scholarly journals The Pheno- and Genotypic Characterization of Porcine Escherichia coli Isolates

2021 ◽  
Vol 9 (8) ◽  
pp. 1676
Author(s):  
Tanja Bernreiter-Hofer ◽  
Lukas Schwarz ◽  
Elke Müller ◽  
Adriana Cabal-Rosel ◽  
Maciej Korus ◽  
...  

Escherichia (E.) coli is the main causative pathogen of neonatal and post-weaning diarrhea and edema disease in swine production. There is a significant health concern due to an increasing number of human infections associated with food and/or environmental-borne pathogenic and multidrug-resistant E. coli worldwide. Monitoring the presence of pathogenic and antimicrobial-resistant E. coli isolates is essential for sustainable disease management in livestock and human medicine. A total of 102 E. coli isolates of diseased pigs were characterized by antimicrobial and biocide susceptibility testing. Antimicrobial resistance genes, including mobile colistin resistance genes, were analyzed by PCR and DNA sequencing. The quinolone resistance-determining regions of gyrA and parC in ciprofloxacin-resistant isolates were analyzed. Clonal relatedness was investigated by two-locus sequence typing (CH clonotyping). Phylotyping was performed by the Clermont multiplex PCR method. Virulence determinants were analyzed by customized DNA-based microarray technology developed in this study for fast and economic molecular multiplex typing. Thirty-five isolates were selected for whole-genome sequence-based analysis. Most isolates were resistant to ampicillin and tetracycline. Twenty-one isolates displayed an ESBL phenotype and one isolate an AmpC β-lactamase-producing phenotype. Three isolates had elevated colistin minimal inhibitory concentrations and carried the mcr-1 gene. Thirty-seven isolates displayed a multi-drug resistance phenotype. The most predominant β-lactamase gene classes were blaTEM-1 (56%) and blaCTX-M-1 (13.71%). Mutations in QRDR were observed in 14 ciprofloxacin-resistant isolates. CH clonotyping divided all isolates into 51 CH clonotypes. The majority of isolates belonged to phylogroup A. Sixty-four isolates could be assigned to defined pathotypes wherefrom UPEC was predominant. WGS revealed that the most predominant sequence type was ST100, followed by ST10. ST131 was detected twice in our analysis. This study highlights the importance of monitoring antimicrobial resistance and virulence properties of porcine E. coli isolates. This can be achieved by applying reliable, fast, economic and easy to perform technologies such as DNA-based microarray typing. The presence of high-risk pathogenic multi-drug resistant zoonotic clones, as well as those that are resistant to critically important antibiotics for humans, can pose a risk to public health. Improved protocols may be developed in swine farms for preventing infections, as well as the maintenance and distribution of the causative isolates.

2021 ◽  
Vol 1 (1) ◽  
pp. 17-20
Author(s):  
Ahmed Abd El-Mawgoud ◽  
Azza El-Sawah ◽  
Soad Nasef ◽  
Al-Hussien Dahshan ◽  
Ahmed Ali

In the current study, ten avian pathogenic E. coli (APEC) isolates of the most predominant APEC serogroups isolated from broiler chickens in Egypt were screened for their virulence and antimicrobial resistance genes pattern using PCR. Five selected virulence gene patterns were further investigated for their in-vivo pathogenicity test. Results showed a 100% prevalence of the β-lactams and tetracyclines resistance genes. However, aminoglycoside and quinolone resistance genes were not detected. Also, 80% of the tested isolates harbored mcr-1 gene, colistin resistance gene. In-vivo pathogenic strains consistently harbored the virulence gene pattern of fimH, fimA, papC, iutA, and tsh. Additionally, the tsh gene was consistently detected with lethal APEC isolates in day-old chicks. These results highlighted the high prevalence of antimicrobial and virulence genes in APEC that potentially represent a public health concern. In this study, the virulence genes fimH, fimA, papC, iutA, and tsh were the most common virulence gene patterns associated with pathogenicity in day-old chicks.


2021 ◽  
Vol 12 ◽  
Author(s):  
Mohammed Elbediwi ◽  
Yanting Tang ◽  
Dawei Shi ◽  
Hazem Ramadan ◽  
Yaohui Xu ◽  
...  

