斜带石斑鱼肝细胞分离及原代培养方法的建立

本研究以斜带石斑鱼肝细胞为实验对象,在不同培养条件下进行原代培养,旨在探讨稳定可靠的斜带石斑鱼肝细胞分离及原代培养方法。采用组织块分离法和胰蛋白酶(含EDTA)消化法分离肝细胞,并通过密度梯度离心法分离纯化肝细胞,细胞悬液于DMEM/F-12、M199和L-15培养液中培养;细胞活力及数量采用血球计数板计数,并通过MTT法测定细胞增殖率;同时,测定不同时间培养上清液中乳酸脱氢酶(LDH)活性、白蛋白(ALB)和尿素氮(BUN)的含量,以分析肝细胞生长状态。结果表明,组织块方法不适于斜带石斑鱼肝细胞的培养,未见细胞从组织块中迁出,而胰蛋白酶消化法获得良好稳定的培养效果,细胞产量达到1.6×108个/g肝重,活细胞数达到95%;L-15培养基细胞生长明显优于DMEM/F-12和M199培养基;启动原代培养的48~72 h阶段肝细胞生长代谢旺盛,培养上清液中LDH活性显著降低,ALB和BUN含量显著升高。结果显示,0.25%的胰蛋白酶常温消化法适合斜带石斑鱼肝细胞的分离,斜带石斑鱼肝细胞原代培养的最适培养基为L-15培养基,肝细胞在启动原代培养的48~72 h生长代谢旺盛。     

南海区驯养条石鲷亲鱼的初次性成熟和产卵

2008年4月初在广东省饶平县对2006年5月人工孵化和培育的2龄条石鲷（ Oplegnathus fasciatus）成熟亲鱼进行激素诱导,研究和观察条石鲷亲鱼初次性成熟的繁殖生物学。结果显示,在南海区全人工养殖的条石鲷亲鱼初次性成熟年龄为2龄,成熟亲鱼的最小型为全长245 mm、体质量610 g,最大个体为全长300 mm、体质量1450 g;产卵季节为4月10日~7月15日,产卵盛期为4月中旬至6月下旬;为升温产卵型鱼类,产卵温度为20.0~28.8℃,适宜的产卵温度为20.7~27.6℃;雌、雄亲鱼发育同步,个体大小和成熟年龄差别不大;雌鱼属于一年一次分批产卵类型,产卵期超过3个月;24尾初次性成熟雌性亲鱼的总产卵量为3180.0×104粒,日最高产卵量为341.5×104粒;其产卵量和受精卵质量与水温的变化关系密切,受精卵的浮卵率随温度的变化而变化,在水温相对衡定时,浮卵率相对稳定并维持在80%~95%的较高水平;受精卵平均卵径为（0.860±0.023）mm,油球径为（0.191±0.009）mm。     

Soybean molasses as an organic carbon source in the farming of Litopenaeus vannamei (Boone, 1931) in a biofloc system

Soybean molasses was evaluated as a partial replacement for sugarcane molasses as a carbon source for biofloc development in the superintensive culture of Pacific white shrimp (Litopenaeus vannamei). A 50‐day study was conducted with juvenile (3.2 g) shrimp stocked in 16 800 L tanks at a stocking density of 250 shrimp m^?3. Control of total ammonia concentration was performed by the addition of combined mixtures of soybean and sugarcane molasses to the culture water. Three different molasses treatments were evaluated using different soybean‐to‐sugarcane molasses ratios: 15–85%, 38–62% and 60–40% respectively. The control group was treated only with sugarcane molasses. Water quality, chlorophyll a concentration, heterotrophic bacterial load, Vibrio spp. concentration and zootechnical indexes were all evaluated. Total ammonia concentration was controlled by heterotrophic and chemotrophic pathways. Biofloc formation, as quantified by measuring the total suspended solids, was not altered. The Vibrio spp. concentration showed a significant reduction in treatments with soybean‐to‐sugarcane molasses ratios of 38–62% and 60–40%. All combined mixtures of soybean and sugarcane molasses could maintain water quality and productivity in the superintensive culture of L. vannamei using the biofloc system. Thus, the potential use of a residue from agroindustry as a carbon source in a biofloc culture is demonstrated.     

