period of time and characterizes molecular mechanisms for resistance. One hundred and seventy-three toxigenic and 19 non-toxigenic C. difficile strains, recovered from patients in two university hospitals in Germany between 1986 and 2001, were investigated for their susceptibility to erythromycin, clindamycin, moxifloxacin, vancomycin and metronidazole employing the Etest. The genetic background for resistance was analysed using PCR and DNA sequencing. All strains were susceptible to vancomycin and metronidazole. Resistance to erythromycin, clindamycin and moxifloxacin was found in 27%, 36% and 12% of the tested strains, respectively. High-level resistance (MIC > 128 mg/L) against erythromycin and clindamycin was detected in 25% of the strains tested. Thirty-four of the macrolide-lincosamide-streptogramin B (MLS(B))-resistant strains carried the erythromycin resistance methylase gene. The results indicate an increase in the prevalence of resistance to MLS(B) and fluoroquinolone antibiotics in C. difficile. Fluoroquinolone resistance is associated with resistance to MLS(B) antimicrobials.     

Antimicrobial-resistant strains of Clostridium difficile from North America

A total of 316 toxigenic Clostridium difficile clinical isolates of known PCR ribotypes from patients in North America were screened for resistance to clindamycin, metronidazole, moxifloxacin, and rifampin. Clindamycin resistance was observed among 16 different ribotypes, with ribotypes 017, 053, and 078 showing the highest proportions of resistance. All isolates were susceptible to metronidazole. Moxifloxacin resistance was present in >90% of PCR-ribotype 027 and 053 isolates but was less common among other ribotypes. Only 7.9% of the C. difficile isolates were resistant to rifampin. Multidrug resistance (clindamycin, moxifloxacin, and rifampin) was present in 27.5% of PCR-ribotype 027 strains but was rare in other ribotypes. These results suggest that antimicrobial resistance in North American isolates of C. difficile varies by strain type and parallels rates of resistance reported from Europe and the Far East.     

Resistance to Moxifloxacin in Toxigenic Clostridium difficile Isolates Is Associated with Mutations in gyrA

Clostridium difficile is the etiological agent of antibiotic-associated colitis and the most common cause of hospital-acquired infectious diarrhea. Fluoroquinolones such as ciprofloxacin are associated with lower risks of C. difficile-associated diarrhea. In this study, we have analyzed 72 C. difficile isolates obtained from patients with different clinical courses of disease, such as toxic megacolon and relapses; the hospital environment; public places; and horses. They were investigated for their susceptibilities to moxifloxacin (MXF), metronidazole (MEO), and vancomycin (VAN). Mutants highly resistant to fluoroquinolones were selected in vitro by stepwise exposure to increasing concentrations of MXF. The resulting mutants were analyzed for the presence of mutations in the quinolone resistance-determining regions of DNA gyrase (gyrA), the production of toxins A and B, and the epidemiological relationship of these isolates. These factors were also investigated using PCR-based methods. All strains tested were susceptible to MEO and VAN. Twenty-six percent of the clinical isolates (19 of 72) were highly resistant to MXF (MIC > or = 16 microg/ml). Fourteen of these 19 strains contained nucleotide changes resulting in amino acid substitutions at position 83 in the gyrA protein. Resistant strains selected in vitro did not contain mutations at that position. These findings indicate that resistance to MXF in a majority of cases may be due to amino acid substitution in the gyrA gene.     

Comparative susceptibilities to fidaxomicin (OPT-80) of isolates collected at baseline, recurrence, and failure from patients in two phase III trials of fidaxomicin against Clostridium difficile infection

