非人灵长类动物体外受精与胚胎移植的研究进展

简要介绍了非人灵长类动物体外受精与胚胎移植的内容、方法和步骤,影响其体外受精的主要因素,以及对未来非人灵长类动物的研究方向。     

施肥对复垦土壤微生物代谢功能与苗期玉米生长的影响

利用Biolog-ECO技术、WinRHIZO扫描仪及其图像分析技术,研究了对照CK（施肥量为0）、施复合肥F（施肥量为600 kg·hm^-2）、施有机肥O（施肥量为7 500 kg·hm^-2）、有机无机肥配施OF（施肥量为复合肥300 kg·hm^-2+有机肥3 750 kg·hm^-2）处理下煤矸石复垦区土壤微生物群落代谢功能及苗期玉米根系形态等特征,结果表明：不同施肥处理下煤矸石复垦区土壤微生物群落功能多样性差异显著,OF处理下土壤微生物多样性Shannon-wiener指数与Simpson指数最高,分别为3.22和0.96, Eveness指数最低,为0.21,而CK处理与之相反,Shannon-wiener指数与Simpson指数最低,分别为3.07和0.95,Eveness指数最高,为0.23;影响该区域土壤微生物群落代谢功能多样性的关键碳源包括6种氨基酸（L-精氨酸、L-天冬酰胺酸、L-苯基丙氨酸、L-丝氨酸、L-苏氨酸、甘氨酰-L-谷氨酸)、4种酯（D-半乳糖酸γ内酯、丙酮酸甲酯、吐温40、吐温80）、1种胺（腐胺）、5种酸（D-半乳糖醛酸、2-羟苯甲酸、4-羟基苯甲酸、r-羟基丁酸、D-苹果酸）、3种糖（-甲基D-葡萄糖苷、葡萄糖-1-磷酸盐、a-环状糊精）、3种醇（I-赤藻糖醇、D,L-a-甘油、D-甘露醇）;OF处理下玉米根系形态参数如根长、根体积、根系分形维数及玉米生物性状如生物量、茎粗、株高等与CK处理差异最为显著,说明OF处理有利于煤矸石复垦土壤质量的恢复与提高,利于煤矸石复垦区玉米生长。     

包气带土壤pH对灌溉施肥响应的变异过程

为了分析灌溉施肥活动引起的包气带土壤pH值变异特征及其对地球化学条件的响应,通过历时3 a的野外原位灌溉施肥试验,应用不同季节灌前、灌后6 m土层中不同深度的测定资料,系统分析了土壤pH对灌溉、施肥的响应过程,结果表明：各深度pH值呈弱变异性（CV=1.01%~2.28%）,与灌溉前相比,灌后土壤pH值的均值和变异系数均呈现明显的变化;灌前包气带各层pH具有强烈的空间自相关性,灌后受水分、基质等相互作用影响,pH的空间自相关性有所减弱,C₀/(C₀+C)和变程a分别由7.23 m和3.54 m(灌前0 d)减少到3.26 m和2.76 m(灌后第10天)。土壤基质是决定土壤酸碱性的主要因素,在灌溉施肥活动对pH的响应过程中,地球化学条件（土壤含水量、土壤温度、土壤有机质(SOM)、氧化还原电位（RP）等)、土壤基质组成和氮底物浓度（NH⁺₄-N）等的交互作用影响pH的动态。土壤含水量和温度单独对pH影响不显著,两者交互作用对pH有显著影响。引起土壤pH变化的主要变异源为Cl^-、土壤有机质(SOM)、NO^-₃-N、NH⁺₄-N等营养物质和不同空间深度土壤基质的差异,表明灌溉施肥改变了包气带pH地球化学动力场、营养物质和土壤基质的交互作用,引起各深度的生物地球化学反应,控制pH值的空间变异特性。当包气带介质土壤水分变化时,首先营养物氨态氮以分子态或水合态形式被介质吸附,H⁺得到释放,使得灌后第4天pH值下降。随着氨氧化过程中H⁺的释放,pH在灌前和灌后第10天和第30天有显著差异。氨的氧化引起硝酸盐含量不断增加,使得硝酸盐对pH值的影响在灌后不断增强,相关系数由0.24(0 d,P<0.05)增加到0.41(30 d,P<0.01),而氨态氮对pH值的影响逐步降低,相关系数由0.43(0 d,P<0.01)降低为0.19(30 d,P>0.05)。     

