An Invasive Shrimp Species,Machrobrachium nipponense,in Anzali Wetland Demonstrated a Potential Source for Commercial Fishing

ABSTRACT

The proximate content, fatty acids composition, and nutritional quality index (NQI) of Macrobrachium nipponense at three habitats in the Anzali wetland in Iran were investigated as a potential source for human consumption. The highest amounts of protein, lipid, ash, and energy contents in muscle of M. nipponense were showed in autumn (non-reproductive season) (p < 0.05). The main monounsaturated fatty acids (MUFA) were oleic acid (C18:1 n9 C, C18:1 ω9 T) and palmitoleic acid (C16:1). Moreover, the main polyunsaturated fatty acids (PUFA) were docosahexaenoic acid (DHA, C22:6 n3), eicosapentaenoic acid (EPA, C20:5n3), arachidonic acid (ARA, C20:4 n6), linoleic acid (LA, C18:2 ω6), and α-linolenic acid (ALA, C18:3 n3). The predominant individual saturated fatty acid (SFA) was palmitic acid (0.07–13.4%), while oleic acid (14.7–26.3%), EPA (3.5–12.7%) and linoleic acid (0.04–14.9%) represented the most abundant individual MUFA and PUFA in M. nipponense. The highest mean value of EPA+DHA (14.0), n3/n6 (1.02), ΣMUFA/ΣSFA (1.05), ΣPUFA/ΣSFA (1.04), and EPA/DHA (3.8) ratios in M. nipponense was in autumn. The range of atherogenicity index (AI) and thrombogenicity index (TI) was much lower, from 0.42 to 0.6 and from 0.33 to 0.57, respectively, in terms of season. The results obtained in the present study show that M. nipponense is an excellent nutritional food source in the Anzali wetland.     

The androgenic gland and monosex culture of freshwater prawn Macrobrachium rosenbergii (De Man): a biotechnological perspective

Males of the freshwater prawn Macrobrachium rosenbergii (De Man) grow faster and reach a larger size at harvest than females of the species. It is thus obvious that culture of monosex all‐male populations would be economically advantageous. Sexual differentiation in crustaceans is regulated by the androgenic gland (AG), which plays a pivotal role in the regulation of male differentiation and in the inhibition of female differentiation. In M. rosenbergii, AG removal from immature males resulted in sex reversal, with complete female differentiation. Similarly, AG implantations into immature females lead to the development of the male reproductive system. Sex‐reversed M. rosenbergii animals were capable of mating with normal specimens to produce offspring. Early attempts in Israel and more recently, attempts in other countries to establish all‐male populations through manual segregation showed that for the production of monosex prawn populations to be economically feasible, intervention via the AG is probably required. However, a suitable biotechnology is still to be developed, and an androgenic hormone has yet to be identified in decapods. Three lines of aquacultural and biotechnological research and development are proposed for the future: (1) Establishment of monosex cultures through manual segregation, together with the application of selective harvesting and claw ablation, as well as examination of different monosex culture strategies under a variety of economic conditions. (2) Microsurgical intervention in the AG, leading to the development of functional neo‐females, which would subsequently be mated with normal males to produce all‐male progeny. (3) Elucidation of AG bioactive products to enable biochemical or molecular manipulation of sex differentiation.     

慢性盐度胁迫下金钱鱼卵巢转录组分析

为了阐明盐度胁迫对金钱鱼(Scatophagus argus)代谢和生殖的影响,采用RNA-seq技术对低盐组(5)、对照组(25)和高盐组(35)处理40 d后的2龄性成熟金钱鱼卵巢进行转录组分析。结果发现,金钱鱼卵巢转录组测序获得raw reads共398681318,clean reads共396910398。从低盐胁迫相对对照组(5 vs 25)和高盐胁迫相对对照组(35 vs 25)中分别筛选到373个和874个差异表达基因(DEGs)。与对照组相比,低盐胁迫后氨基酸代谢相关基因(sds、bhmt)和脂肪酸代谢相关基因(pnpla2)显著下调;高盐胁迫后pgr、cyp17a1和ers1等生殖相关基因显著下调。KEGG通路富集分析发现,低盐胁迫组相对对照组显著富集与代谢相关的通路为半胱氨酸和甲硫氨酸代谢,缬氨酸、亮氨酸和异亮氨酸生物合成代谢,脂肪酸生物合成,脂肪酸代谢,皮质醇的合成和分泌,醛固酮的合成和分泌,脂肪细胞脂解的调节等;高盐胁迫组相对对照组中显著富集的与生殖相关的通路为雌激素信号传导通路。这些结果表明,氨基酸和脂肪酸的代谢调控可能在金钱鱼卵巢的低渗透压调节中起重要作用...     

