Science.gov

Sample records for macrobrachium rosenbergii nodavirus

  1. Interaction study of a novel Macrobrachium rosenbergii effector caspase with B2 and capsid proteins of M. rosenbergii nodavirus reveals their roles in apoptosis.

    PubMed

    Youngcharoen, Supak; Senapin, Saengchan; Lertwimol, Tareerat; Longyant, Siwaporn; Sithigorngul, Paisarn; Flegel, Timothy W; Chaivisuthangkura, Parin

    2015-08-01

    Apoptosis is an essential immune response to protect invertebrates from virus infected cells. In shrimp, virus infection has been reported to induce apoptosis. Macrobrachium rosenbergii (Mr) was considered to be a disease-resistant host when compared to penaeid shrimps. Caspase-3 was classified as an executioner caspase which played a key role in virus-induced apoptosis. In this study, an effector caspase gene of M. rosenbergii (Mrcasp) was cloned and characterized. The open reading frame (ORF) of Mrcasp was 957 nucleotide encoding 318 amino acid with a deduced molecular mass of 35.87 kDa. RT-PCR analysis showed the presence of Mrcasp in all examined tissues. The phylogenetic tree indicated that Mrcasp was closely related with caspase 3 of shrimp. The functions of the Mrcasp, B2 and capsid proteins of M. rosenbergii nodavirus (MrNV) were assayed in Sf-9 cells. The results showed that Mrcasp induce apoptotic morphology cells; however, capsid protein of MrNV could inhibit apoptotic cells whereas B2 could neither induce nor inhibit apoptotic cells by DAPI staining. The protein interaction between Mrcasp and viral MrNV structure revealed that Mrcasp did not bind to B2 or capsid protein whereas B2 and capsid proteins could bind directly to each other. This study reported a novel sequence of a full-length Mrcasp and its functional studies indicated that Mrcasp could induce apoptotic cells. Our data is the first report demonstrating the direct protein-protein interaction between capsid protein and B2 protein of MrNV. PMID:25982399

  2. Toxicity of rotenone to giant river freshwater prawn Macrobrachium rosenbergii

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aquaculturists have often suffered predation losses in the production of freshwater giant river prawn Macrobrachium rosenbergii due to the presence of wild fish species in culture ponds. The piscicide rotenone is widely used to remove undesirable fish species from ponds. Although evidence in the t...

  3. Ovarian and hemolymph ecdysteroid titers during vitellogenesis in Macrobrachium rosenbergii.

    PubMed

    Young, N J; Webster, S G; Rees, H H

    1993-05-01

    Changes in ovarian and hemolymph ecdysteroid concentration and composition during vitellogenesis have been investigated in the freshwater prawn Macrobrachium rosenbergii. Free ecdysteroids (20-hydroxyecdysone and ecdysone) in hemolymph increased in concentration during vitellogenesis from zero at stage 0 to 1.5 ng/ml at stage I to 7.3 ng/ml in mature, stage IV animals. 20-Hydroxyecdysonoic acid (1.2 ng/ml) was detected in the stage IV hemolymph. Ovarian-free ecdysteroid concentration, expressed as nanograms per gram of tissue, fell during vitellogenesis from 83.2 ng/g at stage 0, non-pigmented tissue to 14.2 ng/g at stage IV tissue, being minimal at stage I (6.3 ng/g). However, expression of ovarian free ecdysteroid content as nanograms per ovary revealed a rise from 7.7 ng/ovary at stage 0, nonpigmented tissue to 28.3 ng/ovary at stage IV, again being minimal at stage I (2.0 ng/ovary). 20-Hydroxyecdysonoic acid and ecdysonoic acid were identified at ovarian stages II-III and stage IV. PMID:8319875

  4. Grooming as a secondary behavior in the shrimp Macrobrachium rosenbergii (Crustacea, Decapoda, Caridea)

    PubMed Central

    VanMaurik, Lauren N.; Wortham, Jennifer L.

    2014-01-01

    Abstract The giant freshwater prawn, Macrobrachium rosenbergii, is a large shrimp extensively used in aquaculture whose grooming behaviors were analyzed in this study. Macrobrachium rosenbergii exhibits three unique male morphotypes that differ in their behavior, morphology and physiology: small-clawed males (SM), orange-clawed males (OC) and blue-clawed males (BC). The largest and most dominant males, BC males, are predicted to have significantly different grooming behaviors compared to females and the other two male morphotypes. These BC males may be too large and bulky to efficiently groom and may dedicate more time to mating and agonistic interactions than grooming behaviors. Observations were conducted to look at the prevalence of grooming behaviors in the absence and presence of conspecifics and to determine if any differences in grooming behavior exist among the sexes and male morphotypes. Significant differences in the grooming behaviors of all individuals (females and male morphotypes) were found. BC males tended to have the highest grooming time budget (percent of time spent grooming) while SM males had a relatively low grooming time budget. The grooming behaviors of the male morphotypes differed, indicating while these males play distinct, separate roles in the social hierarchy, they also have different grooming priorities. The conditions in which Macrobrachium rosenbergii are cultured may result in increased body fouling, which may vary, depending on the grooming efficiencies and priorities of these male morphotypes. Overall, grooming behaviors were found to be a secondary behavior which only occurred when primary behaviors such as mating, feeding or fighting were not present. PMID:25561831

  5. Potential use of the Macrobrachium rosenbergii lectin for diagnosis of T-cell acute lymphoblastic leukemia.

    PubMed

    Pérez-Campos-Mayoral, Laura; Ruiz-Argüelles, Alejandro; Pérez-Romano, Beatriz; Zenteno, Edgar; Hernández-Cruz, Pedro; Martínez-Cruz, Ruth; Martínez-Cruz, Margarito; Pina-Canseco, Socorro; Pérez-Campos, Eduardo

    2008-01-01

    T-cell acute lymphoblastic leukemia is the most common form of cancer in children. Lectins are proteins or glycoproteins from plants or animals that recognize oligossacharides on the cell surface and have been used to characterize the structural changes of oligosaccharides in leukemias. In this study, we used the lectin from the freshwater prawn Macrobrachium (M. rosenbergii), specific for acetyl groups in sialylated glycans, because increased sialylation of glycoproteins and glycolipids has been identified in lymphoblastic leukemias. We compared the specificity of the M. rosenbergii lectin for lymphoblastic leukemias with the specificities of the lectins from Triticum vulgaris, Solanum tuberosum, Arachis hipogaea, and Phytolacca americana. By morphologic and phenotype characterization with a panel of monoclonal antibodies, we identified four types of leukemias from 106 leukemia patients: 11 cases of T-cell acute lymphoblastic leukemia, 61 cases of B-cell acute lymphoblastic leukemia, 24 cases of acute myeloblastic leukemia, and 10 cases of acute biphenotypic leukemia. As determined by cytofluorometric assays, nine of the eleven cases with T-cell acute lymphoblastic leukemia (8 +/- 3 years old) were specifically identified with the lectin from M. rosenbergii. In contrast, only six cases of B-cell leukemia, one case of myeloblastic leukemia, and 2 cases of biphenotypic leukemia were identified with this M. rosenbergii lectin. The other lectins tested showed no capacity to differentiate, in a significant manner, any of the four types of leukemias tested. Thus, the lectin from M. rosenbergii could be considered a useful tool for the diagnosis and study of T-cell acute lymphoblastic leukemia. PMID:18212483

  6. TBT Effects on the Development of Intersex (Ovotestis) in Female Fresh Water Prawn Macrobrachium rosenbergii

    PubMed Central

    Peranandam, Revathi; Palanisamy, Iyapparaj; Lourdaraj, Arockia Vasanthi; Natesan, Munuswamy; Vimalananthan, Arun Prasanna; Thangaiyan, Suganya; Perumal, Anantharaman; Muthukalingan, Krishnan

    2014-01-01

    The impact of tributyltin (TBT) on the female gonad and the endocrine system in Macrobrachium rosenbergii was studied. Prawns were exposed to environmentally realistic concentrations of 10, 100, and 1000?ng/L of TBT for 6 months. Dose dependent effects were noticed in TBT exposed prawns. At 1000?ng/L TBT caused ovotestis formation (formation of male germ cells in ovary). Presence immature oocytes, fusion of developing oocytes, increase in interstitial connective tissues, and its modification into tubular like structure and abundance of spermatogonia in the ovary of TBT treated prawns. The control prawn ovary showed normal architecture of cellular organelles such as mature oocytes with type 2 yolk globules, lipid droplets, normal appearance of yolk envelop, and uniformly arranged microvilli. On the other hand, type 1 yolk globules, reduced size of microvilli, spermatogonial cells in ovary, spermatogonia with centrally located nucleus, and chromatin distribution throughout the nucleoplasm were present in the TBT treated group. Immunofluorescence staining indicated a reduction in vitellin content in ovary of TBT treated prawn. Moreover, TBT had inhibited the vitellogenesis by causing hormonal imbalance in M. rosenbergii. Thus, the present investigation demonstrates that TBT substantially affects sexual differentiation and gonadal development in M. rosenbergii. PMID:25121096

  7. Effect of cadmium on the ovarian development in the freshwater prawn Macrobrachium rosenbergii (De Man).

    PubMed

    Revathi, Peranandam; Vasanthi, Lourduraj Arockia; Munuswamy, Natesan

    2011-05-01

    In this investigation, effect of cadmium chloride (25 ?g/l) on oogenesis of freshwater prawn, Macrobrachium rosenbergii was studied. In vivo experiments were performed with both intact and eyestalk ablated prawns. The intact, cadmium-exposed prawns exhibited decrease in Gonado Somatic Index (GSI) and ovarian development compared to controls. Whereas, ablated treated ovary showed reduction of yolk material and oocyte membrane thickness at the end of 15 days exposure. Interestingly, the control prawn showed normal cellular architecture of gills, hepatopancreas and ovary with mature oocytes. But, the gills of treated prawns showed lamellar hypertrophy, cuticular dystrophy and irregular arrangement of epithelial cells. Hepatopancreas showed reduction in both tubular diameter and basement membrane thickness. Conspicuously, ovary showed hypertrophied primary oocytes with more vacuoles in intact-treated group. Cadmium had increased gonad inhibiting hormone (GIH) secretion and decreased gonad stimulating hormone (GSH) release as evident with the retardation of gonadal maturation in the intact prawns. PMID:21296420

  8. Impact of tributyltin on antioxidant and DNA damage response in spermatozoa of freshwater prawn Macrobrachium rosenbergii.

    PubMed

    Rani, K Umaa; Musthafa, M Saiyad; War, Mehrajuddin; Al-Sadoon, Mohammad K; Paray, Bilal Ahmad; Shareef, T H Mohamed Ahadhu; Nawas, P Mohideen Askar

    2015-12-01

    Effects of tributyltin (TBT) on antioxidant [total superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione reductase (GR)] and DNA damage levels in the spermatozoa were studied and reported here for the first time in the freshwater prawn Macrobrachium rosenbergii. Three groups of (n?=?10 in each group) fishes were exposed to three different nominal concentrations of TBT viz., 1, 2, and 4 mg L(-1) along with control group for 90 days. Significant decrease of antioxidant and increased DNA damage levels were seen at higher doses of 2 and 4 mg L(-1). In prawn, the antioxidant level plays a vital role in sperm protection, activation, differential functions related to the physiology, and reproductive behavior. This study serves as a biomonitoring tool to assess the TBT effects on reproductive behavior of aquatic biota. PMID:26296505

  9. Toxicity of phenol on Macrobrachium rosenbergii (de Man) eggs, larvae, and post-larvae

    SciTech Connect

    Law, A.T.; Yeo, M.E.

    1997-03-01

    Literature on the toxicities of phenol on aquatic organisms is very limited. USEPA reported that the acute and chronic toxicities of phenol to freshwater aquatic life occur at concentrations as low as 10.2 mg/L and 2.56 mg/L, respectively. While for the saltwater aquatic life the acute toxicity occurs at concentrations as low as 5.8 mg/L. No data are available for the chronic toxicity of phenol to saltwater aquatic life. Sublethal concentrations of phenol have significant effects on the physiological and histological processes of the aquatic organisms: such as gill necrosis; destruction of erythrocyte cells; inhibition of sexual activities; suppression on growth and reduction of resistance to diseases. Macrobrachium rosenbergii(de Man) is the sole freshwater prawn cultured in Malaysia. Occasionally, the hatcheries are unable to produce the post-larvae because of undefined pollutants present in the water supplies. It has been observed that the use of cracked fiberglass tanks for larvae rearing is correlated with high mortality. This high mortality is probably due to the toxicity of the phenolic compounds which are leached out from the fiber glass tank into the water. This study was undertaken to evaluate the toxicity of phenol on eggs, larvae and post-larvae of M. rosenbergii and to set the water quality criteria of phenol for the said species. 16 refs., 3 tabs.

  10. Involvement of Relish gene from Macrobrachium rosenbergii in the expression of anti-microbial peptides.

    PubMed

    Shi, Yan-Ru; Jin, Min; Ma, Fu-Tong; Huang, Ying; Huang, Xin; Feng, Jin-Ling; Zhao, Ling-Ling; Chen, Yi-Hong; Ren, Qian

    2015-10-01

    Relish is an NF-kB transcription factor involved in immune-deficiency (IMD) signal pathway. In this study, a Relish gene (MrRelish) was identified from Macrobrachium rosenbergii. The full length of MrRelish comprises 5072 bp, including a 3510 bp open reading frame encoding a 1169 bp amino acid protein. MrRelish contains a Rel homology domain (RHD), a nucleus localization signal, an I?B-like domain (6 ankyrin repeats), and a death domain. Phylogenetic analysis showed that MrRelish and other Relish from crustaceans belong to one group. MrRelish was expressed in all detected tissues, with the highest expression level in hemocytes and intestines. MrRelish was also upregulated in hepatopancreas at 6?h after Vibrio anguillarum challenge. The over-expression of MrRelish could induce the expression of antimicrobial peptides (AMPs), such as Drosophila Metchnikowin (Mtk), Attacin (Atta), Drosomycin (Drs), and Cecropin (CecA) and shrimp Penaeidin (Pen4). The RNAi of MrRelish in gills showed that the expression of crustin (cru) 2, Cru5, Cru8, lysozyme (Lyso) 1, and Lyso2 was inhibited. However, the expression of anti-lipopolysaccharide factor (ALF) 1 and ALF3 did not change when MrRelish was knocked down. These results indicate that MrRelish may play an important role in innate immune defense against V. anguillarum in M. rosenbergii. PMID:26026243

  11. Ultrastructure of differentiating oocytes and vitellogenesis in the giant freshwater prawn, Macrobrachium rosenbergii (De Man).

    PubMed

    Soonklang, Nantawan; Wanichanon, Chaitip; Stewart, Michael J; Stewart, Praphaporn; Meeratana, Prasert; Hanna, Peter J; Sobhon, Prasert

    2012-10-01

    The ultrastructure of oogenesis in Macrobrachium rosenbergii, with reference to vitellogenesis, has not been reported. We used light and electron microscopy, as well as vitellin (Vn) purification and antibody production, to study the temporal and spatial production of Vn in the ovary by immunofluorescence. Histologically, the ovary is subdivided into cone-shaped ovarian pouches with a central core containing layers of oogonia. These divide to produce oocytes that migrate outwardly and differentiate into mature oocytes. During the course of differentiation, oocytes undergo modifications, including the rearrangement of nuclear chromatin, the accumulation of ribosomes, rough endoplasmic reticulum (RER), and lipid, and the formation of secretory and yolk granules, resulting in four stages. Ultrastructurally, early previtellogenic oocytes (Oc(1)) are characterized by the accumulation of new ribosomal aggregates, translocated from the nucleus. Late previtellogenic oocytes (Oc(2)) show nuclear heterochromatin with a "clock face" pattern, the presence of RER, and three types of secretory granules. Follicular cells occupy the intercellular spaces and surround the Oc(2). Early vitellogenic oocytes (Oc(3)) are larger, with nuclei containing predominantly decondensed euchromatin, and cytoplasm with yolk and secretory granules, and few lipid droplets. Late vitellogenic oocytes (Oc(4)) are characterized by completely euchromatic nuclei, an indistinct plasma membrane, yolk platelets and secretory granules, and abundant lipid. Vitellogenin (Vg) in ovaries of M. rosenbergii consist of two main bands at MW 90 and 102 kDa. Our data indicates that Vn is present, and probably synthesized in Oc(3) and Oc(4), but there may be some undetected exogenous Vg production. PMID:22648947

  12. Transcriptomics of a Giant Freshwater Prawn (Macrobrachium rosenbergii): De Novo Assembly, Annotation and Marker Discovery

    PubMed Central

    Jung, Hyungtaek; Lyons, Russell E.; Dinh, Hung; Hurwood, David A.; McWilliam, Sean; Mather, Peter B.

    2011-01-01

    Background Giant freshwater prawn (Macrobrachium rosenbergii or GFP), is the most economically important freshwater crustacean species. However, as little is known about its genome, 454 pyrosequencing of cDNA was undertaken to characterise its transcriptome and identify genes important for growth. Methodology and Principal Findings A collection of 787,731 sequence reads (244.37 Mb) obtained from 454 pyrosequencing analysis of cDNA prepared from muscle, ovary and testis tissues taken from 18 adult prawns was assembled into 123,534 expressed sequence tags (ESTs). Of these, 46% of the 8,411 contigs and 19% of 115,123 singletons possessed high similarity to sequences in the GenBank non-redundant database, with most significant (E value < 1e–5) contig (80%) and singleton (84%) matches occurring with crustacean and insect sequences. KEGG analysis of the contig open reading frames identified putative members of several biological pathways potentially important for growth. The top InterProScan domains detected included RNA recognition motifs, serine/threonine-protein kinase-like domains, actin-like families, and zinc finger domains. Transcripts derived from genes such as actin, myosin heavy and light chain, tropomyosin and troponin with fundamental roles in muscle development and construction were abundant. Amongst the contigs, 834 single nucleotide polymorphisms, 1198 indels and 658 simple sequence repeats motifs were also identified. Conclusions The M. rosenbergii transcriptome data reported here should provide an invaluable resource for improving our understanding of this species' genome structure and biology. The data will also instruct future functional studies to manipulate or select for genes influencing growth that should find practical applications in aquaculture breeding programs. PMID:22174756

  13. Identification and characterization of the vitellogenin receptor in Macrobrachium rosenbergii and its expression during vitellogenesis.

    PubMed

    Roth, Ziv; Khalaila, Isam

    2012-07-01

    In oviparous organisms, oocyte maturation depends on massive production of the egg yolk-precursor protein, vitellogenin (Vg). Vg is taken up by the developing oocytes through receptor-mediated endocytosis (RME), a process essential to successful reproduction. The aims of this study were to identify and characterize the yet-unknown vitellogenin receptor (VgR) from the pleocyamate crustacean Macrobrachium rosenbergii, and to investigate its expression levels during vitellogenesis and its interaction with Vg. The VgR gene was cloned, and its translated protein was specifically located at the oocyte membrane. Moreover, for the first time, a VgR protein was identified and sequenced by mass spectrometry. The putative MrVgR displayed high sequence similarity to VgRs from crustaceans, insects, and vertebrates, and its structure includes typical elements, such as an extracellular, lipoprotein-binding domain (LBD), EGF-like, and O-glycosylation domains, a transmembrane domain, and a short, C-terminal, cytosolic tail. In this article, we identify the first crustacean VgR protein, and present data demonstrating its high affinity for a Vg column followed by elution with suramin and EDTA. Additionally we demonstrate that VgR expression in the oocyte is elevated during vitellogenesis. Our results contribute to the fundamental understanding of oocyte maturation in crustaceans, and particularly elucidate Vg uptake through RME via the VgR. PMID:22674884

  14. Studies on diseased freshwater prawn Macrobrachium rosenbergii infected with Vibrio vulnificus.

    PubMed

    Sharshar, Kh M; Azab, E A

    2008-09-01

    The present study was aimed at isolation and characterization of the pathogenic bacterium from diseased freshwater prawn. The effect of the bacterial pathogen on hepatopancreas, gills and exoskeleton was also investigated. Diseased freshwater prawn, Macrobrachium rosenbergii were collected from commercial hatchery in Behera Governorate, Egypt. The diseased prawn showed dark brown focal lesions and necrosis of appendage tips. The causative bacterial pathogen was isolated from haemolymph and hepatopancreas of the diseased prawn. Based on the morphological, biochemical and physiological characteristics, in addition EPI 20E test, the isolated pathogen was characterized as Vibrio vulnificus. Histopathology, hepatopancreas showed haemocytic infiltration in the interstitial sinuses, thickening and ruptures of the basal lamina and necrosis of its tubules. Similarly, the accumulation of haemocytes in the haemocoelic space, swelling, fusion of lamellae and abnormal gill tips. Also, the cuticular layers of the exoskeleton of diseased prawn had a rough or wrinkled surface and were disrupted and separated from the epidermis. The pathogen, V. vulnificus showed different degrees of sensitivity to different antimicrobial agents. It was highly sensitive to each of the antibiotics rifadin, virbamycin, oflaxcin, garamycin, flummox and trimethoprim/sulfamethoxzole) and resistant to nalidixic acid, unasyn, velosef, claforan, negram and amoxicillin. The minimal inhibitory concentration of trimethoprin/sulfamethoxzole for the studied pathogen, V. vulnificus was 0.31/5.93 microg. PMID:19266922

  15. Impact of TBT on the vitellogenesis and sex hormones in freshwater prawn Macrobrachium rosenbergii (De Man, 1879)

    PubMed Central

    2013-01-01

    Background Tributyltin (TBT) is a ubiquitous persistent xenobiotic that can be found in freshwater, estuarine and marine ecosystem. TBT is a strong endocrine disrupting compound (EDC) that can cause toxic threat to aquatic organisms. Imposex, sexual deformities and endocrine dysfunctions are the causes of TBT to most of the aquatic organisms. Effect of TBT on the vitellogenesis and sex hormonal changes in Macrobrachium rosenbergii has never been reported. Hence, the present investigation was undertaken to find out the impact of TBT on histological changes in the different reproductive tissues, sex hormonal alterations and level of biomarkers like vitellogenin and vitellin in M. rosenbergii. Results The present investigation documents the possible impact of tributyltin (TBT) on the vitellogenesis in freshwater female prawn M. rosenbergii. TBT at 10 ng/l, 100 ng/l and 1000 ng/l concentrations were exposed individually to prawns for a period of three months. At higher concentration of 1000 ng/l, the ovarian development was arrested and ovary remained at spent stage. At lower concentration of TBT (10 ng/l), the development proceeded up to early vitellogenic stage. At intermediate concentration of 100 ng/l TBT, the ovary remained at pre vitellogenic stage and thereafter no development was noticed. Histological results indicated the normal ovarian development with vitellogenic oocytes, filled with yolk globules in control prawn. On the other hand, the TBT treated groups showed reduction in yolk globules, fusion of developing oocytes and abundance of immature oocytes. Immunofluorescence staining denoted the remarkable reduction in vitellin content in ovary of TBT treated prawn. Hence, TBT had conspicuously inhibited the vitellogenesis by causing hormonal imbalance in M. rosenbergii. Conclusion TBT had notably inhibited the vitellogenesis due to hormonal imbalance. This endocrine dysfunction ultimately impaired the oogenesis in the freshwater female prawn M. rosenbergii. PMID:23634699

  16. Identification of the major allergen of Macrobrachium rosenbergii (giant freshwater prawn)

    PubMed Central

    Yadzir, Zailatul Hani Mohamad; Misnan, Rosmilah; Abdullah, Noormalin; Bakhtiar, Faizal; Arip, Masita; Murad, Shahnaz

    2012-01-01

    Objective To characterize the major allergens of Macrobrachium rosenbergii (giant freshwater prawn). Methods Raw and cooked extracts of the giant freshwater prawn were prepared. The IgE reactivity pattern was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting technique with the sera of 20 skin prick test (SPT) positive patients. The major allergen identified was then characterized using the proteomics approach involving a combination of two-dimensional (2-DE) electrophoresis, mass spectrometry and bioinformatics tools. Results SDS-PAGE of the raw extract showed 23 protein bands (15–250 kDa) but those ranging from 40 to 100 kDa were not found in the cooked extract. From immunoblotting experiments, raw and cooked extracts demonstrated 11 and 5 IgE-binding proteins, respectively, with a molecular mass ranging from 15 to 155 kDa. A heat-resistant 36 kDa protein was identified as the major allergen of both extracts. In addition, a 42 kDa heat-sensitive protein was shown to be a major allergen of the raw extract. The 2-DE gel fractionated the prawn proteins to more than 50 different protein spots. Of these, 10 spots showed specific IgE reactivity with patients' sera. Matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis led to identification of 2 important allergens, tropomyosin and arginine kinase. Conclusions It can be concluded that the availability of such allergens would help in component-based diagnosis and therapy of prawn allergies. PMID:23569834

  17. Molecular cloning and characterization of a Toll receptor gene from Macrobrachium rosenbergii.

    PubMed

    Srisuk, Chutima; Longyant, Siwaporn; Senapin, Saengchan; Sithigorngul, Paisarn; Chaivisuthangkura, Parin

    2014-02-01

    Toll receptors are cell surface molecules acting as pattern recognition receptors (PRRs) that have been implicated in the signaling pathway of innate immune responses. In this study, the full-length cDNA of a Toll receptor gene of Macrobrachium rosenbergii, designated MrToll, was successfully isolated using designed degenerate primers and the rapid amplification of cDNA ends (RACE). The MrToll gene sequence contained an open reading frame (ORF) of 2799 nucleotides encoding a protein of 932 amino acid residues. The protein contained distinct structural motifs of the Toll-like receptor (TLR) family, including an extracellular domain containing 15 leucine-rich repeats (LRRs), a transmembrane segment of 23 amino acids, and a cytoplasmic Toll/interleukin-1R (TIR) domain of 139 residues. Phylogenetic analysis revealed that MrToll and Toll receptor of Marsupenaeus japonicus (MjToll) evolved closely. However, the MrToll ORF demonstrated only 48-49% identity with shrimp Toll1, suggesting that MrToll isolated from a palaemonid shrimp might belong to a novel class of Toll receptors in shrimp. The transcripts of the MrToll gene were constitutively expressed in various tissues, with high levels in hemocytes, the stomach and muscle. A reverse transcriptase PCR assay demonstrated that the expression patterns of MrToll were distinctly modulated after Aeromonas caviae stimulation, with significant enhancement at 3-12 h post-challenge and a decline to basal levels at 24 h post-challenge. In addition, when MrToll-silenced shrimp were challenged with A. caviae, there was a significant increase in mortality and bacterial CFU counts. These results suggest that MrToll might be involved in host innate defense, especially against the pathogen A. caviae. PMID:24398262

  18. In silico Neuropeptidome of Female Macrobrachium rosenbergii Based on Transcriptome and Peptide Mining of Eyestalk, Central Nervous System and Ovary

    PubMed Central

    Wang, Tianfang; Zhao, Min; Elizur, Abigail; Sretarugsa, Prapee; Cummins, Scott F.; Sobhon, Prasert

    2015-01-01

    Macrobrachium rosenbergii is the most economically important of the cultured freshwater crustacean species, yet there is currently a deficiency in genomic and transcriptomic information for research requirements. In this study, we present an in silico analysis of neuropeptide genes within the female M. rosenbergii eyestalk, central nervous system, and ovary. We could confidently predict 37 preproneuropeptide transcripts, including those that encode bursicons, crustacean cardioactive peptide, crustacean hyperglycemic hormones, eclosion hormone, pigment-dispersing hormones, diuretic hormones, neuropeptide F, neuroparsins, SIFamide, and sulfakinin. These transcripts are most prominent within the eyestalk and central nervous system. Transcript tissue distribution as determined by reverse transcription-polymerase chain reaction revealed the presence of selected neuropeptide genes of interest mainly in the nervous tissues while others were additionally present in the non-nervous tissues. Liquid chromatography-mass spectrometry analysis of eyestalk peptides confirmed the presence of the crustacean hyperglycemic hormone precursor. This data set provides a strong foundation for further studies into the functional roles of neuropeptides in M. rosenbergii, and will be especially helpful for developing methods to improve crustacean aquaculture. PMID:26023789

  19. Carbon, nitrogen, and phosphorus budget in scampi (Macrobrachium rosenbergii) culture ponds.

    PubMed

    Sahu, Bharat Chandra; Adhikari, Subhendu; Mahapatra, Abhijit S; Dey, Lambodar

    2013-12-01

    Experiments were conducted for the study of nutrient budget in ten farmer's ponds (0.2-0.5 ha) in Orissa, India with a mean water depth of 1.0-1.2 m. Scampi (Macrobrachium rosenbergii) were stocked in these ponds at stocking density of 3.75-5.0/m(2). The average initial body weight of scampi was 0.02 mg. The culture period was for 4 months. Feed was the main input. Total feed applied to these ponds ranged from 945 to 2261 kg pond/cycle (crop). The feed conversion ratio varied 1.65 to 1.78. In addition to feed, rice straw, urea, and single super phosphate were applied to these ponds in small amounts for plankton production. At harvest time, the average weight of scampi varied from 60-90 g. The budget showed that feed was the major input of nitrogen (N), phosphorus (P), and carbon in these ponds. The inorganic fertilizer (urea and single super phosphate), organic fertilizer (rice straw and yeast extract), and inlet water, either from the initial fills or from rainwater, were the source of all other N, P, and organic carbon (OC) to these ponds. Total N applied to these ponds through all these inputs ranged from 44.45 to 103.98 kg N per crop, 12.23 to 28.79 kg P per crop, and from 381.54 to 905.22 kg OC per crop, respectively. Among all the inputs, feed alone accounted for 95.34 % N, 97.98 % P, and 94.27 % OC, respectively. Recovery of 16.34 to 38.66 kg N (average 29.27 kg), 1.28 to 3.02 kg P (average 2.29 kg), and 63.21 to 149.51 kg OC (average 113.20 kg), respectively, by the scampi harvest were observed in these ponds. Thus, harvest of scampi accounted for recovery of 35.18 to 39.01 (average 36.85%) of added N, 10.09 to 10.97 (average 10.44%) of added P, and 7.57 to 17.12 (average 16.34%) of added OC, respectively. PMID:23832231

  20. Spermatophore affects the egg-spawning and egg-carrying behavior in the female giant freshwater prawn, Macrobrachium rosenbergii.

