Molecular cloning,sequence characterization,and expression analysis of C-type lectin (CTL) and ER-Golgi intermediate compartment 53-kDa protein (ERGIC-53) homologs from the freshwater prawn,Macrobrachium rosenbergii

Aquaculture International - Lectin protein families are diverse and multi-functional in crustaceans. The carbohydrate-binding domains (CRDs) of lectins recognize the molecular patterns associated...     

Localization of carbohydrate terminals on Tetracapsuloides bryosalmonae using lectin histochemistry and immunogold electron microscopy

Tetracapsuloides bryosalmonae is the myxozoan parasite that causes the commercially important proliferative kidney disease (PKD) in salmonid aquaculture. Previous studies on the binding of lectins to T. bryosalmonae identified Griffonia simplificola agglutinin I (GS I) as useful for parasite identification. This lectin was also implicated as recognizing antigenic structures on the parasite. Here, we examine the histochemical staining and ultrastructural localization of a panel of 21 lectins on the extrasporogonic stage of T. bryosalmonae. The histochemical staining studies indicated that the majority of lectins bound to the renal stages of T. bryosalmonae, however not all of these lectins could be successfully localized using immunogold electron microscopy. Of the lectins that were localized many, including GS I, bound to membranes associated with the lysosomal pathway within the extrasporogonic primary cell of the parasite, indicating that these organelles are rich in glycoconjugates. The histochemical staining of Erythrina cristagalli ECL was unique and highlighted a different distribution of glycoconjugates in the periphery of some extrasporogonic parasites within the renal sinuses when compared with stages in the interstitium, suggesting the presence of distinct blood forms of T. bryosalmonae.     

Different expression of Litopenaeus vannamei (Boone) haemocytes to Vibrio and abiotic particle inoculation

Penaeid shrimps are among the most studied crustaceans, mainly regarding their immune system, and several proteins involved in the defense against pathogens have been described. Haemocytes are very dynamic cells responsible of recognition, phagocytosis, degranulation and nodule formation, but still little is known about their gene expression. Using differential display, we found modification in haemocyte gene expression after inoculation with an abiotic particle (DEAE‐Sephadex) or potential pathogenic bacteria (Vibrio alginolyticus). We also noticed that some of the newly expressed genes are exclusive from a specific treatment. Here we report that haemocytes from Litopenaeus vannamei are capable of recognizing and distinguishing different foreign materials, and respond specifically to each treatment, indicating some specificity in shrimp immune response.     

Characterization of glycans in the developmental stages of Myxobolus cerebralis (Myxozoa), the causative agent of whirling disease

Glycans and sugar-binding molecules (lectins) form an interactive recognition system, which may enable parasitic organisms to adhere to host cells and migrate into target tissues. The aim of the present study was to analyse surface-associated glycans in the developmental stages of Myxobolus cerebralis (Hofer), the causative agent of whirling disease. A panel of biotin-labelled plant lectins was used to detect a broad spectrum of glycan motifs with high specificity. Binding sites were detected histochemically in the tissue sections of infected rainbow trout, Oncorhynchus mykiss (Walbaum), and infected Tubifex tubifex (Müller), and were characterized by light, fluorescence and transmission electron microscopy. With mannose-specific lectins [Lens culinaris agglutinin, Pisum sativum agglutinin, Canavalia ensiformis agglutinin (LCA, PSA, CanA)] mannose-containing glycans were detected in all the developmental stages and host tissues. No binding sites for galactose-specific lectins were present in M. cerebralis spores but reactivity with host tissues occurred. Diversity in glycans was detected by N-acetyl-D-galactosamine-specific lectins in sporoplasm cells of M. cerebralis and triactinomyxon spores. In the group of lectins with monosaccharide-specificity for N-acetyl-D-glucosamine (GlcNAc), the reactivity of Datura stramonium agglutinin (DSA), Lycopersicon esculentum agglutinin (LEA) and Solanum tuberosum agglutinin (STA) was restricted to polar capsules whereas Griffonia simplicifolia agglutinin II (GSA II) also bound to sporoplasm cells of stages in the fish host but not in those present in infected T. tubifex. Moreover, Triticum vulgaris (wheat germ) agglutinin (WGA) and succinylated WGA indicated the presence of N-acetyl-D-glucosamine polymers in polar capsules. No specificity for spores was observed concerning 'bisected'N-glycans and no reactivity in parasitic stages was observed with the fucose-binding lectin Ulex europaeus agglutinin (UEA) I, Sambucus nigra agglutinin (SNA) (specific for alpha2,6-sialylated glycans) and Maackia amurensis agglutinin (MAAI) (specific for alpha2,3-sialylated glycans). Arachis hypogaea (peanut) agglutinin (PNA), Erythrina cristagalli agglutinin (ECA), GSA I, Sophora japonica agglutinin (SJA), Dolichos biflorus agglutinin (DBA) and GSA II detected reactive sites solely confined to the developmental stages of M. cerebralis and were not reactive in the fish host. These parasite-specific glycans may play a role in the adhesion process of the parasite to fish epidermis prior to infection, but may provide protection to the host by activating the complement system, or stimulating an adaptive immune response as putative antigens.     

