虹鳟性腺分化的研究

对新安江人工养殖虹鳟性腺的自然分化进行了系统组织学研究。刚孵化出膜仔鱼的生殖嵴巳形成,原始生殖细胞成单行排列于生殖嵴上,并正在进行有丝分裂。水温10—11℃时,孵出后6—8周仔鱼的性腺出现解剖学分化。紧接着卵巢中的卵原细胞开始向初级卵母细胞过渡。精巢中的精原细胞向初级精母细胞过渡的时间比卵巢约晚2—3周。性腺的细胞学分化时间,雌雄鱼表现不一致。     

外源性类固醇对虹鳟稚鱼性腺发育和性别分化影响的组织学观察

本文报导了应用外源性类固醇激素:甲基睾酮和雌酮投喂虹鳟稚鱼对其性腺发育和性别分化影响的光显观察结果。利用激素诱导性反转,培育全雌虹鳟种群的研究,对虹鳟鱼养殖生产有一定实践意义。     

不同倍性长牡蛎闭壳肌和精子的组织学和超微结构观察

以二倍体、三倍体、四倍体长牡蛎(Crassostrea gigas)闭壳肌和精子为研究对象, 采用石蜡组织切片、透射电镜和扫描电镜方法, 比较了 3 种倍性长牡蛎闭壳肌和精子的组织学特性及超微结构的差异。研究结果显示, 二、三、 四倍体闭壳肌组织的肌纤维都包含粗肌丝、细肌丝、肌膜、肌束膜、肌内膜、线粒体、囊泡、小管、细胞核、肌浆网等结构, 但三者的形态及超微结构不同, 其中三倍体闭壳肌线粒体直径、肌纤维直径、粗肌丝密度显著大于二倍体和四倍体, 肌纤维密度显著小于二倍体和四倍体。不同倍性精子的扫描电镜和透射电镜结果显示, 二、三、四倍体长牡蛎精子在组成结构上相似, 分别由头部、中段和尾部构成, 但不同倍性精子中各结构大小和线粒体数目不同。在长牡蛎精子头部长度、宽度, 顶体高度、宽度, 线粒体长度的测量分析中, 四倍体精子的头部、顶体以及线粒体显著大于二、三倍体。三倍体精子的头部和顶体显著大于二倍体, 但三倍体精子的线粒体大小与二倍体相比无显著差异。此外, 二、三倍体精子均含 4 个线粒体, 但在本研究中发现了二倍体精子含有 5 个(占比 1%)线粒体的特例。四倍体长牡蛎精子的线粒体数目不同, 含有 4 个(占比 46%)、5 个(占比 52%)、6 个(占比 2%)这 3 种情况。 该研究阐明了不同倍性长牡蛎闭壳肌及精子的组织学特性和超微结构, 为深入研究长牡蛎肌纤维生物学特性和精子繁育生物学提供了基础资料。     

水温对虹鳟四种血清酶活性影响的比较研究

使用DS-2000全自动化分析仪,测定水温15℃、16．5℃、18．5℃和22℃时虹鳟血清酶活性。结果表明：谷氨酸脱氢酶（GLDH）活性在水温15℃～16．5℃小比率降低,16℃～18．5℃大比率升高,18．5℃～22℃较大比率降低;a-羟丁酸脱氨酶（a-HBDH）活性在水温15℃～16．5℃小比率降低,16,5℃～18．5℃较大比率增加,18．5℃～22℃较大比率降低;脂肪酶（LPS）活性在水温15℃～16．5℃小比率升高,16．5℃～18．5℃小比率降低,18．5℃～22℃小比率升高;淀粉酶（AIMS）活性在水温15℃～16．5℃较小比率降低,16．5℃～22℃小比率升高。本文为揭示虹鳟热致死及耐热机理提供有关数据,为冷水性鱼类虹鳟温水驯化养殖提供基础技术资料。     

