Digestion of lipids and carbohydrates from microalgae (Chaetoceros muelleri Lemmermann and Isochrysis aff. galbana clone T‐ISO) in juvenile scallops (Pecten maximus L.)

The digestion of laminaran and neutral lipids from the microalgae Chaetoceros muelleri or mixtures of C. muelleri and Isochrysis galbana clone T‐ISO in 4‐mm scallops Pecten maximus were studied. The microalgae were cultivated at fixed low and high growth rate (5% and 60% of μ_max). The cultivation method clearly affected the lipid and carbohydrate contents, as well as their enzyme activity. Chaetoceros muelleri and T‐ISO grown at low growth rate exhibited a considerably higher content of lipids and carbohydrates than those algae grown at high growth rate. The specific activity of laminaranase and neutral lipase was the highest in C. muelleri grown at the low growth rate. Growth rate of scallops was best when they were fed a mono‐species diet of C. muelleri grown at low growth rate. The growth rate of the scallops was positively correlated to the dietary carbohydrate content as well as to the specific activity of neutral lipase and laminaranase in the diet. The capacity to digest carbohydrates (laminaran) might be limiting for the young scallops, and the content of carbohydrates and enzymatic activities of laminaran in the diet should be considered when formulating an optimal diet, both when the diet consist of live microalgae and formulated diets.     

The importance of manual white blood cell differential counts and platelet estimates in elephant hematology: blood film review is essential

Unique features of elephant hematology are known challenges in analytical methodology like two types of monocytes typical for members of the Order Afrotheria and platelet counts of the comparatively small elephant platelet. To investigate WBC differential and platelet data generated by an impedance-based hematology analyzer without availability of validated species-specific software for recognition of elephant WBCs and platelets, compared to manual blood film review. Blood samples preserved in ethylenediaminetetraacetic acid (EDTA) of 50 elephants (n = 35 Elephas maximus and n = 15 Loxodonta africana) were used. A Mann-Whitney test for independent samples was used to compare parameters between methods and agreement was tested using Bland-Altman bias plots. All hematological variables, including absolute numbers of heterophils, lymphocytes, monocytes, eosinophils, basophils, and platelets, were significantly different (p < 0.0001) between both methods of analysis, and there was no agreement using Bland-Altman bias plots. Manual review consistently produced higher heterophil and monocyte counts as well as platelet estimates, while the automated analyzer produced higher lymphocyte, eosinophil, and basophil counts. The hematology analyzer did not properly differentiate elephant lymphocytes and monocytes, and did not accurately count elephant platelets. These findings emphasize the importance of manual blood film review as part of elephant complete blood counts in both clinical and research settings and as a basis for the development of hematological reference intervals.     

Using genetic tools to inform conservation of fragmented populations of Asian elephants (Elephas maximus) across their range in China

A herd of 15 Chinese elephants attracted international attention during their 2021 northward trek, motivating the government to propose establishment of an Asian elephant national park. However, planning is hampered by a lack of genetic information on the remaining populations in China. We collected DNA from 497 dung samples from all 5 populations encompassing the entire range of elephants in China and used mitochondrial and microsatellite markers to investigate their genetic and demographic structure. We identified 237 unique genotypes (153 females, 84 males), representing 81% of the known population. However, the effective population size was small (28, range 25–32). Historic demographic contraction appeared to account for low haplotype diversity (H_d = 0.235), but moderate nucleotide and nuclear diversity (π = 0.6%, H_e = 0.55) was attributable to post-bottleneck recovery involving recent population expansion plus historical gene exchange with elephants in Myanmar, Lao PDR, and Vietnam. The 5 populations fell into 3 clusters, with Nangunhe elephants differing consistently from the other 4 populations (F_ST = 0.23); elephants from Mengyang, Simao, and Jiangcheng belonged to a single population (henceforth, MSJ), and differed from the Shangyong population (F_ST = 0.11). Interpopulation genetic variation reflected isolation by distance and female-biased dispersal. Chinese elephants should be managed as 2 distinct units: Nangunhe and another combining Shangyong and MSJ; their long-term viability will require restoring gene flow between Shangyong and MSJ, and between elephants in China and neighboring countries. Our results have the potential to inform conservation planning for an iconic megafaunal species.     

