Resistance Of Bacteroides Research Articles

Rapidly Growing Aneurysm with Ulcer-like Projection Complicated with Bacteroides ovatus Bacteremia

The incidence of anaerobic bacteremia has been increasing over the past several decades. Further, antibiotic resistance in Bacteroides is a pertinent issue. A 76-year-old man was brought to our hospital with complaints of fever, chills, and abdominal pain. Empiric antibiotics induced minimal relief. The blood culture was positive for multi-drug resistant Bacteroides ovatus. Our patient developed a periaortic abscess in the abdominal aorta and a thoracic aortic aneurysm with ulcer-like projection (ULP), which rapidly increased in size. He was transferred to the tertiary medical institution for surgical drainage. This case suggests that bacteremia can exacerbate aneurysms with ulcerative lesions. Anaerobic bacteremia is a possible differential diagnosis when periaortic abscess formation is present. Early surgical consultation and appropriate antibiotic selection are crucial in anaerobic bacteremia treatment.

Five years prospective survey of antibiotic resistance in Bacteroides and Parabacteroides isolated from inpatients of clinical hospital in Warsaw, Poland

Objectives: Diagnostics of anaerobic bacterial infections and determination of drug susceptibility are technically difficult and time-consuming; therefore, the number of studies on Anaerobic Gram-negative bacilli is significantly limited, especially in Europe. The aim of the study was to analyze the antibiotic susceptibility of clinically important anaerobic bacteria Bacteroides spp. and Parabacteroides distasonis. Strains were isolated from infections of patients hospitalized at one Polish hospital as a result of routine microbiological diagnostics. Material and methods: Clinical isolates were identified with MALDI-TOF MS. Antimicrobial susceptibility of 276 strains was carried out by E-test gradient strip to commonly used antibiotics i.e. benzylpenicillin, amoxicillin with clavulanic acid, imipenem, clindamycin and metronidazole. MIC values were determined. The interpretation of antimicrobial susceptibility tests were conducted in accordance with The European Committee on Antimicrobial Susceptibility Testing (EUCAST) recommendations. Results: Susceptibility tests of all isolates yielded the following rates of resistance to the evaluated β-lactam antibiotics: benzylpenicillin (96%), amoxicillin/clavulanic acid (7.6%), imipenem (2.1%). In presented study 38.8% of clindamycin-resistant strains were isolated, among them 18.3% of B. fragilis and 53.85% P. distasonis. All strain were susceptible to metronidazole. Conclusions: Obtained results and analysis of the results of other researchers convinces us that it is necessary to routinely or at least periodically monitor drug susceptibility of clinical isolates of anaerobic bacteria and use targeted therapy based on the result of the antibiogram. Although high percentage of the tested Bacteroides and Parabacteroides strains remained susceptible to metronidazole and β-lactam antibiotics the use of clindamycin in empirical therapy may not be efficacious. Antibiogram results should be consult with the staff responsible for patient treatment and hospital antibiotic policies.

Emerging metronidazole resistance in Bacteroides spp. and its association with the nim gene: a study from North India.

Bacteroides spp. are anaerobic resident intestinal flora but are also known to cause severe morbidity. They are resistant to a wide variety of antimicrobial agents, including metronidazole, which has been shown to be associated with specific nitroimidazole (nim) resistance genes. Metronidazole resistance is emerging worldwide, although presently it remains at ca. 5%. This study aimed to determine the metronidazole susceptibility and distribution of nim genes in Bacteroides spp. clinical isolates in India. The relationship among strains harbouring nim genes and their susceptibility to metronidazole was also analysed. A total of 42 Bacteroides spp. clinical isolates were identified using an advanced MALDI-TOF system. Minimum inhibitory concentrations (MICs) for metronidazole were determined by the agar dilution method. Bacterial DNA was extracted and was subjected to nim gene PCR and the amplified PCR products were sequenced to determine the prevalent nim types. Bacteroides fragilis was the most common isolate (64%) among all Bacteroides spp. isolates. Among the total 42 clinical Bacteroides spp. isolates, 29 (69%) were susceptible and 13 (31%) were resistant to metronidazole by the agar dilution method. nim gene PCR performed on 38 isolates showed positivity in 20 isolates (53%), of which 12 had high metronidazole MICs (χ2 test, P<0.005). On sequencing, these nim genes were most closely related to nimE type. Resistance to metronidazole is consistently emerging worldwide. There is a significant association of the nim gene with metronidazole resistance. Periodic surveillance is needed to detect geographic and temporal trends in nim gene prevalence.

