人工饲粮对幼年工蜂生长发育及生理代谢的影响

本试验利用液相色谱-质谱联用技术(LC-MS)对幼年工蜂进行代谢组学分析,旨在探究不同人工饲粮对幼年工蜂生长发育及生理代谢的影响,为蜂王邮寄的人工饲粮配制提供一定理论依据。试验选取1日龄的意大利蜜蜂(Apis mellifera ligustica)300只,分为5组,分别饲喂5种人工饲粮,每组30只工蜂。对照组饲粮:蔗糖粉∶蜂蜜=3∶1;试验组A饲粮:蔗糖粉∶蜂粮∶蜂蜜=6∶5∶1;试验组B饲粮:蔗糖粉∶蜂粮∶蜂蜜=6∶3∶1;试验组C饲粮:蔗糖粉∶花粉∶蜂蜜=6∶5∶1;试验组D饲粮:蔗糖粉∶花粉∶蜂蜜=6∶3∶1。用5种人工饲粮进行室内喂养,记录蜜蜂7 d内的死亡数量;解剖试验组A和对照组第1、3、5、7天的工蜂咽下腺,测定其平均重量、颜色和饱满度;采用LC-MS方法检测试验组A和对照组蜜蜂饲喂7 d后的代谢物差异,对检测数据进行模式识别分析,筛选并鉴别差异代谢物。结果表明:1)对照组和试验组A蜜蜂第1~5天时均没有出现死亡,在第6和7天时试验组A蜜蜂死亡数量均显著低于对照组(P<0.05)。试验组B、C、D的蜜蜂在6 d内全部死亡。2)试验组A第7天时的蜜蜂咽下腺平均重量显著高于对照组(P<0.05),咽下腺颜色为全部乳白色,且咽下腺小体饱满。3)采用主成分分析(PCA)、偏最小二乘法判别方法(PLS-DA)和正交偏最小二乘方-判别分析(OPLS-DA)分析代谢组数据,结果显示试验组A和对照组明显分离,共鉴定出23个差异代谢物,包括氨基酸、脂类、糖类等,其中有10种代谢物上调,13种代谢物下调。差异代谢物通路分析发现,差异极显著的通路有牛磺酸和亚牛磺酸生物合成通路、赖氨酸降解通路、戊糖磷酸盐代谢通路、戊糖和葡萄糖醛酸相互转化通路(P<0.01),差异显著的通路有氨基酸的生物合成通路,色氨酸代谢通路,碳代谢通路,丁酸代谢通路,丙氨酸、天冬氨酸和谷氨酸代谢通路,丙酮酸代谢通路(P<0.05)。由此可见,试验组A的饲粮更适合蜜蜂工蜂的生长发育和生存,在邮寄环境中可代替常用的炼糖饲粮。     

Mammary carcinoma arising in an adenoma in a ewe

A large, firm, multi-cystic mammary gland mass grew slowly over 4 y in a 12-y-old, female Finn–Shetland cross sheep. A diagnosis of epithelial malignancy was suspected following fine-needle aspiration cytology at 30 mo after initial observation. The sheep was euthanized when the flock was downsized 18 mo later. A field postmortem examination revealed a large mammary mass, but an absence of metastases to internal organs. Imprint cytology of the mammary tissue supported a benign proliferative process. Histologically, mammary tissue was obliterated by cystic, tubular, and papillary adenomatous arrangements of mammary epithelium, with an anaplastic component, consistent with mammary carcinoma arising in an adenoma. IHC showed strong nuclear positivity to the antibody against progesterone receptor and minimal positivity to the antibody against estrogen receptor alpha expression. Intrinsic subtyping for basal or luminal epithelial origin was attempted through adaptation of companion animal IHC classification panels; high- and low-molecular-weight cytokeratins (CK5, CK8, CK18) failed to stain, but p63 expression for basal epithelium was positive.     