A 10-day course of oral fidaxomicin (200 mg twice a day [b.i.d.]), a potent new macrocyclic drug, was compared to vancomycin (125 mg four times a day [q.i.d.]) in 1,164 adults (1,105 in the modified intent-to-treat [mITT] population) with Clostridium difficile infection in two phase III randomized, double-blind trials at sites in North America and 7 European countries. Of 1,105 mITT patients, 792 (71.7%), including 719/999 (72.0%) in the per-protocol (PP) population, provided a C. difficile strain at baseline, of whom 356 received fidaxomicin with 330 cures (92.7%) and 363 received vancomycin with 329 cures (90.6%). The susceptibilities (MIC(90)) of baseline isolates did not predict clinical cure, failure, or recurrence for fidaxomicin (MIC(90), 0.25 μg/ml for both; range, ≤ 0.007 to 1 μg/ml), but there was a one-dilution difference in the MIC(90) (but not the MIC(50)) for vancomycin (MIC(90), 2 μg/ml [range, 0.25 to 8 μg/ml] for cure and 4.0 μg/ml [range, 0.5 to 4 μg/ml] for failures). A total of 65 (7.9%) "rifaximin-resistant" (MIC > 256 μg/ml) strains were isolated in both treatment groups on enrollment, which increased to 25% for failures at the end of therapy. No resistance to either fidaxomicin or vancomycin developed during treatment in either of the phase III studies, although a single strain isolated from a cured patient had an elevated fidaxomicin MIC of 16 μg/ml at the time of recurrence. All isolates were susceptible to ≤ 4 μg/ml of metronidazole. When analyzed by restriction endonuclease analysis (REA) type, 247/719 (34.4%) isolates were BI group isolates, and the MICs were generally higher for all four drugs tested (MIC(90)s: fidaxomicin, 0.5; vancomycin, 2.0; metronidazole, 2.0; and rifaximin, >256 μg/ml) than for the other REA types. There was no correlation between the MIC of a baseline clinical isolate and clinical outcome. MIC(90)s were generally low for fidaxomicin and vancomycin, but BI isolates had higher MICs than other REA group isolates.     

Antimicrobial susceptibility pattern of Clostridium difficile and its relation to PCR ribotypes in a Swedish university hospital

All 238 Clostridium difficile isolates were susceptible to metronidazole and vancomycin, whereas 84% and 1% were resistant to clindamycin and fusidic acid. Etest MICs for metronidazole were lower than agar dilution MICs (P < 0.01) but without difference in susceptible-intermediate-resistant categorization. No particular PCR ribotype was associated with clindamycin or fusidic acid resistance.     

杭州市富阳区住院腹泻患者艰难梭菌感染及菌株分子特征分析

目的了解浙江省杭州市富阳地区住院腹泻患者艰难梭菌感染情况,研究感染患者临床危险因素及分离菌株的分子特征。方法收集2017年5月至2018年2月就诊于第一人民医院的228例腹泻患者的粪便标本,通过厌氧培养进行艰难梭菌菌株分离和鉴定,进行多位点序列基因分型,以琼脂稀释法测定菌株耐药性,对患者临床资料进行统计分析。结果228份标本共检测出产毒型艰难梭菌41株,其中31株为医院获得性腹泻;阳性患者平均年龄67.8岁,主要来自消化内科（34.1%）和肾内科（19.5%）。 ST2、ST3和ST54为主要基因型。 所有菌株对万古霉素、甲硝唑、四环素、哌拉西林敏感。 临床资料统计分析显示,＞60周岁、患基础性疾病和有抗生素用药史的阳性患者数量远高于阴性患者比例（χ2＝4.229、9.022、5.767,P＝0.023、0.001、0.009）。结论杭州市富阳区住院腹泻患者艰难梭菌感染患者主要来源于院内感染。 患有基础性疾病、8周内有抗生素用药史的老年患者为感染的危险因素;在院内感染防控中需进一步加强监测。     

Clostridium difficile colitis that fails conventional metronidazole therapy: response to nitazoxanide

