中华倒刺鲃仔、稚鱼的耳石微结构与日轮形成特征

以中华倒刺鲃(Spinibarbus sinensis)亲鱼为材料,在实验室养殖条件下以人工孵化的丰年虫(Eubranchipus vernalis)为仔、稚鱼饵料,通过已知日龄法观察其耳石的微结构,分析其日轮形成特征。结果表明,仔、稚鱼的微耳石和矢耳石一般由1个中心核和1个耳石原基组成,少数存在多个中心核或原基现象,星耳石中心核和原基区分不明显。微耳石和矢耳石中心核直径分别为(37.73±5.34)μm和(39.78±7.11)μm,耳石原基直径分别为(16.29±3.46)μm和(17.09±3.88)μm。矢耳石和星耳石轮纹清晰度、规律性、周期性和完整性不及微耳石;微耳石第1条日轮在仔鱼出膜后第2天形成,以后每天沉积1轮。30日龄稚鱼微耳石轮纹数(N)与日龄(T)的关系符合直线模型,相关关系式为:N=1.0016T-0.8753(R2=0.9961,P<0.01,n=197),线性方程斜率与1无显著性差异(P>0.05)。在微耳石和矢耳石样本中共观察到孵化标记轮和转移标记轮2种,其中孵化标记轮的出现率分别为78.68%和83.33%,转移标记轮的出现率分别为29.95%和48.98%。60尾33日龄稚鱼微耳石的生长轮宽度变化范围0.522~2.244μm,平均为(1.087±0.231)μm。     

叉尾斗鱼仔鱼耳石形态发育与日轮形成特征

对实验室人工繁殖的叉尾斗鱼(Macropodus opercularis)仔鱼耳石形态发育与日轮进行了观察研究.结果显示,叉尾斗鱼微耳石和矢耳石在胚胎时已出现.微耳石在仔鱼刚孵化时为圆盘状,随仔鱼发育转变为近椭圆形,孵出后19 d转变为中部圆凸两端较尖的菱形;矢耳石在仔鱼刚孵出时为圆盘状,随仔鱼发育转变为椭圆形,孵出后19 d转变为一端略尖的桃形;星耳石在仔鱼孵出后第19天才出现,呈中部略凹的椭圆形.叉尾斗鱼仔鱼耳石长径(包括微耳石和矢耳石)与鱼体全长(TL,mm)呈线性相关.仔鱼耳石上第一个轮纹在孵出后第2天形成,其后每日形成1个新轮纹,生长轮数目与仔鱼日龄(D)呈线性相关,且线性方程斜率接近于1.结论认为,叉尾斗鱼仔鱼星耳石出现时间晚,矢耳石形态在后期出现较大变化,而微耳石在胚胎时即形成,形态稳定;日轮可读性较好,故更适合做日轮研究的材料.本研究旨在为叉尾斗鱼自然种群年龄结构调查及其资源保护提供基础数据.     

东海条石鲷仔鱼耳石日轮与生长的关系

2009年4月15日-5月5日在浙江省舟山水产研究所条石鲷(Oplegnathus fasciatus)人工繁殖期间,逐日选择胚胎和仔鱼样本,连续解剖观察发育后期胚胎和前期仔鱼,光镜观察仔鱼矢耳石和微耳石的形态、日轮数,测定其直径,研究条石鲷的耳石日轮和生长。结果表明,受精后约26h,条石鲷胚胎听囊内出现1对矢耳石和1对微耳石;仔鱼孵出第2天形成第1个轮纹,之后每天形成1轮,孵化后天数(N)和矢耳石日轮数(D)的关系为N=D+1;在第8天左右,矢耳石上出现第2条明显的标记轮,为初次摄食轮。仔鱼耳石长径(rs,μm)与鱼体体长(L,mm)呈线性相关,其关系式为rs=18.146L-44.436;矢耳石长径rs与微耳石长径rl之间存在线性相关,其关系式为rs=0.6125rl+1.9882。根据结果确认,矢耳石轮纹可作为条石鲷仔鱼日龄的判别依据。     

