Abstract
Bovine mastitis is perplexing the dairy industry since the initiation of intensive dairy farming, which has caused a reduction in the productivity of cows and an escalation in costs. The use of antibiotics causes a series of problems, especially the formation of bacterial antimicrobial resistance. However, there are limited antibiotic-free therapeutic strategies that can effectively relieve bacterial infection of bovine mammary glands. Hence, in this study, we constructed a mammary gland tissue-specific expression vector carrying the antimicrobial peptide of bovine-derived tracheal antimicrobial peptide (TAP) and evaluated it in both primary bovine mammary epithelial cells (pBMECs) and mice. The results showed that the vector driven by the β-lactoglobulin gene (BLG) promoter could efficiently direct the expression of TAP in pBMECs and the mammary gland tissue of mice. In addition, significant antibacterial effects were observed in both in vitro and in vivo experiments when introducing this vector to bovine-associated Staphylococcus aureus-treated pBMECs and mice, respectively. This study demonstrated that the mammary gland tissue-specific expression vector could be used to introduce antimicrobial peptide both in in vitro and in vivo and will provide a new therapeutic strategy in the treatment of bovine mastitis.
Highlights
Cow mastitis is one of the most prevalent domestic animal diseases and leads to reduced milk production, increased veterinary cost, and early culling [1,2,3]
The bovinederived tracheal antimicrobial peptide (TAP) and by the β-lactoglobulin gene (BLG) promoter genes were cloned from the cDNA and DNA extracted from the whole blood and primary bovine mammary epithelial cells (pBMECs) of a Holstein cow, respectively. pBLG-EGFP-N1 was generated by replacing the AseI–NheI fragment with the BLG promoter using homologous recombination (Thermo Fisher, Shanghai, China)
To express the TAP gene in bovine mammary glands, we selected the promoter of BLG gene, which is expressed in the mammary gland
Summary
Cow mastitis is one of the most prevalent domestic animal diseases and leads to reduced milk production, increased veterinary cost, and early culling [1,2,3]. Tracheal Antimicrobial Peptides for Mastitis studies to find molecular markers through epigenetic and genome sequencing are attempting to improve cow resistance to mastitis [6]. Cow breeds from the mastitis resistance breeding procedure can hardly meet all economic parameters perfectly, such as a decrease in milk production [6]. Mastitis is a lasting and difficult problem that still challenges the global dairy industry
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