基于UHPLC-QTOF-MS代谢组学方法筛选奶牛不同妊娠状态时的候选生物标志物

旨在筛选人工授精后第17天奶牛在不同妊娠状态时的候选生物标志物。本研究以宁夏某奶牛场体重为（550±50） kg,体况评分相近的经产（2~3胎次）健康荷斯坦奶牛为试验对象。同期发情后,在人工授精后第17天晨饲前对奶牛进行尾静脉采血,后期采用计步器和B超仪诊断出奶牛的妊娠状态。根据诊断结果将奶牛分为妊娠组（A组,n=12）和未妊娠返情组（B组,n=24）,将这两组血样进行代谢轮廓及代谢物变化分析。主成分分析（PCA）和正交偏最小二乘法-判别分析（OPLS-DA）结果显示,A、B两组血浆代谢轮廓均发生了明显变化,A、B两组之间共检测出8种ROC （Receiver operating characteristic curve）曲线下面积（area under the ROC curve,AUC）＞0.8的差异代谢物。丙氨酸-亮氨酸、甘氨酸、脯氨酸、L-正亮氨酸、DL-苯丙氨酸、肌氨酸、吡咯-2-羧酸和缬氨酸-蛋氨酸有望成为识别妊娠组（A组）和未妊娠返情组（B组）的差异代谢物。综上表明,血浆中的这8种代谢物有望成为奶牛妊娠识别阶段潜在的生物标志物,为配种早期妊娠识别提供新思路。     

Improving the reproductive,immunity and health status of rabbit does using honey bee venom

The aim of the present study was to investigate the long-term effect of using bee venom (BV) on the reproductive performance, immune, and health status of rabbit does and its effect on their litters. Sixty mature does, from Spanish V-line rabbit stock, were randomly assigned to four homogeneous groups with 15 does each. The 1^st, 2^nd and 3^rd groups were injected twice weekly under the neck skin with 0.1 ml solution contains 0.1, 0.2 and 0.3 mg BV/rabbit respectively. The 4^th group served as a control group. From the results, litter size at birth, litter weight and survival rate at weaning age as well as milk yield were significantly (p ≤ 0.05) increased in BV groups than in the control group. Serum estradiol 17-β (E2) was significantly (p ≤ 0.05) higher (15%) in the rabbit does treated with BV compared to the control group. The treated does with BV at any study doses showed a gradual and significant (p ≤ 0.05) decrease (12%) in serum progesterone levels (P4) compared to the control. They also showed a significant (p≤0.05) increase in conception (17%) and fertility rates (10%) compared to the control does. Treatment of rabbit does with BV caused a gradual and significant (p ≤ 0.05) reduction in both aspartate aminotransferase (AST) (16%) and alanine aminotransferase (ALT) (37%) liver enzyme activities. Additionally, results have shown that BV resulted in a gradual and significant (p ≤ 0.05) increase in total antioxidant capacity (TAC), antioxidative enzymes such as glutathione S-transferase (GST) and glutathione peroxidase (GPx), serum IgG, IgM and IgA levels with significant (p ≤ 0.05) decrease in malondialdehyde (MDA) and thiobarbituric acid reactive substances (TBARS) in BV groups compared to the control group. Results suggest that BV can be used in rabbit farming as an effective and safe alternative to artificial chemical drugs (sexual-stimulants) to improve certain reproductive traits, immune response and health.     

Physiological response of one of South Africa’s premier freshwater sport angling species,the Orange-Vaal smallmouth yellowfish Labeobarbus aeneus,to catch-and-release angling

The practice of catch-and-release fishing has been widely promoted by angling fraternities as a fisheries manage- ment tool. The aim of this investigation was to determine the physiological response of Orange-Vaal smallmouth yellowfish, Labeobarbus aeneus, to catch-and-release angling in the Vaal River, South Africa. Fish were collected using standard fly-fishing techniques, anaesthetised in clove oil and blood was drawn from the caudal vein; thereafter the fish were weighed, measured, revived and released. Blood plasma was analysed for concentra- tions of glucose, cortisol and lactate to determine the effects of angling duration, fish size and water tempera- ture. Larger fish were angled for a longer duration compared with smaller fish. Levels of glucose were affected by water temperature (influenced by time of year). Plasma glucose concentrations decreased with greater angling duration. Few individuals (n = 12) showed increased plasma cortisol concentrations. In extended-capture fish (angled for >1 min), lactate concentrations increased significantly above values for rapid-capture fish (angled for >30 s). These data suggest that catch-and-release causes physiological stress to fish, but nonetheless this practice can be a valuable fisheries management tool to ensure the sustainability of fish populations. Other factors beyond the ‘angling’ time are likely to contribute to physiological disruptions in homeostasis and therefore handling and air exposure of angled fish should be included in future catch-and–release angling studies. In addition, the longer-term impact of angling on fish health should also be determined.     

