鲈鲤(Percocypris pingi)染色体核型及C-带分析

鲈鲤(Percocypris pingi)为长江上游特有鱼类及四川省重点保护鱼类,为验证其倍性及丰富其细胞遗传学内容,采用植物性血凝素(PHA)体内注射肾细胞直接制片法研究鲈鲤的核型,采用改进的BSG法研究其C-带.结果显示,鲈鲤的染色体数目为98条,其核型公式为4n=98=42m+30sm+10st+16t,臂数NF=170.本研究发现,鲈鲤有16条端部着丝粒染色体,未发现小染色体及其他异型性染色体.鲈鲤C-带带纹较丰富,显示着丝粒带、端粒带和染色体居间带3种类型的C-带,部分染色体同时含有多种类型的C-带着色,如m2、st4同时含有以上3种C-带带纹类型.鲈鲤染色体异染色质含量高达(58.824±0.034)％.本研究再次确认鲈鲤为四倍体,同时指出其染色体具有一定的“可塑性”,反应出鲈鲤对于地区特化的生境具有较强的适应性.     

鲈鲤苗种培育及苗种生长特性的研究

为研究鲈鲤(Percocypris pingi)苗种的生长特性,开发鲈鲤育苗技术,采用水泥池流水单养模式,在自然水温条件下,开展了10日龄至110日龄鲈鲤苗种培育试验。经过100 d的培育,鲈鲤鱼苗体质量从(46.45±4.03)mg增加到(2600.09±105.18)mg,平均体长从(16.22±0.46)mm增长到(50.78±2.97)mm,平均成活率为82.00%;体质量变异系数在4.05%~24.80%,体长变异系数在2.49%~7.32%,体质量变异系数大于体长变异系数;体质量变异系数整体呈现先升高后降低的变化趋势,体长变异系数整体变化较为平缓,在整个培育阶段,鲈鲤种群生长离散呈现从加剧到减弱的变化规律;体质量特定生长率在(0.91±0.49)%/d~(7.75±2.18)%/d,体长特定生长率在(0.47±0.29)%/d~(1.94±1.46)%/d,体质量特定生长率大于体长特定生长率;在水温16.8~23.0℃条件下,鲈鲤生长状况良好,特定生长率相对较高,当水温高于23.0℃之后,特定生长率呈降低趋势,表明16.8~23.0℃是鲈鲤鱼苗适宜的生长水温范围;肥满度变化范围在(1.09±0.10)~(2.85±0.18)g/cm^3,肥满度变化呈现先升高后降低的变化趋势;体长(x)和体质量(y)的拟合方程为y=0.0015x^3.7566(R^2=0.9806),b值为3.7566;全长(y)和体长(x)的拟合方程为y=1.223x-0.5055(R^2=0.9908)。试验结果表明,鲈鲤苗种在水泥池流水单养模式及自然水温条件下生长状况良好,16.8~23.0℃是鲈鲤苗种的适宜生长水温。     

基于PAE编码系统的短须裂腹鱼行为谱

短须裂腹鱼(Schizothorax wangchiachii)是长江上游特有鱼种。近年来,由于受过度捕捞、水电开发等人类活动影响,短须裂腹鱼自然资源量不断下降。2015年11月—2016年3月,采用焦点动物取样法和随机取样法对短须裂腹鱼的行为谱进行了研究。以"姿势、动作和环境"为轴心,以生态功能为分类依据,构建短须裂腹鱼PAE行为谱。共辨识和定义了自然条件下短须裂腹鱼的10种姿势、22种动作和43种行为。根据行为的生物学功能,将上述行为划分为11大类,即摄食与排遗、探索、领域、发情、交配、护幼、冲突、运动、聚群、休息和杂类行为;区别了各种行为在雄性、雌性和幼体之间的相对发生频次以及发生季节。本研究有助于短须裂腹鱼的行为与保护研究,亦可为其他鱼类的行为谱研究提供参考。     

