不同条件延缓未受精卵激活对鱼类细胞电融合的影响

泥鳅未受精卵先在合成卵巢液、郝氏液中分别浸泡15、20、30min．然后再受精孵化，统计发育情况，结果表明，合成卵巢液可有效延缓未受精卵遇水后被激活的时间，郝氏液对延缓未受精卵被激活也有一定作用，但效果比合成卵巢液差。用合成卵巢液配制不同浓度胰酶处理受体卵进行细胞电融合实验．胰酶质量浓度为5mg／mL时．融合卵发育至囊胚期的比例比对照组（郝氏液）高2％．统计分析表明该差异是显著的（P〈0．05）；但发育至原肠期及孵出期的比例则与对照组无显著差异。胰酶质量浓度为2和1mg／mL时．融合卵发育到各阶段的比例均低于对照组，统计分析表明两者的差异是显著的（P〈0．05）。研究表明，溶液种类、胰酶浓度等对电融合卵的发育有明显的影响，但延缓受体卵的激活时间并不能提高电融合卵的出苗率。用郝氏液配制的2mg／mL的胰酶溶液处理受体卵，可获得较高比例的融合鱼。     

波吉卵囊藻对抗生素的敏感性及其无菌化培养

研究了青霉素、链霉素、庆大霉素和卡那霉素对波吉卵囊藻生长的影响,采用4种方法添加抗生素获得无菌藻株,比较波吉卵囊藻除菌前后的生长特点。结果表明:青霉素和链霉素浓度为1000IU/mL时,对波吉卵囊藻生长无明显影响;庆大霉素和卡那霉素浓度达500IU/mL时,对波吉卵囊藻生长无明显影响。微藻培养液中细菌对抗生素敏感性实验表明,除青霉素外,链霉素、庆大霉素和卡那霉素均对藻液中细菌有明显抑制作用。采用4种方法添加抗生素,均可获得无菌藻株。与未除菌时相比,无菌的波吉卵囊藻不易老化。     

苏氏圆腹胚胎发育的初步观察

观察两批苏氏圆腹受精卵的胚胎发育,描述并记录胚胎发育各期的形态特征及发育时间。结果表明:苏氏圆腹成熟卵呈圆球形,淡黄色,富有光泽,卵黄分布均匀,相对密度略大于水,卵径0.93～1.11 mm,平均0.98±0.19 mm。受精卵吸水后呈微黏性,属黏性卵,卵膜略膨胀,卵径0.99～1.15 mm,平均1.08±0.27 mm。苏氏圆腹胚胎发育属于盘状卵裂,与其他淡水硬骨鱼类基本相似。其发育过程可分为:受精卵、卵裂期、囊胚期、原肠期、神经胚期、器官分化期、出膜期和初孵仔鱼。在水温26±1℃的条件下,苏氏圆腹胚胎孵化历时24 h 45 min,总积温640.90 h.℃。初孵仔鱼体全长3.20～3.60 mm。     

栉江珧受精过程的细胞学研究

采用阴干、温差刺激诱导栉江珧亲贝排放和解剖亲贝获取精卵,授精后,通过连续取样直接镜检和卡诺液固定、Giemsa染色后观察,以研究栉江珧受精过程的细胞及染色体变化。结果表明,栉江珧成熟或接近成熟的卵,大多停留在第一次成熟分裂的前中期,接触海水后,进一步发育至中期。通常在26℃的海水中,授精后20min和35min,分别排出第一和第二极体。大约在授精后45min,雌雄原核融合,受精卵开始第一次卵裂。研究还表明,氨海水具有十分明显刺激卵内物质运动和促进卵细胞成熟的作用。     

