猪GRP78基因克隆、生物信息学分析及组织表达谱研究

本研究旨在克隆猪葡萄糖调节蛋白78（glucose-regulated protein 78，GRP78）基因，并进行生物信息学分析，探讨其在猪不同组织中的表达情况。根据GenBank上公布的猪GRP78基因序列（登录号：XM_001927795.6）设计引物，PCR扩增及测序获得猪GRP78基因CDS序列，使用在线软件分析GRP78的理化性质、跨膜结构、信号肽、疏水性、保守结构域、二级结构和三级结构，并进行同源性比对及系统进化树构建，利用实时荧光定量PCR方法检测猪GRP78基因在各组织中的表达情况。结果显示，猪GRP78基因CDS区长1 965 bp，可编码654个氨基酸。生物信息学分析表明，GRP78理论分子质量为72.3 ku，等电点（pI）为5.06，半衰期为30 h，其水溶液在280 nm处的消光系数为30 495，肽链N端为蛋氨酸（Met），不稳定系数为32.39，属于稳定蛋白。脂肪系数为85.40，总平均疏水指数为-0.496，无跨膜结构，存在信号肽序列，说明该蛋白属于分泌型蛋白；GRP78蛋白只包含1个超家族保守结构域：HSP70结构域。蛋白二级结构分析显示，GRP78蛋白中α-螺旋、延伸链、β-转角和无规则卷曲分别为40.06%、20.49%、8.10%和31.35%。同源性比对结果显示，猪GRP78基因与人（登录号：NM_005347.4）、小鼠（登录号：NM_001163434.1）、大鼠（登录号：NM_013083.2）、山羊（登录号：XM_005687138.3）、牛（登录号：NM_001075148.1）核苷酸序列的同源性分别为93%、91%、90%、95%、95%，氨基酸序列的同源性分别为99%、98%、98%、99%、99%，各个物种之间GRP78基因保守性较高。实时荧光定量PCR结果显示，GRP78基因在心脏、肝脏、脾脏、肺脏、肾脏、小肠、胃、卵巢、输卵管、乳腺、小脑、大脑、垂体等组织中均有表达，在心脏、脾脏、肺脏中相对高表达。本研究结果为今后深入研究GRP78基因的生物学功能提供了基础材料。     

施用锌肥和遮阴互作对花生生长发育、抗病性及产量的影响

采用大田裂区试验，研究了施用锌肥和不同遮阴程度互作对花生生长发育、抗病性及产量的影响。结果表明，与不施锌肥相比，施用锌肥能提高花生不同部位锌含量、增加叶片SPAD值，提高花生叶片中可溶性糖、蛋白质和生长素含量，减少花生病害的发生，平均降低7.1个百分点，花生产量平均增加19.4%。相同施锌水平下，随着遮阴程度的增加，花生不同部位锌的含量和不同生育期叶片SPAD值以及花生叶片中可溶性糖、蛋白质和生长素含量呈增加趋势，花生的发病率比不遮阴对照增加4.8、10.2个百分点，花生产量平均降低16.5%、10.0%。在30%、70%的遮阴条件下，施用锌肥的花生产量比不施锌的分别提高21.1%、25.0%。本试验条件下，施用七水硫酸锌30 kg/hm2，使花生具有较强的抗低温寡照能力及抗病性能，增产显著，可在花生产区推广应用。     

Innovation and gendered negotiations: Insights from six small‐scale fishing communities

There has so far been limited investigation into gender in relation to innovation in fisheries. Therefore, this study investigates how gender relations shape the capacity and motivation of different individuals in fishing communities to innovate. We compare six fishing communities in Cambodia, the Philippines and the Solomon Islands. Our findings suggest that gendered negotiations mediate the capacity to innovate but that wider structural constraints are important constraints for both men and women. Our findings show that men's and women's capacity to innovate is strongly mediated by the behaviour of their marriage partner. Consequently, we argue that gender research from a social relational perspective has an important contribution to make in understanding poor fishing communities where new ways of doing things or new technologies are being promoted.     