短须裂腹鱼胚胎与仔鱼早期发育特性研究

为积累短须裂腹鱼的基础生物学资料,并对规模化人工繁殖提供理论指导,利用养殖条件下的短须裂腹鱼亲鱼,通过干法授精获得受精卵,对其胚胎发育和仔鱼早期发育全过程进行连续观察。短须裂腹鱼受精卵的平均卵径为2.36 mm,吸水膨胀后为3.68 mm,沉性卵、弱粘性、卵黄丰富。在水温(14±1)℃的条件下,受精后6 h 30 min进入卵裂期,20 h 55 min进入囊胚期,60 h 28 min进入原肠期,70 h 4 min进入原肠中期,77 h 52 min进入神经胚期,142 h肌肉开始收缩,177 h 46 min进入心动期,254 h 40 min开始出膜。孵化全过程所需积温为3 565.3℃·h。初孵仔鱼全长8.7 mm。出膜后第2~9天,胸鳍、鳃、口腔、眼色素、体内血管等器官相继发育完全;第10天仔鱼全长达15.15mm,鳔充气,鱼苗开始平游和觅食。孵化过程中应重点防控水霉病。早期仔鱼可以投喂轮虫、蛋黄或者豆浆等,待仔鱼捕食能力变强之后,可以投喂更加适口的枝角类、嫩口丰年虫等。     

短须裂腹鱼仔稚鱼发育及生长特性的初步研究

为了掌握长江上游特有鱼类短须裂腹鱼(Schizothorax wangchiachii)的基础生物学资料,遏制其自然资源迅速下降态势,维护长江鱼类的生物多样性,在水温10.0～20.4℃的条件下,对人工催产孵化的短须裂腹鱼仔稚鱼形态发育特征进行了连续观察,并逐日记录其生长情况,分析其生长特性。结果显示,刚出膜短须裂腹鱼仔鱼全长(9.83±0.88) mm,呈现透明淡黄色,卵黄囊较大,胸鳍较小,仔鱼多侧卧静栖水底,人为刺激下可以向前移动;23日龄仔鱼全长(15.18±0.52) mm,卵黄全部消失,鳔室呈长圆柱形,肠道内充满食物,进入稚鱼期;35日龄稚鱼全长(16.75±0.75)mm,出现二鳔室、腹鳍呈现月牙状,腹鳍褶皱宽大;65日龄稚鱼全长(23.64±0.82) mm,腹鳍褶皱几乎消失,除未见臀鳞外,各鳍均已出现,其生活习性已与成鱼相似。短须裂腹鱼仔稚鱼的生长呈现先急速增加、而后平缓、再快速增加、而后变慢的过程,特定生长率呈现先迅速增加然后指数型下降的趋势。运用Matlab对短须裂腹鱼体长(L_P)与日龄(t)的多项式关系函数(L_P=9.7296+0.55051t-0.0239t~2+5.1283×10~(-4)t~3-3.4419×10~(-6)t~4(R=0.9666,SD=0.6824,P0.001)进行分析,求得22.14日龄和52.35日龄是短须裂腹鱼仔稚鱼生长变化的关键日龄,与个体发育阶段的卵黄消失(23日龄)和52日龄左右(20 mm左右)稚鱼高死亡率接近。     

一株鲈鲤源鲤春病毒血症病毒的分离鉴定及其病理观察

2016年4月,四川某鲈鲤(Percocypris pingi)养殖场流行一种以鳃、鱼鳔和内脏器官出血为临床特征的传染病。组织病理学观察发现,患病鲈鲤全身多组织器官均发生明显的病理损伤,尤其是肝、脾、肾、鳃和肠表现为明显的出血、变性、坏死以及炎症细胞浸润。取病鱼组织匀浆滤液接种鲤上皮瘤细胞(epithelioma papulosum cyprini, EPC),盲传3代出现典型的细胞病变(cytopathic effects, CPE)。将自然发病鱼组织匀浆滤液和细胞培养病毒液分别接种健康鲈鲤,实验鱼出现与自然发病鱼相同的症状,死亡率分别为60%和50%,而对照组未见异常。对经分离毒株ZLP160415感染出现CPE的EPC细胞制备超薄切片进行电镜观察,发现病毒呈弹状,长90～150 nm,宽40～60 nm;对自然发病鱼、人工感染发病鱼内脏组织和细胞培养病毒液进行鲤春病毒血症病毒(springviremiaofcarpvirus,SVCV)的RT-PCR检测,均扩增出目的条带。基于SVCV糖蛋白基因进行系统发育分析,结果显示分离毒株(ZLP160415)属于Ia型。结合本次疾病的流行病学与病理损伤特点、病毒分离鉴定、人工感染试验结果和透射电镜检查,确定此次流行病的病原为SVCV。     