Salmonella spp. is recognized as an important zoonotic pathogen. The emergence of antimicrobial resistance in Salmonella enterica poses a great public health concern worldwide. While the knowledge on the incidence and the characterization of different S. enterica serovars causing chick embryo death remains obscure in China. In this study, we obtained 45 S. enterica isolates from 2,139 dead chick embryo samples collected from 28 breeding chicken hatcheries in Henan province. The antimicrobial susceptibility assay was performed by the broth microdilution method and the results showed that 31/45 (68.8%) isolates were multidrug-resistant (≥3 antimicrobial classes). Besides the highest resistance rate was observed in the aminoglycoside class, all the isolates were susceptible to chloramphenicol, azithromycin, and imipenem. Furthermore, genomic characterization revealed that S. Enteritidis (33.33%; 15/45) was a frequent serovar that harbored a higher number of virulence factors compared to other serovars. Importantly, genes encoding β-lactamases were identified in three serovars (Thompson, Enteritidis, and Kottbus), whereas plasmid-mediated quinolone resistance genes (qnrB4) were detected in certain isolates of S. Thompson and the two S. Kottbus isolates. All the examined isolates harbored the typical virulence factors from Salmonella pathogenicity islands 1 and 2 (SPI-1 and SPI-2). Additionally, a correlation analysis between the antimicrobial resistance genes, phenotype, and plasmids was conducted among Salmonella isolates. It showed strong positive correlations (r < 0.6) between the different antimicrobial-resistant genes belonging to certain antimicrobial classes. Besides, IncF plasmid showed a strong negative correlation (r > −0.6) with IncHI2 and IncHI2A plasmids. Together, our study demonstrated antimicrobial-resistant S. enterica circulating in breeding chicken hatcheries in Henan province, highlighting the advanced approach, by using genomic characterization and statistical analysis, in conducting the routine monitoring of the emerging antimicrobial-resistant pathogens. Our findings also proposed that the day-old breeder chicks trading could be one of the potential pathways for the dissemination of multidrug-resistant S. enterica serovars.


2020 ◽  
Author(s):  
Saba Asgharzadeh Marghmalek ◽  
Reza Valadan ◽  
Mehrdad Gholami ◽  
Mohtaram Nasrolahei ◽  
Hamid Reza Goli

Abstract Background: The role of the hospital environment as a source of pathogenic bacteria in recent studies has been poorly investigated. This study investigated the distribution of antimicrobial resistance genes and virulence determinants in Enterococcus species isolated from hospital environment in Sari, Iran. A total of 90 enterococci isolates were identified and species identification confirmed with specific primers. Seven resistance genes and two virulence associated genes were evaluated molecularly by multiplex polymerase chain reaction. Results: Of the 90 enterococcal isolates, 42 (46.66%), and 48 (53.33%) were identified as E. faecalis, and E. faecium, respectively. Also, 28 (66.6%) E. faecalis and 18 (37.5%) E. faecium isolates were multidrug-resistant (MDR). Among all 90 environmental isolates 54 (60%), 54 (60%), 8 (8.8%), 8 (8.8%), 60 (66.6%), 26 (28.8%), and 24 (26.6%) isolates contained tetM, tetL, vanA, vanB, ermB, aac (6´)-Ie-aph (2´´)-Ia, and aph (3´)-IIIa, respectively. Moreover, 88 (97.7%) and 16 (17.7%) isolates were detected as esp and ace positive ones, correspondingly. Conclusions: This report showed that the environmental isolates of Enterococcus are the major sources of antibiotic resistance genes that can transfer them to the clinical isolates of bacteria in hospital settings. An effective following strategy should be organized to clearance and stop emergence of these pathogenic bacteria.


2019 ◽  
Author(s):  
Jingzhi Yuan ◽  
Xiaoye Wang ◽  
Dali Shi ◽  
Qiang Ge ◽  
Xingxing Song ◽  
...  

