An Optimized Protocol for Molecular Screening of Avian Pathogenic Escherichia Coli From Broiler Chickens in South East Queensland, Australia

Awawdeh, Leena; Turni, Conny; Henning, Joerg; Allavena, Rachel; Cobbold, RN; Mollinger, Joanne L.; Gibson, Justine S.

doi:10.3382/japr/pfz078

Cited by 6 publications

(3 citation statements)

References 34 publications

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“…Thus, the involvement of a particular O serotype in the infection process appeared to vary according to the geographical region. Some of the studies conducted in Egypt, nearly isolated the same serotypes with a predominance of the O78 serotype (Awawdeh et al, 2019;Ramadan et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

Investigation of Antibiotic Resistance Pattern and Virulence Determinants in Avian Pathogenic Escherichia coli Isolated from Broiler Chickens in Egypt

Hamed¹,

Elenbaawy²,

Mahmoud³

et al. 2023

WVJ

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Besides its zoonotic importance, avian pathogenic Escherichia coli (APEC) causes substantial financial losses in the poultry industry globally. The progress of antimicrobial resistance in APEC is mainly associated with excessive antimicrobial use and improper sanitation. Since its beginning in the 1970s, the VITEK system has developed into the VITEK 2 system, which has used an automated system to perform all the steps required for microbial identification and antibiotic susceptibility rapidly and accurately. The present study aimed to update the available circulating data about APEC isolates by phenotypic identification, sero-grouping of APEC from broilers chickens and breeders in five governates of Egypt, investigation of their antibiotic resistance pattern by VITEK 2 system, and molecular identification of their virulence determinants. The prevalence of APEC isolated from the different internal organs (liver, lung, heart, heart blood, and spleen) was 67.5%. The most prevalent serotypes were O125, 0114, O44, O127, O142, and O78. Virulence-associated genes (iutA, fimC, and papC) were detected at rates of 84.4%, 74%, and 54.8%, respectively. The highest resistance was found against ampicillin (100%), trimethoprim-sulfamethoxazole (80%), and ampicillin-sulbactam (78.5%), which indicates that the poultry farms need a surveillance and intervention system with proper accuracy and rapidity to prevent the misuse of antibiotics and APEC outbreaks.

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Section: Discussionmentioning

confidence: 99%

Investigation of Antibiotic Resistance Pattern and Virulence Determinants in Avian Pathogenic Escherichia coli Isolated from Broiler Chickens in Egypt

Hamed¹,

Elenbaawy²,

Mahmoud³

et al. 2023

WVJ

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“…Chelex ® Reagent based methods: Three different trials were conducted using Chelex ® 100 (BioRad) solution-based crude DNA extractions from faecal samples and nematodes following a few protocols and studies [ 2 , 18 ] which were proven effective for other species and at the field level. The modified protocols and the sample amounts utilised are briefly described in Figure 1 .…”

Section: Methodsmentioning

confidence: 99%

Point of care colourimetric and lateral flow LAMP assay for the detection of Haemonchus contortus in ruminant faecal samples

et al. 2021

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In this study, we present an optimised colourimetric and a lateral flow LAMP assay for the detection of Haemonchus contortus in small ruminant faecal samples. Using a previously published LAMP primer set, we made use of commercially available colourimetric LAMP and lateral flow kits and combined this into an optimised diagnostic assay which was then tested on field faecal samples from Eastern and South-Eastern Hungary as well as a pure H. contortus egg faecal sample from Košice, Slovakia. Both assays showed no conflicts in visual detection of the results. Additionally, we modified and tested several centrifuge-free DNA extraction methods and one bead-beating egg lysis DNA extraction method to develop a true point of care protocol, as the source of the starting DNA is the main rate-limiting step in farm-level molecular diagnosis. Out of the various methods trialed, promising results were obtained with the magnetic bead extraction method. Sample solutions from the Fill-FLOTAC® technique were also utilised, which demonstrated that it could be efficiently adapted for field-level egg concentration to extract DNA. This proof of concept study showed that isothermal amplification technologies with a colourimetric detection or when combined with a lateral flow assay could be an important step for a true point of care molecular diagnostic assay for H. contortus.

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“…Similar to other studies, isolates with a >93% similarity in their ERIC profile were assumed to be closely related (a clonal group). 18,19 A cluster was defined as a group of isolates that shared ≥80% similarity in their ERIC-PCR profile patterns.…”

Section: Enterobacterial Repetitive Intergenic Consensus (Eric)-pcrmentioning

confidence: 99%

Virulence‐associated genes in faecal and clinical Escherichia coli isolates cultured from broiler chickens in Australia

Awawdeh,

Forrest,

Turni

et al. 2024

Aust Veterinary J

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A healthy chicken's intestinal flora harbours a rich reservoir of Escherichia coli as part of the commensal microbiota. However, some strains, known as avian pathogenic E. coli (APEC), carry specific virulence genes (VGs) that enable them to invade and cause extraintestinal infections such as avian colibacillosis. Although several VG combinations have been identified, the pathogenic mechanisms associated with APEC are ill‐defined. The current study screened a subset of 88 E. coli isolates selected from 237 pre‐existing isolates obtained from commercial poultry flocks in Australia. The 88 isolates were selected based on their enterobacterial repetitive intergenic consensus (ERIC) and antimicrobial resistance (AMR) profiles and included 29 E. coli isolates cultured from chickens with colibacillosis (referred to as clinical E. coli or CEC) and 59 faecal E. coli (FEC) isolates cultured from clinically healthy chickens. The isolates were screened for the presence of 35 previously reported VGs. Of these, 34 were identified, with iucA not being detected. VGs focG, hlyA and sfa/foc were only detected in FEC isolates. Eight VGs had a prevalence of 90% or above in the CEC isolates. Specifically, astA (100%); feoB (96.6%); iutA, iss, ompT, iroN and hlyF (all 93.1%); and vat (89.7%). The prevalence of these were significantly lower in FEC isolates (astA 79.7%, feoB 77.9%, iutA 52.5%, iss 45.8%, ompT 50.9%, iroN 37.3%, hlyF 50.9% and vat 42.4%). The odds ratios that each of these eight VGs were more likely to be associated with CEC than FEC ranged from 7.8 to 21.9. These eight VGs may be used to better define APEC and diagnostically detect APEC in Australia. Further investigations are needed to identify the roles of these VGs in pathogenicity.

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An Optimized Protocol for Molecular Screening of Avian Pathogenic Escherichia Coli From Broiler Chickens in South East Queensland, Australia

Cited by 6 publications

References 34 publications

Investigation of Antibiotic Resistance Pattern and Virulence Determinants in Avian Pathogenic Escherichia coli Isolated from Broiler Chickens in Egypt

Investigation of Antibiotic Resistance Pattern and Virulence Determinants in Avian Pathogenic Escherichia coli Isolated from Broiler Chickens in Egypt

Point of care colourimetric and lateral flow LAMP assay for the detection of Haemonchus contortus in ruminant faecal samples

Virulence‐associated genes in faecal and clinical Escherichia coli isolates cultured from broiler chickens in Australia

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