虹鳟源嗜冷黄杆菌的分离鉴定及致病性研究

为确定患病虹鳟的病原,本实验从患病鱼溃烂肌肉中分离到2株细菌,分别命名为CH06和CH07,经回归感染证实分离菌株为导致此次虹鳟患病的病原菌,并进一步对其形态特征、理化特性、分子特征、血清型及耐药性进行分析。结果显示,CH06和CH07株在TYES琼脂平板上呈煎蛋状外观,产黄色素,氧化酶和过氧化氢酶呈阳性,能水解明胶和酪蛋白,不能水解淀粉,不能利用果糖、半乳糖和七叶苷等。16S rRNA比对结果显示,CH06和CH07株与嗜冷黄杆菌模式株NBRC 15942的同源性分别为99.35%和99.42%。综合菌株理化和分子特性确定CH06和CH07株为嗜冷黄杆菌。利用多重PCR方法鉴定CH06和CH07株的血清型均为1型(Fd型);多位点序列分型(MLST)分析表明,CH06和CH07株的基因型分别为ST-12和ST-78型,且均属于CC-ST10克隆型。人工感染结果显示,CH06和CH07株对虹鳟幼鱼具有较高致病性,其半致死浓度(LD₅₀)分别为7.1×10⁵和1.1×10⁵ CFU/mL,攻毒剂量与临床病症出现时间呈反比,从人工感染实验鱼的肌肉、脾脏等组织中可重新分离到嗜冷黄杆菌。组织病理变化显示,病鱼肝细胞肿胀,空泡变性,部分肝细胞溶解坏死,细胞核溶解消失;脾脏充血、出血,淋巴细胞减少,红细胞和含铁血黄素增多;肌纤维间隙增宽、断裂、弯曲不齐,部分肌细胞肌浆溶解呈蜂窝状。CH06和CH07株对10种抗菌药物的耐药谱略有不同,均对氨苄西林和甲氧苄啶-磺胺甲噁唑敏感;CH06株对恩诺沙星、氟苯尼考等耐药,而CH07株对恩诺沙星和氟苯尼考中度敏感。本研究首次报道了我国虹鳟源嗜冷黄杆菌的分离鉴定及生物学特性,以期为虹鳟细菌性冷水病的防控提供科学依据。     

不同发育阶段的四指马鲅消化道组织学比较研究

采用常规石蜡切片和H-E 染色技术研究不同发育阶段的四指马鲅( Eleutheronema tetradactylum) 消化道的结构特征。结果显示: 1) 35 日龄与43 日龄稚鱼体态相似, 体表被覆鳞片, 背鳍、胸鳍、臀鳍、尾鳍发育健全, 前者全长( 12.850 1.280) mm, 后者( 15.600 1.600) mm。65 日龄幼鱼消化道整体依次分为食道、胃部和肠道, 胃部又分贲门部、幽门部、胃体部和盲囊部, 在幽门部连接肠道处分布4 束短小分散成菜花状的幽门盲囊; 肠道系数为0.630 0.002, 分前、中、后肠。成鱼食道、胃部、肠道直径增大; 肠道系数为0. 300 0.010。2) 两阶段的稚鱼消化道组织结构发育基本与幼鱼、成鱼相似, 消化管壁由内至外依次为黏膜层、黏膜下层、肌肉层和外膜。消化管内黏膜皱褶、胃腺、黏膜上皮杯状细胞的数量在稚鱼、幼鱼、成鱼之间呈递增趋势, 体现结构和功能逐渐发育完善的规律。     

四指马鲅卵巢发育和卵子发生的组织学观察

为了解四指马鲅（Eleutheronema tetradactylum）的卵巢发育和卵子发生过程，本研究运用组织切片HE染色方法对其卵巢发育的组织结构变化和卵子发生过程中各时相卵母细胞的结构进行观察。结果显示，四指马鲅卵巢为被膜型卵巢，紧贴中肾腹面，两支卵巢前端分离于后端融合，呈"Y"字形。卵子发生过程分可分为5个时相，在II时相中期卵母细胞周围开始出现滤泡细胞，II时相晚期形成单层滤泡细胞，直至III时相中期滤泡细胞层外形成鞘膜细胞层；卵黄核在II时相中期开始出现至III时相早期消失；卵黄泡在III时相早期的细胞核附近及胞质边缘出现；卵黄颗粒在III时相晚期开始出现于卵黄泡之间，并在IV时相早期填充卵黄泡，至IV时相晚期形成卵黄小板。卵巢发育过程分为6个时期，每个时期都存在不同时相的卵母细胞；V期卵巢的卵径呈双峰分布，分别在50.00~100.00 μm和300.00~350.00 μm区间出现峰值。四指马鲅卵巢发育模式为非同步发育-分批产卵类型。     