长江口凤鲚仔稚鱼不同发育阶段矢耳石生长

2014年4—8月于凤鲚(Coilia mystus)的主要繁殖期在长江口采集仔稚鱼103尾。分析凤鲚仔稚鱼样本得出日龄范围为5~48 d,体长为4.20~26.21 mm。为研究长江口凤鲚早期发育不同阶段生长,测定了凤鲚仔稚鱼(5~48日龄)体长随日龄的生长变化,其变化分为3个阶段(5~11日龄、12~30日龄、31~48日龄)。不同日龄阶段,体长的生长速率差异性显著(P0.05)。耳石早期发育研究发现耳石长、耳石宽随鱼体的生长而增长,在17~19日龄之后其生长速率增大约2倍。采用分段回归方法分析耳石长、耳石宽与日龄的关系,发现二者异速生长的拐点均出现在19~20日龄,拐点前为慢速生长,之后为快速生长。研究发现凤鲚早期生长发育阶段的耳石形态有较为显著的改变:卵黄囊期、前弯曲期为圆形;弯曲期耳石长的生长大于耳石宽的生长,耳石逐渐变成椭圆形;耳石变成稳定形态后,长宽比基本保持稳定。     

大黄鱼仔稚鱼不同发育阶段矢耳石形态发育和微结构特征

对人工培育大黄鱼(Larimichthys crocea)的生长发育与矢耳石形态及微结构特征进行研究,结果表明:(1)大黄鱼矢耳石上的轮纹是每日形成的,第1日轮在孵化后第2天形成,与其初次摄食相对应。(2)大黄鱼卵黄囊期和前弯曲期仔鱼的耳石形态为圆形,进入弯曲期耳石长轴迅速伸长,在后弯曲期耳石形态变为椭圆形。进入稚鱼期,矢耳石开始形成次生核。随后次生核数量逐渐增加,在孵化后47～78 d的个体中,次生核数量稳定在5～7个,耳石近似盾形。(3)根据耳石日轮宽度推算的大黄鱼稚鱼在其仔鱼期生长率(b)与第1个次生核的形成时间(tsp1)之间存在明显的线性关系,表明生长较快的个体形成次生核的时间较早,进入稚鱼期所需的时间更短。以上结论表明,大黄鱼矢耳石可以反演其早期生活史阶段的生长发育特征。     

黑脊倒刺鲃仔、稚、幼鱼的生长发育研究

黑脊倒刺鲃的仔、稚、幼鱼发育的特点基本上和四大家鱼一致,仅卵黄囊消失得较迟。黑脊倒刺鲃从1日龄早期仔鱼发育到32日龄早期幼鱼期间,体长平均日增长0.4 mm,体重平均日增长5.65 mg。体长增长呈直线增长,体重和体长关系呈幂函数关系。     

中华倒刺鲃仔稚鱼期消化酶及碱性磷酸酶活性变化的研究

本试验选取了中华倒刺鲃从初孵仔鱼到30日龄稚鱼,分析测定了中华倒刺鲃的特定生长率、胰蛋白酶、淀粉酶、脂肪酶和碱性磷酸酶的活性。结果表明：在整个试验期间,中华倒刺鲃的SGR为26.43%。在初孵仔鱼体内就能检测到胰蛋白酶、淀粉酶、脂肪酶和碱性磷酸酶的活性。胰蛋白酶在内源营养阶段具有较高的活性,在4日龄开口时其比活力降到一较低值,随后呈现增加的趋势,在18日龄达到最低值后又缓慢增加。淀粉酶在试验期间表现出较高的活性,比活力一直呈增加的趋势。脂肪酶在18日龄前其比活力一直增加,并且在7-18日龄表现出较高的比活力水平,18日龄后快速下降。碱性磷酸酶活性则呈现出一直增加的趋势。     

倒刺鲃属3种鱼类种间杂交的初步研究

采用人工催产和干法授精技术,进行了以中华倒刺鲃、倒刺鲃为母本与其父本及黑脊倒刺鲃雄鱼的种间杂交实验,获得4个杂交组合和2个自交组合,在(27±0.5)℃温度条件下,对其F1卵膜径、受精率、孵化率、孵化时间、畸形率、子代早期形态特征与成活率进行了比较,同时,观察了胚胎发育及其异常现象,结果表明:以中华倒刺鲃为母本的F1卵膜径差异不显著;以倒刺鲃为母本的F1卵膜径,在多细胞期时差异不显著,在耳石期时,倒刺鲃♀×中华倒刺鲃♂F1卵膜径显著大于母本(P<0.05)。杂种F1的受精率、孵化时间与母本差异不显著,畸形率显著高于母本(P<0.05)。中华倒刺鲃杂种F1的孵化率较高,与母本差异不显著(P<0.05)。倒刺鲃♀×中华倒刺鲃♂F1的孵化率与母本之间差异不显著,倒刺鲃♀×黑脊倒刺鲃♂F1的孵化率显著低于母本(P<0.05)。中华倒刺鲃杂种F1仔鱼早期发育阶段存在死亡高峰期,成活率显著低于母本(P<0.05)。中华倒刺鲃种间杂交和倒刺鲃♀×黑脊倒刺鲃♂杂种F1既具有父本特征,同时又具有母本特征,初步证明倒刺鲃属种间杂交是可行的。     