不同施肥模式对早稻季农田氮磷径流流失的影响

为探究湖南双季稻区早稻季防控稻田氮、磷养分流失污染的施肥模式,通过田间试验,设置了不施氮磷肥处理(CK)和常规施肥(CF)、有机肥替代(OM)、控释肥减施(CRF)、绿肥还田(GM)4种施肥模式,研究了不同施肥模式对稻田氮、磷养分径流流失的影响。结果表明:相较于常规施肥模式,有机肥替代、绿肥还田和控释肥减施模式稻田总氮径流流失量分别减少了12.80%、16.62%、28.55%,各施肥处理早稻总氮素流失率大小表现为:常规施肥有机肥替代绿肥还田控释肥减施,氮素流失形态主要以可溶性氮为主,占流失总氮的80.48%~91.96%,可溶性氮中以铵态氮为主。控释肥减施和绿肥还田模式均能减少稻田磷素径流损失量,与常规施肥模式相比,总磷径流流失量分别减少了6.26%和28.30%;有机肥替代模式稻田总磷径流损失量较常规施肥模式增加26.33%;各施肥处理早稻总磷流失率表现为:有机肥替代常规施肥绿肥还田控释肥减施,磷素流失形态前期以颗粒态磷流失为主,后期以可溶态磷为主。在4种施肥模式中,控释肥减施和绿肥还田模式能降低稻田氮磷径流流失量,在南方双季稻区推广这两种施肥模式可有效防控农田氮、磷流失污染风险。     

稳定同位素技术分析不同养殖方式下鳙饵料的贡献率

应用碳氮稳定同位素技术分析了不同养殖方式(A组:施肥;B组:施肥+1/2投饲;C组:施肥+投饲;D组:投饲)围隔中鳙(Aristichthys nobilis)肌肉碳氮稳定同位素变化情况,及其可能摄食饵料的贡献率。结果表明,A组鳙肌肉的δ13C和δ15N值分别为(–23.8±0.1)‰和(10.8±0.4)‰,显著高于B组、C组和D组的值(P0.05),后三者之间值没有显著差异。围隔中的食物源有浮游动物、颗粒有机物和饲料,它们的δ13C和δ15N值均低于消费者鳙的同位素值。3种食物在不同养殖方式下的贡献率不同,浮游动物是A组鳙的主要食物来源,平均贡献率为(65.6±3.2)%,饲料是另3个组鳙的主要食物来源,其中B组饲料的贡献率相对较大,达82.1%。说明投饲鳙后,鳙对饲料能够较好地吸收利用,但在池塘中同时培育天然饵料能有效提高饲料的贡献率,减少投饲量,降低残饵的污染。本研究旨为实践养殖中合理投饲鳙的管理技术提供理论依据。     

Fertilization rate,motility, lipid peroxidation and pH changes after chilled storage of rainbow trout (Oncorhynchus mykiss) eggs and spermatozoa by a RMPI medium

The aim of this study was to evaluate the effects of Roswell Park Memorial Institute (RPMI) 1,640 medium on chilled storage of eggs and spermatozoa of rainbow trout (Oncorhynchus mykiss). After 3 days of storage, eggs in RPMI 1,640 media (pH 8.2, 9 and 10), Cortland medium and coelomic fluid were inseminated with fresh spermatozoa (Experiment 1). Eggs in RPMI 1,640 medium at pH 8.2 shown the lowest thiobarbituric acid‐reactive substances (TBARS, 0.053 ± 0.003 nmol/ml) and pH changes (from 8.20 ± 0.01 to 8.18 ± 0.01), the highest fertilization rate (82 ± 3%). Undiluted and diluted spermatozoa at ratios of 1:2 and 1:9 with RPMI 1,640 media (pH 8.2, 9 and 10) and Cortland medium were inseminated with fresh eggs (Experiment 2). Spermatozoa in RPMI 1,640 medium at pH 9 (1:9) caused the lowest TBARS content (0.037 ± 0.002 nmol/ml) and pH changes (from 9.00 ± 0.01 to 8.98 ± 0.01), the highest fertilization rate (77 ± 2%) and motility parameters. Based on Experiments 1 and 2, eggs and spermatozoa were stored for another 3 days in RPMI 1,640 medium at pH 8.2 and 9 (1:9) respectively (Experiment 3). Fertilization rate of storage eggs and spermatozoa in Experiment 3 was 79 ± 5%, showing successfully storage of rainbow trout gametes with the same medium.     

