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1.
The purpose of this study was to compare the performance of two species of diatoms, Thalassiosira weissflogii and Chaetoceros gracilis, in the larviculture of the black tiger shrimp Penaeus monodon. Shrimp larvae were fed with either C. gracilis, T. weissflogii, or a combination of the two species of diatoms. The larvae fed solely with T. weissflogii or a combination of the two types of diatom had significantly higher survival rates and faster metamorphosis than those fed solely with C. gracilis. The numbers of diatom cells consumed by larvae during 3-h periods were determined, revealing that larvae consumed significantly higher numbers of C. gracilis than T. weissflogii. However, when the protein, total fatty acid, and polyunsaturated fatty acid content of the two species of diatom are compared, significantly higher amounts of each are found in T. weissflogii. Converting the number of diatom cells consumed into equivalent protein, total fatty acid, eicosopentaenoic acid, and decosahexaenoic acid reveals that larvae fed with T. weissflogii received significantly higher amounts of all the nutrients, compared to those consuming C. gracilis. The results showed an advantage of feeding T. weissflogii to C. gracilis in enhancing survival and metamorphosis in P. monodon larvae.  相似文献   

2.
To select a reliable and sensitive method for discriminating strains of Porphyra haitanensis, the nucleotide sequence of the internal transcribed spacer 1 to internal transcribed spacer 2 regions (ITS-5.8S) of nuclear ribosomal DNA and the intergenic spacer region of RUBISCO were compared in five wild and five cultivated Porphyra haitanensis strains. Based on molecular analyses, sequences of ITS-5.8S (about 1,210 bp) could be divided into three regions: ITS1, 5.8S, and ITS2. The ITS1 and ITS2 sequences of each strain differed, even between individuals collected from the same site. In contrast, 5.8S rDNA and RUBISCO spacer sequences were identical among the ten P. haitanensis strains, although differences were found among different Porphyra species. Phylogenetic analysis also supported these conclusions. These sequence features of highly conserved regions and diversified regions that occurred repeatedly in ITS-5.8S could be useful in discriminating germplasm of P. haitanensis strains or Porphyra species. In contrast, the RUBISCO spacer is only suitable for identifying Porphyra species. New coupled primers were designed to amplify only the 5.8S rDNA and ITS2 region of Porphyra. The sequences of these amplified fragments can be readily used to identify germplasm or to perform phylogenetic analysis of Porphyra spp.  相似文献   

3.
The significance of Mn and Fe for the growth of a coastal marine diatom Thalassiosira weissflogii was investigated by performing culture experiments containing macronutrients with either Mn or Fe, or both. Only the addition of both Mn and Fe induced the highest growth rates and maximal cell yields. Maximal growth was maintained in continuous culture media, which were repeatedly prepared by an inoculation of pre-culture and the addition of both Mn and Fe to the control culture medium containing macronutrients. In particular, it was found that the full growth recovery in Mn-sufficient medium (without added Fe) is accomplished by the addition of Fe even after several days’ incubation. On the contrary, there was no sufficient growth recovery by the addition of Mn after a long incubation time in Fe-sufficient medium but without additional Mn. These results suggest that T. weissflogii in Mn-sufficient waters retains the ability for full physiological recovery for a long time, probably resulting from the decrease in the oxidative stress of phytoplankton by the production of antioxidant enzyme Mn superoxide dismutase during a long incubation period.  相似文献   

4.
珠母贝属6个种的ITS 2分子标记研究   总被引:6,自引:3,他引:6  
对珠母贝属的大珠母贝、珠母贝、白珠母贝、黑珠母贝、长耳珠母贝、黑珠母贝和合浦珠母贝6个种的内部转录间隔区2(ITS2)序列及其两侧的5.8S和28S的部分序列进行了比较分析。其中黑珠母贝的序列来自GenBank。PCR扩增片段大小为600bp左右,测序结果表明,ITS2长211~254bp,两端的5.8S和28S分别长84bp和272bp(均含引物)。序列比对分析结果表明,5.8S和28S序列高度保守,不适合于种类鉴定,而ITS2序列高度变异,270个比对位点中有146个位点发生突变,其中72个位点发生插入/缺失突变。除白珠母贝和黑珠母贝之间的遗传距离较小外,其余种类之间的遗传距离远远大于种内遗传距离。基因型分析表明,每个种具有各自特有的基因型。基因型和序列变异分析表明ITS2序列可作为珍珠贝种类鉴定的分子标记。可用于种间、杂交育种、幼体和珍珠贝肉等材料的种类鉴定与遗传分析。  相似文献   

