单层细胞共培养对猪去卵丘卵母细胞体外成熟和孤雌发育的影响

本试验比较观察第一极体(The first polar body,PbⅠ)、Oosight imaging system观察和hocchst33342染色法对第2次减数分裂中期(Metaphase Ⅱ,MⅡ)卵母细胞判定结果的相关性分析,并探讨卵巢皮质细胞(porcine ovarian cortex cells,pOCCs)、猪输卵管上皮细胞(porcine oviductal epithelial cells,pOECs)和猪卵丘颗粒细胞(porcine cumulus cells,pCCs)等3种单层细胞体外共培养体系对猪去卵丘卵母细胞(cumulus cells denuded oocytes,Dos)体外成熟(in vitro maturation,IVM)和孤雌发育的影响。结果显示:(1)Oosight imaging system判定卵母细胞成熟的结果与hocchst33342染色法判定的结果有很强相关性(R=0.973,P<0.01,N=90);(2)pOCCs单层细胞共培养体系中猪去卵丘卵母细胞成熟率显著高于pOECs((52.5±0.30)%vs(43.8±2.18)%,P<0.05),且...     

循环水系统内马氏珠母贝与3种大型藻类的混养状况及水质分析

为探讨马氏珠母贝与大型海藻的最佳混养体系,分别设置马氏珠母贝稚贝与针叶蕨藻、麒麟菜、长茎葡萄球藻混养实验组及马氏珠母贝稚贝对照组,记录养殖过程中养殖水体NH_4~+-N,NO_3~--N的浓度变化及马氏珠母贝稚贝的存活率。结果表明,实验组在养殖过程中可保证较好的水质状况,稚贝存活率也高于对照组;长茎葡萄球藻混养实验组对养殖水体的NH_4~+-N,NO_3~--N处理效果最优,分别在第47天和第49天达最低值[(0.015±0.003)mg·L~(-1)和(0.266±0.002 mg·L~(-1)],该组的稚贝存活率和藻体日增体质量(0.86 g·d-1)均最高(93.33%);不同混养实验组的马氏珠母贝平均壳高日增长量均显著高于对照组。     

四种抗生素对共培养的萱藻丝状体和膨胀色球藻的影响

为抑制或消除萱藻丝状体扩增阶段出现的膨胀色球藻,本研究探讨了头孢噻肟钠、阿莫西林、四环素和红霉素4种抗生素对共培养的萱藻丝状体和膨胀色球藻产生的影响。结果显示,浓度为50和100 mg/L的头孢噻肟钠均显著抑制膨胀色球藻的生长,进而保证萱藻丝状体正常生长与发育;浓度为50~1 000 mg/L的阿莫西林对膨胀色球藻和萱藻丝状体均无抑制作用,阿莫西林不适于去除共培养体系中的膨胀色球藻;浓度为100和200 mg/L的四环素对膨胀色球藻有显著的抑制作用,但此浓度下萱藻丝状体细胞质萎缩,生长状况较差;浓度为0.10~1.00 mg/L的红霉素对膨胀色球藻的生长均有显著的抑制作用,但当其浓度超过0.50 mg/L时,萱藻丝状体的生长亦受到抑制,甚至死亡。     

影响西洋参不定根组培增殖的几种因素及皂苷生产的研究

为筛选西洋参不定根组培增殖的适宜培养基,研究培养基中生长素IBA和NAA、磷浓度及有机物含量对不定根增殖生长的影响,结果表明,不定根在IBA 5 mg/L中生长良好,MS培养基中固有磷浓度和有机物含量适合于西洋参不定根的生长.为有效生产西洋参皂苷,利用生物反应器进行不定根与细胞的共培养,并与反应器不定根和细胞培养进行比较,发现共培养时获得的不定根生物量（鲜重和干重）与不定根单独培养时无明显差异,共培养的细胞生物量不及细胞单独培养,但共培养的培养物（细胞＋不定根）总生物量大于细胞或不定根单独培养;测定皂苷含量的结果,共培养时反应器中不论是不定根还是细胞,其皂苷含量均高于不定根单独培养的不定根和细胞单独培养的细胞中的含量;皂苷总生产量在共培养时达到26 mg/L,约为不定根和细胞培养的2倍.因此,反应器共培养方法在西洋参皂苷的工业化生产有较高的应用价值.     

两株红树林真菌共培养液抗植物病菌成分研究

从南海红树林内生真菌E33和K38混合发酵液提取物中寻找有抗植物病原菌活性的化合物,利用硅胶柱层析共分离得到6个化合物,用波谱技术分别鉴定后,确定为对苯二甲酸二甲酯、4-甲氧酰基苯甲酸乙二醇酯、4-甲基-2-甲酰基-3,5-二羟基苯甲酸乙酯、rubramin、对羟基乙酰苯乙胺和尿嘧啶。4-甲基-2甲酰基-3,5-=羟基苯甲酸乙酯首次以天然产物的形式被发现,并首次报道了其2D-NMR数据,生测结果表明其对小麦赤霉、大豆疫霉具有中等的抗菌活性。     

稻虾共作模式下稻田pH对土壤和水稻重金属含量的影响

为探究稻虾共作模式对稻田土壤和水稻中重金属含量的影响,以湘晚籼12号、黄华占、玉针香为试验材料,常规稻作模式为对照,在不同土壤背景条件下（试验地点分别位于湖北省荆门市掇刀区和黄冈市浠水县）开展大田试验。结果显示：在弱碱性土壤背景下,稻虾共作模式中稻田土壤中Cr、As、Cd、Pb和Hg的含量降低,水稻根、茎、穗以及糙米中Cd、Pb含量降低;在弱酸性土壤背景下,稻虾共作模式中稻田土壤中Cr、As、Cd、Pb和Hg的含量升高,水稻根中Cd、Pb含量升高,水稻茎叶穗中Cd含量升高,Pb含量降低,水稻糙米中Cd、Pb含量均降低。本研究结果表明,不同pH土壤背景下稻虾模式对土壤和水稻重金属含量的影响表现不一致,但稻虾模式均可明显降低稻米中Cd和Pb的含量。     

