The effect of dietary folic acid on biochemical parameters and gene expression of three heat shock proteins (HSPs) of blunt snout bream (<Emphasis Type="Italic">Megalobrama amblycephala)</Emphasis> fingerling under acute high temperature stress

The effects of dietary folic acid on biochemical parameters and gene expression of three heat shock proteins (HSPs) of blunt snout bream (Megalobrama amblycephala) fingerling under acute high temperature stress. Six dietary folic acid groups (0.0, 0.5, 1.0, 2.0, 5.0, and 10.0) mg/kg diets were designed and assigned into 18 tanks in three replicates each (300 l/tank) and were administered for 10 weeks in a re-circulated water system. The fingerlings with an initial weight of 27.0 ± 0.03 g were fed with their respective diets four times daily. At the end of the experiment, samples were collected before challenge, 0, 24, 72 h, and 7 days. Serum total protein (TP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), cortisol, glucose, complement C3 (C3), complement C4 (C4, immunoglobulin M (IgM) hepatic superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), and the expression of heat shock protein 60 (HSP60), 70 (HSP70), and 90 (HSP90) were studied. The results showed that fish fed with dietary folic acid between 1.0, 2.0, and 5.0 mg/kg significantly (P < 0.05) increased serum TP, C3, C4 hepatic SOD, CAT, and the expression of HSP60, HSP70, and HSP90 before and after temperature challenge of 32 °C. Also, serum ALP, cortisol, glucose, and hepatic MDA were significantly (P < 0.05) reduced by supplementation of dietary folic acid level 1.0, 2.0, and 5.0 mg/kg before and after the same temperature challenge of 32 °C. Before stress, 0, 24, 72 h, and 7 days significantly (P < 0.05) affects serum biochemical parameters, immune and antioxidant capacities, and expression level of three HSPs. Furthermore, there was no statistical evidence to show that dietary folic acid inclusion level and temperature duration have significant interactive effect on serum biochemical parameters, antioxidant parameters, and gene expression level (P > 0.05) of the three HSPs. However, there were statistical significant interactive effect between dietary folic acid inclusion level and temperature duration on serum C3 and C4 (P < 0.05) except IgM (P > 0.05). The present results indicate that supplementation of basal diet from 1.0 mg/kg; 2.0 and 5.0 mg/kg can enhance acute high temperature resistance ability in M. amblycephala fingerling to some degree and improve physiological response, immune and antioxidant capacities, and expression level of three HSPs.     

Comparison of Conventional Washing Processing and pH Shift Processing on Gelation Characteristics of Bighead Carp (Aristichthys nobilis) Muscle Proteins

In this study, gelation characteristics of bighead carp (Aristichthys nobilis) protein isolates prepared by pH shift processing were investigated as compared with those of surimi prepared by conventional wash processing. Results showed that the gel from alkali-aided protein isolate exhibited a higher breaking force than that from conventional washed surimi (p < 0.05). However, the gel from conventional washed surimi had higher whiteness (p < 0.05) and lower expressible moisture (p < 0.05). The rheological study showed that protein isolates exhibited different storage modulus and loss tangent patterns from conventional washed surimi during heating. Acid-aided processing could lead to higher denaturation and dissociation for fish muscle myosin and a coarser gel network from the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) pattern and the scanning electron microscopy (SEM). Overall, there is a potential to apply pH shift processing, especially alkali-aided processing, to extract more proteins from bighead carp muscle for the production of surimi-based food.     

锦江鱼类资源现状与物种多样性分析

2013年8月—2016年8月对沅江水系支流锦江的鱼类资源现状进行了调查。通过向渔民购买、现场捕捞等方式,共调查野生鱼类43种,隶属于5目11科39属,其中鲤科鱼类25种,占总种类数的58.1%。调查到新记录种5种,分别为大鳞副泥鳅(Paramisgurnus dabryanus)、吻(Rhinogobio typus)、洞庭华鲮(Sinilabeo tungting)、云南盘(Discogobio yunnanensis)和大眼鳜(Siniperca kneri)。干、支流均以底层鱼类为主,种类数分别占各自总数的55.8%和71.4%。对比历史资料,G-F多样性指数呈下降趋势。对干流渔获物种类的周年统计发现,出现频率在50%以上的鱼类有8种,其中泥鳅(Misgurnus anguillicaudatus)、鲤(Cyprinus carpio)、鲫(Carassius auratus)和鲇(Silurus asotus)的出现频率均达到97.3%。     

Characterization of <Emphasis Type="Italic">kiss2</Emphasis> and <Emphasis Type="Italic">kissr2</Emphasis> genes and the regulation of kisspeptin on the HPG axis in <Emphasis Type="Italic">Cynoglossus semilaevis</Emphasis>

