17α-甲基睾酮对赤点石斑鱼性逆转的影响

采用腹部埋植方式,用17α-甲基睾酮(17-αMT)以10mg.kg-1剂量处理2龄雌性赤点石斑鱼(每4周埋植1次,共埋植2次),检查埋植前后性腺结构、血清中性类固醇激素水平以及脑和性腺芳香化酶活性的变化。结果显示:一次埋植17-αMT即可诱导赤点石斑鱼发生不同程度的性逆转,性腺成熟系数(GSI)明显下降,性腺中卵细胞退化,精原细胞增殖,出现大量精母细胞和精子细胞,性逆转雄鱼精巢的组织结构与正常雄鱼精巢相似,部分鱼转变为功能性雄鱼,经轻微的腹部挤压可排精,精子活力与正常雄鱼相似。赤点石斑鱼埋植17-αMT后前脑、中脑的芳香化酶活性明显提高而后脑的芳香化酶活性降低,但性腺芳香化酶活性无显著变化。埋植17α-MT后第4周血清中11-酮基睾酮(11-ketotestosterone,11-KT)浓度显著升高,而睾酮(testosterone,T)和雌二醇(estradiol-17,βE2)浓度变化不显著。这些结果表明,脑部芳香化酶活性的显著升高与性逆转密切相关,17-αMT主要通过提高血清中11-KT水平诱导赤点石斑鱼发生性逆转。     

4种硝基呋喃类代谢物在青石斑鱼肌肉中的富集与消除规律

实验通过连续7 d投喂各含有100 mg·kg~(-1)呋喃西林、呋喃唑酮、呋喃它酮和呋喃妥因的饲料,研究4种硝基呋喃药物在青石斑鱼(Epinephelus awoara)肌肉中的消除规律。采用超高效液相色谱串联质谱(UPLCMS/MS)检测青石斑鱼肌肉中的氨基脲(SEM)、3-氨基-2-恶唑基酮(AOZ)、5-甲基吗啉-3-氨基-2-唑烷基酮(AMOZ)和1-氨基-2-内酰脲(AHD)的质量分数,并用内标法定量。结果表明,SEM、AOZ、AMOZ和AHD在石斑鱼背肌的峰浓度最大值依次为1 104.10μg·kg~(-1)、781.09μg·kg~(-1)、679.75μg·kg~(-1)和437.18μg·kg~(-1)。4种硝基呋喃代谢物在石斑鱼肌肉组织中的消除规律大致相似。SEM、AHD、AOZ和AMOZ半衰期时间的长短顺序为23.90 d、23.10 d、22.35 d和21.00 d。SEM在肌肉中消除最慢。消除到第163天时,SEM、AOZ、AMOZ和AHD在肌肉中的质量分数仍然高于液质检出限(0.5μg·kg~(-1))。建议硝基呋喃药物应禁止用于石斑鱼疾病的预防或治疗中。     

饲料维生素E水平对云纹石斑鱼幼鱼生长、营养及免疫指标的影响

以初始体重(15.58±0.22)g的云纹石斑鱼(Epinehelus moara)为研究对象,设置5种不同维生素E水平(11.09 mg·kg-1、47.52 mg·kg-1、91.38 mg·kg-1、134.57 mg·kg-1、178.92 mg·kg-1)的等氮等能饲料,对云纹石斑鱼进行56 d的饲育实验,探讨饲料维生素E水平对该鱼生长、营养性能及血清免疫指标的影响。结果表明:饲料中添加维生素E对云纹石斑鱼幼鱼的增重率、特定生长率和饲料效率的作用明显,饲料维生素E在149.59 mg·kg-1左右时云纹石斑鱼的增重率较高;云纹石斑鱼组织中维生素E积累量与饲料中维生素E添加水平呈正比,但饲料维生素E水平超过91.38 mg·kg-1时鱼体组织中维生素E积累量不随饲料维生素E水平添加而显著升高,饲料维生素E水平在139.68 mg·kg-1时云纹石斑鱼具有较高的肝脏维生素E积累量;饲料维生素E水平对云纹石斑鱼血清中免疫球蛋白M和溶菌酶活性的影响显著。综合增重率、肝脏维生素E积累量和免疫指标,云纹石斑鱼饲料维生素E适宜添加水平为139.68~149.59 mg·kg-1。     