光照对短须裂腹鱼生长及生理影响研究

光照是影响鱼类活动、摄食和生长的主要环境因子之一。本文分别以野生和养殖短须裂腹鱼(Schizothorax wangchiachi)为研究对象,研究了短须裂腹鱼对光谱、光强选择趋向性及光谱对短须裂腹鱼生长、生理的影响。结果显示,黄光、红光和绿光都对短须裂腹鱼起到吸引作用;利用黄光进行光照强度选择性研究发现,在低光照强度范围(22.6~64.7 lx),短须裂腹鱼对光强的趋向性没有显著差异(P>0.05);为期56 d的养殖实验发现,不同的光照颜色显著影响短须裂腹鱼的生长率和存活率(P<0.05),黄光和自然光处理组的养殖效果无显著性差异;无论在生长率和应激水平,均优于蓝光环境,蓝光提升了短须裂腹鱼的应激水平,导致存活率下降。研究结果可为水利工程建设过鱼设施诱鱼光源的选择提供科学依据。     

魁蚶稚贝的底质选择性及其潜沙能力评价

底质是魁蚶稚贝潜沙、水平移动、存活与生长的重要环境因素,选择底质适宜的海区开展底播增殖,可以有效提高魁蚶稚贝的增殖效果,但目前仍缺乏相关研究。因此,在实验室条件下,研究了10 mm<壳长<15 mm、15 mm<壳长<20 mm、20 mm<壳长<25 mm、25 mm<壳长<30 mm 4种规格魁蚶稚贝对4种底质粒径(细砾、粗砂、中砂、细砂)的选择模式,并量化分析了20 mm<壳长<30 mm稚贝在5种底质粒径(增设泥底质)条件下的潜沙能力。结果表明:(1)10 mm<壳长<15 mm稚贝的足丝较发达,以附着生活为主,明显偏好粗砂底质,其在两种底质交界位置的分布率、水槽的贴壁率均最高;15 mm<壳长<30 mm稚贝转入营埋栖生活,随着规格的增大,稚贝选择的底质粒径亦增大,而在2种底质交界位置的分布率、水槽贴壁率也随之减小。其中15 mm<壳长<20 mm稚贝偏好选择中砂,分布率为50.50%,其次为细砂;20 mm<壳长<25 mm稚贝对不同粒径底质的适应性均较强;25 mm<壳长<30 mm稚贝偏好选择粗砂底质,分布率达到65%,其次为中砂。(2)20 mm<壳长<30 mm稚贝在淤泥底质中的初潜时间最短,为2.74 min,其次为中沙、细沙;在细砂底质中的潜沙率最高,达到85%,其次为中沙。综合分析认为,20 mm<壳长<30 mm稚贝是较适宜的底播苗种规格,而以中砂(0.25 mm<粒径<0.5 mm)和细砂(粒径<0.25 mm)为主要粒径组成的海区宜进行魁蚶稚贝的底播增殖。     

Effect of sand grain size on substrate preference and burial behaviour in cultured Japanese flounder juvenile,Paralichthys olivaceus

To improve release techniques of Japanese flounder, Paralichthys olivaceus, for the purpose of stock enhancement, the substrate preference and burial behaviour over a range of sand grain sizes were investigated under laboratory conditions. Ten 12‐hr trials (i.e. 10 experimental groups), which, respectively, contained two different levels of substrates, demonstrated that juvenile flounder (55–78 mm total length) selected substrate of grain size (GS) < 2.00 mm, and the degree (from high to low) of their preference could be listed as Level 3 (0.50–2.00 mm) > Level 4 (0.25–0.50 mm) > Level 5 (0.05–0.25 mm) > Level 2 (2.00–4.00 mm) > Level 1 (4.00–6.00 mm). The average burying of fish in substrates of GS < 2.00 mm was more than 93.1%. Additionally, along with food deprivation time increased from 12 to 24 hr, some flounder showed random off‐bottom swimming behaviour, and the appearance of off‐bottom swimming behaviour was <3.2%. These results suggest that releasing sites with appropriate substrate grain size (GS < 2.00 mm) are suitable for conducting mass releases of hatchery‐reared flounder juveniles, and starvation prior to release should be minimized. The results of the present study are essential to improve stocking strategies of Japanese flounder and reduce the juvenile mortality after release.     

