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.     

盐度和温度对大菱鲆幼鱼抗氧化酶活性的影响

采用两因素交叉分组的方法,研究了温度和盐度对大菱鲆Scophthalmus maximus幼鱼行为活动、摄食、存活情况以及体表黏液、血液、鳃、肝脏中抗氧化酶活力的影响.结果表明:水温(17、20、23、25、28℃)和盐度(5、10、20、30、40)对大菱鲆幼鱼摄食、死亡率有显著影响(P＜0.05),在水温为17、20℃与盐度为20、30的组合条件下,大菱鲆幼鱼的摄食积极性以及存活率较高,显著高于其他组(P＜0.05);温度和盐度对大菱鲆幼鱼SOD、CAT与GP-X活力有显著影响(P＜0.05);当温度一定,盐度变化的条件下,肝脏、鳃、血清和黏液中的各种酶活力在正常海水盐度(30)时活力较低,并随盐度由30起降低或升高而均出现升高的趋势;当盐度一定,温度变化的条件下,随着水温的升高,各组织中的酶活力变化规律不显著;通过分析温度与盐度交互作用对各组织不同酶活的变化以及生命表征现象,得出了最适生存阈值,即大菱鲆幼鱼的适宜生活水温为17～20℃,适宜生活盐度为20～30.     

养殖大菱鲆肠炎病的流行病学和病原学研究

[目的]肠炎病是目前大菱鲆养殖行业中比较常见的疾病之一,该病感染率高,传播迅速。[方法]从山东半岛3个不同的养殖厂肠炎病发病大菱鲆体内分离致病菌,对其进行了常规生理生化特征测试和16S rRNA基因序列对比研究。[结果]从发病大麦鲆体内共分离到了株优势细菌。理化特征分析结果显示这3株菌中有2株分别与溶藻弧菌(Vibrio alginolyticus)和大菱鲆弧菌(Vibrio scophthalmi)表型特征非常相似,另1株细菌被证实为非弧菌属的细菌,具体定种尚需进一步研究。分析这2株弧菌属细菌的16S rRNA基因序列并构建了系统发育树,结果表明这些菌株与溶藻弧菌(Vibrio alginolyticus)和大菱鲆弧菌(Vibrio scophthalmi)的亲缘关系最近。[结论]从发病大菱鲆体内分离菌株的2株细菌被鉴定为溶藻弧菌(Vibrio alginolyticus)和大菱鲆弧菌(Vibrio scophthalmi)。     

神经型一氧化氮合酶在大菱鲆脑组织中的分布及定位

[目的]探明神经型一氧化氮合酶(nNOS)在大菱鲆(Scophthalmus maximus)脑组织中的分布情况,为揭示大菱鲆脑组织中一氧化氮(N0)的生理功能提供形态学资料.[方法]分别采用NADPH-d组织化学染色法和免疫组织化学法对大菱鲆脑组织中的nNOS进行定位研究.[结果]NADPH-d组织化学染色结果显示,大菱鲆大脑皮质中有蓝色的神经元和神经纤维存在.神经元呈梭形、锥形等形状,神经纤维呈串珠状且无序交织;小脑中NOS阳性神经元在分子层分布稀疏,在颗粒层分布密集,浦肯野细胞胞核淡染.经免疫组织化学法染色后,可观察到大脑皮质中nNOS免疫组化反应阳性物质呈深棕色;小脑皮质的分子层、颗粒层及浦肯野细胞层均有nNOS阳性神经元分布,浦肯野细胞呈免疫组织化学阳性反应.[结论]大菱鲆脑组织中的NOS类型主要限于nNOS,且nNOS在神经活动中发挥重要作用,也进一步证实N0在不同物种中具有一些共同的作用,但不同物种或相同物种不同组织中NO分布及含量存在差异.     

