Bacillus subtilis CK3 used as an aquatic additive probiotics enhanced the immune response of crayfish Procambarus clarkii against newly identified Aeromonas veronii pathogen

To identify the aetiology of the disease outbreak in cultured Procambarus clarkii, a dominant bacteria strain from the diseased crayfish hepatopancreas was isolated. It was identified as Aeromonas veronii based on biochemical identification and 16S rDNA sequencing. The symptoms of artificially infected crayfish were similar to those of naturally infected P. clarkii. The A. veronii was sensitive to norfloxacin, ciprofloxacin, ceftazidime, cefuroxime and ceftriaxone antibiotics, while resistant to penicillin, ampicillin, oxacillin and piperacillin. Virulence genes such as the enterotoxin genes (act, alt and ast) and haemolytic toxin genes (hlyA and aerA) were detected. After A. veronii infection, the connections between some liver tubules disappeared, the vacuoles appeared in the brush border and the mitochondria were enlarged. The antagonistic action of previous identified B. subtilis CK3 against A. veronii was also detected. The supernatant of B. subtilis CK3 exhibits a significant bactericidal effect on A. veronii. After B. subtilis CK3 immersion, the antioxidant enzymes and immune-related enzyme activities in hepatopancreas were significantly higher than control. The accumulative mortality caused by infection of A. veronii can be significantly reduced by adding B. subtilis CK3 into the aquatic water. These results demonstrated that B. subtilis CK3 could act as a water additive to improve the immune response of P. clarkii against newly identified A. veronii. The present study will provide a new way for the prevention and control of crayfish bacterial diseases and also provide technical support for the healthy cultivation of P. clarkii.     

复合益生菌对眼斑双锯鱼(Amphiprion ocellaris)肠道消化酶、菌群结构以及形态的影响

为探讨饲料中添加复合益生菌对眼斑双锯鱼(Amphiprion ocellaris)肠道消化酶、菌群结构及形态的影响,将135尾初始体重(0.79±0.01)g的眼斑双锯鱼随机分为3组,每组3个重复,分别投喂复合益生菌有效活菌添加量为0%(对照组)、3%(L3组)和6%(L6组)的饲料,进行为期28 d的养殖试验。结果显示,复合益生菌能提升试验鱼肠道蛋白酶的活性,其中L3组肠道蛋白酶显著提升(P<0.05);能显著降低试验鱼肠道淀粉酶活性(P<0.05);能降低脂肪酶活性(P>0.05)。随着饲料中益生菌添加量的增加,试验鱼肠道中变形菌门丰度开始增多,拟杆菌门丰度降低。饲料中益生菌添加量的增加显著降低试验鱼肠道肌层厚度并显著增加试验鱼肠道绒毛高度(P<0.05)。可见,饲料中添加3%复合益生菌可改善眼斑双锯鱼肠道消化酶活性,提高肠道微生物群落多样性,改善肠道组织形态和功能。     

长江常熟溆浦段中华鲟幼鱼出现时间与数量变动

2002-2009年,依托长江渔业资源管理委员会办公室设立在长江常熟溆浦江段的渔业资源监测网(定置张网),对降河中华鲟(Acipenser sinensis)幼鱼进行了监测分析.结果表明,中华鲟幼鱼每年主要于5-7月在常熟溆浦江段出现,出现时间平均86.6 d(n=8),所采获样本(n=8)全长、体长和体质量年度平均值的变化范围分别为25.2～40.1cm、23.2～33.5em和70.7～314.8g.各年采获样本数量差异较大,从2002至2009年呈现出较明显的递减趋势,各年采获样本数量最多的旬也不一致,变化时间为5月中旬至6月下旬.从幼鲟生长来看,不同年份相同时期采获样本的规格差异较大,同一年份同一时间采获样本的规格也存在一定的差异.与三峡工程截流以前进行比较,中华鲟幼鱼在长江口出现的时间没有明显变化,但出现较高数量比例的时间由以前的6月变为5月中下旬和6月,幼鲟规格较截流前也有较大变化,推测造成这种改变的原因可能是因为人工增殖放流活动和三峡工程截流所导致的水文状况的变化.分析认为,水文状况可能是影响中华鲟幼鱼到达长江口时间的重要因素之一.     

我国5个青蛤地理群体形态差异分析

运用3种多元统计分析方法,比较研究了我国青蛤5个地理群体(福建、浙江、江苏、天津及辽宁)的形态差异。选择了14个可量性状和6个地区生态特征参数进行主成分分析和聚类分析。结果表明,江苏、天津和辽宁的形态较接近,其中福建群体的趋异差异最大。主成分分析构建了3个主成分。其贡献率:主成分1为28.05%,主成分2为18.6%,主成分3为14.0%,累计贡献率为60.65%。逐步判别选入8个贡献率较大的性状进行判别分析。结果显示,5个群体形态差异显著(P<0.01)。建立了5群体判别函数,其判别准确率P1为35%~90%,P2为73.7%~86%。综合判别率为67.5%。青蛤形态学差异是遗传和环境共同作用的结果,结合分子遗传学研究将更准确地描述青蛤5群体间的遗传特性,为今后种质资源保护及遗传育种提供依据。     

鳜鱼和奥利亚罗非血乳酸脱氢酶酶谱的比较

哺乳类动物各种组织和器官的乳酸脱氢酶(LDH)同工酶较稳定,酶带分布非常有规律,同源组织的LDH同工酶酶谱通常相似和稳定。鱼类LDH同工酶复杂得多,有三分之二品种中,其LDH同工酶类似哺乳类动物的酶谱;但在其余三分之一种类中,两个亚基随机结合受到限制,出现了不规则酶谱,有的只有2—3条酶带,有的超过5条酶带。即使具有5条酶带的鱼中,其骨骼肌和心肌LDH在电泳酶谱上迁移快慢正好与哺乳类动物相反。此外鱼类C基因产物并不特定在某一组织或器官中,一些鱼的     