蜂胶对细菌脂多糖刺激下奶牛乳腺上皮细胞炎症相关基因mRNA转录水平和紧密连接蛋白的影响

本试验旨在研究中国蜂胶乙醇提取物（ethanol extract of Chinese propolis，EECP）对细菌脂多糖（lipopolysaccharide，LPS）刺激下体外培养奶牛乳腺上皮细胞炎症相关基因mRNA转录水平和紧密连接渗透性的影响。EECP中总酚酸和总黄酮含量测定采用福林酚法和硝酸铝法，并建立LPS诱导奶牛乳腺上皮细胞（bovine mammary epithelial cells，MAC-T）炎症模型，采用CCK-8法测定EECP对MAC-T相对增殖率的影响，利用实时荧光定量PCR（RT-qPCR）评估EECP对LPS诱导的MAC-T细胞炎症相关因子（IL-6、IL-8、TNF-α和IL-1β）相对mRNA转录水平；以及对紧密连接蛋白（occludin、ZO-1）相对mRNA转录水平进行检测，并进一步利用免疫荧光技术对紧密连接膜蛋白进行定位，确定EECP对LPS诱导MAC-T细胞炎症紧密连接渗透性的影响。结果显示：EECP中总酚酸含量为106.35 mg没食子酸当量（GAE）·g^-1、总黄酮含量为320.85 mg芦丁当量（RE）·g^-1；CCK-8结果显示EECP的安全浓度为0~15 μg·mL^-1，并可有效提高LPS刺激下MAC-T的活力；LPS刺激显著增加了细胞炎症相关因子IL-6、IL-8、TNF-α和IL-1β mRNA的转录量（P<0.001）；但2.5~15.0 μg·mL^-1 EECP预处理显著降低了IL-6、IL-8、TNF-α和IL-1β mRNA的转录量；与此类似，LPS刺激显著抑制了紧密连接蛋白基因（occludin、ZO-1）mRNA的转录量（P<0.01），而EECP预处理后紧密连接蛋白基因（occludin和ZO-1）mRNA的转录量显著增加（P<0.05）；免疫荧光染色试验也证实EECP能通过上调紧密连接蛋白（occludin、ZO-1）的表达，缓解LPS诱导的乳腺上皮细胞屏障功能紊乱。该结果证实，EECP对细菌脂多糖诱导奶牛乳腺上皮细胞炎症具有良好的保护作用，这为利用中国蜂胶预防奶牛乳腺炎提供了试验基础。     

宽口裂腹鱼消化系统解剖和组织学观察

该研究采用解剖、石蜡切片和HE染色法对宽口裂腹鱼(Schizothorax eurystomus)消化系统解剖特征和组织切片进行观察。结果显示,其消化管管壁由内向外分别为黏膜层、黏膜下层、肌层和浆膜,主要差别在于黏膜层和肌层。食道黏膜上皮为复层扁平上皮,上皮间分布有大量杯状细胞;前肠和中肠黏膜上皮为单层柱状上皮,明显可见刷状缘、杯状细胞和淋巴细胞分布其间;后肠黏膜上皮为假复层柱状上皮,其间也有杯状细胞和淋巴细胞分布,肠道中杯状细胞由前至后逐渐增多。食道肌层为内环外纵的骨骼肌;前肠肌层为内环外纵的平滑肌;中肠和后肠为内螺旋外环行的平滑肌。消化腺由肝脏和胰腺组成,胰腺弥散状分布在肝脏中,肝小叶不明显。研究表明,宽口裂腹鱼消化系统组织学特征与其食性具有适应性。     

Effect of starvation on the performance of baby octopus (Robsonella fontaniana) paralarvae

