Moyes K M, Drackley J K, Salak-Johnson J L, Morin D E, Hope J C, Loor J J
Mammalian NutriPhysioGenomics, Department of Animal Sciences, University of Illinois, Urbana, Illinois 61801, USA.
J Dairy Sci. 2009 Sep;92(9):4301-16. doi: 10.3168/jds.2009-2170.
Ten multiparous Holstein cows were used to determine the effects of negative energy balance (NEB) on the immune response to a Streptococcus uberis (strain O140J) mastitis challenge during midlactation. Before the study, milk from all quarters of each cow was bacteriologically negative, with a composite somatic cell count of <200,000 cells/mL. Cows were paired based on parity, days in milk, and milk yield. At approximately 77 d in milk, half the cows (n = 5) were feed-restricted to 60% of calculated net energy for lactation requirements to induce NEB. Feed restriction lasted 7 d. Control cows (n = 5) were fed the same diet ad libitum (i.e., positive energy balance; PEB). After 5 d, one rear quarter in all cows was inoculated with 5,000 cfu of Strep. uberis. Jugular blood and aseptic quarter milk samples were collected daily until inoculation and every 6 h postinoculation for 36 h. Blood was analyzed for nonesterified fatty acids, beta-hydroxybutyrate, insulin, cortisol, albumin, serum amyloid A (SAA), and haptoglobin (Hp). Periodically throughout the trial period, blood neutrophils were isolated for determination of cell morphology, chemotaxis, and phagocytosis capability in vitro. Quarter milk samples were analyzed for concentrations of SAA, Hp, cytokines (tumor necrosis factor-alpha, IL-10 and IL-1beta), and activity of respiratory burst enzymes (superoxide dismutase and glutathione peroxidase). All cows developed local and systemic signs of mastitis and calculated NEB was similar to that of cows experiencing postpartal NEB. Serum glucose and insulin concentrations increased in both groups after challenge, most likely because of enhanced glycogenolysis and gluconeogenesis; results indicate that immune cell function may be glucose dependent. Serum cortisol concentration was higher in NEB than PEB cows during feed restriction only (before inoculation), and serum albumin concentration was higher in NEB than PEB cows during the infection period. Compared with PEB, cows in NEB had lower SAA concentrations in serum after 5 d of feed restriction but higher SAA concentrations in milk after Strep. uberis challenge. Serum Hp concentration was higher by 36 h postchallenge in NEB than in PEB cows. Phagocytic capability of neutrophils was lower in NEB than in PEB cows at 0 h of infection but decreased in both PEB and NEB cows through 36 h postinfection. Our results indicate that cows subjected to dietary-induced NEB during midlactation had relatively minimal alterations in immune function.
选用10头经产荷斯坦奶牛,以确定负能量平衡(NEB)对泌乳中期乳房受到乳房链球菌(菌株O140J)感染时免疫反应的影响。在研究开始前,每头奶牛所有乳区的牛奶细菌学检测均为阴性,混合体细胞计数<200,000个细胞/毫升。根据胎次、泌乳天数和产奶量对奶牛进行配对。在泌乳约77天时,将一半奶牛(n = 5)的采食量限制为计算出的泌乳净能量需求的60%,以诱导负能量平衡。限饲持续7天。对照组奶牛(n = 5)自由采食相同日粮(即正能量平衡;PEB)。5天后,给所有奶牛的一个后乳区接种5000 cfu乳房链球菌。每天采集颈静脉血和无菌乳区奶样,直至接种,并在接种后每6小时采集一次,共采集36小时。分析血液中的非酯化脂肪酸、β-羟基丁酸、胰岛素、皮质醇、白蛋白、血清淀粉样蛋白A(SAA)和触珠蛋白(Hp)。在整个试验期间定期分离血液中的中性粒细胞,以测定其体外细胞形态、趋化性和吞噬能力。分析乳区奶样中SAA、Hp、细胞因子(肿瘤坏死因子-α、IL-10和IL-1β)的浓度以及呼吸爆发酶(超氧化物歧化酶和谷胱甘肽过氧化物酶)的活性。所有奶牛均出现乳房炎的局部和全身症状,计算出的负能量平衡与产后负能量平衡奶牛相似。攻毒后两组奶牛的血清葡萄糖和胰岛素浓度均升高,很可能是由于糖原分解和糖异生增强;结果表明免疫细胞功能可能依赖葡萄糖。仅在限饲期间(接种前),负能量平衡组奶牛的血清皮质醇浓度高于正能量平衡组奶牛,在感染期间,负能量平衡组奶牛的血清白蛋白浓度高于正能量平衡组奶牛。与正能量平衡组相比,限饲5天后负能量平衡组奶牛血清中SAA浓度较低,但在乳房链球菌攻毒后牛奶中SAA浓度较高。攻毒后36小时,负能量平衡组奶牛血清Hp浓度高于正能量平衡组奶牛。感染0小时时,负能量平衡组奶牛中性粒细胞的吞噬能力低于正能量平衡组奶牛,但在感染后36小时内,正能量平衡组和负能量平衡组奶牛的吞噬能力均下降。我们的结果表明,泌乳中期因日粮诱导出现负能量平衡的奶牛免疫功能变化相对较小。