军曹鱼消化系统的形态及组织学研究

利用活体解剖和光镜技术对军曹鱼Rachycentron canadum消化系统进行形态及组织学研究。结果显示,军曹鱼消化道由口咽腔、食道、胃、幽门盲囊、前肠、中肠和后肠几部分组成。口咽腔大,粘膜上皮为复层鳞状上皮,内含少量杯状细胞。食道很短,但上皮包括复层鳞状上皮区和单层柱状上皮区。胃膨大,呈Y形,在贵门和幽门部可观测到杯状细胞,胃体粘膜层下有大量的胃腺细胞。幽门盲囊发达,肠短但分3部分,前肠、中肠和后肠,肠道系数为0．43,肠道由前向后杯状细胞和粘膜皱褶不断减少。消化腺包括肝脏和胰腺,肝小叶分界不明显,肝细胞内脂肪含量丰富,胰腺属于散在性的,其外分泌部有许多腺泡组成,胰岛分散于外分泌部间。     

麻黄鸡禽白血病的诊断及病理组织学观察

通过对广东某麻黄种鸡场抽取的130份样品进行禽白血病抗原ELISA检测和病理组织学观察,结果显示:该鸡场感染禽白血病的阳性率为10%;从病理组织切片中观察到肝脏、脾脏、胰腺、大脑、肺脏和气管等器官组织中有数量不等的髓细胞样瘤细胞呈局灶性或散在增生。根据ELISA检测和病理组织学观察的结果,说明该麻黄祖代鸡群感染了禽白血病。     

稚鳖的性别分化及组织学研究

解剖出生１５～４５ｄ的稚鳖,所有个体的性腺呈细线状,肉眼分辨不出雌雄。组织学研究发现,雌雄个体的性别都已明显分化。雄性精巢的外壁由两层膜构成,其内部由许多隔膜分隔成许多精细管,精细管里充满原始的精母细胞;雌性卵巢有卵巢腔和输卵管,由结缔组织、毛细血管和生殖上皮形成卵巢膜和基质,并把卵巢分隔成一个个小室,原始的卵泡就在这些小室中生长、发育。     

Changes in fertility of rainbow trout eggs retained in coelom

ABSTRACT:    Effects of prolonged retention time of ovulated eggs in the parental coelom on fertilization success were studied in rainbow trout Oncorhynchus mykiss using cryopreserved sperm with a uniform fertilizing ability. Proportions of successful fertilization, eyed eggs and hatched alevins were examined at different time periods up to a retention time of 14 days beyond the ordinary stripping time, and were compared with eggs incubated in artificial coelomic fluids (ACF). Eggs that were retained longer in the coelom showed gradual decreases in all the proportions of successful fertilization, eyed eggs and hatched alevins. The progress of cleavage after fertilization slowed with prolonged retention times. Eggs incubated in ACF lost their fertilizing ability much sooner than those retained in the coelom. The hatching rate of eggs retained for 2 weeks in coelom was 36%, while it was 1% in those eggs incubated for 4 days in ACF. Thus, eggs retained in the coelom showed higher fertilization success than those incubated in ACF.     

Evidence of concealment behavior by adult rainbow trout and brook trout in winter

Abstract – There has been little investigation of the winter ecology of adult trout during winter, especially in regard to concealment behavior. We compared day vs night underwater counts of adult rainbow trout and brook trout from four streams. At water temperatures between 1°C and 9°C, daytime counts accounted for 44% and 16% of nighttime snorkeling counts for rainbow trout and brook trout adults, respectively. As winter progressed, nighttime counts declined more so for brook trout than rainbow trout, but the decline was not significant for either species. Nocturnalism of both species was higher in streams with colder water temperatures. We observed few fish within concealment structure; however, by electrofishing concealment habitat during the day, we captured 10 times more adult trout than we counted immediately beforehand by snorkeling. Adult trout were concealed in cobble-boulder substrate and woody debris during the day. ^Note     

