海带幼苗低温度夏技术的回顾与展望

海带(Saccharina japonica)幼苗低温度夏技术,于上世纪50年代建立,至今仍然是我国海带育苗企业普遍采用的育苗方法。尽管该技术在保障幼苗稳定供给,延长海带生长时间（从而提高产量）方面有突出优点,但与其配套的制冷海水循环系统却存在不同海带品种精子相互污染从而导致品种混杂的问题。简要回顾了我国海带幼苗低温度夏技术的发展历史,重点分析了精子污染问题的成因及其对品种维持的负面影响,并对将来的发展趋势进行了展望,以期为我国今后海带的苗种培育和良种品种维护工作提供多元化思路。     

Natural antibodies related to metabolic and mammary health in dairy cows

Natural antibodies (NAb) are defined as antibodies that circulate in normal healthy individuals under the absence of deliberate antigenic stimulation. Two types of NAb are distinguished: NAb towards exogenous antigens and NAb towards autoantigens (N(A)Ab). The objectives of the current study were threefold. First, we studied the relation between metabolic health and concentrations of NAb binding keyhole limpet hemocyanin (KLH) or lipopolysaccharide (LPS) in milk and plasma of dairy cows in early lactation. Second, we determined the presence of N(A)Ab binding transferrin, myosin and thyroglobulin in bovine milk. Third, we studied the relation between N(A)Ab in bovine milk and mammary health. For the first objective, dairy cows were either fed a control (C) (n = 8) or a diet where 2 kg of concentrates were replaced by an iso-energetic concentrate containing marine algae (ALG) from week −3 till 8 postpartum (experiment 1). Plasma and milk samples were analyzed weekly for NAb binding either KLH or LPS. Plasma was analyzed for glucose, non-esterified fatty acids (NEFA) and β-hydroxybutyrate (BHBA). For the second and third objective, milk samples were collected weekly from 96 dairy cows from week 2 till 9 postpartum and analyzed for milk composition and N(A)Ab binding myosin, transferrin and thyroglobulin (experiment 2). For both datasets, N(A)Ab titers are expressed as ²log values of the highest dilution giving a positive reaction. Data are expressed as means ± SEM. Repeated observations were analyzed in a mixed model. In experiment 1, no diet effect (P > 0.05) was observed on NAb binding LPS in plasma or milk, NAb binding KLH in milk was greater (P = 0.05) for cows fed the control diet. Concentration of NAb binding KLH and LPS in plasma was negatively related to plasma NEFA concentration (P < 0.05). In experiment 2, NAb binding myosin (5.66 ± 0.06), thyroglobulin (4.85 ± 0.06), and transferrin (5.76 ± 0.07) were identified in milk. Clinical mastitis incidence (9%) tended to be positively related to concentration of NAb binding myosin (P = 0.06) and negatively related to Nab binding transferrin (P = 0.08). In conclusion, NAb binding KLH and LPS in plasma and milk are related to metabolic health, as indicated by plasma NEFA concentration. Furthermore, this study demonstrates the presence of N(A)Ab in bovine milk and shows trends for a relation between N(A)Ab binding auto-antigens and mastitis. Future studies should confirm these trends and shed light on the predictive value of N(A)Ab in bovine milk for mammary health.     

小剂量肝素对危重新生儿伴全身炎症反应综合征的早期干预作用

目的观察小剂量肝素对危重新生儿全身炎症反应综合征(SIRS)的治疗效果及预后的影响。方法对我科2005年6月至2007年6月间符合诊断标准的94例新生儿SIRS患儿随机分组进行前瞻性对照研究,对照组46例采用常规治疗,治疗组48例在常规治疗基础上每次加用小剂量肝素(5u/kg)皮下注射,每6h 1次,连用3d,观察两组C-反应蛋白(CRP)、血小板计数(PLT)、D-二聚体(D-D)治疗前后的变化。结果治疗组SIRS持续时间>3d者16例(33.3%),明显少于对照组的28例(60.9%),MODS发生率和病死率也明显低于对照组(10.4%、4.2%vs34.8%、17.4%)(P<0.01或0.05);两组治疗后的PLT、CRP及D-D水平均明显改善,且以治疗组更为明显(P<0.01)。治疗组使用肝素后凝血指标均恢复正常或趋于正常。结论早期应用小剂量肝素可有效缩短SIRS持续时间,MODS的发生率及病死率明显降低。     

