齐口裂腹鱼血细胞的发生

为了探讨齐口裂腹鱼(Schizothorax prenanti)血细胞发生发育的模式,采用Wright’s、碘酸雪夫氏(PAS)和苏丹黑B(SBB)染色方法对齐口裂腹鱼的头肾、中肾、脾和肝组织印片进行染色,了解其发生的具体组织和发育的一般过程。结果表明:齐口裂腹鱼血细胞可分为红细胞系、淋巴细胞系、单核细胞系、粒细胞系和其他细胞,且这些细胞系的发育均经过3个阶段,即原始阶段、幼稚阶段、成熟阶段。通过观察和统计不同阶段各种血细胞的形态、大小、比例及染色特征,发现原始阶段的血细胞体积较大,其中原始单核细胞最大,大小为(18.90±1.59)μm×(16.32±0.70)μm,在4种组织中原始阶段的红细胞和单核细胞在头肾中所占比例最大,分别为0.64%和0.59%,原粒细胞和原淋巴细胞分别在中肾和脾中比例最大,在肝中少量存在,此外在各组织印片中均发现血栓细胞的存在,在肝中发现巨噬细胞。因此头肾、中肾和脾是齐口裂腹鱼的主要造血组织,少量细胞在肝中产生。粒细胞发育过程中,除原始粒细胞PAS染色阴性外,其他阶段均呈阳性,且阳性随不断成熟逐渐增强;单核细胞从原单核细胞到成熟的单核细胞,PAS阳性逐渐增强;而SBB染色发现,粒细胞、淋巴细胞及单核细胞均呈阳性,未成熟的细胞染色程度不一致,成熟的细胞阳性染色最为强烈。在红细胞系发育过程中经历了细胞由大变小再变大的过程,而粒细胞和淋巴细胞发育过程仅出现由大变小的过程,在红细胞的发育过程中还出现了染色质固缩和血红蛋白增加。     

黄芪多糖对齐口裂腹鱼生长、体组成和免疫指标的影响

试验研究黄芪多糖对齐口裂腹鱼生长性能、体组成及免疫指标的影响。以450尾健康的齐口裂腹鱼[体重(6.98±0.43)g;体长(9.11±0.25)cm]为试验对象,随机分为5组(C1、C2、C3、C4、C5),每组3个重复,每重复30尾试验鱼。C1、C2、C3、C4、C5组分别投喂在等氮等能(蛋白质含量38.29%,能量15.73 mJ/kg)的基础料中分别添加0、0.02、0.04、0.06、0.08%的黄芪多糖制成5种试验饲料,养殖齐口裂腹鱼50d。结果表明:饲料中未添加黄芪多糖组的增重率(WGR)、特定生长率(SGR)、饲料蛋白效率(PER)均显著低于黄芪多糖添加组(P<0.05),而饵料系数(FCR)则显著高于黄芪多糖添加组(P<0.05)。当黄芪多糖添加水平为0.04%时,试验鱼的WGR、SGR、PER均达到最大(分别为110.31%、1.86%/d和182.07%),FCR最低(1.44),与其他各组差异显著(P<0.05);以WGR、SGR、PER、FCR为指标,利用直线和抛物线回归分析表明,齐口裂腹鱼生长性能最佳时黄芪多糖添加水平为0.045%—0.074%;对齐口裂腹鱼机体组成分析表明,黄芪多糖对鱼体粗灰分和水分影响不显著(P>0.05),黄芪多糖添加水平为0.06%时机体粗蛋白最高,但与黄芪多糖添加水平为0.04%时无明显差异(P>0.05);粗脂肪含量在黄芪多糖添加水平为0.04%时最高,与其他各组差异显著(P<0.05);未添加黄芪多糖组血清免疫酶活性显著低于黄芪多糖添加组,齐口裂腹鱼血清免疫酶活性在一定范围内随黄芪多糖的增加而增强。黄芪多糖添加水平为0.04%时,碱性磷酸酶(ALP)活性最高;酸性磷酸酶(ACP)活性在黄芪多糖添加水平0.06%时最大;黄芪多糖添加水平应在0.06%—0.08%时,溶菌酶(LSZ)、超氧化歧化酶(SOD活性趋于稳定。这说明黄芪多对齐口裂腹鱼的生长和免疫力有明显的促进作用。综合考虑齐口裂腹鱼生长性能和免疫能力最佳时黄芪多糖添加水平为0.04%—0.074%。     

