Abstract: An in vitro method for predicting the biological age of a subject includes: (a) bisulfite treatment of genomic DNA extracted from a biological sample obtained from the subject; (b) amplifying the bisulfite treated genomic DNA from step (a) with specific primers that target at least two Low Methylated Region (LMR) in the genomic DNA from (a) to obtain at least one PCR product; (c) measuring the methylation level of CpG sites in the PCR product of step (b); and (d) determining the biological age of the subject with a statistical prediction algorithm, wherein the statistical prediction algorithm is applied to the measured methylation level to determine the epigenetic age of the subject; wherein the subject is from the Galliformes family; and wherein at least three LMRs are amplified in step (b); and with a proviso that the CpG sites associated with single nucleotide polymorphisms are not considered.

Abstract: The invention provides a method for classifying the gut inflammation status of an avian subject or of a group of avian subjects to be tested, the method comprising the comparison of the average methylation levels within a panel of pre-selected LMRs in the genomic DNA isolated from gut sample material deriving from the individual avian subject or of the group of avian subjects to be tested with the average methylation levels of the same panel of LMRs in the genomic DNA pertaining to one or more reference samples having a negative gut inflammation status.

Abstract: The invention pertains to an in vitro method for predicting the chronological age of healthy Galliformes, the method comprising the steps of: (a.) obtaining genomic DNA from biological sample material deriving from the Galliformes subject or from the Galliformes population to be tested, (b.) determining the methylation level of a set of specific CpG sites in the genomic Galliformes DNA obtained in step (a.), and (c.

Abstract: A computer-implemented method of establishing an epigenetic clock for an avian species including: (a) identifying and determining methylation levels of specific CpG sites within a genomic DNA obtained from a plurality of different biological sample materials deriving from the avian species and representing specific time points within a chronological lifespan of the avian species, (b) excluding all CpG sites associated with single nucleotide polymorphisms (SNPs) from the CpG sites identified in (a), (c) excluding all CpG sites located on sex chromosomes from the CpG sites obtained in (b), (d) performing a tissue-specific normalization for the CpG sites obtained in (c), and (e) correlating the CpG methylation levels of the CpG sites obtained in (d) with chronological age with a penalized regression model.

Abstract: The invention provides a method of establishing a chicken methylation clock comprising: (a) determining the methylation ratio and the read coverage of the genomic CpG sites of an age-correlated training sample of a specific chicken tissue; (b) defining a set of CpG sites having reliable methylation ratios in all training samples of step (a) using a cutoff value; and (c) performing a penalized regression using the methylation ratios of step (b) as input and the age correlated to the training sample as dependent variable, by applying a penalized regression model; thereby obtaining a set of CpG sites with corresponding weighting factors and intercept of the linear model equation as parameters defining the chicken methylation clock.