不同光质处理对库拉索芦荟有效成分的影响

[目的]研究不同光质对库拉索芦荟有效成分的影响。[方法]以库拉索芦荟为试验材料,运用植物化学法和高效液相色谱法测量了红、蓝不同光质处理下,芦荟中可溶性糖、可溶性蛋白和蒽醌类物质的含量。[结果]结果表明,与对照（白光照射）相比,红光照射下可溶性糖、可溶性蛋白和蒽醌类物质的积累有所下降,分别减少了17％、24％、2％;蓝光照射下可溶性糖和可溶性蛋白的含量分别减少33％、27％,而蒽醌类物质的含量增加了44％。[结论]蓝光有利于蒽醌类次生代谢物的积累。     

杜洛克猪精子体外4℃保存过程中活力参数及蛋白修饰变化

为探究体外4℃保存对猪精子活力参数及蛋白修饰的影响,使用猪精液保存稀释液体外4℃保存猪精子7d,利用计算机辅助精子活力分析仪(CASA)分析测定新鲜精液和保存7d后精液的精子活力指标;使用相应试剂盒检测精子中ATP水平及甘油醛-3-磷酸脱氢酶(GAPDH)活性;应用蛋白免疫印迹(WB)技术,比较新鲜精液在4℃保存7d后蛋白翻译后修饰水平的变化。结果表明,猪精液体外4℃保存7d后,精子活力参数呈现下降趋势。其中,运动力(MOT)、快速前进性运动速率(PRO)、平均路径速度(VAP)、平均直线运动速率(VSL)与新鲜精液相比分别降低59.99%、37.03%、12.34%、18.97%,且差异显著(P0.05);发现精子中ATP水平和GAPDH酶活性在4℃保存7d后显著降低(P0.05),即随着体外保存时间的延长,精子细胞内的能量供应不足,代谢水平降低;猪精液4℃保存7d后精子蛋白的磷酸化和酰化修饰明显减弱,这一结果与精子活力、代谢酶活性结果的变化趋势一致。研究结果证实,在体外4℃保存条件下,可能因精子细胞内能量代谢受阻而影响了精子活力及蛋白修饰水平。因此,本研究无论对于丰富精子的能量代谢基础理论,还是对于延长猪精液低温保存时间,提高人工授精效率,均具有重要的理论和实践意义。     

丁香酚麻醉半滑舌鳎成鱼的试验研究

研究了在20～21℃水温下丁香酚对半滑舌鳎成鱼的麻醉作用。根据鱼体在最终麻醉状态和复苏过程中的行为特征,把麻醉程度分为6个时期,复苏过程分为4个时期。鱼体最终麻醉程度不超过3期时,呼吸频率略微增加,到4期时呼吸频率才明显下降。丁香酚麻醉半滑舌鳎成鱼的有效浓度为40～80mg/L,在此范围内,丁香酚浓度越高鱼体达到4期的麻醉时间越短,但复苏时间没有明显差别。丁香酚浓度在40～60mg/L之间时,最长麻醉保活时间>20min;当丁香酚浓度≥80mg/L时,最长麻醉保活时间<8min,鱼体入麻时间与死亡时间的边界很窄,麻醉后应立即进行复苏,否则会导致鱼体死亡。     

兽用生物制品活菌计数方法改进的研究

活菌计数是细菌类活疫苗质量标准中重要的检验参数,与疫苗的保护效果显著相关。现行兽用细菌类生物制品检验中的活菌计数方法部分步骤易产生误差,试验成立标准表述笼统,最终影响到细菌活疫苗质量评判。因此,从检验实际需求出发,参照国内外活菌计数相关规程,通过比对试验研究,对“活菌计数法”中平板干湿程度、菌液滴加方法、菌液分散方法进行改进,提出及验证以标准差(s)衡量结果的有效性并提供参考值。改进后的方法要求更明确,普适性更高,从而减少了操作误差,提升了“活菌计数法”的规范性、可操作性及检验结果可靠性。     