Mg2+、Mn2+对波吉卵囊藻生长的影响

【目的】研究Mg²⁺、Mn²⁺对波吉卵囊藻生长和多糖代谢的影响。【方法】取蒸馏水培养数日后的波吉卵囊藻液接入以f/2培养液的配方为基础的新鲜培养液中,Mg²⁺浓度梯度分别设置为0、1、2、4、8 mg/L,Mn²⁺浓度梯度分别设置为0、0.005、0.050、0.500、5.000 mg/L,实验周期为10 d。【结果与结论】不同浓度Mg²⁺、Mn²⁺对波吉卵囊藻的生长有显著性影响,当Mg²⁺质量浓度为2 mg/L时,波吉卵囊藻的相对增长率、色素蛋白含量均出现最大值,显著高于Mg²⁺添加量为0、4和8 mg/L的组(P<0.05),当Mn²⁺质量浓度为0.005 mg/L时,波吉卵囊藻相对增长率、色素蛋白含量均出现最大值,显著高于Mn²⁺添加量0、0.050、0.500和5.000 mg/L组(P<0.05)。不同浓度Mg²⁺、Mn²⁺对波吉卵囊藻胞外多糖含量有显著影响(P<0.05),当Mg²⁺质量浓度>2 mg/L、Mn²⁺质量浓度>0.005 mg/L时,波吉卵囊藻生长均受到胁迫,其胞外多糖合成量明显增加,以保护藻体不受危害;当Mg²⁺质量浓度为8 mg/L、Mn²⁺质量浓度为5.000 mg/L时,波吉卵囊藻胞外多糖含量分别为对照组的2.43和3.36倍。     

双棘黄姑鱼血清性类固醇激素的周年变化

采用放射免疫法(RIA)测定双棘黄姑鱼(Nibea diacanthus)血清性类固醇激素的周年变化。结果表明:雌鱼血清雌二醇(Estadiol-17β)浓度一年出现两次峰值,均在成熟产卵前一个月,即4月[(338±59)pg/mL]和9月[(296±48)pg/mL];血清睾酮(T)浓度在卵黄发生期急剧上升,并在卵巢成熟期达到最高值,分别是5月(2.506±0.262 ng/mL)和10月份(2.268±0.243 ng/mL),分别在血清雌二醇浓度两次峰值出现1个月之后,因此,雌鱼的血清雌二醇浓度和血清睾酮浓度均与性腺发育相关。雄鱼血清睾酮浓度的两次峰值均出现在性腺已发育成熟的5月份(2.307±0.792 ng/mL)和10月份(2.286±0.607 ng/mL),与性腺发育有明显的相关性;而血清雌二醇浓度全年维持在一个相对稳定的水平,与雄性性腺发育相关性不明显。     

弓背青鳉的胚胎发育及自发荧光观察

【目的】研究弓背青鳉(Oryziascurvinotus)胚胎各阶段的发育特点,观察其自发荧光。【方法】使用正置荧光显微镜对弓背青鳉受精到孵化出膜过程进行连续观察。【结果】弓背青鳉胚胎发育历时10～12d,发育过程分为受精卵激活期、受精卵胚盘形成期、卵裂期、囊胚期、原肠胚期、神经胚期、器官形成期及孵化出膜期共8个时期;根据各时期胚盘形成、体节数量、器官发生、胚体大小和运动等特征的变化,进一步细分为35个阶段。其自发荧光主要开始出现于12肌节期,由虹彩细胞团内的物质在激发光下产生,主要分布于脑与脊索背面的皮肤表层。     