OBJECTIVES: Clostridium difficile-associated disease has increased in incidence and severity. Recommended treatments include metronidazole and vancomycin. Recent investigations, however, document the failure of metronidazole to cure a substantial proportion of patients with Clostridium difficile colitis, but oral administration of vancomycin raises concerns over selection of antibiotic-resistant organisms in the hospital environment. We have recently shown that nitazoxanide is as effective as metronidazole in initial therapy for C. difficile colitis. We hypothesized that this drug might be effective in treating patients who fail therapy with metronidazole. METHODS: In the present study, we identified 35 patients who failed treatment with metronidazole for C. difficile colitis; failure was defined as either no improvement in symptoms or signs of disease (28 patients) after >or= 14 days of treatment with metronidazole or prompt recurrence on at least two occasions after initially responding to such treatment (seven patients). These patients were ill with numerous co-morbidities. Nitazoxanide, 500 mg twice daily, was given for 10 days; results from all patients are included. RESULTS: Twenty-six (74%) of 35 patients responded to nitazoxanide, of whom seven later had recurrent disease, yielding a cure rate of 19 of 35 (54%) from initial therapy. Three who initially failed and one who had recurrent disease were re-treated with, and responded to, nitazoxanide. Thus, the aggregate cure with nitazoxanide in this difficult-to-treat population was 23 of 35 (66%). CONCLUSIONS: Nitazoxanide appears to provide effective therapy for patients with C. difficile colitis who fail treatment with metronidazole.     

Antimicrobial susceptibilities of Gardnerella vaginalis.

The in vitro susceptibilities of 93 clinical isolates of Gardnerella vaginalis to 25 antimicrobial agents were determined by the agar dilution method. All isolates were susceptible to penicillin, ampicillin, erythromycin, clindamycin, chloramphenicol, and trimethoprim. Activity was poor for vancomycin, LY146032, the cephalosporins, ciprofloxacin, and imipenem. Some resistance was observed with tetracycline and minocycline. The MICs of metronidazole paralleled those of tinidazole, with the hydroxymetabolite of metronidazole being the most active. One strain was resistant to all three agents. Marked resistance to aztreonam, amikacin, and sulfamethoxazole was observed.     

5岁以下腹泻患者分离的艰难梭菌分子型别和耐药特征分析

目的 分析中国部分地区5岁以下腹泻患儿分离的艰难梭菌的分子型别和耐药特征,为后续研究及临床用药提供数据支持.方法 收集2010-2020年云南、山东、上海、陕西、河南和北京6省市5岁以下腹泻患儿的粪便标本,通过厌氧培养分离和鉴定艰难梭菌.使用聚合酶链式反应(PCR)方法进行毒素基因检测、多位点序列分型(MLST)和核糖...     

Susceptibility testing of Clostridium difficile against metronidazole and vancomycin by disk diffusion and Etest.

A prospective study on the susceptibility of Clostridium difficile to metronidazole and vancomycin using the Etest and disk diffusion test was performed over a 6-month period. One hundred strains were tested; one strain was highly resistant to metronidazole (MIC = 64 micrograms/mL). The zone size of inhibition by the disk diffusion test correlated with the MIC as determined by the Etest (regression coefficient = -0.043 for metronidazole and -0.044 for vancomycin, p < 0.001 for both antibiotics). However, the correlation coefficient was low for both metronidazole (r = 0.574) and vancomycin (r = 0.473); hence the zone of inhibition by disk diffusion test could not predict the MIC satisfactorily. Metronidazole is still the first-line antibiotic for the treatment of C. difficile-associated diarrhea because the incidence of metronidazole resistant strains remains very low. However, the efficacy of metronidazole in the treatment of infections attributable to isolates with high-level metronidazole resistance may be compromised because the fecal concentration of metronidazole is relatively low when compared with the MIC values of the less susceptible strains. Oral vancomycin is the drug of choice under such circumstances, as its fecal concentration is much higher than that of metronidazole.     