Phenotypic and Genotypic Characterization of Multidrug-ResistantBacteroides,Parabacteroidesspp., andPseudoflavonifractorfrom a Costa Rican Hospital

Multidrug resistance in Bacteroides spp. and related genera is uncommon and has not been described in Latin America until now. We studied phenotypically and genotypically the multidrug resistance of 10 clinical strains of Bacteroides, two of Parabacteroides distasonis, and one of Pseudoflavonifractor capillosus recovered in a national hospital between 2006 and 2010. To this end, we determined minimum inhibitory concentrations (MICs) of amoxicillin, amoxicillin-clavulanic acid, cefotaxime, imipenem, clindamycin, ciprofloxacin, tetracycline, and metronidazole using E-tests, evaluated the isolates for β-lactamases with nitrocefin hydrolysis tests, performed a polymerase chain reaction (PCR)-based screening of erm, tet, and nim genes, obtained partial gyrA sequences, and studied the effect of tazobactam and efflux pump inhibitors (EPI) on the MIC of cefotaxime, clindamycin, and ciprofloxacin. Three isolates were resistant to four different classes of antibiotics and 10 were resistant to three. β-lactam resistance was in most cases due to β-lactamases susceptible of partial inhibition by tazobactam. Ten isolates were cfxA-positive and two isolates had cepA. Twelve isolates were highly resistant to clindamycin and nine were highly resistant to ciprofloxacin. However, these phenotypes were not linked to ermA, ermB, ermF, and ermG or mutations in gyrA. Addition of EPI lowered the MICs of clindamycin and ciprofloxacin of one and four isolates, respectively. Twelve isolates had tetQ and four were positive for tetM. In both cases, genes of the two-component system RteAB accompanied tet genes. Although metronidazole susceptibility was universal, nim genes were not present. To our knowledge, this is the first report of multidrug resistance due to less commonly identified or alternative mechanisms in strains of Bacteroides and related species from a developing country.

Prevalence of in vitro susceptibility to carbapenems among clinical isolates of the Bacteroides fragilis group from a Colombian tertiary hospital.

Clinical isolates of the Bacteroides fragilis group (n=29) were collected from patients attending a tertiary care hospital in Medellin, Colombia, during 2012 and were tested for susceptibility to four carbapenems using the Clinical and Laboratory Standards Institute (CLSI) Etest method. Overall rates of percent susceptibility were as follows: meropenem, 93.1%; imipenem, 100%; ertapenem, 96.6%; and doripenem, 93.1%. This is the first study to show rates of resistance of Bacteroides spp. to carbapenems in Colombia among a species formerly thought to be fully susceptible to this type of antimicrobial.

Extended role for insertion sequence elements in the antibiotic resistance ofBacteroides

Extended role for insertion sequence elements in the antibiotic resistance of<i>Bacteroides</i>

Differentiation of cfiA -Negative and cfiA -Positive Bacteroides fragilis Isolates by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

Carbapenem resistance in Bacteroides fragilis is associated with cfiA-encoded class B metallo-beta-lactamase. cfiA-negative and cfiA-positive isolates belong to genotypically distinct groups. Of a total of 248 B. fragilis isolates included in this study, 214 were susceptible, 10 were intermediate, and 24 were resistant to meropenem. We show that matrix-assisted laser desorption ionization-time of flight mass spectrometry is able to differentiate between cfiA-negative and cfiA-positive isolates and predict carbapenem resistance in a routine laboratory setting.

Antimicrobial Susceptibility Patterns for Recent Clinical Isolates of Anaerobic Bacteria in South Korea

We determined the antimicrobial susceptibilities of 255 clinical isolates of anaerobic bacteria collected in 2007 and 2008 at a tertiary-care hospital in South Korea. Piperacillin-tazobactam, cefoxitin, imipenem, and meropenem were highly active beta-lactam agents against most of the isolates tested. The rates of resistance of Bacteroides fragilis group organisms and anaerobic gram-positive cocci to moxifloxacin were 11 to 18% and 0 to 27%, respectively.