药浴暴露下阿维菌素在异育银鲫体内蓄积和消除规律

采用UPLC-MS/MS法研究了2μg·L-1三次连续水体药浴和6μg·L-1一次性水体药浴条件下阿维菌素在水体中消除、在异育银鲫(Carassius auratus gibelio)体内蓄积和消除变化规律。结果显示,两种药浴暴露方式下阿维菌素在水体中消除呈一级指数衰退消除,消除半衰期(t1/2)均为63 h,240 h时浓度下降到0.5μg·L-1以下。阿维菌素在异育银鲫血浆和肌肉中的含量均呈先升高后下降的趋势,血浆中药物浓度远高于肌肉中的含量。2μg·L-1连续三次药浴组和6μg·L-1一次药浴组血药峰浓度(Cmax)分别为34.97、66.62μg·L-1,其曲线下面积(AUC0-t)分别为9 871.2μg·L-1·h和18 119.6μg·L-1·h;两组药浴肌肉中达峰浓度分别4.42μg·kg-1和15.80μg·kg-1,其AUC0-t分别为641.9μg·kg-1·h和4 271.0μg·kg-1·h。与2μg·L-1连续三次药浴组相比,6μg·L-1一次药浴组阿维菌素在血浆和肌肉中的蓄积作用更加显著。以10μg·kg-1作为阿维菌素在异育银鲫肌肉中最大残留限量,选择95%的置信区间计算异育银鲫肌肉组织中休药期,本研究中2μg·L-1连续三次药浴组肌肉的休药期为295.4 h,6μg·L-1一次药浴组肌肉的休药期为454.5 h。     

循环水养殖赤点石斑鱼幼鱼放养密度研究

将体质量(10.82±0.17)g的赤点石斑鱼(Epinephelus akaara)幼鱼饲养在室内循环水、直径40cm×水深50cm池中,每个池中10(G_(10))、20(G_(20))和40(G_(40))尾,投喂常规饲料,每周测4次水质。8周的养殖结果表明,养殖时间和放养密度均影响赤点石斑鱼的生长、存活和水质。随着养殖时间的加长,成活率逐步降低,但整体保持在40.00%~90.00%之间,G_(10)组的成活率显著高于G_(40)组(P0.05)。试验结束时,赤点石斑鱼的增重率(WGR)变化在(47.97±1.98)~(68.02±2.34)%之间,特定生长率(SGR)变化在(0.70±0.33)~(0.93±0.42)%/d之间,两指标不同密度组差异显著(P0.05),而各组鱼的肝体比(HIS)、内脏比(VSI)和肥满度(CF)差异不显著(P0.05)。     

室外池塘自然养殖条件下呋喃西林代谢物在中华绒螯蟹体内残留和消除规律

采用高效液相色谱串联质谱(HPLC-ESI/MS/MS)法分析呋喃西林主要代谢物氨基脲(Semicarbazide,SEM)在中华绒螯蟹(Eriocheir sinensis,以下简称河蟹)体内的残留及消除规律。设2个试验组和1个对照组,2个试验组以20 mg·L-1和80 mg·L-1的呋喃西林溶液浸泡河蟹(扣蟹)60 min后,转移至饲养池塘中,并在给药后1 h、2 h、4 h、8 h、12 h、24 h、2 d、4 d、8 d、12 d、16 d、20 d、30 d、40 d、50 d、60 d、80 d、100 d、120 d、150 d进行各组织取样分析。结果显示:在停药2 h时,2个浓度试验组河蟹的肌肉、肝胰腺和鳃中SEM均达到高峰,其中,20 mg·L-1试验组测定值分别为(152±21.7)、(234.0±12.0)、(3 160±169)μg·kg-1,80 mg·L-1试验组测定值分别为(327±31.2)、(372±27.2)、(4 623±247)μg·kg-1。停药2 h后,2个浓度试验组河蟹各组织中SEM均呈现不同的消除速率,其中20 mg·L-1试验组的消除速率分别为0.315μg·(kg·h)~(-1)(肌肉)、0.487μg·(kg·h)~(-1)(肝胰腺)和4.39μg·(kg·h)~(-1)(鳃);80 mg·L-1试验组的消除速率分别为0.454μg·(kg·h)~(-1)(肌肉)、0.516μg·(kg·h)~(-1)(肝胰腺)和6.43μg·(kg·h)~(-1)(鳃)。停药960 h后,2个浓度试验组各组织中SEM残留均低于判定限(1.0μg·kg-1)。     