三门湾和吕四近岸小黄鱼仔稚鱼耳石微结构特征比较

对三门湾和吕四近岸小黄鱼(Larimichthys polyactis)仔稚鱼的矢耳石微结构特征进行了比较研究。结果表明:小黄鱼矢耳石上的轮纹是每日形成的,第一日轮在孵化后第2天形成,由仔鱼期向稚鱼期过渡时形成的次生核;三门湾和吕四近岸海域在个体的生长、发育和次生核之间均表现出明显的一致性:生长较快的个体,第一次生核形成的时间也较早,次生核数量亦多;两个海域小黄鱼的耳石微结构存在明显的差异,吕四近岸的个体在仔鱼期耳石生长率更快(均值为0.039),第一次生核形成时间较短(均值为37日龄),次生核数量较多(7⁸个),表明吕四近岸个体具有较快的生长率和发育速度,这种差异可能是因为吕四近岸的个体在仔鱼期所经历的水温较高。以上研究结果表明,矢耳石的微结构特征可以反映野生小黄鱼的早期生长发育状况,不同海区小黄鱼仔稚鱼的矢耳石微结构特征存在显著差异,可以用于区分小黄鱼不同海域栖息的个体,将为小黄鱼早期资源补充过程和早期选择性死亡等研究提供理论基础。     

半刺厚唇鱼胚胎与胚后发育观察

通过对半刺厚唇鱼[Acrossocheilus(Lissochilichthys)hemispinus]胚胎及胚后发育进行观察,详细描述了从受精卵到稚鱼期发育时期的时序和形态特征.结果表明:半刺厚唇鱼成熟卵圆形,卵径1.74～1.97 mm,黄色或桔黄色.在25 ～28℃水温条件下,孵化时间为62 h,刚出膜仔鱼全长5.6～7.2 mm.在26～28℃水温条件下,出膜后仔鱼7日龄初次摄食;9日龄时卵黄囊完全消失;15日龄时,进入稚鱼期:25日龄时,鳞片形成,进入幼鱼期.仔、稚鱼体侧都具有7条明显的黑褐色横向条纹,成鱼条纹消失.     

Rare serotype occurrence and PFGE genotypic diversity of Streptococcus agalactiae isolated from tilapia in China

Previously, we reported 10 PEGE types of 85 tilapia Streptococcus agalactiae(GBS), which shifted from Streptococcus iniae in China, by using PEGE method. Presently, larger and more representative tilapia GBS were isolated, for the ?rst time in China, to characterize their serotypes and genetic diversities more precisely than had done before. 168 GBS strains were distributed in ?ve provinces of China, in which Guangdong, Guangxi and Hainan were the major ones, holding36.9%(62/168), 37.5%(63/168) and 19.6%(33/168), respectively. Serotypes, Ia, Ib and III, were observed in these strains and the most predominant one was Ia(95.2%), which mainly distributed in Guangdong, Guangxi and Hainan. Ia initially occurred in 2009, it shoot up to 32.1% in 2010,but decreased to 16.1% in 2011 before went up to 45.2% in 2012. Ib sporadically occurred during2007–2011, III onlyoccurred in 2012. 14 different PFGE types, including 4 new types(N, O,P and Q), were observed, in which B, D, F and G were the predominant types, holding 83.9%(141/168) of the total GBS strains. Ia corresponded to 11 PFGE types(A–H, N–P), in which type D predominated(51%). Ib represented 3 genotypes(I, J and Q) and III harbored only 2genotypes(N and F). Type N and Fsynchronously presented in Ia and III. In summary, the genetic diversity of tilapia GBS varied by serotypes and changed with geographical locations and years.Although Iastillpredominated, new rareserotypeIII alreadyoccurred in China.     