鱼用三联口服疫苗的制备方法及其对大菱鲆的免疫效果

为探究海水养殖中安全可靠、经济适用的疫苗接种方法,以药用淀粉、麦芽糊精,以及经0.5%甲醛灭活的溶藻弧菌Vibrio alginolyticus、副溶血弧菌V.parahemolyticus和迟缓爱德华氏菌Edwardsiella tarda为主要原料,采用挤出-滚圆法制备鱼用三联口服疫苗,并用其对大菱鲆Scophthalmus maximus免疫,分别在免疫0、7、14、21、28 d时测定溶藻弧菌、副溶血性弧菌和迟缓爱德华氏菌特异性抗体,以1×109cfu/m L活菌(3种菌液比例为1∶1∶1)进行攻毒试验并计算免疫保护率。结果表明,药用淀粉150 g、麦芽糊精50g、菌液60 m L、挤出速度200 r/min、滚圆时间60 s时,为比较成熟的三联口服疫苗制备工艺,用该疫苗免疫大菱鲆后,3种细菌的大菱鲆血清和后肠黏膜抗体效价均显著高于对照组,且免疫保护率达50%以上。     

周期性饥饿再投喂对大菱鲆幼鱼生长、生化组成和能量收支的影响

在水温为17～19℃条件下,分别以鲜杂鱼(x)和配合饲料(p)为饵料,采用一直投喂(对照组)和周期性饥饿1 d再投喂6 d(S1F6)、饥饿2 d再投喂5 d(S2F5)3种投喂方式,研究投喂不同饵料时周期性饥饿再投喂对大菱鲆Scophthalmus maximus幼鱼(体质量为7.80 g±0.01 g)生长、生化组成和能量收支的影响,试验共进行42 d。结果表明:投喂配合饲料时,S1F6p和S2F5p组均具有完全补偿生长现象;投喂鲜杂鱼时,S1F6x组具完全补偿生长现象,S2F5x具部分补偿生长现象;投喂配合饲料和鲜杂鱼时,周期性饥饿再投喂处理的大菱鲆幼鱼均通过饲料转化效率和摄食率共同提高来实现其补偿生长,且各试验组的鱼体成分和对照组均无显著性差异(P>0.05);与对照组相比,试验处理组幼鱼的生长能比例均有所增加,而代谢能比例均有所减少;无论是处理组还是对照组,鲜杂鱼的投喂效果均优于配合饲料。     

饲料Vc对大菱鲆非特异性免疫力的影响

X用维生素C（Vc）含量为0～2500mg／kg的配合饲料连续投喂大菱鲆70d后，测定大菱鲆血液白细胞的总数、吞噬活性、血清溶菌酶活性和总补体活性。结果表明，饲料中Vc含量增加到250mg／kg，白细胞总数明显增加，进一步提高Vc含量，白细胞总数没有显著变化；血清溶菌酶活性在Vc含量为250mg／kg时最高，其他各组无显著性差异；添加Vc的各试验组的总补体活性明显地高于对照组，但是添加Vc的各组之间的总补体活性没有显著性差别；Vc含量对白细胞的吞噬活性没有影响。     

养殖大菱鲆肠道中大菱鲆弧菌的分离鉴定及药敏试验

2014年11月,大连地区某大菱鲆养殖场发生病害并伴有死亡,主要症状为吻部下颌明显出血,腹腔积水,肝脏充血,肠道出血有白便。从患病大菱鲆肠道中分离出1株可在硫代硫酸盐—柠檬酸盐—胆盐—蔗糖琼脂培养基上生长的菌株HZ-C1,人工回接感染试验证明其对健康大菱鲆具有较强致病性。通过形态学观察、16SrDNA序列同源性分析及生理生化试验确定菌株HZ-C1为大菱鲆弧菌。药敏检测结果显示,大菱鲆弧菌HZ-C1对氯霉素、丁胺卡那及头孢曲松等11种抗生素敏感,但对青霉素、四环素及诺氟沙星等15种抗生素耐药。本研究为大连地区养殖大菱鲆细菌性病害防治提供了一定病原学依据。