5.
Global distribution of platyhelminth parasites and their host specificities are not well known. Our hypothesis was that platyhelminth parasites of large pelagic fishes are common around the world. We analysed molecular variation in three different taxa of platyhelminth parasites infecting four species of tunas: yellowfin tuna (Thunnus albacares, Scombridae) from Western Australia, southern bluefin tuna (Thunnus maccoyii, Scombridae) from South Australia, Pacific bluefin tuna (Thunnus orientalis, Scombridae) from Pacific Mexico and northern bluefin tuna (T. thynnus, Scombridae) from two localities in the Mediterranean (Spain and Croatia). Comparisons of ITS2 and partial 28S rDNA demonstrated two congeneric species of blood flukes (Digenea: Sanguinicolidae) from multiple hosts and localities: Cardicola forsteri from southern bluefin and northern bluefin tunas, and Cardicola sp. from Pacific bluefin and northern bluefin tunas; and a gill fluke, Hexostoma thynni (Polyopisthocotylea: Hexostomatidae), from yellowfin, southern bluefin and northern bluefin tunas. Partial 28S rDNA indicates that a second type of fluke on the gills, Capsala sp. (Monopisthocotylea: Capsalidae), occurs on both southern bluefin and Pacific bluefin tunas. This appears to be the first report of conspecific platyhelminth parasites of teleosts with a wide‐ranging geographical distribution that has been confirmed through molecular approaches. Given the brevity of the free‐living larval stage of both taxa of flukes on the gills (H. thynni and Capsala sp.), we conclude that the only feasible hypothesis for the cosmopolitan distribution of these flatworms is migrations of host tunas. Host migration also seems likely to be responsible for the widespread occurrence of the two species of blood flukes (Cardicola spp.), although it is also possible that these were translocated recently by the spread of infected intermediate hosts.  相似文献   

6.
ABSTRACT:   Four species of Chattonella , which are well known to form red tides that are lethal to fish, were subjected to phylogenetic analysis on the basis of the ribosomal RNA genes (rDNA), 5.8S rDNA, 18S rDNA, 28S rDNA, and the flanking internal transcribed spacers 1 and 2 (ITS1 and ITS2). The 18S rDNA sequences of C. antiqua , C. marina , and C. ovata isolated from different regions in Japan were compared. They were found to be identical with each other in a sequence 1818 bp long. The sequences of the D1/D2 region in the 28S rDNA, 5.8S rDNA, and ITS region that are known to be more variable regions were also found to be identical. These homogeneities of the rRNA gene family revealed the extremely close relatedness of C. antiqua , C. marina , and C. ovata . The sequences of C. verruculosa were different from those of these three species , resulting in an 89.2% homology in the 18S rDNA sequences, 70.4% homology in the D1/D2 region in the 28S rDNA sequences, and an 81.5% homology in 5.8S rDNA sequences and the ITS regions. Chattonella verruculosa was grouped within a single cluster composed of Dictyochophyceae rather than the other species of Raphidophyceae.  相似文献   