日本沼虾血清淀粉样蛋白A(SAA)基因的克隆及其免疫功能

为探索血清淀粉样蛋白A(SAA)在日本沼虾免疫防御系统中的作用,实验利用RACE方法克隆了日本沼虾SAA(MnSAA)基因cDNA全长序列,采用实时荧光定量PCR(QPCR)分析其在日本沼虾不同组织、不同发育时期的表达情况,以及溶壁微球菌、嗜水气单胞菌和白斑综合征病毒(WSSV)感染后MnSAA在血淋巴细胞和肝胰腺中的表达情况,同时还利用RNA干扰(RNAi)技术检测了MnSAA基因沉默后,日本沼虾再感染嗜水气单胞菌后其肝胰腺中激活子蛋白-1(AP-1)和B类清道夫受体(CD36)基因表达变化及虾的累积死亡率变化。结果显示,M n S A A基因c D N A全长649 b p(G e n B a n k登录号:MK292888),包含21 bp 5′非编码区,232 bp 3′非编码区,369 bp开放读码框,编码131个氨基酸残基;氨基酸序列比对及系统进化分析结果显示,MnSAA蛋白属于急性期血清淀粉样蛋白A蛋白(A-SAA),在进化上与无脊椎动物香港牡蛎的亲缘关系最近;mRNA表达分析显示,MnSAA基因在日本沼虾的各组织和各生长阶段均有表达,分别在肝胰腺和成虾阶段中的表达量最高;病原体感染实验表明,在日本沼虾感染溶壁微球菌、嗜水气单胞菌和白斑综合征病毒后12~72 h,其血淋巴和肝胰腺中的MnSAA基因表达量与对照组相比均显著升高;RNAi实验显示,MnSAA基因沉默后,日本沼虾肝胰腺中AP-1和CD36基因分别在其感染嗜水气单胞菌后12~72 h和6~72 h与对照组相比显著下降,日本沼虾累积死亡率在感染嗜水气单胞菌后与对照组相比明显升高。研究表明,MnSAA参与日本沼虾的抗病原体感染的反应,在日本沼虾的免疫防御体系中具有重要的作用。     

Effects of androgenic gland ablation on growth,sexual characters and spermatogenesis of the white shrimp,Litopenaeus vannamei (Decapoda: Penaeidae) males

This research was implemented to study the effects of androgenic gland ablation in the white shrimp Litopenaeus vannamei to explore sex reversal potential as an alternative technology for monosex female mariculture based on the sex dimorphic growth of this species. The surgical procedure was applied to male postlarvae (PL) at different ages, after external sex differentiation, as well as in sex undifferentiated PL. Andrectomized males regenerated normal appendices including pereiopods and pleopods; however, body growth and relative size of regenerated petasmas and appendices masculinae were statistically inferior (P ? 0.05) to control males. Spermatogenesis in andrectomized males was active, but a phenomenon of degradation of spermatids and reproductive tissues was detected. No sex reversal was accomplished regardless of PL age from sex undifferentiated stages (PL38) to sex differentiated stages (?PL55). The complete regeneration of sexual characters in andrectomized L. vannamei (Dendrobranchiata) is different from previous reports from Decapoda.     

Molecular characterization of transformer‐2 gene in the mud crab Scylla paramamosain and its putative role in sexual development

Transformer‐2 (Tra‐2) is an mRNA splicing factor that acts in concert with Tra to regulate the female sexual development in Drosophila melanogaster. In decapods, as no Tra homologues have been identified, the role of Tra‐2 in sexual development is more often discussed accordingly. In the present study, the full‐length cDNA of Tra‐2 of the mud crab Scylla paramamosain (SpTra‐2) was cloned and characterized. The deduced amino acid sequence of SpTra‐2 contained an RNA‐recognition motif (RRM) flanking by two RS‐rich regions, which are the hallmarks of the Tra‐2 proteins. The SpTra‐2 mRNA was highly abundant in the hepatopancreas, muscles and androgenic glands of the male crabs, implying a potential role in male sexual development. The hypothesis was supported by the RNAi experiments, finding that the injection of SpTra‐2 dsRNA in vivo led to a decreased expression of SpIAG, a key gene in crustacean male sexual differentiation. The relationship between SpTra‐2 and SpIAG was further affirmed by eyestalk ablation (ESA) experiments, which caused SpTra‐2 expression earlier than SpIAG expression in both androgenic glands and hepatopancreas. In addition, treatment with SpTra‐2 dsRNA also reduced the mRNA level of SpSxl, but had no effects on SpDmrt expression, suggesting the sex determination system in S. paramamosain is different from that in D. melanogaster.     