Bioactive roles of carotenoids and retinoids in crustaceans

Among the proposed functions of carotenoids in aquaculture have been those of pigmentation, antioxidant functions, as a source of pro-vitamin A, cellular protection from photodynamic damage, enhancement of growth and reproductive potential. However results reported by various authors are often contradictory. Some evidence suggests that these pigments may perform vital roles in growth and reproductive success in Crustaceans. Additional efforts need to be devoted to the understanding of the active forms of the carotenoid derived metabolites, retinoids in crustaceans. Dietary carotenoids are the sole biological precursors of retinoids in crustaceans. The importance of carotenoids as bioactive molecules reside to a large degree on their conversion to retinoids that are involved in the activation of hormonal nuclear receptors. Retinoids play a prominent role in many developmental processes, including embryonic development and differentiation of various cell types. The presence of receptors of retinoic acid in crustaceans and our findings of retinoids in the neuroendocrine complex and in reproductive tissue, as well as the enhancement of the ovarian development in shrimp suggests an important role of these metabolites in shrimp physiology for their successful aquaculture.     

Cell membrane glycoconjugates on virulent and avirulent strains of the haemoflagellate Cryptobia salmositica (Kinetoplastida)

The glycoconjugates on the cell membranes of Cryptobia salmositica (virulent and avirulent strains) were analysed using 13 highly purified lectins. The virulent strain of C. salmositica was agglutinated by two of these lectins (Concanavalin A (Con A) Canavalia ensiformis , specific for α-mannose and α- D -glucosyl residues; Pisum sativau agglutinin (PSA), specific for N-acetyl α- D -galactosaminyl) but the avirulent strain of C. salmositica was agglutinated by 11 lectins. No agglutination of C. salmositica (virulent and avirulent strains) was observed with lectin Tetragonolobus purpureas agglutinin (TPA, specific for α- L -fucose). Glycoprotein analysis with digoxigenin or biotin labelled lectins showed strain specific staining patterns for C. salmositica; with the avirulent strain showing stronger reactions than the virulent strain. These results indicate that the surface carbohydrate residues changed with attenuation of the pathogen and this change may be related to its loss of virulence.     

泛甲壳动物激素调控卵黄蛋白原合成的分子机制研究进展

昆虫和甲壳动物由于进化上亲缘关系近统称泛甲壳动物，大多数泛甲壳动物卵黄蛋白原为雌性特异性蛋白，是卵母细胞中储存的卵黄蛋白的前体，可为胚胎发育提供营养和能量，是决定繁殖性能的关键因素，在生殖发育起着十分重要作用。业已证明，调控泛甲壳动物卵黄蛋白原的主要激素有保幼激素、蜕皮激素、神经肽和胰岛素样肽等。昆虫保幼激素和蜕皮激素发挥促进作用，而神经肽和胰岛素样肽对卵黄蛋白原合成的调控作用因种类不同存在不同。除了以上激素，甲壳动物特有的眼柄高血糖激素家族以及促雄腺激素对卵黄蛋白原的合成起负调控作用。本文总结了近年来泛甲壳动物卵黄蛋白原合成的调控机制的相关研究，概括并比较了调控昆虫与甲壳动物的卵黄蛋白原合成的主要激素及调控机制，为养殖虾蟹甲壳动物生殖调控相关研究提供参考。     

Carbohydrate and lectin interactions with Edwardsiella ictaluri and channel catfish, Ictalurus punctatus (Rafinesque), anterior kidney leucocytes and hepatocytes