7.
The significance of Mn and Fe for the growth of the coastal marine diatom Thalassiosira weissflogii was investigated by culture experiments in the presence of precipitated Fe(III) hydroxide [am-Fe(III)] and EDTA-Fe(III) complex with or without Mn addition. The culture experiments in all media without any added Mn(II) resulted in the very low phytoplankton growth for cell density and chlorophyll a (Chl a) concentration. In contrast, sufficient Mn addition (25 nM) induced the maximum growth both for cell generation and Chl a production. By using an approach in which further iron uptake by T. weissflogii from external iron in the culture media is prevented by adding hydroxamate siderophore desferrioxamine B (DFB) during cultivation, we examined the ability of T. weissflogii to grow on intracellularly stored Fe after the DFB addition. The addition of DFB after 3- and 5-days of cultivation resulted in the lower growth rate and lower maximum yields for cell density and Chl a concentration in solid 7-day-aged am-Fe(III) medium than in freshly precipitated am-Fe(III) medium. The longer aging time of am-Fe(III) in medium reduced the supply of bioavailable iron in the medium by the slower dissolution rate of am-Fe(III) with the longer aging time. In addition, phytoplankton growth for cell generation in EDTA-Fe(III) complex media in the presence of insufficient Mn (0 and 5 nM) is strongly influenced by the bioavailable iron supply through the dissociation of EDTA-Fe(III). These results may suggest that T. weissflogii in longer aged am-Fe(III) medium and in EDTA-Fe(III) medium with a higher ratio of EDTA:Fe(III) is in Fe-limitation of growth, which probably increases the production rate of the reactive oxygen species (ROS), and the corresponding up-regulation of the superoxide dismuting enzyme Mn-SOD increases the requirement for Mn.  相似文献   

8.
Pygidiopsis macrostomum and Ascocotyle (Phagicola) pindoramensis (Digenea: Heterophyidae) parasitize guppies as intermediate hosts and, respectively, fish‐eating mammals or birds as definitive hosts. Heterophyids have zoonotic potential, and molecular studies associated with morphological and ecological aspects have helped to clarify their taxonomy and phylogeny. Poecilia vivipara naturally parasitized by metacercariae of both species (100% prevalence) exhibit no external signs of parasitism. In this work, four new sequences of Pmacrostomum (18S rDNA, 28S rDNA and ITS2 rDNA) and one new sequence of A. (P.) pindoramensis (mtDNA cox‐1) are presented. Phylogeny reconstructions linked Pmacrostomum to other heterophyids, but the separation of the Heterophyidae and Opisthorchiidae remains unclear. Additionally, we used indirect immunocytochemistry and the phalloidin‐fluorescence techniques allied with confocal laser scanning microscopy to describe muscular and neuronal structures of Pmacrostomum. A complex arrangement of muscular fibres is associated with the tegument, suckers, gut and reproductive system. Radial fibres around the ventral sucker are thick, branched and extend to the body wall. High‐resolution confocal imaging revealed a typical digenean muscular arrangement and important heterophyid morphological traits. These data will support future control measures to reduce the parasitism in guppies reared in fish farming systems, especially for aquarium and experimental purposes.  相似文献   

9.
为探讨DNA序列标记技术在坛紫菜种质鉴定中的应用,对10个野生坛紫菜种质材料的5.8S rDNA-ITS区进行PCR扩增和序列分析,结果发现扩增的片段长度在1 208~1 219 bp之间,可以分为ITS1区,5.8S区和ITS2区3个部分,其中5.8S区片段的长度完全一致,均为160 bp;ITS1区和ITS2区片段的长度也非常接近,只有几个碱基的差异。多重序列比对发现10个种质材料的ITS区(包括ITS1和ITS2)序列都存在一定差异,序列同源性在95.82%~99.73%之间,而5.8S区序列则完全一致,但与其它种紫菜的5.8S区序列有很大差异,序列同源性在79.7%~95.0%之间。由此认为5.8S rDNA-ITS区这种高度保守区和高变区交替排列的形式可以成为坛紫菜种质鉴定及系统进化分析的强有力工具。  相似文献   