Abstract Background The discovery of mcr-1-positive Escherichia coli (MCRPEC), a notable superbug, attracted great attention worldwide. Swine-origin multi-drug resistance MCRPEC is a potential threat to public health and safety. To date, few detailed studies regarding swine-origin MCRPEC in Guangxi, South China, have been reported. Results In this study, thirty-three MCRPEC harbored mcr-1 genes were identified from 142 E. coli strains isolated from swine droppings and entrails in Guangxi in 2018. All MCRPEC isolates were assigned to 8 unique STs, including ST10, ST224 and ST410, which overlapped with the human-origin MCRPEC. Additionally, a total of six plasmid replicon types (IncFI, IncHI1, IncY, IncN, IncI1 and IncX1) were found. Moreover, the drug susceptibility of the MCRPEC isolates was tested with 27 antimicrobial agents belonging to 17 antimicrobial categories that are usually used in hospitals. There were 19 extended spectrum beta lactamase (ESBL) E. coli and 12 carbapenem resistant E. coli among the 33 MCRPEC strains. Importantly, the MCRPEC showed a high rate of resistance against two broad-spectrum carbapenem antibiotics, imipenem and meropenem, which are forbidden in livestock production use. Three MCRPEC strains were further identified to be extensively drug-resistant (XDR), and other isolates were recognized as multi-drug-resistant (MDR). Meanwhile, to detect whether plasmid-carrying antimicrobial resistance genes coexisted with the mcr-1 gene in the MCRPEC isolates, a total of 22 plasmid-carrying antimicrobial resistance genes were tested for. The results showed that four ESBL genes and one pAmpC gene were identified. Eight of the MCRPEC isolates also contained the carbapenem gene blaNDM-5, which could cause untreatable infections. Moreover, ten non-lactamase genes were also detected. Conclusion This study indicated that swine-origin MCRPEC isolated in Guangxi seemed to have a high rate of resistance to both regular and final line of defense drugs as well as drug resistance genes, which pose a great threat to human public safety and health.


Antibiotics ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1374
Author(s):  
Naiyaphat Nittayasut ◽  
Jitrapa Yindee ◽  
Pongthai Boonkham ◽  
Teerapong Yata ◽  
Nipattra Suanpairintr ◽  
...  

Resistance to extended-spectrum cephalosporins (ESC) and carbapenems in Escherichia coli (E. coli), increasingly identified in small animals, indicates a crisis of an antimicrobial resistance situation in veterinary medicine and public health. This study aimed to characterise the genetic features of ESC-resistant E. coli isolated from cats and dogs with urinary tract infections in Thailand. Of 72 ESC-resistant E. coli isolated from diagnostic samples (2016–2018), blaCTX-M including group 1 (CTX-M-55, -15 and -173) and group 9 (CTX-M-14, -27, -65 and -90) variants were detected in 47 isolates (65.28%) using PCR and DNA sequencing. Additional antimicrobial resistance genes, including plasmid-mediated AmpC (CIT and DHA), blaNDM-5, mcr-3, mph(A) and aac(6′)-Ib-cr, were detected in these isolates. Using a broth microdilution assay, all the strains exhibited multidrug-resistant phenotypes. The phylogroups were F (36.11%), A (20.83%), B1 (19.44%), B2 (19.44%) and D (4.17%), with several virulence genes, plasmid replicons and an integrase gene. The DNA fingerprinting using a repetitive extragenic palindromic sequence-PCR presented clonal relationships within phylogroups. Multiple human-associated, high-risk ExPEC clones associated with multidrug resistance, including sequence type (ST) 38, ST131, ST224, ST167, ST354, ST410, ST617 and ST648, were identified, suggesting clonal dissemination. Dogs and cats are a potential reservoir of ESC-resistant E. coli and significant antimicrobial resistance genes.


mSphere ◽  
2019 ◽  
Vol 4 (3) ◽  
Author(s):  
Liseth Salinas ◽  
Paúl Cárdenas ◽  
Timothy J. Johnson ◽  
Karla Vasco ◽  
Jay Graham ◽  
...  

ABSTRACT The increased prevalence of antimicrobial resistance (AMR) among Enterobacteriaceae has had major clinical and economic impacts on human medicine. Many of the multidrug-resistant (multiresistant) Enterobacteriaceae found in humans are community acquired, and some of them are possibly linked to food animals (i.e., livestock raised for meat and dairy products). In this study, we examined whether numerically dominant commensal Escherichia coli strains from humans (n = 63 isolates) and domestic animals (n = 174 isolates) in the same community and with matching phenotypic AMR patterns were clonally related or shared the same plasmids. We identified 25 multiresistant isolates (i.e., isolates resistant to more than one antimicrobial) that shared identical phenotypic resistance patterns. We then investigated the diversity of E. coli clones, AMR genes, and plasmids carrying the AMR genes using conjugation, replicon typing, and whole-genome sequencing. All of the multiresistant E. coli isolates (from children and domestic animals) analyzed had at least 90 or more whole-genome SNP differences between one another, suggesting that none of the strains was recently transferred. While the majority of isolates shared the same antimicrobial resistance genes and replicons, DNA sequencing indicated that these genes and replicons were found on different plasmid structures. We did not find evidence of the clonal spread of AMR in this community: instead, AMR genes were carried on diverse clones and plasmids. This presents a significant challenge for understanding the movement of AMR in a community. IMPORTANCE Even though Escherichia coli strains may share nearly identical phenotypic AMR profiles and AMR genes and overlap in space and time, the diversity of clones and plasmids challenges research that aims to identify sources of AMR. Horizontal gene transfer appears to play a more significant role than clonal expansion in the spread of AMR in this community.