Production of inbred larvae through self‐fertilization using oocytes and cryopreserved sperm from the same individuals after sex reversal in eastern oyster Crassostrea virginica

The eastern oyster Crassostrea virginica can change sex which makes self‐fertilization possible if sperm can be cryopreserved. In this study, small (~1 year old) and large (~2–3 years old) oysters were biopsied for sperm collection. Survival of the biopsied oysters after 1 year was 50% for small oysters and 17% for large oysters. Oocytes were collected from sex‐reversed females, and self‐fertilized with cryopreserved sperm. Of the 24 cryopreserved samples, 14 individuals had ≤1% fertility when crossed with oocytes from unrelated females, indicating that the cryopreserved sperm had reduced fertility. The other 10 individuals had a fertility of 39 ± 25% when crossed with oocytes from unrelated females (non‐selfing), but showed a significantly lower success of self‐fertilization (12 ± 16%) (P = 0.008), while aliquots of the same oocytes had a fertilization of 83 ± 11% when crossing with fresh sperm. Larvae were produced at day 3 in the self‐fertilized families (12–94% of the fertilized oocytes), and survived to eyed‐larvae stage at days 11–14. Genotyping with 9 microsatellite markers confirmed that the larvae resulted from self‐fertilization in four families. This study demonstrated the feasibility of creating self‐fertilized inbred lines of oysters by use of non‐lethal sperm collection and cryopreservation.     

疣吻沙蚕胚胎发育观察及盐度对其孵化的影响

采用显微镜观察了疣吻沙蚕(Tylorrhynchus heterochaeta)胚胎发育不同时期特征,并依据疣吻沙蚕人工孵化经验设置6个盐度梯度组(A:0,B:5,C:10,D:13,E:15,F:20),每组3个平行,研究了盐度对疣吻沙蚕受精率和孵化率的影响。结果表明,疣吻沙蚕胚胎发育可分为受精卵、卵裂期、囊胚期、原肠期、前担轮幼虫期、后担轮幼虫期、膜内三刚节疣足幼虫期和三刚节疣足幼虫期8个时期;盐度对疣吻沙蚕孵化影响显著(P0.05),C组受精率和孵化率最高,与其他组存在显著差异(P0.05)。适宜受精的盐度范围为10~13,最适孵化的盐度为10。     

贝类受精细胞学的研究现状

从成熟卵子的核相与体外促熟,精子的顶体反应,受精膜的形成,极体的释放与雌雄原核的形成以及多精入卵方面综述了贝类受精细胞学的研究成果,并展望了这一领域的研究和应用前景。     

Towards sustainable exhibits – application of an inorganic fertilization method in coral reef fish larviculture in an aquarium

Coral reef fish are collected from the wild and exhibited in aquaria worldwide. Some of the fish spawn in captivity; however, the eggs are usually neglected. In this study, we collected the eggs spawned naturally in the exhibit tanks, hatched and cultured them indoor in 2000‐L fibreglass tanks (initial density = 18 000 egg tank^?1). We applied an inorganic fertilization method commonly used in freshwater fish culture in raising these coral reef fish larvae. We maintained inorganic phosphorus concentration at 100 μg P L^?1 and inorganic nitrogen at 700 μg N L^?1 daily in the fertilized group (n = 4), while the control tanks (n = 4) were fed with rotifers (10 ind mL^?1). Chlorophyll a at particle sizes of both 0.45–20 μm and >20 μm, as well as NH₃‐N, NO₃‐N, and PO₄‐P concentrations were significantly higher in the fertilized group than the control. Zooplankton in the size groups of 10–50 μm (mainly flagellates) and 50–100 μm (mainly ciliates) were abundant (about 10~60 ind mL^?1) during 3–7 days in fertilized tanks. The average larval fish survival rate at 21 day after hatch in fertilized group was consistently higher than the control in two trials. The experiments demonstrated that the inorganic fertilization approach can be successfully adapted for coral reef fish culture in an aquarium to achieve sustainable exhibits.