Relationship Between Dietary Protein Source and Feeding Frequency During Feeding Nile Tilapia,Oreochromis niloticus (L.) Cultured in Concrete Tanks

ABSTRACT

A 24-week feeding trial was conducted in concrete tanks with Nile tilapia, Oreochromis niloticus (L.) with an average initial weight and length of 50.87 ± 6.03 g and 14.4 ± 0.45 cm, respectively, to examine the effect of two protein sources and two feeding frequencies on growth performance, production traits, and body composition. Twelve 4 m³ concrete tanks (2 × 2 × 1.25 m, long, width, and height) were each stocked with 100 fish and fed diets containing either fish meal protein or soybean meal protein at two feeding frequency of two times daily or four times daily (2 × 2 × 3 factorial experiment). The results revealed that there was no significant effect of dietary protein sources on growth rate, whereas there was a significant increase in growth rate with increasing feeding frequency (P < 0.05). The same trend was also observed for mean body weight (g), production rate (kg/m3), specific growth rate (SGR % day^?1). The best final mean body weight (g), specific growth rate (SGR % day^?1), and production rate (kg/m3) were recorded in groups of fish fed with feeding frequency four times daily. Whole fish moisture, protein, fat, and ash contents were significantly influenced (P ≤ 0.05) by protein sources and feeding frequency, except protein not influenced with feeding frequency. Energy was significantly (P < 0.05) influenced by feeding frequency, but not by dietary protein sources. Economic evaluation indicated that soybean meal (Diet B) at four times feeding daily was the most cost-effective and affordable feed strategy for farmers. We conclude that a soybean diet fed four times daily is recommended for adult Nile tilapia reared in concrete tanks.     

NaCl胁迫对柱花草幼苗生物学指标的影响

采用盆栽法研究不同NaC(l盐)浓度胁迫对12个柱花草(Stylosanthes)品种幼苗生长的影响。测定叶片危害情况、植株生长高度、地上生物量、地下生物量等生物学指标,采用灰色局势决策模型法进行分析与综合评价。结果表明,随着盐处理浓度的增加,植株表现的萎蔫程度加强,植株的萎蔫速度加快;受胁迫植株的生长高度与对照相比呈下降趋势,受胁迫植株的生物量(茎叶、根)与对照的差异均达极显著。12个柱花草品种最终耐盐性排序为:有钩>热研5号>西卡>GC1480>GC1463>Mineiro>热研7号>GC1581>GC1517>热研2号>热研10号>热研13号。     

东方百合组织培养浅述

综述了东方百合的组织培养技术,包括外植体来源、影响组织培养的因素和组培苗的生根和移栽。     

社会转型期黑龙江乳业文化传承与发展研究——基于文化经济学视角

我国当前处于社会转型期,经济结构正从增量扩能为主转向调整存量、做优增量并举的深度调整.基于文化经济学相关理论,以黑龙江省肇东市为例,利用经济学模型进行分析,进而提出对策建议.从宏观方面,运用态势分析(SWOT)等方法,阐释文化经济发生的动力与意义需求如何产生,并为文化与经济的交互作用奠定基础,研究社会转型时期如何科学传...     

芍药属组间杂交种皮开裂种子的解剖观察与胚培养

【目的】探究芍药属组间杂交种皮开裂种子的内部状态和胚培养成苗表现,为揭示种皮开裂种子的成苗潜力、提高组间远缘杂交育种效率提供参考。【方法】以24个芍药属组间杂交组合种皮开裂的种子为试验材料,解剖后观察并统计胚和胚乳的发育情况,通过离体培养观察胚的萌发和生长规律,分析不同组合种皮开裂种子发育和成苗差异。【结果】芍药属24个组间杂交组合中均出现了种皮开裂的杂交种子,平均种皮开裂率高达61.08%;1 767粒种皮开裂种子中,有11.09%的胚乳已腐烂;胚乳未腐烂种子中,约90%以上种子的胚乳存在不同程度的损伤或病变,24.18%的种子内部未观察到可见胚。对解剖得到的1 185个组间杂种胚进行离体培养发现,有24.73%的胚不萌发,63.47%的胚生长异常,仅有4.47%的胚能正常生长,得到完整植株。胚培养得到的完整植株具备良好的活力,移栽成活率达到86.79%。24个杂交组合种皮开裂种子通过离体胚培养共得到46株移栽成活的组间杂种苗,平均成苗率为2.60%,接近正常组间杂交种子的播种成苗率（2.75%）。【结论】种皮开裂现象在芍药属组间杂交种子中普遍存在,是组间远缘杂交障碍的重要表现之一,用离体胚培养技术能有效克服这类障碍,提高成苗率。     