地衣芽孢杆菌对尖吻鲈血液生理生化指标的影响

以初始体重为(17.47±0.19)g的尖吻鲈(Lates calarifer)为试验对象,在基础饲料中分别添加0(对照)、1.0、2.0、3.0、4.0、5.0和6.0 g·kg-1饲料的地衣芽孢杆菌(Bacillus licheniformis)粉状制剂,研究地衣芽孢杆菌对尖吻鲈的生理生化指标的影响.摄食不同地衣芽孢杆菌含量的尖吻鲈血液中红细胞和白细胞数量、血红蛋白和无机盐离子等没有显著性差异(P>0.05),但显著降低了尖吻鲈的血糖和尿素氮水平(P<0.05),血糖在2.0 g·kg-1饲料组达到最小值,尿素氮在1.0 g·kg-1饲料组达到最小值.血清总蛋白均高于对照组,并在4.0 g·kg-1饲料组显著升高(P<0.05);胆固醇除了3.0 g·kg-1饲料组显著低于对照组外,其他各试验组均与对照组差异不显著.地衣芽孢杆菌也降低了血清谷丙转氨酶及乳酸脱氢酶的活性,谷丙转氨酶在1.0、4.0、5.0和6.0 g·kg-1饲料组显著低于对照组(P<0.05),而乳酸脱氢酶只在1.0 g·kg-1饲料组显著低于对照组(P<0.05).     

不同免疫增强剂对仿刺参肠道消化酶活性及组织结构的影响

在15±0.5℃条件下,向初始体质量为(21.00±2.00)g的仿刺参(Apostichopus japonicas Selenka)体内注射400μL(质量浓度为1.2 mg·mL-1)几种免疫增强剂(黄芪多糖、枸杞多糖、黄柏提取物、苦参提取物、党参提取物、菊粉、天蚕素、神曲提取物、山楂提取物),第1、3、5d时测定仿刺参肠道消化酶活性,第5d时制作前肠组织切片,研究不同免疫增强剂对仿刺参肠道消化酶活性及组织结构的影响。结果表明,在注射免疫增强剂后第3d,菊粉组仿刺参肠道蛋白酶活性最高,达15.16μg·g-1·min-1,与对照组7.07μg·g-1·min-1差异显著(P<0.05),而黄柏提取物抑制了蛋白酶活性;神曲组淀粉酶活性达72.57 U·dl-1,与对照组(53.2 U·dl-1)差异显著(P<0.05);菊粉和山楂提取物对仿刺参肠道纤维素酶活性影响显著(P<0.05),其他处理组与对照比较无明显变化(P>0.05);黄芪多糖组褐藻酸酶活性最高,达2.06μg·g-1·min-1,其次是菊粉组。在养殖试验期间淀粉酶比活力变化趋势较稳定,纤维素酶次之,褐藻酸酶和蛋白酶变化幅度较大。各种免疫增强剂对仿刺参肠道组织结构的影响不同,黄柏、苦参提取物损伤了组织结构,而黄芪多糖、菊粉等促进了细胞分泌。     