Spermiation time affect the milt quality indices of the Russian sturgeon,Acipenser gueldenstaedtii,Brandt & Ratzeburg, 1833

In this study, the effects of spermiation time are investigated on milt quality of Russian sturgeon over the course of the spawning season. The milt samples were collected from three broodstock batches at three time points including: the beginning, middle and at the end of the spawning season. According to the results, the milt quality parameters including pH, sperm density, spermatocrit, duration of sperm motility and percentage of sperm motility were significantly low in the beginning and end of season than middle of season. The values of milt quality parameters in the middle of season were as follows: (motility percentage: 69.6 ± 3.5, motility duration: 460.3 ± 37.2 s, sperm density: 8.7 ± 0.4 × 10⁹, milt volume: 86.3 ± 8.1 and milt pH: 8.3 ± 0.15). Significant positive correlations were also found between milt pH and sperm motility as well as between sperm density and spermatocrit. In conclusion, our study showed that the middle of season is the best time for collection of milt with appropriate quality in Russian sturgeon. Selection of milt with good quality is necessary aim to cryopreservation of spermatozoa in endangered fish species including Russian sturgeon.     

EM微生物制剂对油菜产量和品质的影响

以油菜为研究对象,采用盆栽试验,研究了不同浓度的EM微生物制剂对油菜产量及品质的影响,EM微生物制剂稀释倍数分别为200、400、600、800,以清水作为对照。结果表明,EM微生物制剂能够增加油菜产量,提高叶绿素、维生素C、氨基酸以及速效磷的含量,并且能够降低有机酸和硝态氮含量,其中以稀释400倍的EM微生物制剂最有利于油菜产量和品质的提高,且EM微生物制剂在一定范围内存在浓度效应。     

斑鳜精液超低温冷冻保存及其效果分析

实验所用翘嘴鳜(♀, Siniperca chuatsi) 2~3龄, 体质量1 000~1 500 g; 斑鳜(♂, Siniperca scherzeri Steindachner)1~2龄, 体质量300~500 g。于繁殖季节选取成熟度好的亲鱼以促黄体素释放激素类似物(LHRH-A2)、绒毛膜促性腺激素(HCG)和马来酸地欧酮(DOM)进行催产。通过筛选D-15、D-17、Ringer液 和 M-Hank’s液4种稀释液和二甲基亚砜 (DMSO)、 甘油(Gly)、乙二醇 (EG)、1,2-丙二醇 (PG)和二甲基甲酰胺 (DMF)5种抗冻剂, 发现DMSO和D-17分别是优选的抗冻剂和稀释液。以D-17作为斑鳜精子的稀释液, 按照1︰3(精液︰稀释液)体积比稀释, 添加10%体积的DMSO作为抗冻剂, 按照三步冷冻法超低温冷冻保存, 37℃水浴解冻的斑鳜精子, 经CASA分析精子活率为(83.26±18.20)%, SCGE检测表明70%以上的精子核DNA不会发生损伤; FCM分析表明26.74%的解冻精子有完整的细胞膜且线粒体功能正常; 用冷冻复苏斑鳜精子与翘嘴鳜精卵进行人工授精, 最高受精率(39.6±6.5)%, 温度24~28℃孵化时间38 h, 孵化后的鱼苗发育正常, 开口1周以后的鱼苗全长(1.346±0.255 ) cm, 体高(0.438±0.103) cm, 体质量(0.045±0.020) g。鲜精受精鱼苗和冻精受精鱼苗在开口后1周的生长没有显著差异。因此认为D-17+10% DMSO可用于斑鳜精液的超低温冷冻保存。本研究将有助于斑鳜种质资源的收集保存和冷冻保存的斑鳜精液在翘嘴鳜(♀)×斑鳜(♂)杂交中的应用。