    PubMed

    Kruangkum, Thanapong; Vanichviriyakit, Rapeepun; Chotwiwatthanakun, Charoonroj; Saetan, Jirawat; Tinikul, Yotsawan; Wanichanon, Chaitip; Cummins, Scott F; Hanna, Peter J; Sobhon, Prasert

    2015-10-01

    In crustaceans, mating occurs during the ecdysis after female molting. During this period, a male transfers its spermatophore into a female which, in some species, stores the spermatophore for a long period prior to spawning and fertilization. However, in some species including the giant freshwater prawn, Macrobrachium rosenbergii, the male deposits its spermataphore onto the external surface of the thoracic segment of the female which affects the spawning time and maternal behavior. This study investigated the spawning behavior of the M. rosenbergii females, which was divided into pre-spawning, spawning, and post-spawning phases. It was revealed that mated female prawns with attached spermatophore exhibited an earlier spawning than unmated individuals, leading to assessment of the factors that may elicit this phenomenon. Four groups of female prawns were allocated to groups including mating females with spermatophore still attached, mating females with the spermatophore removed, artificially inseminated females with spermatophores, and an unmated control. There was a significant reduction in the time of egg-spawning in the presence of spermatophores, and the mating activity was also a contributing factor. Furthermore, over 90% of the mated and artificially inseminated females in which spermatophores were deposited carried the eggs in the abdominal brood chamber until completion of embryonic development while others discarded the eggs within 2 days post-spawning. This study implies that the spermatophore may contain ovulation-inducing factors which stimulate an earlier spawning and fostering of brooding behavior. PMID:26388033

  1. A first insight into temperature stress-induced neuroendocrine and immunological changes in giant freshwater prawn, Macrobrachium rosenbergii.

    PubMed

    Chang, Chin-Chyuan; Jiang, Jia-Rong; Cheng, Winton

    2015-11-01

    Haemolymph norepinephrine (NE); total haemocyte count (THC); respiratory bursts (RBs); superoxide dismutase (SOD), phenoloxidase (PO), and phagocytic activity; and prophenoloxidase (proPO)-system-related genes (lipopolysaccharide- and ?-1,3-glucan-binding protein: LGBP, proPO, peroxinectin: PE, and ?2-macroglobulin: ?2-M) in haemocytes of Macrobrachium rosenbergii were investigated after transferring them from 28 °C to 22 °C, 28 °C, and 34 °C respectively. The results revealed that haemolymph NE, hyaline cells (HCs), and PO activity per granulocyte increased from 30 to 120 min of exposure, and however, RBs and phagocytic activity significantly decreased from 30 to 120 min of exposure as well as granular cells (GCs), semigranular cells (SGCs), and SOD activity decreased from 60 to 120 min of exposure for the prawns subjected to temperature stress. The proPO-system-related gene expression markedly increased with 60-120 min of exposure for the prawns transferred from 28 °C to 22 °C and 34 °C, except ?2M at 120 min. These results provide a first insight into the effects of temperature stress on haemolymph NE level and immune functions in prawns and suggest that temperature-stress-induced acute modulation in immunity is associated with the release of haemolymph NE in M. rosenbergii. PMID:26434711

  2. Retardation of post-mortem changes of freshwater prawn (Macrobrachium rosenbergii) stored in ice by legume seed extracts.

    PubMed

    Sriket, Chodsana; Benjakul, Soottawat; Visessanguan, Wonnop; Hara, Kenji; Yoshida, Asami

    2012-11-15

    Meat quality of freshwater prawn (Macrobrachium rosenbergii) treated with soybean and bambara groundnut extracts at different concentrations was monitored during 10 days of iced storage. During storage, the control sample (without treatment) had a higher pH, TCA-soluble peptide content, heat soluble collagen content, proteolytic activities and psychrophilic bacterial count than did samples treated with soybean and bambara groundnut extracts. Conversely, shear force value and likeness scores of the control sample decreased (p<0.05), more likely associated with softening of muscle. The decrease in myosin heavy chain in the control sample was found after 6 days of storage. However, no changes in protein patterns of samples treated with soybean extracts at 2.5 mg/mL were found after 10 days of storage. Therefore, the injections of legume seed extracts, especially soybean extract, at a sufficient concentration, could be a means to retard muscle softening and maintain the qualities of freshwater prawn during iced storage. PMID:22868130

  3. Expression of biotransformation and oxidative stress genes in the giant freshwater prawn Macrobrachium rosenbergii exposed to chlordecone.

    PubMed

    Gaume, Béatrice; Dodet, Nathalie; Thomé, Jean-Pierre; Lemoine, Soazig

    2015-06-01

    Chlordecone is a persistent organochlorine pesticide widely used between 1972 and 1993 in the French West Indies to control the root borer in banana fields. Chlordecone use resulted in long-term pollution of soils, contamination of waters, of aquatic organisms, and of fields. Chlordecone is known to be neurotoxic, to increase prostate cancer, and to have negative effects on cognitive and motor development during infancy. In Guadeloupe, most of the freshwater species living in contaminated rivers exceed the French legal limit of 20 ?g·kg(-1) wet weight. In the present study, we chose a transcriptomic approach to study the cellular effects of chlordecone in the giant freshwater prawn Macrobrachium rosenbergii, an important economical species in Guadeloupe. Quantitative PCR revealed an induction of genes involved in defense mechanism against oxidative stress (catalase and selenium-dependent glutathione peroxidase) in prawns exposed to low environmental concentrations of chlordecone after 12 and 24 h of exposure. In prawns reared in a contaminated farm, transcription of genes involved in the biotransformation process (cytochrome P450 and glutathione-S-transferase (GST)) were induced after 8 days of exposure. Our results provide information on the mechanims of defense induced by chlordecone in aquatic crustacean species. This gene expression study of selected genes should be further strengthened by proteomic analyses and enzymatic activity assays to confirm the response of these biomarkers of stress in crustaceans and to give new insights into the mechanism of toxicity by chlordecone. PMID:24920261

  4. De Novo Transcriptome Sequencing Analysis and Comparison of Differentially Expressed Genes (DEGs) in Macrobrachium rosenbergii in China

    PubMed Central

    Liu, Qigen

    2014-01-01

    Giant freshwater prawn (GFP; Macrobrachium rosenbergii) is an exotic species that was introduced into China in 1976 and thereafter it became a major species in freshwater aquaculture. However the gene discovery in this species has been limited to small-scale data collection in China. We used the next generation sequencing technology for the experiment; the transcriptome was sequenced of samples of hepatopancreas organ in individuals from 4 GFP groups (A1, A2, B1 and B2). De novo transcriptome sequencing generated 66,953 isogenes. Using BLASTX to search the Non-redundant (NR), Search Tool for the Retrieval of Interacting Genes (STRING), and Kyoto Encyclopedia of Genes and Genome (KEGG) databases; 21,224 unigenes were annotated, 9,552 matched unigenes with the Gene Ontology (GO) classification; 5,782 matched unigenes in 25 categories of Clusters of Orthologous Groups of proteins (COG) and 20,859 unigenes were consequently assigned to 312 KEGG pathways. Between the A and B groups 147 differentially expressed genes (DEGs) were identified; between the A1 and A2 groups 6,860 DEGs were identified and between the B1 and B2 groups 5,229 DEGs were identified. After enrichment, the A and B groups identified 38 DEGs, but none of them were significantly enriched. The A1 and A2 groups identified 21,856 DEGs in three main categories based on functional groups: biological process, cellular_component and molecular function and the KEGG pathway defined 2,459 genes had a KEGG Ortholog - ID (KO-ID) and could be categorized into 251 pathways, of those, 9 pathways were significantly enriched. The B1 and B2 groups identified 5,940 DEGs in three main categories based on functional groups: biological process, cellular_component and molecular function, and the KEGG pathway defined 1,543 genes had a KO-ID and could be categorized into 240 pathways, of those, 2 pathways were significantly enriched. We investigated 99 queries (GO) which related to growth of GFP in 4 groups. After enrichment we identified 23 DEGs and 1 KEGG PATHWAY ‘ko04711’ relation with GFP growth. PMID:25329319

  5. Variation of prostaglandin E2 concentrations in ovaries and its effects on ovarian maturation and oocyte proliferation in the giant fresh water prawn, Macrobrachium rosenbergii.

    PubMed

    Sumpownon, Chanudporn; Engsusophon, Attakorn; Siangcham, Tanapan; Sugiyama, Eiji; Soonklang, Nantawan; Meeratana, Prasert; Wanichanon, Chaitip; Hanna, Peter J; Setou, Mitsutoshi; Sobhon, Prasert

    2015-11-01

    Prostaglandins (PGs) are important bioactive mediators for many physiological functions. In some decapod crustaceans, prostaglandin E2 (PGE2) has been detected in reproductive organs, and may play a role in the control of ovarian maturation. However, in the freshwater prawn, Macrobrachium rosenbergii, the presences of PGE2 and key enzymes for PGE2 biosynthesis, as well as its effects on ovarian maturation have not yet been investigated. In this study we reported the presence of PGE2, cyclooxygenase1 (COX1) and prostaglandin E synthase (PGES) in the ovarian tissues of M. rosenbergii, using immunohistochemistry. Intense immunoreactivities of PGE2 (PGE2-ir), COX1 (Cox1-ir) and PGES (PGES-ir) were detected in previtellogenic oocytes (Oc1 and Oc2), while the immunoreactivities were absent in the late vitellogenic oocytes (Oc4). This finding supports the hypothesis that the PGE2 biosynthesis occurs in the ovary of this prawn. To ascertain this finding we used LC-MS/MS to quantitate PGE2 concentrations during ovarian developmental cycle. The levels of PGE2 were significantly higher in the early ovarian stages (St I and II) than in the late stages (St III and IV). Moreover, we found that administration of PGE2 stimulated the ovarian maturation in this species by shortening the length of the ovarian cycle, increasing ovarian-somatic index, oocyte proliferation, and vitellogenin (Vg) level in the hemolymph. PMID:25963041

  6. Molecular cloning and characterisation of prophenoloxidase cDNA from haemocytes of the giant freshwater prawn, Macrobrachium rosenbergii, and its transcription in relation with the moult stage.

    PubMed

    Liu, Chun-Hung; Tseng, Deng-Yu; Lai, Ching-Yi; Cheng, Winton; Kuo, Ching-Ming

    2006-07-01

    Expression of prophenoloxidase (proPO) cDNA was determined from haemocytes of the giant freshwater prawn Macrobrachium rosenbergii by a reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA using oligonucleotide primers based on the proPO sequence of tiger shrimp Penaeus monodon, freshwater crayfish Pacifastacus leniusculus, green tiger shrimp Penaeus semisulcatus, kuruma shrimp Marsupenaeus japonicus, and white shrimp Litopenaeus vannamei. The proPO of M. rosenbergii was constitutively expressed. The 2,547-bp cDNA contained an open reading frame (ORF) of 2,013 bp, a 96-bp 5'-untranslated region, and a 438-bp 3'-untranslated region containing the poly A tail. The molecular mass of the deduced amino acid (aa) sequence (671 aa) was 76.7 kDa with an estimated pI of 7.05. It contained putative copper-binding sites, a complement-like motif (GCGWPRHM), a proteolytic activation site, and a conserved C-terminal region common to all known proPOs. However, no signal peptide sequence was detected in giant freshwater prawn proPO. Comparison of amino acid sequences showed that prawn proPO is similar to the proPO of penaeid, crayfish and lobster. Prawn proPO was only synthesised in haemocytes. The proPO transcript was significantly increased in the A stage and achieved the highest level in the B stage, and then declined sharply in the C stage and reached the lowest level in the D(2)/D(3) stage. PMID:16376571

  7. Low pH affects survival, growth, size distribution, and carapace quality of the postlarvae and early juveniles of the freshwater prawn Macrobrachium rosenbergii de Man

    NASA Astrophysics Data System (ADS)

    Kawamura, Gunzo; Bagarinao, Teodora; Yong, Annita Seok Kian; Chen, Chiau Yu; Noor, Siti Norasidah Mat; Lim, Leong Seng

    2015-06-01

    Acidification of rain water caused by air pollutants is now recognized as a serious threat to aquatic ecosystems. We examined the effects of low pH (control pH 7.5, pH 6, pH 5, pH 4) on the survival, growth, and shell quality of Macrobrachium rosenbergii postlarvae and early juveniles in the laboratory. Hatcheryproduced postlarvae (PL 5) were stocked at 250 PL per aquarium, acclimated over 7 d to experimental pH adjusted with hydrochloric acid, and reared for 30 d. Dead specimens were removed and counted twice a day. After 27 d rearing, all specimens were measured for total length and body weight. Carapace quality was assessed by spectrophotometry. Survival of juveniles was highest at pH 6 (binomial 95% confidence interval 79 - 89%) followed by control pH 7.5 (56 - 68%) and pH 5 (50 - 60%) and was lowest for unmetamorphosed postlarvae and juveniles at pH 4 (43 - 49%). The final median total length and body weight of juveniles were similar at control pH 7.5 (18.2 TL, 50.2 mg BW) and pH 6 (17.7 mm TL, 45.0 mg BW) but significantly less at pH 5 (16.7 mm TL, 38.2 mg BW); at pH 4, the postlarvae did not metamorphose and measured only 9.8 mm TL, 29.3 mg BW. Length frequency distribution showed homogeneous growth at pH 6, positive skew at control pH 7.5 and pH 5, and extreme heterogeneity at pH 4. The carapace showed different transmittance spectra and lower total transmittance (i.e. thicker carapace) in juveniles at pH 7.5, pH 6, and pH 5 than in unmetamorphosed postlarvae and juveniles with thinner carapace at pH 4. Thus, survival, growth, size distribution, and carapace quality of M. rosenbergii postlarvae and early juveniles were negatively affected by pH 5 and especially pH 4. The thinner carapace of the survivors at pH 4 was mostly due to their small size and failure to metamorphose. Natural waters affected by acid rain could decimate M. rosenbergii populations in the wild.

  8. L-( sup 3 H) glutamate binding to a membrane preparation from the optic lobe of the giant freshwater prawn Macrobrachium rosenbergii de Man

    SciTech Connect

    Pratumtan, P.; Govitrapong, P.; Withyachumnarnkul, B.; Poolsanguan, B. Mahidol Univ., Nakorn Pathom )

    1991-01-01

    Membrane preparation from the optic lobe of the giant freshwater prawn, Macrobrachium rosenbergii de Man, was examined for the presence of specific L-({sup 3}H) glutamate binding. The optic lobes were isolated from live animals. The tissue was homogenized and the membrane fraction isolated by differential centrifugation. The membrane suspension was incubated with 10-1,000 nM of L-({sup 3}H) glutamate at 37{degree}C for 60 min. Nonspecific binding was determined by incubating the mixture with 100 {mu}M L-glutamate. L-({sup 3}H) glutamate specifically bound to the membrane fraction with a dissociation equilibrium constant (Kd) of 205 nM and maximum number of binding sites (Bmax) of 2.04 n mol/mg protein. By using LIGAND computerized program, the saturation isotherm binding pattern indicates a single type of binding. To determine the type of glutamate receptors, competitive inhibition and IC{sub 50} of several glutamate agonists and antagonists were determined. The study reveals a metabotropic type of binding site.

  9. Characterization of a novel nm23 gene and its potential roles in gametogenesis in the prawn Macrobrachium rosenbergii (de Man, 1879) (Crustacea: Decapoda).

    PubMed

    Song, Ya-Nan; Lu, Cui-Yun; Chen, Jie; Qiu, Gao-Feng

    2013-11-15

    Nm23 is a family of genes encoding the nucleoside diphosphate (NDP) kinase, which functions in a wide variety of biological processes, including growth, development, differentiation and tumor metastasis. In this study, a novel nm23 gene, designated as Mrnm23, was identified from the freshwater giant prawn Macrobrachium rosenbergii. The full-length cDNA was 776bp in length, encoding for a protein of 176 amino acids with one typical NDP kinase domain that harbored all the crucial residues for nucleotide binding and enzymatic activity. Like human novel nm23-H1B, the putative protein contained a unique 21-amino-acid NH2-terminal extension as compared to human nm23 (nm23-H1) homologs. Further, 3 extra amino acid residues prolonged the COOH-terminus. The Mrnm23 was ubiquitously expressed in all tissues examined, including androgenic gland, gill, heart, liver, muscle, ovary, and testis. In situ hybridization to gonad sections indicated that the Mrnm23 mRNA was localized in the cytoplasm of cup-base of differentiating spermatids, in the spike of the umbrella-shaped spermatozoa and in the cytoplasm of the early previtellogenic oocytes, suggesting that the Mrnm23 has potential roles in spermiogenesis and early differentiation of oocyte. PMID:23994193

  10. Proteomic analysis of differentially expressed protein in hemocytes of wild giant freshwater prawn Macrobrachium rosenbergii infected with infectious hypodermal and hematopoietic necrosis virus (IHHNV)

    PubMed Central

    Alinejad, T.; Bin, Kwong Q.; Vejayan, J.; Othman, R.Y.; Bhassu, S.

    2015-01-01

    Epizootic diseases cause huge mortality and economical loses at post larvae stages in freshwater prawn aquaculture industry. These prawns seem less susceptible to viral diseases except for infectious hypodermal and hematopoietic necrosis virus (IHHNV). During viral infection in prawns, hemocytes are the primary organ that shows immunological response within the early stages of infection. We applied proteomic approaches to understand differential expression of the proteins in hemocytes during the viral disease outbreak. To aid the goal, we collected Macrobrachium rosenbergii broodstocks from the local grow out hatchery which reported the first incidence of IHHNV viral outbreak during larvae stage. Primarily, application of the OIE primer targeting 389 bp fragments of IHHNV virus was used in identification of the infected and non-infected samples of the prawn breeding line. Analysis of two-dimensional gel electrophoresis showed specific down-regulation of Arginine kinase and Sarcoplasmic calcium-binding protein and up/down-regulation of Prophenoloxidase1 and hemocyanin isoforms. These proteins were validated using semi quantitative RT-PCR and gene transcripts at mRNA level. These identified proteins can be used as biomarkers, providing a powerful approach to better understanding of the immunity pathway of viral disease with applications in analytic and observational epidemiology diagnosis. Proteomic profiling allows deep insight into the pathogenesis of IHHNV molecular regulation and mechanism of hemocyte in freshwater prawns. PMID:26106581

  11. Changes in the levels, expression, and possible roles of serotonin and dopamine during embryonic development in the giant freshwater prawn, Macrobrachium rosenbergii.

    PubMed

    Tinikul, Yotsawan; Poljaroen, Jaruwan; Tinikul, Ruchanok; Sobhon, Prasert

    2016-01-01

    We investigated the changes in the levels of serotonin (5-HT) and dopamine (DA), and their possible roles during embryonic development of the freshwater prawn, Macrobrachium rosenbergii. The 5-HT and DA concentrations were quantified using high performance liquid chromatography with electrochemical detection (HPLC-ECD). The levels of 5-HT and DA gradually increased from early developing embryos to late developing embryos. The 5-HT concentrations gradually increased from the pale yellow egg to orange egg stages, and reaching a maximum at the black egg stage. DA concentrations were much lower in the early embryos than those of 5-HT (P<0.05), and gradually increased to reach the highest level at the black egg stage. Immunohistochemically, 5-HT was firstly detected in the early embryonic stages, whereas DA developed later than 5-HT. Functionally, 5-HT-treated female prawns at doses of 2.5×10(-5), 2.5×10(-6) and 2.5×10(-7)mol/prawn, produced embryos with significantly shortened lengths of early embryonic stages, whereas DA-treated prawns at all three doses, exerted its effects by significantly lengthening the period of mid-embryonic stage onwards. These results suggest significant involvement of 5-HT and DA in embryonic developmental processes of this species. PMID:26393313

  12. Proteomic analysis of differentially expressed protein in hemocytes of wild giant freshwater prawn Macrobrachium rosenbergii infected with infectious hypodermal and hematopoietic necrosis virus (IHHNV).

    PubMed

    Alinejad, T; Bin, Kwong Q; Vejayan, J; Othman, R Y; Bhassu, S

    2015-09-01

    Epizootic diseases cause huge mortality and economical loses at post larvae stages in freshwater prawn aquaculture industry. These prawns seem less susceptible to viral diseases except for infectious hypodermal and hematopoietic necrosis virus (IHHNV). During viral infection in prawns, hemocytes are the primary organ that shows immunological response within the early stages of infection. We applied proteomic approaches to understand differential expression of the proteins in hemocytes during the viral disease outbreak. To aid the goal, we collected Macrobrachium rosenbergii broodstocks from the local grow out hatchery which reported the first incidence of IHHNV viral outbreak during larvae stage. Primarily, application of the OIE primer targeting 389 bp fragments of IHHNV virus was used in identification of the infected and non-infected samples of the prawn breeding line. Analysis of two-dimensional gel electrophoresis showed specific down-regulation of Arginine kinase and Sarcoplasmic calcium-binding protein and up/down-regulation of Prophenoloxidase1 and hemocyanin isoforms. These proteins were validated using semi quantitative RT-PCR and gene transcripts at mRNA level. These identified proteins can be used as biomarkers, providing a powerful approach to better understanding of the immunity pathway of viral disease with applications in analytic and observational epidemiology diagnosis. Proteomic profiling allows deep insight into the pathogenesis of IHHNV molecular regulation and mechanism of hemocyte in freshwater prawns. PMID:26106581

  13. The acute modulation of norepinephrine on immune responses and genes expressions via adrenergic receptors in the giant freshwater prawn, Macrobrachium rosenbergii.

    PubMed

    Chang, Chin-Chyuan; Tsai, Wan-Lin; Jiang, Jia-Rong; Cheng, Winton

    2015-10-01

    Norepinephrine (NE), immunocompetent parameters (total haemocyte count (THC), phenoloxidase (PO) activity, respiratory burst (RB), superoxide dismutase (SOD) activity, phagocytic activity and clearance efficiency to Lactococcus garvieae), and prophenoloxidase (proPO) system-related genes (lipopolysaccharide- and ?-1,3-glucan-binding protein, LGBP; prophenoloxidase, proPO; peroxinectin, PE; ?2-macroglobulin, ?2-M) expressions were investigated in Macrobrachium rosenbergii received NE through injection at 50 pmol/prawn after 0, 30, 60, and 120 min. Furthermore, the PO activity, RB, SOD activity, phagocytic activity and proPO system-related genes expressions were determined in haemocytes incubated with cacodylate buffer (CAC), NE, and NE co-treated with various adrenergic receptor (AR) antagonists in vitro. Results showed that NE, THC, granular cells, PO activity, SOD activity, proPO system-related genes expressions, and phagocytic activity and clearance efficiency to L. garvieae increased; PO activity per granulocyte and RB per haemocyte decreased from 30 to 120 min; semigranular cells and RB increased in the initial 30 min, and then decreased at 120 min when the prawns received NE by injection. In vitro studies, all the determined immune parameters and genes expressions were significantly decreased in haemocytes incubated with NE after 30 min. The negative effects of NE were prevented on the PO activity and phagocytic activity by the ?-AR antagonist of metoprolol (Met), on the SOD activity by the ?-AR antagonist of propranolol (Pro), on the RB by the ?-AR antagonist of Met and prazosin (Pra), and on the proPO system-related genes expressions by ?-AR antagonist of Pra. These results show that NE modulates prawn haemocytes proPO system-related genes expressions via ?1-AR, PO activity and phagocytosis via ?1-AR, respiratory burst via ?1-and ?1-ARs, and SOD activity via ?2-AR. It is concluded that NE stimulates the regulation of immunocompetence parameters and proPO system-related genes expressions in an acute response to maintain homeostasis of M. rosenbergii, which is primarily mediated through ?1-, ?1-and ?2-ARs. PMID:26210701

  14. Evaluation of paraprobiotic applicability of Lactobacillus plantarum in improving the immune response and disease protection in giant freshwater prawn, Macrobrachium rosenbergii (de Man, 1879).

