Studies of Virulence Factors of Avian Pathogenic Escherichia coli in Avian Colibacillosis by In vitro Method and PCR

2021 ◽  
Author(s):  
Ayushi Singh ◽  
Daljeet Chhabra ◽  
Rakhi Gangil ◽  
Rakesh Sharda ◽  
Ravi Sikrodia ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Molecular Detection ◽  
Bacterial Disease ◽  
Avian Pathogenic Escherichia Coli ◽  
E Coli ◽  
Pathological Conditions ◽  
Biofilm Production ◽  
Pathogenic Escherichia Coli ◽  
Stx1 Gene

Background: Avian colibacillosis is considered as major cause of morbidity and mortality in poultry. It is a common bacterial disease of poultry and many virulence factors of E. coli are associated with the disease. The current study was aimed to investigate the presence of some virulence factors of E. coli isolated from the cases of colibacillosis.Methods: In present study, total 150 samples (liver, heart, lungs, air sacs and feaces) of chicken exhibiting pathological conditions of colibacillosis were collected from various poultry farms (organized and backyard) situated in and around Mhow and Indore cities. E.coli was isolated and identified from the samples on the basis of cultural characteristics and biochemical test. All E. coli isolates were further subjected to evaluate the presence of virulence factors such as biofilm production, haemolysis, invasiveness and molecular detection of fimH and stx1 gene.Result: Out of these 51.33% of incidence of E. coli was recorded. E. coli O84 and O149 serotypes were found most prevalent. Out of 77 isolates, 46 (59.7%) and 45 (58.4%) were positive for biofilm formation by tube method and modified CRA method, respectively. All E. coli isolates were showing invasiveness in congo red binding assay while none of the isolates was found haemolytic. Molecular detection revealed the presence of fimH (508bp) gene in 33.3% of tested samples while stx1 gene could not be detected in any isolates.

scholarly journals Multidrug-Resistant Avian Pathogenic Escherichia coli Strains and Association of Their Virulence Genes in Bangladesh

2020 ◽  
Vol 8 (8) ◽  
pp. 1135
Author(s):  
Otun Saha ◽  
M. Nazmul Hoque ◽  
Ovinu Kibria Islam ◽  
Md. Mizanur Rahaman ◽  
Munawar Sultana ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Virulence Genes ◽  
Multidrug Resistant ◽  
Avian Pathogenic Escherichia Coli ◽  
E Coli ◽  
Molecular Approaches ◽  
Clade B ◽  
Pathogenic Escherichia Coli ◽  
Resistance Patterns

The avian pathogenic Escherichia coli (APEC) strains are the chief etiology of colibacillosis worldwide. The present study investigated the circulating phylotypes, existence of virulence genes (VGs), and antimicrobial resistance (AMR) in 392 APEC isolates, obtained from 130 samples belonged to six farms using both phenotypic and PCR-based molecular approaches. Congo red binding (CRB) assay confirmed 174 APEC isolates which were segregated into ten, nine, and eight distinct genotypes by RAPD assay (discriminatory index, DI = 0.8707), BOX-PCR (DI = 0.8591) and ERIC-PCR (DI = 0.8371), respectively. The combination of three phylogenetic markers (chuA, yjaA and DNA fragment TspE4.C2) classified APEC isolates into B23 (37.36%), A1 (33.91%), D2 (11.49%), B22 (9.20%), and B1 (8.05%) phylotypes. Majority of the APEC isolates (75–100%) harbored VGs (ial, fimH, crl, papC, and cjrC). These VGs (papC and cjrC) and phylotypes (D2 and B2) of APEC had significant (p = 0.004) association with colibacillosis. Phylogenetic analysis showed two distinct clades (clade A and clade B) of APEC, where clade A had 98–100% similarity with E. coli APEC O78 and E. coli EHEC strains, and clade B had closest relationship with E. coli O169:H41 strain. Interestingly, phylogroups B2 and D2 were found in the APEC strains of both clades, while the strains from phylogroups A1 and B1 were found in clade A only. In this study, 81.71% of the isolates were biofilm formers, and possessed plasmids of varying ranges (1.0 to 54 kb). In vitro antibiogram profiling revealed that 100% isolates were resistant to ≥3 antibiotics, of which 61.96%, 55.24%, 53.85%, 51.16% and 45.58% isolates in phylotypes B1, D2, B22, B23, and A1, respectively, were resistant to these antimicrobials. The resistance patterns varied among different phylotypes, notably in phylotype B22, showing the highest resistance to ampicillin (90.91%), nalidixic acid (90.11%), tetracycline (83.72%), and nitrofurantoin (65.12%). Correspondence analysis also showed significant correlation among phylotypes with CRB (p = 0.008), biofilm formation (p = 0.02), drug resistance (p = 0.03), and VGs (p = 0.06). This report demonstrated that B2 and A1 phylotypes are dominantly circulating APEC phylotypes in Bangladesh; however, B2 and D2 are strongly associated with the pathogenicity. A high prevalence of antibiotic-resistant APEC strains from different phylotypes suggest the use of organic antimicrobial compounds, and/or metals, and the rotational use of antibiotics in poultry farms in Bangladesh.

