Relative expression of pro-inflammatory cytokine genes in Holstein dairy cows naturally affected by Escherichia coli mastitis

Document Type : Full paper (Original article)

Authors

1 Graduated from Faculty of Agriculture, University of Yasouj, Yasouj, Iran

2 Department of Animal Science, Faculty of Agriculture, University of Yasouj, Yasouj, Iran

10.22099/ijvr.2025.49388.7250

Abstract

Background: Bovine mastitis, the most common and costly disease, is characterized by mammary gland inflammation. In dairy cattle, Escherichia coli, one of the main causes of mastitis and its lipopolysaccharide (LPS), is a prominent virulence factor. The LPS is responsible for stimulating the expression of pro-inflammatory cytokines that are key components of the early response of the host’s innate immunity and plays an important role in the subsequent inflammatory response to eliminate the infection. Aims: This study aimed to investigate the expression profiles of some pro-inflammatory cytokine genes (IL-6, IL-8, TNF-α, IFN-γ, and GM-CSF) in milk somatic cells of healthy cows and naturally infected cattle with E. coli in their first lactation. Methods: Milk samples were aseptically collected from six healthy cows and six cows with clinical mastitis. In animals with mastitis, those in which the cause of mastitis was only E. coli bacteria were selected for further research. Total RNA was extracted from the somatic cells of milk, the first strand cDNA was synthesized and real-time PCR was performed for the studied genes. As reference genes, β-actin and GAPDH were used to normalize the data. The REST and SAS programs analyzed the real-time data for significance. Results: Pro-inflammatory cytokine genes were expressed in all healthy cows and in cows with clinical mastitis. The expression profiles of cytokine genes showed significantly higher expression of the IL-8, TNF-α, IFN-γ, and GM-CSF (P<0.01) in cows with clinical mastitis compared with animals free of infection. Expression correlations were widely varied between all pairs of genes in healthy animals and those affected by mastitis. In healthy animals, a significant positive correlation was found between the mRNA expression of IL-6 and IL-8 genes (P<0.01). In addition, the mRNA levels of the GM-CFS showed a significant positive correlation with the expressions of both IL-6 and IL-8 genes (P<0.01). In cows suffering from clinical mastitis, an interesting finding was the presence of significant positive correlations between the mRNA levels of the GM-CFS and the expression levels of IL-6, IL-8, IFN-g, and TNF-α genes (P<0.05). Conclusion: The study suggests that the IL-8, GM-CFS, IFN-g, and especially TNF-α genes could be strong indicators of the early response of the immune system in the mammary gland of dairy cows naturally infected by E. coli. However, further studies should be conducted to confirm the findings of this study.

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References

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