Lessons Learned from the Anaerobe Survey: Historical Perspective and Review of the Most Recent Data (2005–2007)

The rationale and lessons learned through the evolution of the National Survey for the Susceptibility of Bacteroides fragilis Group from its initiation in 1981 through 2007 are reviewed here. The survey was conceived in 1980 to track emerging antimicrobial resistance in Bacteroides species. Data from the last 11 years of the survey (1997-2007), including 6574 isolates from 13 medical centers, were analyzed for in vitro antimicrobial resistance to both frequently used and newly developed anti-anaerobic agents. The minimum inhibitory concentrations of the antibiotics were determined using agar dilution in accordance with Clinical and Laboratory Standards Institute recommendations. The analyses revealed that the carbapenems (imipenem, meropenem, ertapenem, and doripenem) and piperacillin-tazobactam were the most active agents against these pathogens, with resistance rates of 0.9%-2.3%. In the most recent 3 years of the survey (2005-2007), resistance to some agents was shown to depend on the species, such as ampicillin-sulbactam against Bacteroides distasonis (20.6%) and tigecycline against Bacteroides uniformis and Bacteroides eggerthii ( approximately 7%). Very high resistance rates (>50%) were noted for moxifloxacin and trovafloxacin, particularly against Bacteroides vulgatus. During that period of study, non-B. fragilis Bacteroides species had >40% resistance to clindamycin. Metronidazole-resistant Bacteroides strains were also first reported during that period. In summary, resistance to antibiotics was greater among non-B. fragilis Bacteroides species than among B. fragilis and was especially greater among species with a low frequency of isolation, such as Bacteroides caccae and B. uniformis. The emergence of resistance among the non-B. fragilis Bacteroides species underscores the need for speciation of B. fragilis group isolates and for clinicians to be aware of associations between species and drug resistance.

Metronidazole resistance in Bacteroides spp. carrying nim genes and the selection of slow-growing metronidazole-resistant mutants.

Human clinical isolates of Bacteroides spp. originating from patients in the UK were investigated for the presence of metronidazole resistance determinants (nim genes) and their presence was related to the MIC of metronidazole for the isolates. Isolates were screened for susceptibility to a metronidazole disc and had their MIC determined by the Etest method. They were investigated for the presence of nim genes by PCR. An experiment to determine the effect of prolonged exposure to metronidazole was applied to nim-positive isolates with MICs below the therapeutic breakpoint. Fifty of 206 isolates (24%) were found to possess nim genes and these had MICs of metronidazole ranging from 1.5 to >256 mg/L with 24 (11.6%) above the therapeutic breakpoint of 16 mg/L. The remaining 26 nim-gene-positive isolates had MICs that were still below the therapeutic breakpoint, ranging from 1.5 to 6.0 mg/L. nim genes were not found in 156 (76%) isolates, and all but seven of these were susceptible to a 5 microg disc of metronidazole. Ten members of the group for which the MICs were below the therapeutic level were found to have slow-growing sub-populations with metronidazole MICs ranging from 8.0 to >256 mg/L that became evident after prolonged exposure to metronidazole in vitro. This resistance selection process was sometimes reversible after passage in the absence of metronidazole; however, seven of the 10 slow-growing mutants converted to stable high-level resistance (MIC >256 mg/L). Although the presence of nim genes per se does not always equate to therapeutic resistance, and other metronidazole resistance mechanisms may exist, this study has shown that prolonged exposure of nim-gene-carrying Bacteroides spp. to metronidazole can select for therapeutic resistance.

Plasmid-mediated complementation of gyrA and gyrB in fluoroquinolone-resistant Bacteroides fragilis.

To identify whether mutations in gyrA and gyrB confer fluoroquinolone resistance in Bacteroides fragilis. Eight fluoroquinolone-resistant (FQR) strains were complemented with plasmid-mediated B. fragilis wild-type gyrA (pMP1) and gyrB (pMP2), and MICs determined. Sequence analysis of the gyrA and gyrB quinolone resistance determining region (QRDR) was performed for all strains. MICs of fluoroquinolones were two- to 32-fold higher than wild-type for all mutants. Five mutants had a substitution in GyrA (Ser-82-->Phe), one mutant had a substitution in GyrA (Asp-81-->Gly), one mutant had a substitution in GyrB (Glu-478-->Lys), and one resistant strain did not contain mutations in the QRDR of gyrA or gyrB. Following complementation with pMP1 or pMP2, the MICs of fluoroquinolones were reduced two- to 32-fold for the mutants. These studies verify that substitutions in GyrA and GyrB confer resistance in B. fragilis. Other mechanisms are also responsible for resistance since not all resistant strains fully complemented to the wild-type phenotype.

