Impacts of dietary supplementation of bamboo vinegar and charcoal powder on growth performance,intestinal morphology,and gut microflora of large-scale loach Paramisgurnus dabryanus

Although accumulating data demonstrate that dietary supplementation of bamboo vinegar and charcoal powder(BVC)can significantly strengthen the immunity and boost the growth of domestic animals,its application potency still awaits verification in commercial fish species such as the large-scale loach Paramisgurnus dabryanus.The impacts of 90-day dietary supplementation of 1%and 2%BVC on survival and growth performance of the loach,and on the intestinal morphological characteristics and gut microflora were analyzed.Our data show that the large-scale loach supplied with BVC at the experimental doses had significant higher survival rates and better growth performance(indicated by greater weight gain(1.13–1.14 times),higher specific growth rate(1.04 times),and lower feed conversion ratio(0.88–0.89 times))compared to that of the control(P<0.05).Histological examination revealed significant longer villus(3.22–5.54 times),deeper crypt(1.77–1.87 times),and thicker muscle(1.59–3.17 times)in the intestines of large-scale loach fed with BVC(P<0.05).Furthermore,we found that the gut microflora consisted of significantly fewer proportions of potential pathogenic bacterial species(Aeromonas veronii and Escherichia coli),but significantly greater proportions of beneficial microbes(Lactococus raffinolactis and Faecalibacterium prausnitzii).Therefore,dietary intake of BVC can promote intestinal tract development and optimize gut microflora,by which the survival and growth of large-scale loach may be improved.

Statistic Morphology Difference between Male and Female Individuals of Paramisgurnus dabryanus Sauvage

[Objective] The aim was to compare the morphology difference between male and female individuals of Paramisgurnus dabryanus Sauvage. [Method] The comparison of 10 morphological characteristics between male and female individuals of P. dabryanus was conducted with principal component analysis and t-test methods. [Result] The results of t-test indicated that three of 10 morphological characteristics showed extremely significant difference（P0.01）and one of 10 morphological characteristics was significantly different（P0.05）. Principal component analysis revealed four principal components,the contribution rate of which was 31.151%,18.844%,12.324% and 10.169%,respectively. The cumulative contribution rate had reached 72.487%. The morphological variations between male and female individuals were not significant,which was consistent with that of t-test. [Conclusion] The result in this study had provided theoretical basis for the breeding of P. dabryanus.

Comparative Study on Autumn Propagation Performances of Paramisgurnus dabryanus and Misgurnus anguillicaudatus in Poyang Lake

Preliminary comparative study was conducted on autumn propagation per- formances of Paramisgumus dabryanus and Misgurnus anguillicaudatus. The count- able and measurable properties, fatness, mature coefficient, relative fecundity, as well as the fertilized and the hatching rates were measured. The results showed that there was no significant difference between the fatness of the two species （P〉0.05） and between the fatness of male and female Paramisgumus dabryanus （P〉0.05）, while significant difference existed between male and female Misgurnus anguillicauda- tus （0.01〈P〈0.05）; the mature coefficients of Paramisgurnus dabryanus and Misgurnus anguillicaudatus were of 6.56% and 11.36%, respectively; the relative fecundities of Paramisgurnus dabryanus and Misgurnus anguillicaudatus were of 115.70 eggs/g and 147.69 eggs/g, respectively; the fertilized rates of Paramisgurnus dabryanus and Mis- gurnus anguillicaudatus were of 90.62% and 97.52%, respectively; and the hatching rates of Paramisgumus dabryanus and Misgurnus anguillicaudatus were of 92.45% and 97.16%, respectively. It was concluded that Misgumus anguillicaudatus was bet- ter than Paramisgurnus dabryanus in breeding performance in autumn.

Morphological changes and allometric growth in hatcheryreared Chinese loach Paramisgurnus dabryanus(Dabry de Thiersant, 1872)

The length-weight relationship and allometric growth patterns of hatchery-reared Chinese loach, Paramisgurnus dabryanus(Dabry de Thiersant, 1872), were determined from hatching to 60 days after hatching. A best power model was estimated for the length-weight relationship during the early life stages. Positive allometric growth for the head segment, trunk length, tail length and eye diameter was also found in the early life stages, while body depth, tail depth, tail fi n length, pectoral fi n length and barbel length displayed a negative coeffi cient. During the subsequent early developmental stage, the growth coeffi cients showed a clear and common tendency towards isometry for all measured body ratios. The allometric growth changes in Chinese loach during the early stage are possibly the result of selective organogenesis directed towards survival priorities.

