Abstract
Ovine footrot is a highly prevalent bacterial disease caused by Dichelobacter nodosus and characterised by the separation of the hoof horn from the underlying skin. The role of innate immune molecules and other bacterial communities in the development of footrot lesions remains unclear. This study shows a significant association between the high expression of IL1β and high D. nodosus load in footrot samples. Investigation of the microbial population identified distinct bacterial populations in the different disease stages and also depending on the level of inflammation. Treponema (34%), Mycoplasma (29%) and Porphyromonas (15%) were the most abundant genera associated with high levels of inflammation in footrot. In contrast, Acinetobacter (25%), Corynebacteria (17%) and Flavobacterium (17%) were the most abundant genera associated with high levels of inflammation in healthy feet. This demonstrates for the first time there is a distinct microbial community associated with footrot and high cytokine expression.
Highlights
Ovine footrot is a highly prevalent bacterial disease caused by Dichelobacter nodosus and characterised by the separation of the hoof horn from the underlying skin
We included a co-regulation analysis. mRNA expression levels of ACTB, Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH), TUBA, PPIA, 18S rRNA, Transmembrane protein 79 (TMEM79), Activating signal cointegrator complex subunit 2 (ASCC2), chromosome 3 open reading frame 58 (C3ORF58), BHLHE40 and DEAD (Asp-Glu-Ala-Asp) box polypeptide 54 (DDX54) were examined in all 40 biopsies by RT-qPCR to verify their reliability as potential reference genes
Comprehensive analysis identified that ACTB, DDX54 and PPIA were the most stably expressed genes and TUBA, 18S rRNA and GAPDH were ranked as the least stably expressed genes (Supplementary Table 2)
Summary
Ovine footrot is a highly prevalent bacterial disease caused by Dichelobacter nodosus and characterised by the separation of the hoof horn from the underlying skin. Fusobacterium necrophorum, a Gram negative bacterium, is attributed to play a role in the initiation of ID prior to D. nodosus infection[1], and/or in the exacerbation of footrot lesions once it is established[5] Other bacteria such as Treponema have been detected in footrot cases, but its role in the disease process remains unclear[6,7,8]. Following experimental infection with D. nodosus and vaccination no changes in TLR4 mRNA expression in peripheral blood leukocytes was observed; in contrast, TLR2 mRNA expression levels increased in peripheral blood in leukocytes response to both, infection or vaccination[27] In this context, we hypothesise that the pathology of footrot is a host mediated expression of local immune responses, in association with bacterial colonisation, leading to severe inflammation that can progress to underrunning footrot. This study aimed to investigate whether high virulent D. nodosus load and abundance of bacterial populations are associated with high levels of inflammation in the ovine interdigital skin
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