Reproduction allows organisms to produce offspring. Animals shift from immature juveniles into mature adults and become capable of sexual reproduction during puberty, which culminates in the first spermiation and sperm hydration or ovulation. Reproduction is closely related to the precise control of the hypothalamic–pituitary–gonadal (HPG) axis. Kisspeptin peptides are considered as the important regulator of HPG axis in mammalian. However, the current understanding of kisspeptin in flatfish is not comprehensive. In this study, we cloned and analyzed the kiss2 and kissr2 genes in Cynoglossus semilaevis. Interesting alternative splicing in the 5′-untranslated regions (UTR) of the Cskissr2 gene was found. The expression profiles of Cskiss2 and Cskissr2 showed relative high messenger RNA (mRNA) levels at the late gastrula stage during embryonic development, at total length = 40 mm during early gonadal differentiation, and in the brains and gonads of all investigated tissues. These results suggested that the kisspeptin system participated in embryogenesis and in the regulation of gonadal differentiation and development. Considering that the control and regulatory mechanisms of kisspeptin in the central reproductive axis are still unclear, we documented that the intramuscular injection of kisspeptin caused different sGnRH and cGnRH mRNA levels in a dose- and tissue-dependent manner. The mRNA expressions of FSH and LH were stimulated in the ovary and were inhibited in the testis under the kisspeptin treatments. These results provided foundations for understanding the roles of kisspeptin in the neuroendocrine system in fish. The manipulation of the kisspeptin system may provide new opportunities to control the gonadal development and even reproduction in fish.     

不同施氮水平下缓释氮肥配施对机插稻氮素利用特征及产量的影响

【目的】在高施氮水平和常规施氮水平下,研究缓释氮肥配施对机插稻氮素利用和产量的影响及其生理机制,为机插稻的氮肥高效利用提供依据。【方法】以中迟熟杂交籼稻川谷优7329为材料,采用二因素裂区设计,设置不同的施氮水平及5种缓释氮肥与常规氮肥(尿素)配施处理,并设置不施氮处理,分析不同施氮水平下缓释氮肥与常规氮肥配施对机插稻氮素利用和产量的影响及其生理机制,并探讨氮素利用和产量与生理响应之间的关系。【结果】结果表明,施氮水平、缓释氮肥与常规氮肥配施对机插稻氮素利用特征及产量均存在显著或极显著的影响,机插稻拔节、抽穗及结实期对氮素的吸收利用以及结实期茎鞘的氮素转运量与干物质量、氮素积累量、氮肥表观利用率、每穗实粒数及最终产量显著或极显著正相关(r=0.38*~0.69**)。与常规氮肥运筹相比,缓释氮肥与常规氮肥配施、全缓释氮肥施用处理的机插稻成熟期干物质积累量、氮素积累总量、光合势、叶面积指数、氮肥表观利用率及穗部氮素增加量均显著提高,进而促进增产。【结论】据产量及氮素利用效率表现,机插稻产量在180 kg/hm2施氮水平下较高,且在缓释氮肥与常规氮肥配施比例为7∶3一次性施用时产量最高,较其他氮肥运筹产量高0.84%~26.59%,氮肥表观利用率高0.28%~47.02%,为最优氮肥运筹模式。但不同施氮水平下,随着缓释氮肥配施比例的降低,群体叶面积指数、光合势及物质积累等指标也降低,且在施氮处理为全常规氮肥时最低,均不利于机插稻氮肥利用效率及最终产量的提高。     

Effect of N-acetyl cysteine and glycine supplementation on growth performance,glutathione synthesis,and antioxidative ability of grass carp, <Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis>

An 8-week feeding trial was conducted to evaluate the effect of N-acetyl cysteine (NAC) and glycine supplementation on growth performance, glutathione (GSH) synthesis, and antioxidative ability of grass carp, Ctenopharyngodon idella. Four practical diets were formulated: control, control + 0.2% NAC, control + 0.5% glycine, and control + 0.2% NAC + 0.5% glycine. Each diet was randomly assigned to quadruplicate groups of 30 fish (approximately 8.8 g). Weight gain and specific growth rate were significantly increased with the supplementation of NAC. Supplementation of NAC plus glycine significantly increased the feed efficiency. Glutathione peroxidase (GPx) and γ-glutamine cysteine synthase (γ-GCS) in plasma were significantly increased with the supplementation of NAC plus glycine. GSH in plasma increased and malondialdehyde (MDA) decreased in fish fed diets supplemented with NAC. Respiratory burst, superoxide dismutase (SOD), and catalase (CAT) activity were not affected by NAC or glycine. These results clearly indicated that NAC improved the growth performance and restored GSH of grass carp, supplemented NAC together with glycine enhanced GSH synthesis, and improved the antioxidative ability of grass carp.     

Invasive Chinese pond mussel Sinanodonta woodiana threatens native mussel reproduction by inducing cross‐resistance of host fish

1. The effects of invasive alien species (IAS) on host–affiliate relationships are often subtle and remain unnoticed or insufficiently quantified. The global decline of freshwater unionid mussel species has been attributed to many causes, but little is known about the interactions of IAS, with their complex life cycle, which includes an obligatory parasitic stage (the glochidium) that develops on fishes.
2. The capacity of a European freshwater mussel, Anodonta anatina, to develop on its widespread fish host, Squalius cephalus was tested experimentally, after previous infestations by the IAS, Sinanodonta (Anodonta) woodiana. The initial attachment of glochidia, the length of the parasitic period, and the metamorphosis success rate of A. anatina glochidia were compared among treatments of different priming infestation intensities.
3. The metamorphosis success rate of the native A. anatina glochidia was strongly reduced (Wilcoxon Signed‐Rank Test, P < 0.001) and declined by 42.1 and 45.4% on fish hosts that were previously exposed to S. woodiana by single and multiple priming infestations, respectively, in comparison with the control group. Such cross‐resistance is expected to decrease significantly the quality of the host resources available to native mussels.
4. This study provides the first evidence of the host‐mediated adverse impact of invasive S. woodiana on native mussel species. These results also highlight the importance of potential competition for hosts between threatened groups of affiliate species and their invasive counterparts, which should be reflected in conservation strategies.

