Conjugative Plasmid-Mediated Extended Spectrum Cephalosporin Resistance in Genetically Diverse Escherichia coli from a Chicken Slaughterhouse

ESC-resistant E. coli isolates were collected from broiler chickens, a slaughterhouse, and retail meat to assess their dispersion and their involvement in cross-contamination. ESBL-/AmpC-producing E. coli were isolated during the slaughter process of all six investigated chicken flocks from scalding, feather removal, first conveyor, evisceration, second washing, third conveyor, and third washing areas, and from handling workers in the slaughterhouse. ESC-resistant E. coli isolates with the same pulsed-field gel electrophoresis type were found in the same site (scalding) on different sampling days. ESBL/AmpC-producing E. coli isolates were absent in the lairage area in the slaughterhouse, but present in the retail markets in 36.8% (7/19) of the chicken flocks. The blaCTX-M genes and blaCMY-2 were conjugated to recipient E. coli J53 in 67.5% (27/40) and 56.1% (23/41) of ESBL-producing and AmpC-producing E. coli isolates, respectively. The presence of the same conjugative plasmids was found in genetic diversity ESC-resistant E. coli colonies collected on different sampling days. Our study emphasizes that cross-contamination of ESBL/AmpC-producing E. coli in slaughterhouse has a crucial impact on the occurrence of ESC resistance in retail chicken meat.

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Molecular typing of Escherichia coli O157[ratio ]H7 (H−) isolates from cattle in Japan

Epidemiology and Infection ◽

10.1017/s0950268899002198 ◽

1999 ◽

Vol 122 (2) ◽

pp. 337-341 ◽

Cited By ~ 16

Author(s):

M. AKIBA ◽

T. MASUDA ◽

T. SAMESHIMA ◽

K. KATSUDA ◽

M. NAKAZAWA

Keyword(s):

Escherichia Coli ◽

Molecular Typing ◽

Gel Electrophoresis ◽

Pulsed Field Gel Electrophoresis ◽

Escherichia Coli O157 ◽

Outbreak Strain ◽

Pulsed Field ◽

E Coli ◽

Biological Methods ◽

Pfge Profiles

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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Isolation and plasmid characterisation of Salmonella enterica serovar Albany harbouring mcr-5 from retail chicken meat in Japan

FEMS Microbiology Letters ◽

10.1093/femsle/fnaa127 ◽

2020 ◽

Vol 367 (15) ◽

Author(s):

Yuki Wakabayashi ◽

Tsuyoshi Sekizuka ◽

Takahiro Yamaguchi ◽

Akira Fukuda ◽

Masato Suzuki ◽

...

Keyword(s):

Meat Products ◽

Asian Countries ◽

Colistin Resistance ◽

Single Nucleotide Variants ◽

Single Nucleotide ◽

E Coli ◽

Retail Meat ◽

Paratyphi B ◽

Retail Chicken Meat ◽

Plasmid Backbone

ABSTRACT The emergence of plasmid-mediated colistin resistance genes (mcr), which is occurring in numerous countries, is a worldwide concern, primarily because colistin is a last-resort antibiotic. Compared to E. coli, prevalence of mcr genes in Salmonella is unclear in Japan. Here we screened for mcr-1–5 genes in our collection of Salmonella strains isolated from retail meat products collected in Japan from 2012 through 2016. We found that Salmonella Albany strain 27A-368 encodes mcr-5 and that mcr genes were undetectable among the remaining 202 isolates. The resistance plasmid p27A-368 was transferred by conjugation to S. Infantis and was stably retained as a transconjugant. Whole-genome sequencing revealed that mcr-5 resided on a 115 kb plasmid (p27A-368). The plasmid backbone of p27A-368 is more similar to that of pCOV27, an ESBL-encoding plasmid recovered from avian pathogenic E. coli, rather than pSE13-SA01718 of S. Paratyphi B that encodes mcr-5. Further, mcr-5 is located on a transposon, and its sequence is similar to that of pSE13-SA01718. A phylogenetic tree based on single nucleotide variants implies a relationship between 27A-368 and S. Albany isolated in Southeast Asian countries.

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Evaluation of Logistic Processing To Reduce Cross-Contamination of Commercial Broiler Carcasses with Campylobacter spp.