三角帆蚌肉酶解产物响应面优化制备及醒酒活性

【目的】研究三角帆蚌(Hyriopsis cumingii)肉酶解产物的解酒防醉作用。【方法】以乙醇脱氢酶(ADH)激活率为指标,在单因素试验基础上响应面优化三角帆蚌肉酶解工艺,制备三角帆蚌肉酶解产物(Enzymatic hydrolysate from soft tissue of Hyriopsis cumingii, EH),并测定EH对饮酒小鼠醉酒状态、血液中乙醇浓度和肝脏ADH活力的影响,以评价其醒酒活性。【结果】选用中性蛋白酶为实验用酶,三角帆蚌肉的最佳酶解条件为酶解时间2.0 h、酶解温度50.0℃、料液比1∶3、加酶量1 000 U/g,在此条件下ADH激活率为18.30%。小鼠摄入EH300 mg/kg时,饮酒小鼠醉酒只数少,酒后90 min血乙醇浓度已降至8.80 mmol/L,肝脏ADH活力上升至7.64 U/mg。【结论】三角帆蚌肉酶解产物能够显著激活肝脏ADH,降低血乙醇浓度,具有较好的醒酒功效和应用前景。     

ERR-dsRNA对罗氏沼虾卵巢中ERR及生殖相关基因表达的影响

【目的】研究雌激素相关受体(Estrogen-related receptor,ERR)在雌性罗氏沼虾生殖过程中的功能。【方法】通过注射双链RNA(Double-strandedRNA,dsRNA)干扰罗氏沼虾(Macrobrachium rosenbergii)ERR的表达,对干扰组和对照组的卵巢样本进行转录组测序,筛选与生殖相关的差异表达基因,并通过qPCR检测它们在卵巢发育过程中的表达。【结果】注射ERR-dsRNA后,卵巢中ERR mRNA和ERR蛋白的表达显著下降。ERR-dsRNA干扰组和对照组卵巢转录组测序获得的差异表达Unigene中,9条被富集到生殖GO条目,3条被富集到生殖过程GO条目,21条被富集到卵母细胞减数分裂Pathway条目。这些生殖相关的Unigene中差异倍数较大的有9条,经qPCR验证,5条在ERR干扰组和对照组间的表达差异有统计学意义。这5条Unigene中有3条在卵巢发育过程中的表达差异有统计学意义,其中2条(CL7112.Contig4_All和Unigene4600_All)与卵母细胞减数分裂有关,1条(CL4888.Contig2_All)与生殖相关激素的合成及释放有关。【结论】ERR可通过调控卵母细胞减数分裂和生殖相关激素的合成与释放调控罗氏沼虾卵巢发育。     

Hg-Cd复合胁迫对卵叶喜盐草光合色素和丙二醛含量及抗氧化酶活性的影响

通过胁迫试验研究Hg-Cd复合胁迫对卵叶喜盐草光合色素含量、丙二醛(MDA)含量及抗氧化酶活性的影响。结果表明,Hg-Cd复合毒性对卵叶喜盐草叶绿素a、叶绿素b和类胡萝含量显著影响(P0.05),当Hg、Cd质量浓度分别大于0.005和0.100 mg·L~(-1)时,叶绿素a、b比值,叶绿素a和类胡萝卜素含量下降,光合作用受抑制;Hg-Cd复合胁迫对卵叶喜盐草的MDA含量影响有统计学意义(P0.05),当Hg、Cd质量浓度分别为0.005和0.100 mg·L~(-1)时,MDA含量上升41.5%;Hg、Cd复合胁迫对卵叶喜盐草的超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性影响有统计学意义(P0.05),SOD活性随浓度梯度逐渐下降,在Hg、Cd质量浓度分别大于0.010和0.200 mg·L~(-1)时,SOD活性降低,POD和CAT活性随胁迫浓度梯度呈先增后减趋势,其中当Hg、Cd质量浓度分别为0.010和0.20 mg·L~(-1)时,而POD和CAT活性均上升为最大值,分别较对照组增加12.35%和22.75%,当Hg、Cd质量浓度分别大于0.015和0.300 mg·L~(-1)时,POD和CAT活性开始下降至最低。总的来说,Hg、Cd联合胁迫的浓度大于0.005和0.100 mg·L~(-1)时,卵叶喜盐草光合色素含量下降,MDA含量和抗氧化酶活性均相应发生变化。     