四川裂腹鱼脑型芳香化酶Cyp19b基因组织分布及温度对其表达水平的影响

为研究脑型芳香化酶基因Cyp19b在四川裂腹鱼(Schizothorax kozlovi)早期性别分化中的作用,采用RACE方法从四川裂腹鱼脑中扩增得到该基因c DNA全长序列,并应用荧光定量RT-PCR技术测定该基因m RNA的相对表达量,探讨该基因在不同规格四川裂腹鱼鳃、脑、心、肝、脾、肾、肌肉、精巢、卵巢组织中的表达差异,以及温度对其早期仔鱼阶段该基因表达的影响。四川裂腹鱼Cyp19b基因的c DNA全长序列共3 021 bp,共编码507个氨基酸,属于脑型芳香化酶基因;四川裂腹鱼Cyp19b基因编码的氨基酸序列与其他鱼类脑型芳香化酶基因编码的氨基酸序列同源性可达70%以上,而与性腺型芳香化酶基因编码的氨基酸序列同源性较低,为64%左右;四川裂腹鱼Cyp19b基因仅在脑组织中表达,具有比较高的组织特异性,且随着个体的增长,逐渐呈现显著的雌雄差异;对不同温度处理12日龄仔鱼6 d后,低温(10℃和14℃)能显著促进Cyp19b基因表达量的升高,但高温(26℃)却对其表达量无显著影响。由此推测,脑型芳香化酶基因Cyp19b可能在低温导致四川裂腹鱼雌性化过程中发挥着重要作用。     

Genetic diversity and evolutionary history of the Schizothorax species complex in the Lancang River (upper Mekong)

The genus Schizothorax (Cyprinidae), one of the most diverse genera of ichthyofauna of the Qinghai‐Tibetan Plateau (QTP), is a good candidate for investigating patterns of genetic variation and evolutionary mechanisms. In this study, sequences from the mitochondrial control region, the cytochrome b gene, and two nuclear genes were used to re‐examine the genetic diversity and investigate the evolutionary history of the Schizothorax species complex inhabiting the Lancang River. Three maternal clades were detected in the Schizothorax species complex, but frequent nuclear allele sharing also occurred among the three maternal clades. A discrepancy between topologies of mitochondrial and nuclear loci might result from introgression or/and incomplete lineage sorting. The divergence of the clades of the Schizothorax species complex was closely related to the Late Pliocene and Early Pleistocene orogenesis of the QTP and Southwest Mountains of China. Demographic analyses indicated that the species complex subsequently persisted in situ with stable populations during Pleistocene glacial cycling, which suggested that Pleistocene climate changes did not exert a remarkable influence on the species complex. Our study provides a comprehensive analysis of the genetic diversity and evolutionary history of the Schizothorax species complex in the Lancang River.     

饲料蛋白质和能量水平对齐口裂腹鱼生长、体组成和蛋白质利用的影响

齐口裂腹鱼（Schizothorax prenanti Tchang）隶属鲤科、裂腹鱼亚科,俗称“雅鱼”,为长江上游重要经济鱼类,也是川西高原特有的冷水性鱼类。其肉质细嫩,味道鲜美,是名贵野生经济鱼类。近年来由于过度捕捞、环境污染等因素的影响,齐口裂腹鱼种群数量不断下降,分布区域日趋缩小,而随着人们生活水平的不断提高,对齐口裂腹鱼等名优水产品的需求却在不断增加。为保护和持续利用这一野生动物资源,     

基于mtDNA Cyt b序列分析齐口裂腹鱼群体遗传多样性

研究测定了长江上游4个齐口裂腹鱼(Schizothorax prenanti)野生群体(重庆巫溪、重庆城口、四川雅安、四川阿坝)共104个个体的线粒体Cyt b基因部分序列, 以探讨齐口裂腹鱼野生群体的遗传多样性和遗传结构。结果表明: 在104个个体Cyt b序列中共检测到43个多态性位点, 25个单倍型。4个齐口裂腹鱼群体的单倍型多样性介于0.704—0.884, 核苷酸多样性介于0.007—0.012。群体间Kimura双参数遗传距离介于0.008—0.017, 其中四川雅安群体与四川阿坝群体间遗传距离最近, 基因交流频繁。重庆城口群体与四川雅安群体间遗传距离最远, 基因交流受阻。AMOVA分析表明, 齐口裂腹鱼的遗传分化主要来自群体内部, 且组群间、组群内群体间和群体内存在显著的遗传分化。中性检验得到Tajima’s D和 Fu’s F_s的值不显著, 且歧点分布图呈多峰, 表明长江上游4个齐口裂腹鱼野生群体未经历过种群扩张。研究旨为齐口裂腹鱼野生资源保护提供必要参考意见, 同时为齐口裂腹鱼种质资源合理开发和利用提供理论依据。     

