Prolonging hypothermic storage (4 C) of bovine embryos with fish antifreeze protein

Embryos obtained via superovulation are necessary for mammalian artificial reproduction, and viability is a key determinant of success. Nonfreezing storage at 4 C is possible, but currently used storage solutions can maintain embryo viability for only 24–48 h. Here we found that 10 mg/ml antifreeze protein (AFP) dissolved in culture medium 199 with 20% (v/v) fetal bovine serum and 25 mM HEPES could keep bovine embryos alive for 10 days at 4 C. We used a recombinant AFP isolated from the notched-fin eelpout (Zoarces elongatus Kner). Photomicroscopy indicated that the AFP–embryo interaction was enhanced at 37 C. Embryos pre-warmed with the AFP solution at 37 C for 60 min maintained high viability, whereas those that were not pre-warmed could live no longer than 7 days. Thus, short-term storage of bovine embryos was achieved by a combination of AFP-containing medium and controlled pre-warming.     

咖啡酸苯乙酯通过AMPK/FOXO3a信号通路缓解猪精液常温保存氧化应激的作用机制

【目的】探究咖啡酸苯乙酯（caffeic acid phenethyl ester,CAPE）通过AMPK/FOXO3a信号通路缓解氧化应激对猪精液的影响及其分子机制。【方法】选取8头成年（1—2岁）、健康状况良好长白种公猪进行鲜精采集,将质检合格的样品混池待用。试验设计如下：首先,在常温基础稀释液中分别添加0、0.02、0.04、0.06、0.08和0.10 g·L^-1浓度的CAPE,将精液样品与各个浓度组依次混合后放置室温平衡,随即转入17 ℃电子恒温冰箱进行常温保存。全自动精子分析仪（CASA）测定1—5 d精子运动学参数（总活率与渐进性活力）,筛选出最佳适宜浓度为0.06 g·L^-1。其次,将0.06 g·L^-1 CAPE与400 μmol·L^-1的H₂O₂建立氧化胁迫模型,在第1、3、5天分别采用CASA测定精子总活率与渐进性活力,荧光探针技术评估质膜完整性与顶体完整性,抗氧化试剂盒检测过氧化氢酶（catalase,CAT）与超氧化物歧化酶（superoxide dismutase,SOD）活性,在第5 天利用实时荧光定量 PCR技术测定AMPK/FOXO3a信号通路下游靶向基因（AMPK、FOXO3a、SOD1、SOD2、CAT）及凋亡基因BAX的mRNA表达水平,蛋白免疫印记技术测定AMPK、p-AMPK蛋白表达。【结果】（1）浓度筛选试验中,0.06 g·L^-1的CAPE在保存第3 天显著提高精子的总活率并维持至第5天（P<0.05）,不仅如此,精子的渐进性活力在第1 天显著高于其他处理组（P<0.05）。（2）氧化胁迫试验中,H₂O₂处理组的精子质膜完整性、顶体完整性、总活率、渐进性活力、SOD与CAT活性在保存的第5天均极限显著低于空白组与CAPE+H₂O₂混合组（P<0.001）,而空白组与CAPE+H₂O₂混合组的总活率、顶体完整性、CAT与SOD活性不存在显著差异（P>0.05）。与CAPE+H₂O₂混合组相比,H₂O₂处理组AMPK、FOXO3a、SOD1、SOD2、CAT的mRNA表达水平显著降低（P<0.05）,BAX的表达水平显著升高（P<0.05）,而空白组与CAPE+H₂O₂混合组的AMPK、SOD1、CAT与BAX不存在显著差异（P>0.05）。在蛋白表达水平中,CAPE+H₂O₂混合组与H₂O₂组相比,AMPK、p-AMPK 与p-AMPK/AMPK比率显著提升（P <0.05）。【结论】在常温基础稀释液中添加0.06 g·L^-1CAPE可通过促进磷酸化AMPK的表达,诱导AMPK/FOXO3a信号下游抗氧化分子的转录,继而缓解H₂O₂介导的氧化危害对精液品质产生的影响,延长精液的保存时间,但CAPE保护精子氧化损伤更为深入的分子机制仍需作进一步探究。     

丽江老君山国家公园植物景观资源的开发与保护

党的十八届三中全会提出划定生态保护红线的战略决策,建立国家公园体制是一项与生态红线相关的重要任务。建立国家公园体制的根本出发点是对资源的有效保护与合理利用,其中植物景观资源的开发与保护占据着十分重要的地位。据此,以丽江老君山国家公园为例,就寒温性针叶林、寒温性杜鹃矮林、落叶阔叶林、亚高山草甸4个最能反映该地区植物景观资源特色的植被类型,提出生态观光旅游、科考旅游、休闲旅游以及就地保护和保育景观资源,实施植物恢复工程等开发与保护策略。     

An alkaline phosphatase-based method for the detection of infectious salmon anaemia virus (ISAV) in tissue culture and tissue imprints

Currently, the presence of infectious salmon anaemia virus (ISAV) is often detected in Atlantic salmon by the use of an indirect fluorescent antibody test. This test is limited by the poor stability of fluorescein isothiocyanate which fades after about a week in storage, preventing the development of stained archive material as a reference source. One possible alternative would be the use of immunohistochemical staining methods to detect ISAV. An immunohistochemical method is presented that uses alkaline phosphatase‐conjugated antibodies and Vector^® Red as a substrate, to detect ISAV in kidney imprints. This paper also describes a procedure where Bouin's fluid is used to successfully inhibit endogenous alkaline phosphatase in tissue samples, prior to immunohistochemical processing. This method provides a stable stain that can be read for many weeks after staining or archived for future reference.     