Abstract. Seven isolates of Edwardsiella ictaluri were evaluated for channel catfish red blood cell and neutrophil agglutination properties and reactivity with lectins and carbohydrates. All isolates were agglutinated by lectin derived from Ricinus communis and the intensity of the agglutination reaction could be lessened or abrogated by premixing the lectin derived from Ricinus communis with β-D-galactose or α-L-fructose. Edwardsiella ictaluri isolates also sedimented much more rapidly when mixed with lectin derived from Ricinus communis than when mixed with other lectins. Premixing Ricinus communis with β-D-galactose had the greatest effect on slowing the rate of sedimentation. Two out of the seven E. ictaluri isolates were capable of agglutinating channel catfish red blood cells and anterior kidney neutrophils, and this agglutination could be prevented by the addition of β-D-mannose. Likewise, the per cent phagocytosis of E. ictaluri isolate 1389 (the most intense neutrophil agglutinator) could be dramatically reduced by the inclusion of mannose in the assay mixture. Anterior kidney leucocytes and hepatocytes primarily reacted with lectins derived from Canavalia ensiformis, Ricinus communis and Triticum vulgaris. The presence on channel catfish cells of carbohydrate-lectin receptors similar in function to those found on mammalian cells and the reaction of E. ictaluri with specific lectins suggest that bacterial clearance and lectinophagocyotosis may occur in a similar manner in catfish to that in mammals.     

Advances in precocity research of the Chinese mitten crab <Emphasis Type="Italic">Eriocheir sinensis</Emphasis>

Precocity has been observed in fishes and crustaceans. However, mechanisms underlying precocity have not been well documented in crustaceans and are thought to be influenced by both genetic and environmental factors. Also, precocity is generally considered to have negative effects on crustaceans. The Chinese mitten crab Eriocheir sinensis, a catadromous species endemic to China, is a high valued commodity and in the recent past is being extensively cultured to meet the growing demand by the restaurant trade. The mitten crab is an ideal candidate for precocity studies because of their large size, distinct secondary sex characters, wide distribution and abundant availability from commercial farms. In this article, progress in several aspects of precocity of E. sinensis is reviewed, including the phenomena of precocity and its effect, identification of precocious crabs, factors related to precocity (temperature, salinity, light, nutrition, stocking density, and germplasm), relationships between precocity and neuro-endocrine system, steroid hormones or hepatopancreas, prevention and control methods of precocity. In addition, possible future directions for the study of precocity are suggested.     

唐山湾海域渔业资源群落多样性分析

基于2012年5、8和10月对唐山湾海域进行底拖网渔业资源调查的资料,采用相对重要性指数、多样性指数及群落ABC曲线等分析方法,对3个季节渔业资源现状及群落多样性进行研究。结果显示:渔获物中共有海洋动物59种,其中鱼类27种,甲壳类17种,贝类7种,棘皮动物4种,头足类3种,多毛类1种;春季捕获35种渔获物,夏季36种,秋季37种;春季平均渔获率(质量)为3.50 kg/h,夏季为13.68 kg/h,秋季为16.99 kg/h。研究表明:(1)渔获率及其组成季节差异明显。在生物量上,春季以头足类为主,鱼类与甲壳类次之;夏季甲壳类为主,鱼类次之;秋季鱼类为主,甲壳类次之。(2)渔获率空间分布季节变化明显,春季浅水区较深水区渔获率高,夏季深水区较浅水区高。(3)优势种季节变化不明显。(4)春季H'高于夏、秋季,深水区总体高于近岸浅水区。(5)ABC曲线分析可知,群落受到严重干扰。     

中国对虾不同凝集素基因应答WSSV侵染的表达差异

本研究探讨了7种细胞凝集素基因在中国对虾中应答白斑综合征病毒(WSSV)感染上的生物学功能差异.结果显示,LT1 (DQ167572.1)、LT2(EU834289.1)、LT3 (EU834292.1)和LT5(EU834290.1)是肝胰腺组织特异、受WSSV感染诱导表达的基因,其他3种基因主要在肌肉、肠和腮中表达,与WSSV感染诱导无明显相关;在应答WSSV感染不同时间段的表达特征上,7种凝集素基因各不相同;在各组织中的表达量上,方差分析表明7种凝集素基因除在肝胰腺中的表达上存在显著差异外,其他组织中不存在显著差异.这些结果显示这7个凝集素基因在应答WSSV侵染的表达特征上存在较大差异,推测其在中国对虾体内免疫防御功能上的作用也各不相同.     