STUDY ON EMBRYONIC DEVELOPMENT AND EARLY GROWTH OF TRIPLOID AND GYNOGENETIC DIPLOID LEFI—EYED FLOUNDER, PARALICHTHYS OLIVACEUS （T. et S. ）

The early effects of chromosomal manipulation of eggs and sperm on the yields of trip-loid and gynogenefic diploid larvae of Paralichthys olivaceus were investigated. Triploidy was achieved by cold shocking fen.ilized eggs at 0 - 2℃ for 45 minutes duration 5 minutes after fen.ilization, and the in-duced triploidy rates were 31.2% - 50% and the relative hatching rates were 53.3% - 99%. Gynnge-aetic diploids were obtained when eggs were inseminated with irradiated sperm and cold shocked at 0 -2℃ for 45 minutes duration 5 minutes after fertihzation. The induced gynogenetic diploid rates and the relative hatching rates were 94 % - 96 % and 48.5 % - 68.5 % respectively. The embryonic development of the triploid experimental group and of the gynogenetic diploid experimental group was delayed at first compared with the control group. But from the gastrula stage, it was not delayed anymore. There were no significant differences in the growth of the triploid experimental group larvae and the control group larvae, and in the growth of the gynogenetic diploid experimental group larvae and the control group larvae ac-cording to Student‘s t-test (α = 0.05). The relationship between the early growth of the triploid experi-mental group larvae and that of gynogenetic diploid experimental group larvae was also studied.     

Morphological changes and allometric growth in hatcheryreared Chinese loach Paramisgurnus dabryanus(Dabry de Thiersant, 1872)

The length-weight relationship and allometric growth patterns of hatchery-reared Chinese loach, Paramisgurnus dabryanus(Dabry de Thiersant, 1872), were determined from hatching to 60 days after hatching. A best power model was estimated for the length-weight relationship during the early life stages. Positive allometric growth for the head segment, trunk length, tail length and eye diameter was also found in the early life stages, while body depth, tail depth, tail fi n length, pectoral fi n length and barbel length displayed a negative coeffi cient. During the subsequent early developmental stage, the growth coeffi cients showed a clear and common tendency towards isometry for all measured body ratios. The allometric growth changes in Chinese loach during the early stage are possibly the result of selective organogenesis directed towards survival priorities.     

BIOLOGICAL EXPERIMENT AND CLINICAL TESTS ON MAMMOTROPIC ACTION OF MUYINGLE -A NEW TYPE OF HEALTHFOOD PREPARED FROM MARINE ORGANISMS

Muyingle is a new type of health food prepared from marine organisme. The mammotropic action ofMuyingte was investigated by studying its effect on mammary glands and pituitary glands of lactating miceand the survival rate of suckling mice. The results showed that the mammotropic action of Muyingle wasvery effective.The survival rates of suckling mice were 92.90% for the treated group and 0 for the controlgroup (p<0.01). The weights of mammary gland were 163±51.1mg/10g (weight of mouse)for the treatedgroup and 98.5±18.4 mg/10g for the control group (p<0.01). Histological examinations suggestal thatmammary glands from the treated group were at the secreting stages, while those from the control groupwere at the resting stages. Clinical tests also demonstrated that Muyingle was highly effective in promotinglactation and improving the quality of the puerpera's milk. The efficiency was up to 86.7%.     

Morphological changes and allometric growth in hatchery-reared Chinese loach <Emphasis Type="Italic">Paramisgurnus dabryanus</Emphasis> (Dabry de Thiersant, 1872)

The length-weight relationship and allometric growth patterns of hatchery-reared Chinese loach, Paramisgurnus dabryanus (Dabry de Thiersant, 1872), were determined from hatching to 60 days after hatching. A best power model was estimated for the length-weight relationship during the early life stages. Positive allometric growth for the head segment, trunk length, tail length and eye diameter was also found in the early life stages, while body depth, tail depth, tail fin length, pectoral fin length and barbel length displayed a negative coefficient. During the subsequent early developmental stage, the growth coefficients showed a clear and common tendency towards isometry for all measured body ratios. The allometric growth changes in Chinese loach during the early stage are possibly the result of selective organogenesis directed towards survival priorities.     