稻鱼共生系统的推广潜力分析——以中国南方10省为例

稻田浅水环境为许多水产动物提供了生境,也为稻鱼共生产业的发展提供了基础。但是,一个区域的稻田是否适合发展稻鱼共生系统,常常受到当地自然和社会条件的影响,了解这些影响对有效推广稻鱼共生系统有重要意义。本文以我国南方10省为研究区域,从自然和社会经济因素两个方面分析了稻鱼共生系统的推广潜力。研究利用气象资料和农业统计数据,基于地理信息系统构建了研究区域内的稻田地理分布数据库,确定了15个影响稻鱼共生系统推广效率的指标,通过指标的层级模型和线性加权评分法对不同稻田的推广优先等级进行评估,并构建了基于稻田总面积、推广率和单产水平下鱼产量估算的简易模型。结果表明,综合自然和社会经济条件,研究区域内的稻田可划分为4个推广优先等级,面积占比分别为：等级1占29.6%（3.59×10⁶ hm²）,等级2占16.9%（2.05×10⁶ hm²）,等级3占24.2%（2.94×10⁶ hm²）和等级4占29.4%（3.57×10⁶ hm²）;其中湖南、四川、江西和浙江4省的稻田50%以上属于等级1和等级2,而在云南和贵州基本上所有的稻田都属于等级3和等级4。等级1和等级2的稻田适合推广集约型稻鱼共生模式,这两个等级的稻田每个生长季可产出最高鱼产量分别为3.77×10⁶ t和2.15×10⁶ t;而等级3的稻田适合进行粗放型和集约型模式相结合的推广方式,粗放型和集约型模式最高鱼产量分别为0.62×10⁶ t和3.09×10⁶ t。本研究主要结果为合理制定稻鱼共生系统推广策略提供借鉴,也可为国内外稻田水产养殖产业发展规划的制定提供参考。     

大豆和大豆疫霉菌共培养体系的构建

 本研究旨在建立一个适于分析大豆遭受大豆疫霉菌侵染后基因表达研究的互作体系。在比较了15个携带不同抗病基因的大豆基因型的组织培养情况和7个大豆疫霉菌分离物及其游动孢子单孢系的毒力变化的基础上,构建了大豆悬浮细胞和大豆疫霉菌游动孢子的共培养体系,进而分析了共培养过程中大豆细胞的活力和防御基因表达情况。结果表明,不同亲和力菌株的游动孢子引起的大豆细胞活力变化十分相似;非亲和菌株的游动孢子可诱导寄主防御基因的表达发生变化。此系统为深入开展大豆抗性机制研究提供了良好的平台。     

Bioprocess enhancement of feather degradation using alkaliphilic microbial mixture

1. The main aim of this work is to develop a robust method to generate a microbial mixture which can successfully degrade poultry feathers to overcome environmental problems.

2. Four different alkaliphilic microbes were isolated and shown to degrade poultry feathers.

3. Two of the isolates were phylogenetically identified as Lysinibacillus and the others were identified as Nocardiopsis and Micrococcus.

4. The best microbial co-culture for white and black feather degradation was optimised for pH, temperature and relative population of the isolates to achieve almost 96% of degradation compared with a maximum of 31% when applying each isolate individually.

5. The maximum activity of keratinase was estimated to be 1.5 U/ml after 3 d for white feathers and 0.6 U/ml after 4 d for black feathers in a basal medium containing feather as the main carbon source. Additionally, non-denaturing polyacrylamide gel electrophoresis showed 4 and 3 protease activity bands for white and black feather, respectively.

6. This study provides a robust method to develop potential new mixtures of microorganisms that are able to degrade both white and black feathers by applying a Central Composite Design.     

Granulosa cells in three-dimensional culture: A follicle-like structure for domestic cat vitrified oocytes

Follicle-like structures are three-dimensional matrices joint with living cells that allow the in vitro development of female gametes in more physiological conditions. They have been shown to be beneficial to fresh oocytes in different species, and in this study, domestic cat (Felis silvestris catus) granulosa cells were used to create a functional follicle-like structure aimed at supporting the in vitro maturation of conspecific vitrified oocytes, key players of fertility preservation programmes that usually struggle to acquire their full developmental competence after warming. Cat granulosa cells were cultured for up to 6 days in three-dimensional barium alginate microcapsules (i.e. follicle-like structures) or in two-dimensional monolayers, and their steroidogenic ability (estradiol and progesterone secretion) was assessed to confirm their functionality. The same systems were used (on day 2 or 6 of granulosa cells culture) for the in vitro maturation (IVM) of Cryotop^® vitrified immature cat oocytes and compared with microdrops of IVM medium without cells (control). Granulosa cells were able to maintain their functionality in vitro in both the conditions, even if with a different extent of hormonal secretion along culture (p = .02). Vitrified oocytes resumed meiosis at higher rates when cultured with 2 days old granulosa cells (p = .03), but full maturation rates slightly raised when granulosa cells were cultured longer, albeit without differences with the control group. This study paved the road for the creation of enriched culture systems in the domestic cat, but innovations are strongly needed for vitrified oocytes that deserve better chances to develop in vitro.