    PubMed

    Dash, Gyanaranjan; Raman, Ram Prakash; Pani Prasad, K; Makesh, M; Pradeep, M A; Sen, Swatipriyanka

    2015-03-01

    Paraprobiotics, also known as ghost probiotics, are non-viable microbial cells which, when administered in adequate amounts, confer a benefit on the host. However, the advantage of non-viable microbes over their viable counterparts is a much debated topic in aquaculture. Therefore, the present study was conducted to evaluate paraprobiotic effect of heat-killed Lactobacillus plantarum on giant freshwater prawn Macrobrachium rosenbergii. A 90-day feeding trial was conducted by feeding prawn juveniles (mean weight ± SE: 0.54 ± 0.03 g) with three experimental diets prepared by supplementing basal diet (Crude protein: 38%; Gross energy: 387 kcal 100 g(-1)) with different concentrations of heat-killed probiotics bacteria viz. HKPB1 (10(7) cfu g(-1) diet), HKPB2 (10(8) cfu g(-1) diet), HKPB3 (10(9) cfu g(-1) diet) and control diet (unsupplemented diet). In the present study, growth parameters viz. WG % and SGR and feed utilization parameters viz. FCE, FCR and PER, though improved marginally in all experimental groups, were found to be insignificant (P > 0.05) compared to the control. The immune parameters viz. total hemocyte count (THC), phenol oxidase (PO) activity, respiratory burst (RB) activity and clearance efficiency were significantly improved (P < 0.05) with concurrent decrease (P < 0.05) in cumulative mortality against Aeromonas hydrophila challenge in all the experimental groups except for HKPB1, where PO and RB activity did not improve significantly (P > 0.05) compared to the control. Among the experimental groups, though the improvement in immune parameters was higher (P < 0.05) in HKPB2 and HKPB3 compared to HKPB1 and the control, no significant difference (P > 0.05) was observed between HKPB2 and HKPB3. The results obtained from the present study indicate that the application of heat-killed L. plantarum at a concentration of 10(8) cfu g(-1) diet, though not effective in augmenting the growth and feed utilization parameters, can significantly improve immune parameters and disease resistance of M. rosenbergii in the laboratory condition. PMID:25542379

  15. Preliminary obervations on use of ethanol distiller's by-products for polyculture of freshwater prawn (Macrobrachium rosenbergii) and golden shiner (Notemigonus crysoleucas)

    SciTech Connect

    Kohler, C.C.; Lewis, W.M.; Krueger, S.P.

    1983-01-01

    Two major by-products of ethanol distillation, corn distiller's solubles and corn distiller's mash, were evaluated for their potential use as a food source for freshwater prawn (Macrobrachium rosenbergii) and golden shiner (Notemigonus crysoleucas) in mid-western United States. Corn distiller's solubles were not found to be highly toxic to aquatic organisms at concentrations ranging up to 10,000 ppM. However, the high biological oxygen demand of the material requires that single applications not exceed 2000 ppM. Yield trials were conducted over a four month period in five 0.1 ha earthen ponds, located in proximity to an ethanol distillery. Each pond was stocked with 2000 postlarval shrimp (0.02 g mean weight). Three of these ponds were also each stocked with 75 adult golden shiner (27.0 g mean weight). One application of corn distiller's solubles (equivalent to approx. 2000 ppM) and three applications of corn distiller's mash (approx. 5 kg/application) were made over the course of the yield trails (approx. 120 days). No other food or fertilizer was added to the ponds. Survival of shrimp ranged from 34 to 75%. Survival of brood fish exceeded 90%. Mean weight of shrimp at harvest was approximately 12 g, and average production was equivalent to approximately 104 kg ha/sup -1/. Golden shiner reared in the same ponds as shrimp had production rates equivalent to approxmately 130 kg ha/sup -1/. The presence of fish did not appear to impede shrimp production. Although production results of fish and shrimp were encouraging, several factors existed which impeded production; these included: (1) little by-product was applied to ponds because the distillery was undergoing its shakedown period; (2) ponds were new and had minimal natural productivity; (3) grass was not fully established on the watershed and ponds became turbid; and (4) lower than normal temperatures prevailed over much of the growing period. 11 references, 2 figures, 8 tables.

  16. Changes of Phosphatidylcholine and Fatty Acids in Germ Cells during Testicular Maturation in Three Developmental Male Morphotypes of Macrobrachium rosenbergii Revealed by Imaging Mass Spectrometry

    PubMed Central

    Siangcham, Tanapan; Chansela, Piyachat; Hayasaka, Takahiro; Masaki, Noritaka; Sroyraya, Morakot; Poljaroen, Jaruwan; Suwansa-ard, Saowaros; Engsusophon, Attakorn; Hanna, Peter J.; Sobhon, Prasert; Setou, Mitsutoshi

    2015-01-01

    Testis maturation, germ cell development and function of sperm, are related to lipid composition. Phosphatidylcholines (PCs) play a key role in the structure and function of testes. As well, increases of polyunsaturated fatty acids (PUFA) and highly unsaturated fatty acids (HUFA), especially arachidonic acid (ARA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) are essential for male fertility. This study is the first report to show the composition and distribution of PCs and total fatty acids (FAs) in three groups of seminiferous tubules (STs) classified by cellular associations [i.e., A (STs with mostly early germ cells), B (STs with mostly spermatids), and C (STs with spermatozoa)], in three morphotypes of Macrobrachium rosenbergii, [i.e., small male (SM), orange claw male (OC), and blue claw male (BC)]. Thin layer chromatography exhibited levels of PCs reaching maxima in STs of group B. Imaging mass spectrometry showed remarkably high signals corresponding to PC (16:0/18:1), PC (18:0/18:2), PC (18:2/20:5), and PC (16:0/22:6) in STs of groups A and B. Moreover, most signals were detected in the early developing cells and the intertubular area, but not at the area containing spermatozoa. Finally, gas chromatography-mass spectrometry indicated that the major FAs present in the testes were composed of 14:0, 16:0, 17:0, 18:0, 16:1, 18:1, 18:2, 20:1, 20:2, 20:4, 20:5, and 22:6. The testes of OC contained the greatest amounts of these FAs while the testes of BC contained the least amounts of these FAs, and there was more EPA (20:5) in the testes of SM and OC than those in the BC. The increasing amounts of FAs in the SM and OC indicate that they are important for spermatogenesis and spermiogenesis. This knowledge will be useful in formulating diets containing PUFA and HUFA for prawn broodstocks in order to improve testis development, and lead to increased male fecundity. PMID:25781176

  17. Dietary supplement of banana (Musa acuminata) peels hot-water extract to enhance the growth, anti-hypothermal stress, immunity and disease resistance of the giant freshwater prawn, Macrobrachium rosenbergii.

    PubMed

    Rattanavichai, Wutti; Cheng, Winton

    2015-04-01

    In the present study, Macrobrachium rosenbergii were fed with diets containing extracts of banana, Musa acuminate, fruit's peel (banana peels extract, BPE) at 0, 1.0, 3.0 and 6.0 g kg(-1). The non-specific immune parameters, disease resistance and anti-hypothermal stress were evaluated at 2, 4, 8, 16 and 32 days of post feeding. Also, we demonstrated the percent weight gain (PWG), percent length gain (PLG), feeding efficiency (FE), and survival rate of giant freshwater prawn at 30, 60, 90, and 120 days of post feeding. The PWG, PLG, FE and survival rate of prawns fed at 0, 1.0, 3.0 and 6.0 g kg(-1) BPE-containing diets after 120 days were 69.5%, 75.4%, 77.8% and 83.3%; 21.8%, 23.6%, 27.8% and 33.9%; 0.60, 0.72, 0.75 and 0.90; and 55.4%, 62.2%, 62.3% and 75.3%, respectively. After 32 days of post feeding, a significant increase in total haemocyte count (THC), different haemocyte count (DHC), respiratory bursts (RBs), superoxide dismutase (SOD) activity, glutathione peroxidase (GPx) activity, phenoloxidase (PO) activity and transglutaminase (TG) activity, and meanwhile, a decreased haemolymph coagulation time was observed. Furthermore, phagocytic activity and clearance efficiency of prawns against Lactococcus garvieae infection were significantly increased. Prawns challenged with L. garvieae after 32 days of feeding at 1.0, 3.0 and 6.0 g kg(-1) had a significantly higher survival rate (33.3%, 40.0% and 56.7%) than those fed with the control diet. Subsequently, hypothermal (14 °C) stress was 43.4%, 50.0% and 50.0%, respectively. Altogether, we therefore recommend the dietary BPE administration at 6.0 g kg(-1) promotes growth, anti-hypothermal stress, and enhance immunity and resistance against L. garvieae in M. rosenbergii. PMID:25634258

  18. The effect of banana (Musa acuminata) peels hot-water extract on the immunity and resistance of giant freshwater prawn, Macrobrachium rosenbergii via dietary administration for a long term: Activity and gene transcription.

    PubMed

    Rattanavichai, Wutti; Chen, Ying-Nan; Chang, Chin-Chyuan; Cheng, Winton

    2015-10-01

    The non-specific immune parameters, disease resistance and immune genes expressions in Macrobrachium rosenbergii were evaluated at 120 days of post feeding the diets containing the extracts of banana, Musa acuminate, fruit's peel (banana peels extract, BPE) at 0, 1.0, 3.0 and 6.0 g kg(-1). Results showed that prawns fed with a diet containing BPE at the level of 1.0, 3.0 and 6.0 g kg(-1) for 120 days had a significantly higher survival rate (30.0%, 40.0% and 56.7%, respectively) than those fed with the control diet after challenge with Lactococcus garvieae for 144 h, and the respective relative survival percentages were 22.2%, 33.3%, and 51.9%, respectively. Dietary BPE supplementation at 3.0 and/or 6.0 g kg(-1) for 120 days showed a significant increase total haemocyte count (THC), granular cell (GC), superoxide dismutase (SOD) activity, phenoloxidase (PO) activity, transglutaminase (TG) activity, and phagocytic activity and clearance efficiency to L. garvieae infection, and meanwhile, the significant decrease in haemolymph clotting times and respiratory bursts (RBs) per haemocyte of prawns were revealed. Furthermore, the mRNA expressions of prophenoloxidase (proPO), lipopolysaccharide and ?-1,3-glucan binding protein (LGBP), peroxinectin (PE), transglutaminase (TG), and crustin (CT) were significantly increased. We therefore recommend that BPE can be used as an immunomodulator for prawns through dietary administration at 6.0 g kg(-1) for a long term (over 120 days) to modify immune responses and genes expression following the enhanced resistance against pathogens. PMID:26118934

  19. Identification of novel allergen in edible insect, Gryllus bimaculatus and its cross-reactivity with Macrobrachium spp. allergens.

    PubMed

    Srinroch, Chutima; Srisomsap, Chantragan; Chokchaichamnankit, Daranee; Punyarit, Phaibul; Phiriyangkul, Pharima

    2015-10-01

    Edible insects have recently been promoted as a source of protein and have a high nutrition value. Identification of allergens and cross-reactivity between Macrobrachium spp. and the field cricket (Gryllus bimaculatus) is necessary for food safety control and to assist in the diagnosis and therapy of allergy symptoms. Denaturing polyacrylamide gel electrophoresis (SDS-PAGE) was used to separate proteins. Allergens were determined and identified by IgE-immunoblotting with pooled sera from prawn-allergic patients (n=16) and LC-MS/MS. Arginine kinase (AK) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were determined as the important allergens in muscle of Macrobrachium rosenbergii whereas, hemocyanin (HC) was identified as an allergen in Macrobrachium spp. The allergens in Macrobrachium lanchesteri were identified as AK and HC. In addition, hexamerin1B (HEX1B) was identified as a novel and specific allergen in G. bimaculatus. The important allergen in G. bimaculatus and Macrobrachium spp. is AK and was found to cross-react between both species. PMID:25872439

  20. Virus-Like Particles of a Fish Nodavirus Display a Capsid Subunit Domain Organization Different from That of Insect Nodaviruses

    E-print Network

    Tang, Liang; Lin, Chan-Shing; Krishna, Neel K.; Yeager, Mark; Schneemann, Anette; Johnson, John E.

    2002-06-01

    single-domain coat protein of insect nodaviruses. The analysis implies that residues 83 to 216 fold as a ?-sandwich which forms the inner shell of the T=3 capsid and residues 217 to 308 form the trimeric surface protrusions observed in the cryoEM map...

  1. Acute toxicity of organochlorine insecticide endosulfan to the giant freshwater prawn Macrobrochium rosenbergii

    NASA Astrophysics Data System (ADS)

    Dai, Xilin; Xiong, Zhaodi; Xie, Jian; Ding, Fujiang

    2014-01-01

    Endosulfan, an organochlorine pesticide, is highly toxic and effective at controlling pests in agriculture, horticulture, and public health programs. In this study, static bioassays were used to evaluate the toxicity of endosulfan to freshwater prawns ( Macrobrachium rosenbergii) of various lengths (1.5±0.03, 4±0.08, and 7±0.06 cm). Additionally, the activities of peroxidase (POD), acid phosphatase (ACP), alkaline phosphatase, acetylcholinesterase (AChE), and Na+/K+-ATPase were analyzed to reflect the effects of endosulfan exposure. The 96 h LC50 of endosulfan for prawns 1.5, 4, and 7 cm long were 1.86, 4.53, and 6.09 ?g/L, respectively, improved tolerance to endosulfan with growth. The POD activities of test organisms exposed to low concentrations of endosulfan were inhibited, indicating the presence of oxygen damaged tissue. Moreover, a notable decrease in AChE activity was observed due to overstimulation of neurotransmission, which might result in abnormal behavior. The effect caused by endosulfan on phosphatase production in the hepatopancreas of prawns 1.5, 4, and 7 cm long was different because the ability of nonspecific immune regulation increased with growth. The 96 h LC50 values obtained in this study could be used in the formulation of water-quality criteria in China. Moreover, the changes in enzymes activities of M. rosenbergii under stress of endosulfan could be applied in the establishment of early warning indicators for bio-safety.

  2. Effect of salt treatments on survival and consumer acceptance of freshwater prawn, Macrobrachium rosenbergii

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Post harvest acclimation of live freshwater prawns to a mixture of water and marine salt increases the consumer acceptability of the finished product. However, the high cost of marine salts prohibits their use in commercial practice. Therefore, the identification of successful, cost effective salt a...

  3. Abundance and diversity of zooplankton in semi intensive prawn (Macrobrachium rosenbergii) farm.

    PubMed

    Shil, Jadobendro; Ghosh, Alokesh Kumar; Rahaman, S M Bazlur

    2013-12-01

    The present study was carried out on the seasonal abundance and diversity of zooplankton in a semi- intensive prawn farm of Bagerhat district from July to December, 2008. Plankton samples were collected by conical shaped monofilament nylon net (Plankton net) and Lugol's solution was used for preservation. The zooplankton abundance was influenced by physico-chemical factors. During the study period 11 genera of zooplankton under 5 orders were recorded from the study ponds namely Copepoda, Rotifera, Cladocera, Ostracoda and Crustacean Larvae. Among all groups copepod was the dominant order. The percentages of Copepoda, Rotifera, Cladocera, Ostracoda and Crustacean Larvae in semi-intensive culture system were 54%, 28%, 12%, 4% and 2% respectively. But the genera Brachionus under the order of Rotifer was dominant among all other genera. Cyclops and Helidiaptomus under the order of Copepod were the 2nd dominant genera. Numbers of zooplankton species were recorded to be the highest in summer season and minimum at early winter season. Highest number of zooplankton found at the month of October. Total zooplankton shows significant positive relationship with water temperature ((r?=?+0.384), Dissolve Oxygen(r?=?+0.113), pH(r?=?+0.320), Free CO2 (r?=?+0.319), Alkalinity(r?=?+0.269), Hardness (r?=?+0.402) and negative relationship with Salinity(r?=?-0.486), Transparency(r?=?-0.693). The findings of the present study will help to improve the management strategies of shrimp culture system. PMID:23667823

  4. An automated microfluidic chip system for detection of piscine nodavirus and characterization of its potential carrier in grouper farms.

    PubMed

    Kuo, Hsiao-Che; Wang, Ting-Yu; Hsu, Hao-Hsuan; Lee, Szu-Hsien; Chen, Young-Mao; Tsai, Tieh-Jung; Ou, Ming-Chang; Ku, Hsiao-Tung; Lee, Gwo-Bin; Chen, Tzong-Yueh

    2012-01-01

    Groupers of the Epinephelus spp. are an important aquaculture species of high economic value in the Asia Pacific region. They are susceptible to piscine nodavirus infection, which results in viral nervous necrosis disease. In this study, a rapid and sensitive automated microfluidic chip system was implemented for the detection of piscine nodavirus; this technology has the advantage of requiring small amounts of sample and has been developed and applied for managing grouper fish farms. Epidemiological investigations revealed an extremely high detection rate of piscine nodavirus (89% of fish samples) from 5 different locations in southern Taiwan. In addition, positive samples from the feces of fish-feeding birds indicated that the birds could be carrying the virus between fish farms. In the present study, we successfully introduced this advanced technology that combines engineering and biological approaches to aquaculture. In the future, we believe that this approach will improve fish farm management and aid in reducing the economic loss experienced by fish farmers due to widespread disease outbreaks. PMID:22912690

  5. An Automated Microfluidic Chip System for Detection of Piscine Nodavirus and Characterization of Its Potential Carrier in Grouper Farms

    PubMed Central

    Hsu, Hao-Hsuan; Lee, Szu-Hsien; Chen, Young-Mao; Tsai, Tieh-Jung; Ou, Ming-Chang; Ku, Hsiao-Tung; Lee, Gwo-Bin; Chen, Tzong-Yueh

    2012-01-01

    Groupers of the Epinephelus spp. are an important aquaculture species of high economic value in the Asia Pacific region. They are susceptible to piscine nodavirus infection, which results in viral nervous necrosis disease. In this study, a rapid and sensitive automated microfluidic chip system was implemented for the detection of piscine nodavirus; this technology has the advantage of requiring small amounts of sample and has been developed and applied for managing grouper fish farms. Epidemiological investigations revealed an extremely high detection rate of piscine nodavirus (89% of fish samples) from 5 different locations in southern Taiwan. In addition, positive samples from the feces of fish-feeding birds indicated that the birds could be carrying the virus between fish farms. In the present study, we successfully introduced this advanced technology that combines engineering and biological approaches to aquaculture. In the future, we believe that this approach will improve fish farm management and aid in reducing the economic loss experienced by fish farmers due to widespread disease outbreaks. PMID:22912690

  6. Crystallization and preliminary crystallographic analysis of a viral RNA-silencing suppressor encoded by Wuhan nodavirus

    PubMed Central

    Liu, Yuanyuan; Zhao, Yuhui; Zhang, Yong; Chen, Hui

    2013-01-01

    Wuhan nodavirus (WhNV), which is a new member of the Nodaviridae family, encodes a viral protein, B2, that suppresses RNA silencing and host-cell RNA interference (RNAi)-mediated immunity. Although Flock House virus (FHV), another member of the Nodaviridae family, also produces a B2 protein with a similar function, the primary sequences of the B2 proteins from WhNV and FHV have no similarity. To gain a better understanding of the structural details and the mechanism of suppression of RNA silencing by WhNV B2 and to compare it with FHV B2, recombinant WhNV B2 protein has been overexpressed in Escherichia coli, purified and crystallized at 291?K using PEG 4000 as a precipitant. A 2.8?Å resolution data set has been collected from a single crystal at 100?K. This crystal belonged to space group P212121, with unit-cell parameters a = 27.3, b = 45.6, c = 133.9?Å, ? = ? = ? = 90°. Assuming the presence of two molecules in the asymmetric unit, the Matthews coefficient is 2.2?Å3?Da?1. PMID:24100569

  7. The Self-Interaction of a Nodavirus Replicase Is Enhanced by Mitochondrial Membrane Lipids

    PubMed Central

    Qiu, Yang; Wang, Zhaowei; Liu, Yongxiang; Han, Yajuan; Miao, Meng; Qi, Nan; Yang, Jie; Xia, Hongjie; Li, Xiaofeng; Qin, Cheng-Feng; Hu, Yuanyang; Zhou, Xi

    2014-01-01

    RNA replication of positive-strand (+)RNA viruses requires the protein-protein interactions among viral replicases and the association of viral replicases with intracellular membranes. Protein A from Wuhan nodavirus (WhNV), which closely associate with mitochondrial membranes, is the sole replicase required for viral RNA replication. Here, we studied the direct effects of mitochondrial membrane lipids (MMLs) on WhNV protein A activity in vitro. Our investigations revealed the self-interaction of WhNV protein A is accomplished via two different patterns (i.e., homotypic and heterotypic self-interactions via different interfaces). MMLs stimulated the protein A self-interaction, and this stimulation exhibited selectivity for specific phospholipids. Moreover, we found that specific phospholipids differently favor the two self-interaction patterns. Furthermore, manipulating specific phospholipid metabolism affected protein A self-interaction and the activity of protein A to replicate RNA in cells. Taken together, our findings reveal the direct effects of membrane lipids on a nodaviral RNA replicase. PMID:24586921

  8. Molecular perspective on the American transisthmian species of Macrobrachium (Caridea, Palaemonidae)

    PubMed Central

    Pileggi, Leonardo G.; Rossi, Natália; Wehrtmann, Ingo S.; Mantelatto, Fernando L.

    2014-01-01

    Abstract The closure of the Isthmus of Panama (about 3.1 million years ago) separated previously continuous populations and created two groups of extant species, which live now in the Pacific and Atlantic drainage systems. This relatively recent event was a trigger to diversification of various species in the Neotropics, nonetheless there are exemplars that do not show sufficient morphologic variability to separate them by traditional morphological tools. About 60 years ago, some freshwater decapod species with high morphological similarity were separate by previous researchers, based on geographical distribution, in Pacific and Atlantic and considered as “sister species”. However, the complete isolation of these prawns by this geographical barrier is questionable, and it has generated doubts about the status of the following transisthmian pairs of sibling species: Macrobrachium occidentale × Macrobrachium heterochirus, Macrobrachium americanum × Macrobrachium carcinus, Macrobrachium digueti × Macrobrachium olfersii, Macrobrachium hancocki × Macrobrachium crenulatum, Macrobrachium tenellum × Macrobrachium acanthurus and Macrobrachium panamense × Macrobrachium amazonicum. Here we evaluated the relation among these pairs of sibling species in a molecular phylogenetic context. We generated 95 new sequences: 26 sequences of 16S rDNA, 25 of COI mtDNA and 44 of 18S nDNA. In total, 181 sequences were analyzed by maximum likelihood phylogenetic method, including 12 Macrobrachium transisthmian species, as well as seven other American Macrobrachium species, and two other palaemonids. Our analysis corroborated the morphological proximity of the sibling species. Despite the high degree of morphological similarities and considerable genetic diversification encountered among the transisthmian sister species, our data support the conclusion that all species included in sibling groups studied herein are valid taxonomic entities, but not all pairs of siblings form natural groups. PMID:25561833

  9. Nodavirus RNA Replication Protein A Induces Membrane Association of Genomic RNA?

    PubMed Central

    Van Wynsberghe, Priscilla M.; Chen, Hau-Ren; Ahlquist, Paul

    2007-01-01

    Positive-strand RNA virus genome replication occurs in membrane-associated RNA replication complexes, whose assembly remains poorly understood. Here we show that prior to RNA replication, the multifunctional, transmembrane RNA replication protein A of the nodavirus flock house virus (FHV) recruits FHV genomic RNA1 to a membrane-associated state in both Drosophila melanogaster and Saccharomyces cerevisiae cells. Protein A has mitochondrial membrane-targeting, self-interaction, RNA-dependent RNA polymerase (RdRp), and RNA capping domains. In the absence of RdRp activity due to an active site mutation (AD692E), protein A stimulated RNA1 accumulation by increasing RNA1 stability. Protein AD692E stimulated RNA1 accumulation in wild-type cells and in xrn1? yeast defective in decapped RNA decay, showing that increased RNA1 stability was not due to protein A-mediated RNA1 recapping. Increased RNA1 stability was closely linked with protein A-induced membrane association of the stabilized RNA and was highly selective for RNA1. Substantial N- and C-proximal regions of protein A were dispensable for these activities. However, increased RNA1 accumulation was eliminated by deleting protein A amino acids (aa) 1 to 370 but was restored completely by adding back the transmembrane domain (aa 1 to 35) and partially by adding back peripheral membrane association sequences in aa 36 to 370. Moreover, although RNA polymerase activity was not required, even small deletions in or around the RdRp domain abolished increased RNA1 accumulation. These and other results show that prior to negative-strand RNA synthesis, multiple domains of mitochondrially targeted protein A cooperate to selectively recruit FHV genomic RNA to membranes where RNA replication complexes form. PMID:17301137

  10. Natural and Experimental Infection of Caenorhabditis Nematodes by Novel Viruses Related to Nodaviruses

    PubMed Central

    Wu, Guang; Nuez, Isabelle; Bélicard, Tony; Jiang, Yanfang; Zhao, Guoyan; Franz, Carl J.; Goldstein, Leonard D.; Sanroman, Mabel; Miska, Eric A.; Wang, David

    2011-01-01

    An ideal model system to study antiviral immunity and host-pathogen co-evolution would combine a genetically tractable small animal with a virus capable of naturally infecting the host organism. The use of C. elegans as a model to define host-viral interactions has been limited by the lack of viruses known to infect nematodes. From wild isolates of C. elegans and C. briggsae with unusual morphological phenotypes in intestinal cells, we identified two novel RNA viruses distantly related to known nodaviruses, one infecting specifically C. elegans (Orsay virus), the other C. briggsae (Santeuil virus). Bleaching of embryos cured infected cultures demonstrating that the viruses are neither stably integrated in the host genome nor transmitted vertically. 0.2 µm filtrates of the infected cultures could infect cured animals. Infected animals continuously maintained viral infection for 6 mo (?50 generations), demonstrating that natural cycles of horizontal virus transmission were faithfully recapitulated in laboratory culture. In addition to infecting the natural C. elegans isolate, Orsay virus readily infected laboratory C. elegans mutants defective in RNAi and yielded higher levels of viral RNA and infection symptoms as compared to infection of the corresponding wild-type N2 strain. These results demonstrated a clear role for RNAi in the defense against this virus. Furthermore, different wild C. elegans isolates displayed differential susceptibility to infection by Orsay virus, thereby affording genetic approaches to defining antiviral loci. This discovery establishes a bona fide viral infection system to explore the natural ecology of nematodes, host-pathogen co-evolution, the evolution of small RNA responses, and innate antiviral mechanisms. PMID:21283608

  11. Establishment of cell lines from a tropical grouper, Epinephelus awoara (Temminck & Schlegel), and their susceptibility to grouper irido- and nodaviruses.

