scholarly journals Epidemiology of Conjugative Plasmid-Mediated AmpC β-Lactamases in the United States

2004 ◽  
Vol 48 (2) ◽  
pp. 533-537 ◽  
Author(s):  
M. Alvarez ◽  
J. H. Tran ◽  
N. Chow ◽  
G. A. Jacoby

ABSTRACT A sample of 752 resistant Klebsiella pneumoniae, Klebsiella oxytoca, and Escherichia coli strains from 70 sites in 25 U.S. states and the District of Columbia was examined for transmissibility of resistance to ceftazidime and the nature of the plasmid-mediated β-lactamase involved. Fifty-nine percent of the K. pneumoniae, 24% of the K. oxytoca, and 44% of the E. coli isolates transferred resistance to ceftazidime. Plasmids encoding AmpC-type β-lactamase were found in 8.5% of the K. pneumoniae samples, 6.9% of the K. oxytoca samples, and 4% of the E. coli samples, at 20 of the 70 sites and in 10 of the 25 states. ACT-1 β-lactamase was found at eight sites, four of which were near New York City, where the ACT-1 enzyme was first discovered; ACT-1 β-lactamase was also found in Massachusetts, Pennsylvania, and Virginia. FOX-5 β-lactamase was also found at eight sites, mainly in southeastern states but also in New York. Two E. coli strains produced CMY-2, and one K. pneumoniae strain produced DHA-1 β-lactamase. Pulsed-field gel electrophoresis and plasmid analysis suggested that AmpC-mediated resistance spread both by strain and plasmid dissemination. All AmpC β-lactamase-containing isolates were resistant to cefoxitin, but so were 11% of strains containing transmissible SHV- and TEM-type extended-spectrum β-lactamases. A β-lactamase inhibitor test was helpful in distinguishing the two types of resistance but was not definitive since 24% of clinical isolates producing AmpC β-lactamase had a positive response to clavulanic acid. Coexistence of AmpC and extended-spectrum β-lactamases was the main reason for these discrepancies. Plasmid-mediated AmpC-type enzymes are thus responsible for an appreciable fraction of resistance in clinical isolates of Klebsiella spp. and E. coli, are disseminated around the United States, and are not so easily distinguished from other enzymes that mediate resistance to oxyimino-β-lactams.

2019 ◽  
Vol 6 (7) ◽  
Author(s):  
Ayesha Khan ◽  
Truc T Tran ◽  
Rafael Rios ◽  
Blake Hanson ◽  
William C Shropshire ◽  
...  

Abstract Background Treatment of serious infections due to multidrug-resistant (MDR) Pseudomonas aeruginosa remains a challenge, despite the introduction of novel therapeutics. In this study, we report 2 extensively drug-resistant clinical isolates of sequence type (ST) 309 P aeruginosa resistant to all β-lactams, including the novel combinations ceftolozane/tazobactam, ceftazidime/avibactam, and meropenem/vaborbactam. Methods Isolates were sequenced using both short-read (Illumina) and long-read technology to identify resistance determinants, polymorphisms (compared with P aeruginosa PAO1), and reconstruct a phylogenetic tree. A pair of β-lactamases, Guiana extended spectrum β-lactamase (GES)-19 and GES-26, were cloned and expressed in a laboratory strain of Escherichia coli to examine their relative impact on resistance. Using cell lysates from E coli expressing the GES genes individually and in tandem, we determined relative rates of hydrolysis for nitrocefin and ceftazidime. Results Two ST309 P aeruginosa clinical isolates were found to harbor the extended spectrum β-lactamases GES-19 and GES-26 clustered in tandem on a chromosomal class 1 integron. The presence of both enzymes in E coli was associated with significantly elevated minimum inhibitory concentrations to aztreonam, cefepime, meropenem, ceftazidime/avibactam, and ceftolozane/tazobactam, compared with those expressed individually. The combination of ceftazidime/avibactam plus aztreonam was active in vitro and used to achieve cure in one patient. Phylogenetic analysis revealed ST309 P aeruginosa are closely related to MDR strains from Mexico also carrying tandem GES. Conclusions The presence of tandem GES-19 and GES-26 is associated with resistance to all β-lactams, including ceftolozane/tazobactam. Phylogenetic analysis suggests that ST309 P aeruginosa may be an emerging threat in the United States.