PCR Amplification of Sox Genes in Misgurnus anguillicaudatus and Paramisgurnus dabryanus(Cypriniform;Cobitidae)

Using the primers which specially amplify the conservative motif of Human SRYgene,we stud ied the PCR amplification of Sox genes in genomic DNA of two species of mud loach Misgurus anguillicau-datus and Paramisgurnus dabryanus.Four bands with the length of 200,550,940 and 1000 bp respectively,were presented in the PCR products of Misgumus anguillicaudatus.Three bands with the length of 200,550 and 900 bp were presented in that of Paramisgumus dabryamus.Southern blotting results indicated that the 200 and 550 bp bands are specially positive.There is no difference between male and female individuals as well as between these two species.

Transcriptomic analysis of regenerated skins reveals key genes involved in skin immune response and air-breathing of loach(Paramisgurnus dabryanus)

Fish skin acts as the first immune barrier against pathogens from external environments.Recently,fish skin immunity research has gradually become a hot topic in aquaculture disease control.Interestingly,the skin can do air-breathing in some bimodal respiration fish.However,there is no comprehensive understanding of these two functions.In this study,the skin regeneration of loach(Paramisgurnus dabryanus)was investigated through morphological and histological observations.Then,original skins(OS)and the regenerated skins(RS)when their capillaries were the most abundant during healing,were collected for transcriptomic analysis.285,899,386 clean reads with a total length of 42.34 Gb were obtained.A total of 1282 differentially expressed genes(DEGs)were detected,including 1030 DEGs up-regulated and 252 DEGs down-regulated in the comparison of RS vs.OS.Fc gamma R-mediated phagocytosis(containing gelsolin(Gsn)),chemokine signaling pathway(containing C-C motif chemokine ligand 28(Ccl28)),and B cell receptor signaling pathway(containing CD80 molecule(Cd81))were closely related to skin immune response of the loach.PI3K-Akt signaling pathway[containing fibronectin 1(Fn1)and lysyl oxidase-like 2b(Loxl2b)],TGF-beta signaling pathway[containing thrombospondin 1(Thbs1)],ECM-receptor interaction[containing integrin alpha 7(Itga7),Itgb8,Itgb10 and Itgb5],and dilated cardiomyopathy[containing laminin subunit alpha1(Lama1)]were closely associated with skin air-breathing of the loach.This study is conducive to explore the molecular mechanisms of skin immune response during its regeneration and air-breathing of bimodal respiration fish.This study will benefit for the aquaculture of P.dabryanus and its related species.

Effects of different protein and lipid levels on the growth performance and intestinal microflora of loach(Paramisgurnus dabryanus)

The aim of this study was to examine the effects of dietary protein and lipid levels on the growth per-formance and homeostasis of the intestinal flora in Paramisgurnus dabryanus.An 8-week 3×3 two-factorial experiment was conducted to investigate the interaction between dietary crude protein(CP:30%,35%,40%)and ether extract(EE:6%,10%,14%)on the growth rate and the intestinal microflora of P.dabryanus.A total of 2,160 fish(5.19±0.01 g)were randomly allotted to 36 aquariums each with 60 fish.Fish were fed the experimental diet twice daily.Results revealed that weight gain rate(WGR),specific growth rate(SGR),protein efficiency ratio and net protein utilization significantly increased when increasing protein levels from 30%to 40%(P<0.05).Both WGR and SGR enhanced first but reduced thereafter with maximum value at 10%lipid level as dietary lipid increased from 6%to 14%(P<0.05).Significant interactions between protein and lipid were found with feed conversion rate,lipid efficiency ratio and net lipid utilization(P<0.05).At the phylum level,Proteobacteria and Actinobacteria were the dominant bacteria;at the genus level,Burkholderia-Caballeronia-Paraburkholderia was the dominant bacteria.Fish fed the diet containing 10%lipid had a higher abundance of Proteobacteria and unclassi-fied_f_Eenterobacteriaceae than those fed the 14%lipid diet,and a higher abundance of Rhodobacter than those fed the 6%lipid diet(P<0.05).Analysis of the predicted functions showed that metabolism in the intestine of fish in the CP40EE10 group was more active than that in CP30EE14 group.Polynomial regression analysis found that a diet containing 40.87%protein and 9.88%lipid can be considered optimal for P.dabryanus.