Growth,osmoregulation and energy budget of rainbow and steelhead trout under different salinity acclimation methods and the best transition size of steelhead trout

In order to meet the demand of salmon market, Chinese scientists and entrepreneurs are working on salmon mariculture far offshore in the Yellow Sea, China. Rainbow, steelhead trout and Atlantic salmon were selected as the main culture species. The aims of the present study were as follows: (a) investigate the effect of the salinity acclimation method on the growth, osmoregulation and energy budget in two forms of Oncorhynchus mykiss, rainbow and steelhead trout and (b) explore the optimal size of steelhead trout for the seawater entry. In trial I, rainbow (mean = 99.44 g) and steelhead trout (mean = 99.01 g) were reared for 40 days after undergoing salinity acclimation at three rates: an abrupt increase in salinity from 0 to 30 g/L (T30); an abrupt increase in salinity to 14 g/L, followed by a daily increase of 2 g/L (T2) or 6 g/L (T6) until reaching 30 g/L; and no salinity exposure (control treatment) (T0). In trial II, steelhead trout with body weights of approximately 100 and 400 g were cultured for 60 days with two treatments, T0 and T2, and the specific growth rate (SGR) was calculated every 10 days. In trial I, in both kinds of fish, the optimal growth performance, survival rate, osmoregulation and energy budget were observed in the T0 treatment, followed by the T2 treatment. These results indicate that O. mykiss with a body weight of approximately 100 g can adapt to sea water with a gradual transition (T2), but they are still not suitable for the seawater entry because of low growth. Based on the recorded SGRs in trial II, our formulated regression formula revealed that approximately 200 g is the optimal size of steelhead trout for the transition to sea water.     

Temporal segregation in spawning between native Yellowstone cutthroat trout and introduced rainbow trout

Hybridisation with introduced taxa poses a threat to native fish populations. Mechanisms of reproductive isolation can limit or prevent hybridisation between closely related species. Understanding how these mechanisms interact between the same species across geographically distinct occurrences of secondary contact, and how regional factors influence them, can inform our understanding of hybridisation as a threat and management actions to mitigate this threat. We used data collected on adult fish migration timing and approximate emergence timing of subsequent juvenile fish paired with genomic data to assess whether temporal isolation in the timing of spawning exists between Yellowstone cutthroat trout, rainbow trout and hybrids in the North Fork Shoshone River drainage in northwest Wyoming. We found evidence that Yellowstone cutthroat trout spawn, on average, two to four weeks later than rainbow trout and hybrids and two environmental covariates related to water temperature and discharge were associated with differences in spawning migration timing. Despite statistical support for Yellowstone cutthroat trout spawning later, disproportionately high numbers of rainbow trout and hybrids, paired with extended spawning seasons, lead to substantial overlap between all genotypes. Our results provide further evidence of temporal segregation in the timing of spawning as a mechanism of reproductive isolation between closely related species, but substantial spawning overlap suggests temporal segregation alone will not be enough to curtail hybridisation in conservation populations.     

虹鳟IgM重链恒定区的表达及兔抗血清的制备

为了能够成功表达虹鳟IgM,本研究利用生物信息学软件对虹鳟IgM的亲水性及抗原性进行了分析,根据GenBank收录的虹鳟IgM重链恒定区,参照生物信息学分析结果,设计用于扩增截短的IgM基因的引物,以虹鳟头肾RNA提取物为模板,利用RT-PCR方法扩增虹鳟截短的IgM重链恒定区部分基因片段,连接原核表达载体pET-27b,利用大肠杆菌Rosetta进行表达。SDS-PAGE及HPLC结果显示,纯化后截短的IgM大小约为47.7 ku,且纯度达到90%。利用其制备兔抗血清后ELISA分析结果显示,所制备的兔抗血清与本研究所表达的截短IgM蛋白的反应效价为1∶40 000,与虹鳟血清提取的全长IgM反应效价为1∶20 000并且呈现出抗原计量依赖性。研究表明,重组IgM蛋白与虹鳟血清中的天然IgM重链恒定区具有近似的结构,利用其所制备的兔抗血清能够与虹鳟鱼体中的天然IgM发生特异性反应。     

