Please use this identifier to cite or link to this item: https://hdl.handle.net/1822/83896

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dc.contributor.authorBarros, Maria Margaridapor
dc.contributor.authorCampos, Ana Mariapor
dc.contributor.authorOliveira, Ricardopor
dc.contributor.authorCastro, Joanapor
dc.contributor.authorAlmeida, Gonçalo Nietopor
dc.contributor.authorSilva, Sóniapor
dc.contributor.authorMonteiro, Divanildo Outorpor
dc.contributor.authorAlmeida, Carina Manuela Fernandespor
dc.date.accessioned2023-04-11T15:11:19Z-
dc.date.available2023-04-11T15:11:19Z-
dc.date.issued2023-03-21-
dc.identifier.citationBarros, Maria Margarida; Campos, Ana Maria; Oliveira, Ricardo; Castro, Joana; Almeida, Gonçalo Nieto; Silva, Sónia; Monteiro, Divanildo Outor; Almeida, Carina, Detecting Enterotoxigenic Escherichia coli in animal production: method development and validation. Dare2Change - Innovation-Driven Agrifood Business. No. D&PA 42, Porto, Portugal, March 21, 74-75, 2023.por
dc.identifier.urihttps://hdl.handle.net/1822/83896-
dc.description.abstractSwine enteric colibacillosis is a disease characterized by an intestinal infection caused by the colonization of enterotoxigenic Escherichia coli (ETEC). This infection mostly causes illness or death in neonatal and weaned pigs making it responsible for significant economic losses worldwide [1,2]. Bacterial fimbriae (F4/F5/F6/F18) allow the adhesion of the bacteria to epithelial cells, and when both the immunological systems and the gut microbiota are poorly developed, ETEC colonizes and produces one or more enterotoxins (LT/STa/STb) that can have local and systemic effects [3,4]. Therefore, it is of prime importance to monitor and characterize ETEC in the swine industry to develop mitigation strategies. In this study, we aimed to isolate and characterize ETEC strains from faecal porcine swabs. As such, it was developed a methodology to detect the presence of ETEC and their major virulence factors (toxins/fimbriae). This procedure was divided into two phases, firstly the collected swabs were enriched and then then screened for the presence of genetic determinants of toxins (ST/LT/stx2) by real-time PCR (qPCR). Secondly, the positive enrichments were plated in Tryptone Bile X-glucuronide (TBX) agar and incubated at 37 ºC for 24 hours. Fifty characteristic E. coli colonies were then extensively screened for the presence of toxins (Sta/STb/LT/stx2e) and fimbriae (F4/F5/F6/F18/F41) by multiplex-PCR. The development of both qPCR/multiplex-PCR methods, as well as the optimization of the enrichment step, was done using ETEC controls harbouring the above-mentioned toxins/fimbriae. Nonselective and selective (with novobiocin) enrichment in TSB was performed by using ETEC inoculated faeces samples; with the 24h-selective enrichment providing higher ETEC recovery rates. Optimized qPCR conditions for toxins detection were as follow: 95 ºC for polymerase activation/denaturation, 60 ºC for annealing/extension during 40 cycles, and an internal control (pUC19 DNA) was used in each reaction. Multiplex-PCR was optimized through the conditions, 95 ºC initial denaturation and 35 cycles of 94 ºC denaturation, 60 ºC for annealing and 72 ºC for extension/final extension. In sum, this methodology has the potential to be adopted as a routine technique for the rapid detection of ETEC strains in livestock.por
dc.description.sponsorshipThis work was financially supported by: Project PTDC/CVT-CVT/4620/2021, funded by FEDER funds through COMPETE2020–Programa Operacional Competitividade e Internacionalização (POCI) andby national funds (PIDDAC) through FCT/MCTES; LA/P/0045/2020 (ALiCE), UIDB/00511/2020 and UIDP/00511/2020 (LEPABE), funded by national funds through FCT/MCTES (PIDDAC), and by FCT under the scope of the strategic funding of UIDB/04469/2020 unit (CEB).por
dc.language.isoengpor
dc.relationinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FCVT-CVT%2F4620%2F2021/PTpor
dc.relationLA/P/0045/2020por
dc.relationinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F00511%2F2020/PTpor
dc.relationinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F00511%2F2020/PTpor
dc.relationinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F04469%2F2020/PTpor
dc.rightsopenAccesspor
dc.titleDetecting Enterotoxigenic Escherichia coli in animal production: method development and validationpor
dc.typeconferenceAbstractpor
dc.peerreviewedyespor
dc.relation.publisherversionhttps://dare2change.pt/por
dc.commentsCEB56156por
sdum.event.typeconferencepor
oaire.citationStartPage74por
oaire.citationEndPage75por
oaire.citationIssueD&PA 42-
oaire.citationConferencePlacePorto, Portugalpor
dc.date.updated2023-04-11T09:42:08Z-
dc.description.publicationversioninfo:eu-repo/semantics/publishedVersion-
sdum.conferencePublicationDare2Change - Innovation-Driven Agrifood Businesspor
Appears in Collections:CEB - Resumos em Livros de Atas / Abstracts in Proceedings

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