10.
黄姑鱼染色体识别与重复序列定位   总被引:5,自引:4,他引:1  
郑娇  曹款  杨安冉  张静  王志勇  蔡明夷 《水产学报》2016,40(8):1156-1162
黄姑鱼是我国重要的海水经济鱼类。然而,由于细胞遗传标记匮乏,黄姑鱼染色体仍然难以辨识。为了提高黄姑鱼染色体的配对识别水平,本研究利用荧光原位杂交(fluorescence in situ hybridization,FISH)、吉姆萨染色和荧光染色技术分析了黄姑鱼染色体的特征。以总DNA为探针进行基因组DNA荧光原位杂交(genomic fluorescence in situ hybridization,GISH),从而获得黄姑鱼染色体图谱,可使每对染色体呈现特定的荧光信号。依据GISH荧光信号分布模式,可以辨识黄姑鱼的24对染色体。18S r DNA FISH结果显示,18S r DNA只有一对信号,分布于1号染色体臂间,并与吉姆萨染色呈现的次缢痕、DAPI阴性带和DPI染色高亮区域同位。5S r DNA有一强一弱两对信号,信号强的一对分布于1号染色体着丝粒端,信号弱的一对分布于4号染色体的远端。端粒信号在所有染色体的端部显示,但个别染色体一端信号微弱。本研究结果丰富了黄姑鱼的细胞遗传标记,为解决黄姑鱼染色体辨识问题提供参考依据,也为进一步研究石首鱼科染色体进化提供了资料。  相似文献   

11.
Elongate plasmodia with myxosporean spores belonging to the genus Unicapsula, Davis, 1924 were found in the skeletal muscle of the striped seabream, Lithognathus mormyrus (L.), a candidate for the mediterranean aquaculture. The only species of Unicapsula described from the Mediterranean is Unicapsula pflugfelderi Schubert et al. 1975, which occurs in the picarel, Spicara smaris (L.). For morphological and molecular comparison of U. pflugfelderi from S. smaris with Unicapsula sp. from L. mormyrus measurements of plasmodia and spores, ultrastructural details and 18S and 28S rDNA sequences were analysed. Whereas plasmodia were 2–3 times larger in S. smaris than in L. mormyrus (length 2.47–0.81 mm; width 0.22–0.09 mm; P = 0.000), spore morphology showed minor differences and both 18S and 28S rDNA sequences were 100% identical identifying the myxozoan as U. pflugfelderi. Scanning electron microscopy of the spores revealed a different shell valve distribution than the one used for the diagnosis of the genus Unicapsula. This resulted in a review of the genus Unicapsula dividing it into two morphological groups of different spore valve arrangement. TEM revealed the presence of a yet undescribed crystalline structure in the sporoplasm of the spores.  相似文献   

12.
This study identified phytase-producing bacteria that were previously isolated from the gastrointestinal tract of Atlantic cod, Gadus morhua and determined its effect on head kidney leukocytes. Out of the 216 bacterial strains tested, the two phytase producers were identified as Pseudomonas sp. and Psychrobacter sp. based on their 16S rDNA sequence. Crude phytase from these two bacterial strains was produced employing the shake flask method. Even though the total protein of the crude phytase was not significantly different for the two bacteria, the phytase activity of the crude enzyme produced by Pseudomonas sp. (97.1 ± 16.7 U) was significantly higher than that of the enzyme from Psychrobacter sp. (75.9 ± 2.4 U). When cod head kidney leukocytes were incubated with the crude phytase (50 μg ml−1), it resulted in enhanced cell proliferation, higher myeloperoxidase, and acid phosphatase activities. Extracellular responses—respiratory burst activity and hydrogen peroxide production were not enhanced by the crude enzyme. As a consequence, the growth of two pathogenic bacteria Aeromonas salmonicida and Vibrio anguillarum was not suppressed by the supernatants obtained from head kidney leukocytes incubated with the crude bacterial phytase. Thus, the enzyme from phytase-producing intestinal bacteria of Atlantic cod can stimulate intracellular head kidney leukocyte activities but not the production of extracellular substances that are involved in antibacterial response. These have implications on the potential use of bacterial phytase as feed supplement to boost cellular immune response of the fish and could be employed as a health management strategy in culture systems.  相似文献   