Antimicrobial phenotypes and molecular basis in clinical strains of Clostridium difficile

Clostridium difficile remains the leading cause of nosocomial-acquired diarrhea. This study investigated antimicrobial susceptibility patterns of C. difficile over a 3-year period. Three hundred seventeen C. difficile isolates recovered between 2002 and 2004 were analyzed for their susceptibility to erythromycin (ERY), clindamycin (CLI), moxifloxacin (MXF), doxycycline (DOX), vancomycin (VAN), and metronidazole (MTR) by Etest. The molecular basis for resistance was investigated using polymerase chain reaction (PCR) and DNA sequencing. PCR ribotyping was used to differentiate strains. All strains were susceptible to VAN and MTR. Resistance rates to ERY/CLI, MXF, and DOX increased during the study period. Eighty-four (26.5%) strains exhibited resistance against ERY/CLI, MXF, and DOX. Prevalence of resistance genes was as follows: ermB, 83; ermQ, 0; ermFS, 1; tetM, 84; tetP, 0; tetO, 2; and gyrA mutation, 76. These results indicate an increasing trend in the prevalence of combined resistance against macrolide-lincosamide-streptogramin B antibiotics, fluoroquinolones, and tetracycline in C. difficile. The lack of understanding of antibiotic resistance mechanisms in C. difficile and the increased resistant strains warrants further investigations.     

Antimicrobial Susceptibilities and Serogroups of Clinical Strains of Clostridium difficile Isolated in France in 1991 and 1997

Glycopeptides (vancomycin and teicoplanin) and metronidazole are the drugs of choice for the treatment of Clostridium difficile infections, but trends in susceptibility patterns have not been assessed in the past few years. The objective was to study the MICs of glycopeptides and metronidazole for unrelated C. difficile strains isolated in 1991 (n = 100) and in 1997 (n = 98) by the agar macrodilution, the E-test, and the disk diffusion methods. Strain susceptibilities to erythromycin, clindamycin, tetracycline, rifampin, and chloramphenicol were also determined by the ATB ANA gallery (bioMérieux, La Balme-les-Grottes, France). The MICs at which 50% of isolates are inhibited (MIC(50)s) and MIC(90)s of glycopeptides and metronidazole remained stable between 1991 and 1997. All the strains were inhibited by concentrations that did not exceed 2 microgram/ml for vancomycin and 1 microg/ml for teicoplanin. Comparison of MICs determined by the agar dilution method recommended by the National Committee for Clinical Laboratory Standards and the E test showed correlations (+/-2 dilutions) of 86. 6, 95.9, and 99% for metronidazole, vancomycin, and teicoplanin, respectively. The E test always underestimated the MICs. Strains with decreased susceptibility to metronidazole (MICs, >/=8 microgram/ml) were isolated from six patients (n = 4 in 1991 and n = 2 in 1997). These strains were also detected by the disk diffusion method (zone inhibition diameter, /=1 microgram/ml), clindamycin (MICs, >/=2 microgram/ml), tetracycline (MICs, >/=8 microgram/ml), rifampin (MICs, >/=4 microgram/ml), and chloramphenicol (MICs, >/=16 microgram/ml) was observed in 64.2, 80.3, 23.7, 22.7, and 14.6% of strains, respectively. Strains isolated in 1997 were more susceptible than those isolated in 1991, and this trend was correlated to a major change in serogroup distribution. Periodic studies are needed in order to detect changes in serogroups and the emergence of strains with decreased susceptibility to therapeutic drugs.     