    PubMed

    Lai, Y S; John, J A C; Lin, C H; Guo, I C; Chen, S C; Fang, K; Lin, C H; Chang, C Y

    2003-01-01

    Four tropical marine fish cell lines have been established from the eye, fin, heart and swim bladder of grouper, Epinephelus awoara (Temminck & Schlegel). Optimum media and temperature conditions for maximum growth were standardized. The eye and swim bladder cells were mostly epithelial, but the fin and heart cells were mostly fibroblastic. The viability of cells was 95% after 1 year of storage in liquid nitrogen (-196 degrees C). Besides these four cell lines, previously established grouper brain, kidney and liver cell lines were also used for a viral susceptibility study which showed that all the cell lines were sensitive to grouper iridovirus, whereas only brain, fin and liver cell lines were susceptible to the yellow grouper nervous necrosis virus (a nodavirus). Electron microscopy studies of the grouper irido- and nodaviruses in ultrathin sections of infected cells showed an abundance of viral particles in the cytoplasm of the virus-infected cells indicating the effective replication of these two viruses. It is suggested that these cell lines can be used for the isolation of putative fish specific viruses and provide a valuable tool to study the mechanisms of host-pathogen interactions. Furthermore, these cell lines upon transfection, using pEGFP-C1 and pEGFP-aMT2.5 (ayu metallothionein promoter), produced significant fluorescent signals indicating their utility for exogenous studies. PMID:12962209

  12. A Unique Nodavirus with Novel Features: Mosinovirus Expresses Two Subgenomic RNAs, a Capsid Gene of Unknown Origin, and a Suppressor of the Antiviral RNA Interference Pathway

    PubMed Central

    Schuster, Susan; Zirkel, Florian; Kurth, Andreas; van Cleef, Koen W. R.; Drosten, Christian

    2014-01-01

    ABSTRACT Insects are a reservoir for many known and novel viruses. We discovered an unknown virus, tentatively named mosinovirus (MoNV), in mosquitoes from a tropical rainforest region in Côte d'Ivoire. The MoNV genome consists of two segments of positive-sense RNA of 2,972 nucleotides (nt) (RNA 1) and 1,801 nt (RNA 2). Its putative RNA-dependent RNA polymerase shares 43% amino acid identity with its closest relative, that of the Pariacoto virus (family Nodaviridae). Unexpectedly, for the putative capsid protein, maximal pairwise identity of 16% to Lake Sinai virus 2, an unclassified virus with a nonsegmented RNA genome, was found. Moreover, MoNV virions are nonenveloped and about 50 nm in diameter, larger than any of the known nodaviruses. Mature MoNV virions contain capsid proteins of ?56 kDa, which do not seem to be cleaved from a longer precursor. Northern blot analyses revealed that MoNV expresses two subgenomic RNAs of 580 nt (RNA 3) and 292 nt (RNA 4). RNA 4 encodes a viral suppressor of RNA interference (RNAi) that shares its mechanism with the B2 RNAi suppressor protein of other nodaviruses despite lacking recognizable similarity to these proteins. MoNV B2 binds long double-stranded RNA (dsRNA) and, accordingly, inhibits Dicer-2-mediated processing of dsRNA into small interfering RNAs (siRNAs). Phylogenetic analyses indicate that MoNV is a novel member of the family Nodaviridae that acquired its capsid gene via reassortment from an unknown, distantly related virus beyond the family level. IMPORTANCE The identification of novel viruses provides important information about virus evolution and diversity. Here, we describe an unknown unique nodavirus in mosquitoes, named mosinovirus (MoNV). MoNV was classified as a nodavirus based on its genome organization and on phylogenetic analyses of the RNA-dependent RNA polymerase. Notably, its capsid gene was acquired from an unknown virus with a distant relationship to nodaviruses. Another remarkable feature of MoNV is that, unlike other nodaviruses, it expresses two subgenomic RNAs (sgRNAs). One of the sgRNAs expresses a protein that counteracts antiviral defense of its mosquito host, whereas the function of the other sgRNA remains unknown. Our results show that complete genome segments can be exchanged beyond the species level and suggest that insects harbor a large repertoire of exceptional viruses. PMID:25210176

  13. Effects of hot-water extract of banana (Musa acuminata) fruit's peel on the antibacterial activity, and anti-hypothermal stress, immune responses and disease resistance of the giant freshwater prawn, Macrobrachium rosenbegii.

    PubMed

    Rattanavichai, Wutti; Cheng, Winton

    2014-08-01

    The hot-extracts isolated from fruit's peel of banana, Musa acuminata, was evaluated on the antibacterial activity to pathogens from aquatic animals, and immunostimulating potential, disease resistance and anti-hypothermal stress in giant freshwater prawn, Macrobrachium rosenbergii through injection administration. The banana peel extract (BPE) showed good activity against 1 Gram-positive and 3 Gram-negative pathogens, including Lactococcus garvieae, Photobacteria damsella, Vibrio alginolyticus and Vibrio parahemolyticus especially in prawn pathogen of L. garvieae strain, which were carried out by a disk diffusion method. Prawn received BPE via injection administration at 1-6 ?g (g prawn)(-1) significantly increased total haemocyte count (THC), hyaline cell (HC), granular cell (GC), phenoloxidase (PO) activity and phagocytic activity against L. garvieae from 3 to 6 days, and significantly increased clearance efficiency against L. garvieae and a significantly decreased coagulation time of prawn from 1 to 6 days. Prawn injected with BPE at 6.0 ?g (g prawn)(-1) for 6 days showed significantly increased superoxide dismutase (SOD) activity, but significantly decreased respiratory bursts (RBs) of per haemocyte. Survival rates of M. rosenbergii injected with BPE at concentrations of 1, 3 and 6 ?g (g prawn)(-1) were significantly higher than those injected with saline control after challenge with L. garvieae for 4-6 days, and the respective relative survival percentages of prawn were 28.6%, 38.1%, and 47.8%, respectively at 6 days. The sublethal time of prawns that had received saline and BPE at 1, 3 and 6 ?g (g prawn)(-1) for 6 days and then were transferred from 28 °C to 14 °C were 69.4, 79.8, 83.6, and 90.2 h, respectively. It was concluded that the BPE can be used as the bacteriostat, and immunostimulant and physiological regulator for prawn through injection administration to enhance immunity, physiological responses, and resistance against L. garvieae. PMID:24906123

  14. Functional analysis of an orange-spotted grouper (Epinephelus coioides) interferon gene and characterisation of its expression in response to nodavirus infection.

    PubMed

    Chen, Young-Mao; Kuo, Cham-En; Chen, Guan-Ru; Kao, Yu-Ting; Zou, Jun; Secombes, Chris J; Chen, Tzong-Yueh

    2014-10-01

    We cloned and sequenced 2C I-IFN, a two-cysteine containing type I interferon (I-IFN) gene, in orange-spotted grouper (Epinephelus coioides). The cDNA has 769 base pairs, the protein has 172 amino acids, and the predicted signal peptide has 18 amino acids with two cysteines. This gene is similar to I-FNs from sea bass and other teleosts. 2C I-IFN has 5 exons and 4 introns, also similar to other teleost I-IFNs. Immunohistochemical (IHC) analysis indicated that expression is predominantly membrane-localized in healthy grouper, but has a zonal distribution in nodavirus-infected grouper. Grouper infected with nodavirus had elevated levels of 2C I-IFN at 72 h and Mx at days 6-7. Recombinant 2C I-IFN activated grouper Mx, leading to upregulated antiviral activity. The grouper Mx promoter was highly induced after treatment with recombinant 2C I-IFN. The present results suggest that expression of grouper 2C I-IFN may participate in the immunologic barrier function against nodavirus. PMID:24731841

  15. Bihormonal control of oogenesis in the freshwater prawn, Macrobrachium kistnensis.

    PubMed

    Sarojini, R; Mirajkar, M S; Nagabhushanam, R

    1983-01-01

    The hormonal control of oogenesis has been investigated in the freshwater prawn, Macrobrachium kistnensis. Experiments using bilateral eyestalk ablation and injection of eyestalk extract supported the presence of a gonad inhibiting factor in the eyestalk of female prawns. Injections of brain and thoracic ganglion extract in normal female prawns and in those subjected to bilateral eyestalk ablation revealed the presence of a gonad stimulating factor in the central nervous tissue. PMID:6637521

  16. Regulation of laboratory populations of snails (Biomphalaria and Bulinus spp.) by river prawns, Macrobrachium spp. (Decapoda, Palaemonidae): implications for control of schistosomiasis

    PubMed Central

    Lafferty, Kevin D.; Kuris, Armand M.

    2014-01-01

    Human schistosomiasis is a common parasitic disease endemic in many tropical and subtropical countries. One barrier to achieving long-term control of this disease has been re-infection of treated patients when they swim, bathe, or wade in surface fresh water infested with snails that harbor and release larval parasites. Because some snail species are obligate intermediate hosts of schistosome parasites, removing snails may reduce parasitic larvae in the water, reducing re-infection risk. Here, we evaluate the potential for snail control by predatory freshwater prawns, Macrobrachium rosenbergii and M. vollenhovenii, native to Asia and Africa, respectively. Both prawn species are high value, protein-rich human food commodities, suggesting their cultivation may be beneficial in resource-poor settings where few other disease control options exist. In a series of predation trials in laboratory aquaria, we found both species to be voracious predators of schistosome-susceptible snails, hatchlings, and eggs, even in the presence of alternative food, with sustained average consumption rates of 12% of their body weight per day. Prawns showed a weak preference for Bulinus truncatus over Biomphalaria glabrata snails. Consumption rates were highly predictable based on the ratio of prawn: snail body mass, suggesting satiation-limited predation. Even the smallest prawns tested (0.5–2g) caused snail recruitment failure, despite high snail fecundity. With the World Health Organization turning attention toward schistosomiasis elimination, native prawn cultivation may be a viable snail control strategy that offers a win-win for public health and economic development. PMID:24388955

  17. Genetic variability and phylogenetic aspects in species of the genus Macrobrachium.

    PubMed

    Guerra, A L; Lima, A V B; Lucato Júnior, R V; Chiachio, M C; Taddei, F G; Castiglioni, L

    2014-01-01

    The genus Macrobrachium includes prawns, which are widely distributed in lakes, floodplains, and rivers in tropical and subtropical regions of South America. This genus presents nearly 210 known species with great ecological and economic importance. However, few studies are related to the biology of these crustaceans. In this study, we analyzed the genetic variability and phylogenetic relationship between Macrobrachium amazonicum and Macrobrachium jelskii, which are closely related species. Additionally, they are syntopics and their taxonomy poses problems because it is difficult to differentiate between the species. We used the mitochondrial gene sequences COI and 16S rRNA to assess the genetic structure of these species in 3 populations that were collected from Tiete hydrographic basin (São Paulo State, Brazil). The interspecific comparison of samples that were collected at the same and different locations showed a low rate of genetic variability. This similarity was attributed to the recent introduction of these species in the regions that were sampled and the habitat conditions in which they inhabit. In addition, these results may be consistent with the hypothesis that they are a single species, interspecific hybrids, or metapopulation. The dendrogram analyses did not reveal the formation of clusters, confirming the disturbances in the genetic structure of the samples that were analyzed in this study. These data are pioneers to these crustaceans, and they confirm the ecological and evolutionary problems between these Macrobrachium species. PMID:24854444

  18. Genetic polymorphism, molecular characterization and relatedness of Macrobrachium species (Palaemonidae) based on RAPD-PCR.

    PubMed

    Guerra, A L; Lima, A V B; Taddei, F G; Castiglioni, L

    2010-01-01

    The prawn genus Macrobrachium belongs to the family Palaemonidae. Its species are widely distributed in lakes, reservoirs, floodplains, and rivers in tropical and subtropical regions of South America. Globally, the genus Macrobrachium includes nearly 210 known species, many of which have economic and ecological importance. We analyzed three species of this genus (M. jelskii, M. amazonicum and M. brasiliense) using RAPD-PCR to assess their genetic variability, genetic structure and the phylogenetic relationship between them and to look for molecular markers that enable separation of M. jelskii and M. amazonicum, which are closely related syntopic species. Ten different random decamer primers were used for DNA amplification, yielding 182 fragments. Three of these fragments were monomorphic and exclusive to M. amazonicum or M. jelskii and can be used as specific molecular markers to identify and separate these two species. Similarity indices and a phylogenetic tree showed that M. amazonicum and M. jelskii are closest to each other, while M. brasiliense was the most differentiated species among them; this may be attributed to the different habitat conditions to which these species have been submitted. This information will be useful for further studies on these important crustacean species. PMID:21128212

  19. Vitellogenin and lipovitellin from the prawn Macrobrachium borellii as hydrocarbon pollution biomarker.

    PubMed

    García, C Fernando; Heras, Horacio

    2012-08-01

    During reproduction vitellogenin (VTG) is transported to vitellogenic oocytes as a precursor of egg yolk lipovitellin (LV). As VTG synthesis is affected by environmental stressors, it is widely used as biomarker in endocrine disruption studies. However, it has seldom been employed to evaluate invertebrate hydrocarbon pollution. An ELISA with anti-LV antibody was developed to evaluate the impact of water-soluble fraction of crude oil (WSF) on Macrobrachium borellii vitellogenesis. Prawn VTG concentration was within the range reported for other crustaceans; LV values were positively correlated with gonadosomatic index (GSI). Females at different vitellogenic stages were exposed to a sub-lethal concentration of WSF for 7 days. Exposed animals with GSI>7 increased their VTG and LV titer as compared to control organisms (190% and 140%, respectively). VTG levels in M. borellii were upregulated and highly sensitive to WSF exposure. This assay could be employed as a biomarker for freshwater hydrocarbon pollution. PMID:22704151

  20. 78 FR 50381 - Certain Frozen Warmwater Shrimp From Malaysia: Final Affirmative Countervailing Duty Determination

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-19

    ...merguiensis), fleshy prawn (Penaeus chinensis), giant river prawn (Macrobrachium rosenbergii), giant tiger prawn (Penaeus monodon), redspotted...Program Determined To Be Not Countervailable 1. Human Resource Development Fund C. Programs...

  1. Vacuolating encephalopathy and retinopathy associated with a nodavirus-like agent: a probable cause of mass mortality of wild Golden grey mullet (Liza aurata) and Sharpnose grey mullet (Liza saliens) in Iranian waters of the Caspian Sea.

    PubMed

    Zorriehzahra, Mohammad Jalil; Nazari, Alireza; Ghasemi, Mohaddes; Ghiasi, Maryam; Karsidani, Somayeh Haghighi; Bovo, Giuseppe; Daud, Hassan Hj Mohd

    2014-12-01

    Mullets are dominant fishes in the catch composition in the southern coasts of the Caspian Sea and after (Rutilus frisii kutum Kamensky, 1901) have a worthy role in production of marine proteins and incomings of north provinces of Iran. Mullets stocks decreased dramatically in recent decades in the Caspian Sea and catch amount reached from 6446 MT on 2002 to 2151 MT in 2012. Mysterious mortalities occurred in wild mullet (Liza auratu) and (Liza saliens) in Iranian waters of Caspian Sea in recent years. Regarding to investigation of causative agent of mentioned outbreak about 322 suspected samples were collected from coastal capture sites of Iranian north provinces in 2008 till 2011. Moribund fish revealed skin darkening, erratic swimming, belly-up at rest and high distension of swim bladder. Target tissues such as brain and eye were removed and then fixed for histopathology and TEM assay. Widespread and massive vacuolation were observed in brain, spinal cord, retina and optical nerve and intracytoplasmic vacuoles and virus particles in retina. So concerning to clinical signs, histopathological and TEM findings, it could be concluded that nodavirus-like agent could be probable cause of mass mortality of wild mullet in Iranian waters of the Caspian Sea. PMID:25674618

  2. Toxicity of cryoprotectants agents in freshwater prawn embryos of Macrobrachium amazonicum.

    PubMed

    Ferreira, Arthur Vinícius Lourenço; Castro, Elias José Teles; Barbosa, Mariana Silva Alves; de Sousa, Míriam Luzia Nogueira Martins; de Araújo Neto, Manoel Paiva; Filho, Aldeney Andrade Soares; de Souza Sampaio, Celia Maria

    2015-12-01

    The process of cooling and cryopreservation of prawn embryos is a viable alternative for a continuous supply of larvae for freshwater prawn farming ponds. However, studies involving the application of those techniques as well as on toxicity of cryoprotectants in freshwater prawn embryos are scarce. Thus, this study aims to test the toxicity of methylic alcohol (MET), dimethyl sulfoxide (DMSO) and ethylene glycol (EG) on Macrobrachium amazonicum embryos. For the present experiment, pools of embryos were taken from 15 M. amazonicum females and were divided into three groups and tested in duplicate at concentrations of 10, 5, 3; 1, 0.5 or 0.1%. Toxicity tests were conducted for 24 h in Falcon® pipes to obtain the lethal concentration for 50% of the larvae (LC50). After the set period for testing, random samples of embryos were removed for morphological analysis under stereoscopic microscopes. Results were analysed using analysis of variance (ANOVA) and Tukey's test at a 5% significance level and Trimmed Spearman-Karber Analysis to determine LC50-24 h. DMSO toxicity tests revealed that 5% and 10% concentrations showed the highest toxicity and differed from the control (P ? 0.05), 24h-LC50 was 437.4 ± 14.4 µL. MET was less toxic among the tested cryoprotectants and concentrations did not allow the determination of its LC50-24h. For tests with EG, concentrations of 3, 5 or 10% solutions resulted in a 100% mortality to tested embryos; EG was the tested cryoprotectant with the highest toxicity, with an LC50-24h average of 81.91 ± 35.3 µl. PMID:25255785

  3. Effects of Host Phylogeny and Habitats on Gut Microbiomes of Oriental River Prawn (Macrobrachium nipponense)

    PubMed Central

    Chen, Po-Cheng; Weng, Francis Cheng-Hsuan; Jean, Wen Dar; Wang, Daryi

    2015-01-01

    The gut microbial community is one of the richest and most complex ecosystems on earth, and the intestinal microbes play an important role in host development and health. Next generation sequencing approaches, which rapidly produce millions of short reads that enable the investigation on a culture independent basis, are now popular for exploring microbial community. Currently, the gut microbiome in fresh water shrimp is unexplored. To explore gut microbiomes of the oriental river prawn (Macrobrachium nipponense) and investigate the effects of host genetics and habitats on the microbial composition, 454 pyrosequencing based on the 16S rRNA gene were performed. We collected six groups of samples, including M. nipponense shrimp from two populations, rivers and lakes, and one sister species (M. asperulum) as an out group. We found that Proteobacteria is the major phylum in oriental river prawn, followed by Firmicutes and Actinobacteria. Compositional analysis showed microbial divergence between the two shrimp species is higher than that between the two populations of one shrimp species collected from river and lake. Hierarchical clustering also showed that host genetics had a greater impact on the divergence of gut microbiome than host habitats. This finding was also congruent with the functional prediction from the metagenomic data implying that the two shrimp species still shared the same type of biological functions, reflecting a similar metabolic profile in their gut environments. In conclusion, this study provides the first investigation of the gut microbiome of fresh water shrimp, and supports the hypothesis of host species-specific signatures of bacterial community composition. PMID:26168244

  4. Effects of Host Phylogeny and Habitats on Gut Microbiomes of Oriental River Prawn (Macrobrachium nipponense).

    PubMed

    Tzeng, Tzong-Der; Pao, Yueh-Yang; Chen, Po-Cheng; Weng, Francis Cheng-Hsuan; Jean, Wen Dar; Wang, Daryi

    2015-01-01

    The gut microbial community is one of the richest and most complex ecosystems on earth, and the intestinal microbes play an important role in host development and health. Next generation sequencing approaches, which rapidly produce millions of short reads that enable the investigation on a culture independent basis, are now popular for exploring microbial community. Currently, the gut microbiome in fresh water shrimp is unexplored. To explore gut microbiomes of the oriental river prawn (Macrobrachium nipponense) and investigate the effects of host genetics and habitats on the microbial composition, 454 pyrosequencing based on the 16S rRNA gene were performed. We collected six groups of samples, including M. nipponense shrimp from two populations, rivers and lakes, and one sister species (M. asperulum) as an out group. We found that Proteobacteria is the major phylum in oriental river prawn, followed by Firmicutes and Actinobacteria. Compositional analysis showed microbial divergence between the two shrimp species is higher than that between the two populations of one shrimp species collected from river and lake. Hierarchical clustering also showed that host genetics had a greater impact on the divergence of gut microbiome than host habitats. This finding was also congruent with the functional prediction from the metagenomic data implying that the two shrimp species still shared the same type of biological functions, reflecting a similar metabolic profile in their gut environments. In conclusion, this study provides the first investigation of the gut microbiome of fresh water shrimp, and supports the hypothesis of host species-specific signatures of bacterial community composition. PMID:26168244

  5. Effects of ammonia stress in the Amazon river shrimp Macrobrachium amazonicum (Decapoda, Palaemonidae).

    PubMed

    Pinto, Marcelo R; Lucena, Malson N; Faleiros, Rogério Oliveira; Almeida, Eduardo Alves; McNamara, John C; Leone, Francisco A

    2016-01-01

    We evaluate the effects of total ammonia nitrogen-N (TAN) exposure for 72h on (Na(+),K(+))- and V(H(+))-ATPase activities and on their subunit expressions in gills of the diadromous freshwater shrimp Macrobrachium amazonicum. Specific (Na(+),K(+))- and V(H(+))-ATPase activities increased roughly 1.5- to 2-fold, respectively, after exposure to 2.0mmolL(-1) TAN. Quantitative RT-PCR analyses revealed a 2.5-fold increase in V(H(+))-ATPase B subunit mRNA expression while (Na(+),K(+))-ATPase ?-subunit expression was unchanged. Immunohistochemical analyses of the gill lamellae located the (Na(+),K(+))-ATPase throughout the intralamellar septal cells, independently of TAN concentration, while the V(H(+))-ATPase was located in both the apical pillar cell flanges and pillar cell bodies. Systemic stress parameters like total hemocyte count decreased by 30% after exposure to 2.0mmolL(-1) TAN, accompanied by increased activities of the oxidative stress enzymes superoxide dismutase, glutathione reductase and glucose-6-phosphate dehydrogenase in the gills. The stress responses of M. amazonicum to elevated TAN include increases in gill (Na(+),K(+))- and V(H(+))-ATPase activities that are accompanied by changes in oxidative stress enzyme activities, immune system effects and an increase in gill V(H(+))-ATPase gene expression. These findings likely underpin physiological effects in a crustacean like M. amazonicum that exploits multiple ecosystems during its life cycle, as well as under culture conditions that may significantly impact shrimp production by the aquaculture industry. PMID:26571214

  6. cDNA Cloning and Expression Analysis of Gustavus Gene in the Oriental River Prawn Macrobrachium nipponense

    PubMed Central

    Zhang, Fengying; Chen, Liqiao; Qin, Jianguang; Zhao, Weihong; Wu, Ping; Yu, Na; Ma, Lingbo

    2011-01-01

    The gustavus gene is required for localizing pole plasm and specifying germ cells. Research on gustavus gene expression will advance our understanding of the biological function of gustavus in animals. A cDNA encoding gustavus protein was identified and termed MnGus in the oriental river prawn Macrobrachium nipponense. Bioinformatic analyses showed that this gene encoded a protein of 262 amino acids and the protein belongs to the Spsb1 family. Real-time quantitative PCR analyses revealed that the expression level of MnGus in prawn embryos was slightly higher at the cleavage stage than at the blastula stage, and reached the maximum level during the zoea stage of embryos. The minimum level of MnGus expression occurred during the perinucleolus stage in the ovary, while the maximum was at the oil globule stage, and then the level of MnGus expression gradually decreased with the advancement of ovarian development. The expression level of MnGus in muscle was much higher than that in other tissues in mature prawn. The gustavus cDNA sequence was firstly cloned from the oriental river prawn and the pattern of gene expression was described during oocyte maturation, embryonic development, and in other tissues. The differential expression patterns of MnGus in the embryo, ovary and other somatic tissues suggest that the gustavus gene performs multiple physiological functions in the oriental river prawn. PMID:21359189

  7. Molecular Analysis of the Freshwater Prawn Macrobrachium olfersii (Decapoda, Palaemonidae) Supports the Existence of a Single Species throughout Its Distribution

    PubMed Central

    Rossi, Natália; Mantelatto, Fernando Luis

    2013-01-01

    Macrobrachium olfersii is an amphidromous freshwater prawn, widespread along the eastern coasts of the Americas. This species shows great morphological modifications during ontogenesis, and several studies have verified the existence of a wide intraspecific variation. Because of this condition, the species is often misidentified, and several synonyms have been documented. To elucidate these aspects, individuals of M. olfersii from different populations along its range of distribution were investigated. The taxonomic limit was established, and the degree of genetic variability of this species was described. We extracted DNA from 53 specimens of M. olfersii, M. americanum, M. digueti and M. faustinum, which resulted in 84 new sequences (22 of 16S mtDNA, 45 of Cythocrome Oxidase I (COI) mtDNA, and 17 of Histone (H3) nDNA). Sequences of three genes (single and concatenated) from these species were used in the Maximum Likelihood and Bayesian Inference phylogenetic analyses and COI sequences from M. olfersii were used in population analysis. The genetic variation was evaluated through the alignment of 554 bp from the 16S, 638 bp from the COI, and 338 bp from the H3. The rates of genetic divergence among populations were lower at the intraspecific level. This was confirmed by the haplotype net, which showed a continuous gene flow among populations. Although a wide distribution and high morphological intraspecific variation often suggest the existence of more than one species, genetic similarity of Caribbean and Brazilian populations of M. olfersii supported them as a single species. PMID:23382941

  8. Molecular characterization of a novel ovary-specific gene fem-1 homolog from the oriental river prawn, Macrobrachium nipponense.

    PubMed

    Ma, Ke-Yi; Liu, Zhi-Qiang; Lin, Jing-Yun; Li, Jia-Le; Qiu, Gao-Feng

    2016-01-10

    The feminization-1 (fem-1) gene is characterized by one of the most common protein-protein interaction motifs, ankyrin repeat motifs, displays many expression patterns in vertebrates and invertebrates, and plays an essential role in the sex-determination/differentiation pathway in Caenorhabditis elegans. In this study, a fem-1 homolog, designated as Mnfem-1, was first cloned from the oriental river prawn Macrobrachium nipponense. The prawn Mnfem-1 gene consists of six exons and five introns. The full-length cDNA (2603bp) of Mnfem-1 contains an open reading frame (ORF) encoding a protein of 622 amino acids. The Mnfem-1 RNA and protein are exclusively expressed in the ovary in adult prawns as revealed by RT-PCR and immunofluorescence analysis, respectively. In situ hybridization results showed that strong positive signals were concentrated at the edge of the previtellogenic and vitellogenic oocyte. During embryogenesis, Mnfem-1 is highly expressed in both unfertilized eggs and embryos at cleavage stage and thereafter dropped to a low level from blastula to zoea, indicating that the Mnfem-1 in early embryos is maternal. After hatching, the Mnfem-1 expression significantly increased in the larvae at length of 2cm, an important stage of sex differentiation. Yeast two hybridization results showed that the Mnfem-1 protein can be potentially interactive with cathepsin L and proteins containing the domains of insulinase, ankyrin or ubiquitin. Our results suggested that Mnfem-1 could have roles in prawn ovarian development and sex determination/differentiation. PMID:26367327

  9. Ocurrence of male morphotypes of Macrobrachium amazonicum (Caridea, Palaemonidae) in a population with an entirely freshwater life cycle.

    PubMed

    Pantaleão, J A F; Hirose, G L; Costa, R C

    2014-08-01

    This study records, for the first time, the occurrence of all four male morphotypes in a population of Macrobrachium amazonicum from a continental environment, with an entirely freshwater life cycle. The specimens studied came from the Tietê River, state of São Paulo, Brazil, and were collected in a lotic environment downstream from Ibitinga Dam. This population was compared with other continental populations, including a population from the dam itself, collected in a previous study. Four samples of 30 minutes were taken monthly, using a trap, from January to April 2011. Each male specimen was measured with respect to seven body dimensions as follows: carapace length (CL), right cheliped length (RCL), dactyl length (DCL), propodus length (PPL), carpus length (CRL), merus length (ML) and ischium length (IL). The relative growth was analyzed based on the change in growth patterns of certain body parts in relation to the independent variable CL. The four male morphotypes proposed for the species were found using morphological and morphometric analyses. Different biological characteristics were found between the populations studied. The male population of the lake of Ibitinga and from Pantanal presented mean sizes and number of morphotypes lower than the population studied here. These differences seem to be closely related to ecological characteristics of the environments inhabited by these populations. Our results supported the hypothesis that coastal and continental populations of M. amazonicum belong to the same species. PMID:25627389

  10. Larval Pseudoproleptus sp. (Nematoda: Cystidicolidae) found in the Amazon river prawn Macrobrachium amazonicum (Decapoda: Palaemonidae) in Brazil.