Substrate selection by subyearling European river lampreys (Lampetra fluviatilis) and older larvae (Lampetra spp)

More information on the habitat requirements of lamprey is needed to improve measures aiming at rehabilitation of lamprey populations. Knowledge on habitat requirements of subyearling lampreys is especially scarce. We studied substrate selection by subyearling lamprey during the first three months after hatching using aquarium experiments. Differences in substrate selection by different‐sized larvae older than one year and the substrate depth selection by large larvae were also studied. Prolarvae of an average total length (TL) of 6.9 mm actively selected substrates with existing holes. When lampreys reached TL of over 8.0 mm, they started to select substrates which enabled them to construct a burrow. Lampreys of that size avoided coarse sand and gravel without fine material. Natural gravel with fine material was a frequently selected burrowing substrate during the 10‐week experiment suggesting its importance for subyearling lampreys. Substrates where very fine sand prevailed were the most selected soft sediments by subyearling larvae. Larvae older than one year did not select clay as a burrowing substrate. Medium (TL 70–98 mm) and large (TL 110–146 mm) larvae preferred coarse organic matter to fine sand. Larvae older than one year (TL 80–110 mm) preferred the thickest layer of soft substrate (10 cm) available to thinner layers.     

短须裂腹鱼有氧游泳能力及其行为的实验研究

研究短须裂腹鱼的游泳能力、运动生理及游泳行为,为过鱼设施的设计提供参考依据。实验用15尾短须裂腹鱼于2013年7月中旬采自金沙江上游玛曲河河口处,体长(23.83±2.47)cm,体重(224.95±76.83)g。游泳能力测试装置采用丹麦Loligo Systems公司生产的大型游泳水槽。(1)短须裂腹鱼临界游泳速度为(75.04±7.6)cm/s、(3.17±0.42)BL/s;(2)运动耗氧率与游泳速度呈幂函数关系:MO2=100.00+42.61U1.81(R2=0.995,P0.001);单位距离耗氧率(COT)与游泳速度的关系也呈幂函数关系:COT=0.12U-1+0.04U1.02(R2=0.898,P0.001),最适游速Uopt=1.81 BL/s,COTmin=0.14 mg/(kg·m);(3)随着游泳速度的增加,尾摆幅度的变化不显著(P0.05),变化范围为0.17~0.26 BL、平均(0.21±0.02)BL,而尾摆频率和运动步长都呈线性增加的趋势。     

Positive phototaxis as the cause of jaw malformations in larval greater amberjack,Seriola dumerili (Risso, 1810): Mitigation by rearing in tanks with low‐brightness walls

Jaw malformations are a serious problem in fingerlings of the greater amberjack, Seriola dumerili. To establish a countermeasure against these malformations, we conducted a larval behavioural test to identify their root cause, and we investigated the ability of low‐brightness rearing tank walls to control their occurrence. Larval distribution was examined in a horizontally illuminated rectangular transparent aquarium (20 cm × 100 cm × 25 cm), and their distribution was biased towards the light source after this lighting was provided from 3 to 13 days post hatching (dph), indicating positive phototaxis; this response disappeared on 22 dph (there are no data between 13 and 22 dph). Larvae were observed to swim phototactically towards the light source and repeatedly collided with the tank wall (walling behaviour). To investigate the ability of wall brightness to control the incidence of jaw malformations, we compared their incidence in a rearing trial with different Munsell colour values for the tank walls: 2.11 (black), 5.85 (grey) and 9.52 (white). The incidence rate of jaw malformations at 25 dph was significantly lower in the tanks with lower‐brightness walls, that is, 5.8%, 22.5% and 26.8% for black, grey and white respectively. Larval walling behaviour was most intense for white, followed in order by grey and black. These results indicate that positive phototaxis elicits larval walling behaviour, causing jaw malformations, and that low‐brightness tank walls control this process.     

Intra‐annual changes in reproductive indices of male and female Himalayan snow trout,Schizothorax richardsonii (Gray, 1832)

Periodic changes in reproductive hormone levels, gonadal histology and gonadosomatic index (GSI) of snow trout, Schizothorax richardsonii, were examined to ascertain annual cycle of gonadal development and reproductive status in their natural habitat. In females, there were coherent changes in plasma 17β‐oestradiol and vitellogenin along with GSI, oocyte maturation and vitellogenic progression, collectively indicating two distinct maturation peaks during the months of September and February. Coinciding with this, in males, plasma 11‐keto testosterone was also noticeably higher during September and February, with highest GSI values in September. However, plasma 17α, 20β‐dihydroxyprogesterone levels in males were found to be persistently high from September to February. This observation suggests the potential presence of matured oozing males over a longer period, unlike in females. Overall, the close association between reproductive hormone levels, GSI and gonadal maturation stages in males and females (particularly, the presence of postovulatory follicle complexes) with apparent natural synchronization clearly indicates that S. richardsonii breeds twice in a year, possibly during late September to early November and late February to early April in the coldwater riverine habitats of the Indian Himalayan region.     