福建柏愈伤组织诱导与抗褐化研究

【目的】比较不同抗褐化剂减轻福建柏组织培养过程中褐化现象的效果,以提高福建柏愈伤组织培养的增殖分化能力。【方法】以福建柏鳞叶和茎段为外植体,通过单因素试验,研究0.0～2.5 mg/L NAA和2,4 D及0.0～3.0 mg/L IBA对愈伤组织诱导率与生长状态的影响;以福建柏茎段为外植体,采用单因素和多因素组合试验,研究100～2 000 mg/L聚乙烯吡咯烷酮（PVPP）、100～1 000 mg/L活性炭（AC）和V_C以及不同质量浓度VC与AC组合,对愈伤组织污染率、死亡或褐化率及诱导率的影响;以诱导的愈伤组织为材料,采用单因素试验,研究100～1 000 mg/L PVPP、100～300 mg/L V_C、200～1 000 mg/L AC、30～100 mg/L V_B对愈伤组织增殖生长量、增殖率、褐化率及PPO、POD活性的影响。【结果】① 不同质量浓度NAA、2,4-D和IBA对福建柏鳞叶愈伤组织的诱导率总体高于茎段,各生长素对福建柏鳞叶和茎段愈伤组织的诱导率均表现为NAA＞IBA＞2,4-D,所有处理中以1.0 mg/L NAA对福建柏鳞叶愈伤组织的诱导率最高,为83.33%;以0.5 mg/L NAA对茎段愈伤组织的诱导率最高,为61.11%;愈伤组织生长状态和抗褐化效果优劣依次为NAA、2,4-D、IBA。②仅100 mg/L V_C+200 mg/L AC处理的愈伤组织污染率低于不添加抗褐化剂的对照;死亡或褐化率仅500 mg/L PVPP处理与对照相同,均为0.00%,其余处理均高于对照;诱导率则以不同质量浓度PVPP、1 000 mg/L VC、100 mg/L V_C＋200 mg/L AC处理与对照相当或高于对照,其余处理均低于对照;愈伤组织生长状态总体以PVPP处理表现最优。③仅300 mg/L PVPP、200和400 mg/L AC处理的愈伤组织增殖率高于对照,增殖生长量仅100和200 mg/L PVPP、100 mg/L V_C、200 mg/L AC处理高于对照;所有处理中以200 mg/L AC处理的愈伤组织增殖率和增殖生长量最高,分别为70.00%和10.07 g;各抗褐化剂对福建柏愈伤组织增殖效果的优劣依次为PVPP、AC、V_C、V_B。④不同抗褐化剂处理的愈伤组织褐变率均呈先增加后趋于稳定的变化趋势,但变化幅度存在显著差异;各抗褐化剂的抗褐化效果优劣依次为AC、PVPP、V_C、V_B。⑤不同质量浓度PVPP、V_C、AC处理对福建柏愈伤组织PPO和POD活性的影响具有显著差异,但各处理PPO活性均与对照无显著差异,POD活性以500 mg/L AC处理显著高于其他处理。【结论】在福建柏愈伤组织诱导阶段,愈伤组织的褐化与生长素种类有关,生长素NAA、2,4-D适用于福建柏愈伤组织的诱导,IBA诱导的愈伤组织容易褐化,添加适量抗褐化剂能减少愈伤组织的褐化情况。在愈伤组织增殖培养阶段,适宜质量浓度的PVPP、AC作为抗褐化剂能有效减缓福建柏愈伤组织的褐化进程,保持愈伤组织的正常增殖生长,但高质量浓度的抗褐化剂会抑制愈伤组织的生长。     

图书馆事业与中华文化的思考

图书馆是中华文化的资源库,是建设和谐社会的重要文化阵地。繁荣和发展图书馆事业是中华文化建设的需要。