Journal of Food Protection ◽

10.4315/0362-028x-70.11.2549 ◽

2007 ◽

Vol 70 (11) ◽

pp. 2549-2554 ◽

Cited By ~ 18

Author(s):

LAKSHMI-PRASANNA POTTURI-VENKATA ◽

STEFFEN BACKERT ◽

SERGIO L. VIEIRA ◽

OMAR A. OYARZABAL

Keyword(s):

Gel Electrophoresis ◽

Food Production ◽

Pulsed Field Gel Electrophoresis ◽

Cross Contamination ◽

Large Problem ◽

Pulsed Field ◽

Before And After ◽

Commercial Broiler ◽

Simple Logistic ◽

Campylobacter Spp

Cross-contamination of broiler carcasses with Campylobacter is a large problem in food production. Here, we investigated whether the contamination of broilers carcasses from Campylobacter-negative flocks can be avoided by logistic scheduling during processing. For this purpose, fecal samples were collected from several commercial broiler flocks and enumerated for Campylobacter spp. Based on enumeration results, flocks were categorized as Campylobacter negative or Campylobacter positive. The schedule of processing included the testing of Campylobacter-positive flocks before or after the testing of Campylobacter-negative flocks. During processing, flocks were also sampled for Campylobacter spp. before and after chilling. Campylobacter strains were identified with multiplex PCR and analyzed for relatedness with pulsed-field gel electrophoresis. Our results show that Campylobacter-negative flocks were indeed contaminated with Campylobacter strains originating from previously processed Campylobacter-positive flocks. Campylobacter isolates collected from carcasses originating from different farms processed on the same day showed similar pulsed-field gel electrophoresis patterns, confirming cross-contamination. These findings suggest that a simple logistic processing schedule can preserve the Campylobacter-negative status of broiler carcasses and result in products with enhanced food safety.

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Shedding of Escherichia coli O157:H7 in Dairy Cattle Housed in a Confined Environment following Waterborne Inoculation

Applied and Environmental Microbiology ◽

10.1128/aem.68.4.1947-1954.2002 ◽

2002 ◽

Vol 68 (4) ◽

pp. 1947-1954 ◽

Cited By ~ 43

Author(s):

J. A. Shere ◽

C. W. Kaspar ◽

K. J. Bartlett ◽

S. E. Linden ◽

B. Norell ◽

...

Keyword(s):

Escherichia Coli ◽

Water Treatment ◽

Gel Electrophoresis ◽

Pulsed Field Gel Electrophoresis ◽

Escherichia Coli O157 ◽

E Coli ◽

Bull Calves ◽

Genomic Dnas ◽

Confined Environment ◽

Effective Water

ABSTRACT A study of Escherichia coli O157:H7 transmission and shedding was conducted with bull calves housed in individual pens within a confined environment. For comparative purposes, the numbers and duration of E. coli O157:H7 shedding in naturally infected calves were monitored after a single purchased calf (calf 156) tested positive prior to inoculation. During the next 8 days, the calves in adjacent pens and a pen directly across a walkway from calf 156 began to shed this serotype O157:H7 strain. Five of the eight calves in this room shed this O157:H7 strain at some time during the following 8 weeks. The numbers of E. coli O157:H7 isolates shed in these calves varied from 60 to 105 CFU/g of feces, and the duration of shedding ranged from 17 to >31 days. The genomic DNAs from isolates recovered from these calves were indistinguishable when compared by using XbaI digestion and pulsed-field gel electrophoresis. Inoculation of calves with 1 liter of water containing ca. 103 to 104 CFU of E. coli O157:H7/ml resulted in shedding in 10 of 12 calves (trial 1, 4 of 4 calves; trial 2, 6 of 8 calves). The inoculated calves shed the inoculation strain (FRIK 1275) as early as 24 h after administration. The duration of shedding varied from 18 to >43 days at levels from 102 to 106 CFU/g of feces. The numbers of doses necessary to initiate shedding varied among calves, and two calves in trial 2 never shed FRIK 1275 after four doses (ca. 106 CFU per dose). Results from this study confirm previous reports of animal-to-animal and waterborne dissemination of E. coli O157:H7 and highlight the need for an effective water treatment to reduce the spread of this pathogen in cattle.

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Endoscopic Retrograde Cholangiopancreatography–Associated AmpCEscherichia coliOutbreak

Infection Control and Hospital Epidemiology ◽

10.1017/ice.2015.66 ◽

2015 ◽

Vol 36 (6) ◽

pp. 634-642 ◽

Cited By ~ 119

Author(s):

Kristen A. Wendorf ◽

Meagan Kay ◽

Christopher Baliga ◽

Scott J. Weissman ◽

Michael Gluck ◽

...

