Department of Animal Sciences, Faculty of Agriculture, Food and Environment, The Hebrew University, Rehovot, Israel.
Theriogenology. 2011 Sep 15;76(5):942-53. doi: 10.1016/j.theriogenology.2011.05.001. Epub 2011 Jun 25.
This study compared immediate and carryover effects of mastitis induced by Gram-negative endotoxin (E. coli LPS) and Gram-positive exosecretions (Staph. aureus ex.) on preovulatory follicle function. Synchronized, uninfected cyclic lactating Holstein cows were treated with PGF(2α) on day 6 of the cycle and 36 h later, a dose of either E. coli LPS (n = 8), S. aureus ex. (n = 10), or saline (n = 9) was administered into the mammary gland. Follicular fluids and granulosa cells were aspirated 6 h later from the preovulatory follicles and cows were treated with GnRH. This (cycle 1; immediate effect) was repeated three times (excluding the mammary injections) to induce three 7 d cycles (cycles 2, 3, and 4; carryover effect). E. coli LPS increased body temperature, plasma cortisol concentration, and somatic cell count (SCC), whereas S. aureus ex. induced a minor, subclinical elevation of SCC and slight rise (NS) in body temperature and cortisol concentration. Follicular estradiol, androstenedione, and progesterone concentrations in the E. coli LPS group decreased (P < 0.05) in cycle 1 to about 40%, 13%, and 35%, respectively, of control levels, whereas in the S. aureus ex. group, only estradiol decreased (P < 0.05), to 56% of control concentrations. In cycles 3 and 4, follicular steroids in the E. coli LPS group returned to control concentrations, whereas in the S. aureus ex. group, follicular concentrations of estradiol and androstenedione were lower (P < 0.10) than in controls. In the control group, the concentrations of all follicular and circulating steroids remained stable (P > 0.05) throughout the study. Follicle size was similar in all groups, but the S. aureus ex. treatment caused a decrease (P < 0.02) in the number of follicles developed in cycles 3 and 4. The mRNA expression of steroidogenic genes and LHCGR in the granulosa cells was not affected (P > 0.05) by either treatment during the study, except for a tendency toward lower (P < 0.1) expression in cycle 1 and lower (P < 0.05) expression in cycle 4 of the latter in the S. aureus ex. group. Strain levels, such as SCC and body temperature, following toxin injection correlated well with the magnitude of the immediate decline in follicular steroids. As is typical for Gram-negative clinical events, E. coli LPS-induced acute mastitis caused immediate, short-term, but not long-term impairment of follicular responses, whereas the Gram-positive S. aureus ex.-induced subclinical mastitis exhibited both immediate and carryover disruptive effects on preovulatory follicle function.
本研究比较了革兰氏阴性内毒素(大肠杆菌 LPS)和革兰氏阳性外分泌物(金黄色葡萄球菌 ex.)诱导的乳腺炎对排卵前卵泡功能的即时和后续影响。同步、未感染的周期性泌乳荷斯坦奶牛在周期的第 6 天用 PGF(2α)处理,36 小时后,将大肠杆菌 LPS(n = 8)、金黄色葡萄球菌 ex.(n = 10)或生理盐水(n = 9)剂量注入乳房。6 小时后从排卵前卵泡中抽吸卵泡液和颗粒细胞,并用 GnRH 处理奶牛。这(第 1 周期;即时效应)重复了三次(不包括乳房注射),以诱导三个 7 天周期(第 2、3 和 4 周期;后续效应)。大肠杆菌 LPS 升高了体温、血浆皮质醇浓度和体细胞计数(SCC),而金黄色葡萄球菌 ex. 则引起 SCC 的轻微亚临床升高和体温和皮质醇浓度的轻微升高(无统计学意义)。大肠杆菌 LPS 组第 1 周期的卵泡雌二醇、雄烯二酮和孕酮浓度分别下降(P < 0.05)至对照水平的约 40%、13%和 35%,而金黄色葡萄球菌 ex. 组仅雌二醇下降(P < 0.05),至对照浓度的 56%。在第 3 和第 4 周期,大肠杆菌 LPS 组的卵泡类固醇恢复到对照浓度,而金黄色葡萄球菌 ex. 组的雌二醇和雄烯二酮的卵泡浓度较低(P < 0.10)。在对照组中,所有卵泡和循环类固醇的浓度在整个研究中保持稳定(P > 0.05)。所有组的卵泡大小相似,但金黄色葡萄球菌 ex. 处理导致第 3 和第 4 周期发育的卵泡数量减少(P < 0.02)。在研究期间,两种处理均未影响(P > 0.05)颗粒细胞中甾体生成基因和 LHCGR 的 mRNA 表达,除了金黄色葡萄球菌 ex. 组在第 1 周期和第 4 周期中存在表达趋势下降(P < 0.1)和表达下降(P < 0.05)。毒素注射后的菌株水平,如 SCC 和体温,与卵泡类固醇的即时下降幅度密切相关。与典型的革兰氏阴性临床事件一样,大肠杆菌 LPS 诱导的急性乳腺炎引起了排卵前卵泡反应的即时、短期但非长期损害,而革兰氏阳性金黄色葡萄球菌 ex.-诱导的亚临床乳腺炎对排卵前卵泡功能既有即时又有后续的破坏作用。