Author(s):  
Katarzyna Ćwiek ◽  
Anna Woźniak-Biel ◽  
Magdalena Karwańska ◽  
Magdalena Siedlecka ◽  
Christine Lammens ◽  
...  

Abstract Background A plasmid-mediated mechanism of bacterial resistance to polymyxin is a serious threat to public health worldwide. The present study aimed to determine the occurrence of plasmid-mediated colistin resistance genes and to conduct the molecular characterization of mcr-positive Escherichia coli strains isolated from Polish poultry. Methods In this study, 318 E. coli strains were characterized by the prevalence of mcr1–mcr5 genes, antimicrobial susceptibility testing by minimal inhibitory concentration method, the presence of antimicrobial resistance genes was screened by PCR, and the biofilm formation ability was tested using the crystal violet staining method. Genetic relatedness of mcr-1-positive E. coli strains was evaluated by multilocus sequence typing method. Results Among the 318 E. coli isolates, 17 (5.35%) harbored the mcr-1 gene. High antimicrobial resistance rates were observed for ampicillin (100%), tetracycline (88.24%), and chloramphenicol (82.35%). All mcr-1-positive E. coli strains were multidrug-resistant, and as many as 88.24% of the isolates contained the blaTEM gene, tetracycline (tetA and tetB), and sulfonamide (sul1, sul2, and sul3) resistance genes. Additionally, 41.18% of multidrug-resistant, mcr-1-positive E. coli isolates were moderate biofilm producers, while the rest of the strains showed weak biofilm production. Nine different sequence types were identified, and the dominant ST was ST93 (29.41%), followed by ST117 (17.65%), ST156 (11.76%), ST 8979 (11.76%), ST744 (5.88%), and ST10 (5.88%). Moreover, the new ST was identified in this study. Conclusions Our results showed a low occurrence of mcr-1-positive E. coli strains isolated from Polish poultry; however, all the isolated strains were resistant to multiple antimicrobial agents and were able to form biofilms at low or medium level.


2017 ◽  
Author(s):  
Cameron J. Reid ◽  
Ethan R. Wyrsch ◽  
Piklu Roy Chowdhury ◽  
Tiziana Zingali ◽  
Michael Liu ◽  
...  

AbstractPorcine faecal waste is a serious environmental pollutant. Carriage of antimicrobial resistance and virulence-associated genes (VAGs) and the zoonotic potential of commensalEscherichia colifrom swine is largely unknown. Furthermore, little is known about the role of commensalE. colias contributors to the mobilisation of antimicrobial resistance genes between food animals and the environment. Here, we report whole genome sequence analysis of 141E. colifrom the faeces of healthy pigs. Most strains belonged to phylogroups A and B1 and carried i) a class 1 integron; ii) VAGs linked with extraintestinal infection in humans; iii) antimicrobial resistance genesblaTEM, aphAl, cmlA, strAB, tet(A)A,dfrA12, dfrA5, sul1, sul2, sul3; iv)IS26;and v) heavy metal resistance genes (merA, cusA, terA). Carriage of the sulphonamide resistance genesul3was notable in this study. The 141 strains belonged to 42 multilocus sequence types, but clonal complex 10 featured prominently. Structurally diverse class 1 integrons that were frequently associated with IS26 carried unique genetic features that were also identified in extraintestinal pathogenicE. coli(ExPEC) from humans. This study provides the first detailed genomic analysis and point of reference for commensalE. coliof porcine origin, facilitating tracking of specific lineages and the mobile resistance genes they carry.Conflict of Interest StatementNone to declare.