Influence of storage conditions on viability of quiescent copepod eggs (Acartia tonsa Dana): effects of temperature, salinity and anoxia

Copepods have proven to be an ideal source of live food for the production of marine fish larvae in aquaculture. Therefore, there is a need to develop new methods for production and storage of copepod eggs that can be hatched and used at fish farms. In the present study quiescent eggs of Acartia tonsa were stored for periods up to 35 weeks at different temperatures, salinities and oxygen conditions in a full factorial experiment. None of these storage conditions seemed to induce diapause in eggs even though this has been reported by other authors. The most promising storage conditions were those involving low temperature (<5°C), medium salinity (10–20 ppt) and anoxia. The practical aspects of these results for aquaculture are discussed.     

壳聚糖在水果贮藏保鲜中的应用研究

本文首先介绍了壳聚糖的理化性质及生理功能,壳聚糖具有很好的气调性、保湿性和抑菌性,这些性质使得壳聚糖适宜作为水果保鲜剂,目前已被广泛应用在食品行业;然后讨论了壳聚糖的保鲜机理,在此基础上,进一步对壳聚糖膜保鲜水果的应用进行了分类介绍。     

样本预处理方式对罗氏沼虾碳和氮稳定同位素比值检测的影响

为研究样本预处理方式对罗氏沼虾碳和氮稳定同位素比值检测的影响,分别选用3种保存温度(常温23℃/15 d、冷藏2℃/90 d、冷冻-18℃/360 d)、2种取样方式(虾仁取样、整虾取样),以新鲜沼虾样本为对照组,对比不同样本预处理方式对罗氏沼虾碳稳定同位素比值(δ¹³C)、氮稳定同位素比值(δ¹⁵N)、碳氮比(C/N)的影响。结果表明,不同保存温度和不同取样方式对罗氏沼虾δ¹³C、δ¹⁵N、C/N的影响明显。与对照组相比,常温保存条件下的δ¹⁵N偏移量最大(1.31‰~13.16‰,平均值为6.02‰),冷冻保存条件下的δ¹³C偏移量最大(2.34‰~2.84‰,平均值为2.57‰),室温保存条件下的C/N显著增加(P<0.05),冷藏和冷冻保存条件下的C/N显著减少(P<0.05)。不同取样方式对罗氏沼虾δ¹⁵N、δ¹³C的影响还取决于保存温度,在冷冻保存条件下,整虾取样的δ¹⁵N、δ¹³C显著高于虾仁取样,而在常温保存条件下,整虾取样的δ¹⁵N偏移量显著低于虾仁取样。因此,常温(23℃)和冷藏(2℃)条件不适合保存罗氏沼虾、冷冻(-18℃)保存罗氏沼虾的时间不宜超过60 d,建议新鲜沼虾采集后应立即干燥并开展碳和氮稳定同位素检测。本研究结果为水产品碳和氮稳定同位素比值检测的样本预处理方式提供了参考。     

玻璃温室与塑料温室春季保温性能比较

以双屋脊玻璃温室和双脊窗圆拱型薄膜温室为研究对象,观测春季时典型晴天和典型阴天天气条件下2种温室内的气温及地温日变化。结果表明:晴天及阴天时,玻璃温室的日均气温和日均地温均比塑料温室高。晴天时玻璃温室的室内最高气温、最低气温以及日均气温分别比塑料温室高1.7、0.6、1.5℃;玻璃温室的室内最高地温、最低地温以及日均地温分别比塑料温室高1.3、0.9、1.1℃。阴天时玻璃温室的日均气温、日均地温分别比塑料温室高0.5、0.6℃。因此,在保温性能方面,玻璃温室优于塑料温室。     

9个鲜食蚕豆品种低温保鲜比较试验

以苏蚕2号、海门大青皮、上虞田鸡青、美国蚕豆、陵西一寸、通鲜6号、日本大白皮、慈溪大白蚕以及自主研制新品系（长荚型蚕豆）共9个鲜食蚕豆品种为试材,进行低温保鲜比较试验。结果表明：以豆荚形式贮藏,以上虞田鸡青的保鲜效果最好,慈溪大白蚕、日本大白皮和长荚型蚕豆等3个品种次之,以2℃贮藏28d为宜。以豆粒形式贮藏,日本大白皮和慈溪大白蚕等2个品种保鲜效果最好,以2℃贮藏21d为宜。     

臭氧对‘鸭梨’等3种梨果保鲜效果的影响

以‘鸭梨’、‘雪花梨’和‘黄冠梨’为试材,研究了不同浓度的臭氧对梨果品质的影响.结果表明,臭氧处理使‘鸭梨’和‘雪花梨’贮藏期变短,臭氧浓度越大,‘鸭梨’硬度、可溶性固形物、可滴定酸含量下降得越快,果面软化现象发生得越早;臭氧处理延缓了‘雪花梨’可溶性固形物、可滴定酸含量的下降,但使其果点变黑,浓度越大,发生时间越早.臭氧处理延缓了‘黄冠梨’硬度、可溶性固形物及可滴定酸含量的下降,降低了鸡爪病、黑心病的发病率及烂果率,60 mg/m3臭氧处理的‘黄冠梨’贮藏保鲜效果最佳.