    PubMed

    Moravec, Frantisek; Santos, Cláudia P

    2009-06-01

    Previously undescribed infective larvae of the cystidicolid nematode Pseudoproleptus sp. (probably conspecific with the nematode originally described as Heliconema izecksohni Fabio, 1982, a parasite of freshwater fish in Brazil), were found encapsulated in the hemocel of the Amazon river prawn Macrobrachium amazonicum (Heller) (Decapoda: Palaemonidae) from the natural canals on the Mexiana Island (Amazon River Delta), Pardá State, Brazil. The prevalence in prawns (body length 48-110 mm) examined in January and March 2008 (n = 44) was 32%, with an intensity of 1-6 (mean 2) larvae per crustacean. The nematode larvae (body length 19.7-25.7 mm), characterized by the cephalic end provided with a helmet-like cuticular structure having a thickened free posterior margin, are described based on light and scanning electron microscopy. Apparently prawns play a role as intermediate hosts for this nematode species. This is the first record of a larval representative of Cystidicolidae in South America and the first record of a species of Pseudoproleptus Khera, 1955, in the Neotropics. Heliconema izecksohni is transferred to Pseudoproleptus as Pseudoproleptus izecksohni (Fabio, 1982) n. comb. PMID:19014207

  11. Isolation and characterization of two novel C-type lectins from the oriental river prawn, Macrobrachium nipponense.

    PubMed

    Xiu, Yunji; Hou, Libo; Liu, Xiaoqian; Wang, Yinghui; Gu, Wei; Meng, Qingguo; Wang, Wen

    2015-10-01

    C-type lectins are a family of calcium-dependent carbohydrate-binding proteins which are believed to play important roles in the innate immunity of invertebrates. This study identified two novel C-type lectins, designated as MnCTLDcp2 and MnCTLDcp3, from the oriental river prawn Macrobrachium nipponense. The full-length cDNA of MnCTLDcp2 was of 1582 bp with an open reading frame (ORF) of 972 bp encoding a polypeptide of 323 amino acids. The complete nucleotide sequence of MnCTLDcp3 cDNA was 583 bp, containing a 555 bp ORF encoding a putative protein of 184 deduced amino acids. The deduced MnCTLDcp2 and MnCTLDcp3 proteins both contained a single C-type lectin-like domain (CTLD). Besides, MnCTLDcp2 contains a signal peptide and an low-density lipoprotein receptor class A (LDLa) domain. Reverse transcription PCR showed that MnCTLDcp2 was expressed in the heart, gill, nerve hepatopancreas and intestine; MnCTLDcp3 was expressed in the hepatopancreas, heart, nerve, gill and muscle. Their expression in the heart tissue was regulated following challenge with bacteria. The microbial agglutination assay showed that both MnCTLDcp2 and MnCTLDcp3 could agglutinated bacteria in the presence of calcium. All these results suggested that MnCTLDcp2 and MnCTLDcp3 functioned as pattern recognition receptors in the immune system of M. nipponense. PMID:26208755

  12. Transcriptome Analysis of the Oriental River Prawn, Macrobrachium nipponense Using 454 Pyrosequencing for Discovery of Genes and Markers

    PubMed Central

    Ma, Keyi; Qiu, Gaofeng; Feng, Jianbin; Li, Jiale

    2012-01-01

    Background The oriental river prawn, Macrobrachium nipponense, is an economically and nutritionally important species of the Palaemonidae family of decapod crustaceans. To date, the sequencing of its whole genome is unavailable as a non-model organism. Transcriptomic information is also scarce for this species. In this study, we performed de novo transcriptome sequencing to produce the first comprehensive expressed sequence tag (EST) dataset for M. nipponense using high-throughput sequencing technologies. Methodology and Principal Findings Total RNA was isolated from eyestalk, gill, heart, ovary, testis, hepatopancreas, muscle, and embryos at the cleavage, gastrula, nauplius and zoea stages. Equal quantities of RNA from each tissue and stage were pooled to construct a cDNA library. Using 454 pyrosequencing technology, we generated a total of 984,204 high quality reads (338.59Mb) with an average length of 344 bp. Clustering and assembly of these reads produced a non-redundant set of 81,411 unique sequences, comprising 42,551 contigs and 38,860 singletons. All of the unique sequences were involved in the molecular function (30,425), cellular component (44,112) and biological process (67,679) categories by GO analysis. Potential genes and their functions were predicted by KEGG pathway mapping and COG analysis. Based on our sequence analysis and published literature, many putative genes involved in sex determination, including DMRT1, FTZ-F1, FOXL2, FEM1 and other potentially important candidate genes, were identified for the first time in this prawn. Furthermore, 6,689 SSRs and 18,107 high-confidence SNPs were identified in this EST dataset. Conclusions The transcriptome provides an invaluable new data for a functional genomics resource and future biological research in M. nipponense. The molecular markers identified in this study will provide a material basis for future genetic linkage and quantitative trait loci analyses, and will be essential for accelerating aquaculture breeding programs with this species. PMID:22745820

  13. Manifestation of carbaryl toxicity on soluble protein and histopathology in the hepatopancreas and gills of the prawn, Macrobrachium malcolmsonii.

    PubMed

    Bhavan, P Saravana; Geraldine, P

    2009-07-01

    The present study examined alterations in the protein pattern and histopathology of the hepatopancreas and gills of Macrobrachium malcolmsonii following exposure to Sevin, the commercial grade of carbaryl (1-Naphthylmethyl carbamate) pesticide. Juvenile prawns were exposed to sub-lethal concentrations 5.15 microg l(-1), 7.73 microg l(-1) and 15.47 microg l(-1) of carbaryl for 21 days. Samples were obtained from the hepatopancreas and gills of prawns. The concentration of buffer soluble protein in tissues of test prawns was significantly (p<2.5%) lower than the control. This suggests that proteins were utilized to generate energy in order to withstand carbaryl induced toxic stress. Polypeptide bands of different molecular mass (150-10 kDa) in tissues of test prawns stained less intensely than those in control. This indicates that carbaryl toxicity caused protein degradation in tissues of test prawns. The histology of the hepatopancreas and gills showed few marked pathological changes in prawns exposed to carbaryl. These include rupture of the basal laminae, abnormal infiltration/aggregation of hemocytes in the interstitial sinus of the hepatopancreas and in the hemocoelic space of gill lamellae, fused gill lamellae and necrosis like appearance both in the tubules of the hepatopancreas and gill lamellae. The structural alterations observed in the hepatopancreas and gills of the prawns suggest that carbaryl caused tissue damage in M. malcolmsonii. Protein degradation is a phenomenon in damaging tissues, and thus, under such condition the hepatopancreas and gills of test prawns would not perform their vital functions, which in turn ultimately affect the survival and growth of M. malcolmsonii. The changes noted in protein and histology of test prawns can be taken as biomarkers for monitoring pesticide pollution in aquatic environments. Since this species of prawn is a nutritious delicacy to human being, monitoring aquatic pollution by carbaryl is warranted. PMID:20120492

  14. Toxicity of the organophosphorous insecticide metamidophos (o,s-dimethyl phosphoramidothioate) to larvae of the freshwater prawn and the blue shrimp

    SciTech Connect

    Juarez, L.M.; Sanchez, J. )

    1989-08-01

    The organophosphorous insecticide O,S-dimethyl phosphoramidothioate (Metamidophos, Tamaron, Monitor, Hamidop) is widely used for pest control in tropical crops. If washed down to streams and estuaries its residues could adversely affect populations of commercially important crustaceans, like those of the palaemonid prawn Macrobrachium rosenbergii and the penaeid shrimp Penaeus stylirostris. This paper presents information on the toxicity of O,S-dimethyl phosphoramidothioate to larvae of M. rosenbergii and P. stylirostris.

  15. The effect of astaxanthin on resistance of juvenile prawns Macrobrachium nipponense (Decapoda: Palaemonidae) to physical and chemical stress.

    PubMed

    Tizkar, Babak; Seidavi, Alireza; Ponce-Palafox, Jesús Trinidad; Pourashoori, Parastoo

    2014-12-01

    In recent years, the use of new scientific techniques has effectively improved aquaculture production processes. Astaxanthin has various properties in aquaculture and its antioxidant benefits have been closely related to stress resistance; besides, it is an essential factor for growth in many crustaceans and fish. The objective of this study was to evaluate the resistance of prawn (Macrobrachium nipponense) fed diets containing different amounts of astaxanthin (AX) to the shock and stress of different physicochemical environments. A 70-day trial was conducted to evaluate the effect of supplementation of a source of astaxanthin (Carophyll Pink, 10% astaxanthin, w/w, Hoffman-La Roche, Switzerland) at various levels in the diet of M. nipponense juveniles. Four dry diets were prepared: AX0 without astaxanthin, AX50 with 50 mg/kg, AX100 with 100 mg/kg, and AX150 with 150 mg/kg astaxanthin. The feeding trial was conducted in a recirculation water system consisting of 12 fiberglass tanks (1000L) used for holding prawns. Three replicate aquaria were initially stocked with 36 org/m2 per tank. During the trial, prawns were maintained on a 12:12-h light:dark photoperiod with an ordinary incandescent lamp, and the water quality parameters were maintained as follows: water temperature, 25-26°C; salinity, 1 g/L; pH, 8.5-8.8; dissolved oxygen, 6.0-6.5 mg/L; and ammonia-nitrogen, 0.05 mg/L. Incorporation of AX, production output, and physiological condition were recorded after 10 weeks of feeding. At the end of the growing period, the prawns were exposed to thermal shock (0°C), ammonia (0.75 mg/L), and reduced oxygen (0.5 mg/L). The time to lethargy and the time to complete death of the prawns were recorded. The results showed that control prawns had the shortest time to lethargy and death compared with prawns subjected to the other treatments. The results of this study have shown that the amount of muscle tissue and gill carotenoids in prawn fed with an AX150 diet showed greater reduction than those exposed to other treatments. It is possible that higher levels of astaxanthin in the body under oxygen reduction stress can be beneficial for prawns. These results suggest that male prawns showed lethargy earlier than females, and the percentage of carotenoid reduction in muscle and gill tissues was higher in males. PMID:25720170

  16. The prawn Macrobrachium vollenhovenii in the Senegal River basin: towards sustainable restocking of all-male populations for biological control of schistosomiasis.

    PubMed

    Savaya Alkalay, Amit; Rosen, Ohad; Sokolow, Susanne H; Faye, Yacinthe P W; Faye, Djibril S; Aflalo, Eliahu D; Jouanard, Nicolas; Zilberg, Dina; Huttinger, Elizabeth; Sagi, Amir

    2014-08-01

    Early malacological literature suggests that the outbreak of schistosomiasis, a parasitic disease transmitted by aquatic snails, in the Senegal River basin occurred due to ecological changes resulting from the construction of the Diama dam. The common treatment, the drug praziquantel, does not protect from the high risk of re-infection due to human contact with infested water on a daily basis. The construction of the dam interfered with the life cycle of the prawn Macrobrachium vollenhovenii by blocking its access to breeding grounds in the estuary. These prawns were demonstrated to be potential biological control agents, being effective predators of Schistosoma-susceptible snails. Here, we propose a responsible restocking strategy using all-male prawn populations which could provide sustainable disease control. Male prawns reach a larger size and have a lower tendency to migrate than females. We, therefore, expect that periodic restocking of all-male juveniles will decrease the prevalence of schistosomiasis and increase villagers' welfare. In this interdisciplinary study, we examined current prawn abundance along the river basin, complemented with a retrospective questionnaire completed by local fishermen. We revealed the current absence of prawns upriver and thus demonstrated the need for restocking. Since male prawns are suggested to be preferable for bio-control, we laid the molecular foundation for production of all-male M. vollenhovenii through a complete sequencing of the insulin-like androgenic gland-encoding gene (IAG), which is responsible for sexual differentiation in crustaceans. We also conducted bioinformatics and immunohistochemistry analyses to demonstrate the similarity of this sequence to the IAG of another Macrobrachium species in which neo-females are produced and their progeny are 100% males. At least 100 million people at risk of schistosomiasis are residents of areas that experienced water management manipulations. Our suggested non-breeding sustainable model of control-if proven successful-could prevent re-infections and thus prove useful throughout the world. PMID:25166746

  17. The Prawn Macrobrachium vollenhovenii in the Senegal River Basin: Towards Sustainable Restocking of All-Male Populations for Biological Control of Schistosomiasis

    PubMed Central

    Savaya Alkalay, Amit; Rosen, Ohad; Sokolow, Susanne H.; Faye, Yacinthe P. W.; Faye, Djibril S.; Aflalo, Eliahu D.; Jouanard, Nicolas; Zilberg, Dina; Huttinger, Elizabeth; Sagi, Amir

    2014-01-01

    Early malacological literature suggests that the outbreak of schistosomiasis, a parasitic disease transmitted by aquatic snails, in the Senegal River basin occurred due to ecological changes resulting from the construction of the Diama dam. The common treatment, the drug praziquantel, does not protect from the high risk of re-infection due to human contact with infested water on a daily basis. The construction of the dam interfered with the life cycle of the prawn Macrobrachium vollenhovenii by blocking its access to breeding grounds in the estuary. These prawns were demonstrated to be potential biological control agents, being effective predators of Schistosoma-susceptible snails. Here, we propose a responsible restocking strategy using all-male prawn populations which could provide sustainable disease control. Male prawns reach a larger size and have a lower tendency to migrate than females. We, therefore, expect that periodic restocking of all-male juveniles will decrease the prevalence of schistosomiasis and increase villagers' welfare. In this interdisciplinary study, we examined current prawn abundance along the river basin, complemented with a retrospective questionnaire completed by local fishermen. We revealed the current absence of prawns upriver and thus demonstrated the need for restocking. Since male prawns are suggested to be preferable for bio-control, we laid the molecular foundation for production of all-male M. vollenhovenii through a complete sequencing of the insulin-like androgenic gland-encoding gene (IAG), which is responsible for sexual differentiation in crustaceans. We also conducted bioinformatics and immunohistochemistry analyses to demonstrate the similarity of this sequence to the IAG of another Macrobrachium species in which neo-females are produced and their progeny are 100% males. At least 100 million people at risk of schistosomiasis are residents of areas that experienced water management manipulations. Our suggested non-breeding sustainable model of control—if proven successful—could prevent re-infections and thus prove useful throughout the world. PMID:25166746

  18. A Simple "in Vitro" Culture of Freshwater Prawn Embryos for Laboratory Investigations

    ERIC Educational Resources Information Center

    Porntrai, Supaporn; Damrongphol, Praneet

    2008-01-01

    Giant freshwater prawn ("Macrobrachium rosenbergii" De Man) embryos can be cultured "in vitro" to hatching in 15% (v/v) artificial seawater (ASW). This technique can be applied as a bioassay for testing toxicity or for the effects of various substances on embryo development and can be used as a simple and low-cost model for studying embryo…

  19. Marine Fisheries On the cover

    E-print Network

    Marine Fisheries ~~WD~~ On the cover: A spring chinook gillnetter on the Columbia River at Astoria, Macrobrachium rosenbergii Departments NOAA/NMFS Developments Foreign Fishery Developments Index Papers in Marine For Fisheries National Marine Fisheries Service Editor: W. Hobart Marine Fisheries Review (USPS 090-080) is pub

  20. Transcriptome Analysis of Androgenic Gland for Discovery of Novel Genes from the Oriental River Prawn, Macrobrachium nipponense, Using Illumina Hiseq 2000

    PubMed Central

    Jin, Shubo; Fu, Hongtuo; Zhou, Qiao; Sun, Shengming; Jiang, Sufei; Xiong, Yiwei; Gong, Yongsheng; Qiao, Hui; Zhang, Wenyi

    2013-01-01

    Background The oriental river prawn, Macrobrachium nipponense, is an important aquaculture species in China, even in whole of Asia. The androgenic gland produces hormones that play crucial roles in sexual differentiation to maleness. This study is the first de novo M. nipponense transcriptome analysis using cDNA prepared from mRNA isolated from the androgenic gland. Illumina/Solexa was used for sequencing. Methodology and Principal Finding The total volume of RNA sample was more than 5 ug. We generated 70,853,361 high quality reads after eliminating adapter sequences and filtering out low-quality reads. A total of 78,408 isosequences were obtained by clustering and assembly of the clean reads, producing 57,619 non-redundant transcripts with an average length of 1244.19 bp. In total 70,702 isosequences were matched to the Nr database, additional analyses were performed by GO (33,203), KEGG (17,868), and COG analyses (13,817), identifying the potential genes and their functions. A total of 47 sex-determination related gene families were identified from the M. nipponense androgenic gland transcriptome based on the functional annotation of non-redundant transcripts and comparisons with the published literature. Furthermore, a total of 40 candidate novel genes were found, that may contribute to sex-determination based on their extremely high expression levels in the androgenic compared to other sex glands,. Further, 437 SSRs and 65,535 high-confidence SNPs were identified in this EST dataset from which 14 EST-SSR markers have been isolated. Conclusion Our study provides new sequence information for M. nipponense, which will be the basis for further genetic studies on decapods crustaceans. More importantly, this study dramatically improves understanding of sex-determination mechanisms, and advances sex-determination research in all crustacean species. The huge number of potential SSR and SNP markers isolated from the transcriptome may shed the lights on research in many fields, including the evolution and molecular ecology of Macrobrachium species. PMID:24204682

  1. cDNA cloning and expression of Ubc9 in the developing embryo and ovary of Oriental river prawn, Macrobrachium nipponense.

    PubMed

    Zhang, Fengying; Chen, Liqiao; Wu, Ping; Zhao, Weihong; Li, Erchao; Qin, Jianguang

    2010-03-01

    The small ubiquitin-like modifier (SUMO) pathway in eukaryotes is an essential biological process involving cellular processes, development and organelle biogenesis. In a sequential enzymatic action, Ubc9 is an important conjunction enzyme in the SUMO pathway. Although the Ubc9 has been found in vertebrates, its expression in crustaceans is little known. In this study, the Ubc9 was identified in the embryo and ovary of a freshwater prawn Macrobrachium nipponense for the first time and it was denoted as MnUbc9. Bioinformatics analyses showed that this gene encodes a protein of 161 amino acids with predicted molecular mass of 18.32kDa. Real-time quantitative PCR analyses demonstrated that the expression levels varied significantly in the developing embryo and ovary. In the embryo, the expression level of MnUbc9 was higher at the cleavage stage (CS) than at the blastula stage (BS), and reached even higher levels at the protozoea stage (PS) and the zoea stage (ZS). In the ovary, the MuUbc9 expression was low at the early stage, but reached the highest at the yolk granule stage (YG), and then abruptly declined at the maturation stage (MA). The differential expressions of MnUbc9 in the embryo and ovary suggest that MnUbc9 may play an important role in embryogenesis and oogenesis of M. nipponense. PMID:19944179

  2. Molecular characterization of cathepsin L cDNA and its expression during oogenesis and embryogenesis in the oriental river prawn Macrobrachium nipponense (Palaemonidae).

    PubMed

    Zhao, W; Chen, L; Zhang, F; Wu, P; Li, E; Qin, J

    2013-01-01

    We identified the cDNA sequence of cathepsin L (CatL) in Macrobrachium nipponense, designated as MnCatL, for the first time. The MnCatL cDNA, isolated from the ovary, was 1710 bp in length, containing a 31-bp 5'-untranslated region, a 650-bp 3'-untranslated region, and an open reading frame of 1029 bp, encoding a 342-amino acid polypeptide with a predicted molecular mass of 37.7 kDa. The polypeptide is composed of an 18-amino acid signal peptide, a 106-amino acid propeptide and a 218-amino acid mature peptide. MnCatL mRNA was detected in all tissues that we examined, including the thoracic ganglia, heart, muscle, intestine, hemocytes, ovary, testis, gills, and hepatopancreas. MnCatL expression reached a maximum value in both hepatopancreas and ovaries at the later stages of vitellogenesis, suggesting that MnCatL is involved in ovarian maturation of the oriental river prawn. During embryogenesis, MnCatL expression decreased as the embryo developed. The expression of MnCatL in the ovary and embryo suggest that MnCatL plays an important role in the uptake of vitellogenin and yolk protein, which are deposited in the oocyte for ovary maturation and embryo development, during oogenesis and embryogenesis of M. nipponense. PMID:24301782

  3. Activity and Transcriptional Responses of Hepatopancreatic Biotransformation and Antioxidant Enzymes in the Oriental River Prawn Macrobrachium nipponense Exposed to Microcystin-LR.

    PubMed

    Yuan, Julin; Wang, Xueqin; Gu, Zhiming; Zhang, Yingying; Wang, Zaizhao

    2015-01-01

    Microcystins (MCs) are a major group of cyanotoxins with side effects in many organisms; thus, compounds in this group are recognized as potent stressors and health hazards in aquatic ecosystems. In order to assess the toxicity of MCs and detoxification mechanism of freshwater shrimp Macrobrachium nipponense, the full-length cDNAs of the glutathione S-transferase (gst) and catalase (cat) genes were isolated from the hepatopancreas. The transcription level and activity changes in the biotransformation enzyme (glutathione S-transferase (GST)) and antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx)) in the hepatopancreas of M. nipponense exposed to MC-LR (0.2, 1, 5, and 25 ?g/L) for 12, 24, 72 and 96 h were analyzed. The results showed that the isolated full-length cDNAs of cat and gst genes from M. nipponense displayed a high similarity to other crustaceans, and their mRNAs were mainly expressed in the hepatopancreas. MC-LR caused significant increase of GST activity following 48-96 h (p < 0.05) and an increase in SOD activity especially in 24- and 48-h exposures. CAT activity was activated when exposed to MC-LR in 12-, 24- and 48-h exposures and then it was inhibited at 96-h exposure. There was no significant effect on GPx activity after the 12- and 24-h exposures, whereas it was significantly stimulated after the 72- and 96-h exposures (p < 0.05). The transcription was altered similarly to enzyme activity, but the transcriptional response was generally more immediate and had greater amplitude than enzymatic response, particularly for GST. All of the results suggested that MC-LR can induce antioxidative modulation variations in M. nipponense hepatopancreas in order to eliminate oxidative damage. PMID:26457718

  4. Activity and Transcriptional Responses of Hepatopancreatic Biotransformation and Antioxidant Enzymes in the Oriental River Prawn Macrobrachium nipponense Exposed to Microcystin-LR

    PubMed Central

    Yuan, Julin; Wang, Xueqin; Gu, Zhiming; Zhang, Yingying; Wang, Zaizhao

    2015-01-01

    Microcystins (MCs) are a major group of cyanotoxins with side effects in many organisms; thus, compounds in this group are recognized as potent stressors and health hazards in aquatic ecosystems. In order to assess the toxicity of MCs and detoxification mechanism of freshwater shrimp Macrobrachium nipponense, the full-length cDNAs of the glutathione S-transferase (gst) and catalase (cat) genes were isolated from the hepatopancreas. The transcription level and activity changes in the biotransformation enzyme (glutathione S-transferase (GST)) and antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx)) in the hepatopancreas of M. nipponense exposed to MC-LR (0.2, 1, 5, and 25 ?g/L) for 12, 24, 72 and 96 h were analyzed. The results showed that the isolated full-length cDNAs of cat and gst genes from M. nipponense displayed a high similarity to other crustaceans, and their mRNAs were mainly expressed in the hepatopancreas. MC-LR caused significant increase of GST activity following 48–96 h (p < 0.05) and an increase in SOD activity especially in 24- and 48-h exposures. CAT activity was activated when exposed to MC-LR in 12-, 24- and 48-h exposures and then it was inhibited at 96-h exposure. There was no significant effect on GPx activity after the 12- and 24-h exposures, whereas it was significantly stimulated after the 72- and 96-h exposures (p < 0.05). The transcription was altered similarly to enzyme activity, but the transcriptional response was generally more immediate and had greater amplitude than enzymatic response, particularly for GST. All of the results suggested that MC-LR can induce antioxidative modulation variations in M. nipponense hepatopancreas in order to eliminate oxidative damage. PMID:26457718

  5. Direct use of low temperature geothermal water by Aquafarms International, Inc. for freshwater aquaculture (prawns and associated species). An operations and maintenance manual

    SciTech Connect

    Broughton, R.; Price, M.; Price, V.; Grajcer, D.

    1984-04-01

    In connection with an ongoing commercial aquaculture project in the Coachella Valley, California; a twelve month prawn growout demonstration project was conducted. This project began in August, 1979 and involved the use of low temperature (85/sup 0/F) geothermal waters to raise freshwater prawns, Macrobrachium rosenbergii (deMan), in earthen ponds. The following publication is an operations and maintenance guide which may by useful for those interested in conducting similar enterprises.

  6. Geothermal aquaculture: a guide to freshwater prawn culture

    SciTech Connect

    Hayes, A.; Johnson, W.C.

    1980-05-01

    Biological data of the Malaysian prawn, Macrobrachium rosenbergii, are summarized. A history on its rearing techniques is given, but through the use of geothermal water or industrial warm water effluent, its range can be expanded. The use of wasted geothermal water at the Oregon Institute of Technology for prawn ponds is noted. Pond management and design; the hatchery design and function for larval culture; and geothermal applications (legal aspects and constraints) are discussed. (MCW)

  7. Multi-culture solar heated bio-shelter. Final report

    SciTech Connect

    Not Available

    1985-01-01

    A rooftop greenhouse (bio-shelter) that is heated with active and passive solar systems is presented. The intent of the greenhouse is to grow vegetables hydroponically the year-round using a nutrient flow technique; and to growth the giant tropical Malaysian prawn Macrobrachium rosenbergii in a recycling raceway water system heated with solar power. The produce grown was continuously monitored and the harvests weighed in order to estimate the year-round production potential of the bio-shelter greenhouse.

  8. Organochlorine Pesticides and Biomarker Responses in Two Fishes Oreochromis niloticus (Linnaeus, 1758) and Chrysichthys nigrodigitatus (Lacepede, 1803) and an Invertebrate, Macrobrachium vollenhovenii (Herklot, 1857), from the Lake Taabo (Cote d`Ivoire)

    NASA Astrophysics Data System (ADS)

    Roche, Helene; Tidou, Abiba; Persic, Ana

    The concentrations of organochlorine pesticides (OCPs) were determined in muscle samples of two species of fish, tilapia (Oreochromis niloticus) and the catfish (Chrysichthys nigrodigitatus) and the prawn (Macrobrachium vollenhovenii) found in Lake Taabo (Cote d`Ivoire). Simultaneous measurements of enzymatic biomarkers were made to evaluate the ecotoxicological risk in this hydroelectric reservoir. Lindane and endosulfan were the dominant contaminants, suggesting their current use in neighboring agricultural areas. Other organochlorine (OC) compounds were detected, including some currently banned substances. Ranked in an order of descending concentrations, we found: DDT and its metabolites (17.8-57.2 ng g-1 dry weight), endrin (7.17-25.0 ng g-1 dry weight) and heptachlor (7.36-23.6 ng g-1 dry weight), as well as traces of isomers of chlordane, aldrin and fipronil. The hepatic Glutathione S-Transferase (GST) activity measured in fishes was not correlated with pesticide contamination; whereas the antioxidant biomarkers demonstrated some significant associations, especially hepatic catalase with lindane (R = 0.83) and Glutathione Peroxidase (GPx) with heptachlor epoxide (R = 0.84) and with pp`DDT (R = 0.81). In the prawns, acetylcholinesterase (AChE) activity showed significant negative correlations with DDT and its metabolites (R = -0.91). The results of this study emphasize the urgent need for overall environmental risk assessment studies in the region of Taabo and other developing areas.

