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Intramammary challenge of lipopolysaccharide stimulates secretion of lingual antimicrobial peptide into milk of dairy cows

N. Isobe^*,1, K. Morimoto, J. Nakamura^*, A. Yamasaki^* and Y. Yoshimura^*

^* Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima 739-8528, Japan
Livestock Technology Research Center, Hiroshima Prefectural Technology Research Institute, Shobara 727-0023, Japan

¹ Corresponding author: niso{at}hiroshima-u.ac.jp

Lingual antimicrobial peptide (LAP) belongs to the β-defensin family in cattle and is found in bovine milk. However, it is unclear whether LAP is involved in the early immune response to mammary infection. The aim of the study was to investigate the changes of LAP concentration in milk after intramammary challenge with lipopolysaccharide (LPS), the gram-negative bacteria cell membrane component, in dairy cows. Milk was collected before and after LPS or phosphate-buffered saline (control) challenge every hour for 12 h on d 0 and twice daily from d 1 to 7. Somatic cell count (SCC), LAP concentration, and lactoperoxidase (LPO) activity in the milk were measured. Somatic cell count started to increase at 2 h postchallenge and remained high until d 5 (694 ± 187 x 10³ to >1,000 ± 0 x 10³ cells/mL at d 0; >1,000 ± 0 x 10³ cells/mL at d 1 to 3; 684 ± 194 x 10³ to 829 ± 108 x 10³ cells/mL at d 4; 527 ± 197 x 10³ to 656 ± 145 x 10³ cells/mL at d 5). Somatic cell count increased in the control cows, although the levels were lower compared with those in the LPS challenge group. The LAP concentration in milk increased significantly at 2 h post-LPS-challenge and was maintained at high levels until d 2 (8.6 ± 0.6 to 17.5 ± 2.3 nM). In the control cow infused with phosphate-buffered saline, there was no increase of LAP concentration in milk (5.1 ± 0.6 to 7.2 ± 0.8 nM). Increase of LPO activity in the milk was observed at 6 h after LPS challenge and continued until d 3 (4.7 ± 0.3 to 9.4 ± 1.1 U). No increase of LPO activity was observed in the milk of control cows. The increase and subsequent decrease in LAP concentration after LPS challenge occurred earlier than those of LPO activity. In multiparous cows with LPS infusion, there was a significantly negative relationship between the days leading to the basal levels in LAP concentration and LPO activity (r = –0.75). These results suggest that LPS induces secretion of LAP into milk within hours and that LPO may have a synergistic antimicrobial function with LAP in mammary glands of dairy cows.

Key Words: lingual antimicrobial peptide • milk • dairy cow • lipopolysaccharide