Effects of formalin on total haemocytes count and histopathological changes in the shrimp Litopenaeus schmitti (Pérez‐Farfante & Kensley 1997)

Formalin is used in shrimp industry to eliminate ectoparasites and as a disinfectant. However, the effect of formalin on shrimp's defense mechanism is unknown. Several biomarkers were used to assess the immune response of juveniles of the shrimp Litopenaeus schmitti exposed to different concentrations of formalin (10, 25 and 50 mg L^?1) during 24 and 48 h. Formalin concentrations of 25 and 50 mg L^?1 produced a significant reduction in the total haemocyte count at both 24 and 48 h. Peroxidase and phenoloxidase activity did not show variations. The total protein concentration in haemolymph of shrimps was not significantly different between experimental groups. Histological analysis showed hyperplasia in the gills of animals exposed at 10 mg L^?1 for 24 h; however, the exposure at 25 and 50 mg L^?1 caused necrosis in gills, hepathopancreas and the antennal gland.     

蚯蚓生物标志在农药污染评价中的应用

农药污染问题越来越多的被人们所重视,单纯依靠化学分析方法来阐释农药污染的危害,已不能真实地判别出农药是否会对环境产生不利影响,因此需要建立有效的生物评价体系来弥补这一缺陷。蚯蚓位于食物链最底层且在土壤生态系统中生物量最大,已被广泛地应用到农药污染状况的评价中,基于分子、细胞水平的蚯蚓生物标志物可以在农药污染的早期阶段就辨别出来。文章在简要回顾了生物标记物发展历史的基础上,从蚯蚓作为土壤生态风险评价的模式生物的特点出发,着重概述了几种蚯蚓生物标志物(乙酰胆碱酶、抗氧化酶系、免疫细胞、基因表达)评价农药污染水平及作用机理的研究进展,并就未来的研究重点和趋势提出见解。     

Melanomacrophages in three species of free-ranging sharks from the northwestern Atlantic, the blue shark Prionacae glauca (L.), the shortfin mako, Isurus oxyrhinchus Rafinesque, and the thresher, Alopias vulpinus (Bonnaterre)

The melanomacrophage aggregates or cells (MMC) are commonly used as biomarkers of exposure to pollution in fish, albeit their numbers and morphological characteristics can be influenced not only by environmental toxins but also by a range of physiological parameters and pathological conditions. Accordingly, before we can use MMC as biomarkers in any fish species, their normal, 'background' characteristics have to be established in apparently healthy fish. The knowledge of MMC in sharks is minimal. The aim of this study was to characterize MMC from 51 free-ranging, large pelagic sharks from the northwestern Atlantic, including shortfin mako, Isurus oxyrhinchus , thresher, Alopias vulpinus and blue shark, Prionacae glauca . The rationale of this study was twofold. First, because, sharks have life histories predisposing them to the accumulation of environmental toxins they constitute good sentinel species of the health of the global marine ecosystem. Second, because many shark populations are in decline or threatened by extinction, we need to expand our understanding of their health status in order to develop appropriate protective measures. All sharks were collected by sports fishing gear between June and August in 2007. Their health condition was assessed by necropsy, morphometrics, and by microscopic examination of gonads, livers, spleens and kidneys. Routine, haematoxylin and eosin and/or Pearl's reagent-stained paraffin embedded sections were studied by light microscopy. Our results provide the first data on the morphometric and morphological characteristics of MMC in viscera of apparently healthy free-ranging sharks from the northwestern Atlantic.     

Stress and pain response after oligofructose induced-lameness in dairy heifers

Lameness is one of the most painful conditions that affects dairy cattle. This study was conducted to evaluate clinical signs and plasma concentration of several pain and stress biomarkers after oligofructose-induced lameness in dairy heifers. Lameness was induced using an oligofructose overload model in 12 non-pregnant heifers. Clinical parameters and blood samples were obtained at 48 and 24 h and at 6, 12, 24, 36 and 48 h after induction of lameness. Clinical parameters included heart rate, respiratory rate, ruminal frequency and lameness score. Plasma biomarkers included cortisol, haptoglobin, norepinephrine, beta-endorphin and substance P. Differences were observed in all parameters between control and treated heifers. The plasma concentration of biomarkers increased significantly in treated animals starting 6 h after induction of lameness, reaching maximum levels at 24 h for cortisol, 48 h for haptoglobin, 6 h for norepinephrine, 12 h for substance P and at 24 h for beta-endorphin. Overall, our results confirm that lameness associated pain induced using the oligofructose model induced changes in clinical parameters and plasma biomarkers of pain and stress in dairy heifers.