scholarly journals Sub-lethal Concentrations of Phytochemicals (Carvacrol and Oregano) Select for Reduced Susceptibility Mutants of Escherichia coli O23:H52

2020 ◽  
Vol 69 (1) ◽  
pp. 121-125
Author(s):  
AFNAN A. AL-MNASER ◽  
MARTIN J. WOODWARD
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Coli Strain ◽  
Whole Genome ◽  
Avian Pathogenic Escherichia Coli ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Pathogenic Escherichia Coli ◽  
Survival Mechanisms

In vitro studies aimed at studying the mechanism of action of carvacrol and oregano as natural anti-bacterial agents to control multiple antibiotic-resistant avian pathogenic Escherichia coli (APEC) strain O23:H52 isolated from chicken were performed. Derivatives with increased minimum inhibitory concentrations (MIC) to the phytochemicals were selected after growing Escherichia coli (E. coli) strain O23:H52 at sub-lethal concentrations of carvacrol and oregano for a period of 60 days. Whole-genome sequencing (WGS) of two derivatives revealed a missense mutation in cadC and marR: the genes responsible for survival mechanisms and antibiotic resistance by efflux, respectively.

scholarly journals CHARACTERIZATION OF ENTEROAGGREGATIVE ESCHERICHIA COLI AMONG DIARRHEAL CHILDRENIN WESTERN BRAZILIAN AMAZON

2018 ◽  
Vol 55 (4) ◽  
pp. 390-396 ◽  
Author(s):  
Roger Lafontaine Mesquita TABORDA ◽  
Luiz Antônio da SILVA ◽  
Patricia Puccinelli ORLANDI ◽  
Flávia Serrano BATISTA ◽  
Renata Santos RODRIGUES ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Virulence Factors ◽  
Antimicrobial Susceptibility ◽  
Antibiotic Sensitivity ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Biofilm Production ◽  
Porto Velho