13.
以相应引物经PCR扩增了太平洋牡蛎 (Crassostreagigas)的核糖体转录间区域 (ITS 1和ITS 2 )及线粒体 16SrDNA和COI基因片段。PCR产物经T 载体连接后进行克隆和测序 ,分别得到长度为 5 4 3、791、5 30和 70 0bp的核苷酸序列。 4个DNA片段的A、T、G和C碱基含量分别为 2 3.5 7%、2 0 .0 7%、2 9.4 7%和 2 6 .89% (ITS 1) ,2 7.4 3%、19.2 2 %、2 7.0 5 %和2 6 .30 % (ITS 2 ) ,2 9.2 5 %、2 9.2 5 %、2 3.0 2 %和 18.4 9% (16SrDNA) ,2 2 .71%、39.4 3%、2 0 .4 3%和 17.4 3% (COI)。实验证明ITS 1和ITS 2引物在贝类中通用性良好。文中同时讨论了 4个序列在我国几种牡蛎的种类鉴别及相关研究的应用潜力  相似文献   

14.
To construct high-quality 16S rDNA clone libraries for microbial communities associated with Porphyra yezoensis and to minimize the detection of rDNA from leafy gametophytes of P. yezoensis, we designed a new 16S rDNA universal primer (75F). Of the clones prepared using 75F, which was designed to distinguish between bacteria and P. yezoensis, 95% were classified into four groups, namely, β-proteobacteria, γ-proteobacteria, Lentisphaerae, and Flavobacteria. PCR-based analysis of the 16S rDNA primer constructed in this study can be used to implement 16S rDNA-based methodologies for the investigation of microbial community composition and diversity related to the Porphyra group.  相似文献   

15.
核糖体基因为串联重复多拷贝的基因,包括3个编码基因(18S,5.8S,28S)和两个间隔区ITS1(internal transcribed spacer 1)和ITS2(internal transcribed spacer 2)。目前,对核糖体基因的相关报道主要集中在个体内不同拷贝间的多态特征,以及其作为分子标记在系统演化关系中的应用,GC含量作为一项非常重要的核苷酸序列指标,而鲜有报道。为了探讨鱼类的核糖体基因GC含量特征以及间隔区是否也存在GC平衡现象,本研究选择了鲈形目(Perciformes)5科11种鱼类5个片段的核糖体基因进行研究,包括尖吻鲈科(Latidae)、射水鱼科(Toxotidae)、军曹鱼科(Rachycentridae)、剑鱼科(Xiphiidae)、鲹科(Carangidae)。获得了1651个单克隆序列,通过分析并比较已有的其他硬骨鱼序列片段的GC含量变化特征,结果发现:本研究鱼类的18S的GC含量为52.6%~57.1%(平均54.6%),5.8S为55.6%~58.9%(平均57.4%),28S为64.2%~65.8%(平均64.6%),ITS1为56.5%~73.0%(平均65.0%),ITS2为62.3%~77.5%(平均69.1%)。编码区的GC含量相对较保守,变异范围较小,18S和5.8S变化范围明显小于间隔区,28S则位于间隔区的最低值和最高值之间。因此,我们发现硬骨鱼核糖体ITS高于60%的GC含量是该类群的一个特征,并且高GC含量的ITS1和ITS2序列中不存在明显的高GC富集区,其含量高低的变化与序列长度也没有相关性。本研究11种鱼类的ITS1和ITS2的GC含量在种内的相似性既有大于也有小于种间相同片段的相似性,因此GC平衡现象只存在部分种类中。本研究结果可为鱼类核糖体基因序列特征的进一步研究及利用提供科学依据。  相似文献   

16.
The presence of zoonotic Hysterothylacium larvae in fish from Spanish Atlantic and Mediterranean waters, which can cause economic losses for commercial fisheries, has been reported in several studies; however, little is known about species identity in this region. The aim of this study was to identify at species level the Hysterothylacium morphotypes detected in three commonly consumed fish: horse mackerel (Trachurus trachurus), blue whiting (Micromesistius poutassou) and anchovy (Engraulis encrasicolus). Third‐ and fourth‐stage Hysterothylacium larvae, as well as adults obtained from larval in vitro culture, were morphologically and molecularly identified by ITS1/ITS2 rDNA sequencing. Four Hysterothylacium morphotypes were detected. Genetic analysis showed that morphotypes VIII and IX were different larval stages of Hysterothylacium aduncum, which was supported by cultured adult species identification. Morphotypes III and IV were found to correspond to different developmental stages of another species of Hysterothylacium. As all larval types detected were morphologically indistinguishable from others previously reported yet showed clear genetic differences, they are referred here as new genotypes. This is the first time that ITS‐sequence data of various developmental stages of the same species, including adults, have been studied and compared, providing crucial knowledge for future studies on Hysterothylacium identification and biology.  相似文献   

