Comparison of Plasmids Coharboring 16S rRNA Methylase and Extended-Spectrum β-Lactamase Genes among Escherichia coli Isolates from Pets and Poultry

2013 ◽  
Vol 76 (12) ◽  
pp. 2018-2023 ◽  
Author(s):  
XIAO-PING LIAO ◽  
BAO-TAO LIU ◽  
QIU-E YANG ◽  
JIAN SUN ◽  
LIANG LI ◽  
...  

A total of 247 Escherichia coli isolates (148 from diseased or dead poultry and 99 from diseased pets in the People's Republic of China) were screened for extended-spectrum β-lactamase (ESBL) determinants by PCR and sequencing. Then, 16S rRNA methylase genes were detected among ESBL-producing isolates. Clonal relatedness of the E. coli isolates was examined by pulsed-field gel electrophoresis. Conjugation experiments were performed to investigate the association of 16S rRNA methylases and ESBLs, and plasmid contents were also characterized. Among 247 E. coli isolates, 74 (29.96%) isolates were positive for blaCTX-M genes, 42 from pets (12 from cats and 30 from dogs) and 32 from poultry (12 from chickens and 20 from ducks). The most common CTX-M type in isolates from pets was blaCTX-M-14, whereas blaCTX-M-27 was the most common for poultry. rmtB was dectected in 39 of the 74 blaCTX-M-positive isolates, 18 from pets and 21 from poultry. One strain from a pet was found to harbor blaCTX-M-14, blaCTX-M-15, and rmtB. blaCTX-M and rmtB were found to be colocated on the same transferable plasmid in 16 isolates. These genes were on the same or similar plasmids (eight F2:A-:B- and two IncN) in isolates from ducks, whereas they were colocated on the similar F2:A-:B- or similar F33:A-:B- plasmids in isolates of pets origin. In conclusion, similar F2:A-:B-plasmids and similar F33:A-:B- plasmids are responsible for the dissemination of both rmtB and blaCTX-M genes in E. coli isolates from poultry and pets, respectively.

2006 ◽  
Vol 50 (2) ◽  
pp. 785-787 ◽  
Author(s):  
Alexandra Eisner ◽  
Elizabeth J. Fagan ◽  
Gebhard Feierl ◽  
Harald H. Kessler ◽  
Egon Marth ◽  
...  

ABSTRACT Among 149 extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae isolates collected from patients in southeast Austria from 1998 to 2004, 38 Escherichia coli isolates and 11 Klebsiella spp. were CTX-M producers. The proportion of CTX-M-producers among all ESBL producers rose from 0% in 1998 to 58% in 2004. In general, CTX-M-producers had heterogeneous pulsed-field gel electrophoresis patterns, but one E. coli isolate was identical to a United Kingdom epidemic CTX-M-15-producing strain, although no epidemiological link with the United Kingdom was apparent.


2005 ◽  
Vol 49 (3) ◽  
pp. 1262-1264 ◽  
Author(s):  
Laura Briñas ◽  
Miguel Angel Moreno ◽  
Tirushet Teshager ◽  
Yolanda Sáenz ◽  
María Concepción Porrero ◽  
...  

ABSTRACT Genes encoding CTX-M-14, CTX-M-9, CTX-M-1, CTX-M-32, SHV-12, TEM-52, or CMY-2 β-lactamases were detected in 21 Escherichia coli strains recovered during 2003 from sick animals (11 of 459 [2.4%] strains) and healthy animals (10 of 158 [6.3%] strains) in Spain. Twelve of these strains harbored bla CTX-M genes and showed unrelated pulsed-field gel electrophoresis patterns.


2017 ◽  
Vol 119 (7) ◽  
pp. 1633-1647
Author(s):  
Kaotar Nayme ◽  
Abouddihaj Barguigua ◽  
Brahim Bouchrif ◽  
Idrissa Diawara ◽  
Fatima El Otmani ◽  
...  