Keyword(s):

Infection Control ◽

Endoscopic Retrograde Cholangiopancreatography ◽

Gel Electrophoresis ◽

Gene Sequencing ◽

Pulsed Field Gel Electrophoresis ◽

Control Measures ◽

Biliary Disease ◽

Endoscopic Retrograde ◽

Pulsed Field ◽

E Coli

BACKGROUNDWe identified an outbreak of AmpC–producingEscherichia coliinfections resistant to third-generation cephalosporins and carbapenems (CR) among 7 patients who had undergone endoscopic retrograde cholangiopancreatography at hospital A during November 2012–August 2013. Gene sequencing revealed a shared novel mutation in ablaCMYgene and a distinctivefumC/ fimHtyping profile.OBJECTIVETo determine the extent and epidemiologic characteristics of the outbreak, identify potential sources of transmission, design and implement infection control measures, and determine the association between the CRE. coliand AmpCE. colicirculating at hospital A.METHODSWe reviewed laboratory, medical, and endoscopy reports, and endoscope reprocessing procedures. We obtained cultures from endoscopes after reprocessing as well as environmental samples and conducted pulsed-field gel electrophoresis and gene sequencing on phenotypic AmpC isolates from patients and endoscopes. Cases were those infected with phenotypic AmpC isolates (both carbapenem-susceptible and CR) and identicalblaCMY-2,fumC, andfimHalleles or related pulsed-field gel electrophoresis patterns.RESULTSThirty-five of 49 AmpCE. colitested met the case definition, including all CR isolates. All cases had complicated biliary disease and had undergone at least 1 endoscopic retrograde cholangiopancreatography at hospital A. Mortality at 30 days was 16% for all patients and 56% for CR patients. Two of 8 reprocessed endoscopic retrograde cholangiopancreatography scopes harbored AmpC that matched case isolates by pulsed-field gel electrophoresis. Environmental cultures were negative. No breaches in infection control were identified. Endoscopic reprocessing exceeded manufacturer’s recommended cleaning guidelines.CONCLUSIONRecommended reprocessing guidelines are not sufficient.Infect Control Hosp Epidemiol2015;00(0): 1–9

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Fresh Steam-Flaked Corn in Cattle Feedlots Is an Important Site for Fecal Coliform Contamination by House Flies†

Journal of Food Protection ◽

10.4315/0362-028x.jfp-14-429 ◽

2015 ◽

Vol 78 (3) ◽

pp. 567-572 ◽

Cited By ~ 7

Author(s):

ANURADHA GHOSH ◽

LUDEK ZUREK

Keyword(s):

Gel Electrophoresis ◽

Fecal Coliform ◽

Bacterial Contamination ◽

Pulsed Field Gel Electrophoresis ◽

Fecal Coliforms ◽

House Flies ◽

E Coli ◽

Klebsiella Spp ◽

Important Site ◽

Cattle Feedlots

House flies are a common pest at food animal facilities, including cattle feedlots. Previously, house flies were shown to play an important role in the ecology of Escherichia coli O157:H7; house flies in cattle feedlots carried this zoonotic pathogen and were able to contaminate cattle through direct contact and/or by contamination of drinking water and feed. Because house flies aggregate in large numbers on fresh (≤6 h) steam-flaked corn (FSFC) used in cattle feed, the aim of this study was to assess FSFC in a cattle feedlot as a potentially important site of fecal coliform contamination by house flies. House flies and FSFC samples were collected, homogenized, and processed for culturing of fecal coliforms on membrane fecal coliform agar. Selected isolates were identified by 16S rRNA gene sequencing, and representative isolates from each phylogenetic group were genotyped by pulsed-field gel electrophoresis. Fecal coliforms were undetectable in FSFC shortly (0 h) after flaking; however, in summer, after 4 to 6 h, the concentrations of fecal coliforms ranged from 1.9 × 103 to 3.7 × 104 CFU/g FSFC (mean, 1.1 ± 3.0 × 104 CFU/g). House flies from FSFC carried between 7.6 × 102 and 4.1 × 106 CFU of fecal coliforms per fly (mean, 6.0 ± 2.3 × 105 CFU per fly). Fecal coliforms were represented by E. coli (85.1%), Klebsiella spp. (10.6%), and Citrobacter spp. (4.3%). Pulsed-field gel electrophoresis demonstrated clonal matches of E. coli and Klebsiella spp. between house flies and FSFC. In contrast, in winter and in the absence of house flies, the contamination of corn by fecal coliforms was significantly (~10-fold) lower. These results indicate that FSFC is an important site for bacterial contamination by flies and possible exchange of E. coli and other bacteria among house flies. Further research is needed to evaluate the potential use of screens or blowers to limit the access of house flies to FSFC and therefore their effectiveness in preventing bacterial contamination.