Liver histology and histomorphometry in hybrid sorubim (Pseudoplatystoma reticulatum × Pseudoplatystoma corruscans) reared on intensive fish farming

This study aimed to characterize the liver histology and histomorphometry in sorubim hybrid of different categories (nursery, growth and grow‐out) reared on fish farming. The categories were defined considering body weight (BW): nursery category (n = 5): BW = 37.06 ± 6.00 g (31.6–45.3 g); growth category (n = 5): BW = 310.40 ± 53.80 g (242.1–376.4 g) and grow‐out category (n = 5): BW = 874.28 ± 27.59 g (846.2–913.1 g). Liver fragments were processed to paraffin inclusion, and sections were stained by haematoxylin and eosin (H&E), PAS (Periodic Acid Schiff) and Perl's staining to histology, histomorphometry and density volumetric of liver structures; glycogen analysis and to detect ferric irons (Fe³⁺) respectively. The hepatosomatic index decreased between the categories (P < 0.01). The percentage of PAS‐positive hepatocytes in the nursery category was higher (P < 0.05) in relation to the growth and grow‐out categories. The hepatocytes from all fish were positive to Perl's staining. The density volumetric of liver structures did not differ among categories except to blood vessels were higher (P < 0.01) in the nursery and growth. The area (μm²) and perimeter (μm) of hepatocytes, and the area (μm²), perimeter (μm) and volume (μm³) of the nuclei from grow‐out fish were lower (P < 0.01) than those from the nursery and growth categories. Changes in morphometric characteristics of hepatocytes may result from metabolic changes associated with body growth surubins; therefore, these morphometric characteristics of liver tissue can be used as functional biomarkers for the assessment of fish health and nutrition status.     

深水网箱和池塘养殖凡纳滨对虾肌肉营养成分的比较分析

对深水网箱和池塘养殖凡纳滨对虾(Litopenaeus vannamei)肌肉常规营养成分、氨基酸和脂肪酸质量分数进行了比较分析。结果显示:1)网箱养殖对虾肌肉粗蛋白质量分数显著高于池塘养殖(P0.05),而水分、粗脂肪和粗灰分质量分数无显著差异(P0.05);2)肌肉中检测到18种常见氨基酸,网箱养殖对虾肌肉氨基酸总量(TAA)、天门冬氨酸(Asp)、异亮氨酸(Ile)、精氨酸(Arg)和脯氨酸(Pro)质量分数显著高于池塘养殖(P0.05),而必需氨基酸(EAA)、半必需氨基酸(HEAA)、鲜味氨基酸(DAA)和虾味氨基酸(PFAA)两者无显著差异(P0.05);3)肌肉中检测到30种常见脂肪酸,两者的饱和脂肪酸(SFA)、单不饱和脂肪酸(MUFA)、多不饱和脂肪酸(PUFA)、n-3多不饱和脂肪酸(EPA和DHA)质量分数差异显著(P0.05),以网箱养殖对虾较高;两者的棕榈酸(C16∶0)质量分数最高,且网箱养殖对虾EPA+DHA质量分数是池塘养殖的1.90倍。结果表明,网箱养殖对虾营养成分要优于池塘养殖,具有较高的营养价值。     

The effects of diet supplemented with Lactobacillus rhamnosus on tissue parameters of rainbow trout,Oncorhynchus mykiss (Walbaum)

This study was carried out to establish the effects of a 6 week treatment with the diet supplemented with L. rhamnosus in concentrations of 10⁷ CFU g^?1 (G1 group) and 10⁸ CFU g^?1 (G2 group) on the condition expressed by condition factors (Fulton's, Clark's and B), intestinal microbiology, haematological, histological and selected antioxidative parameters of rainbow trout. A significantly higher condition factors were found in G1 group indicating that higher concentration of probiotic (10⁸ CFU g^?1) did not result in the better condition. Cholesterol and urea levels were significantly higher in both G1 and G2 groups, albumin in G1 and creatinine in G2 group with respect to control. A significantly higher liver TBARS level was observed in G2 group. The feeding with supplemented probiont apparently changed the resident microbiota. Three weeks after withdrawal of the supplemented feed, the microflora mostly reverted to the control composition, although L. rhamnosus in faecal matter of fish remained inherent. The epithelial structure of the proximal and distal intestine revealed the increased absorptive area in both treated groups, as well as the increase in the mucin‐secreting goblet cells. The L. rhamnosus‐treated groups demonstrated the capacity for the augmentation of the innate host defence.     