  9. Gametogenesis in the freshwater prawn, Macrobrachium lamerrii.

    PubMed

    Sarojini, R; Gyananath, G

    1984-01-01

    Histological changes of the female and male gonads were studied and their relation to reproductive cycle is described. Ovary showed distinct changes during the course of its reproductive cycle. Maturational changes did not occur simultaneously in all follicles of the ovary. Regression of ovary during the non breeding season with a number of degenerating oocytes was observed. The seasonal changes in spermatogenesis revealed that growth in the testes appeared to be less than in the case of females. PMID:6741557

  10. Nodavirus Colonizes and Replicates in the Testis of Gilthead Seabream and European Sea Bass Modulating Its Immune and Reproductive Functions

    PubMed Central

    Valero, Yulema; Arizcun, Marta; Esteban, M. Ángeles; Bandín, Isabel; Olveira, José G.; Patel, Sonal; Cuesta, Alberto; Chaves-Pozo, Elena

    2015-01-01

    Viruses are threatening pathogens for fish aquaculture. Some of them are transmitted through gonad fluids or gametes as occurs with nervous necrosis virus (NNV). In order to be transmitted through the gonad, the virus should colonize and replicate inside some cell types of this tissue and avoid the subsequent immune response locally. However, whether NNV colonizes the gonad, the cell types that are infected, and how the immune response in the gonad is regulated has never been studied. We have demonstrated for the first time the presence and localization of NNV into the testis after an experimental infection in the European sea bass (Dicentrarchus labrax), and in the gilthead seabream (Sparus aurata), a very susceptible and an asymptomatic host fish species, respectively. Thus, we localized in the testis viral RNA in both species using in situ PCR and viral proteins in gilthead seabream by immunohistochemistry, suggesting that males might also transmit the virus. In addition, we were able to isolate infective particles from the testis of both species demonstrating that NNV colonizes and replicates into the testis of both species. Blood contamination of the tissues sampled was discarded by completely fish bleeding, furthermore the in situ PCR and immunocytochemistry techniques never showed staining in blood vessels or cells. Moreover, we also determined how the immune and reproductive functions are affected comparing the effects in the testis with those found in the brain, the main target tissue of the virus. Interestingly, NNV triggered the immune response in the European sea bass but not in the gilthead seabream testis. Regarding reproductive functions, NNV infection alters 17?-estradiol and 11-ketotestosterone production and the potential sensitivity of brain and testis to these hormones, whereas there is no disruption of testicular functions according to several reproductive parameters. Moreover, we have also studied the NNV infection of the testis in vitro to assess local responses. Our in vitro results show that the changes observed on the expression of immune and reproductive genes in the testis of both species are different to those observed upon in vivo infections in most of the cases. PMID:26691348

  11. Involvement of fish signal transducer and activator of transcription 3 (STAT3) in nodavirus infection induced cell death.

    PubMed

    Huang, Youhua; Huang, Xiaohong; Yang, Ying; Wang, Wei; Yu, Yepin; Qin, Qiwei

    2015-03-01

    The Janus kinase (JAK)-signal transducer and activator of transcription (STAT) signaling pathway is an important signaling pathway activated by interferons in response to virus infection. Fish STAT3 has been demonstrated to be involved in Singapore grouper iridovirus (SGIV) infection and virus induced paraptosis, but its effects on the replication of other fish viruses still remained uncertain. Here, the roles of grouper STAT3 (Ec-STAT3) in red spotted grouper nervous necrosis virus (RGNNV) infection were investigated. The present data showed that the distribution of phosphorylated Ec-STAT3 was altered in RGNNV infected fish cells, and the promoter activity of STAT3 was significantly increased during virus infection, suggesting that STAT3 activation was involved in RGNNV infection. Using STAT3 specific inhibitor, we found that inhibition of Ec-STAT3 in vitro did not affect the transcription and protein synthesis of RGNNV coat protein (CP), however, the severity of RGNNV induced vacuolation and autophagy was significantly increased. Meanwhile, at the late stage of virus infection, RGNNV induced necrotic cell death was significantly decreased after inhibition of Ec-STAT3. Further studies indicated that Ec-STAT3 inhibition significantly increased the transcript level of autophagy related genes, including UNC-51-like kinase 2 (ULK2) and microtubule-associated protein 1 light chain 3-II (LC3-II) induced by RGNNV infection. Moreover, the expression of several pro-inflammatory factors, including TNF?, IL-1? and IL-8 were mediated by Ec-STAT3 during RGNNV infection. Together, our results not only firstly revealed that STAT3 exerted novel roles in response to fish virus infection, but also provided new insights into understanding the roles of STAT3 in different forms of programmed cell death. PMID:25555814

  12. Immunochemical and immunocytochemical studies of the crustacean vitellogenesis-inhibiting hormone (VIH).

    PubMed

    Meusy, J J; Martin, G; Soyez, D; van Deijnen, J E; Gallo, J M

    1987-09-01

    Immunochemical investigations, using dot immunobinding assay (DIA) and enzyme-linked immunosorbent assay (ELISA), and immunocytochemical studies reveal the following new information about crustacean vitellogenesis-inhibiting hormone (VIH): (1) The structure of VIH is sufficiently different from that of the other sinus gland neuropeptides to allow a selective recognition of VIH by polyclonal antibodies. (2) From immunochemical criteria, VIH does not seem strictly species specific. The antisera raised against VIH of Homarus americanus cross-react with sinus gland extracts of Palaemonetes varians, Palaemon serratus, Macrobrachium rosenbergii, Carcinus maenas, and Porcellio dilatatus. (3) In the sinus gland of H. americanus, VIH immunoreactivity is localized mainly in electron-dense granules of medium size (110-185 nm in diameter) while, in P. dilatatus, the labeling is mostly on the largest granules (200-270 nm in diameter). PMID:3666410

  13. Permanent genetic resources added to molecular ecology resources database 1 December 2012-31 January 2013.

    PubMed

    Arranz, Silvia E; Avarre, Jean-Christophe; Balasundaram, Chellam; Bouza, Carmen; Calcaterra, Nora B; Cezilly, Frank; Chen, Shi-long; Cipriani, Guido; Cruz, V P; D'Esposito, D; Daniel, Carla; Dejean, Alain; Dharaneedharan, Subramanian; Díaz, Juan; Du, Man; Durand, Jean-Dominique; Dziadek, Jaros?aw; Foresti, F; Peng-cheng, Fu; Gao, Qing-bo; García, Graciela; Gauffre-Autelin, Pauline; Giovino, Antonio; Goswami, Mukunda; Guarino, Carmine; Guerra-Varela, Jorge; Gutiérrez, Verónica; Harris, D J; Heo, Moon-Soo; Khan, Gulzar; Kim, Mija; Lakra, Wazir S; Lauth, Jérémie; Leclercq, Pierre; Lee, Jeonghwa; Lee, Seung-Ho; Lee, Soohyung; Lee, Theresa; Li, Yin-hu; Liu, Hongbo; Liu, Shufang; Malé, Pierre-Jean G; Mandhan, Rishi Pal; Martinez, Paulino; Mayer, Veronika E; Mendel, Jan; Mendes, N J; Mendonça, F F; Minias, Alina; Minias, Piotr; Oh, Kyeong-Suk; Oliveira, C; Orivel, Jérôme; Orsini, L; Pardo, Belén G; Perera, A; Procaccini, G; Rato, C; Ríos, Néstor; Scibetta, Silvia; Sharma, Bhagwati S; Sierens, Tim; Singh, Akhilesh; Terer, Taita; Triest, Ludwig; Urbánková, So?a; Vera, Manuel; Villanova, Gabriela V; Voglmayr, Hermann; Vysko?ilová, Martina; Wang, Hongying; Wang, Jiu-li; Wattier, Rémi A; Xing, Rui; Yadav, Kamalendra; Yin, Guibo; Yuan, Yanjiao; Yun, Jong-Chul; Zhang, Fa-qi; Zhang, Jing-hua; Zhuang, Zhimeng

    2013-05-01

    This article documents the addition of 268 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Alburnoides bipunctatus, Chamaerops humilis, Chlidonias hybrida, Cyperus papyrus, Fusarium graminearum, Loxigilla barbadensis, Macrobrachium rosenbergii, Odontesthes bonariensis, Pelteobagrus vachelli, Posidonia oceanica, Potamotrygon motoro, Rhamdia quelen, Sarotherodon melanotheron heudelotii, Sibiraea angustata, Takifugu rubripes, Tarentola mauritanica, Trimmatostroma sp. and Wallago attu. These loci were cross-tested on the following species: Alburnoides fasciatus, Alburnoides kubanicus, Alburnoides maculatus, Alburnoides ohridanus, Alburnoides prespensis, Alburnoides rossicus, Alburnoides strymonicus, Alburnoides thessalicus, Alburnoides tzanevi, Carassius carassius, Fusarium asiaticum, Leucaspius delineatus, Loxigilla noctis dominica, Pelecus cultratus, Phoenix canariensis, Potamotrygon falkneri, Trachycarpus fortune and Vimba vimba. PMID:23521844

  14. Human health risk assessment of heavy metals in tropical fish and shellfish collected from the river Buriganga, Bangladesh.

    PubMed

    Ahmed, Md Kawser; Baki, Mohammad Abdul; Islam, Md Saiful; Kundu, Goutam Kumar; Habibullah-Al-Mamun, Md; Sarkar, Santosh Kumar; Hossain, Md Muzammel

    2015-10-01

    Although fish, crustacean, and shellfish are significant sources of protein, they are currently affected by rapid industrialization, resulting in increased concentrations of heavy metals. Accumulation of heavy metals (V, Cr, Mn, Ni, Cu, Zn, As, Se, Mo, Ag, Cd, Sb, Ba, and Pb) and associated human health risk were investigated in three fish species, namely Ailia coila, Gagata youssoufi, and Mastacembelus pancalus; one crustacean (prawn), Macrobrachium rosenbergii; and one Gastropoda, Indoplanorbis exustus, collected from the Buriganga River, Bangladesh. Samples were collected from the professional fishermen. Cu was the most accumulated metal in M. rosenbergii. Ni, As, Ag, and Sb were in relatively lower concentrations, whereas relatively higher accumulation of Cr, Mn, Zn, and Se were recorded. Mn, Zn, and Pb were present in higher concentrations than the guidelines of various authorities. There were significant differences in metal accumulation among different fish, prawn, or shellfish species. Target hazard quotient (THQ) and target cancer risk (TR) were calculated to estimate the non-carcinogenic and carcinogenic health risks, respectively. The THQ for individual heavy metals were below 1 suggesting no potential health risk. But combined impact, estimated by hazard index (HI), suggested health risk for M. pancalus consumption. Although consumption of fish at current accumulation level is safe but continuous and excess consumption for a life time of more than 70 years has probability of target cancer risk. PMID:26044144

  15. In-Depth Tanscriptomic Analysis on Giant Freshwater Prawns

    PubMed Central

    Mohd-Shamsudin, Maizatul Izzah; Kang, Yi; Lili, Zhao; Tan, Tian Tian; Kwong, Qi Bin; Liu, Hang; Zhang, Guojie; Othman, Rofina Yasmin; Bhassu, Subha

    2013-01-01

    Gene discovery in the Malaysian giant freshwater prawn (Macrobrachium rosenbergii) has been limited to small scale data collection, despite great interest in various research fields related to the commercial significance of this species. Next generation sequencing technologies that have been developed recently and enabled whole transcriptome sequencing (RNA-seq), have allowed generation of large scale functional genomics data sets in a shorter time than was previously possible. Using this technology, transcriptome sequencing of three tissue types: hepatopancreas, gill and muscle, has been undertaken to generate functional genomics data for M. rosenbergii at a massive scale. De novo assembly of 75-bp paired end Ilumina reads has generated 102,230 unigenes. Sequence homology search and in silico prediction have identified known and novel protein coding candidate genes (?24%), non-coding RNA, and repetitive elements in the transcriptome. Potential markers consisting of simple sequence repeats associated with known protein coding genes have been successfully identified. Using KEGG pathway enrichment, differentially expressed genes in different tissues were systematically represented. The functions of gill and hepatopancreas in the context of neuroactive regulation, metabolism, reproduction, environmental stress and disease responses are described and support relevant experimental studies conducted previously in M. rosenbergii and other crustaceans. This large scale gene discovery represents the most extensive transcriptome data for freshwater prawn. Comparison with model organisms has paved the path to address the possible conserved biological entities shared between vertebrates and crustaceans. The functional genomics resources generated from this study provide the basis for constructing hypotheses for future molecular research in the freshwater shrimp. PMID:23734171

  16. Effect of stocking density on extensive production of freshwater shrimp in coal mine reclamation ponds

    SciTech Connect

    Tidwell, J.H.; Wynne, F.; Coyle, S.D.; Grey, B.; McGuire, J.

    1998-12-31

    The use of post-mining reclamation ponds for the production of freshwater shrimp was evaluated by examining different stocking densities. Juvenile shrimp (Macrobrachium rosenbergii) averaging 0.5g each were stocked into four existing ponds at Peabody`s Ken Surface Mine at 6,175; 12,350; 18,500; and 24,700/ha (2,500, 5,000, 7,500, and 10,000/acre) on June 1, 1995. Shrimp were fed twice a week for 103 days, with harvest conducted September 13, 1995. Survival averaged 40%, overall. Average individual weight size was inversely related to stocking density ranging from 52 g (8.7 shrimp/lb) at 6,175/ha to 20.3 g (22.7 shrimp/lb) at 18,500/acre. Total production was directly related to stocking density ranging from 97 kg/ha (86 lbs/acre) at low density to 211 kg/ha (188 lbs/acre) at 18,500/ha shrimp acre. The major difficulty was at harvest due to difficulty in draining ponds. Construction of designed culture ponds with gravity drains during reclamation could greatly enhance survival, harvestability, and commercial feasibility.

  17. Isolation of AHL-degrading bacteria from micro-algal cultures and their impact on algal growth and on virulence of Vibrio campbellii to prawn larvae.

    PubMed

    Pande, Gde Sasmita Julyantoro; Natrah, Fatin Mohd Ikhsan; Flandez, Ace Vincent Bravo; Kumar, Uday; Niu, Yufeng; Bossier, Peter; Defoirdt, Tom

    2015-12-01

    Inactivation of quorum sensing (QS) signal molecules, such as acylhomoserine lactones (AHLs) of pathogenic bacteria, has been proposed as a novel method to combat bacterial diseases in aquaculture. Despite the importance of micro-algae for aquaculture, AHL degradation by bacteria associated with micro-algal cultures has thus far not been investigated. In this study, we isolated Pseudomonas sp. NFMI-T and Bacillus sp. NFMI-C from open cultures of the micro-algae Tetraselmis suecica and Chaetoceros muelleri, respectively. An AHL degradation assay showed that either monocultures or co-cultures of the isolates were able to degrade the AHL N-hexanoyl-L-homoserine lactone. In contrast, only Bacillus sp. NFMI-C was able to inactivate N-hydroxybutanoyl-L-homoserine lactone, the AHL produced by Vibrio campbellii. The isolated bacteria were able to persist for up to 3 weeks in conventionalized micro-algal cultures, indicating that they were able to establish and maintain themselves within open algal cultures. Using gnotobiotic algal cultures, we found that the isolates did not affect growth of the micro-algae from which they were isolated, whereas a mixture of both isolates increased the growth of Tetraselmis and decreased the growth of Chaetoceros. Finally, addition of Bacillus sp. NFMI-C to the rearing water of giant river prawn (Macrobrachium rosenbergii) larvae significantly improved survival of the larvae when challenged with pathogenic V. campbellii, whereas it had no effect on larval growth. PMID:26344339

  18. Stability of the WSSV ORF94 VNTR genotype marker during passage in marine shrimp, freshwater crayfish and freshwater prawns.

    PubMed

    Gudkovs, Nicholas; Murwantoko, Murwantoko; Walker, Peter J

    2014-10-16

    The white spot syndrome virus (WSSV) genome contains 3 variable number tandem repeat (VNTR) regions, located in open reading frame (ORF) 75, ORF94 and ORF125, which have been employed for molecular epizootiological studies. A previous report suggested that the ORF 94 VNTR is highly unstable, varying in the number of tandem repeats during single passages from shrimp to other crustaceans. As such rapid variations would have profound implications for the interpretation of molecular epizootiological data, we re-examined the stability of the ORF94 VNTR. Two WSSV isolates with different ORF94 VNTR genotypes (TRS5 and TRS7) were obtained from disease outbreaks in farmed black tiger shrimp Penaeus monodon in Indonesia. High titre stocks of each virus were produced by injection in specific pathogen-free (SPF) Pacific white shrimp Litopenaeus vannamei with filtered infected tissue extracts, and the genotypes were confirmed. Each stock (macerated tissue) was then used to feed SPF Pacific white shrimp, freshwater crayfish (Cherax sp.) and freshwater prawns Macrobrachium rosenbergii through 3 successive passages involving alternative hosts at each level. Taqman real-time PCR was conducted on samples from each group to confirm infection and quantify viral genetic loads. ORF94 VNTR genotype analysis conducted on samples from each of the 43 passage groups indicated no variations in the VNTR number in either genotype TRS5 or genotype TRS7. This finding is contrary to the previous report and suggests that ORF94 VNTR are stable during multiple passages in these crustaceans. PMID:25320037

  19. Evaluation of Trace Metal Content by ICP-MS Using Closed Vessel Microwave Digestion in Fresh Water Fish

    PubMed Central

    Jarapala, Sreenivasa Rao; Kandlakunta, Bhaskarachary; Thingnganing, Longvah

    2014-01-01

    The objective of the present study was to investigate trace metal levels of different varieties of fresh water fish using Inductively Coupled Plasma Mass Spectrophotometer after microwave digestion (MD-ICPMS). Fish samples were collected from the outlets of twin cities of Hyderabad and Secunderabad. The trace metal content in different varieties of analyzed fish were ranged from 0.24 to 1.68?mg/kg for Chromium in Cyprinus carpio and Masto symbollon, 0.20 to 7.52?mg/kg for Manganese in Labeo rohita and Masto symbollon, 0.006 to 0.07?mg/kg for Cobalt in Rastrelliger kanagurta and Pampus argenteus, 0.31 to 2.24?mg/kg for Copper in Labeo rohita and Penaeus monodon, 3.25 to 14.56?mg/kg for Zinc in Cyprinus carpio and Macrobrachium rosenbergii, and 0.01 to 2.05?mg/kg for Selenium in Rastrelliger kanagurta and Pampus argenteus, respectively. Proximate composition data for the different fishes were also tabulated. Since the available data for different trace elements for fish is scanty, here an effort is made to present a precise data for the same as estimated on ICP-MS. Results were in accordance with recommended daily intake allowance by WHO/FAO. PMID:24744789

  20. MAR 384 -DISEASES OF AQUATIC ORGANISMS (3 credits)

    E-print Network

    Allam, Bassem

    . Bivalve case studies: bacterial diseases (clam brown ring disease, juvenile oyster disease) 10. Bivalve (ISA, nodavirus, VHSV, IHNV, shrimp viruses, etc.) 13. Protozoan and Myxozoan parasites of crustaceans

  1. Transcript profiles of mitochondrial and cytoplasmic manganese superoxide dismutases in Exopalaemon carinicauda under ammonia stress

    NASA Astrophysics Data System (ADS)

    Ren, Hai; Li, Jian; Li, Jitao; Liu, Ping; Liang, Zhongxiu; Wu, Jianhua

    2015-05-01

    Superoxide dismutase (SOD) is one of the most important antioxidant defense enzymes, and is considered as the first line against oxidative stress. In this study, we cloned a mitochondrial manganese (Mn) SOD ( mMnSOD) cDNA from the ridgetail white prawn Exopalaemon carinicauda by using rapid amplification of cDNA ends (RACE) methods. The full-length cDNA for mMnSOD was 1 014-bp long, containing a 5'-untranslated region (UTR) of 37-bp, a 3'-UTR of 321-bp with a poly (A) tail, and included a 657-bp open reading frame encoding a protein of 218 amino acids with a 16-amino-acid signal peptide. The protein had a calculated molecular weight of 23.87 kDa and a theoretical isoelectric point of 6.75. The mMnSOD sequence included two putative N-glycosylation sites (NHT and NLS), the MnSOD signature sequence 180DVWEHAYY187, and four putative Mn binding sites (H48, H96, D180, and H184). Sequence comparison showed that the mMnSOD deduced amino acid sequence of E. carinicauda shared 97%, 95%, 89%, 84%, 82%, 72%, and 69% identity with that of Macrobrachium rosenbergii, Macrobrachium nipponense, Fenneropeneaus chinensis, Callinectes sapidus, Perisesarma bidens, Danio rerio, and Homo sapiens, resectively. Quantitative real-time RT-PCR analysis showed that mMnSOD transcripts were present in all E. carinicauda tissues examined, with the highest levels in the hepatopancreas. During an ammonia stress treatment, the transcript levels of mMnSOD and cMnSOD were up-regulated at 12 h in hemocytes and at 24 h in the hepatopancreas. As the duration of the ammonia stress treatment extended to 72 h, the transcript levels of mMnSOD and cMnSOD significantly decreased both in hemocytes and hepatopancreas. These findings indicate that the SOD system is induced to respond to acute ammonia stress, and may be involved in environmental stress responses in E. carinicauda.

  2. Molecular characterization of a cytosolic manganese superoxide dismutase from the Chinese mitten crab, Eriocheir sinensis.

    PubMed

    Zhao, D X; Chen, L Q; Qin, J G; Qin, C J; Zhang, H; Wu, P; Li, E C

    2014-01-01

    A cytosolic manganese superoxide dismutase gene (Es-cMnSOD) was cloned from the Chinese mitten crab Eriocheir sinensis, using reverse transcription-polymerase chain reaction and the rapid amplification of cDNA ends. The open reading frame of Es-cMnSOD is 867 bp in length and encodes a 288-amino acid protein without a signal peptide. The calculated molecular mass of the translated protein of Es-cMnSOD is 31.43 kDa, with an estimated isoelectric point of 6.30. The deduced amino acid sequence of Es-cMnSOD has similarities of 90, 89, 84, 87, and 81% to those of white shrimp Litopenaeus vannamei MnSOD, black tiger shrimp Penaeus monodon MnSOD, giant freshwater prawn Macrobrachium rosenbergii MnSOD, blue crab Callinectes sapidus MnSOD, and red swamp crayfish Procambarus clarkii MnSOD, respectively. Es-cMnSOD contains a manganese superoxide dismutase domain (DVWEHAYY) and 4 conserved amino acids responsible for binding manganese. Es-cMnSOD was expressed in the hemocytes, eyestalk, muscle, intestine, gill, and hepatopancreas. Es-cMnSOD transcripts in hemocytes of E. sinensis increased at 1.5 and 48 h after injection of Aeromonas hydrophila, indicating that the induction of the SOD system response occurred within a short period of time. This study suggests that MnSOD may play a critical role in crab immunity, allowing efficient activation of an early innate immune response in the crab. PMID:25501153

  3. Identification and cloning of a selenium-dependent glutathione peroxidase from tiger shrimp, Penaeus monodon, and its transcription following pathogen infection and related to the molt stages.

    PubMed

    Liu, Kuan-Fu; Yeh, Maw-Sheng; Kou, Guang-Hsiung; Cheng, Winton; Lo, Chu-Fang

    2010-09-01

    Complementary (c)DNA encoding glutathione peroxidase (GPx) messenger (m)RNA of the tiger shrimp Penaeus monodon was obtained from haemocytes by a reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) method. The 1321-bp cDNA contained an open reading frame (ORF) of 564bp, a 69-bp 5'-untranslated region (UTR), and a 688-bp 3'-UTR containing a poly A tail and a conserved selenocysteine insertion sequence (SECIS) element. The molecular mass of the deduced amino acid (aa) sequence (188 aa) was 21.05kDa long with an estimated pI of 7.68. It contains a putative selenocysteine residue which is encoded by the unusual stop codon, (190)TGA(192), and forms the active site with residues Glu(75) and Trp(143). Comparison of amino acid sequences showed that tiger shrimp GPx is more closely related to vertebrate GPx1, in accordance with those in Litopenaeus vannamei and Macrobrachium rosenbergii. GPx cDNA was synthesised in lymphoid organ, gills, heart, haemocytes, the hepatopancreas, muscles, and intestines. After injected with either Photobacterium damsela or white spot syndrome virus (WSSV), the respiratory bursts of shrimp significantly increased in order to kill the pathogen, and induced increases in the activities of superoxide dismutase and GPx, and regulation in the expression of cloned GPx mRNA to protect cells against damage from oxidation. The GPx expression significantly increased at stage D(0/1), and then gradually decreased until stage C suggesting that the cloned GPx might play a role in the molt regulation of shrimp. PMID:20399225

  4. Identification, Characterization, and Diel Pattern of Expression of Canonical Clock Genes in Nephrops norvegicus (Crustacea: Decapoda) Eyestalk.

    PubMed

    Sbragaglia, Valerio; Lamanna, Francesco; M Mat, Audrey; Rotllant, Guiomar; Joly, Silvia; Ketmaier, Valerio; de la Iglesia, Horacio O; Aguzzi, Jacopo

    2015-01-01

    The Norway lobster, Nephrops norvegicus, is a burrowing decapod with a rhythmic burrow emergence (24 h) governed by the circadian system. It is an important resource for European fisheries and its behavior deeply affects its availability. The current knowledge of Nephrops circadian biology is phenomenological as it is currently the case for almost all crustaceans. In attempt to elucidate the putative molecular mechanisms underlying circadian gene regulation in Nephrops, we used a transcriptomics approach on cDNA extracted from the eyestalk, a structure playing a crucial role in controlling behavior of decapods. We studied 14 male lobsters under 12-12 light-darkness blue light cycle. We used the Hiseq 2000 Illumina platform to sequence two eyestalk libraries (under light and darkness conditions) obtaining about 90 millions 100-bp paired-end reads. Trinity was used for the de novo reconstruction of transcriptomes; the size at which half of all assembled bases reside in contigs (N50) was equal to 1796 (light) and 2055 (darkness). We found a list of candidate clock genes and focused our attention on canonical ones: timeless, period, clock and bmal1. The cloning of assembled fragments validated Trinity outputs. The putative Nephrops clock genes showed high levels of identity (blastx on NCBI) with known crustacean clock gene homologs such as Eurydice pulchra (period: 47%, timeless: 59%, bmal1: 79%) and Macrobrachium rosenbergii (clock: 100%). We also found a vertebrate-like cryptochrome 2. RT-qPCR showed that only timeless had a robust diel pattern of expression. Our data are in accordance with the current knowledge of the crustacean circadian clock, reinforcing the idea that the molecular clockwork of this group shows some differences with the established model in Drosophila melanogaster. PMID:26524198

  5. Identification, Characterization, and Diel Pattern of Expression of Canonical Clock Genes in Nephrops norvegicus (Crustacea: Decapoda) Eyestalk

    PubMed Central

    Sbragaglia, Valerio; Lamanna, Francesco; M. Mat, Audrey; Rotllant, Guiomar; Joly, Silvia; Ketmaier, Valerio; de la Iglesia, Horacio O.; Aguzzi, Jacopo

    2015-01-01

    The Norway lobster, Nephrops norvegicus, is a burrowing decapod with a rhythmic burrow emergence (24 h) governed by the circadian system. It is an important resource for European fisheries and its behavior deeply affects its availability. The current knowledge of Nephrops circadian biology is phenomenological as it is currently the case for almost all crustaceans. In attempt to elucidate the putative molecular mechanisms underlying circadian gene regulation in Nephrops, we used a transcriptomics approach on cDNA extracted from the eyestalk, a structure playing a crucial role in controlling behavior of decapods. We studied 14 male lobsters under 12–12 light-darkness blue light cycle. We used the Hiseq 2000 Illumina platform to sequence two eyestalk libraries (under light and darkness conditions) obtaining about 90 millions 100-bp paired-end reads. Trinity was used for the de novo reconstruction of transcriptomes; the size at which half of all assembled bases reside in contigs (N50) was equal to 1796 (light) and 2055 (darkness). We found a list of candidate clock genes and focused our attention on canonical ones: timeless, period, clock and bmal1. The cloning of assembled fragments validated Trinity outputs. The putative Nephrops clock genes showed high levels of identity (blastx on NCBI) with known crustacean clock gene homologs such as Eurydice pulchra (period: 47%, timeless: 59%, bmal1: 79%) and Macrobrachium rosenbergii (clock: 100%). We also found a vertebrate-like cryptochrome 2. RT-qPCR showed that only timeless had a robust diel pattern of expression. Our data are in accordance with the current knowledge of the crustacean circadian clock, reinforcing the idea that the molecular clockwork of this group shows some differences with the established model in Drosophila melanogaster. PMID:26524198

  6. cDNA cloning, characterization and expression analysis of peroxiredoxin 5 gene in the ridgetail white prawn Exopalaemon carinicauda.