齐口裂腹鱼脑内5-羟色胺免疫组织化学的定位观察

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齐口裂腹鱼耳石早期生长发育与日轮特征研究

文章研究了在实验室条件下齐口裂腹鱼仔稚鱼耳石早期形态发育与生长特点、第一轮纹出现时间和轮纹沉积规律。结果表明: 在13.5-17.2℃孵化条件下,微耳石和矢耳石在出膜前形成,而星耳石于出膜后第12天出现。在仔稚鱼生长过程中,微耳石由近圆形发育成贻贝形,矢耳石经历近圆形、锲形后发育为箭矢状,星耳石形状由近圆形发育为星芒状。微耳石的前区、背区和腹区及矢耳石的背区和腹区生长呈幂函数关系,而微耳石的后区、矢耳石前区和后区生长以及两对耳石的前后区半径之和与全长均呈线性相关。在(18.50.5)℃和(15.61.1)℃条件下,50%矢耳石样本第一轮纹均在出膜后第 2 天形成(分别为出膜后18h和19h),以后每天形成一轮。微耳石和矢耳石轮纹数均与日龄呈线性相关,方程斜率均与1差异不显著(P0.05),表明两对耳石的轮纹沉积均为日周期性。这些结果为研究齐口裂腹鱼野生种群繁殖期和早期生活史特征等生态学问题提供了重要依据。     

Biogeography and molecular phylogeny of the genus Schizothorax (Teleostei: Cyprinidae) in China inferred from cytochrome b sequences

Aim To test a vicariant speciation hypothesis derived from geological evidence of large‐scale changes in drainage patterns in the late Miocene that affected the drainages in the south‐eastern Tibetan Plateau. Location The Tibetan Plateau and adjacent areas. Methods The cytochrome b DNA sequences of 30 species of the genus Schizothorax from nine different river systems were analysed. These DNA sequences were analysed using parsimony, maximum likelihood and Bayesian methods. The approximately unbiased and Shimodaira–Hasegawa tests were applied to evaluate the statistical significance of the shortest trees relative to alternative hypotheses. Dates of divergences between lineages were estimated using the nonparametric rate smoothing method, and confidence intervals of dates were obtained by parametric bootstrapping. Results The phylogenetic relationships recovered from molecular data were inconsistent with traditional taxonomy, but apparently reflected geographical associations with rivers. Within the genus Schizothorax, we observed a divergence between the lineages from the Irrawaddy–Lhuit and Tsangpo–Parlung rivers, and tentatively dated this vicariant event back to the late Miocene (7.3–6.8 Ma). We also observed approximately simultaneous geographical splits within drainages of the south‐eastern Tibetan Plateau, the Irrawaddy, the Yangtze and the Mekong–Salween rivers in the late Miocene (7.1–6.2 Ma). Main conclusions Our molecular evidence tentatively highlights the importance of palaeoriver connections and the uplift of the Tibetan Plateau in understanding the evolution of the genus Schizothorax. Molecular estimates of divergence times allowed us to date these vicariant scenarios back to the late Miocene, which agrees with geological suggestions for the separation of these drainages caused by tectonic uplift in south‐eastern Tibet. Our results indicated the substantial role of vicariant‐based speciation in shaping the current distribution pattern of the genus Schizothorax.     

Effects of substrate roughening on the swimming performance of Schizothorax wangchiachii (Fang, 1936) in the Heishui River: Implications for vertical slot fishway design

Re-establishing the natural connectivity of rivers using fishways may mitigate the unfavourable effects of dam construction on riverine biodiversity and freshwater fish populations. Knowledge of the swimming performance of target species in specific regions is critical for designing fishways with a high passage efficiency. Substrate roughening with river stones of fishways is considered to improve fish swimming capacity by benefiting from reduced-velocity zones with lower energetic costs. However, the effectiveness of rough substrates in energy metabolism is rarely tested. We investigated the effect of substrate roughening on the swimming capacity, oxygen consumption and behaviour of Schizothorax wangchiachii from the Heishui River in a flume-type swimming respirometer. The results showed that substrate roughening improved critical and burst swimming speed by ~12.9% and ~15.0%, respectively, compared to the smooth substrate. Our results demonstrate that increased reduced-velocity zones, lowered metabolic rate and tail-beat frequency support our hypothesis that lower energetic costs improve fish swimming performance in rough substrate compared to smooth treatment. The traversable flow velocity model predicted that maximum traversable flow velocity and maximum ascent distance were higher over rough compared to smooth substrate fishways. Fishway substrate roughening may be a practical approach to improve fish swimming upstream for demersal riverine fish.