# Genetic Parameters, Linear Associations, and Genome-Wide Association Study for Endotoxin-Induced Cortisol Response in Holstein heifers

## TL;DR

This study explores how Holstein heifers genetically respond to stress from bacterial endotoxin, identifying genes and traits linked to cortisol production.

## Contribution

The study identifies 34 genomic regions and 11 candidate genes associated with cortisol response to endotoxin in Holstein heifers.

## Key findings

- Cortisol response to LPS challenge has moderate heritability (h2 = 0.26) and significant additive genetic variance.
- Candidate genes like CDH2, PARD3, and CFH are linked to immune function and cortisol regulation.
- Two key pathways—immune activation and pituitary development—are central to cortisol production.

## Abstract

This study investigated the genetic basis of cortisol response to immune stress induced by bacterial endotoxin in 252 Holstein heifers. Cortisol response showed significant additive genetic variance, along with moderate heritability (h2 = 0.26) and significant but weak linear associations with key traits, including milk yield, protein yield, and cystic ovaries. A genome-wide association study revealed 34 critical genomic regions and 11 candidate genes, notably CDH2, PARD3, and CFH, which are linked to immune function and hormone regulation. Two key biological pathways—immune activation and pituitary development—were identified as central to cortisol production. These results underscore the potential for incorporating stress-resilience traits into dairy cattle breeding programs.

Lipopolysaccharide (LPS) endotoxin is a well-characterized microbe-associated molecular pattern (MAMP) that forms the outer membrane of both pathogenic and commensal Gram-negative bacteria. It plays a crucial role in triggering inflammatory disorders such as mastitis, acidosis, and septicemia. In heifers, an LPS challenge induces a dynamic stress response, marked by elevated cortisol levels, increased body temperature, and altered immune function. Research indicates that LPS administration leads to a significant rise in cortisol post-challenge. Building on this understanding, the present study aimed to estimate genetic parameters for serum cortisol response to LPS challenge in Holstein heifers and its linear associations with production, health, reproduction, and conformation traits. Additionally, a genome-wide association study (GWAS) was conducted to identify genetic regions associated with cortisol response. A total of 252 animals were evaluated for cortisol response, with correlations estimated between cortisol levels and 55 genomic breeding values for key traits. Genetic parameters and heritability for cortisol response were estimated using Residual Maximum Likelihood (REML) in the Blupf90+ v 2.57 software. Single-Step GWAS (ssGWAS) employing a 10-SNP window approach and 42,123 SNP markers was performed to identify genomic regions that explained at least 0.5% of additive genetic variance. Finally, candidate genes and QTLs located 50 kb up and downstream of those windows were identified. The cortisol response showed significant but weak linear associations with cystic ovaries, body maintenance requirements, lactation persistency, milk yield, and protein yield (p-value ≤ 0.05) and showed suggestive weak linear associations with udder texture, clinical ketosis, heel horn erosion, and milking speed (p-value ≤ 0.15). Cortisol response showed significant additive genetic variance, along with moderate heritability of 0.26 (±0.19). A total of 34 windows explained at least 0.5% of additive genetic variance, and 75 QTLs and 11 candidate genes, comprising the genes CCL20, DAW1, CSMD2, HMGB4, B3GAT2, PARD3, bta-mir-2285aw, CFH, CDH2, ENSBTAG00000052242, and ENSBTAG00000050498, were identified. The functional enrichment analysis allowed us to infer two instances where these gene products could interfere with cortisol production: the first instance is related to the complement system, and the second one is related to the EMT (Epithelium–Mesenchymal Transition) and pituitary gland formation. Among the QTLs, 13 were enriched in the dataset, corresponding to traits related to milk (potassium content), the exterior (udder traits, teat placement, foot angle, rear leg placement, and feet and leg conformation), production (length of productive life, net merit, and type), and reproduction (stillbirth and calving ease). In summary, the cortisol response to LPS challenge in Holstein heifers seems to be moderately heritable and has weak but significant linear associations with important production and health traits. Several candidate genes identified could perform important roles, in at least two ways, for cortisol production, and QTLs were identified close to regions of the genome that explained a significant amount of additive genetic variance for cortisol response. Therefore, further investigations are warranted to validate these findings with a larger dataset.