    PubMed

    Duan, Yafei; Liu, Ping; Li, Jitao; Li, Jian; Gao, Baoquan; Chen, Ping

    2013-12-01

    Peroxiredoxin is a superfamily of antioxidative proteins that play important roles in protecting organisms against the toxicity of reactive oxygen species. In this study, a full-length of peroxiredoxin 5 (designated EcPrx5) cDNA was cloned from the ridgetail white prawn Exopalaemon carinicauda by using rapid amplification of cDNA ends (RACE) approaches. The full-length cDNA of the EcPrx5 was of 827 bp, containing a 5' untranslated region (UTR) of 14 bp, a 3' UTR of 228 bp with a poly (A) tail, and an open reading frame of 585 bp encoding a polypeptide of 194 amino acids with the predicted molecular weight of 20.83 kDa and estimated isoelectric point of 7.62. BLAST analysis revealed that amino acids of EcPrx5 shared 89, 68, 66, 65, 53 and 51 % identity with that of Macrobrachium rosenbergii, Megachile rotundata, Harpegnathos saltator, Acromyrmex echinatior, Danio rerio, and Homo sapiens counterparts, respectively. The conserved Prx domain and the signature of peroxiredoxin catalytic center identified in EcPrx5 suggested that EcPrx5 belonged to the atypical 2-Cys Prx subgroup. Real time quantitative RT-PCR analysis indicated that EcPrx5 could be detected in all the tested tissues with highest expression level in hepatopancreas. As time progressed, the expression level of EcPrx5 both in hemocytes and hepatopancreas increased in the first 6 h after Vibrio anguillarum and white spot syndrome virus challenge, and showed different expression profiles. The results indicated that EcPrx5 involved in immune response against bacterial and viral infection in E. carinicauda. PMID:24141991

  7. FishMicrosat: a microsatellite database of commercially important fishes and shellfishes of the Indian subcontinent

    PubMed Central

    2013-01-01

    Background Microsatellite DNA is one of many powerful genetic markers used for the construction of genetic linkage maps and the study of population genetics. The biological databases in public domain hold vast numbers of microsatellite sequences for many organisms including fishes. The microsatellite data available in these data sources were extracted and managed into a database that facilitates sequences analysis and browsing relevant information. The system also helps to design primer sequences for flanking regions of repeat loci for PCR identification of polymorphism within populations. Description FishMicrosat is a database of microsatellite sequences of fishes and shellfishes that includes important aquaculture species such as Lates calcarifer, Ctenopharyngodon idella, Hypophthalmichthys molitrix, Penaeus monodon, Labeo rohita, Oreochromis niloticus, Fenneropenaeus indicus and Macrobrachium rosenbergii. The database contains 4398 microsatellite sequences of 41 species belonging to 15 families from the Indian subcontinent. GenBank of NCBI was used as a prime data source for developing the database. The database presents information about simple and compound microsatellites, their clusters and locus orientation within sequences. The database has been integrated with different tools in a web interface such as primer designing, locus finding, mapping repeats, detecting similarities among sequences across species, and searching using motifs and keywords. In addition, the database has the ability to browse information on the top 10 families and the top 10 species, through record overview. Conclusions FishMicrosat database is a useful resource for fish and shellfish microsatellite analyses and locus identification across species, which has important applications in population genetics, evolutionary studies and genetic relatedness among species. The database can be expanded further to include the microsatellite data of fishes and shellfishes from other regions and available information on genome sequencing project of species of aquaculture importance. PMID:24047532

  8. Characterization of a Prawn OA/TA Receptor in Xenopus Oocytes Suggests Functional Selectivity between Octopamine and Tyramine

    PubMed Central

    Jezzini, Sami H.; Reyes-Colón, Dalynés; Sosa, María A.

    2014-01-01

    Here we report the characterization of an octopamine/tyramine (OA/TA or TyrR1) receptor (OA/TAMac) cloned from the freshwater prawn, Macrobrachium rosenbergii, an animal used in the study of agonistic social behavior. The invertebrate OA/TA receptors are seven trans-membrane domain G-protein coupled receptors that are related to vertebrate adrenergic receptors. Behavioral studies in arthropods indicate that octopaminergic signaling systems modulate fight or flight behaviors with octopamine and/or tyramine functioning in a similar way to the adrenalins in vertebrate systems. Despite the importance of octopamine signaling in behavioral studies of decapod crustaceans there are no functional data available for any of their octopamine or tyramine receptors. We expressed OA/TAMac in Xenopus oocytes where agonist-evoked trans-membrane currents were used as readouts of receptor activity. The currents were most effectively evoked by tyramine but were also evoked by octopamine and dopamine. They were effectively blocked by yohimbine. The electrophysiological approach we used enabled the continuous observation of complex dynamics over time. Using voltage steps, we were able to simultaneously resolve two types of endogenous currents that are affected over different time scales. At higher concentrations we observe that octopamine and tyramine can produce different and opposing effects on both of these currents, presumably through the activity of the single expressed receptor type. The pharmacological profile and apparent functional-selectivity are consistent with properties first observed in the OA/TA receptor from the insect Drosophila melanogaster. As the first functional data reported for any crustacean OA/TA receptor, these results suggest that functional-selectivity between tyramine and octopamine is a feature of this receptor type that may be conserved among arthropods. PMID:25350749

  9. Abundance of Ohio shrimp (Macrobrachium ohione) and Glass shrimp (Palaemonetes kadiakensis) in the unimpounded Upper Mississippi River

    USGS Publications Warehouse

    Barko, V.A.; Hrabik, R.A.

    2004-01-01

    Large rivers of the United States have been altered by construction and maintenance of navigation channels, which has resulted in habitat loss and degradation. Using 7 y of Long Term Resource Monitoring Program data collected from the unimpounded upper Mississippi River, we investigated Ohio and Glass Shrimp abundance collected from four physical habitats of the unimpounded upper Mississippi River: main channel border, main channel border with wing dike, open side channel and closed side channel. Our objective was to assess associations between Ohio and Glass Shrimp abundance, environmental measurements and the four habitats to better understand the ecology of these species in a channelized river system. Ohio Shrimp were most abundant in the open side channels, while Glass Shrimp were most abundant in the main channel border wing dike habitat. Thirty-two percent of the variance in Glass Shrimp abundance was explained by year 1995, year 1998, water temperature, depth of gear deployment, Secchi disk transparency and river elevation. Approximately 8% of variation in Ohio Shrimp abundance was explained by Secchi disk transparency. Catch-per-unit-effort (CPUE) was greatest in 1998 for Glass Shrimp but lowest in 1997. Conversely, CPUE was greatest in 1996 for Ohio Shrimp and lowest in 2000. Both species exhibited inter-annual variability in CPUE. Long-term impacts of river modifications on aquatic invertebrates have not been well documented in many large, river systems and warrants further study. The findings from this study provide ecological information on Glass and Ohio Shrimp in a channelized river system.

  10. Cloning and analysis of antiviral activity of a barramundi (Lates calcarifer) Mx gene.

    PubMed

    Wu, Y C; Chi, S C

    2007-07-01

    We obtained a full-length cDNA clone for the Mx gene of barramundi (Lates calcarifer), using RACE (rapid amplification of cDNA ends) polymerase chain reaction (PCR) amplification of RNA extracted from a barramundi brain cell line cBB. The Mx cDNA of 2.2kb contains an open reading frame (ORF) of 1875 nucleotides encoding a protein of 624 amino acids. The predicted barramundi Mx protein is 71.4 kDa and contains a tripartite guanosinetriphosphate (GTP)-binding motif at the amino terminal and a leucine zipper at the carboxyl terminal, characteristic of all known Mx proteins. Poly I:C-transfection induced the expression of Mx gene in cBB cells, and the induction level at 28 degrees C was higher than that at 20 degrees C. Moreover, Mx gene expression was also induced by viral infection, including fish nodavirus, birnavirus, and iridovirus. Among these, nodavirus was a stronger inducer than the other two viruses. Using an antiviral activity assay, we revealed that poly I:C-transfected cBB cells had antiviral activity against fish nodavirus and birnavirus, but not iridovirus. Furthermore, the replication of nodavirus and birnavirus could be restored after the expression of Mx gene was down-regulated by siRNA. Therefore, these results indicated that the expression of barramundi Mx gene was able to inhibit the proliferation of fish nodavirus and birnavirus. PMID:17097891

  11. Supplement 22, Part 1, Authors: A To Z 

    E-print Network

    Zidar, Judith A.; Shaw, Judith H.; Hanfman, Deborah T.; Kirby, Margie D.; Rayburn, Jane D.; Edwards, Shirley J.; Hood, Martha W.

    1978-01-01

    of vertebrates to serve as paratenic host Angiostrongylus cantonensis Alicata, J. E.; and McCarthy, Deroceras laeve D. D., 1964 a Vaginalus plebeius all from island of Rarotonga Bradybaena similaris Subulina octona Macrobrachium sp. planarians Rattus...

  12. SPECIES INTERACTIONS BETWEEN ESTUARINE DETRITIVORES: INHIBITION OR FACILITATION?

    EPA Science Inventory

    Native Hawaiian estuarine detritivores; the prawn Macrobrachium grandimanus, and the neritid gastropod Neritina vespertina, were maintained in flow-through microcosms with conditioned leaves from two riparian tree species, Hau (Hibiscus tiliaceus) and guava (Psidium guajava). Th...

  13. Comparison of betanodavirus replication efficiency in ten Indian fish cell lines.

    PubMed

    Sarath Babu, V; Abdul Majeed, S; Nambi, K S N; Taju, G; Madan, N; Sundar Raj, N; Sahul Hameed, A S

    2013-06-01

    Ten cell lines established from Indian marine, brackishwater and freshwater fish were tested for their susceptibility to fish nodavirus. In addition, the efficiency of betanodavirus replication was tested in these cell lines. Multiple vacuolation, a typical cytopathic effect for virus infection, was observed in infected SISK, SISS, SIGE and ICF cells. Infection of the different fish cell lines was confirmed by RT-PCR, immunodot blot assay and indirect ELISA. The virus concentration in culture supernatant collected from infected sea bass and grouper cell lines increased progressively from 10(3) at day 1 postinfection to 10(8) TCID50 ml(-1) at day 9. The amount of virus in different cell lines was also quantified by real-time PCR. These results indicate the suitability of the SISK, SISS, and SIGE cell lines for fish nodavirus propagation for developing viral diagnostics and vaccines. PMID:23392632

  14. Persistent infection of betanodavirus in a novel cell line derived from the brain tissue of barramundi Lates calcarifer.

    PubMed

    Chi, S C; Wu, Y C; Cheng, T M

    2005-06-01

    In order to obtain an in vitro system for studying the mechanism of persistent infection of fish nodavirus, a novel cell line (BB) was established from the brain tissue of a barramundi, Lates calcarifer, which had survived viral nervous necrosis disease. The cell line has been subcultured > 100 times. The persistence of fish nodavirus designated as barramundi brain nervous necrosis virus (BBNNV) in the BB cells was demonstrated by: (1) the detection of the infectious virus in the culture supernatants, (2) the detection of NNV nucleic acids extracted from the BB cells, (3) the positive result of immunochemical staining using an NNV-specific monoclonal antibody and (4) their resistance to infection by another fish nodavirus grouper NNV (GNNV). No temperature-sensitive mutants were detected in the culture supernatant of the BB cells. Neither truncated genome (RNA1 or RNA2) nor smaller coat protein was found in the purified BBNNV particles, suggesting that defective interfering particles were unlikely to be important in the NNV-persistent infection in the BB cells. The result of the neutralization test indicated that the 5 antigenic determinants, recognized by GNNV-specific neutralizing antibodies, also existed on the coat protein of BBNNV. The BB cell line is the first cell line reported to be persistently infected with NNV, and would be a useful model for understanding the mechanisms of NNV-persistent infection in vitro and in vivo. PMID:16060261

  15. 'Source: Holthuis, L. B. FAO Species Catalogue: Shrimps and Prawns of the World. FIRls125 Vol. 1 Foreign Fishery Developments

    E-print Network

    , and phytoplankton are cultured to feed the larval shrimp. The PPFC operates a semi-intensive farm for M. rosenbergii production at Thanatpin. Semi-intensive culture tech- niques consist of moderate water ex- change rates (less characterize semi- intensive culture. In countries like Bur- ma, farmers using semi-intensive tech- niques

  16. zoogeography. Proc. of the Centenary and Bicentenary Congress of Biology. Singapore. 1958. p. 170-181.

    E-print Network

    of M. rosellber- gii. III J. A. Hanson and H. L. Goodwin leditors). Shrimp and prawn farming Macrobrachillm rosenbergii de Man. III Symposium on Crustacea. Part 1. p. 649-669 Mar. BioI. Assoc. India. Ernakulum. REESE. E. S. 1964. Ethology and marine zoology. Oceanogr. Mar. BioI. Annu. Rev. 2

  17. Two new mite species of the subfamily Harpirhynchinae Dubinin, 1957 (Acariformes: Harpirhynchidae), parasites of the passerine birds (Aves: Passeriformes) in Australia and South Asia.

    PubMed

    Bochkov, Andre V; Klompen, Hans

    2015-09-01

    Two new mite species of the subfamily Harpirhynchinae Dubinin, 1957 (Acariformes: Harpirhynchidae) are described from passerine birds (Aves: Passeriformes): Harpirhynchoides artamus n. sp. from Artamus fuscus Vieillot (Artamidae) from an unknown locality in South Asia and Neharpyrhynchus domrowi n. sp. from three host species of the family Meliphagidae, Acanthorhynchus tenuirostris (Latham) (type-host) from Australia (New South Walles), Ptiloprora perstriata (De Vis) and Myzomela rosenbergii Schlegel from Papua New Guinea. PMID:26249519

  18. Environmental factors influencing adult sex ratio in Trinidadian guppies.

    PubMed

    McKellar, Ann E; Turcotte, Martin M; Hendry, Andrew P

    2009-04-01

    Sex ratios can influence mating behaviour, population dynamics and evolutionary trajectories; yet the causes of natural sex ratio variation are often uncertain. Although secondary (birth) sex ratios in guppies (Poecilia reticulata) are typically 1:1, we recorded female-biased tertiary (adult) sex ratios in about half of our 48 samples and male-biased sex ratios in none of them. This pattern implies that some populations experience male-biased mortality, perhaps owing to variation in predation or resource limitation. We assessed the effects of predation and/or inter-specific resource competition (intraguild predation) by measuring the local catch-per-unit-effort (CPUE) of species (Rivulus killifish and Macrobrachium prawns) that may differentially prey on male guppies. We assessed the effects of resource levels by measuring canopy openness and algal biomass (chlorophyll a concentration). We found that guppy sex ratios were increasingly female-biased with increasing CPUE of Macrobrachium, and perhaps also Rivulus, and with decreasing canopy openness. We also found an interaction between predators and resource levels in that the effect of canopy openness was greatest when Macrobrachium CPUE was highest. Our study thus also reveals the value of simultaneously testing multiple environmental factors that may drive tertiary sex ratio variation. PMID:19132402

  19. GHSC70 Is Involved in the Cellular Entry of Nervous Necrosis Virus

    PubMed Central

    Chang, Jui-Shin

    2014-01-01

    ABSTRACT Nervous necrosis virus (NNV) is a devastating pathogen of cultured marine fish and has affected more than 40 fish species. NNV belongs to the betanodaviruses of Nodaviridae and is a nonenveloped icosahedral particle with 2 single-stranded positive-sense RNAs. To date, knowledge regarding NNV entry into the host cell remains limited, and no NNV-specific receptor protein has been published. Using grouper fin cell line GF-1 and purified NNV capsid protein in a virus overlay protein binding assay (VOPBA), grouper heat shock cognate protein 70 (GHSC70) and grouper voltage-dependent anion selective channel protein 2 (GVDAC2) were investigated as NNV receptor protein candidates. We cloned and sequenced the genes for GHSC70 and GVDAC2 and expressed them in Escherichia coli for antiserum preparation. Knockdown of the expression of GHSC70 and GVDAC2 genes with specific short interfering RNAs (siRNAs) significantly downregulated viral RNA expression in NNV-infected GF-1 cells. By performing an immunoprecipitation assay, we confirmed that GHSC70 interacted with NNV capsid protein, while VDAC2 did not. Immunofluorescence staining and flow cytometry analysis revealed the presence of the GHSC70 protein on the cell surface. After a blocking assay, we detected the NNV RNA2 levels after 1 h of adsorption to GF-1 cells; the level was significantly lower in the cells pretreated with the GHSC70 antiserum than in nontreated cells. Therefore, we suggest that GHSC70 participates in the NNV entry of GF-1 cells, likely functioning as an NNV receptor or coreceptor protein. IMPORTANCE Fish nodavirus has caused mass mortality of more than 40 fish species worldwide and resulted in huge economic losses in the past 20 years. Among the four genotypes of fish nodaviruses, the red-spotted grouper nervous necrosis virus (RGNNV) genotype exhibits the widest host range. In our previous study, we developed monoclonal antibodies with high neutralizing efficiency against grouper NNV in GF-1 cells, indicating that NNV-specific receptor(s) may exist on the GF-1 cell membrane. However, no NNV receptor protein has been published. In this study, we found GHSC70 to be an NNV receptor (or coreceptor) candidate through VOBPA and provided several lines of evidence demonstrating that GHSC70 protein has a role in the NNV entry step of GF-1 cells. To the best of our knowledge, this is the first report identifying grouper HSC70 and its role in NNV entry into GF-1 cells. PMID:25320288

  20. First report of viral nervous necrosis-induced mass mortality in hatchery-reared larvae of clownfish, Amphiprion sebae Bleeker.

    PubMed

    Binesh, C P; Renuka, K; Malaichami, N; Greeshma, C

    2013-12-01

    Frequent mortality was observed in the larval rearing facility of marine clownfish, Amphiprion sebae Bleeker. Mortality resulted in 80% loss of larval stock. Moribund larvae showed clinical signs typical of viral nervous necrosis, such as uncoordinated, corkscrew-like swimming behaviour, hypersensitivity to stimuli, darkening of body and assembly into large groups, similar to bunches of grapes. The aetiology of the disease was confirmed by gross observation of clinical signs, histopathology and molecular diagnosis. Histological studies revealed severe vacuolation in the brain and in the bipolar and ganglion layers of the eye. Molecular diagnosis by reverse transcription-polymerase chain reaction (RT-PCR) specific to piscine nodavirus yielded a positive result. The partial nucleotide sequences of the PCR-amplified fragment were 97-98% similar to other betanodavirus isolates reported globally and more closely aligned with red-spotted grouper nervous necrosis virus (RGNNV). This is the first report of susceptibility of clownfish, A. sebae, to betanodavirus and the presence of the RGNNV in India. PMID:24117972

  1. Rapid Evolution of Virus Sequences in Intrinsically Disordered Protein Regions

    PubMed Central

    Gitlin, Leonid; Hagai, Tzachi; LaBarbera, Anthony; Solovey, Mark; Andino, Raul

    2014-01-01

    Nodamura Virus (NoV) is a nodavirus originally isolated from insects that can replicate in a wide variety of hosts, including mammals. Because of their simplicity and ability to replicate in many diverse hosts, NoV, and the Nodaviridae in general, provide a unique window into the evolution of viruses and host-virus interactions. Here we show that the C-terminus of the viral polymerase exhibits extreme structural and evolutionary flexibility. Indeed, fewer than 10 positively charged residues from the 110 amino acid-long C-terminal region of protein A are required to support RNA1 replication. Strikingly, this region can be replaced by completely unrelated protein sequences, yet still produce a functional replicase. Structure predictions, as well as evolutionary and mutational analyses, indicate that the C-terminal region is structurally disordered and evolves faster than the rest of the viral proteome. Thus, the function of an intrinsically unstructured protein region can be independent of most of its primary sequence, conferring both functional robustness and sequence plasticity on the protein. Our results provide an experimental explanation for rapid evolution of unstructured regions, which enables an effective exploration of the sequence space, and likely function space, available to the virus. PMID:25502394

  2. On the relative importance of pool morphology and woody debris to distributions of shrimp in a Puerto Rican headwater stream

    USGS Publications Warehouse

    Pyron, M.; Covich, A.P.; Black, R.W.

    1999-01-01

    In this paper, we report the sizes and distributional orientation of woody debris in a headwater rainforest stream in the Luquillo Experimental Forest (LEF), Puerto Rico. We also provide results of a 4-month study of a wood addition experiment designed to increase cover for benthic macroinvertebrates (freshwater shrimp). We added branch-sized woody debris to 20 pools in three streams. We trapped four species of freshwater shrimp (two species of benthic detritivores and two predatory shrimp species) during each of the 4 months following wood additions. An analysis of pool morphology (maximum depth, surface area and volume) provided a useful predictor of shrimp abundances. In general, numbers of shrimps increased with sizes of stream pools. A repeated measures ANOVA demonstrated no effect of woody debris additions on total numbers of shrimp per pool area. Two detritivore species (Atya lanipes, a filter feeder and Xiphocaris elongata, a shredder) decreased in abundance with increased woody debris and there was no statistical relationship between woody debris additions and predators (Macrobrachium carcinus and M. crenulatum). Small woody debris additions may have altered flow velocities that were important to filter-feeding Atya at the microhabitat scale, although the overall velocities within pools were not altered by wood additions. Lower numbers of Atya and Xiphocaris in two of the three streams may result from the occurrence of two predaceous fishes (American eel and mountain mullet) and more predatory Macrobrachium in these streams. One likely interpretation of the results of this study is that the stream pools in these study reaches had sufficient habitat structure provided by numerous rock crevices (among large rocks and boulders) to provide refuge from predators. Addition of woody debris did not add significantly to the existing structure. These results may not apply to stream channels with sand and gravel substrata where crevices and undercut banks are lacking and where woody debris often plays a major role by providing structure and refuge.

  3. Molecular cloning, expression and functional analysis of ISG15 in orange-spotted grouper, Epinephelus coioides.

    PubMed

    Huang, Xiaohong; Huang, Youhua; Cai, Jia; Wei, Shina; Ouyang, Zhengliang; Qin, Qiwei

    2013-05-01

    Interferon-stimulated gene 15 (ISG15) is an ubiquitin homolog that is significantly induced by type I interferons or viral infections. Groupers, Epinephelus spp. being maricultured in China and Southeast Asian countries, always suffer from virus infection, including iridovirus and nodavirus. To date, the roles of grouper genes, especially interferon related genes in virus infection remained largely unknown. Here, the ISG15 homolog (EcISG15) was cloned from grouper Epinephelus coioides and its immune response to Singapore grouper iridovirus (SGIV) and grouper nervous necrosis virus (GNNV) was investigated. The full-length EcISG15 cDNA was composed of 948 bp and encoded a polypeptide of 155 amino acids with 37-68% identity with the known ISG15 homologs from other fish species. Amino acid alignment analysis indicated that EcISG15 contained two ubiquitin-like (UBL) domains and an Ub-conjugation domain (LRGG). Expressional analysis showed that EcISG15 was dramatically induced by GNNV infection, poly I:C or poly dA-dT treatment, but no obvious changes were observed during SGIV infection. Immunofluorescence assay showed that EcISG15 localized mainly in the cytoplasm of grouper cells in response to poly I:C stimulation or GNNV infection, but not in mock or SGIV infected cells. Western blot analysis indicated that the ISGylation was absent in SGIV-infected cells, but significantly enhanced in GNNV-infected or poly I:C transfected cells, suggesting that EcISG15 might play different roles in SGIV and GNNV infection. Furthermore, overexpression of EcISG15 in vitro inhibited the transcription of GNNV genes significantly. Taken together, the results indicated that fish ISG15 might exert important roles against RNA virus infection. PMID:23403156

  4. Identification of Novel Positive-Strand RNA Viruses by Metagenomic Analysis of Archaea-Dominated Yellowstone Hot Springs

    SciTech Connect

    Benjamin Bolduc; Daniel P. Shaughnessy; Yuri I. Wolf; Eugene V. Koonin; Francisco F. Roberto; Mark Young

    2012-05-01

    There are no known RNA viruses that infect Archaea. Filling this gap in our knowledge of viruses will enhance our understanding of the relationships between RNA viruses from the three domains of cellular life and, in particular, could shed light on the origin of the enormous diversity of RNA viruses infecting eukaryotes. We describe here the identification of novel RNA viral genome segments from high-temperature acidic hot springs in Yellowstone National Park in the United States. These hot springs harbor low-complexity cellular communities dominated by several species of hyperthermophilic Archaea. A viral metagenomics approach was taken to assemble segments of these RNA virus genomes from viral populations isolated directly from hot spring samples. Analysis of these RNA metagenomes demonstrated unique gene content that is not generally related to known RNA viruses of Bacteria and Eukarya. However, genes for RNA-dependent RNA polymerase (RdRp), a hallmark of positive-strand RNA viruses, were identified in two contigs. One of these contigs is approximately 5,600 nucleotides in length and encodes a polyprotein that also contains a region homologous to the capsid protein of nodaviruses, tetraviruses, and birnaviruses. Phylogenetic analyses of the RdRps encoded in these contigs indicate that the putative archaeal viruses form a unique group that is distinct from the RdRps of RNA viruses of Eukarya and Bacteria. Collectively, our findings suggest the existence of novel positive-strand RNA viruses that probably replicate in hyperthermophilic archaeal hosts and are highly divergent from RNA viruses that infect eukaryotes and even more distant from known bacterial RNA viruses. These positive-strand RNA viruses might be direct ancestors of RNA viruses of eukaryotes.

  5. Crystal structure of a nematode-infecting virus.

    PubMed

    Guo, Yusong R; Hryc, Corey F; Jakana, Joanita; Jiang, Hongbing; Wang, David; Chiu, Wah; Zhong, Weiwei; Tao, Yizhi J

    2014-09-01

    Orsay, the first virus discovered to naturally infect Caenorhabditis elegans or any nematode, has a bipartite, positive-sense RNA genome. Sequence analyses show that Orsay is related to nodaviruses, but molecular characterizations of Orsay reveal several unique features, such as the expression of a capsid-? fusion protein and the use of an ATG-independent mechanism for translation initiation. Here we report the crystal structure of an Orsay virus-like particle assembled from recombinant capsid protein (CP). Orsay capsid has a T = 3 icosahedral symmetry with 60 trimeric surface spikes. Each CP can be divided into three regions: an N-terminal arm that forms an extended protein interaction network at the capsid interior, an S domain with a jelly-roll, ?-barrel fold forming the continuous capsid, and a P domain that forms surface spike projections. The structure of the Orsay S domain is best aligned to T = 3 plant RNA viruses but exhibits substantial differences compared with the insect-infecting alphanodaviruses, which also lack the P domain in their CPs. The Orsay P domain is remotely related to the P1 domain in calicivirus and hepatitis E virus, suggesting a possible evolutionary relationship. Removing the N-terminal arm produced a slightly expanded capsid with fewer nucleic acids packaged, suggesting that the arm is important for capsid stability and genome packaging. Because C. elegans-Orsay serves as a highly tractable model for studying viral pathogenesis, our results should provide a valuable structural framework for further studies of Orsay replication and infection. PMID:25136116

  6. Photobacterium sanctipauli sp. nov. isolated from bleached Madracis decactis (Scleractinia) in the St Peter & St Paul Archipelago, Mid-Atlantic Ridge, Brazil

    PubMed Central

    Moreira, Ana Paula B.; Duytschaever, Gwen; Chimetto Tonon, Luciane A.; Fróes, Adriana M.; de Oliveira, Louisi S.; Amado-Filho, Gilberto M.; Francini-Filho, Ronaldo B.; De Vos, Paul; Swings, Jean; Thompson, Cristiane C.