2011 ◽  
Vol 55 (12) ◽  
pp. 5666-5675 ◽  
Author(s):  
Bashar W. Shaheen ◽  
Rajesh Nayak ◽  
Steven L. Foley ◽  
Ohgew Kweon ◽  
Joanna Deck ◽  
...  

ABSTRACTResistance to extended-spectrum cephalosporins (ESC) among members of the familyEnterobacteriaceaeoccurs worldwide; however, little is known about ESC resistance inEscherichia colistrains from companion animals. Clinical isolates ofE. coliwere collected from veterinary diagnostic laboratories throughout the United States from 2008 to 2009.E. coliisolates (n= 54) with reduced susceptibility to ceftazidime or cefotaxime (MIC ≥ 16 μg/ml) and extended-spectrum-β-lactamase (ESBL) phenotypes were analyzed. PCR and sequencing were used to detect mutations in ESBL-encoding genes and the regulatory region of the chromosomal geneampC. Conjugation experiments and plasmid identification were conducted to examine the transferability of resistance to ESCs. All isolates carried theblaCTX-M-1-group β-lactamase genes in addition to one or more of the following β-lactamase genes:blaTEM,blaSHV-3,blaCMY-2,blaCTX-M-14-like, andblaOXA-1.DifferentblaTEMsequence variants were detected in some isolates (n= 40). Three isolates harbored ablaTEM-181gene with a novel mutation resulting in an Ala184Val substitution. Approximately 78% of the isolates had mutations in promoter/attenuator regions of the chromosomal geneampC, one of which was a novel insertion of adenine between bases −28 and −29. Plasmids ranging in size from 11 to 233 kbp were detected in the isolates, with a common plasmid size of 93 kbp identified in 60% of isolates. Plasmid-mediated transfer of β-lactamase genes increased the MICs (≥16-fold) of ESCs for transconjugants. Replicon typing among isolates revealed the predominance of IncI and IncFIA plasmids, followed by IncFIB plasmids. This study shows the emergence of conjugative plasmid-borne ESBLs amongE. colistrains from companion animals in the United States, which may compromise the effective therapeutic use of ESCs in veterinary medicine.


1996 ◽  
Vol 40 (5) ◽  
pp. 1260-1262 ◽  
Author(s):  
W C Ko ◽  
K W Yu ◽  
C Y Liu ◽  
C T Huang ◽  
H S Leu ◽  
...  

A total of 234 clinical isolates of Aeromonas, primarily A. hydrophila, were collected for the present study. Most were isolates from blood. By the agar dilution method, more than 90% of the Aeromonas strains were found to be susceptible to moxalactam, ceftazidime, cefepime, aztreonam, imipenem, amikacin, and fluoroquinolones, but they were more resistant to tetracycline, trimethoprim-sulfamethoxazole, some extended-spectrum cephalosporins, and aminoglycosides than strains from the United States and Australia.


mSphere ◽  
2020 ◽  
Vol 5 (5) ◽  
Author(s):  
Shivdeep Singh Hayer ◽  
Seunghyun Lim ◽  
Samuel Hong ◽  
Ehud Elnekave ◽  
Timothy Johnson ◽  
...  