饲料中添加L-肉碱对牙鲆幼鱼生长、生化组成及血液指标的影响

研究0、300、600、900、1 200、1 500 mg/kg 6个L-肉碱添加水平对牙鲆(Paralichthys olivaceus)幼鱼生长、生化组成和血液指标的影响。结果表明:添加量为1 200 mg/kg的处理组牙鲆幼鱼的增重率和特定生长率显著提高,饲料系数显著降低(P<0.05);随着饲料中L-肉碱含量的增加,肝指数呈现下降趋势,且在添加量为900~1 500 mg/kg时,各组较对照组差异显著(P<0.05);饲料中添加L-肉碱显著降低各实验组脏体比(P<0.05),而对各处理组间存活率和肥满度均无影响(P<0.05);肌肉和肝脏中的脂肪含量与L-肉碱添加量呈负相关,当添加量为1 200~1 500 mg/kg时,较对照组差异显著(P<0.05);肌肉中的蛋白含量与L-肉碱添加量呈正相关,但各组差异不显著(P<0.05);饲料中添加L-肉碱降低血清中的谷草转氨酶、胆固醇、甘油三酯和尿素氮含量,且添加量1 200 mg/kg组各指标含量均显著低于对照组(P<0.05);各处理组的血糖含量较对照组均有所增加,其中1 200 mg/kg组血糖含量较对照组提高41.6%(P<0.05);血清中总蛋白和低密度脂蛋白均随着L-肉碱添加量的增加而呈下降趋势,高密度脂蛋白却得以提升,但差异均不显著(P<0.05)。分析认为,在本实验条件下,饲料中添加L-肉碱可促进牙鲆幼鱼生长,且降脂效果显著。建议L-肉碱最适添加量为1 200 mg/kg。     

水产饲料中三聚氰胺检测方法的优化

通过对高效液相色谱法(HPLC)测定水产饲料中三聚氰胺含量的方法进行优化,得到最佳色谱条件和前处理方法,建立了水产饲料中三聚氰胺的检测方法。结果表明:以体积比8∶92的乙腈、庚烷磺酸钠离子对试剂为流动相,柱温40℃,流速1.2 mL/min,以波长236 nm检测;以提取液为18 mL50%(φ)乙腈水+1 mL乙酸锌+1 mL1mol/L盐酸,淋洗液3 mL水和3 mL甲醇,洗脱体积6 mL作样品前处理。三聚氰胺加标回收率为85%～100%,相对标准偏差(RSD)小于5%,线性范围为0.2 mg/L～5.0mg/L,相关系数0.999 8,方法检出限为0.2 mg/kg。     

马氏珠母贝育珠细胞小片和珠核处理技术研究Ⅰ:不同处理液在海南陵水育珠的效果

研究8组不同小片和核处理液在海南陵水海区的育珠效果,优化、筛选适宜该海区育珠的小片处理液及与之匹配的珠核处理液。结果显示:1)处理组手术贝休养期的吐核率与死亡率均显著低于对照组,表明处理液可促进伤口愈合并抗感染;2)处理组育珠8个月后的珠层厚度平均可达0.5 mm以上,与对照组0.3 mm的珠层厚度差异极显著(P<0.01),表明处理液具有明显促进珍珠质分泌的作用;3)优化筛选出核处理液C3,其增强核亲和力的效果明显优于C1(P<0.05);4)W处理组育珠贝死亡率平均小于22%,明显低于对照组(36.7%)和Y处理组(31.45%)(P<0.01),表明W处理液效果优于Y处理液;5)在8个处理组中,筛选出W0、W2、Y3三个匹配组,其育珠效果明显优于对照组(P<0.01)。引入综合评价指数即优良珍珠数占育珠贝总数的比例,以量化育珠效果便于比较     

