scholarly journals Genomic Comparison of Conjugative Plasmids from Salmonella enterica and Escherichia coli Encoding Beta-Lactamases and Capable of Mobilizing Kanamycin Resistance Col-like Plasmids

2021 ◽  
Vol 9 (11) ◽  
pp. 2205
Author(s):  
Elizabeth A. McMillan ◽  
Ly-Huong T. Nguyen ◽  
Lari M. Hiott ◽  
Poonam Sharma ◽  
Charlene R. Jackson ◽  
...  

Salmonella enterica and Escherichia coli are important human pathogens that frequently contain plasmids, both large and small, carrying antibiotic resistance genes. Large conjugative plasmids are known to mobilize small Col plasmids, but less is known about the specificity of mobilization. In the current study, six S. enterica and four E. coli strains containing large plasmids were tested for their ability to mobilize three different kanamycin resistance Col plasmids (KanR plasmids). Large conjugative plasmids from five isolates, four S. enterica and one E. coli, were able to mobilize KanR plasmids of various types. Plasmids capable of mobilizing the KanR plasmids were either IncI1 or IncX, while IncI1 and IncX plasmids with no evidence of conjugation had disrupted transfer regions. Conjugative plasmids of similar types mobilized similar KanR plasmids, but not all conjugative plasmid types were capable of mobilizing all of the KanR plasmids. These data describe some of the complexities and specificities of individual small plasmid mobilization.

2003 ◽  
Vol 69 (8) ◽  
pp. 4915-4926 ◽  
Author(s):  
Michael B. Cooley ◽  
William G. Miller ◽  
Robert E. Mandrell

ABSTRACT Enteric pathogens, such as Salmonella enterica and Escherichia coli O157:H7, have been shown to contaminate fresh produce. Under appropriate conditions, these bacteria will grow on and invade the plant tissue. We have developed Arabidopsis thaliana (thale cress) as a model system with the intention of studying plant responses to human pathogens. Under sterile conditions and at 100% humidity, S. enterica serovar Newport and E. coli O157:H7 grew to 109 CFU g−1 on A. thaliana roots and to 2 × 107 CFU g−1 on shoots. Furthermore, root inoculation led to contamination of the entire plant, indicating that the pathogens are capable of moving on or within the plant in the absence of competition. Inoculation with green fluorescent protein-labeled S. enterica and E. coli O157:H7 showed invasion of the roots at lateral root junctions. Movement was eliminated and invasion decreased when nonmotile mutants of S. enterica were used. Survival of S. enterica serovar Newport and E. coli O157:H7 on soil-grown plants declined as the plants matured, but both pathogens were detectable for at least 21 days. Survival of the pathogen was reduced in unautoclaved soil and amended soil, suggesting competition from indigenous epiphytes from the soil. Enterobacter asburiae was isolated from soil-grown A. thaliana and shown to be effective at suppressing epiphytic growth of both pathogens under gnotobiotic conditions. Seed and chaff harvested from contaminated plants were occasionally contaminated. The rate of recovery of S. enterica and E. coli O157:H7 from seed varied from undetectable to 19% of the seed pools tested, depending on the method of inoculation. Seed contamination by these pathogens was undetectable in the presence of the competitor, Enterobacter asburiae. Sampling of 74 pools of chaff indicated a strong correlation between contamination of the chaff and seed (P = 0.025). This suggested that contamination of the seed occurred directly from contaminated chaff or by invasion of the flower or silique. However, contaminated seeds were not sanitized by extensive washing and chlorine treatment, indicating that some of the bacteria reside in a protected niche on the seed surface or under the seed coat.


Author(s):  
Mahbubul Siddiqee ◽  
Rebekah Henry ◽  
Rebecca Coulthard ◽  
Christelle Schang ◽  
Richard Williamson ◽  
...  