Resistance to beta-lactam antibiotics in Bacteroides spp.

Bacteroides spp., particularly B. fragilis, are well-recognised bacterial pathogens. Production of the typical beta-lactamases of Bacteroides restricts the therapeutic use of beta-lactam agents mainly to the beta-lactamase inhibitor combinations and carbapenems. These compounds have the advantage of broad-spectrum activity and the ability to combat polymicrobial infections. Resistance of Bacteroides spp. to beta-lactam antibiotics appears to be increasing, largely because of an overall increase in beta-lactamase activity. There has been a rise in the prevalence of isolates showing high-level production of typical Bacteroides beta-lactamases and an increase in reports other potent beta-lactamase types. In the case of B. fragilis, metallo-enzymes are a particular threat to current therapeutic practice, as they are not inhibited by common beta-lactamase inhibitors and are able to hydrolyse carbapenems. The presence of permeability barriers may confer low-level beta-lactam resistance and supplement the effect of beta-lactamase activity. There are also sporadic reports of loss of beta-lactam activity because of reduced affinity of the penicillin-binding proteins.

Lactate dehydrogenase activity as a cause of metronidazole resistance in Bacteroides fragilis NCTC 11295

Enzymes acting on pyruvate as a parameter of the ATP regeneration system were studied as a cause of metronidazole resistance in Bacteroides fragilis NCTC 11295. The resistant strain had higher lactate dehydrogenase activity and produced more lactate than susceptible strains, suggesting that the enzyme is more active in lactic acid fermentation. Furthermore, the reaction catalysed by lactate dehydrogenase occurred up to 48 mg/L metronidazole, whereas the reaction catalysed by pyruvate: ferredoxin oxidoreductase reaction stopped at 2 mg/L. The mechanism of metronidazole resistance in B. fragilis NCTC 11295 may be due to the high activity of lactate dehydrogenase which compensates for the decreased activity of pyruvate: ferredoxin oxidoreductase in the presence of metronidazole.

Mechanisms of tetracycline and rifampicin resistance inBacteroides fragilis mutants obtained by ionizing radiation

Based on a dose-survival curve, a radiation dose of 3.99 C/kg was used to induce antibiotic-resistant mutants inBacteroides fragilis. Escherichia coli B/r membrane fragments were employed as a reducing agent. Antibiotic-resistant mutants ofB. fragilis were utilized to study the mechanism by which these organisms become resistant to selected chemotherapeutic agents. Decreased accumulation of tetracycline by resistant mutants ofB. fragilis suggests that the resistance to this antibiotic is associated with the outer membrane permeability. There is a marked difference in the inhibitory action of rifampicin on RNA polymerase activity in rifampicin-sensitive and-resistant strains ofB. fragilis. This enzyme is, therefore, the likely target for inhibition of bacterial growth in this anaerobe by rifampicin.

Antibiotic resistance in anaerobic bacteria.

An improved understanding of the mechanisms of resistance and transfer in Bacteroides fragilis has been gained over the past decade. B. fragilis resistance to most penicillins is largely due to production of chromosomal beta-lactamases, although recent findings indicate these micro-organisms can acquire novel beta-lactamases, which can even inactivate imipenem. Several of the beta-lactamases of B. fragilis are transferable. Most of our understanding of transferable antimicrobial resistance in Bacteroides spp. has been gained through studies of the clindamycin-erythromycin resistance determinant found on pBFTM10. This resistance is encoded on a transposon and is widely distributed among the naturally occurring clindamycin-resistant isolates. Recent studies have shown that DNA can be transferred from Escherichia coli to B. fragilis, from one B. fragilis to another, and from B. fragilis back to E. coli. Data from a large survey in the United States indicate that susceptibility patterns of B. fragilis differ in each of the eight participating centres. Overall, piperacillin and cefoxitin have been found to be the most active beta-lactam agents.

Antimicrobial resistance in Bacteroides.