Purpose The purpose of this paper is to assess the occurrence of the extended-spectrum β-lactamases (ESBLs) and AmpC β-lactamase genes in 144 Escherichia coli isolates recovered from 160 vegetable salad samples. Design/methodology/approach Among the 144 E. coli isolates recovered from 160 vegetable salads, 17 (12 percent) ceftazidime-resistant isolates were screened for ESBL production with the double disk-diffusion test. The ESBL-producing isolates were characterized for antimicrobial resistance, the presence of virulence genes and plasmid-mediated quinolone resistance (PMQR) determinants. The isolates were also subjected to phylogenetic group typing. The existence of plasmid AmpC genes and mutations in the regulatory region of the chromosomal AmpC gene was assessed using polymerase chain reaction (PCR) and sequencing. All β-lactamase isolates were further characterized by pulsed-field gel electrophoresis to determine the genetic relatedness. Findings Overall, 17 (12 percent) of the 144 E. coli isolates studied were ceftazidime resistant. Among the 17 isolates, 13 (77 percent) were multidrug resistant and four (23.5 percent) were ESBL producers. The bla CTX-M14 was the only gene detected. Of the 12 AmpC-producing isolates, three (18 percent) harbored plasmid-encoded AmpC and sequencing analysis of the chromosomal AmpC genes revealed mutations in the promoter/attenuator region. PMQR determinants were detected in 9 (52 percent) isolates. A was the most prevalent phylogenetic group (56 percent), followed by groups B1 (31 percent), D (6 percent), and B2 (6 percent). PCR showed that six (50 percent) ESBL/AmpC-producing E. coli isolates carried one and/or two virulence genes. Pulsed-field gel electrophoresis showed no epidemiological relationship between these isolates. Originality/value This study places vegetable salads within the spectrum of ecological niches that may be vehicles for antibiotic-resistant bacteria/genes with clinical interest and these findings are worthy of attention as their spread to humans by ingestion cannot be dismissed.


2015 ◽  
Vol 59 (10) ◽  
pp. 6611-6614 ◽  
Author(s):  
Thi Mai Huong Bui ◽  
Itaru Hirai ◽  
Shuhei Ueda ◽  
Thi Kim Ngan Bui ◽  
Kouta Hamamoto ◽  
...  

ABSTRACTHealthy carriage of CTX-M-type extended-spectrum β-lactamase (ESBL)-producingEscherichia coliwas examined by thrice collecting fecal samples from the same 199 healthy Vietnamese subjects every 6 months. Using pulsed-field gel electrophoresis (PFGE), identical PFGE patterns throughout the three samplings were not observed, although prevalence ofE. coliin the subjects was around 50% in the three samplings. Our results suggested a short carriage period of the CTX-M-type ESBL-producingE. coliin healthy Vietnamese subjects.


2009 ◽  
Vol 53 (11) ◽  
pp. 4733-4739 ◽  
Author(s):  
Hanna E. Sidjabat ◽  
David L. Paterson ◽  
Jennifer M. Adams-Haduch ◽  
Lindsay Ewan ◽  
Anthony W. Pasculle ◽  
...  