Activities of digestive enzymes in relation to ingested natural food in three carp species of the Himalayan River Ladhiya,northern India

The present investigation was carried out to study gut content and accompanying digestive enzyme profiles of the three carp species, golden mahseer Tor putitora, snow trout Schizothorax richardsonii and hill trout Barilius barila. The fishes were captured from the River Ladhiya, Champawat district, Uttarakhand, northern India. The digestive tract length was on average 1.8 and 2.3 times longer than the body, while members of Bacillariophyceae, Chlorophyceae, Cyanophyceae and Zygnematophyceae made up 95.15% and 99.63% of gut content in the golden mahseer and snow trout, respectively. In hill trout, the digestive tract being only half the body length, gut content was insects (72.73%), eggs of aquatic organisms (22.73%) and nematodes (4.55%). Estimated gut enzyme activities demonstrated that amylase activity was significantly (P < 0.05) higher in golden mahseer and snow trout compared to hill trout, while total protease, trypsin and chymotrypsin activities were all significantly (P < 0.05) higher in the hill trout compared to other species. A strong co‐relation was found among relative gut length, food composition and digestive enzyme activities of each species. This may guide the future formulation of feed to these important species of carps kept in farm.     

Construction and evaluation of type III secretion system mutants of the catfish pathogen Edwardsiella piscicida

Catfish is the largest aquaculture industry in the United States. Edwardsiellosis is considered one of the most significant problems affecting this industry. Edwardsiella piscicida is a newly described species within the genus Edwardsiella, and it was previously classified as Edwardsiella tarda. It causes gastrointestinal septicaemia, primarily in summer months, in farmed channel catfish in the south‐eastern United States. In the current study, we adapted gene deletion methods used for Edwardsiella to E. piscicida strain C07‐087, which was isolated from a disease outbreak in a catfish production pond. Four genes encoding structural proteins in the type III secretion system (T3SS) apparatus of E. piscicida were deleted by homologous recombination and allelic exchange to produce in‐frame deletion mutants (EpΔssaV, EpΔesaM, EpΔyscR and EpΔescT). The mutants were phenotypically characterized, and virulence and vaccine efficacy were evaluated. Three of the mutants, EpΔssaV, EpΔyscR and EpΔesaM, were significantly attenuated compared to the parent strain (p < .05), but EpΔescT strain was not. Vaccination of catfish with the four mutant strains (EpΔssaV, EpΔesaM, EpΔyscR and EpΔescT) provided significant protection when subsequently challenged with wild‐type strain. In conclusion, we report methods for gene deletion in E. piscicida and development of vaccine candidates derived from a virulent catfish isolate.     

Periphyton characteristics influence the growth and survival of Holothuria scabra early juveniles in an ocean nursery system

Floating hapas (fine mesh net enclosures) are a cost‐effective ocean nursery system to culture post‐metamorphic Holothuria scabra to release size. The growth of periphyton biofilm on hapas is a natural food source for early juveniles. This study investigated the effects of periphyton quality (i.e. chlorophyll‐a, phaeopigment, total biomass, autotrophic index or AI), water quality (nutrients, chlorophyll‐a) and environmental parameters (temperature, rainfall) on the temporal variation in the growth and survival of early juvenile (~3 mm) H. scabra reared in floating hapas. Five trials where the juveniles were reared for 60 days each in the eutrophic coastal waters of Bolinao, the Philippines were conducted during different months over 2 years. Significant differences in the growth and survival of juveniles among trials were found. Absolute growth rates (AGR) ranged widely (0.01–0.09 g/day). Trials with high AGR of juveniles (0.07–0.09 g/day) during the first 30 days of rearing had significantly higher chlorophyll‐a (chl‐a) in biofilm (15.9–27.5 mg/m²) and lower AI. Conversely, during the subsequent 30 days, trials with high AGR of juveniles (0.06–0.11 g/day) had significantly lower chl‐a and higher AI. Multivariate analyses showed that chl‐a in biofilm, AI and nutrients in the water column are good indicators of periphyton quality and juvenile growth rates in floating hapas. Further, this study validates the expansion of the feeding mode of juveniles from primarily grazing on microalgae, to feeding on detritus and heterotrophs as they grow. These results are important in optimizing ocean nursery systems.