ABSTRACT BACKGROUND: Enteroaggregative Escherichia coli (EAEC) is one of the main acute and chronic diarrhea causes both in children and adults, mainly in developing countries. OBJECTIVE: The aim of the present study is to characterize EAEC strains isolated from faecal samples and to identify genes potentially contributing to virulence, biofilm production and antimicrobial resistance in children admitted to a pediatric hospital in Porto Velho, Rondônia State. METHODS: The total of 1,625 E. coli specimens were isolated from 591 children in the age group 6 years or younger who were hospitalized in Cosme and Damião Children Hospital in Porto Velho, between February 2010 and February 2012, with acute gastroenteritis. Colonies suggestive of E. coli were subjected to polymerase chain reaction testing in order to identify the virulence factors. The in vitro adhesion assays using HEp-2 adherence were tests. Biofilm detection through spectrophotometry and antimicrobial susceptibility tests were conducted in the disk diffusion method. RESULTS: The mentioned study examined 591 stool samples from children with diarrhea. Diarrheogenic E. coli was found in 27.4% (162/591) of the children. EAEC was the diarreagenic E. coli most frequently associated with diarrhea 52.4% (85/162), which was followed by enteropathogenic E. coli 43.8% (71/162), enterotoxigenic E. coli 2.4% (4/162), and enterohemorrhagic E. coli 1.2% (2/162). The aggR gene was detected in 63.5% (54/85) of EAEC isolates; moreover, statistically significant correlation was observed among typical EAEC (aggR) and aatA (P<0.0001), irp2 (P=0.0357) and shf (P=0.0328). It was recorded that 69% (59/85) of the 85 analyzed EAEC strains were biofilm producers; 73% (43/59) of the biofilm producers carried the aggR gene versus 42.3% (11/26) of non-producers (P=0.0135). In addition, there was association between the aatA gene and biofilm production; 61% (36/59) of the samples presented producer strains, versus 19.2% (5/26) of non-producers (P<0.0004). Antibiotic sensitivity test evidenced that most EAEC were ampicillin 70.6% (60/85), sulfamethoxazole 60% (51/85), tetracycline 44.7% (38/85) and cefotaxime 22.4% (19/85) resistant. CONCLUSION: As far as it is known, the present study is pioneer in Northern Brazil to investigate EAEC virulence factors and to show the antimicrobial susceptibility of EAEC strains isolated from children with diarrhea.

Acetate Kinase (AcK) is Essential for Microbial Growth and Betel-derived Compounds Potentially Target AcK, PhoP and MDR Proteins in M. tuberculosis, V. cholerae and Pathogenic E. coli: An in silico and in vitro Study

2019 ◽  
Vol 18 (31) ◽  
pp. 2731-2740 ◽  
Author(s):  
Sandeep Tiwari ◽  
Debmalya Barh ◽  
M. Imchen ◽  
Eswar Rao ◽  
Ranjith K. Kumavath ◽  
...  
Keyword(s):  
Broad Spectrum ◽  
In Silico ◽  
Pathogenic Bacteria ◽  
In Vitro Study ◽  
Docking Studies ◽  
Acetate Kinase ◽  
E Coli ◽  
Pathogenic Escherichia Coli ◽  
Novel Target

Background: Mycobacterium tuberculosis, Vibrio cholerae, and pathogenic Escherichia coli are global concerns for public health. The emergence of multi-drug resistant (MDR) strains of these pathogens is creating additional challenges in controlling infections caused by these deadly bacteria. Recently, we reported that Acetate kinase (AcK) could be a broad-spectrum novel target in several bacteria including these pathogens. Methods: Here, using in silico and in vitro approaches we show that (i) AcK is an essential protein in pathogenic bacteria; (ii) natural compounds Chlorogenic acid and Pinoresinol from Piper betel and Piperidine derivative compound 6-oxopiperidine-3-carboxylic acid inhibit the growth of pathogenic E. coli and M. tuberculosis by targeting AcK with equal or higher efficacy than the currently used antibiotics; (iii) molecular modeling and docking studies show interactions between inhibitors and AcK that correlate with the experimental results; (iv) these compounds are highly effective even on MDR strains of these pathogens; (v) further, the compounds may also target bacterial two-component system proteins that help bacteria in expressing the genes related to drug resistance and virulence; and (vi) finally, all the tested compounds are predicted to have drug-like properties. Results and Conclusion: Suggesting that, these Piper betel derived compounds may be further tested for developing a novel class of broad-spectrum drugs against various common and MDR pathogens.

scholarly journals Avian Pathogenic Escherichia coli (APEC): An Overview of Virulence and Pathogenesis Factors, Zoonotic Potential, and Control Strategies

Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 467
Author(s):  
Dipak Kathayat ◽  
Dhanashree Lokesh ◽  
Sochina Ranjit ◽  
Gireesh Rajashekara
Keyword(s):  
Escherichia Coli ◽  
Current Status ◽  
Neonatal Meningitis ◽  
Clear Understanding ◽  
Zoonotic Potential ◽  
Secretion Systems ◽  
Avian Pathogenic Escherichia Coli ◽  
E Coli ◽  
Pathogenic Escherichia Coli ◽  
Tract Infections

Avian pathogenic Escherichia coli (APEC) causes colibacillosis in avian species, and recent reports have suggested APEC as a potential foodborne zoonotic pathogen. Herein, we discuss the virulence and pathogenesis factors of APEC, review the zoonotic potential, provide the current status of antibiotic resistance and progress in vaccine development, and summarize the alternative control measures being investigated. In addition to the known virulence factors, several other factors including quorum sensing system, secretion systems, two-component systems, transcriptional regulators, and genes associated with metabolism also contribute to APEC pathogenesis. The clear understanding of these factors will help in developing new effective treatments. The APEC isolates (particularly belonging to ST95 and ST131 or O1, O2, and O18) have genetic similarities and commonalities in virulence genes with human uropathogenic E. coli (UPEC) and neonatal meningitis E. coli (NMEC) and abilities to cause urinary tract infections and meningitis in humans. Therefore, the zoonotic potential of APEC cannot be undervalued. APEC resistance to almost all classes of antibiotics, including carbapenems, has been already reported. There is a need for an effective APEC vaccine that can provide protection against diverse APEC serotypes. Alternative therapies, especially the virulence inhibitors, can provide a novel solution with less likelihood of developing resistance.

Serological diversity, molecular characterisation and antimicrobial sensitivity of avian pathogenic Escherichia coli (APEC) isolates from broiler chickens in Kashmir, India

2019 ◽  
Vol 59 (2) ◽  
pp. 338
Author(s):  
S. N. Magray ◽  
S. A. Wani ◽  
Z. A. Kashoo ◽  
M. A. Bhat ◽  
S. Adil ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Broiler Chickens ◽  
Virulence Genes ◽  
Virulence Gene ◽  
Gene Profile ◽  
Avian Pathogenic Escherichia Coli ◽  
Pathogenic Escherichia Coli ◽  
Significant Difference ◽  
Virulence Gene Profile

The present study has determined the serological diversity, virulence-gene profile and in vitro antibiogram of avian pathogenic Escherichia coli (APEC) isolates from broiler chickens in India suspected to have died of colibacillosis. The virulence-gene profile of APEC was compared with that of the Escherichia coli isolates from faeces of apparently healthy chickens, called avian faecal E. coli (AFEC). In total, 90 representative isolates of APEC and 63 isolates of AFEC were investigated in the present study. The APEC were typed into 19 serogroups, while some isolates were rough and could not be typed. Most prevalent serogroup was O2 (24.44%). Among the eight virulence genes studied, the prevalence of seven genes (iss, iucD, tsh, cva/cvi, irp2, papC and vat) was significantly higher in APEC than in AFEC isolates. However, there was no significant difference between APEC and AFEC isolates for possession of astA gene. The most frequent gene detected among the two groups of organisms was iss, which was present in 98.88% and 44.44% of APEC and AFEC isolates respectively. The in vitro antibiogram showed that the majority (96.6%) of APEC isolates were resistant to tetracycline, while 82.2% were resistant to cephalexin, 78.8% to cotrimoxazole, 68.8% to streptomycin and 63.3% to ampicillin. However, most of them (84.45%) were sensitive to gentamicin. Thus, it is concluded that APEC from the broiler chickens carried putative virulence genes that attributed to their pathogenicity. Furthermore, the majority of APEC isolates were found to be multi-drug resistant, which, in addition to leading treatment failures in poultry, poses a public health threat.