Cryopreservation of aquatic invertebrate semen: a review

Research on cold storage and cryopreservation of aquatic invertebrate spermatozoa is reviewed in this paper. A few studies are available on the cryopreservation of aquatic invertebrate semen. These have been limited to echinoderms (sea urchin, sand dollar and starfish), molluscs (oyster and abalone), polychaetes and crustacea (shrimp and crab). In fact, only about 30 species have been investigated to date, and research has been primarily concentrated on the spermatozoa of the oyster. Although aquaculture of decapod crustaceans has become very important, information on sperm biology is limited, and sperm preservation techniques are sparse. The sperm of decapod (shrimp and crab) crustaceans are aflagellate and non‐motile, and eggs are not readily available. One of the main reasons for the problems in the cryopreservation of crustacean sperm is considered to be the lack of reliable viability evaluation. Conclusions and new avenues of future research are proposed.     

SNP分子标记技术在经济甲壳动物中的应用进展

作为第三代分子标记,单核苷酸多态性标记(SNP)具有丰富度高、密度大、稳定性强、共显性等特点,成为目前虾、蟹等经济甲壳动物中应用最广泛的分子标记技术。本文对经济甲壳动物中SNP技术的发展及其在高密度遗传连锁图谱的构建、种质资源遗传多样性分析、分子辅助育种等方面的应用进展进行了综述,并提出了未来有待加强的研究方向,以期为经济甲壳动物种质资源的开发和利用提供理论指导。     

Histochemical and immunohistochemical characterization of rodlet cells in the intestine of two teleosts,Anguilla anguilla and Cyprinus carpio

Rodlet cells (RC) are characterized by a distinctive cell cortex and conspicuous inclusions named “rodlets.” These cells are particularly abundant and large in size in intestine of eels. Histochemical, immunohistochemical and ultrastructural investigations were carried out on European eel Anguilla anguilla and Common carp Cyprinus carpio from Northern Italy. Eight biotinylated lectins were used to probe for specific carbohydrate residues in deparaffinized, hydrated intestinal sections of eel and carp. Five antibodies were tested on intestinal sections of both fish species: inducible nitric oxide synthase (i‐NOS), leu‐enkephalin, lysozyme, serotonin and tumour necrosis factor‐α. Lectin histochemistry revealed rodlet cells (RCs) of the eel intestine to react with two of the eight lectins tested, specifically Concanavalin A (ConA) and Sambucus Nigra Agglutinin (SNA). This contrasted to lectin staining of RCs in the intestine of common carp, where four of the eight lectins showed a positive reaction; Dolichos Biflorus Agglutinin (DBA), Wheat Germ Agglutinin (WGA), SNA and ConA. RCs in eel and carp intestine were immunoreactive with antibodies to lysozyme and i‐NOS. The occurrence of the inflammatory peptides lysozyme and i‐NOS in RCs of the eel and common carp poses in favour that these cells are involved in the mechanism of defence against pathogens.     

厦门附近几种浮游甲壳动物的化学成分

1.本文分析的几种海洋浮游甲壳动物都含有占干物质50％以上的蛋白质,氨基酸的组分也相当完整。其中必需氨基酸的含量相当丰富,特别是色氨酸。如桡足类的色氨酸含量比牛肉多了三分之二。 2.经过加工处理的甲壳动物干制品,无论是蛋白质或必需氨基酸的含量损失不大,基本上保持其原有的营养值。 3.一般浮游甲壳类比底栖型的甲壳类,所含的脂肪多而灰分的含量正相反,这是营浮游生活的一种适应现象。 实验结果表明,以上几种海洋浮游甲壳动物,无论作为人类的食品或其他海洋经济动物的饵料,都具有很高的营养价值,在渔业上具有重大的意义。     