Effects of salinity acclimation on the growth performance,osmoregulation and energy metabolism of the oriental river prawn,Macrobrachium nipponense (De Haan)

In this study, we evaluated the growth, osmoregulation and energy metabolism of the oriental river prawn, Macrobrachium nipponense, reared during 6 weeks with different salinities (0, 8, 14 and 22 g/L). The results showed that the haemolymph osmolality of M. nipponense increased with an increase in ambient osmotic pressure; the isosmotic point was 490 mOs/kg H₂O. The prawns showed a higher survival rate, weight gain rate and hepatopancreas index in salinity 14 g/L. Digestive enzymes were all affected by salinity, and the highest activities were observed in the salinity 14 g/L. The mRNA expression of Na⁺‐K⁺‐ATPase in gills and p53 in hepatopancreas were the highest in salinity 22 g/L. The expressions of heat shock protein 90 and glutathione S‐transferase genes in hepatopancreas were significantly higher in the salinity 8 g/L. Lipid metabolism‐related genes in hepatopancreas were significantly expressed in the salinity 14 g/L. The glucose‐6‐phosphatase gene in hepatopancreas was highly expressed in the salinity 8 and 22 g/L, and the expression of the ecdysone receptor gene in hepatopancreas was significantly higher in the salinity 14 g/L. The results showed that salinity 14 g/L could promote the growth of M. nipponense. However, higher salinity conditions may cause physiological damage, which provides a theoretical basis for brackish water culture of M. nipponense.     

Molecular cloning,characterization and expression analysis of the fatty acid‐binding protein (MnFABP), involved in dietary lipid sources response in oriental river prawn,Macrobrachium nipponense

A fatty acid‐binding protein (FABP) gene designated as MnFABP10 was cloned and characterized from the freshwater prawn Macrobrachium nipponense. The full‐length cDNA of MnFABP10 was 646 bp encoding a 130 amino acid. Real‐time quantitative RT‐PCR showed that the MnFABP10 gene was expressed in various tissues with the highest expression in the hepatopancreas. The MnFABP10 mRNA levels in the hepatopancreas and ovary of M. nipponense were dependent on the stages of ovarian development. Western blot results revealed a single immunoreactive band with an estimated molecular mass of approximate 14 kDa in the developmental ovary. Then, M. nipponense with an initial body weight of 0.090 ± 0.0010 g were fed with four isonitrogenous and isocaloric diets with different oils, that is, beef tallow (BT), soybean oil (SO), pollack fish oil (FO) and a mixture of fish oil and soybean oil (FO/SO 2 : 1 w/w) for 52 days. The mRNA levels of MnFABP10 in the hepatopancreas were influenced by different lipid sources, with a peak expression observed in prawns fed SO. This study suggests that MnFABP10 may have a putative function in ovary maturation, and its mRNA expression in the hepatopancreas can be regulated by the source of dietary lipids in M. nipponense.     

转录组测序筛选克氏原螯虾卵巢发育、免疫和生长相关基因

为发掘克氏原螯虾卵巢发育、免疫和肌肉生长的重要功能基因,采用Illumina HiSeqTM 2 500高通量测序平台对克氏原螯虾的卵巢、肝胰腺和肌肉组织进行了转录组测序.所得序列经质控、组装后比对到NR、Swiss-Prot、pfam、COG、GO和KEGG数据库中注释,并进行差异基因聚类分析.结果 显示,测序共获得...     

KK-42对蜕皮后期日本沼虾头胸甲超微结构的影响

为了深入探究KK-42对表皮结构的影响,本实验以外表皮形成趋于结束而内表皮开始出现的蜕皮后期幼虾为材料,通过扫描电镜观察石蜡切片法,分析了KK-42处理前后头胸甲外表皮,尤其是内表皮结构的变化。将处于蜕皮间期的健康幼虾[体长(3.5±0.1)cm]随机分为2组,分别用1.95×10^-4 mol/L的KK-42溶液和不含KK-42的溶液处理,取蜕皮后1.5、3.0、6.0及12.0 h幼虾头胸甲,观察其超微结构。另取蜕皮后6.0 h头胸甲,用解剖刀轻轻刮去内面组织,通过扫描电镜观察内表皮的表面结构。结果显示,蜕皮后1.5和3.0 h,头胸甲只包含上表皮和外表皮,KK-42处理组外表皮的板层数量显著增加,外表皮厚度分别比相应对照组提高了72.07%和38.67%;蜕皮后6.0~12.0 h,外表皮中疏松的板层趋于致密,其厚度在二组间无统计学差异。蜕皮后6.0 h,只有1层板层的内表皮出现,到蜕皮后12.0 h,板层数增至2层(对照组)和3层(处理组),且板层结构均疏松。此外,内表皮表面分布着大小不等、头尾走向的梭形孔道(pore canals);KK-42处理组内表皮结构未见明显变化。实验表明,KK-42对蜕皮后期日本沼虾幼虾头胸甲的超微结构有显著影响,能加快外表皮及内表皮板层的形成速率。     

