Neiker Instituto Vasco Of Investigacian Y Desarrollo Agrario

Spain

Neiker Instituto Vasco Of Investigacian Y Desarrollo Agrario

Spain
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Elguezabal N.,Neiker Instituto Vasco Of Investigacian Y Desarrollo Agrario | Chamorro S.,Neiker Instituto Vasco Of Investigacian Y Desarrollo Agrario | Molina E.,Neiker Instituto Vasco Of Investigacian Y Desarrollo Agrario | Garrido J.M.,Neiker Instituto Vasco Of Investigacian Y Desarrollo Agrario | And 3 more authors.
Gut Pathogens | Year: 2012

Background: Inflammatory Bowel Disease (IBD), which includes both Crohns disease (CD) and ulcerative colitis (UC), is caused by a complex interplay involving genetic predisposition, environmental factors and an infectious agent. Mycobacterium avium subsp. paratuberculosis (MAP) is a promising pathogen candidate since it produces a chronic intestinal inflammatory disease in ruminants that resembles CD in humans. MAP is a ubiquitous microorganism, although its presence in the food chain, especially in milk from infected animals, is what made us think that there could be an association between lactase persistence (LP) and IBD. The LCT mutation has brought adaptation to dairy farming which in turn would have increased exposure of the population to infection by MAP. NOD2 gene mutations are highly associated to CD. Methods: In our study, CD and UC patients and controls from the North of Spain were genotyped for the lactase gene (LCT) and for three NOD-2 variants, R702W, G908R and Cins1007fs. MAP PCR was carried out in order to assess MAP infection status and these results were correlated with LCT and NOD2 genotypes. Results: As for LP, no association was found with IBD, although UC patients were less likely to present the T/T -13910 variant compared to controls, showing a higher C-allele frequency and a tendency to lactase non-persistence (LNP). NOD2 mutations were associated to CD being the per-allele risk higher for the Cins1007fs variant. MAP infection was more extended among the healthy controls (45.2%) compared to CD patients (21.38%) and UC patients (19.04%) and this was attributed to therapy. The Asturian CD cohort presented higher levels of MAP prevalence (38.6%) compared to the Basque CD cohort (15.5%), differences also attributed to therapy. No interaction was found between MAP infection and LCT or NOD2 status. Conclusions: We conclude that LP is not significantly associated with IBD, but that MAP infection and NOD2 do show not mutually interacting associations with IBD. © 2012 Elguezabal et al.; licensee BioMed Central Ltd.


Ros-Garcia A.,Neiker Instituto Vasco Of Investigacian Y Desarrollo Agrario | Nicolas A.,Occs Menorca SL | Garcia-Perez A.L.,Neiker Instituto Vasco Of Investigacian Y Desarrollo Agrario | Juste R.A.,Neiker Instituto Vasco Of Investigacian Y Desarrollo Agrario | Hurtado A.,Neiker Instituto Vasco Of Investigacian Y Desarrollo Agrario
Parasites and Vectors | Year: 2012

Background: The tick-borne apicomplexan bovine parasite Theileria annulata is endemic in many tropical and temperate areas, including Minorca (Balearic Islands, Spain). Real-time PCR is widely used for the detection of piroplasms but quantification is not commonly considered.Results: We developed a real-time quantitative PCR (qPCR) assay for the detection and quantification of T. annulata that included an internal amplification control (IAC) to monitor for the presence of potential inhibitors. Specificity, sensitivity, precision, linear range and PCR efficiency were calculated and different methods for transformation of quantification cycle (Cq) values into quantities (Q) were evaluated. The assay was able to detect (100% probability) and quantify (linear response) 100 gene copies, and clinical sensitivity was set at 10 T. annulata per ?l of blood. The assay was then validated on 141 bovine blood samples analyzed in parallel by a LuminexW suspension array, showing the utility of the qPCR assay developed here for the detection and quantification of the parasite in field conditions. Once validated it was used to monitor T. annulata parasitaemia throughout a year in 8 carrier animals from a farm in Minorca. Conclusions: The developed qPCR assay offers a reliable and simple way to quantify T. annulata infection loads, which could prove crucial in studying the role of carrier animals as a source of the infection, or assessing the efficacy of treatment and control measures. © 2012 Ros-Garca et al.; licensee BioMed Central Ltd.

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