Estuarine bank sediments have the potential to support the survival and growth of fecal indicator organisms, including Escherichia coli. However, survival of fecal pathogens in estuarine sediments is not well researched and therefore remains a significant knowledge gap regarding public health risks in estuaries. In this study, simultaneous survival of Escherichia coli and a fecal pathogen, Salmonella enterica serovar Typhimurium, was studied for 21 days in estuarine bank sediment microcosms. Observed growth patterns for both organisms were comparable under four simulated scenarios; for continuous-desiccation, extended-desiccation, periodic-inundation, and continuous-inundation systems, logarithmic decay coefficients were 1.54/day, 1.51/day, 0.14/day, and 0.20/day, respectively, for E. coli, and 1.72/day, 1.64/day, 0.21/day, and 0.24/day for S. Typhimurium. Re-wetting of continuous-desiccated systems resulted in potential re-growth, suggesting survival under moisture-limited conditions. Key findings from this study include: (i) Bank sediments can potentially support human pathogens (S. Typhimurium), (ii) inundation levels influence the survival of fecal bacteria in estuarine bank sediments, and (iii) comparable survival rates of S. Typhimurium and E. coli implies the latter could be a reliable fecal indicator in urban estuaries. The results from this study will help select suitable monitoring and management strategies for safer recreational activities in urban estuaries.


2013 ◽  
Vol 79 (15) ◽  
pp. 4613-4619 ◽  
Author(s):  
Patrick Studer ◽  
Werner E. Heller ◽  
Jörg Hummerjohann ◽  
David Drissner

ABSTRACTSprouts contaminated with human pathogens are able to cause food-borne diseases due to the favorable growth conditions for bacteria during germination and because of minimal processing steps prior to consumption. We have investigated the potential of hot humid air, i.e., aerated steam, to treat alfalfa and mung bean seeds which have been artificially contaminated withEscherichia coliO157:H7,Salmonella entericasubsp.entericaserovar Weltevreden, andListeria monocytogenesScott A. In addition, a recently collectedE. coliO178:H12 isolate, characterized by a reduced heat sensitivity, was exposed to the treatment described. Populations ofE. coliO157:H7 andS. entericaon alfalfa and mung bean seeds could be completely eliminated by a 300-s treatment with steam at 70 ± 1°C as revealed by enrichment studies.L. monocytogenesandE. coliO178:H12 could not be completely eliminated from artificially inoculated seeds. However, bacterial populations were reduced by more than 5 log CFU/g on alfalfa and by more than 4 log CFU/g on mung bean seeds. The germination rate of mung beans was not affected by the 300-s treatment compared to the germination rate of untreated seeds whereas that of alfalfa seeds was significantly lower by 11.9%. This chemical-free method is an effective alternative to the 20,000-ppm hypochlorite treatment presently recommended by the U.S. Food and Drug Administration (FDA).


2002 ◽  
Vol 68 (6) ◽  
pp. 3114-3120 ◽  
Author(s):  
A. O. Charkowski ◽  
J. D. Barak ◽  
C. Z. Sarreal ◽  
R. E. Mandrell

ABSTRACT Sprout producers have recently been faced with several Salmonella enterica and Escherichia coli O157:H7 outbreaks. Many of the outbreaks have been traced to sprout seeds contaminated with low levels of human pathogens. Alfalfa seeds were inoculated with S. enterica and E. coli O157:H7 strains isolated from alfalfa seeds or other environmental sources and sprouted to examine growth of these human pathogens in association with sprouting seeds. S. enterica strains grew an average of 3.7 log10 on sprouting seeds over 2 days, while E. coli O157:H7 strains grew significantly less, an average of 2.3 log10. The initial S. enterica or E. coli O157:H7 inoculum dose and seed-sprouting temperature significantly affected the levels of both S. enterica and E. coli O157:H7 on the sprouts and in the irrigation water, while the frequency of irrigation water replacement affected only the levels of E. coli O157:H7. Colonization of sprouting alfalfa seeds by S. enterica serovar Newport and E. coli O157:H7 strains transformed with a plasmid encoding the green fluorescent protein was examined with fluorescence microscopy. Salmonella serovar Newport colonized both seed coats and sprout roots as aggregates, while E. coli O157:H7 colonized only sprout roots.


2005 ◽  
Vol 71 (10) ◽  
pp. 6165-6174 ◽  
Author(s):  
Eelco Franz ◽  
Anne D. van Diepeningen ◽  
Oscar J. de Vos ◽  
Ariena H. C. van Bruggen

