Effect of intramammary lipopolysaccharide challenge after repeated intrauterine infusion of lipopolysaccharide on the inflammation status of goat mammary glands.

Previous studies have shown that a single infusion of lipopolysaccharide (LPS) into the uterus induces mammary gland inflammation. However, repeated LPS infusions return the mammary glands to their basal state of inflammation. To confirm that this is a state of tolerance to LPS, we examined whether tolerance induced by repeated intrauterine LPS infusions limits mammary gland inflammation following subsequent intramammary LPS infusions. In the first experiment, three goats were treated with repeated intrauterine infusions of LPS dissolved in black ink for 5 consecutive days. Blood and milk samples were collected at 2, 4, 6, 12, 24, 48, 72, 96, and 120 h and smeared on glass slides to confirm the translocation of LPS from the uterus to the mammary gland. Black particles were detected in the blood and milk samples more than 2 h after the first infusion and in the connective tissue of the mammary gland after day 5. In the second experiment, goats were divided into two groups: an intrauterine infusion group (IU; n = 7) and a control group (CON; n = 6). The IU group received an intrauterine infusion of 100 µg of LPS in saline for 5 days. Subsequently, LPS was infused into the mammary glands of both groups to examine the effect of intrauterine treatment on the mammary inflammatory response after intramammary LPS infusion. Blood and milk samples were collected at 6, 12, and 24 h, and then daily until 7 d after the intramammary LPS challenge. Interestingly, a significant increase in the milk somatic cell count (SCC), IL-8, IL-1ß, and TNF-α concentrations were observed in the CON group compared to the IU group. This suggests that pretreatment with repeated intrauterine infusions of LPS suppresses the inflammatory responses to subsequent intramammary LPS challenges.

Effect of repeated intrauterine infusion of lipopolysaccharides on mastitis in goats.

A single infusion of lipopolysaccharide (LPSs) into the uterus induces inflammation in the mammary gland. This indicates that LPS can translocate from the uterus to the mammary gland. Natural endometritis is characterized by continuous intrauterine inflammation. The aim of the present study was to determine the effect of repeated intrauterine infusion of two different types of LPSs obtained from Escherichia coli O111:B4 (LPS-O111) and O55:B5 (LPS-O55) on the inflammatory status of the mammary glands of goats. Goats were assigned to three groups: LPS-O111, LPS-O55, and saline (control). Saline with (LPS-O111 and 55 groups) and without (control) 100 µg LPS was infused into the uterus continuously for 7 days. Decreased milk yield was detected in both LPS-O111 and LPS-O55 groups 2 days after the first LPS infusion. While somatic cell count (SCC) was significantly increased in all groups 1 day after the first LPS infusion, both LPS infusions further increased SCC 2 days after the first infusion and showed a significantly higher SCC than that in the control group. Plasma LPS-binding protein (LBP) was significantly higher in both LPS groups than in the control group during the days after infusion. In addition, pro-inflammatory cytokines, interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α), and IL-8, were significantly increased in both LPS infusion groups compared with those in the control group. The LPS-O111 infusion resulted in higher SCC, LBP, TNF-α, and IL-8 concentrations than those in the LPS-O55 group. These results suggest that repeated LPS infusion into the uterus can induce more severe mammary gland inflammation than a single infusion. Interestingly, the mammary tissues recovered from inflammation even though the LPS intrauterine infusion was continued.