Antimicrobial resistance in Bacteroides.

Expression in Escherichia coli of cryptic tetracycline resistance genes from Bacteroides R plasmids

The putative clindamycin resistance region of the Bacteroides fragilis R plasmid pBF4 was cloned in the vector R300B in Escherichia coli. This 3.8-kb EcoRI D fragment from pBF4 expressed noninducible tetracycline resistance in E. coli under aerobic but not anaerobic growth conditions. The fragment does not express tetracycline resistance in Bacteroides, a strict anaerobe. The separate tetracycline resistance transfer system in the Bacteroides host strain V479-1 has no homology to the cryptic determinant on pBF4. In addition, this aerobic tetracycline resistance determinant is not homologous to the three major plasmid mediated tetracycline resistance regions found in facultative gram-negative bacteria, represented by R100, RK2, and pBR322. A similar cryptic tetracycline resistance fragment was cloned from pCP1, a separate clindamycin resistance plasmid from Bacteroides that shares homology with the EcoRI D fragment of pBF4. This study identifies cryptic drug resistance determinants in Bacteroides that are expressed when inserted into an aerobically growing organism.

Activity of cefoperazone and two beta-lactamase inhibitors, sulbactam and clavulanic acid, against Bacteroides spp. correlated with beta-lactamase production.

A total of 102 isolates of Bacteroides spp. were studied for beta-lactamase production and susceptibility to cefoperazone alone or in combination with either of the beta-lactamase inhibitors sulbactam and clavulanic acid. The geometric mean minimal inhibitory concentration of cefoperazone alone was 31.5 micrograms/ml and when combined with 10 micrograms of sulbactam per ml or 2 micrograms of clavulanic acid per ml was reduced to 5.4 and 9.2 micrograms/ml, respectively. When bacterial suspensions were tested for beta-lactamase production with nitrocefin, 91 (89.2%) of these isolates produced the enzyme. The geometric mean minimal inhibitory concentrations of cefoperazone rose only slightly for isolates with low or intermediate enzyme activity but rose significantly for those with high activity. The addition of EDTA to cefoperazone significantly more frequently enhanced the activity of cefoperazone against beta-lactamase-negative as opposed to beta-lactamase-positive isolates. Furthermore, EDTA resulted in synergistic activity of the cefoperazone-sulbactam combination on beta-lactamase-positive isolates for which the combination had previously not shown a synergistic effect. This study demonstrates the relationship between beta-lactamase production and the resistance of Bacteroides spp. to cefoperazone and shows that inhibition of these enzymes can reverse this resistance.

Transferable plasmid-mediated antibiotic resistance in Bacteroides.

Plasmid-mediated resistance to chloramphenicol (Chlr), erythromycin (Eryr), tetracycline (Tetr) and clindamycin (Clindr) was transferred from three clinical isolates of Bacteroides fragilis and one faecal isolate of B. thetaiotaomicron to strains of B. fragilis, B. distasonis and Escherichia coli, and subsequently to B. fragilis and E. coli second-and third-stage recipients in series. Successful transfer was achieved by membrane-filter and centrifugation techniques that provide stable cell-to-cell contact but not by simple mixed broth culture. Chlr Eryr Tetr and Clindr Eryr were transferred at high frequency (1.9 x 10(-3)-1.8 x 10(-4)) but Tetr was transferred at low frequencies (1-1.6 x 10(-6)). Segregation of resistance markers was observed with selection for Tetr when donors were Chlr Eryr Tetr and Chlr Tetr. All transcipients were identical with the parent recipient strains but had the resistance markers of the donor strains. Resistance to antibiotics other than tetracycline was cured by growth with subinhibitory concentrations of aminoacridines and ethidium bromide for 24 h; cure of solitary Tetr required longer incubation (21 days). Identical plasmid DNA bands were demonstrated by agarose-gel electrophoresis in all the donor and corresponding transcipient strains but plasmids were not found in the recipient strains or in strains cured of resistance. Plasmid-mediated transferable antiobiotic resistance in Bacteroides spp. may compromise the treatment of infections and may provide a reservoir of antibiotic resistance in the intestinal flora.

Non-plasmid associated transfer of antibiotic resistance in Bacteroides.

Non-plasmid associated transfer of antibiotic resistance in Bacteroides.