The aim of this study was to evaluate the effect of short‐ and long‐term starvation on paralarvae from hatching and compare to fed paralarvae. In the continuous starvation treatments, paralarvae at hatching were left without food as independent treatments for 5, 8, 9, 10, 11, 12, 13 and 15 days. In the fed treatments, the newly hatched paralarvae were fed for five and eight days; then each group was left in starvation as independent treatments for 3, 5 and 7 days. After any experimental starvation period, the paralarvae were fed for five more days to evaluate their recovery. Paralarvae exposed to continuous starvation from hatching endured up to 8 days after hatching (8 SDAH) showing significant recovery. Its survival decreased proportionally to the days of starvation, without any recovery after 12 DAH. Fed paralarvae (5 and 8 FDAH), resulted in significant differences accordingly to the length of the starvation period. Paralarvae left under permanent starvation showed a noticeable decrease in their arm/mantle length ratio and an atrophy of the digestive gland was observed. Amino acids were used primarily as energy source by paralarvae, particularly in the second week of starvation. It is discussed the critical age to avoid deleterious effects of starvation on paralarvae.     

通草提取物对奶牛乳腺上皮细胞乳糖合成及相关基因表达的影响

为探讨通草对奶牛乳腺上皮细胞乳糖合成及相关基因表达的影响,采用不同剂量通草提取物处理奶牛乳腺上皮细胞,利用四甲基偶氮唑盐(MTT)法检测乳腺上皮细胞增殖能力,乳糖/半乳糖检测试剂盒(Lactose/D-Galactose (Rapid) Assay Kit)检测乳腺上皮细胞培养液中乳糖含量,实时荧光定量PCR技术检测乳腺上皮细胞乳糖合成相关基因葡萄糖转运蛋白1(GLUT1)、葡萄糖转运蛋白4(GLUT4)、葡萄糖转运蛋白8(GLUT8)、葡萄糖转运蛋白12(GLUT12)、己糖激酶Ⅰ(HKⅠ)、己糖激酶Ⅱ(HKⅡ)、β-1,4-半乳糖基转移酶-1(β-4GALT1)和α-乳清白蛋白(α-LA)基因mRNA表达水平。结果表明,200、400、600μg(生药)·mL-1通草提取物提高奶牛乳腺上皮细胞增殖能力(P<0.05),促进奶牛乳腺上皮细胞合成分泌乳糖(P<0.05),并显著上调细胞GLUT1、GLUT8、HKⅡ、β-4GALT1及α-LA mRNA表达水平(P<0.05),但对GLUT4、GLUT12和HKⅠmRNA表达水平无显著影响(P>0.05)。研究表明,适当浓度通草提取物可提高奶牛乳腺上皮细胞增殖能力,并通过上调乳腺上皮细胞GLUT1、GLUT8、HKⅡ、β-4GALT1和α-LA的mRNA表达,促进乳腺上皮细胞合成乳糖。     

ULTRASONOGRAPHIC EVALUATION OF ADRENAL GLAND SIZE COMPARED TO BODY WEIGHT IN NORMAL DOGS

The accepted cut‐off value for adrenal gland maximum diameter of 0.74 cm to distinguish adrenal gland enlargement in dogs regardless of body weight may not be appropriate for small to medium breed dogs. The purpose of the current retrospective study was to examine adrenal gland dimensions as a function of body weight in healthy dogs in three weight categories (< 10 kg, 10–30 kg, and > 30 kg) representing small, medium, and large breeds, respectively, to establish greater confidence in determining if adrenal gland size is abnormal. The measurements of length (sagittal plane), cranial and caudal pole thickness (sagittal and transverse planes), and caudal pole width (transverse plane) of both adrenal glands were obtained ultrasonographically in clinically healthy dogs (n = 45) with 15 dogs in each weight group. Findings support our hypothesis that adrenal gland size correlates with body weight in normal dogs, and more precise reference intervals should be created for adrenal gland size by categorizing dogs as small, medium, or large breed. The caudal pole thickness of either adrenal gland in a sagittal plane was the best dimension for evaluating adrenal gland size based on low variability, ease, and reliability in measurement.