Effects of oral administration of lipopolysaccharide derived from Pantoea agglomerans on innate immunity of mammary glands in dairy goats.

This study aimed to evaluate the effect of orally administered lipopolysaccharide (LPS) derived from Pantoea agglomerans (LPSpa) on innate immune functions, including the concentrations of antimicrobial components and interleukin (IL)-10 in goat milk, for the prevention of goat mastitis. Twelve Tokara goats were divided into two groups of six goats. Goats in the LPSpa and control groups were orally administrated with 0.4 g/kg dextrin with or without 0.02 mg/kg LPSpa for 7 days (day 0-6), respectively. After treatment (i.e., day 7), 1 µg LPS from Escherichia coli O111 (LPSec) was infused into one side of the udder in both groups to induce mastitis. Milk from all sides of the udder, saliva, and feces were collected on days 0 and 7. After LPSec infusion into the udders, milk was collected from the infused side of the udder on days 8, 10, and 12. Milk yields and somatic cell counts were recorded during the examination period. The concentrations of immunoglobulin (Ig) A in saliva, feces, and milk and the concentrations of lactoferrin, goat ß defensin-1 (GBD1), S100A7, and IL-10 in milk were measured. After LPSpa oral administration, the concentrations of GBD-1 and IL-10 in the milk of the LPSpa group were significantly higher on day 7 than those in the control group, and the concentration of IgA in the feces tended to be higher than that in the control group. After LPSec intramammary infusion, S100A7 concentration on day 12 was significantly lower in the LPSpa group than in the control group. These findings suggest that the oral administration of LPSpa may prevent mastitis by increasing the concentration of GBD1 in milk.

Effect of 6-n-propyl-2-thiouracil or dexamethasone administration on the responses of antimicrobial components in goat milk to intramammary lipopolysaccharide infusion.

Heat stress impacts the immune system of dairy animals by altering the hypothalamic-pituitary-adrenal axis and thyroid function, leading to conditions such as hypothyroidism and hypercortisolism. This study aimed to elucidate the effect of hypothyroidism and hypercortisolism on the response of mammary innate immune function to inflammation caused by Escherichia coli in dairy goats. To induce hypothyroidism and hypercortisolism, we administered 6-n-propyl-2-thiouracil (PTU; for 21 days) and dexamethasone (DEX; for 5 days), respectively, to six goats each; six goats without treatment were used as the control group. After treatment, lipopolysaccharide (LPS) from E. coli O111 was infused into the mammary gland. Somatic cell counts (SCC) and levels of lactoferrin (LF), S100A7, immunoglobulin A (IgA), and interleukin-8 (IL-8) in milk until 7 days after LPS infusion were measured. An increase in SCC after LPS infusion was inhibited in both PTU and DEX groups, and an increase in LF after LPS infusion was inhibited in PTU group, compared with that in the control group. The results of the present study suggest that the recruitment of neutrophils and LF production decreased under hypothyroidism or hypercortisolism, which may be one of the causes underlying increased incidence of mastitis in dairy animals under heat stress conditions.

Effect of temporary cessation of milking and estradiol combination on the antimicrobial components in goat milk.

A temporary cessation of milking is widely used in Japan to treat mastitis in dairy cows. Exogenous administration of estradiol (E2) is known to inhibit milk production in dairy cows. Therefore, we aimed to evaluate the effects of the temporary cessation of milking in combination with E2 administration on the antimicrobial components of goat milk. Twelve goats, divided into two groups-with and without E2 injection (E2 and control group, respectively), were subjected to cessation of milking in both udder halves for 3 d (day 0-2). Milk yield in the E2 group was significantly lower than that in the control group on days 7 to 10. The concentrations of cathelicidin-2, IgA, and lactoferrin in the E2 group were significantly higher than those in the control group. These results suggest that the temporary cessation of milking with simultaneous E2 administration leads to a higher concentration of certain antimicrobial components in milk than that observed after using cessation of milking alone. Thus, this combination may contribute to a stronger innate immune system and a faster recovery from mastitis, and might prove to be an alternative to antibiotic treatment upon further research.

Effect of temporary cessation of milking on the innate immune components in goat milk.

Temporary cessation of milking is widely used during the dry period of dairy cows. Temporary cessation of milking induces an increase in the somatic cell count (SCC) and level of several inflammatory components of milk, which is believed to be a local adaptation and defense mechanism of the mammary gland. In Japan, temporary cessation of milking combined with antibiotic administration is widely used to treat mastitis. The present study aimed to elucidate the role of the innate immune system during temporary cessation of milking in a goat model by investigating the concentration of several innate immune components in milk during and around the temporary cessation. In experiment 1, 6 goats were subjected to cessation of milking for 3 d in both udder halves, whereas in experiment 2, 6 other goats were subjected to cessation of milking for 3 d only in 1 udder half. In experiment 1, the milk yield was lower on d 5 and 6, whereas the mean SCC was higher on d 5 compared with d 0 before temporary milking cessation. The concentrations of goat DEFB1, S100A7, cathelicidin-2 and 7 (CATHL-2 and 7), IgA, and lactoferrin were increased after temporary cessation of milking. In experiment 2, the milk yield was lower between d 5 and 7, whereas the mean SCC was higher between d 4 and 7 compared with d 0. The concentrations of CATHL-2, IgA, and lactoferrin were increased after temporary cessation of milking only in the udder half subjected to milking cessation. These results suggest that temporary cessation of milking increase the SCC and concentration of several innate immune components in milk without infection, which may contribute to mastitis treatment.