Experimental pathogenicity in rainbow trout, Oncorhynchus mykiss (Walbaum), of two distinct morphotypes of long-spined Saprolegnia isolates obtained from wild brown trout, Salmo trutta L., and river water

Juvenile rainbow trout, Oncorhynchus mykiss (Walbaum), were experimentally infected to investigate the pathogenicity of 20 isolates of two morphotypes of long-haired Saprolegnia obtained from wild brown trout, Salmo trutta L., and river water in Spain. The trout were exposed to 2 × 10⁵ and 3 × 10⁵ L^–1 zoospores. Saprolegnia infection could not occur without 'ami-momi' treatment. Pathogenicity varied greatly among isolates as mortality ranged from 0 to 100% of the fish. There was a statistically significant difference ( P  < 0.001) between the mortality caused by morphotype I isolates and that produced by those of morphotype II. The most pathogenic isolates usually belonged to morphotype II, consisting of isolates which had secondary cysts with bundles of hooked hairs which were shorter and less numerous than those of morphotype I; the morphotype I isolates usually had low pathogenicity. Lesions were most frequently found on the fins. Cultures detected the presence of Saprolegnia in internal organs. Histopathology of the intestine suggests that Saprolegnia may reach this and other organs via the blood stream from surface lesions.     

A study of gross,histological and blood biochemical changes in rainbow trout,Oncorhynchus mykiss (Walbaum), with rainbow trout gastroenteritis (RTGE)

The mechanisms behind the pathogenesis of rainbow trout gastroenteritis (RTGE) are still unknown. This study examined the macroscopic and microscopic changes in trout with RTGE (RTGE+), as well as the blood chemistry. A total of 464 rainbow trout were sampled from 11 sites in the UK, comprising 152 RTGE+ fish and 330 random, apparently healthy fish. A case definition for RTGE was assessed by the analysis of its agreement with three laboratory tests: histopathology, packed cell volume and kidney bacteriology. Cluster analysis indicated the presence of three distinct presentations within the population of RTGE+ fish. Cluster A included gross signs associated with moribund RTGE+ fish, and clusters B and C identified gross signs consistent with concurrent diseases, notably furunculosis, enteric redmouth and proliferative kidney disease. The information gained was used to select RTGE+ fish without concurrent disease for the analysis of RTGE pathogenesis with blood biochemistry. This analysis revealed a severe osmotic imbalance and a reduced albumin/globulin ratio as indicatives of selective loss of albumin. These findings are compatible with a protein losing enteropathy.     

Effects of various diet formulations (experimental and commercial) on the morphology of the liver and intestine of rainbow trout (Oncorhynchus mykiss) juveniles

The aim of the present study was to evaluate the effects of three formulated diets: wheat gluten protein‐based diets supplemented with Lys–Gly dipeptide (PP) or free lisine and glycine (AA), and a control diet without lysine supplementation (CON), two commercial starters: Aller Futura (AF), Aglo Norse (AN) and frozen Chironomidae larvae (CH) for on the growth and digestive system development of rainbow trout. The experiment lasted 28 days and survival at the end of the experiment was the highest in the group fed AF (87.9±4.2%) and the lowest in the group fed the CON diet (74.0±4.3%). Fish fed the CON diet showed a significantly lower crude protein content, and the AA group had a significantly higher lipid level (P≤0.05) in body carcass compared with the other groups. The highest density of intestinal goblet cells (P≤0.05) was observed in fish fed the CON diet, while the lowest density was seen in the distal intestine in the AF group. The hepatocyte cytoplasm of all fish stored more glycogen than lipids. These results indicate that wheat gluten protein‐based diets supplemented with dipeptide Lys–Gly (PP) or free lysine and glycine (AA) supported growth, protein efficiency rate and intestine epithelium homeostasis. On the other hand, fish fed CON (lysine‐deficient diet) showed lower survival and growth rate, and disturbances of intestine epithelium homeostasis, probably as a result of lysine deficiency.