三种常用稻田农药对大鳞副泥鳅的急性毒性研究

为研究吡蚜酮、稻杰和二甲四氯钠三种常用稻田农药对大鳞副泥鳅(Paramisgurnus dabryanus)急性毒性的影响,选择体长约2.74 cm、体重约5.57 g的大鳞副泥鳅作为受试对象,分别采用40 mg/L、80 mg/L、180 mg/L、280 mg/L和380 mg/L药物浓度进行急性毒性试验。结果显示,吡蚜酮的安全浓度最高,二甲四氯钠次之,稻杰最低,而三者的96 h半致死浓度分别为280 mg/L、8 mg/L和90 mg/L,安全浓度分别为28 mg/L、0.8 mg/L和9 mg/L。研究结果表明,大鳞副泥鳅对这3种农药都表现出较高的敏感性,其敏感性依次为稻杰>二甲四氯钠>吡蚜酮。该研究为稻田养殖大鳞副泥鳅过程中的农药使用浓度提供了理论基础。

Unique Structure and Mechanical Property of Dabryanus Scale

The structure and mechanical property of Dabryanus scale were investigated. Environmental Scanning Electron Micros- copy （ESEM） and Transmission Electron Microscopy （TEM） were used to observe the scale structure. The scale has a discrete osseous layer which is composed of boat-like unit （due to the shape）. The osseous layer is embedded into fibrillary layer, and the largest thickness ratio of osseous layer to fibrillary layer is about 7, much higher than those of other reported fish scales. Nanoindentation test was used to investigate mechanical property of the scale. Elastic modulus （E,） and hardness （H） of Dabryanus scale are much lower than those of other reported fish scales. Scale models were proposed and the principles of materials mechanics were used to analyze the mechanical properties of the Dabryanus scale. The boat-like unit in osseous layer makes the scale flexible while keeping a high thickness ratio of osseous layer to fibrillary layer, which increases the flexibility and protection capacity of the scale This study may provide new strategy for the design of flexible armor or flexible devices.

Characterization of hematopoiesis in Dabry's sturgeon(Acipenser dabryanus)

The hematological indices of Dabry's sturgeon(Acipenser dabryanus)were investigated in this paper.The results showed that the count of Red blood cell(RBC)was(0.60±0.02)
1012/L,and the blood glucose(GLU)was 9.32±1.00 mmol/L.Glutamic-pyruvic transaminase(ALT)and glutamic-oxalacetic transaminase(AST)were 13.75±6.65 U/L and 359.50±40.07 U/L,respectively.Smears of peripheral blood and imprints of four tissues were studied in the trial with Wright's staining method.The results indicated that Dabry's sturgeon's blood cells could be divided into five cell types,and included erythroid,granuloid,monocytoid,lymphoid and other cells.In addition,the developmental stage of each cell lineage could be subdivided into:primitive,young and mature.In the three developmental stages,the size of each cell type was similar and gradually decreased from primitive,young to mature blood cells.Among the blood cells,the monoblast and macrophage were the biggest,while the thrombocyte was the smallest.All stages of the primitive cells and young cells were observed in head kidney and all cell types,except granuloblasts,were identified in the head kidney.In contrast,primitive cells and young cells were not observed in the spleen or liver,and few orthochromatic erythroblasts were found in peripheral blood.It can be concluded that head kidney and trunk kidney are the primary organs of hematopoiesis in the Dabry's sturgeon.

干旱胁迫对大鳞副泥鳅免疫指标的影响

为探究气呼吸型鱼类在养殖运输中常遭遇的短时干旱胁迫对其免疫应答的影响,本实验以大鳞副泥鳅(Paramisgurnus dabryanus)为研究对象,将其暴露于空气中12、24、48、72、96 h后检测血清和肝脏免疫指标的变化。结果显示:空气暴露显著增加了大鳞副泥鳅血清总蛋白、白蛋白、球蛋白和免疫球蛋白M含量,但对血清和肝脏溶菌酶活性无显著影响。补体C3和补体C4含量均呈先升后降的变化趋势。血清热休克蛋白70和热休克蛋白90含量在暴露48 h分别升高至基础水平的3.3倍和2.0倍。血清炎症细胞因子水平随着暴露时间的延长呈现先升后降的趋势;空气暴露12 h,肝脏肿瘤坏死因子-α和白细胞介素-1β上升至基础水平的1.4倍,白细胞介素-6和白细胞介素-12上升至1.5倍。上升的免疫指标表明短期(0~48 h)的干旱胁迫对大鳞副泥鳅有明显的免疫刺激作用,随后多数免疫指标在实验后期(48~96 h)下降,表明持续的干旱胁迫导致免疫刺激转为免疫抑制作用。