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Molecular Characterization of a Novel Plasmid-Encoded Cefotaximase (CTX-M-12) Found in Clinical Klebsiella pneumoniae Isolates from Kenya

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.45.7.2141-2143.2001 ◽

2001 ◽

Vol 45 (7) ◽

pp. 2141-2143 ◽

Cited By ~ 53

Author(s):

S. Kariuki ◽

J. E. Corkill ◽

G. Revathi ◽

R. Musoke ◽

C. A. Hart

Keyword(s):

Klebsiella Pneumoniae ◽

Gel Electrophoresis ◽

Direct Sequencing ◽

Hydrolytic Activity ◽

Pulsed Field Gel Electrophoresis ◽

Chromosomal Dna ◽

National Hospital ◽

Cephalosporin Resistance ◽

M 12

ABSTRACT Nine Klebsiella pneumoniae isolates, six from blood and three from cerebrospinal fluid of newborn babies at Kenyatta National Hospital, Nairobi, Kenya, were analyzed for the mechanism of cephalosporin resistance. By using pulsed-field gel electrophoresis ofXbaI-digested chromosomal DNA, all the nine isolates were found to be clonal. PCR and direct sequencing revealed a novel extended-spectrum β-lactamase, which we designated CTX-M-12. It has a more potent hydrolytic activity against cefotaxime than against ceftazidime and a pI of 9.0 and is encoded on a large self-transferable ca. 160-kbp plasmid.

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A Mixture of Lactobacillus casei, Lactobacillus lactis, and Paenibacillus polymyxa Reduces Escherichia coli O157:H7 in Finishing Feedlot Cattle

Journal of Food Protection ◽

10.4315/0362-028x.jfp-13-433 ◽

2014 ◽

Vol 77 (5) ◽

pp. 738-744 ◽

Cited By ~ 7

Author(s):

KIM STANFORD ◽

SUSAN BACH ◽

JOHN BAAH ◽

TIM McALLISTER

Keyword(s):

Gel Electrophoresis ◽

Lactobacillus Casei ◽

Dry Matter ◽

Paenibacillus Polymyxa ◽

Pulsed Field Gel Electrophoresis ◽

Escherichia Coli O157 ◽

E Coli ◽

Growing Period ◽

Fecal Ph ◽

Lactobacillus Lactis

A direct-fed microbial (DFM) containing Paenibacillus polymyxa, Lactobacillus casei, and Lactobacillus lactis was fed to cattle (n = 120) to determine impacts on shedding and survival of Escherichia coli O157:H7 in feces. Cattle were individually penned and fed diets containing 0 (control), 4 × 107 CFU (DFM-4), 8 × 107 CFU (DFM-8), or 1.2 × 108 CFU (DFM-12) lactobacilli per kg of dietary dry matter over 84-day fall-winter growing and 140-day spring-summer finishing periods. Fecal grab samples were collected from cattle at 28-day intervals, E. coli O157:H7 was detected by immunomagnetic separation, and isolates were compared by pulsed-field gel electrophoresis. During the growing period, feces negative for E. coli O157 from each dietary treatment were inoculated with 105 CFU/g nalidixic acid–resistant E. coli O157:H7 and were incubated at 4 and 22°C for 11 weeks. Fecal pH and fecal dry matter were measured on days 0, 1, 3, and 7 and weekly thereafter, with E. coli O157:H7 enumerated through dilution plating. Treatment with DFMs did not affect survival of E. coli O157:H7 in feces or fecal pH (P > 0.05). Only one steer was positive for E. coli O157:H7 during the growing period, but during the finishing period, DFM-8 and DFM-12 reduced the prevalence of E. coli O157:H7 in feces (P < 0.05). Feeding DFMs also reduced the frequency of individual steers shedding E. coli O157:H7 during finishing (P < 0.05), with control steers shedding E. coli O157:H7 up to four times, whereas DFM-12 steers shed E. coli O157:H7 a maximum of twice. Treatment with DFMs influenced pulsed-field gel electrophoresis profiles; steers that were fed DFM-8 and DFM-12 shed more diverse subtypes of E. coli O157:H7 than did control or DFM-4 steers. Because a companion study found linear improvement in performance with increasing dosage of DFMs in the first 28 days of the growing period, targeted use of DFM-12 during this time and for the final 1 or 2 weeks prior to slaughter may optimize performance and reduce E. coli O157:H7 while minimizing feed costs.