ABSTRACT Survival of the green fluorescent protein-transformed human pathogens Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium was studied in a laboratory-simulated lettuce production chain. Dairy cows were fed three different roughage types: high-digestible grass silage plus maize silage (6:4), low-digestible grass silage, and straw. Each was adjusted with supplemental concentrates to high and low crude protein levels. The pathogens were added to manure, which was subsequently mixed (after 56 and 28 days for E. coli O157:H7 and Salmonella serovar Typhimurium, respectively) with two pairs of organically and conventionally managed loamy and sandy soil. After another 14 days, iceberg lettuce seedlings were planted and then checked for pathogens after 21 days of growth. Survival data were fitted to a logistic decline function (exponential for E. coli O157:H7 in soil). Roughage type significantly influenced the rate of decline of E. coli O157:H7 in manure, with the fastest decline in manure from the pure straw diet and the slowest in manure from the diet of grass silage plus maize silage. Roughage type showed no effect on the rate of decline of Salmonella serovar Typhimurium, although decline was significantly faster in the manure derived from straw than in the manure from the diet of grass silage plus maize silage. The pH and fiber content of the manure were significant explanatory factors and were positively correlated with the rate of decline. With E. coli O157:H7 there was a trend of faster decline in organic than in conventional soils. No pathogens were detected in the edible lettuce parts. The results indicate that cattle diet and soil management are important factors with respect to the survival of human pathogens in the environment.


2009 ◽  
Vol 54 (2) ◽  
pp. 590-596 ◽  
Author(s):  
Douglas R. Call ◽  
Randall S. Singer ◽  
Da Meng ◽  
Shira L. Broschat ◽  
Lisa H. Orfe ◽  
...  

ABSTRACT Large multidrug resistance plasmids of the A/C incompatibility complex (IncA/C) have been found in a diverse group of Gram-negative commensal and pathogenic bacteria. We present three completed sequences from IncA/C plasmids that originated from Escherichia coli (cattle) and Salmonella enterica serovar Newport (human) and that carry the cephamycinase gene bla CMY-2. These large plasmids (148 to 166 kbp) share extensive sequence identity and synteny. The most divergent plasmid, peH4H, has lost several conjugation-related genes and has gained a kanamycin resistance region. Two of the plasmids (pAM04528 and peH4H) harbor two copies of bla CMY-2, while the third plasmid (pAR060302) harbors a single copy of the gene. The majority of single-nucleotide polymorphisms comprise nonsynonymous mutations in floR. A comparative analysis of these plasmids with five other published IncA/C plasmids showed that the bla CMY-2 plasmids from E. coli and S. enterica are genetically distinct from those originating from Yersinia pestis and Photobacterium damselae and distal to one originating from Yersinia ruckeri. While the overall similarity of these plasmids supports the likelihood of recent movements among E. coli and S. enterica hosts, their greater divergence from Y. pestis or Y. ruckeri suggests less recent plasmid transfer among these pathogen groups.


2009 ◽  
Vol 72 (1) ◽  
pp. 21-27 ◽  
Author(s):  
SUPAKANA NAGACHINTA ◽  
JINRU CHEN

This study was undertaken to characterize the integrons present in a group of Shiga toxin–producing Escherichia coli (STEC) isolates and the ability of these integrons to transfer antibiotic resistance genes from STEC to E. coli K-12 MG1655. A total of 177 STEC isolates were analyzed for antibiotic susceptibility and the presence of integrons. Class 1 integrons were detected in 14 STEC isolates, and a class 2 integron was identified in 1 STEC isolate. The STEC isolates positive for class 1 integrons were resistant to streptomycin (MICs > 128 μg/ml) and sulfisoxazole (MICs > 1,024 μg/ml), and the isolate positive for the class 2 integron was resistant to streptomycin (MIC of 128 μg/ml), trimethoprim (MIC > 256 μg/ml), and streptothricin (MIC > 32 μg/ml). Results of restriction digestion and nucleotide sequencing revealed that the cassette regions of the class 1 integrons had a uniform size of 1.1 kb and contained a nucleotide sequence identical to that of aadA1. The class 2 integron cassette region was 2.0 kb and carried nucleotide sequences homologous to those of aadA1, sat1, and dfrA1. Results of the conjugation experiments revealed that horizontal transfers of conjugative plasmids are responsible for the dissemination of class 1 integron–mediated antibiotic resistance genes from STEC to E. coli K-12 MG1655. Antibiotic resistance traits not mediated by integrons, such as resistance to tetracycline and oxytetracycline, were cotransferred with the integron-mediated antibiotic resistance genes. The study suggested a possible role of integron and conjugative plasmid in dissemination of genes conferring resistance to antibiotics from pathogenic to generic E. coli cells.