ABSTRACT A combination of phenotypic and genotypic methods was used to investigate 70 unique Escherichia coli clinical isolates identified as producing extended-spectrum β-lactamases (ESBLs) at a medical center in Pittsburgh, PA, between 2007 and 2008. Fifty-seven isolates (81%) produced CTX-M-type ESBLs, among which CTX-M-15 was predominant (n = 46). Isolates producing CTX-M-2, -9, -14, and -65 were also identified. One CTX-M-producing isolate coproduced CMY-2 cephalosporinase. Ten isolates (14%) produced SHV-type ESBLs, either SHV-5 or SHV-7. Two isolates produced only CMY-2 or -32. Pulsed-field gel electrophoresis revealed the presence of two major clusters of CTX-M-15-producing E. coli isolates, one in phylotype B2 (n = 15) and the other in phylotype A (n = 19). Of four phylotype B2 isolates that were able to transfer the bla CTX-M-15-carrying plasmids, three showed fingerprints related (>60%) to those of plasmids from phylotype A isolates. In phylotype B2, all CTX-M-15-producing isolates, as well as three isolates producing CTX-M-14, two producing SHV-5, and one producing SHV-7, belonged to the international epidemic clone defined by serotype O25:H4 and sequence type 131. The plasmids from eight of nine CTX-M-15-producing E. coli isolates of phylotype A that were examined were highly related to each other and were also found in two isolates belonging to phylotype D, suggesting horizontal transfer of this bla CTX-M-15-carrying plasmid between phylotypes. Our findings underscore the need to further investigate the epidemiology and virulence of CTX-M-producing E. coli in the United States.


2006 ◽  
Vol 50 (2) ◽  
pp. 534-541 ◽  
Author(s):  
Jun-ichi Wachino ◽  
Hiroshi Kurokawa ◽  
Satowa Suzuki ◽  
Kunikazu Yamane ◽  
Naohiro Shibata ◽  
...  

ABSTRACT Nine Escherichia coli and 5 Klebsiella pneumoniae clinical isolates resistant to various cephalosporins and cephamycins were identified in a Japanese general hospital between 1995 and 1997. All nine E. coli isolates and one K. pneumoniae isolate carried bla CMY-9, while the other four K. pneumoniae isolates harbored a variant of bla CMY-9, namely, bla CMY-19. The pulsed-field gel electrophoresis patterns of the nine CMY-9-producing E. coli isolates were almost identical, suggesting their clonal relatedness, while those of the five K. pneumoniae isolates were divergent. Plasmid profiles, Southern hybridization, and conjugation assays revealed that the genes for the CMY-9 and the CMY-19 β-lactamases were located on very similar conjugative plasmids in E. coli and K. pneumoniae. The genetic environment of bla CMY-19 was identical to that of bla CMY-9. A single amino acid substitution, I292S, adjacent to the H-10 helix region was observed between CMY-9 and CMY-19. This substitution was suggested to be responsible for the expansion of the hydrolyzing activity against several broad-spectrum cephalosporins, and this finding was consistent with the kinetic parameters determined with purified enzymes. These findings suggest that the bla CMY-19 genes found in the four K. pneumoniae isolates might have originated from bla CMY-9 gene following a point mutation and dispersed among genetically different K. pneumoniae isolates via a large transferable plasmid.


2016 ◽  
Vol 10 (01) ◽  
pp. 43-52 ◽  
Author(s):  
Rafael Vignoli ◽  
Virginia García-Fulgueiras ◽  
Nicolás F Cordeiro ◽  
Inés Bado ◽  
Verónica Seija ◽  
...  

Introduction: To characterize extended-spectrum β-lactamases (ESBLs) and plasmid-mediated quinolone resistance (PMQR) genes in Escherichia coli isolates obtained from extra-intestinal samples in three Uruguayan hospitals. Methodology: Fifty-five ESBL-producing E. coli isolates were studied. Virulence genes, ESBLs, and PMQR genes were detected by polymerase chain reaction.  ESBL-producing isolates were compared by pulsed-field gel electrophoresis. Multi-locus sequence typing was also performed on 13 selected isolates. Results: Thirty-seven isolates harbored blaCTX-M-15 (67.3%), eight blaCTX-M-2 (14.6%), five blaCTX-M-14 (9.1%), three carried both blaCTX-M-2 and blaCTX-M-14, one blaCTX-M-9, and one blaCTX-M-8.  Among the CTX-M-15 producers, 92% belonged to sequence types ST131 and ST405, and carried aac(6’)Ib-cr as well. Isolates harboring blaCTX-M-2, blaCTX-M-14, blaCTX-M-9, or blaCTX-M-8 were found to be genetically unrelated. Conclusions: The successful dissemination of CTX-M-15-producing E.coli isolates seems to be linked to the spreading of high-risk clones and horizontal gene transfer. A trade-off between carrying more antibiotic resistance and less virulence-related genes could partially account for the evolutionary advantages featured by successful clones.