大鳞副泥鳅皮肤抗氧化酶活性的昼夜节律研究

昼夜节律是生物生理学研究重点,用以揭示物种特有的活动规律。该试验首次以大鳞副泥鳅(Paramisgurnus dabryanus)为研究对象,探究其皮肤抗氧化酶活性的昼夜节律变化。将300尾泥鳅随机分成三组,暂养阶段结束后,取皮肤组织检测各种抗氧化酶活性。试验结果表明,大鳞副泥鳅皮肤组织的总抗氧化能力(T-AOC)、超氧化物歧化酶(T-SOD)、谷胱甘肽转移酶(GST)和谷胱甘肽过氧化物酶(GSH-Px)活性的峰值都出现在夜晚,过氧化氢酶的活性峰值出现在08:00。同时,丙二醛的峰值出现在12:00,谷值出现在04:00,差异性显著(P<0.05)。研究发现,大鳞副泥鳅皮肤组织的抗氧化酶活性呈现明显的昼夜节律,并且在夜晚具有较高的抗氧化能力。

饲料蛋白水平对达氏鲟幼鱼生长性能、体组成、消化酶活性以及血液生化指标的影响

为了解达氏鲟（Acipenser dabryanus）幼鱼适宜的饲料蛋白质水平,试验选取平均初始体重为（400.68±8.55）g的健康达氏鲟幼鱼为试验对象,在室内流水系统中进行为期8周的摄食生长实验。试验鱼随机分为5组,每组3个重复,每个重复30尾鱼,分别饲喂蛋白质水平为30%、35%、40%、45%、50%的5种等脂等能饲料。试验期间,水流量为6 L/min,水温（24.0±1.0）℃,溶氧大于6.0 mg/L,氨氮小于0.02 mg/L。结果显示：增重率随饲料蛋白水平的增加呈现先升高后降低的趋势,饲料系数呈现相反的趋势。蛋白质效率随饲料蛋白水平的增加呈先平缓后下降的趋势。各试验组间肥满度无显著差异。肝体比和脏体比随饲料蛋白水平的增加而下降。各试验组间甘油三酯和总胆固醇含量无显著差异,饲料蛋白水平为50%的试验组的谷丙转氨酶（ALT）和谷草转氨酶（AST）活力显著高于其他试验组。本试验中胃蛋白酶活力与饲料蛋白水平呈正相关关系,淀粉酶活力呈负相关关系,脂肪酶在各试验组无显著差异。鱼体蛋白和脂肪含量均随饲料蛋白水平的增加而增加。分别对增重率、饲料系数这二者与饲料蛋白水平之间的关系进行二次回归分析,得出达氏鲟幼鱼饲料中适宜的蛋白水平为41.5%-42.97%。

几种常用水产药物对大鳞副泥鳅苗种的急性毒性试验

以规格(1.53±0.32)g重的大鳞副泥鳅(Paramisgurnus dabryanus)苗种为试验对象,采用静态急性毒性试验法,进行了硫酸铜、聚维酮碘、敌百虫、甲醛的急性毒性试验。结果表明,硫酸铜24、48、96 h的半致死量分别为1.23、0.55、0.40 mg/L,安全浓度为0.03 mg/L;聚维酮碘24、48、96 h的半致死量分别为429.58、396.36、388.47 mg/L,安全浓度为101.23 mg/L;敌百虫24、48、96 h的半致死量分别为16.72、7.94、4.80 mg/L,安全浓度为0.54 mg/L;甲醛24、48、96 h的半致死量分别为110.00、96.59、45.45 mg/L,安全浓度为22.35 mg/L。结论:硫酸铜的毒性最大且安全浓度远低于常用浓度,应禁用;敌百虫的安全浓度处于常用浓度范围内,但使用时需控制使用剂量;聚维酮碘和甲醛可按常规浓度安全使用。

敌百虫、溴氰菊酯对大鳞副泥鳅仔鱼及多刺裸腹溞的急性毒性实验

采用静水实验法在p H值为8.2±0.1、水温(25±2)℃,溶解氧大于5 mg/L条件下,以5日龄大鳞副泥鳅(Paramisgurnus dabryanus)仔鱼和多刺裸腹溞(Moina macrocopa)为材料,研究了敌百虫和溴氰菊酯2种常用药物对大鳞副泥鳅仔鱼和多刺裸腹溞的急性毒性。结果显示:敌百虫对5日龄大鳞副泥鳅仔鱼和多刺裸腹溞的安全浓度分别为0.400 0 mg/L、0.048 9 mg/L;溴氰菊酯对大鳞副泥鳅仔鱼和多刺裸腹溞的安全浓度分别为0.160 0μg/L、0.004 2μg/L。结果表明,敌百虫急性毒性要显著性强于溴氰菊酯。