Growth hormone (GH) and reproduction: a review

Interaction between growth and reproduction occurs in many vertebrates and is particularly obvious at certain stages of the life cycle in fish. Endocrine interactions between the gonadotropic axis and the somatotropic axis are described, the potential role of GH being emphasised. A comparative analysis of these phenomena in mammals, amphibians and fish, suggests a specific role of GH in the physiology of puberty, gametogenesis and fertility. It also shows the original contribution made by studies on the fish model in this field of investigations.     

Purification and partial characterization of GtHs (cfLH and cfFSH) from Indian walking catfish (Clarias batrachus) (L.) and development of a homologous ELISA for cfLH

Two gonadotropins (GtH; Qa and Qb) were purified by gel filtration and ion exchange chromatography from the pituitaries of Indian walking catfish (Clarias batrachus). The presence of GtH during purification was assessed by in vitro oocyte maturation and in vivo steroidogenic activity, and their identities were determined by elution profiles, molecular weight, biological activities and yield. The molecular weights of Qa and Qb were 37 and 42 kDa, respectively, and composed of distinct subunits (Qa: 20 and 14 kDa and Qb: 26 and 18 kDa). Polyclonal antibodies raised against Qa immunostained Qa, Qb and pituitary GtH cells. A competitive Qa‐ELISA was developed whose sensitivity was 6.25 ng mL^?1 (1.25 ng well^?1) with intra‐ (3.5%) and inter‐ (12.4%) assay coefficients of variation. Displacement curves parallel to the standard were obtained with plasma and pituitary extracts of catfish, Qb and carp GtHII. The assay was validated by measuring the plasma Qa levels after LHRH treatment and in relation to ovarian growth in the female catfish during different reproductive phases. Based on the results, Qa and Qb corresponded to fish LH and FSH respectively. The findings will increase the knowledge of the mechanisms controlling fish reproduction and identification of sensitive phases in fish in captivity for hormonal manipulation.     

Controlled infection of Poecilia reticulata Peters (guppy) with Tetrahymena by immersion and intraperitoneal injection

Tetrahymena is a protozoan parasite, which infects guppy, Poecilia reticulata Peters, and causes substantial economical losses in commercial farms worldwide. Studies of guppy infected by Tetrahymena require standardized infection protocols. The LD50 for Tetrahymena infection of guppies by intraperitoneal (IP) injection was calibrated, and the level obtained was 946 parasites per fish. Guppy infection with Tetrahymena by immersion, imitating the natural route of infection via the integument, was studied under normal or stress conditions. Exposure to cold and netting (CNI) and to cold only (CI) followed by immersion exposure to 10 000 Tetrahymena per mL resulted in 22.5% and 19.2% mortality, respectively, as compared to 14.2% and 10% in groups that were netted only (NI) or non‐stressed (I). Histopathology revealed that immersion infection resulted in a systemic infection. Lysozyme levels, measured 3 weeks after infection, were significantly higher in the CNI group (288 μg per mg protein) compared with CI‐, NI‐ and I‐treated groups (94.5, 64 and 62.3 μg mg^?1, respectively). There was no evident parasite immobilization activity in body homogenates, suggesting no development of acquired immunity. Re‐infection by IP injection revealed no increase in protection in any of the treatment groups, mortality range of 56.3–75%, higher than in the non‐exposed control (40.6% mortality).     

Evaluation of a Fast Method Based on the Presence of Two Restriction Sites in the Mitochondrial ND5 (mt ND5) Gene for the Identification of Scomber Species

The purpose of this work was to evaluate the suitability of a method based on the presence of two restriction sites (for Hae III and Hindf I) in the mitochondrial NADH dehydrogenase subunit 5 (mt ND5) gene to identify Scomber species. The evaluation was performed on 144 reference and market samples by sequencing of the entire 505-bp fragment of the mt ND5 gene and of a 464-bp fragment of the Kocher fragment of the cytochrome b gene (mt Cytb). Sequence analysis of any of the two fragments allows the identification of each of the four Scomber species, but S. japonicus and S. colias had the same restriction sites at the ND5 amplicon and would not have been differentiated by this analysis. Similarly, loss of the Hae III site in some S. scombrus individuals would have misidentified them as not being Scomber. All the market products were correctly labeled except one acquired in Spain labeled as originating in the Atlantic and containing S. japonicus.     