Accuracy of pairwise methods in the reconstruction of family relationships, using molecular information from turbot (Scophthalmus maximus)

Many estimators and algorithms have been developed to infer the genealogical relationships from molecular marker data when there is no pedigree information. Most pairwise methods provide estimates in a continuous range that needs to be converted into genealogical relationships (namely full-sibs, half-sibs and unrelated) if there is a previous knowledge of the population structure. Transformations are usually based on arbitrary thresholds, but the possibility of correcting the coancestry estimates via explicit pedigree reconstruction methods has been suggested. Using molecular data for ten highly polymorphic microsatellite loci on a population of turbot (Scophthalmus maximus) with a known genealogy, the behaviour of four pairwise marker-based coancestry estimators and the molecular coancestry has been evaluated. The population consisted on 138 families with 4 full-sib individuals each and one family with 8 full-sib individuals (up to 15 half-sib families were also present due to the sharing of parents between some of the full-sibs families). Our results suggested that transforming the pairwise estimators and the molecular coancestry to family relationships through the establishment of thresholds performs slightly better than the explicit pedigree reconstruction method, when accuracy is measured in a pairwise basis. However, if joint relationships between more than two individuals were tested at a time, the threshold methods led to a high percentage of incongruous triads of full-sib individuals, with Mendelian incompatibilities appearing in congruous full-sib families (more than 70% and 60% in our study, respectively). The explicit pedigree reconstruction approach, due to its nature, is free from such problems. Therefore, the pedigree reconstruction approach seems to be a valuable tool to provide a congruent and compatible pedigree when the pairwise marker-based coancestry matrices or the molecular coancestry need to be transformed.     

大菱鲆生长和耐高温性状的遗传参数估计

对来自40个家系的753尾大菱鲆个体进行耐高温实验,测定其生长和耐高温性状.基于是否考虑全同胞家系效应,建立了两种动物模型,用于估算大菱鲆生长和耐高温性状的遗传参数.利用WOMBAT程序采用平均信息约束极大似然法估计各模型中性状的方差组分,用似然比检验法进行不同模型的差异检验.方差组分分析结果显示,估计体质量遗传参数时,采用模型Ⅰ效果较好,相应遗传力为(0.22±0.09);估计肥满度时,两模型间差异不显著,其遗传力范围为(0.21±0.18)～(0.39±0.11);估计耐热性时,采用模型Ⅰ比较理想,遗传力为(0.026±0.034).表型和遗传相关分析结果表明,体质量与肥满度、耐热性的表型相关系数分别为0.13和0.04,肥满度与耐热性的表型相关系数为0.13;体质量与肥满度、耐热性遗传相关系数分别为0.01和-1.00,肥满度与耐热性的遗传相关系数为0.05.研究结果表明,没有考虑全同胞家系效应的模型Ⅰ是估计大菱鲆生长和耐高温性状遗传参数的较好模型.     

水温和盐度对大菱鲆稚鱼存活的影响

采用突然改变温度和盐度的方法,试验不同水温和盐度对大菱鲆稚鱼存活的影响。试验结果表明,水温为15.0~18.0℃、盐度为28~32,大菱鲆稚鱼存活率最高;用曲线回归的方法求出24 h内存活率与温度的关系式,其回归方程为y=-0.5506x2 18.4970x-53.13,依此得到大菱鲆稚鱼的高、低起始致死温度为26.54℃和7.06℃。依同样方法得到24h内存活率与盐度的回归方程为y=-0.4353x2 23.4226x-213.81,高、低起始致死盐度为37.76和16.05。经t检验显著相关(P<0.01)。     

硒对铜胁迫下大菱鲆幼鱼生长、抗氧化能力及相关基因表达的影响

为探讨硒对铜胁迫下大菱鲆幼鱼生长、抗氧化能力及相关基因表达的影响,以初始体质量为(24.85±0.10) g的大菱鲆为研究对象,通过在饲料中添加五水硫酸铜和亚硒酸钠,配制铜、硒含量分别为(0、0,1 000、0,1 000、2,1 000、4 mg/kg) 4种等氮等能的实验饲料,命名为D1、D2、D3和D4组。养殖实验持续84 d。结果显示:①各组间实验鱼成活率无显著差异;增重率、特定生长率和蛋白质效率均在D2组显著低于D1、D3和D4组;摄食率和饲料系数呈相反趋势,在D2组显著高于其他3组。②幼鱼全鱼和肝脏粗脂肪含量呈先下降后上升趋势;而背肌粗蛋白和粗脂肪含量在各组之间无显著性差异。全鱼、脊椎骨和肝脏中Cu含量在D2组达到最高值;Zn含量则在D4组达到最高值;肝脏中铁含量呈下降趋势,在D3和D4组显著低于D1和D2组。③D2组显著降低了幼鱼血清总抗氧化能力(T-AOC)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)活性;丙二醛(MDA)含量则呈相反趋势,在D2组显著高于D3和D4组;总超氧化物歧化酶(T-SOD)和铜—锌超氧化物歧化酶(CuZn-SOD)活性均在D4组达到最低值。④血清谷丙转氨酶(ALT)、谷草转氨酶(AST)活性均在D2组显著高于其他3组;而血糖浓度、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C)活性均在D2组达到最低值,显著低于D1组。⑤肝脏金属硫蛋白(MT) mRNA相对表达量在D4组达到最高值,显著高于其他3组;谷胱甘肽转移酶(GST)和溶菌酶(LZM) mRNA相对表达量均在D2组显著低于其他3组;热休克蛋白70 (HSP70) mRNA相对表达量则在D2组显著高于其他3组。综上所述,在本实验条件下,饲料中添加硒(2～4 mg/kg)可缓解高铜(1 000 mg/kg)胁迫导致的鱼体生长缓慢等症状,并可调控鱼体的抗氧化能力、生理代谢及相关基因表达量,进而对鱼体在高铜胁迫下的内环境稳态恢复具有一定的促进作用。     