生物絮团对罗氏沼虾体组成和消化酶活性的影响

通过向养殖水体中泼洒糖蜜构建生物絮团养殖模式,分析生物絮团营养组成,并探讨生物絮团对罗氏沼虾体组成和消化酶活性的影响。试验分对照组和试验组(生物絮团组),其中试验组在养殖过程中泼洒糖蜜。试验在室内水泥池内(2 m×2 m×0.6 m)进行,每个处理有3个重复,每个重复225尾虾(0.26 g±0.02 g),试验周期为90 d。养殖过程中不换水,糖蜜的泼洒量根据饲料投喂量进行计算(C/N为20)。结果显示:添加糖蜜能够显著促进生物絮团的形成,到第90天时,试验组的絮团体积达21.22 mL/L;而对照组为6.03 mL/L;试验组絮团粗蛋白含量为29.47%,粗脂肪含量为4.32%,二者均显著高于对照组,而粗灰分含量为11.36%,显著低于对照组;泼洒糖蜜对罗氏沼虾体组成的影响不显著,对照组和试验组肌肉粗蛋白含量分别为21.09%和21.20%,粗脂肪含量分别为2.91%和3.06%;另外,向水体中泼洒糖蜜对罗氏沼虾消化酶活性影响显著。试验组罗氏沼虾肠脂肪酶活性、胃脂肪酶活性和胰脂肪酶活性均显著高于对照组;试验组罗氏沼虾糜蛋白酶活性、胰蛋白酶活性也均显著高于对照组。但泼洒糖蜜对肠淀粉酶、胃蛋白酶、胃淀粉酶、胰淀粉酶和纤维素酶活性没有显著影响。试验表明,生物絮团营养组成丰富,能够有效提高消化酶活性。     

Natural bixin as a potential carotenoid for enhancing pigmentation and colour in goldfish (Carassius auratus)

Discovering natural carotenoids for colour enhancement and health benefits of fish is important to develop new feed formulations. We have purified natural bixin from achiote seeds and evaluated the effect of colour enhancing and pigmentation in goldfish. Varying levels of bixin‐based diets were prepared with 420 g kg^?1 of crude protein and 120 g kg^?1 of lipid content. Our results clearly showed that bixin (0.05, 0.10, 0.20 and 0.60 g kg^?1) based diets significantly (P < 0.05) enhanced the skin and fin colour at 30 and 60 days compared to diet without bixin. Interestingly, diet which contains 0.20 g kg^?1 bixin and commercial feed (with astaxanthin) had similar effect on carotenoid deposition in skin. Moreover, total carotenoid deposition in fin was higher than in skin of all bixin‐containing diets. However, 0.60 g kg^?1 bixin‐containing diet had lower specific growth rate (1.01 ± 0.01) and higher feed conversion ratio (2.05 ± 0.19) compared to the control group. The present results demonstrate that achiote bixin can be successfully used as an alternative natural carotenoid source against synthetic astaxanthin in fish feed. Our data indicate that 0.20 g kg^?1 is a suitable dietary level of bixin to ensure strong pigmentation, acceptable growth and feed utilization in goldfish.     

High fluid shear strain causes injury in silver shark: Preliminary implications for Mekong hydropower turbine design

The Lower Mekong Basin is facing unprecedented threats to fish diversity from hydropower development. There is increasing pressure on developers and construction authorities to design solutions to improve fish survival through turbines, thus protecting the resources in regions being developed for hydropower. A hydraulic characteristic of hydropower turbines with known fisheries impacts is fluid shear stress. Elevated shear stress occurs where rapidly flowing water passes near spillways, screens and within turbine draft tubes. Shear stress can have adverse impacts on fish, but no work has assessed whether this holds true for Mekong River species. A flume was used to determine critical tolerances of silver shark, Balantiocheilos melanopterus (Bleeker), to shear stress rates at a high‐velocity jet which simulated a hydropower turbine. Fish were assessed for injury or mortality following exposure. Results were compared against a no‐shear control. Injury and mortality were greater at higher shear stress exposures. Injuries occurred at all shear exposure levels with mortality at shear levels higher than 600/s. This approach should help design future hydropower turbines if data on other species demonstrate similar results. If the likelihood of adverse impact is high, then shear stress will need to be considered in the design of future hydropower facilities.