真泥鳅和大鳞副泥鳅怀卵量的比较分析

研究了真泥鳅(Misgurnus anguillicaudatus)和大鳞副泥鳅(Paramisgurnus dabryanus)体长与怀卵量的关系。结果表明,两种泥鳅的平均绝对怀卵量都随体长的增加有明显的增加趋势,体长在10 cm以下时,两种泥鳅的平均绝对怀卵量没有显著差异;体长在10～25 cm时两种泥鳅的平均绝对怀卵量均存在显著差异;同一体长段真泥鳅的怀卵量高于大鳞副泥鳅。

达氏鲟gdf11基因cDNA的克隆及组织表达特征分析

本研究克隆了达氏鲟(Acipenser dabryanus)生长转化因子gdf11(growth differentiation factor 11)基因cDNA。达氏鲟gdf11基因cDNA序列全长1 298 bp(不包括Poly A),开放阅读框为1 191 bp,编码396个氨基酸,5非编码区长19 bp;3非编码区长88 bp。通过Signal P软件预测含有N端21aa的信号肽。组织表达特征分析结果显示,gdf11在7种组织中均有表达,在眼和鳃中的表达量较高;不同水流条件下,鳃和心脏gdf11的表达量有显著性差异,静水中鳃gdf11表达量是流水(流速27.0±3.0 cm/s)的4.70倍,而心脏中的表达量是流水的2.72倍。这暗示了gdf11可能在呼吸代谢中发挥功能。

南四湖大鳞副泥鳅人工繁殖与批量生产技术研究

根据对大鳞副泥鳅(Paramisgurnus dabryanus)繁殖生产实践的探索,总结了一种从产前准备到催熟、催产、孵化和出苗五个生产阶段的苗种批量生产的方法。通过受精卵生物学实验、不同雌雄比生产性试验,提出雌雄分离单性催熟、混合催产素集中注射、同步催产、卵与亲鳅快速分离和发塘标志性形态特征判定等多项关键的技术措施,实现了苗种快速批量生产。三组不同雌雄比的生产试验表明:繁殖亲鳅300组,雌雄比为1∶0.85时,孵化率为70.31%,出苗量最高,出苗152.7万尾。该研究为加快大鳞副泥鳅产业发展,规范人工繁殖和苗种批量生产提供依据。

17β-雌二醇对大鳞副泥鳅生长和性腺分化的影响

采用短期连续投喂的方式,研究了不同浓度17β-雌二醇(0.25、0.50、0.75和1.00 mg/kg)对大鳞副泥鳅(Paramisgurnus dabryanus)生长和性腺分化的影响。结果显示:药物处理对大鳞副泥鳅成活率无显著影响;短期内(60 d)17β-雌二醇能显著促进大鳞副泥鳅的生长,其中低浓度(0.25 mg/kg)作用效果最明显;投喂60 d后,各浓度组的雌性比例(66.11%、71.67%、72.78%和71.11%)均明显高于对照组(56.67%),其中以0.75mg/kg组雌性比例最高。结果表明17β-雌二醇能促进大鳞副泥鳅未分化性腺向卵巢发育。

三苯基锡和聚苯乙烯微塑料联合暴露对大鳞副泥鳅的急性毒性效应

以大鳞副泥鳅(Paramisgurnus dabryanus)为受试生物,三苯基锡和聚苯乙烯微塑料(R=200 nm)为染毒物质,进行三苯基锡单独暴露以及与聚苯乙烯微塑料联合暴露的毒性试验,揭示三苯基锡对大鳞副泥鳅的急性毒性效应规律以及聚苯乙烯微塑料对三苯基锡急性毒性的调节作用。在水温(20.0±1.0)℃、溶氧充足(≥6.5 mg/L)条件下,采用半静态水生生物急性毒性实验方法,根据预实验结果设置6个浓度组(0.01、0.016、0.024、0.037、0.058和0.09 mg/L)和一个空白对照组,聚苯乙烯微塑料浓度为1 mg/L。结果显示,三苯基锡单独暴露对大鳞副泥鳅的24、48、72和96 h的半致死浓度(LC50)分别为0.086、0.037、0.026和0.019 mg/L,安全浓度(SC)为0.002 mg/L,药物蓄积程度系数(MAC)随实验时间延长呈较快下降趋势。三苯基锡与聚苯乙烯微塑料联合暴露实验结果表明,中高浓度组(0.037、0.058 mg/L)100%的死亡率较三苯基锡单一毒性结果均有延迟,而高浓度组(0.09 mg/L)无显著变化,表明聚苯乙烯微塑料对三苯基锡的急性毒性具有延缓作用且存在浓度阈值。