ABSTRACT Fluoroquinolones and cephalosporins are critically important antimicrobial classes for both human and veterinary medicine. We previously found a drastic increase in enrofloxacin resistance in clinical Escherichia coli isolates collected from diseased pigs from the United States over 10 years (2006 to 2016). However, the genetic determinants responsible for this increase have yet to be determined. The aim of the present study was to identify and characterize the genetic basis of resistance against fluoroquinolones (enrofloxacin) and extended-spectrum cephalosporins (ceftiofur) in swine E. coli isolates using whole-genome sequencing (WGS). blaCMY-2 (carried by IncA/C2, IncI1, and IncI2 plasmids), blaCTX-M (carried by IncF, IncHI2, and IncN plasmids), and blaSHV-12 (carried by IncHI2 plasmids) genes were present in 87 (82.1%), 19 (17.9%), and 3 (2.83%) of the 106 ceftiofur-resistant isolates, respectively. Of the 110 enrofloxacin-resistant isolates, 90 (81.8%) had chromosomal mutations in gyrA, gyrB, parA, and parC genes. Plasmid-mediated quinolone resistance genes [qnrB77, qnrB2, qnrS1, qnrS2, and aac-(6)-lb′-cr] borne on ColE, IncQ2, IncN, IncF, and IncHI2 plasmids were present in 24 (21.8%) of the enrofloxacin-resistant isolates. Virulent IncF plasmids present in swine E. coli isolates were highly similar to epidemic plasmids identified globally. High-risk E. coli clones, such as ST744, ST457, ST131, ST69, ST10, ST73, ST410, ST12, ST127, ST167, ST58, ST88, ST617, ST23, etc., were also found in the U.S. swine population. Additionally, the colistin resistance gene (mcr-9) was present in several isolates. This study adds valuable information regarding resistance to critical antimicrobials with implications for both animal and human health. IMPORTANCE Understanding the genetic mechanisms conferring resistance is critical to design informed control and preventive measures, particularly when involving critically important antimicrobial classes such as extended-spectrum cephalosporins and fluoroquinolones. The genetic determinants of extended-spectrum cephalosporin and fluoroquinolone resistance were highly diverse, with multiple plasmids, insertion sequences, and genes playing key roles in mediating resistance in swine Escherichia coli. Plasmids assembled in this study are known to be disseminated globally in both human and animal populations and environmental samples, and E. coli in pigs might be part of a global reservoir of key antimicrobial resistance (AMR) elements. Virulent plasmids found in this study have been shown to confer fitness advantages to pathogenic E. coli strains. The presence of international, high-risk zoonotic clones provides worrisome evidence that resistance in swine isolates may have indirect public health implications, and the swine population as a reservoir for these high-risk clones should be continuously monitored.


2017 ◽  
Vol 4 (3) ◽  
Author(s):  
Nenad Macesic ◽  
Daniel Green ◽  
Zheng Wang ◽  
Sean B. Sullivan ◽  
Kevin Shim ◽  
...  

Abstract The spread of mcr-1 in the United States remains poorly defined. mcr-1-producing Escherichia coli that also carried blaSHV-12 was detected in a hospitalized patient. No additional cases were identified during screening of 801 Gram-negative isolates. Genomic sequencing identified an IncX4 mcr-1- harboring plasmid and ST117 clonal background associated with avian pathogenic E coli.


1976 ◽  
Vol 39 (3) ◽  
pp. 175-178 ◽  
Author(s):  
J. M. GOEPFERT

Nine hundred fifty-five samples of raw ground beef obtained from supermarkets throughout the United States were examined for coliforms, Escherichia coli, and Aerobic Plate Count (APC). The results were compared with existent standards for E. coli in raw meat in New York and Oregon. Lack of homogenious distribution of E. coli in fresh ground beef was demonstrated. Observations were made that indicate that a 2 day-two step procedure will detect the same number of E. coli as the more time consuming four step MPN procedure 98% of the time. A comparison of the APC obtained by incubating plates at 20 C and 35 C showed there to be an average 10-fold difference with the 20 C incubation always higher. Questions are raised about the necessity of microbial standards for raw meat and the validity of incorporating E. coli in such standards.


2007 ◽  
Vol 190 (3) ◽  
pp. 1054-1063 ◽  
Author(s):  
Cheryl-Lynn Y. Ong ◽  
Glen C. Ulett ◽  
Amanda N. Mabbett ◽  
Scott A. Beatson ◽  
Richard I. Webb ◽  
...  

ABSTRACT Catheter-associated urinary tract infection (CAUTI) is the most common nosocomial infection in the United States. Uropathogenic Escherichia coli (UPEC), the most common cause of CAUTI, can form biofilms on indwelling catheters. Here, we identify and characterize novel factors that affect biofilm formation by UPEC strains that cause CAUTI. Sixty-five CAUTI UPEC isolates were characterized for phenotypic markers of urovirulence, including agglutination and biofilm formation. One isolate, E. coli MS2027, was uniquely proficient at biofilm growth despite the absence of adhesins known to promote this phenotype. Mini-Tn5 mutagenesis of E. coli MS2027 identified several mutants with altered biofilm growth. Mutants containing insertions in genes involved in O antigen synthesis (rmlC and manB) and capsule synthesis (kpsM) possessed enhanced biofilm phenotypes. Three independent mutants deficient in biofilm growth contained an insertion in a gene locus homologous to the type 3 chaperone-usher class fimbrial genes of Klebsiella pneumoniae. These type 3 fimbrial genes (mrkABCDF), which were located on a conjugative plasmid, were cloned from E. coli MS2027 and could complement the biofilm-deficient transconjugants when reintroduced on a plasmid. Primers targeting the mrkB chaperone-encoding gene revealed its presence in CAUTI strains of Citrobacter koseri, Citrobacter freundii, Klebsiella pneumoniae, and Klebsiella oxytoca. All of these mrkB-positive strains caused type 3 fimbria-specific agglutination of tannic acid-treated red blood cells. This is the first description of type 3 fimbriae in E. coli, C. koseri, and C. freundii. Our data suggest that type 3 fimbriae may contribute to biofilm formation by different gram-negative nosocomial pathogens.