Ontogenetic changes in digestive enzyme activities and the amino acid profile of starry flounder Platichthys stellatus

Ontogenetic changes in digestive enzyme activities and the amino acid(AA) profile of starry flounder,Platichthys stellatus,were investigated and limiting amino acids were estimated compared with the essential AA profile between larvae and live food to clarify starry flounder larval nutritional requirements.Larvae were collected at the egg stage and 0,2,4,7,12,17,24 days after hatching(DAH)for analysis.Larvae grew from 1.91 mm at hatching to 12.13 mm at 24 DAH.Trypsin and chymotrypsin activities changed slightly by 4 DAH and then increased significantly 4 DAH.Pepsin activity increased sharply beginning 17 DAH.Lipase activity increased significantly 4 DAH and increased progressively with larval growth.Amylase activity was also detected in newly hatched larvae and increased 7 DAH followed by a gradual decrease.High free amino acid(FAA) content was detected in starry flounder eggs(110.72mg/g dry weight).Total FAA content dropped to 43.29 mg/g in 4-DAH larvae and then decreased gradually to 13.74 mg/g in 24-DAH larvae.Most FAAs(except lysine and methionine) decreased 50%in 4-DAH larvae compared with those in eggs and then decreased to the lowest values in 24-DAH larvae.Changes in the protein amino acid(PAA) profile were much milder than those observed for FAAs.Most PAAs increased gradually during larval development,except lysine and phenylalanine.The percentages of free threonine,valine,isoleucine,and leucine decreased until the end of the trial,whereas the protein forms of these four AAs followed the opposite trend.A comparison of the essential AA composition of live food(rotifers,Artemia nauplii,and Artemia metanauplii) and larvae suggested that methionine was potentially the first limiting AA.These results may help develop starry flounder larviculture methods by solving the AA imbalance in live food.Moreover,the increased digestive enzyme activities indicate the possibility of introducing artificial compound feed.     

波纹龙虾胚胎的离体培养及发育观察

对波纹龙虾不同发育时期的胚胎进行离体培养和显微观察。结果表明,初始培育的离体胚胎愈接近孵化期,其孵化率愈高(最高达80.0%);波纹龙虾胚胎发育经历受精卵、卵裂期、囊胚期、原肠期、中眼色素形成期、复眼色素形成期、心跳期、破膜前期和出膜期;胚胎在发育过程中颜色变化:橙红→深橙红→砖红→灰白→接近透明。     

食物类型和浓度对双齿许水蚤摄食的影响

在不同饵料条件下,对双齿许水蚤(Schmackeria dubin)的清滤率(mL.只-1.h-1)、滤食率(细胞.只-1.h-1)和摄食率(mg.只-1.h-1)进行了测定。结果表明:以湛江等鞭金藻(Isochrysis zhanjiangensis)为饵料,其浓度为15×104～120×104 mL-1时,双齿许水蚤的摄食率达到较大;分别饲以同生物量的牟氏角毛藻(Chaetoceros müelleri)、亚心形扁藻(Platymonas subcordiformis)、湛江等鞭金藻、小球藻(Chlorella sp.)时,饵料为角毛藻和扁藻时双齿许水蚤的摄食率达到较大;饲以同生物量的牟氏角毛藻、亚心形扁藻、湛江等鞭金藻、小球藻、颤藻(Oscillatoria sp.)、小环藻(Cyclotella sp.)六种藻的混合饵料时,双齿许水蚤对湛江等鞭金藻的摄食率和选食率均为最大,分别为0.0458 mg.只-1.h-1和17.1%。可见,饵料浓度、饵料种类、混合饵料对双齿许水蚤的清滤率和摄食率均有显著影响(P<0.05)。