17.
利用荧光原位杂交(FISH)技术将5S rDNA基因定位在牙鲆和半滑舌鳎染色体上,结果表明,5S rDNA基因在雌、雄性半滑舌鳎的一对同源染色体上分别存在2个杂交信号位点;在二、三倍体牙鲆的同源染色体上分别存在2个和3个杂交信号位点,杂交信号明显且特异。本研究首次将5S rDNA基因定位在半滑舌鳎和三倍体牙鲆的染色体上,为牙鲆及半滑舌鳎倍性鉴定、染色体鉴别提供了有效方法。此外,根据牙鲆5S rDNA基因编码区序列设计引物,扩增出大菱鲆和半滑舌鳎5S rDNA基因编码区序列,与鲽、塞内加尔鳎、大口黑鲈、七鳃鳗的5SrDNA基因序列进行同源性比较后发现,5种鲽形目鱼类5S rDNA基因序列同源性高达98.3%,系统发生分析结果显示5种鲽形目鱼类聚为一支;各序列中GC含量均显著高于AT含量。  相似文献   

18.
19.
To identify the pathogens causing saprolegniosis among farmed fish in Nova Scotia, 172 infected tissues and 23 water samples were collected from six species of teleosts: Atlantic salmon (Salmo salar), brown trout (Salmo trutta), Arctic charr (Salvelinus alpinus), brook trout (Salvelinus fontinalis), striped bass (Morone saxatilis) and rainbow trout (Oncorhynchus mykiss) at nine facilities over a 600 km range. Following laboratory culture, 132 isolates were recovered. Six species of oomycetes were identified from analysis of the internal transcribed spacer (ITS) sequence of the nrDNA: Saprolegnia parasitica, Saprolegnia ferax, Saprolegnia diclina, Saprolegnia aenigmatica, Saprolegnia torulosa, Saprolegnia sp. and Pythiopsis cymosa. Further phylogenetic analyses of the ITS and cytochrome c oxidase subunit 1 (Cox1) regions revealed four strains of Saprolegnia parasitica (named here as S1, S2, S3 and S4), of which S1 and S2 were common (37% and 42% of the isolates), and two strains of S. ferax. Among S. parasitica, S2 and S3 are more closely related to each other than to S1 based on the phylogenetic analyses and predicted RNA secondary structure of the ITS region. Sexual structures with a similar morphology were formed by S1 and S3 in vitro, but were not formed by S2.  相似文献   

20.
A disease outbreak occurred in swimming crab (Portunus trituberculatus) farmed in eastern China, with a mortality rate of more than 80%. To further investigate the characteristics and pathogenesis, we reported isolation, characterization and virulence of the causative agent of this disease from 10 sick crabs. Histopathological observation found that multiple tissues, especially haemolymph, contained lots of ciliates. The ciliate was isolated and cultured in vitro, and molecular and morphological studies were done. The results showed that SSU rDNA and LSU rDNA sequences of the ciliate were similar to Mesanophrys ciliates (>96.81%), while ITS1-5.8s-ITS2 sequence was similar to Mesanophrys pugettensis (95.37%) and identical to Orchitophrya stellarum (100%). Furthermore, the results of the morphological study confirmed that the ciliate was similar to Mesanophrys ciliates and O. stellarum cultured in supportive media, but different from O. stellarum cultured in living sperm cells of starfish (Leptasterias spp.). Also, the growth of the ciliate did not interfere with light, which was different from O. stellarum. Accordingly, the ciliate was classified as genus Mesanophrys and temporarily named as Mesanophrys sp. In addition, experimental infection confirmed that Mesanophrys sp. was the pathogen that infected farmed crabs. In summary, Mesanophrys sp. was first isolated and characterized in P. trituberculatus.  相似文献   

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