Nitrogenous waste excretion and accumulation of urea and ammonia inChalcalburnus tarichi (Cyprinidae), endemic to the extremely alkaline Lake Van (Eastern Turkey)

The endemic, anadromous cyprinidChalcalburnus tarichi is the only fish species known to occur in alkaline Lake Van (Eastern Anatolia, Turkey). EightC. tarichi were maintained individually in Lake Van water (17 – 19°C; pH 9.8; 153 mEq·I^–1 total alkalinity; 22 total salinity) and tank water samples analyzed for 24 h in 2 to 4 h intervals. At zero time, < 1µM ammonia was present and urea was undetectable in the tank water; at 24 h, total ammonia and urea made up 114±32 and 35±25µM, respectively. Over the experimental period, ammonia-N and urea-N excretion averaged 1041±494 and 607±169moles·kg^–1 fish·h^–1, respectively. The extent of urea excretion was highly variable between specimens. Uric acid excretion was not detectable.Urea was present at high concentrations in all tissues and plasma (25 – 35moles·g^–1·ml^–1) of freshly caughtC. tarichi; total ammonia content of the tissues was by a factor of 1.9 (liver) to 3.0 (brain) lower. High arginase activity (2.4±0.2 U·min^–1·g^–1) was detected in the liver ofC. tarichi but ornithine carbamoylphosphate transferase, a key enzyme of the ornithine-urea-cycle, was absent. Ureagenesis is likely through degradation of arginine and/or uricolysis. High glutamine synthetase activity (11±0.6 U·min^–1·g^–1) and low ammonia content in brain suggest that, like other teleosts,C. tarichi has an efficient ammonia detoxification in the brain, but in no other tissue.Nitrogenous waste excretion at alkaline pH is discussed. The ability ofC. tarichi to excrete high levels of ammonia at extremely alkaline pH is unique among teleosts studied so far. The mechanism of ammonia excretion under Lake Van conditions remains to be elucidated.     

Imported ornamental fish are colonized with antibiotic‐resistant bacteria

There has been growing concern about the overuse of antibiotics in the ornamental fish industry and its possible effect on the increasing drug resistance in both commensal and pathogenic organisms in these fish. The aim of this study was to carry out an assessment of the diversity of bacteria, including pathogens, in ornamental fish species imported into North America and to assess their antibiotic resistance. Kidney samples were collected from 32 freshwater ornamental fish of various species, which arrived to an importing facility in Portland, Oregon from Colombia, Singapore and Florida. Sixty‐four unique bacterial colonies were isolated and identified by PCR using bacterial 16S primers and DNA sequencing. Multiple isolates were identified as bacteria with potential to cause disease in both fish and humans. The antibiotic resistance profile of each isolate was performed for nine different antibiotics. Among them, cefotaxime (16% resistance among isolates) was the antibiotic associated with more activity, while the least active was tetracycline (77% resistant). Knowing information about the diversity of bacteria in imported ornamental fish, as well as the resistance profiles for the bacteria will be useful in more effectively treating clinical infected fish, and also potential zoonoses in the future.     

Effect of iodophor disinfection of non‐hardened Salmo trutta eggs on their bacterial and fungus load

This study investigated the efficiency of iodophor disinfection (135 ppm active iodine for 15–30 min) of non‐hardened Salmo trutta eggs against different groups of bacteria and against fungus. Egg samples were taken from non‐disinfected and from disinfected eggs, microorganisms were cultured on specific nutrient media and their mass was measured by turbidimetric methods. Bacteria and fungus mass of non‐hardened eggs could be reduced but not eliminated by iodophor disinfection with 135 ppm active iodine for 15 min. The extent of reduction was 47–65% (Experiment 1). The efficiency of disinfection increased with disinfection time as the reduction in bacteria and fungus mass was 40–55% after 15 min and 58–74% after 30 min (Experiment 2). Disinfection efficiency of iodophor solution diluted in water (reduction 49–57%) and of iodophor solution diluted in sodium chloride solution iso‐osmolar to the oocytes (reduction 52–61%) was similar (Experiment 3). The reduction in bacteria and fungus mass was persistent as it was 39–72% lower in embryos deriving from disinfected eggs than in embryos deriving from non‐disinfected ones (Experiment 4). In conclusion, the tested disinfection method is inadequate to eliminate pathogens completely but it could positively influence immune defence of eggs and embryos.