棚架下垂叶幕与水平叶幕对香百川葡萄叶片光合特性的影响

[目的]比较棚架下垂叶幕和水平叶幕下葡萄叶片臭氧伤害状况以及对光合性能的影响.[方法]以5年生的葡萄品种香百川为试材,利用臭氧监测仪监测7-9月份栽培环境中臭氧浓度,采用CIRAS-3便携式光合作用测定仪和Handy PEA测定2种叶幕叶片的净光合速率和荧光特性.[结果]下垂叶幕缓解了臭氧伤害情况,其主梢叶片发生臭氧伤...     

文蛤卵母细胞卵黄发生的超微结构

取不同发育阶段文蛤 (MeretrixmeretrixLinnaeus)活体解剖取出生殖腺 ,常规方法制成切片 ,以透射电镜(TEM )观察其卵母细胞的卵黄发生过程。结果表明 ,文蛤的卵母细胞卵黄发生期间 ,线粒体、内质网、高尔基复合体、溶酶体、微吞饮泡等细胞器均参与了卵黄粒的形成。卵黄合成早期的卵母细胞质膜伸出大量的微绒毛 ,并出现卵黄膜 ,胞质中有大量膜性小泡 ,溶酶体、线粒体、高尔基复合体和粗面内质网发达 ,卵黄前体物质逐渐增多。在卵黄合成中期 ,胞质内线粒体和内质网活动活跃 ,卵母细胞大量合成和积累卵黄物质 ,细胞质膜外凸 ,与外界进行物质交换。卵黄合成后期 ,卵质内贮存了大量的卵黄粒 ,细胞器不发达。此外 ,还对卵母细胞发育过程卵黄颗粒的细胞内、外原料来源进行了讨论     

The siderophore-interacting protein is involved in iron acquisition and virulence of Riemerella anatipestifer strain CH3

Riemerella anatipestifer causes epizootic infectious disease in poultry and serious economic losses especially to the duck industry. However, little is known regarding the molecular basis of its pathogenesis. The ability to acquire iron under low-iron conditions is related to the virulence of a variety of bacterial pathogens. In this study, a sip (Riean_1281) deletion mutant CH3Δsip was constructed and characterized for iron-limited growth, biofilm formation, and pathogenicity to ducklings. Results showed that siderophore-interacting protein (SIP) was involved in iron utilization and the sip deletion significantly reduced biofilm formation and adherence to and invasion of Vero cells. In addition, the sip gene was absent in 1 of 24 (4.17%) virulent strains and 2 of 3 (66.7%) avirulent strains of R. anatipestifer, and the sip gene from six R. anatipestifer strains, which belong to serotypes 1, 2, and 10, respectively, shared 100% amino acid identities to those of R. anatipestifer strains DSM15868 and RA-GD. These results suggested that siderophore-mediated iron acquisition may be an important iron-uptake pathway in R. anatipestifer. Animal experiments indicated that the median lethal dose of the CH3Δsip mutant in ducklings was about 35-fold higher than that of the wild-type CH3 strain. Thus, our results demonstrated that R. anatipestifer SIP was involved in iron acquisition and necessary for its optimal virulence.     

鸡肉和鸡蛋中金刚烷胺与金刚乙胺残留检测UPLC-MS/MS法研究

建立了鸡肉和鸡蛋中金刚烷胺与金刚乙胺残留检测的超高效液相色谱-串联质谱方法（UPLC-MS／MS）。样品在酸性条件下乙腈提取后,用正己烷去除脂肪,高速离心去除蛋白质等杂质,上机测试。液相色谱条件：色谱柱为BEHC18（50×2．1mm,1．7μm）,流动相为0．1％甲酸乙腈溶液＋0．1％甲酸水溶液,流速0．3mL／min,柱温30cI二,进样量10μL。质谱条件：电喷雾离子源（ESI^＋）,多反应监测（MRM）方式进行采集。结果表明：金刚烷胺与金刚乙胺在2～100ng／mL浓度范围内均呈现良好的线性关系,相关系数（R。）分别为=0．9977和0．9988;鸡蛋和鸡肉中金刚烷胺与金刚乙胺残留检测方法检测限均为1ng／g,定量限均为2ng／g。从鸡肉组织在2、5和10ng／g三个添加浓度检测结果可以看出,金刚烷胺与金刚乙胺的平均回收率分别为79．6％～107．5％和78．4％～101．2％,批内和批间RSD均小于15％。从鸡蛋在2、5和10ng／g三个添加浓度检测结果可以看出,金刚烷胺与金刚乙胺的平均回收率分别为90．8％～111．1％和75．4％～87．4％,批内和批间RSD均小于15％。该方法具有简便快捷、灵敏度高、定性准确等特点。     

复合蛋白酶在即食海参加工中的应用

用复合蛋白酶处理盐渍海参,作为加工冻干即食海参的预处理样品,研究其酶处理最适工艺。试验结果表明,酶处理温度40℃,处理50 m in,水浸泡13 h,酶用量为肉重0.075%时,处理后的海参胶原蛋白溶解度(DD%)为(38.14±0.52)%,与水发海参接近[(39.34±0.47)%],口感达到最佳。从酶处理海参和盐渍海参的红外光谱分析可知,二者的营养成分无明显差异,达到生产冻干即食海参的要求。