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Long-Term Care Facilities Are Reservoirs for Antimicrobial-Resistant Sequence Type 131 Escherichia coli

Open Forum Infectious Diseases ◽

10.1093/ofid/ofv011 ◽

2015 ◽

Vol 2 (1) ◽

Cited By ~ 25

Author(s):

Mary J. Burgess ◽

James R. Johnson ◽

Stephen B. Porter ◽

Brian Johnston ◽

Connie Clabots ◽

...

Keyword(s):

Risk Factors ◽

Escherichia Coli ◽

Gel Electrophoresis ◽

Long Term Care ◽

Pulsed Field Gel Electrophoresis ◽

Sequence Type ◽

Term Care ◽

E Coli ◽

Care Facilities

Abstract Background. Emerging data implicate long-term care facilities (LTCFs) as reservoirs of fluoroquinolone-resistant (FQ-R) Escherichia coli of sequence type 131 (ST131). We screened for ST131 among LTCF residents, characterized isolates molecularly, and identified risk factors for colonization. Methods. We conducted a cross-sectional study using a single perianal swab or stool sample per resident in 2 LTCFs in Olmsted County, Minnesota, from April to July 2013. Confirmed FQ-R E. coli isolates underwent polymerase chain reaction-based phylotyping, detection of ST131 and its H30 and H30-Rx subclones, extended virulence genotyping, and pulsed-field gel electrophoresis (PFGE) analysis. Epidemiological data were collected from medical records. Results. Of 133 fecal samples, 33 (25%) yielded FQ-R E. coli, 32 (97%) of which were ST131. The overall proportion with ST131 intestinal colonization was 32 of 133 (24%), which differed by facility: 17 of 41 (42%) in facility 1 vs 15 of 92 (16%) in facility 2 (P = .002). All ST131 isolates represented the H30 subclone, with virulence gene and PFGE profiles resembling those of previously described ST131 clinical isolates. By PFGE, certain isolates clustered both within and across LTCFs. Multivariable predictors of ST131 colonization included inability to sign consent (odds ratio [OR], 4.16 [P = .005]), decubitus ulcer (OR, 4.87 [ P = .04]), and fecal incontinence (OR, 2.59 [P = .06]). Conclusions. Approximately one fourth of LTCF residents carried FQ-R ST131 E. coli resembling ST131 clinical isolates. Pulsed-field gel electrophoresis suggested intra- and interfacility transmission. The identified risk factors suggest that LTCF residents who require increased nursing care are at greatest risk for ST131 colonization, possibly due to healthcare-associated transmission.

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Outbreak of KPC-3-producing ST15 and ST348Klebsiella pneumoniaein a Portuguese hospital

Epidemiology and Infection ◽

10.1017/s0950268816002442 ◽

2016 ◽

Vol 145 (3) ◽

pp. 595-599 ◽

Cited By ~ 17

Author(s):

D. VUBIL ◽

R. FIGUEIREDO ◽

T. REIS ◽

C. CANHA ◽

L. BOAVENTURA ◽

...

Keyword(s):

Gel Electrophoresis ◽

Tertiary Care Hospital ◽

Tertiary Care ◽

Pulsed Field Gel Electrophoresis ◽

Care Hospital ◽

E Coli ◽

Carbapenemase Gene ◽

Sporadic Cases ◽

First Time ◽

Sequence Types

SUMMARYTo date, only a few sporadic cases of infections due toKlebsiella pneumoniaecarbapenemase (KPC) producers have been reported in Portugal. Here, we report for the first time an outbreak ofK. pneumoniaeKPC-3 producers in a tertiary-care hospital during 2013. Twenty-seven ertapenem-resistantK. pneumoniaewere identified in patients at a tertiary-care hospital during 2013 isolated predominantly from urine (48·1%) and blood (25·9%) cultures. All isolates were highly resistant toβ-lactam antibiotics and most showed intermediate resistance to imipenem. The more frequentβ-lactamases were TEM- (77·7%), CTX-M- (70·3%) and KPC-type (66·6%). KPC-3 was identified by sequencing. TheblaKPC−3gene was associated with an IncF plasmid, and efficiently transferred toE. coliJ53. Pulsed-field gel electrophoresis typing revealed three clusters of isolates which were further characterized by multi-locus sequence typing as ST11, ST15 and ST348. Ertapenem-resistant ST15 was already in circulation in the hospital, related to expression of OmpK36 modified porin, but the other two sequence types had not been previously found in the hospital. We conclude that the IncF plasmid mediated transfer of KPC-3 in the outbreak and that implementation of carbapenemase gene screening in isolates from patients on admission to hospital is advisable in order to control dissemination of these antimicrobial resistance elements.

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