不同贮藏温度下大菱鲆品质变化及货架期预测模型的建立

为探究大菱鲆(Scophthalmus maximus)在不同贮藏温度下品质特性与货架期的关系,将大菱鲆贮藏在-3℃、0℃、4℃、10℃和15℃温度下,测定其感官品质、挥发性盐基总氮(TVB-N)、菌落总数、硫代巴比妥酸值(TBA)、电导率的变化,并且观测肌肉的微观结构;采用低场核磁共振技术(Low-field nuclear magnetic resonance, LF-NMR)分析鱼肉中水分迁移状况,并且建立了TVB-N及菌落总数与贮藏时间和温度的动力学模型。研究发现,随着贮藏时间的延长,5种不同贮藏温度下鱼肉中不易流动水均减少,货架期终点各贮藏温度下的样品相对于新鲜鱼肉,其肌纤维结构均由紧密变得疏松;TVB-N和菌落总数变化预测模型中的活化能和指前因子分别为79.50、75.07 kJ/mol和1.3×10^14、7.62×10^12。选用10℃进行验证性试验,结果显示实测值与预测值相对误差在10%以内。因此,可根据TVB-N值及菌落总数对大菱鲆贮藏在-3℃~15℃的货架期进行实时预测。     

还原型谷胱甘肽对大菱鲆生长及抗氧化能力的影响

在基础饲料中添加不同剂量的还原型谷胱甘肽,添加量分别为0、100、200、400、600 mg/kg,投喂初始体质量为(23.08±0.09) g的大菱鲆,8周后测定还原型谷胱甘肽对大菱鲆生长及抗氧化能力的影响。试验结果表明,饲料中还原型谷胱甘肽添加量为200 mg/kg时,大菱鲆的质量增加率和特定生长率显著高于其他组(P<0.05)。饲料中添加还原型谷胱甘肽对大菱鲆肝脏中丙二醛含量、总抗氧化能力、超氧化物歧化酶活力、谷胱甘肽过氧化物酶活力均无显著影响(P>0.05)。随着还原型谷胱甘肽添加量的增加,大菱鲆肝脏中丙二醛含量呈先降后升的趋势,对照组最高,200 mg/kg试验组最低;大菱鲆肝脏中总抗氧化能力、超氧化物歧化酶活力和谷胱甘肽过氧化物酶活力均呈先升后降的趋势,200 mg/kg试验组最高。200 mg/kg试验组和400 mg/kg试验组大菱鲆肝脏中还原型谷胱甘肽含量显著高于对照组(P<0.05)。200 mg/kg试验组大菱鲆肝脏中谷胱甘肽硫转移酶活力和谷胱甘肽还原酶活力显著低于对照组(P<0.05)。根据回归分析,确定大菱鲆饲料中还原型谷胱甘肽的最适添加量为189.70 mg/kg。     

大菱鲆AFLP分析体系的建立

本研究构建了大菱鲆扩增片段长度多态性(AFLP)分析体系,对DNA提取、双酶切反应、连接反应、预扩增反应、选择性扩增反应和银染等步骤进行了分析。结果表明,用EcoRI/MseI双酶切、核心序列加3个选择性碱基的选择性扩增引物、1∶6(w∶w)的EcoRI端与MseI端选择性扩增引物比和20μl选择性扩增体积,可得到十分稳定的结果。所得产物经电泳和银染后可获得条带清晰、背景干扰小的图像。该体系的构建为AFLP技术在大菱鲆相关研究中的应用奠定了基础。