scholarly journals Molecular Detection of Avian Pathogenic Escherichia coli (APEC) for the First Time in Layer Farms in Bangladesh and Their Antibiotic Resistance Patterns

2020 ◽  
Vol 8 (7) ◽  
pp. 1021 ◽  
Author(s):  
Samina Ievy ◽  
Md. Saiful Islam ◽  
Md. Abdus Sobur ◽  
Mithun Talukder ◽  
Md. Bahanur Rahman ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Virulence Genes ◽  
Molecular Detection ◽  
Avian Pathogenic Escherichia Coli ◽  
Internal Organs ◽  
Pathogenic Escherichia Coli ◽  
Resistance Patterns ◽  
Antibiotic Resistance Patterns ◽  
First Time

Avian pathogenic Escherichia coli (APEC) causes significant economic losses in poultry industries. Here, we determined for the first time in Bangladesh, the prevalence of APEC-associated virulence genes in E. coli isolated from layer farms and their antibiotic resistance patterns. A total of 99 samples comprising internal organs, feces, and air were collected from 32 layer farms. Isolation was performed by culturing samples on eosin–methylene blue agar plates, while the molecular detection of APEC was performed by PCR, and antibiograms were performed by disk diffusion. Among the samples, 36 were positive for the APEC-associated virulence genes fimC, iucD, and papC. Out of 36 isolates, 7, 18, and 11 were positive, respectively, for three virulence genes (papC, fimC, and iucD), two virulence genes, and a single virulence gene. Although the detection of virulence genes was significantly higher in the internal organs, the air and feces were also positive. The antibiograms revealed that all the isolates (100%) were resistant to ampicillin and tetracycline; 97.2%, to chloramphenicol and erythromycin; 55.5%, to enrofloxacin; 50.0%, to norfloxacin and ciprofloxacin; 19.4%, to streptomycin; 11.1%, to colistin; and 8.33%, to gentamicin. Interestingly, all the isolates were multidrug-resistant (MDR). Spearman’s rank correlation coefficient analysis revealed the strongest significant correlation between norfloxacin and ciprofloxacin resistance. This is the first study in Bangladesh describing the molecular detection of APEC in layer farms. Isolated APEC can now be used for detailed genetic characterization and assessing the impact on public health.

scholarly journals 1127. Genomic Analysis of Biofilm-Forming Enteroinvasive E. coli Emergent Pathogen

2018 ◽  
Vol 5 (suppl_1) ◽  
pp. S337-S338
Author(s):  
Oscar Gomez-Duarte ◽  
Julio Guerra ◽  
Ricky Ko
Keyword(s):  
Virulence Factors ◽  
Dna Sequences ◽  
Horizontal Transfer ◽  
Antibiotic Resistance Genes ◽  
Genomic Analysis ◽  
Coli Strain ◽  
Virulence Plasmid ◽  
E Coli ◽  
Query Coverage