黄渤海春季甲壳类群落结构的空间变化

为了解黄渤海甲壳类动物资源状况, 于2010年5月采用底拖网调查, 对黄渤海甲壳类动物的群落结构进行了研究。结果显示,黄渤海共捕获甲壳类32种, 其中虾类18种, 蟹类13种, 口足类1种; 不同海区优势种类组成差异较大, 脊腹褐虾在各海区均为优势种, 是黄渤海生态系统中最重要的甲壳类物种之一; 对黄渤海各海区甲壳类平均相对资源密度组成进行研究, 各海区甲壳类生物量均以蟹类密度最高, 黄渤海蟹类平均相对资源量为2.97 kg/h, 丰度均以虾类密度最高, 黄渤海虾类平均资源密度为1 825 ind/h; 对黄渤海甲壳类生物量的空间分布进行研究, 各海区生物量排序为黄海北部>黄海南部>黄海中部>渤海; 对黄渤海各海区甲壳类的生物多样性进行研究, 无论根据生物量还是丰度, 黄海南部的多样性指数最高, 黄海中部的多样性指数最低; 对各海区甲壳类群落结构相似性进行研究, 毗邻海区甲壳类群落结构的相似性指数相对较高。     

眼柄摘除对红螯光壳螯虾生长、性腺发育及体色的影响

为探究眼柄摘除对红螯光壳螯虾(Cherax quadriarinatus)生长、性腺发育及体色的影响,对6月龄红螯光壳螯虾[体质量(47.47±3.48)g]进行了摘除眼柄处理实验。结果显示,4周后摘除双侧眼柄组红螯光壳螯虾体质量为(84.40±13.41)g,显著高于不摘除眼柄组[(49.63±6.47)g]和摘除单侧眼柄组[(51.47±4.08)g](P<0.05);摘除双侧眼柄组红螯光壳螯虾存活率为60.0%±4.8%,显著低于不摘除眼柄组和摘除单侧眼柄组(分别为94.4%±2.3%和97.2%±2.1%);摘除双侧眼柄组红螯光壳螯虾性腺指数为1.23±0.50,显著高于不摘除眼柄组(0.20±0.06)和摘除单侧眼柄组(0.35±0.08)。利用ImageJ软件对红螯光壳螯虾体色RGB值分析表明,摘除双眼柄组红螯光壳螯虾体色G值和B值(分别为99.26±5.23和98.40±3.58)显著高于摘除单侧眼柄组(59.02±3.85和44.07±4.57)(P<0.05)。结果表明,眼柄摘除能促进红螯光壳螯虾生长和性腺发育,影响体色变化。     

Rhamnose-binding lectins induce respiratory burst activity in macrophage cells from rainbow trout

Rhamnose-binding lectins (RBLs) have been isolated and characterized from various kinds of fish. RBLs recognize the structures of lipopolysaccharides and lipoteichoic acid on microbes, and have an opsonic effect, suggesting that RBLs play an important role as pattern recognition proteins in innate immunity. However, the function of RBLs in the removal of pathogens from the body has not yet been elucidated. In this study, we investigated the effect of three RBLs—named CSL1, 2, and 3—that were isolated from chum salmon Oncorhynchus keta eggs on the respiratory burst activity in a macrophage cell line. The CSLs induced reactive oxygen species in the peritoneal macrophage cell line from rainbow trout Oncorhynchus mykiss (RTM5). However, this activity was not affected by l-rhamnose or dl-threo-1-phenyl-2-palmitoylamino-3-morpholino-1-propanol, an inhibitor for glucosylceramide synthesis, indicating that the interaction between CSLs and Gb3 on cell surfaces was not involved in the induction of respiratory burst activity.     

The effects of holding space on juvenile red king crab,Paralithodes camtschaticus (Tilesius, 1815), growth and survival

Rearing crustaceans communally for aquaculture, stock enhancement or research often results in high rates of cannibalism and low yields. One potential strategy to reduce loss from cannibalism is to rear crustaceans in individual cells. As small holding cell size can result in decreased growth or increased mortality, it is essential to identify the optimal holding cell size, both for mass culturing efforts and for experimental design purposes. In this study, we reared juvenile red king crab, Paralithodes camtschaticus (3.67–8.30 mm carapace length) in 20, 40 and 77 mm diameter holding cells, and monitored growth and survival over a 274‐day experiment. A trend of lower growth per molt in the smallest holding cells resulted in crab 17% smaller than those in the large holding cells at the end of the experiment. In addition, mortality rates were an order of magnitude higher in the small holding cells compared with the large or medium cells. For individual rearing of this size of juvenile red king crab, the medium‐sized cells (40 mm diameter) are the optimal size as there was no increase in mortality and only marginally lower growth rates compared to the large‐sized cells.