Association of endometritis and ovarian follicular cyst with mastitis in dairy cows.

The occurrence of multiple metabolic and inflammatory diseases in dairy cows is higher during the periparturient period, which may be triggered by bacterial components, but not a viable bacterium. This study aimed to determine the association of endometritis and ovarian follicular cyst (OFC) with mastitis in dairy cows. Ninety-eight Holstein dairy cows were clinically examined for endometritis and OFC approximately 30-50 days after calving. Blood and milk samples were collected for the determination of milk somatic cell count (SCC); milk interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNFα), and interleukin-8 (IL-8) concentrations; and plasma haptoglobin (Hp) and lipopolysaccharide-binding protein (LBP) concentrations. Of the 98 dairy cows included in this study, 12 were diagnosed with endometritis and 37 cows were identified as OFC-positive, whereas the remaining 49 cows were healthy (without endometritis or OFC). The average and maximum SCCs and plasma Hp and LBP concentrations were not significantly different between the healthy cows and those with endometritis or OFC. However, when the maximum SCC was classified as <300, 300-1,000, or >1,000 × 103 cells/ml, the percentage of cows with the maximum SCC <300 × 103 cells/ml was significantly lower in the endometritis and OFC-positive groups than in the healthy group. These results suggested that cows with endometritis and OFC during the postpartum period exhibit high SCC, indicating that some bacterial components can be transferred between organs.

Translocation of intrauterine-infused bacterial lipopolysaccharides to the mammary gland in dexamethasone-treated goats.

Our previous study showed that intrauterine-infused lipopolysaccharide (LPS) can be translocated to the mammary gland to induce weak inflammation. This study aimed to determine whether dexamethasone treatment facilitated the translocation of LPS from the uterus to the mammary gland to induce a heavy inflammatory response. Sixteen goats were divided into control and LPS groups, subjected to daily dexamethasone administration before saline or LPS infusion. Milk and blood samples were collected before and after LPS infusion to determine the milk yield and somatic cell count (SCC) and blood leucocyte count (BLC), cytokines, antimicrobial peptides and serum amyloid A (SAA) concentrations. Mammary gland tissues were collected from two goats before and 24 hr after LPS infusion for immunohistochemical analysis of LPS. The mean SCC in the LPS group was significantly higher, whereas the milk yield was significantly lower than that in the control group after LPS infusion. The mean BLC in the LPS group was significantly lower than in the control group after LPS infusion. Furthermore, milk concentrations of IL-1ß, S100A8 and lactoferrin were higher in the LPS group than in the control group after infusion. LPS was detected in the connective tissues and inner alveolar spaces of the mammary glands 24 hr after LPS infusion. We concluded that dexamethasone administration facilitated the translocation of intrauterine-infused LPS to the mammary gland, where it induced an inflammatory response. Therefore, LPS translocated from other organs, such as the uterus, can induce heavy inflammation in the mammary gland under immunosuppressive conditions.

Effects of intrauterine infusion of bacterial lipopolysaccharides on the mammary gland inflammatory response in goats.

This study aimed to determine if intrauterine-infused lipopolysaccharides (LPS) can be translocated to the mammary glands and induce an inflammatory response. Thirty-seven goats were divided into two experiments. Nineteen goats (control group, n = 9; LPS group, n = 10) were subjected to intravenous injection of LPS, and eighteen goats (control group, n = 8; LPS group, n = 10) were subjected to intrauterine infusion of LPS. Milk and blood samples were collected before and after the LPS challenge, to measure the blood leukocyte count (BLC), plasma LPS-binding protein (LBP), milk yield, milk somatic cell count (SCC), lactoferrin (LF), milk lactoperoxidase (LPO) activity, and pro- and anti-inflammatory cytokines in plasma and milk. Mammary gland tissues were collected from the parenchyma before and after the LPS challenge, for immunohistochemistry of LPS. In the intravenous injection experiment, the BLC (P < 0.001) and milk yield (P = 0.009) were lower, whereas the LF concentration (P < 0.001) and milk LPO activity (P < 0.001) were higher in the LPS group compared to that in the control group. LPS was detected in the mammary gland 3 and 24 h after intravenous injection of LPS. In the intrauterine infusion experiment, the mean concentrations of IL-1ß and IL-6 in milk were higher in the LPS group compared to that in the control group (P = 0.004 and P = 0.017, respectively), whereas there were no changes in milk yield or SCC. LPS was detected in the connective tissues and interepithelial spaces of the alveoli of the mammary glands 24 h after intrauterine infusion of LPS. We conclude that intrauterine-infused LPS can be translocated to the mammary glands from the uterus, however, the amount of translocated LPS might not be enough to induce symptoms of clinical or subclinical mastitis.