    2014-01-01

    Five novel strains of Photobacterium (A-394T, A-373, A-379, A-397 and A-398) were isolated from bleached coral Madracis decactis (scleractinian) in the remote St Peter & St Archipelago (SPSPA), Mid-Atlantic Ridge, Brazil. Healthy M. decactis specimens were also surveyed, but no strains were related to them. The novel isolates formed a distinct lineage based on the 16S rRNA, recA, and rpoA gene sequences analysis. Their closest phylogenetic neighbours were Photobacterium rosenbergii, P. gaetbulicola, and P. lutimaris, sharing 96.6 to 95.8% 16S rRNA gene sequence similarity. The novel species can be differentiated from the closest neighbours by several phenotypic and chemotaxonomic markers. It grows at pH 11, produces tryptophane deaminase, presents the fatty acid C18:0, but lacks C16:0 iso. The whole cell protein profile, based in MALDI-TOF MS, distinguished the strains of the novel species among each other and from the closest neighbors. In addition, we are releasing the whole genome sequence of the type strain. The name Photobacterium sanctipauli sp. nov. is proposed for this taxon. The G + C content of the type strain A-394T (= LMG27910T = CAIM1892T) is 48.2 mol%. PMID:25024905

  7. Two Membrane-Associated Regions within the Nodamura Virus RNA-Dependent RNA Polymerase Are Critical for both Mitochondrial Localization and RNA Replication

    PubMed Central

    Gant, Vincent U.; Moreno, Stephanie; Varela-Ramirez, Armando

    2014-01-01

    ABSTRACT Viruses with positive-strand RNA genomes amplify their genomes in replication complexes associated with cellular membranes. Little is known about the mechanism of replication complex formation in cells infected with Nodamura virus. This virus is unique in its ability to lethally infect both mammals and insects. In mice and in larvae of the greater wax moth (Galleria mellonella), Nodamura virus-infected muscle cells exhibit mitochondrial aggregation and membrane rearrangement, leading to disorganization of the muscle fibrils on the tissue level and ultimately in hind limb/segment paralysis. However, the molecular basis for this pathogenesis and the role of mitochondria in Nodamura virus infection remains unclear. Here, we tested the hypothesis that Nodamura virus establishes RNA replication complexes that associate with mitochondria in mammalian cells. Our results showed that Nodamura virus replication complexes are targeted to mitochondria, as evidenced in biochemical, molecular, and confocal microscopy studies. More specifically, we show that the Nodamura virus RNA-dependent RNA polymerase interacts with the outer mitochondrial membranes as an integral membrane protein and ultimately becomes associated with functional replication complexes. These studies will help us to understand the mechanism of replication complex formation and the pathogenesis of Nodamura virus for mammals. IMPORTANCE This study will further our understanding of Nodamura virus (NoV) genome replication and its pathogenesis for mice. NoV is unique among the Nodaviridae in its ability to infect mammals. Here we show that NoV establishes RNA replication complexes (RCs) in association with mitochondria in mammalian cells. These RCs contain newly synthesized viral RNA and feature a physical interaction between mitochondrial membranes and the viral RNA-dependent RNA polymerase (RdRp), which is mediated by two membrane-associated regions. While the nature of the interaction needs to be explored further, it appears to occur by a mode distinct from that described for the insect nodavirus Flock House virus (FHV). The interaction of the NoV RdRp with mitochondrial membranes is essential for clustering of mitochondria into networks that resemble those described for infected mouse muscle and that are associated with fatal hind limb paralysis. This work therefore provides the first link between NoV RNA replication complex formation and the pathogenesis of this virus for mice. PMID:24696464

  8. On the Brain of a Crustacean: A Morphological Analysis of CaMKII Expression and Its Relation to Sensory and Motor Pathways

    PubMed Central

    Ammar, Dib; Nazari, Evelise M.

    2013-01-01

    Calcium/calmodulin kinase II (CaMKII) is a Ca2+-activated enzyme that is abundant in vertebrate and invertebrate brains. However, its characterization is poorly addressed in the nervous system of crustaceans, and, to our knowledge, no studies have determined the microanatomical location of CaMKII in a crustacean species. In this study, we found labeling of CaMKII in the eyestalk and brain of the prawn Macrobrachium acanthurus, by means of immunohistochemistry and Western blotting. Antibodies against neuron (ß tubulin III), glutamate receptor (GluA1), and FMRFamide were used in order to further characterize the CaMKII-labeled cells in the brain. In the eyestalk, strong labeling with CaMKII was observed in the photoreceptors. These cells, especially in the rhabdom, were also reactive to anti-ß tubulin III, whereas the pigment cells were labeled with anti-CaMKII. GluA1 co-located with CaMKII in the photoreceptors. Also, CaMKII appeared in the same sites as FMRFamide in the deutocerebrum, including the olfactory lobe, and in the tritocerebrum, specifically in the antennular neuropil, indicating that the synaptic areas in these regions may be related to sensory-motor processing. In the brain, the identification of cells and regions that express CaMKII contributes to the understanding of the processing of neural connections and the modulating role of CaMKII in decapod crustaceans. PMID:23741406

  9. Reduced transmission of human schistosomiasis after restoration of a native river prawn that preys on the snail intermediate host

    USGS Publications Warehouse

    Sokolow, Susanne H.; Huttinger, Elizabeth; Jouanard, Nicolas; Hsieh, Michael H.; Lafferty, Kevin D.; Kuris, Armand M.; Riveau, Gilles; Senghor, Simon; Thiam, Cheikh; D'Diaye, Alassane; Faye, Djibril Sarr; De Leo, Giulio A.

    2015-01-01

    Eliminating human parasitic disease often requires interrupting complex transmission pathways. Even when drugs to treat people are available, disease control can be difficult if the parasite can persist in nonhuman hosts. Here, we show that restoration of a natural predator of a parasite’s intermediate hosts may enhance drug-based schistosomiasis control. Our study site was the Senegal River Basin, where villagers suffered a massive outbreak and persistent epidemic after the 1986 completion of the Diama Dam. The dam blocked the annual migration of native river prawns (Macrobrachium vollenhoveni) that are voracious predators of the snail intermediate hosts for schistosomiasis. We tested schistosomiasis control by reintroduced river prawns in a before-after-control-impact field experiment that tracked parasitism in snails and people at two matched villages after prawns were stocked at one village’s river access point. The abundance of infected snails was 80% lower at that village, presumably because prawn predation reduced the abundance and average life span of latently infected snails. As expected from a reduction in infected snails, human schistosomiasis prevalence was 18 ± 5% lower and egg burden was 50 ± 8% lower at the prawn-stocking village compared with the control village. In a mathematical model of the system, stocking prawns, coupled with infrequent mass drug treatment, eliminates schistosomiasis from high-transmission sites. We conclude that restoring river prawns could be a novel contribution to controlling, or eliminating, schistosomiasis.                            

  10. Reduced transmission of human schistosomiasis after restoration of a native river prawn that preys on the snail intermediate host.

    PubMed

    Sokolow, Susanne H; Huttinger, Elizabeth; Jouanard, Nicolas; Hsieh, Michael H; Lafferty, Kevin D; Kuris, Armand M; Riveau, Gilles; Senghor, Simon; Thiam, Cheikh; N'Diaye, Alassane; Faye, Djibril Sarr; De Leo, Giulio A

    2015-08-01

    Eliminating human parasitic disease often requires interrupting complex transmission pathways. Even when drugs to treat people are available, disease control can be difficult if the parasite can persist in nonhuman hosts. Here, we show that restoration of a natural predator of a parasite's intermediate hosts may enhance drug-based schistosomiasis control. Our study site was the Senegal River Basin, where villagers suffered a massive outbreak and persistent epidemic after the 1986 completion of the Diama Dam. The dam blocked the annual migration of native river prawns (Macrobrachium vollenhoveni) that are voracious predators of the snail intermediate hosts for schistosomiasis. We tested schistosomiasis control by reintroduced river prawns in a before-after-control-impact field experiment that tracked parasitism in snails and people at two matched villages after prawns were stocked at one village's river access point. The abundance of infected snails was 80% lower at that village, presumably because prawn predation reduced the abundance and average life span of latently infected snails. As expected from a reduction in infected snails, human schistosomiasis prevalence was 18 ± 5% lower and egg burden was 50 ± 8% lower at the prawn-stocking village compared with the control village. In a mathematical model of the system, stocking prawns, coupled with infrequent mass drug treatment, eliminates schistosomiasis from high-transmission sites. We conclude that restoring river prawns could be a novel contribution to controlling, or eliminating, schistosomiasis. PMID:26195752

  11. Photobacterium panuliri sp. nov., an alkalitolerant marine bacterium isolated from eggs of spiny lobster, Panulirus penicillatus from Andaman Sea.

    PubMed

    Deep, Kamal; Poddar, Abhijit; Das, Subrata K

    2014-11-01

    A facultative anaerobe, alkalitolerant, gram-negative marine bacterium strain LBS5(T), was isolated from eggs carried on the pleopods of female spiny lobster (Panulirus penicillatus) in Andaman Sea from a depth of 3.5 m. Heterotrophic growth was observed at 15-38 °C and pH 5.5-11. Optimum growth occurred at 28 °C and pH 7.5. It can grow in the presence of 0.5-7 % NaCl (w/v), and the optimal NaCl required for growth was 2-4 %. 16S rRNA gene sequence analysis revealed the strain LBS5(T) belongs to the genus Photobacterium and showed 99.6 % similarity with P. aquae AE6(T), 98.2 % with P. aphoticum M46(T), 97 % with P. rosenbergii CC1(T), 96.9 % with P. lutimaris DF-42(T), and 96.6 % with P. halotolerans MACL01(T). The DNA-DNA similarities between strains LBS5(T) with other closely related strains were well below 70 %. The DNA G + C content was 50.52 (±0.9) mol%. The major fatty acids were C16:1w7c/w6c, C18:1w6c/w7c, C16:0, C15:0 iso, C16:0 10-methyl/17:1 iso w9c, C17:0 iso. Polar lipids included a phosphatidylglycerol, a diphosphatidylglycerol, a phosphatidylethanolamine, and one unidentified lipid. Based on the polyphasic evidences, strain LBS5(T) represents a novel species of the genus Photobacterium for which Photobacterium panuliri sp. nov. is proposed. The type strain is LBS5(T) (=DSM 27646(T) = LMG 27617(T) = JCM 19199(T)). PMID:24962598

  12. Updating the Vibrio clades defined by multilocus sequence phylogeny: proposal of eight new clades, and the description of Vibrio tritonius sp. nov.

    PubMed Central

    Sawabe, Tomoo; Ogura, Yoshitoshi; Matsumura, Yuta; Feng, Gao; Amin, AKM Rohul; Mino, Sayaka; Nakagawa, Satoshi; Sawabe, Toko; Kumar, Ramesh; Fukui, Yohei; Satomi, Masataka; Matsushima, Ryoji; Thompson, Fabiano L.; Gomez-Gil, Bruno; Christen, Richard; Maruyama, Fumito; Kurokawa, Ken; Hayashi, Tetsuya

    2013-01-01

    To date 142 species have been described in the Vibrionaceae family of bacteria, classified into seven genera; Aliivibrio, Echinimonas, Enterovibrio, Grimontia, Photobacterium, Salinivibrio and Vibrio. As vibrios are widespread in marine environments and show versatile metabolisms and ecologies, these bacteria are recognized as one of the most diverse and important marine heterotrophic bacterial groups for elucidating the correlation between genome evolution and ecological adaptation. However, on the basis of 16S rRNA gene phylogeny, we could not find any robust monophyletic lineages in any of the known genera. We needed further attempts to reconstruct their evolutionary history based on multilocus sequence analysis (MLSA) and/or genome wide taxonomy of all the recognized species groups. In our previous report in 2007, we conducted the first broad multilocus sequence analysis (MLSA) to infer the evolutionary history of vibrios using nine housekeeping genes (the 16S rRNA gene, gapA, gyrB, ftsZ, mreB, pyrH, recA, rpoA, and topA), and we proposed 14 distinct clades in 58 species of Vibrionaceae. Due to the difficulty of designing universal primers that can amplify the genes for MLSA in every Vibrionaceae species, some clades had yet to be defined. In this study, we present a better picture of an updated molecular phylogeny for 86 described vibrio species and 10 genome sequenced Vibrionaceae strains, using 8 housekeeping gene sequences. This new study places special emphasis on (1) eight newly identified clades (Damselae, Mediterranei, Pectenicida, Phosphoreum, Profundum, Porteresiae, Rosenbergii, and Rumoiensis); (2) clades amended since the 2007 proposal with recently described new species; (3) orphan clades of genomospecies F6 and F10; (4) phylogenetic positions defined in 3 genome-sequenced strains (N418, EX25, and EJY3); and (5) description of V. tritonius sp. nov., which is a member of the “Porteresiae” clade. PMID:24409173

  13. 210-Polonium studies in some environmental and biological matrices of Domiasiat uranium deposit area, West Khasi Hills, Meghalaya, India.

    PubMed

    Marbaniang, Deswyn G; Poddar, Raj K; Nongkynrih, Phlis; Khathing, Darlando T

    2010-03-01

    The study was performed using a silicon surface barrier alpha spectrometer at Bhabha Atomic Research Centre, Mumbai, India. Through the study, the observed (210)Po activity in water sample from different locations in the Domiasiat area ranges from 0.04 to 0.69 Bq/l. The daily and annual intake of (210)Po through water was also estimated and the mean value of 0.72 and 263.61 Bq, respectively, were observed. It is observed that the effective doses through water were higher than the World Health Organization recommended dose of 0.05 mSv/year. The total annual effective doses through terrestrial ingestion for all the locations was studied and the mean annual effective dose was observed to be 0.315 mSv, which, when compared to the worldwide and the Indian values, was observed to be slightly higher. The mean activity in soil is found to be 124.8 +/-5.7 Bq/kg and in meat the activity is 0.43 +/-0.05 Bq/kg. In fishes, an activity of 0.48 +/-0.07 Bq/kg in Garra lamta, 0.29 +/-0.02 Bq/kg in Neolissocheilus hexaganolepis, and 3.3 +/-0.1 Bq/kg in Macrobrachium sp. is observed. Activity concentration in plant samples was analyzed and the activity ranges from 0.020 +/-0.002 to 9.69 +/-0.35 Bq/kg. Committed effective dose by the adult population of the Domiasiat area through intake of (210)Po through these food items was also determined and compared with the Indian average value and the worldwide average value. PMID:19242810

  14. Use of a 15N tracer to determine linkages between a mangrove and an upland freshwater swamp

    NASA Astrophysics Data System (ADS)

    MacKenzie, R. A.; Cormier, N.

    2005-05-01

    Mangrove forests and adjacent upland freshwater swamps are important components of subsistence-based economies of Pacific islands. Mangroves provide valuable firewood (Rhizophora apiculata) and mangrove crabs (Scylla serrata); intact freshwater swamps are often used for agroforestry (e.g., taro cultivation). While these two systems are connected hydrologically via groundwater and surface flows, little information is available on how they may be biogeochemically or ecologically linked. For example, mangrove leaf litter was once thought to be an important food source for resident and transient nekton and invertebrates, but this value may have been overestimated. Instead, nutrients or allochthonous material (e.g., phytoplankton, detritus) delivered via groundwater or surface water from upland freshwater swamps may play a larger role in mangrove food webs. Understanding the linkages between these two ecologically and culturally important ecosystems will help us to understand the potential impacts of hydrological alterations that occur when roads or bridges are constructed through them. We conducted a 15N tracer study in the Yela watershed on the island of Kosrae, Federated States of Micronesia. K15NO3 was continually added at trace levels for 4 weeks to the Yela River in an upland freshwater swamp adjacent to a mangrove forest. Nitrate and ammonium pools, major primary producers, macroinvertebrates, and fish were sampled from stations 5 m upstream (freshwater swamp) and 138, 188, 213, and 313 m downstream (mangrove) from the tracer addition. Samples were collected once a week prior to, during, and after the 15N addition for a total of 6 weeks. Preliminary results revealed no significant enrichment (< 1 ‰) in the 15N isotope composition of either resident shrimp (Macrobrachium sp.) or mudskipper fish (Periophthalmus sp.). However, the 15N signature of ammonium pools was enriched 10-60 ‰ by the end of the third week. These results suggest that the tracer was present in the mangrove but was either unavailable to higher organisms or was incorporated into organic matter not utilized by shrimp or mudskippers.

  15. Carnivory during Ontogeny of the Plagioscion squamosissimus: A Successful Non-Native Fish in a Lentic Environment of the Upper Paraná River Basin.

    PubMed

    Neves, Mayara Pereira; Delariva, Rosilene Luciana; Guimarães, Ana Tereza Bittencourt; Sanches, Paulo Vanderlei

    2015-01-01

    This study evaluated feeding patterns and ontogenetic variations in a non-native fish species (Plagioscion squamosissimus) in an isolated lake in the Upper Paraná River floodplain. Quarterly samplings were performed from April 2005 to February 2006 using plankton nets to capture larvae, seining nets for juveniles, and gill nets and trammel for adults. Stomach contents (n = 378) were examined according to the volumetric method in which the volume of each food item was estimated using graduated test tubes or a glass counting plate. During early development (larval stage), P. squamosissimus consumed mainly Cladocera and Copepoda. Juveniles showed a more diverse diet, including shrimp (Macrobrachium amazonicum), fish, aquatic insects (Trichoptera, Ephemeroptera, Chironomidae and pupae of Diptera) and plants. It was notable the high proportion of cannibalism (23.3%) in this stage. Adults consumed predominantly shrimp and fish. The use of food resources varied significantly between development stages (ANOSIM; r = 0.458; p<0.005), showing changes in food preferences during ontogeny. The Similarity Percentage Analysis (SIMPER) indicated that Cladocera and Copepoda were responsible for the differences observed between the larval stages of pre-flexion, flexion and post-flexion. M. amazonicum and Chironomidae were responsible for the differences between juvenile and larval stages, while M. amazonicum and other fishes caused the differences between adults and other ontogenetic stages. These results are confirmed by the relationship between standard length and developmental periods (ANCOVA; r2 = 0.94; p<0.0001). In general, there were low values of trophic niche breadth. The essentially carnivorous habit from the early stages of P. squamosissimus and the predominant use of M. amazonicum by adults have important roles in feeding patterns of the species, suggesting a major contribution to its success and establishment, especially in lentic environments. PMID:26524336

  16. Carnivory during Ontogeny of the Plagioscion squamosissimus: A Successful Non-Native Fish in a Lentic Environment of the Upper Paraná River Basin

    PubMed Central

    Neves, Mayara Pereira; Delariva, Rosilene Luciana; Guimarães, Ana Tereza Bittencourt; Sanches, Paulo Vanderlei

    2015-01-01

    This study evaluated feeding patterns and ontogenetic variations in a non-native fish species (Plagioscion squamosissimus) in an isolated lake in the Upper Paraná River floodplain. Quarterly samplings were performed from April 2005 to February 2006 using plankton nets to capture larvae, seining nets for juveniles, and gill nets and trammel for adults. Stomach contents (n = 378) were examined according to the volumetric method in which the volume of each food item was estimated using graduated test tubes or a glass counting plate. During early development (larval stage), P. squamosissimus consumed mainly Cladocera and Copepoda. Juveniles showed a more diverse diet, including shrimp (Macrobrachium amazonicum), fish, aquatic insects (Trichoptera, Ephemeroptera, Chironomidae and pupae of Diptera) and plants. It was notable the high proportion of cannibalism (23.3%) in this stage. Adults consumed predominantly shrimp and fish. The use of food resources varied significantly between development stages (ANOSIM; r = 0.458; p<0.005), showing changes in food preferences during ontogeny. The Similarity Percentage Analysis (SIMPER) indicated that Cladocera and Copepoda were responsible for the differences observed between the larval stages of pre-flexion, flexion and post-flexion. M. amazonicum and Chironomidae were responsible for the differences between juvenile and larval stages, while M. amazonicum and other fishes caused the differences between adults and other ontogenetic stages. These results are confirmed by the relationship between standard length and developmental periods (ANCOVA; r2 = 0.94; p<0.0001). In general, there were low values of trophic niche breadth. The essentially carnivorous habit from the early stages of P. squamosissimus and the predominant use of M. amazonicum by adults have important roles in feeding patterns of the species, suggesting a major contribution to its success and establishment, especially in lentic environments. PMID:26524336

  17. Identification and molecular characterization of a peritrophin-like gene, involved in the antibacterial response in Chinese mitten crab, Eriocheir sinensis.

    PubMed

    Huang, Ying; Ma, FuTong; Wang, Wen; Ren, Qian

    2015-06-01

    Peritrophin was first isolated from insect peritrophic membrane (PM) and was thought to protect insects from invasion of microorganisms and to stimulate digestion of food. In this study, a peritrophin-like gene (EsPT) was obtained from Eriocheir sinensis. The full length cDNA of EsPT was 1232 bp, which contained 1005 bp ORF encoding a protein of 334 amino acids, including a 22 amino acid signal peptide, and 3 conserved chitin binding type 2 domains (ChtBD2) characterized by having a 6-cysteine motif. Phylogenetic analysis showed that EsPT was clustered together with 2 insect peritrophin-44-like proteins (MdP44L from Musca domestica and CcP44L from Ceratitis capitata), an insect chitin binding peritrophin-A domain containing protein (CfPT from Coptotermes formosanus) and a crustacean peritrophin (MnPT from Macrobrachium nipponense). Tissue distribution analysis revealed that EsPT was mainly expressed in hepatopancreas, intestine and hemocytes. The expression of EsPT is regulated by lipopolysaccharide, peptidoglycan, Staphylococcus aureus, Vibrio parahaemolyticus and Aeromonas hydrophila challenge. The recombinant EsPT could bind to different microbes, and enhanced the clearance of V. parahaemolyticus in vivo. In crabs, silencing of EsPT by siRNA suppressed the elimination of V. parahaemolyticus and increasing number of bacteria, finally upregulated the expression of anti-lipopolysaccharide factor (ALF) and clip domain serine proteases (cSP). The results might indicate that EsPT was involved in the anti-bacterial innate immunity of crabs. PMID:25596425

  18. Environmental occurrence and biota concentration of phthalate esters in Epe and Lagos Lagoons, Nigeria.

    PubMed

    Adeogun, Aina O; Ibor, Oju R; Omogbemi, Emmanuel D; Chukwuka, Azubuike V; Adegbola, Rachel A; Adewuyi, Gregory A; Arukwe, Augustine

    2015-07-01

    The high global occurrence of phthalates in different environmental matrixes has resulted in the detection of their metabolites in human urine, blood, and breast milk, indicating a widespread human exposure. In addition, the notorious endocrine disrupting effects of phthalates have shown that they mimic or antagonize the action of endogenous hormones, consequently producing adverse effects on reproduction, growth and development. Herein, we have studied the occurrence of phthalate esters (PEs) in water, sediment and biota of two lagoons (Epe and Lagos) in Nigeria. Two fish species (Tilapia guineensis, and Chrysichthys nigrodigitatus) and a crustacean (the African river prawn--Macrobrachium vollenhovenii) were analyzed for PEs levels using a HPLC method and the derived values were used for calculating bioconcentration factor (BCF), biota-sediment accumulation factor (BSAF) and phthalate pollution index (PPI) in the biota and environment. We observed that the growth and health condition of the fish species were normal with a k-factor of >1. Sediment PE levels were compared with water, at both lagoons showing concentration pattern that is characterized as DEHP = DEP > DBP. We observed that DBP was the predominant compound in T. guineensis, C. nigrodigitatus and African prawn, at both lagoons, showing organ-specific differences in bioconcentration (BCF and BSAF) patterns in the fish species. While there were no observed consistency in the pattern of PE concentration in fish organs, elevated DBP levels in different fish organs may be related to fish habitat and degradation level of phthalates. Low concentration of DEHP, compared with DBP and DEP, was measured in fish organs and whole prawn body. The BSAF values for DEHP were lowest, and highest for DBP for all species at both lagoons, and DEHP easily accumulated more in the sediment (sediment PPI = 0.28 and 0.16 for Epe and Lagos lagoon, respectively). Overall, our findings suggest a broader environmental and human health implication of the high PE levels in these lagoons since they represent significant sources of aquatic food resources for the neighboring communities. PMID:25935094

  19. Distribution and accumulation of polycyclic aromatic hydrocarbons (PAHs) in the food web of Nansi Lake, China.

    PubMed

    Zhang, Guizhai; Pan, Zhaoke; Wang, Xiaoming; Mo, Xiaojie; Li, Xiaoming

    2015-04-01

    The concentrations of polycyclic aromatic hydrocarbons (PAHs) were analyzed in water, sediment, and biota (aquatic plant, shrimp, and fish) of Nansi Lake by gas chromatography-mass spectrometry (GC-MS). The concentrations of total PAHs were 27.54-55.04 ng L(-1) in water, 80.31-639.23 ng g(-1) dry weight (dw) in sediments, 20.92-192.78 ng g(-1) dw in aquatic plants, and 67.3-533.9 ng g(-1) dw in fish and shrimp muscles. The ratios of phenanthrene to anthracene (Ph/An), fluoranthene to pyrene (Flu/Pyr), and low molecular weight to high molecular weight (LMW/HMW) in sediment indicated that the sources of the PAHs were a mixture of pyrolytic and petrogenic contamination at most sampling sites in Nansi Lake. The composition profile of PAHs in plants was similar to that in water and animals with 2-3 ring PAHs being dominant. The 4-6 ring PAHs were the dominant PAH compounds in sediment. There is a positive correlation between sediment and aquatic plants, but their PAH composition profiles were different, implying that aquatic plant absorption of PAHs from sediment is selective and the accumulation of PAHs in aquatic plants is different. The concentration of PAHs in fish showed a positive correlation with plants, reflecting that the PAHs in fish are mainly absorbed from aquatic plants rather than directly from the water. Bioaccumulation of LMW PAHs in aquatic biota was higher than HMW PAHs. The biota-sediment accumulation factor (BSAF) values of total PAHs in the plants Potamogeton lucens Linn and Ceratophyllum demersum Linn were higher than that in most animals. The BSAF values of total PAHs in animals were in the following order: Cyprinus carpio>Macrobrachium nipponense>Carassius auratus>Channa argus. There was no significant relationship between PAH bioaccumulation and trophic levels in Nansi Lake. Risk assessment of PAHs in water, sediment, and animals indicated that the water environment of Nansi Lake is safe at present. It is worthwhile to note that benzo [a] anthracene (BaA), benzo [a] pyrene (BaP), indeno [1,2,3-cd] pyrene (InP), dibenz [a, h] anthracene (DBA), and benzo [ghi] perylene (BghiP) were detected in sediment, plants, and animals at all sampling sites, and they have potential carcinogenicity to the organisms of Nansi Lake. PMID:25762425