杂交种“金虎石斑鱼”与棕点石斑鱼卵巢发育及性类固醇激素水平比较

为了探究杂交种“金虎石斑鱼”(Epinephelus fuscoguttatus ♀×Epinephelus tukula ♂)和母本棕点石斑鱼(Epinephelus fuscoguttatus)的卵巢发育和血清中性类固醇激素水平的变化,对10、18、24、36月龄的金虎石斑鱼和棕点石斑鱼进行了卵巢组织切片,并采用酶联免疫吸附法测定其血清中的雌二醇(E2)、11-酮基睾酮(11-KT)、睾酮(T)、孕酮(P)含量。结果显示,杂交种金虎石斑鱼卵巢正常发育,没有出现卵母细胞萎缩、退化的现象,且36月龄时金虎石斑鱼卵巢发育至Ⅲ期早期,卵母细胞进入大生长期体积明显增大,核仁多位于核膜外侧。棕点石斑鱼卵巢发育较快,36月龄时卵巢已发育至Ⅲ期末期,卵母细胞与核体积进一步增大。卵膜外又形成了一层滤泡膜,形成双层滤泡膜结构。金虎石斑鱼与棕点石斑鱼血清中E2、11-KT、P的水平变化趋势相似,呈现先升后降的趋势,且在24月龄时达到最大,但棕点石斑鱼高于金虎石斑鱼。金虎石斑鱼血清中T的水平与其他性类固醇激素变化一致:在24月龄时达到最大,在36月龄时显著下降,而棕点石斑鱼血清中T在36月龄时依然维持和2...     

胸腺肽对AA肉鸡血液常规指标的影响

目的研究胸腺肽对AA肉鸡血液常规指标的影响。方法将125只14日龄健康AA肉鸡随机分为五个组,每组25只,其中4个试验组(a、b、c、d组)分别按50μg/kg、150μg/kg、300μg/kg、500μg/kg剂量,肌肉注射胸腺肽,阴性对照组(e组)同样的方式和剂量注射生理盐水,1次/d,连续注射5 d。分别于用药前(14日龄)、用药5 d(19日龄)及停药5 d(24日龄)对各组肉鸡进行血液常规指标的测定。结果连续5 d肌肉注射,各实验组的肉鸡淋巴细胞的百分比显著升高,中性粒细胞百分比显著降低;d组(500μg/kg)肉鸡的白细胞总数、嗜碱性粒细胞百分比较对照组显著增高,淋巴细胞百分比差异不显著。停药后5 d,各组肉鸡的血液生理指标除红细胞数量减少外,其他均趋于用药前水平。结论注射500μg/kg以下各剂量时,胸腺肽可能增强AA肉鸡的免疫力,对其他各项血常规指标没有明显的不良影响,可在临床上推广使用。     

甲壳动物CHH家族神经激素结构和功能研究进展

甲壳动物主要利用温度、光照周期等外界因子调节其生理状态 ,使它们的生殖活动处于最适条件下 ,来自外界因子的这些信息作用于甲壳动物的中枢神经系统 ,后者将其传递到神经内分泌系统和内分泌系统 ,神经内分泌系统和内分泌系统能够分泌一些促进因子和抑制因子以实施对性腺活动的调控。由于甲壳动物成体的生殖和蜕皮常常交替出现 ,因此 ,神经内分泌系统和内分泌系统的精确调控非常重要。甲壳动物高血糖激素 (ＣＨＨ)家族神经激素是甲壳动物特有的多肽激素 ,它们主要由眼柄的Ｘ -器官窦腺复合体合成 ,它们包括 :甲壳动物高血糖激素 (ＣＨＨ)、…     

Enhanced survival in striped bass fingerlings after maternal triiodothyronine treatment

Elevation of the triiodothyronine (T₃) content of striped bass (Morone saxatilis) eggs by maternal T₃ injection confirms the uptake of T₃ by oocytes. The resulting offspring were influenced favorably by the T₃, as seen in quantitative indices of development. As reported previously, larvae from T₃-supplemented eggs raised under laboratory conditions exhibited increased body area, length, dry weight, and rates of swimbladder inflation and survival, compared to controls. Also, the T₃ content of unfertilized oocytes correlated positively and highly significantly with survival to two weeks of age within individual cohorts (Brownet al., 1988). In the present study, the survival of experimental and control striped bass was monitored through the fingerling stage, under hatchery production conditions. The rate of recovery of maternally T₃-treated cohorts from pond-culture was approximately fourfold that of controls. The striking effects of T₃ enrichment of eggs on offspring indicate the potential contribution of maternal hormones in striped bass development, and suggest possible applications in aquaculture.     