2016 ◽  
Vol 60 (7) ◽  
pp. 4073-4081 ◽  
Author(s):  
Kalyan D. Chavda ◽  
Liang Chen ◽  
Michael R. Jacobs ◽  
Robert A. Bonomo ◽  
Barry N. Kreiswirth

ABSTRACTThe emergence and spread ofKlebsiella pneumoniaecarbapenemase (KPC) amongEnterobacteriaceaepresents a major public health threat to the world. Although not as common as inK. pneumoniae, KPC is also found inEscherichia colistrains. Here, we genetically characterized 9 carbapenem-resistantE. colistrains isolated from six hospitals in the United States and completely sequenced theirblaKPC-harboring plasmids. The nine strains were isolated from different geographical locations and belonged to 8 differentE. colisequence types. SevenblaKPC-harboring plasmids belonged to four different known incompatibility groups (IncN, -FIA, -FIIK2, and -FIIK1) and ranged in size from ∼16 kb to ∼241 kb. In this analysis, we also identified two plasmids that have novel replicons: (i) pBK28610, which is similar to p34978-3 with an insertion of Tn4401b, and (ii) pBK31611, which does not have an apparent homologue in the GenBank database. Moreover, we report the emergence of a pKP048-like plasmid, pBK34397, inE. coliin the United States. Meanwhile, we also found examples of interspecies spread ofblaKPCplasmids, as pBK34592 is identical to pBK30683, isolated fromK. pneumoniae. In addition, we discovered examples of acquisition (pBK32602 acquired an ∼46-kb fragment including a novel replication gene, along with Tn4401band other resistance genes) and/or loss (pKpQIL-Ec has a 14.5-kb deletion compared to pKpQIL-10 and pBK33689) of DNA, demonstrating the plasticity of these plasmids and their rapid evolution in the clinic. Overall, our study shows that the spread ofblaKPC-producingE. coliis largely due to horizontal transfer ofblaKPC-harboring plasmids and related mobile elements into diverse genetic backgrounds.


2004 ◽  
Vol 48 (4) ◽  
pp. 1295-1299 ◽  
Author(s):  
Minggui Wang ◽  
Daniel F. Sahm ◽  
George A. Jacoby ◽  
David C. Hooper

ABSTRACT Although quinolone resistance commonly results from chromosomal mutation, recent studies indicate that such resistance can also be transferred on plasmids carrying the gene responsible, qnr. One hundred ten ciprofloxacin-resistant clinical isolates of Klebsiella pneumoniae and Escherichia coli from the United States were screened for the qnr gene by PCR and Southern hybridization of plasmid DNA. Conjugation experiments were done with azide-resistant E. coli J53 as the recipient and selection with azide and sulfonamide, a resistance frequently linked to qnr. EcoRI and BamHI digests of qnr-hybridizing plasmids were subjected to electrophoresis on agarose gels and probed with qnr by Southern hybridization. qnr was detected in 8 (11.1%) of 72 K. pneumoniae strains. These eight positive strains were from six states in the United States. qnr was not found in any of the 38 E. coli strains tested. Quinolone resistance was transferred from seven of the eight probe-positive strains. Transconjugants with qnr-hybridizing plasmids had 32-fold increases in ciprofloxacin MICs relative to E. coli J53. For all eight strains, the sequence of qnr was identical to that originally reported. By size and restriction digests, four plasmids were related to the first-reported plasmid, pMG252, and three were different. Five new qnr plasmids encoded FOX-5 β-lactamase, as did pMG252, but two others produced SHV-7 extended-spectrum β-lactamase. Transferable plasmid-mediated quinolone resistance associated with qnr is now widely distributed in quinolone-resistant clinical strains of K. pneumoniae in the United States. Plasmid-determined quinolone resistance contributes to the increasing quinolone resistance of K. pneumoniae isolates and to the linkage previously observed between resistance to quinolones and the latest β-lactam antibiotics.


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