In vitro and in vivo activities of nitazoxanide against Clostridium difficile

We have used the hamster model of antibiotic-induced Clostridium difficile intestinal disease to evaluate nitazoxanide (NTZ), a nitrothiazole benzamide antimicrobial agent. The following in vitro and in vivo activities of NTZ in the adult hamster were examined and compared to those of metronidazole and vancomycin: (i) MICs and minimum bactericidal concentrations (MBCs) against C. difficile, (ii) toxicity, (iii) ability to prevent C. difficile-associated ileocecitis, and (iv) propensity to induce C. difficile-associated ileocecitis. The MICs and MBCs of NTZ against 15 toxigenic strains of C. difficile were comparable to those of vancomycin or metronidazole. C. difficile-associated ileocecitis was induced with oral clindamycin and toxigenic C. difficile in a group of 60 hamsters. Subgroups of 10 hamsters were given six daily intragastric treatments of NTZ (15, 7.5, and 3.0 mg/100 g of body weight [gbw]), metronidazole (15 mg/100 gbw), vancomycin (5 mg/100 gbw), or saline (1 ml/100 gbw). Animals receiving saline died 3 days post-C. difficile challenge. During the treatment period, NTZ (>/=7.5 mg/100 gbw), like metronidazole and vancomycin, prevented outward manifestations of clindamycin-induced C. difficile intestinal disease. Six of ten hamsters on a scheduled dose of 3.0 mg of NTZ/100 gbw survived for the complete treatment period. Of these surviving animals, all but three died of C. difficile disease by between 3 and 12 days following discontinuation of antibiotic therapy. Another group of hamsters received six similar daily doses of the three antibiotics, followed by an inoculation with toxigenic C. difficile. All of the NTZ-treated animals survived the 15-day postinfection period. Upon necropsy, all hamsters appeared normal: there were no gross signs of toxicity or C. difficile intestinal disease, nor was C. difficile detected in the cultures of the ceca of these animals. By contrast, vancomycin and metronidazole treatment induced fatal C. difficile intestinal disease in 20 and 70% of recipients, respectively.     

Antimicrobial susceptibility of Clostridium difficile determined by disc diffusion and breakpoint methods

The susceptibility of 160 isolates of Clostridium difficile to eight antimicrobial agents was studied by two methods. There was generally good agreement between the results obtained with the disc diffusion and breakpoint methods. More than 90% of isolates studied were considered sensitive by both methods. However there was a major difference between the results obtained with the two methods for penicillin G and clindamycin, resistance to both agents being overestimated by the disc diffusion method. Several isolates exhibited multiple resistance to penicillin G, tetracycline, erythromycin and clindamycin. All isolates were inhibited by both metronidazole and vancomycin. None of the isolates produced beta-lactamase.     

Characterizations of clinical isolates of clostridium difficile by toxin genotypes and by susceptibility to 12 antimicrobial agents, including fidaxomicin (OPT-80) and rifaximin: a multicenter study in Taiwan

A total of 403 nonduplicate isolates of Clostridium difficile were collected at three major teaching hospitals representing northern, central, and southern Taiwan from January 2005 to December 2010. Of these 403 isolates, 170 (42.2%) were presumed to be nontoxigenic due to the absence of genes for toxins A or B or binary toxin. The remaining 233 (57.8%) isolates carried toxin A and B genes, and 39 (16.7%) of these also had binary toxin genes. The MIC(90) of all isolates for fidaxomicin and rifaximin was 0.5 μg/ml (range, ≤ 0.015 to 0.5 μg/ml) and >128 μg/ml (range, ≤ 0.015 to >128 μg/ml), respectively. All isolates were susceptible to metronidazole (MIC(90) of 0.5 μg/ml; range, ≤ 0.03 to 4 μg/ml). Two isolates had reduced susceptibility to vancomycin (MICs, 4 μg/ml). Only 13.6% of isolates were susceptible to clindamycin (MIC of ≤ 2 μg/ml). Nonsusceptibility to moxifloxacin (n = 81, 20.1%) was accompanied by single or multiple mutations in gyrA and gyrB genes in all but eight moxifloxacin-nonsusceptible isolates. Two previously unreported gyrB mutations might independently confer resistance (MIC, 16 μg/ml), Ser416 to Ala and Glu466 to Lys. Moxifloxacin-resistant isolates were cross-resistant to ciprofloxacin and levofloxacin, but some moxifloxacin-nonsusceptible isolates remained susceptible to gemifloxacin or nemonoxacin at 0.5 μg/ml. This study found the diversity of toxigenic and nontoxigenic strains of C. difficile in the health care setting in Taiwan. All isolates tested were susceptible to metronidazole and vancomycin. Fidaxomicin exhibited potent in vitro activity against all isolates tested, while the more than 10% of Taiwanese isolates with rifaximin MICs of ≥ 128 μg/ml raises concerns.