Effects of cholesterol on growth,feed utilization,body composition and immune parameters in juvenile oriental river prawn,Macrobrachium nipponense (De Haan)

An 8‐week feeding trial was conducted to investigate the effects of dietary cholesterol levels on growth, feed utilization, body composition and immune parameters in juvenile oriental river prawn, Macrobrachium nipponense. Six isolipid (80 g kg^?1 crude lipid) and isoproteic (400 g kg^?1 crude protein) diets, supplemented with 0, 3.0, 6.0, 9.0, 12.0 and 15.0 g kg^?1 cholesterol, were evaluated. Growth performance and feed utilization of M. nipponense were improved as dietary cholesterol levels increased. Weight gain and specific growth rate were highest, and feed conversation ratio was lowest, when prawns were fed a diet supplemented with 9.0 g kg^?1 cholesterol. However, final body weights and survival rates of juvenile M. nipponense were not affected significantly by dietary cholesterol. Body composition of prawns, including moisture, crude protein and crude lipid, was not significantly affected by changes in dietary cholesterol. The immune parameters measured in hepatopancreas, including total antioxidant capacity, and glutathione, catalase, alkaline phosphatase and acid phosphatase activities, were at optimum levels in prawns fed with 9.0 g kg^?1 dietary cholesterol. In summary, the best growth performance, lowest feed conversation ratio, and the most enhanced antioxidant capacity and immunity parameters were attained in juvenile M. nipponense when fed a diet supplemented with 9.0 g kg^?1 cholesterol.     

Combined diet of yeast,fermented soybean meal,and microparticulate as larval feed in extensive rearing systems for seed production of the oriental river prawn <Emphasis Type="Italic">Macrobrachium nipponense</Emphasis>

The present study aims to evaluate the effects of several types of larval feed on seed production of the oriental river prawn Macrobrachium nipponense and water quality in extensive rearing systems. Healthy M. nipponense were distributed into fifteen 15-m² pond outdoors, and each pond was stocked with 90 prawns including 41 egg-bearing (berried) female prawns, 37 not bearing female prawns, and 12 male prawns for larval production. The experimental design was completely randomized, with four diet groups (diet A: soybean homogenate, diet B: yeast, diet C: yeast?+?fermented soybean meal, and diet D: yeast?+?fermented soybean meal?+?microparticulate diet) and one no feeding control and three replications. There are equal levels of the total protein content in diets A–D. Higher larvae and postlarvae number was obtained for diet C and diet D. Furthermore, the increase of M. nipponense larvae and postlarvae in diets B–D can result in the decrease of zooplankton and increase of phytoplankton in the pond during some periods, simultaneously improving the water quality. The Spearman correlation coefficient (r_M,N) between the amounts of larvae and postlarvae of M. nipponense per pond and zooplankton per pond during the entire experiment period was ??0.56. The highest juvenile M. nipponense production was obtained in the diet D group (3140.00?±?461.63), which was slightly higher than that in the diet C group (2940.33?±?382.60). The juvenile M. nipponense production in diet B (2303.00?±?449.23) was also significantly higher compared to diet A (1106.33?±?176.64) and the control group (1039.67?±?212.85). There were no significant differences in the amounts of juvenile M. nipponense between the control and diet A groups, which indicated that the soybean homogenate could not increase the amount of M. nipponense juveniles. Our results clearly showed that diet D significantly raised per unit yield of prawn seed comparing with no feeding and other diet group in extensive rearing systems and that the cost of producing prawn seed with diet D is low, which provide an important foundation for the expansion of oriental river prawn culture.     

Developmental changes in structure and polypeptide profile of the androgenic gland of the freshwater prawn Macrobrachium rosenbergii

Although the mechanism of sex-differentiation in crustaceans has yet to be defined, the androgenic gland (AG) is thought to be the exclusive organ that produces the androgenic hormone (AH) which induces male sexual development. This paper presents results of light and transmission electron microscopy and total protein analysis of androgenic glands from three male morphotypes (orange-claw, orange-blue-claw and blue-claw) of the freshwater prawn, Macrobrachium rosenbergii. Highest protein content (76 µg//AG) was found in the blue-claw morphotype as compared to the orange-blue-claw (45 µg/AG) and the orange-claw morphotype (19 µg/AG). Sodiumdodecyl sulfate polyacrylamide gel electrophoresis (SDS-page) analysis of the cell free extract of the AG from the three morphotypes revealed four polypeptides (16, 18, 23 and 26 Kd) which quantitatively increase from the sexually immature orange-claw to the sexually mature blue-claw morphotype. The 16 and 18 Kd polypeptides could be the AHs.