     

Plasma levels of immune factors and sex steroids in the male seahorse <Emphasis Type="Italic">Hippocampus erectus</Emphasis> during a breeding cycle

To better understand the endocrine- and immune-response pattern during reproduction in a fish species having parental care behaviors and also to accumulate the endocrine- and immune-related data for future explanations of the low reproductive efficiency in seahorse species, the variations of immune factors and sex steroids in the plasma of the male lined seahorse Hippocampus erectus at different breeding stages, i.e., pre-pregnancy, pregnancy (early, middle, and late periods), and post-pregnancy, were investigated in the present study. The immune factors included monocytes/leucocytes (M/L), leucocyte phagocytic rate (LPR), immunoglobulin M (Ig M), interleukin-2 (IL-2), interferon-α (IFN-α), and lysozyme (LZM). The sex steroids included testosterone (T), 11-ketotestosterone (11-KT), 11β-hydroxytestosterone (11β-OHT), 17α-methyltestosterone (17α-MT), 17β-estradiol (E2), and 17α-hydroxy-20β-dihydroprogesterone (17α-20β-P). Moreover, the immune metabolic activity of epithelium cells in the brood pouch at different breeding stages was also analyzed through ultrastructural observations of the abundance of cytoplasmic granules, mitochondria, endoplasmic reticulum, lysosomes, and exocytosis. The results show that a higher immune level was observed during pregnancy, particularly in the early and middle periods, and a lower immune level was noted during pre-pregnancy. Correspondingly, the epithelium cells in the brood pouch also showed a stronger immune metabolic activity during pregnancy and weaker activity during pre-pregnancy. Four sex steroids of T, 11β-OHT, 17α-MT, and E2 were higher during pre-pregnancy and lower during post-pregnancy, whereas 11-KT and 17α-20β-P, which were positively correlated with part immune factors, were higher during pregnancy. No negative correlations between sex steroids and immune factors were observed. In conclusion, the higher immune competence during pregnancy may indicate that parental care could improve immunity, which may be the major factor for no immunosuppressive effect of sex steroids during reproduction in the seahorse H. erectus, unlike noncaregiving fishes in which inhibitions of sex steroids on immunity are frequently observed. Moreover, higher 11-KT and 17α-20β-P during pregnancy than during pre-pregnancy and post-pregnancy may suggest that these two steroids are also involved in parental care regulation.     

白斑综合证病毒胶体金免疫层析试剂板的研制

将胶体金免疫层析技术应用于对虾白斑综合征的诊断,研制了一种检测对虾中白斑综合征病毒的胶体金免疫层析试剂板.将胶体金标记的抗白斑综合征病毒单克隆抗体包被在金标垫上,将抗白斑综合征病毒多克隆抗体和羊抗鼠IgG分别包被在硝酸纤维素膜的检测线和质控线上.通过双抗体夹心法检测样品中自斑综合征病毒含量.结果表明,该试剂板对白斑综合征病毒的最低检出限为1.0 mg/kg,特异性好,稳定性高,整个检测所需时间为8～10 min,适合现场快速检测.     

急性操作胁迫对四指马鲅幼鱼肝脏组织结构和氧化应激的影响

为探究急性操作胁迫对四指马鲅(Eleutheronema tetradactylum)幼鱼肝脏组织结构以及氧化应激的影响,该研究在离水操作胁迫后不同时间取样,观察肝脏显微结构的变化,并检测相关抗氧化酶活性。结果显示,随着离水胁迫时间的延长,肝脏组织损伤程度呈现先上升后下降的趋势,胁迫第24小时肝脏基本结构与对照组相似。超氧化物歧化酶(SOD)和过氧化氢酶(CAT)有相同的变化趋势,胁迫后第2小时显著增加(P0.05)并达到最高值,然后开始下降,最后其值显著低于对照组(P0.05);丙二醛(MDA)含量在胁迫第2~第12小时呈现先升高后下降的变化趋势,第12小时后又有所上升;还原型谷胱甘肽(GSH)含量胁迫2 h没有显著变化(P0.05),之后呈先下降后上升的趋势,但其含量仍显著低于对照组(P0.05);总抗氧化能力(T-AOC)胁迫第2小时显著上升(P0.05),之后下降,实验结束时显著低于对照组(P0.05)。结果表明离水操作胁迫对四指马鲅幼鱼肝脏组织结构以及抗氧化酶产生一定的影响,但随着胁迫时间的延长,其组织结构以及抗氧化能力都有一定的恢复趋势。