Effects of 11-ketotestosterone and gonadotropin-releasing hormone on follicle-stimulating hormone and luteinizing hormone gene expression in castrated and sham-operated male red seabream <Emphasis Type="Italic">Pagrus major</Emphasis>

ABSTRACT:   In order to clarify the roles of androgen and gonadotropin-releasing hormone (GnRH) on gonadotropin (GTH; luteinizing hormone [LH] and follicle stimulating hormone [FSH]) synthesis, effects of castration and implantation of GnRH analog (GnRHa) or 11-ketotestosterone (11-KT) on expression of GTH subunit, α-glycoprotein subunit (αGSU), FSHβ, and LHβ genes, during the early spermatogenic stage in male red seabream Pagrus major were examined. Male red seabream underwent castration or sham-operation and were subsequently implanted with cholesterol pellets containing GnRHa, silicone capsules filled with 11-KT, or blank capsules (control). FSHβ mRNA levels increased due to castration, and it was reversed by treatment with 11-KT. 11-ketotestosterone treatment also decreased FSHβ mRNA levels in sham-operated fish. These results suggest that 11-KT acts on the pituitary to suppress FSH synthesis in male red seabream. On the other hand, neither castration nor replacement of 11-KT in castrated fish had effects on LHβ mRNA levels, whereas 11-KT treatment had slightly but significantly decreased LHβ mRNA in sham-operated fish. αGSU mRNA levels were not changed by castration or 11-KT treatment in both sham-operated and castrated fish. Meanwhile, treatment with GnRHa significantly decreased FSHβ mRNA levels in sham-operated fish, but not in castrated fish. This suggests that GnRHa may down-regulate expression of FSHβ mRNA through the production of 11-KT in testis. LHβ and αGSU mRNA levels in sham-operated fish, but not in castrated fish, were significantly elevated by treatment with GnRHa.     

Interaction of orexin/hypocretin-like immunoreactive neurons with melanin-concentrating hormone and α-melanocyte-stimulating hormone neurons in brain of a pleuronectiform fish, barfin flounder

Immunohistochemical localization of orexin/hypocretin in the brain of a pleuronectiform fish, the barfin flounder Verasper moseri was examined as the first step in unraveling the possible function of the hormone in the brain. Orexin-A-like immunoreactive (ir) cell bodies were found to be located in the nucleus posterioris periventricularis (NPPv) of the hypothalamus, and orexin-A-like-ir fibers were detected not only in the hypothalamus but also extensively throughout the brain. The orexin-A-like-ir cell bodies did not project their fibers to the pituitary gland. Since melaninconcentrating hormone (MCH) and α-melanocyte-stimulating hormone (α-MSH) are suggested to regulate food intake in addition to orexin/hypocretin in the teleost fish, it was examined whether neural connections exist between orexin neurons and the MCH and α-MSH neurons in the barfin flounder brain by using double-staining immunohistochemistry. Some orexin-A-like-ir fibers were in close contact with the MCH-ir and α-MSH-ir cell bodies in the hypothalamus. Moreover, a few MCH-ir and α-MSH-ir fibers were in close contact with the orexin-A-like-ir cell bodies in the hypothalamus. These results suggest that reciprocal connections exist between the orexin and MCH neurons and between the orexin and α-MSH neurons in the brain of the barfin flounder.     

鱼类促性腺激素抑制激素及其受体的研究进展

促性腺激素抑制激素是2000年由日本学者首次从鹌鹑脑中分离出的一种新型下丘脑神经肽,通过其受体介导参与机体的生长、生殖以及摄食等生理过程。迄今,只在金鱼、斑马鱼、星点东方鲀、罗非鱼以及斜带石斑鱼等几种鱼中鉴定出促性腺激素抑制激素。目前,鱼类促性腺激素抑制激素的生理学功能研究相对较少,且存在争议。鱼类促性腺激素抑制激素及其受体的表达调控以及其他生理学功能仍有待进一步研究。本研究简要总结鱼类促性腺激素抑制激素及其受体的研究进展,并对促性腺激素抑制激素的生理学功能进行概括讨论,旨在加深对鱼类促性腺激素抑制激素的认识和了解,为进一步研究做铺垫。     

