高谷物日粮诱导的亚急性瘤胃酸中毒（SARA）会损害反刍动物的瘤胃上皮屏障功能，但SARA是否会持续性损害瘤胃上皮屏障的形态结构和功能还尚不清楚。本研究旨在以泌乳山羊为动物模型，研究SARA是否对瘤胃上皮的形态结构、通透性及参与上皮屏障功能的关键基因表达具有持续性影响。选取12只安装有永久性瘤胃瘘管的泌乳中期奶山羊，随机分为对照组（Ctrl，n=4）和SARA组（n=8），对照组试验动物饲喂NFC/NDF比为1.40的基础饲粮，SARA组依次饲喂NFC/NDF比为1.40、1.79、2.31和3.23四种饲粮诱导试验动物发生SARA。SARA诱导成功后从SARA组中随机选取4只患病动物让其自由采食混合粗饲料4周使其恢复，即post-SARA组。采用pH值监测系统连续监测瘤胃pH值以判定SARA的严重程度。采集瘤胃腹囊上皮组织，利用透射电镜、尤斯灌流室、PCR和Western blot等先进技术检测瘤胃上皮的形态结构和功能。结果表明：与对照组相比，（1）SARA组瘤胃上皮乳头长度、宽度、表面积和角质层厚度显著增加(P<0.05)，棘基层厚度和上皮总厚度则显著降低(P<0.05)。post-SARA组这些参数则趋向于恢复到对照组水平(P>0.05)。同时透射电镜结果显示，SARA减少了瘤胃上皮紧密连接数量，加宽了上皮细胞之间的间隙。（2）SARA组和post-SARA组瘤胃上皮短路电流（Isc）、组织导电性（Gt）以及辣根过物氧化酶（HRP）通过瘤胃上皮的流速均显著增加(P<0.05)，这表明SARA可引起瘤胃上皮通透性持续升高，进而导致其屏障功能长期受损。（3）SARA组和post-SARA组瘤胃上皮紧密连接蛋白CLDN1, OCLN and ZO-1的mRNA和蛋白表达量均极显著下调(P<0.01)。由此可见，SARA可导致瘤胃上皮屏障结构和功能持续受损，这与瘤胃上皮紧密连接蛋白表达下调密切相关，而且瘤胃上皮屏障功能的恢复滞后于形态结构的恢复。

Isolated ruminal epithelia from caudal blind sacs of dairy goats were incubated with butyrate and insulin-like growth factor-1 (IGF-1) at different concentrations. Proportions of ruminal epithelium in different phases of the cell division cycle were determined by flow cytometric analysis. The proportion of epithelial cells in S phase and G2-M phase (PS&G2-M) increased significantly (P<0.01) whereas the proportion of epithelial cells in G0-G1 phase (PG0-G1) decreased after incubation with IGF-1. PS&G2-M decreased whereas PG0-G1 increased markedly (P<0.01) after incubation with sodium butyrate. PS&G2-M incubated with IGF-1 and butyrate sodium together increased more than that incubated with IGF-1 alone; PG0-G1, however, decreased significantly (P<0.01). Our results indicate that IGF-1 enhances whereas sodium butyrate inhibits the proliferation of rumen epithelial cells. Furthermore, butyrate and IGF-1, together, have a synergic effect on the proliferation of rumen epithelium.

The application of n-alkane as markers to estimate herbage intake, apparent digestibility and species composition of diet consumed by grazing sheep was studied. Six local Mongolian sheep were used to determine dry matter (DM) intake, apparent DM digestibility and species composition of diet during summer, autumn and winter. Animals were orally dosed twice daily with n-alkane gelatin capsules containing 60 mg C32-alkane as an external marker. Diet composition was estimated by comparing the odd-chain n-alkanes pattern profile (C27-C31) of the consumed plant species with the n-alkanes fecal concentrations of grazing animals, using a non-negative least squares algorithm called EATWHAT software package. The alkane pair C32:C33 and C33 alkane were used to estimate DM intake and diet apparent DM digestibility, respectively. The results showed that daily dry matter intake of the sheep were 1.77, 1.61 and 1.18 kg d-1 in summer, autumn and winter, respectively. Apparent DM digestibility, crude protein (CP), metabolizable energy (ME) and neutral detergent fiber (NDF) intake of diet consumed by sheep decreased significantly (P<0.01) from summer to winter, with no evident changes in ADF and ADL intake. Diet composition indicated Artemisia frigida Willd was the most dominant diet component, contributed 79.68, 68.12 and 86.26% of sheep’s diets in summer, autumn and winter, respectively. Cleistogenes songorica Ohwi and Convolvulus ammannii Desr were the important components of the diet. Although Stipa breviflora Griseb is one of the main plant species in the study area, the sheep rarely choosed it. The study indicated that CP and ME in diet consumed by sheep were deficient in winter. Therefore, appropriate supplementation strategies should be indispensable during this period.

Six rumen-cannulated lactating Guanzhong goats were used to investigate changes in ruminal fermentation pattern and the microbiota following a subacute ruminal acidosis (SARA) inducing procedure. Induction of SARA was performed by increasing dietary non-fiber carbohydrate (NFC) to neutral detergent fiber (NDF) ratio from 1.02 (stage 1) to 1.24 (stage 2), 1.63 (stage 3) and 2.58 (stage 4). A dynamic pH monitoring system, real-time fluorescent quantitative PCR and conventional anaerobic culture were used to assess changes in ruminal pH and microbiota. Results indicated that rumen fermentation patterns changed significantly with increased NFC:NDF ratio. The decline in ruminal pH was caused by increased ruminal total volatile fatty acids (TVFA), which was mainly attributed to a significant increase in ruminal butyrate, rather than the accumulation of ruminal lactic acid. In addition, in the course of SARA, the number of rumen microoganisms altered significantly, with increases in ruminal amylolytic bacteria, Lactobacilli, Streptococcus bovis and Megasphaera elsdenii, the latter particularly dramatically indicating that it may be the main factor responsible for the increase in butyrate, and decrease in protozoa