2021 ◽  
Vol 4 (1) ◽  
pp. 25-29
Author(s):  
Diwakar Kumar ◽  
Ananya Choudhury ◽  
Mitul Nath ◽  
Udaya Kumar Vandana

At present, extended-spectrum β-lactamase (ESBL)-producing Escherichia coli and other species of Enterobacteriaceae have emerged as a matter of utmost concern. Indiscriminate utilization of antimicrobials in aquaculture generates selective pressure creating reservoirs of drug-resistance genes in fish pathogens and other bacteria, which may disseminate by horizontal gene transfer and reach human pathogens. The present study aims to detect extended-spectrum beta-lactamases (ESBL) production by enterobacteria isolated from locally harbored and imported fresh and dry fishes. Out of 235 fish samples investigated, the observed incidence of 9.78% (n=23) E. coli isolates. PCR detection of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli using primers specific for Bla-TEM, Bla-SHV, Bla-CTX-M. As per molecular detection by Multiplex PCR, 11 isolates exhibited Bla-TEM+CTX-M+SHV genes, and two isolates exhibited the presence of TEM genes only. Antimicrobial susceptibility patterns against 12 antibiotics were studied, where it was observed that resistance to the selected drug ranges from 4.34 % to 73.9 % for 12 different antibiotics.


2005 ◽  
Vol 49 (1) ◽  
pp. 71-76 ◽  
Author(s):  
Hedi Mammeri ◽  
Marc Van De Loo ◽  
Laurent Poirel ◽  
Luis Martinez-Martinez ◽  
Patrice Nordmann

ABSTRACT Although quinolone resistance results mostly from chromosomal mutations, it may also be mediated by a plasmid-encoded qnr gene in members of the family Enterobacteriaceae. Thus, 297 nalidixic-acid resistant strains of 2,700 Escherichia coli strains that had been isolated at the Bicêtre Hospital (Le Kremlin-Bicêtre, France) in 2003 were screened for qnr by PCR. A single E. coli isolate that carried a ca. 180-kb conjugative plasmid encoding a qnr determinant was identified. It conferred low-level resistance to quinolones and was associated with a chromosomal mutation in subunit A of the topoisomerase II gene. The qnr gene was located on a sul1-type class 1 integron just downstream of a conserved region (CR) element (CR1) comprising the Orf513 recombinase. Promoter sequences for qnr expression overlapped the extremity of CR1, indicating the role of CR1 in the expression of antibiotic resistance genes. This integron was different from other qnr-positive sul1-type integrons identified in American and Chinese enterobacterial isolates. In addition, plasmid pQR1 carried another class 1 integron that was identical to In53 from E. coli. The latter integron possessed a series of gene cassettes, including those coding for the extended-spectrum β-lactamase VEB-1, the rifampin ADP ribosyltransferase ARR-2, and several aminoglycoside resistance markers. This is the first report of plasmid-mediated quinolone resistance in Europe associated with an unknown level of plasmid-mediated multidrug resistance in Enterobacteriaceae.


2016 ◽  
Vol 60 (4) ◽  
pp. 2450-2455 ◽  
Author(s):  
Miaomiao Xie ◽  
Dachuan Lin ◽  
Kaichao Chen ◽  
Edward Wai Chi Chan ◽  
Wen Yao ◽  
...  

ABSTRACTA total of 55 cefotaxime-resistantEscherichia coliisolates were obtained from retail meat products purchased in Shenzhen, China, during the period November 2012 to May 2013. Thirty-seven of these 55 isolates were found to harbor ablaCTX-Mgene, with theblaCTX-M-1group being the most common type.blaCMY-2was detected in 16 isolates, alone or in combination with other extended-spectrum β-lactamase (ESBL) determinants. Importantly, thefosA3gene, which encodes fosfomycin resistance, was detected in 12 isolates, with several being found to reside in the conjugative plasmid that harbored theblaCTX-Mgene. The insertion sequence IS26was observed upstream of some of theblaCTX-M-55andfosA3genes. Conjugation experiments showed thatblaCTX-Mgenes from 15 isolates were transferrable, with Inc I1 and Inc FII being the most prevalent replicons. High clonal diversity was observed among theblaCTX-Mproducers, suggesting that horizontal transfer of theblaCTX-Mgenes amongE. colistrains in retail meats is a common event and that such strains may constitute an important reservoir ofblaCTX-Mgenes, which may be readily disseminated to other potential human pathogens.


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