三苯基锡对褐菖鮋卵巢发育的影响

尽管三苯基锡(triphenyltin,TPT)与三丁基锡(tributyltin,TBT)在海洋环境中经常同时存在,但相对于TBT,对TPT的毒性研究还较少。本实验以海洋经济鱼类褐菖鲉(Sebastiscus marmoratus)为对象,研究环境水平TPT对卵巢发育的影响。结果发现,TPT抑制了卵巢发育,表现为性腺指数的降低。TPT还诱导卵巢总脂质水平的增加,而卵巢脂质蓄积可能影响卵巢的正常生殖功能。另外,TPT暴露后,卵巢游离睾酮水平增加,卵巢游离17β-雌二醇水平降低;同时,血清三碘甲腺原氨酸水平降低。因此,TPT对性激素和甲状腺激素水平的影响可能是其抑制卵巢发育的重要机制。实验结果为全面了解TPT产生鱼类生殖毒性效应、途径和机制提供了新的重要的资料。     

黑鲷生长激素及其受体的季节变化

采用黑鲷生长激素(seabream growth hormone,brGH)放射免疫测定法(brGHRIA)及黑鲷生长激素放射受体测定法(brGHRRA)，研究了不同季节(3月、6月、9月及12月)黑鲷血清GH水平及肝脏和性腺中生长激素 受体(growth hormone receptor,GHR)水平的变化，同时测定了黑鲷白肌及肝脏RNA-DNA比率及黑鲷肥满度 (condition factor,K)和性腺指数(gonadosomatic     

Changes in the immunostaining intensities of follicle-stimulating hormone and luteinizing hormone during ovarian maturation in the female Japanese flounder

The role of gonadotropin (GTH) in the reproduction of the Japanese flounder, Paralichthys olivaceus, was studied by assessing the changes in the apparent activity of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in the pituitary gland during gonadal maturation by immunohistochemical analyses. Corresponding changes in plasma levels of testosterone (T), estradiol-17β (E₂), and 17α,20β-dihydroxy-4–pregnen-3-one (DHP) were also studied. Reared fish at the early spawning to termination stages were sampled from May to August and wild fish at the previtellogenic to termination stages were caught at 3- to 4-week intervals between April and September offshore from the northern mainland of Japan by gill nets. The gonadosomatic index of the reared fish decreased from the early spawning stage to the termination stage, while that of the wild fish increased significantly from the previtellogenic stage to the early spawning stage and decreased thereafter. In the reared fish, the immunostaining intensities of FSH and LH were high during the spawning period, accompanied by high plasma levels of T, E₂, and DHP. In the wild fish, the immunostaining intensities of FSH and LH were low during the previtellogenic stage but increased during the maturing and spawning stages. These results indicate that both FSH and LH are likely associated with oocyte maturation in the Japanese flounder.     

半滑舌鳎生长激素及其受体基因的原核表达

根据已克隆的半滑舌鳎(Cynoglossus semilaevis)生长激素(GH)基因和生长激素受体Ⅰ(GHR-Ⅰ)基因的cDNA序列信息设计引物,用于扩增半滑舌鳎GH、GHR-Ⅰ基因序列。随后,分别利用表达载体pET-32a和pGEX-6P-1成功构建了包含半滑舌鳎GH、GHR-Ⅰ基因ORF全序列及不同ORF片段的6个重组质粒,并将获得的重组质粒在表达宿主菌大肠杆菌(Escherichia coli)BL21(DE3)中,用IPTG诱导表达。SDS-PAGE电泳结果显示:重组GH的pET-32a-CSGH质粒在40 kD处有特异性的蛋白条带,重组GHR-Ⅰ胞内区的pGEX-6P-1-CSGHR1-2质粒在70 kD处发现诱导的蛋白条带,其表达量分别占细胞蛋白总量的37.6%和20.2%;而构建的pGEX-6P-1-CSGH重组质粒和含有GHR-Ⅰ基因跨膜区的重组质粒(pET-32a-CSGHR1-1、pET-32a-CSGHR1-2和pGEX-6P-1-CSGHR1-1)均未获得表达产物。由此可见,半滑舌鳎GHR-Ⅰ基因的跨膜区可能影响其在大肠杆菌中的表达。研究结果为实现半滑舌鳎GH、GHR-Ⅰ制剂的开发及应用于养殖生产实践奠定基础。