Abstract Background Enteroinvasive Escherichia coli (EIEC) are involved in dysenteric diarrhea among children in low- and middle-income countries. EIEC strains isolated in Colombia, South America were shown to form biofilms and to be invasive in vitro. The O96:H19 serotypes and biofilm formation (BF) are not common phenotypes among EIEC, and the role they may play in diarrhea is at present unknown. The main goal of this study was to identify virulence and BF genes from EIEC genomic data. We hypothesize that EIEC O96:H19 strain 52.1 originated from horizontal transfer of a Shigella-like virulence plasmid into a non-EIEC pathogenic E coli strain. Methods WGS was performed on the BF-EIEC 52.1 strain using NextGen Illumina and Pacific Biosciences (PacBio) platforms. Publically available genomes from other EIEC O96H19 and Shigella genomes previously published were analyzed using online available software and databases including NCBI, BLAST, Mauve, among others. This analysis was tailored to identify virulence factors from the virulence factor database (VFDB). BLASTn was used to determine identity and query coverage of genes encoding the Shigella virulence factors. EIEC and Shigella genomes were analyzed on a multiple genome alignment software (Mauve) to verify results from BLASTn and to determine pseudogenes. Results The genome of EIEC O96:H19 strain 52.1 was 5,193,449 bp in size, containing 5,050 coding DNA sequences (CDSs). O96:H19 strain 52.1 carries three plasmids, the invasion plasmid (pINV) contains all type 3 secretion system (TTSS) and TTSS effectors genes previously described for Shigella and EIEC O96:H19 CFSAN029787 Italian strain. Non-TTSS virulence genes were also identified, including: long polar fimbrial gene (IpfA), enterotoxin (senB), and antibiotic resistance genes. Conclusion The EIEC O96:H19 strain 52.1 genome carries TTSS genes within a virulence plasmid, protein effector genes, and enterotoxin genes known to be associated with EIEC virulence. The EIEC O96:H19 stain 52.1 is an emergent diarrheagenic pathogen likely derived from an E. coli O96:H19 strain that acquired a Shigella-like virulence plasmid by horizontal transfer. Disclosures All authors: No reported disclosures.

scholarly journals Resistência antimicrobiana e perfil plasmidial de Escherichia coli isolada de frangos de corte e poedeiras comerciais no Estado de Pernambuco

2012 ◽  
Vol 32 (5) ◽  
pp. 405-410 ◽  
Author(s):  
Mércia R. Barros ◽  
Wanderley D, da Silveira ◽  
Janete M. de Araújo ◽  
Elizianne P. Costa ◽  
Andrea Alice da F. Oliveira ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Avian Pathogenic Escherichia Coli ◽  
E Coli ◽  
Pathogenic Escherichia Coli

Embora existam linhagens de Escherichia coli não patogênicas para aves, muitas outras possuem a capacidade de causar sérios danos à saúde das mesmas, sendo capazes de ocasionar diferentes tipos de processos infecciosos. As linhagens patogênicas são denominadas Avian Pathogenic Escherichia coli (APEC), possuindo genes relacionados ao processo de patogênese em epissomos (plasmídios) ou no cromossomo. A presença de plasmídios, contendo genes de resistência a antibióticos em linhagens aviárias, patogênicas ou não, indicam a possibilidade de transferência gênica lateral entre diferentes tipos de linhagens facilitando também a transferência de genes de patogenicidade ou virulência. Objetivou-se com este estudo avaliar o perfil de sensibilidade a antibióticos (13) de diferentes amostras (35) de E. coli isoladas de aves comerciais do Estado de Pernambuco apresentando, ou não, sinais clínicos de processos infecciosos e correlacionar esta resistência com a presença de plasmídios. Os testes utilizados demonstraram que 94,28% dos isolados foram resistentes a três ou mais antibióticos, com a lincomicina apresentando o maior percentual de resistência (100%). Na Concentração Inibitória Mínima (CIM) observou-se multirresistência a vários antimicrobianos. A presença de plasmídios foi detecada em 80,0% (28/35) dos isolados, com 16 isolados apresentando plasmídios com peso molecular aproximado de 88 MDa. Também foi verificada a presença de linhagens apresentando plasmídios de vários tamanhos. Concluiu-se que isolados de E. coli resistentes a antimicrobianos utilizados na avicultura estão presentes no Estado de Pernambuco, tanto em frangos de corte quanto em poedeiras comerciais. A presença de plasmídios detectados na maioria dos isolados pode estar associada à resistência aos antimicrobianos e sugere a presença de possíveis genes relacionados à patogenicidade. Monitorar a resistência a antibióticos em bactérias isoladas de animais torna-se um fator determinante para eleição e êxito do tratamento, bem como a possibilidade de eliminação daquelas que possuem plasmídios para se evitar a transferência de genes relacionados à patogenicidade.

Sign in / Sign up

Export Citation Format

Share Document