2019 ◽  
Author(s):  
Ana Carolina de Mello Santos ◽  
Rosa Maria Silva ◽  
Tiago Barcelos Valiatti ◽  
Fernanda Fernandes dos Santos ◽  
José Francisco Santos-Neto ◽  
...  

AbstractEscherichia coli EC121 is a multidrug-resistant (MDR) strain isolated from bloodstream infection of an inpatient with persistent gastroenteritis and Zone T lymphoma, that died due to septic shock. Despite causing an extraintestinal infection, it harbors few known virulence factors and was assigned into phylogenetic group B1. To evaluate if the EC121 was pathogenic or opportunistic, its genome was sequenced, and an in vitro characterization of some pathogenicity-associated properties was performed. The data retrieved from genome analysis showed that E. coli strain EC121 belongs to the O154:H25 serotype, and to ST101-B1, which was epidemiologically linked to extraintestinal infections and antimicrobial resistance spread as well. Moreover, it was closely related to Shiga-toxin producing E. coli (STEC). Besides, strain EC121 is an MDR strain harboring 14 antimicrobial resistance genes, including blaCTX-M-2, and more than 50 complete virulence genetic clusters, which are reported to be associated either with DEC or ExPEC. The strain also displays the capacity to adhere to a variety of cell lineages, and invade T24 bladder cells, as well as the ability to form biofilms on abiotic surfaces, and survive the bactericidal serum complement activity. Additionally, it is virulent in the Galleria mellonella model. Altogether, E. coli EC121 unveiled to be a pathogen powered by its multi-drug resistance characteristic. Carry out studies providing accurate information about the virulence potential of all kinds of MDR strains are essential because these studies will help in the development of alternative therapies of infection management and spread control of MDR strains.Authors summaryThe phylogenetic origin of extraintestinal pathogenic Escherichia coli is mostly associated with phylogroup B2, and the majority of the studies regarding extraintestinal infection focus on the most virulent strains, which might also present multidrug-resistant (MDR) phenotype. Strains belonging to phylogroup B1 and isolated from extraintestinal infections are considered as opportunist pathogens and have their virulence neglected. We focus our study in one MDR strain isolated from bloodstream infection that belongs to phylogenetic group B1 to enlarge the knowledge about the virulence of this kind of strain. We demonstrated that the EC121 is capable of adheres to intestinal and bladder human cells, and invades the latter one; it survives to human serum bactericidal effects and produces biofilm. Additionally, the in vivo assay confirmed the EC121 virulence, showing that it should be considered a pathogenic strain. The genetic analyzes highlighted important aspects of EC121 which are typical from strains of sequence type 101, like its involvement in the spread of antimicrobial resistance genes and its relationship with extraintestinal infection from diverse sources. Information concerning the virulence of MDR strains is important for the development of global actions treating the spread of antimicrobial resistance, as well as to elucidate the pathogenesis of strains that were considered as an opportunist.


2021 ◽  
Vol 9 (9) ◽  
pp. 1880
Author(s):  
Radwa Abdelwahab ◽  
Munirah M. Alhammadi ◽  
Ehsan A. Hassan ◽  
Entsar H. Ahmed ◽  
Nagla H. Abu-Faddan ◽  
...  

Klebsiella pneumoniae is an important human pathogen in both developing and industrialised countries that can causes a variety of human infections, such as pneumonia, urinary tract infections and bacteremia. Like many Gram-negative bacteria, it is becoming resistant to many frontline antibiotics, such as carbapenem and cephalosporin antibiotics. In Egypt, K. pneumoniae is increasingly recognised as an emerging pathogen, with high levels of antibiotic resistance. However, few Egyptian K. pneumoniae strains have been sequenced and characterised. Hence, here, we present the genome sequence of a multidrug resistant K. pneumoniae strain, KPE16, which was isolated from a child in Assiut, Egypt. We report that it carries multiple antimicrobial resistance genes, including a blaNDM-1 carbapenemase and extended spectrum β-lactamase genes (i.e., blaSHV-40, blaTEM-1B, blaOXA-9 and blaCTX-M-15). By comparing this strain with other Egyptian isolates, we identified common plasmids, resistance genes and virulence determinants. Our analysis suggests that some of the resistance plasmids that we have identified are circulating in K. pneumoniae strains in Egypt, and are likely a source of antibiotic resistance throughout the world.


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