Hepatic DNA Methylation in Response to Early Stimulation of Microbiota with Lactobacillus Synbiotics in Broiler Chickens

Abstract

DNA methylation inhibits DNA transcription by the addition of methyl residues to cysteine within the CpG islands of gene promoters. The process of DNA methylation can be modulated by environmental factors such as intestinal microbiota. In poultry, the composition of the intestinal microbiota can be stimulated by in ovo delivery of synbiotics. The present study aims to determine the effect of Lactobacillus synbiotics delivered in ovo on the level of hepatic DNA methylation in broiler chickens. In ovo stimulation was performed on day 12 of egg incubation. Bioactive compounds delivered in ovo included (S1)—Lactobacillus salivarius with GOS and (S2)—Lactobacillus plantarum with RFO. Samples were collected from six individuals from each group on day 42 post-hatching. DNA methylation of five genes selected on the basis of the transcriptome data were analyzed using the qMSP method. Significant changes were observed in DNA methylation of genes in liver including ANGPTL4 and NR4A3, after S2 delivery. The obtained results confirm that the downregulation of metabolic gene expression in the liver mediated by in ovo stimulation had epigenetic characteristics.

Oxidative and Epigenetic Changes and Gut Permeability Response in Early-Treated Chickens with Antibiotic or Probiotic

“…DNA methylation is an epigenetic mechanism that inhibits DNA transcription through the addition of methyl residues to cysteine. DNA methylation can be modulated by environmental factors such as diet, stress, climate, and the gut microbiota [23,[45][46][47]. Literature data show that the level of DNA methylation may be influenced by the short-chain fatty acid butyrate, which is a product of prebiotic fermentation [48,49].…”

“…Literature data show that the level of DNA methylation may be influenced by the short-chain fatty acid butyrate, which is a product of prebiotic fermentation [48,49]. According to Dunisławska et al [23], early stimulation of the gut microbiota of chickens has a strong effect leading to permanent changes in the DNA methylation profile. There is research showing that DNA damage can be reduced by the use of antioxidants.…”

“…In contrast, antibiotics reduce intestinal colonization by both harmful and saprophytic microorganisms [5]. There are reports indicating that intestinal colonization by microorganisms affects protein and DNA oxidation and epigenetic changes in DNA not only in the gut, but in other tissues as well [23]. Negative oxidative and epigenetic changes may result in the induction of intestinal inflammation [24][25][26][27][28], which in turn can directly disturb the integrity of the intestinal barrier [28].…”

Interaction between Early in Ovo Stimulation of the Gut Microbiota and Chicken Host – Splenic Changes in Gene Expression and Methylation

“…Spleen was homogenized with the TissueRuptor homogenizer (Qiagen GmbH, Hilden, Germany) in TRI reagent. Isolation of DNA from spleen was carried out by phenol -chloroform method 38 as described by Dunislawska et al 2020 31 . RNA and DNA quality and quantity was checked by electrophoresis and NanoDrop2000 (Scienti c Nanodrop Products, Wilmington, USA).…”

“…Gene methylation analysis-qMSP DNA methylation is a speci c chemical modi cation of a nucleic acid, involving the attachment of a methylene group to cytosine or adenine nucleotides. The isolated DNA was subjected to methylation analysis using the qMSP method as described by Dunislawska et al 2020 [22]. The qMSP method is a methyl speci c quantitative PCR (qPCR) preceded by DNA conversion using bisul te.…”