2005 ◽  
Vol 49 (11) ◽  
pp. 4754-4756 ◽  
Author(s):  
María-Rosario Baquero ◽  
Juan Carlos Galán ◽  
María del Carmen Turrientes ◽  
Rafael Cantón ◽  
Teresa M. Coque ◽  
...  

ABSTRACT Hypermutable (mutation frequency [f], ≥4 × 10−8) Escherichia coli strains were more frequently found (43%) in a collection of 89 extended-spectrum β-lactamase (ESBL)-producing isolates from different patients (77 pulsed-field gel electrophoresis clones, 12 ESBL types) than in non-ESBL E. coli (26%) strains (P = 0.03). Among urinary tract isolates, the frequency of hypermutation was 40% in ESBL versus 26% in non-ESBL isolates (P = 0.03).


2015 ◽  
Vol 78 (5) ◽  
pp. 1018-1023 ◽  
Author(s):  
MEILI XI ◽  
QIAN WU ◽  
XIN WANG ◽  
BAOWEI YANG ◽  
XIAODONG XIA ◽  
...  

Extended-spectrum β-lactamase (ESBL)–producing Escherichia coli strains have been reported worldwide; however, the incidence and characterization of foodborne ESBL-producing E. coli strains have been rarely reported in the People's Republic of China. Among a collection of 659 E. coli isolates recovered from retail foods in Shaanxi Province, People's Republic of China, 223 cefoxitin-resistant and/or cefoperazone-resistant isolates were screened for ESBL production with the double disk diffusion test. The ESBL-producing isolates were characterized for antimicrobial resistance and the presence of blaTEM, blaSHV, and blaCTX-M genes. Isolates with blaCTX-M were further classified by PCR as having blaCTX-M-1, blaCTX-M-2, blaCTX-M-8, blaCTX-M-9, or blaCTX-M-25. One hundred forty-seven isolates were identified as ESBL positive. PCR detection revealed that 146 isolates (99.3%) contained the blaCTX-M gene. Among these isolates, 42 (28.8%) were positive for the enzyme CTX-M-1, 5 (3.4%) for CTX-M-2, and 99 (67.8%) for CTX-M-9. No CTX-M-8 and CTX-M-25 were found in this study. One hundred fifteen isolates (78.2%) were positive for the blaTEM gene, but blaSHV was not detected. Among the 147 ESBL-producing E. coli isolates, 75 (51.0%), 35 (23.8%), and 4 (2.7%) isolates were positive for blaTEM and blaCTX-M-9, blaTEM and blaCTX-M-1, and blaTEM and blaCTX-M-2, respectively. All of the 147 ESBL-producing isolates were resistant to three or more non–β-lactam antibiotics. This study provides evidence that foodborne E. coli can harbor ESBL-encoding genes. Thus, food could be a vehicle for the dissemination of ESBL-producing E. coli strains, a situation that requires surveillance and appropriate management strategies.


1999 ◽  
Vol 122 (2) ◽  
pp. 337-341 ◽  
Author(s):  
M. AKIBA ◽  
T. MASUDA ◽  
T. SAMESHIMA ◽  
K. KATSUDA ◽  
M. NAKAZAWA

A total of 77 Escherichia coli O157[ratio ]H7 (H−) isolates from cattle in Japan were investigated by molecular biological methods. Most of these isolates (43 isolates) possessed the stx2 gene, but not stx1. Fifteen bacteriophage types and 50 pulsed-field gel electrophoresis (PFGE) profiles were observed. One isolate was indistinguishable from the human outbreak strain by these methods. This indicates that cattle must be considered as a possible source of human E. coli O157[ratio ]H7 infection in Japan.


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