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Sample records for macrobrachium rosenbergii nodavirus

  1. Developing immunological methods for detecting Macrobrachium rosenbergii nodavirus and extra small virus using a recombinant protein preparation.

    PubMed

    Wang, C-S; Chang, C-Y; Wen, C-M

    2016-06-01

    Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) have been identified as the causative agents for white tail disease (WTD) of M. rosenbergii. In this study, the gene sequences encoding MrNV and XSV capsid proteins were separately ligated into the pGEX-4T-3 expression vector and transformed into Escherichia coli. After induction, glutathione-S-transferase (GST)-tagged MrNV and XSV fusion proteins were obtained with molecular masses of 68 and 43 kDa, respectively. Specific polyclonal antibodies for MrNV and XSV against viral recombinant proteins and infected prawn tissues were verified using Western blotting. According to immunodot blot assay results, the detection sensitivities of antibodies were approximately 5 ng μL(-1) for both recombinant proteins GST-MrNV and GST-XSV. In additional, MrNV and XSV were detected at dilution levels of 1:2560 and 1:640 in the infected prawn tissues, respectively. No cross-reactions with white spot syndrome virus or grouper nervous necrosis virus were observed using immunodot blot assays. MrNV and XSV in infected muscle tissues were detected using immunohistochemistry. Although the detection limit of the immunodot blot assay was lower than that of nested reverse transcription polymerase chain reaction, these polyclonal antibodies can be useful for confirming MrNV and XSV infections in field tests. PMID:26263892

  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. Feasibility of direct utilization of selected geothermal water for aquaculture of macrobrachium rosenbergii. Technical report

    SciTech Connect

    Spinosa, C.

    1984-05-01

    The feasibility was tested of direct utilization of geothermal water for the aquaculture of Malaysian freshwater prawns (Macrobrachium rosenbergii). A problem with using geothermal water for aquaculture is the chemical composition of the water with high flouride levels being a particular problem. Results show that (1) some geothermal water in Idaho can be used directly for the aquaculture of Macrobrachium rosenbergii, (2) high flouride levels cannot be directly correlated with high mortality rates and (3) low flouride levels do not correlate with high growth rates.

  4. Routine Metabolic Rate and Limiting Oxygen Concentration of Freshwater Prawn Macrobrachium rosenbergii Larvae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Malaysian prawns, Macrobrachium rosenbergii, are hatched and raised indoors in small tanks. Prawns may be raised and shipped at high densities which could result in low dissolved oxygen (DO) conditions. Because DO may play an important role in prawn development and survival, we measured routine me...

  5. The effect of copper nanoparticles supplementation on freshwater prawn Macrobrachium rosenbergii post larvae.

    PubMed

    Muralisankar, Thirunavukkarasu; Saravana Bhavan, Periyakali; Radhakrishnan, Subramanian; Seenivasan, Chandirasekar; Srinivasan, Veeran

    2016-03-01

    The present study was performed to assess the effects of dietary supplementation of copper nanoparticles (Cu-NPs) on growth, biochemical constituents, digestive enzyme activities, antioxidant, metabolic enzyme levels, and non specific immune response of the freshwater prawn, Macrobrachium rosenbergii post larvae (PL). The Cu-NPs (200nm) were synthesized by facile and environmental friendly hydrothermal method. Cu-NPs were supplemented at 0, 10, 20, 40, 60, and 80mgkg(-1) with the basal diets. These Cu-NPs supplemented diets were fed to M. rosenbergii PL for 90 days. Results showed significant (P<0.05) improvements were observed in survival, growth, digestive enzyme activities, concentrations of biochemical constituents and total and differential haemocytes count of prawns fed with 20mgCu-NPskg(-1) supplemented feed. Prawns fed with 40-80mgCu-NPskg(-1) supplemented feed showed negative performance. Activity of antioxidants and metabolic enzymes in the muscle and hepatopancreas of prawns showed no significant alterations (P>0.05) prawns fed with up to 20mgCu-NPskg(-1) supplemented feeds. Whereas, prawns fed with 40-80mgCu-NPskg(-1) supplemented feed showed significant (P<0.05) elevations in antioxidant and metabolic enzymes activities. Hence, 40-80mgCu-NPskg(-1) diets may have toxic effect to M. rosenbergii. Hence, present study suggests that 20mgCu-NPskg(-1) can be supplemented for regulating better survival, growth and immune response of M. rosenbergii PL. PMID:26854244

  6. 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

  7. Genetic diversity of cultured and wild populations of the freshwater prawn Macrobrachium rosenbergii based on microsatellite analysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Freshwater prawn Macrobrachium rosenbergii culture in the Western Hemisphere is primarily, if not entirely, based on thirty-six individual prawn introduced to Hawaii from Malaysia in 1965 and 1966. Little information is available regarding the genetic background or current population status of cult...

  8. A transcriptome study on Macrobrachium rosenbergii hepatopancreas experimentally challenged with white spot syndrome virus (WSSV).

    PubMed

    Rao, Rama; Bhassu, Subha; Bing, Robin Zhu Ya; Alinejad, Tahereh; Hassan, Sharifah Syed; Wang, Jun

    2016-05-01

    The world production of shrimp such as the Malaysian giant freshwater prawn, Macrobrachium rosenbergii is seriously affected by the white spot syndrome virus (WSSV). There is an urgent need to understand the host pathogen interaction between M. rosenbergii and WSSV which will be able to provide a solution in controlling the spread of this infectious disease and lastly save the aquaculture industry. Now, using Next Generation Sequencing (NGS), we will be able to capture the response of the M. rosenbergii to the pathogen and have a better understanding of the host defence mechanism. Two cDNA libraries, one of WSSV-challenged M. rosenbergii and a normal control one, were sequenced using the Illumina HiSeq™ 2000 platform. After de novo assembly and clustering of the unigenes from both libraries, 63,584 standard unigenes were generated with a mean size of 698bp and an N50 of 1137bp. We successfully annotated 35.31% of all unigenes by using BLASTX program (E-value <10-5) against NCBI non-redundant (Nr), Swiss-Prot, Kyoto Encyclopedia of Genes and Genome pathway (KEGG) and Orthologous Groups of proteins (COG) databases. Gene Ontology (GO) assessment was conducted using BLAST2GO software. Differentially expressed genes (DEGs) by using the FPKM method showed 8443 host genes were significantly up-regulated whereas 5973 genes were significantly down-regulated. The differentially expressed immune related genes were grouped into 15 animal immune functions. The present study showed that WSSV infection has a significant impact on the transcriptome profile of M. rosenbergii's hepatopancreas, and further enhanced the knowledge of this host-virus interaction. Furthermore, the high number of transcripts generated in this study will provide a platform for future genomic research on freshwater prawns. PMID:26880158

  9. Identification and function of 11 Rab GTPases in giant freshwater prawn Macrobrachium rosenbergii.

    PubMed

    Huang, Ying; Ren, Qian

    2015-03-01

    Rab GTPases, members of the Ras-like GTPase superfamily, are central elements in endocytic membrane trafficking. However, little is known of the Rab genes in the giant freshwater prawn Macrobrachium rosenbergii. In this study, 11 Rab genes were identified from M. rosenbergii. All MrRabs have a RAB domain. Phylogenetic analysis showed that these 11 MrRabs were divided into different groups. The MrRab genes were ubiquitously expressed in heart, hemocytes, hepatopancreas, gills, stomach, and intestines. Real-time polymerase chain reaction revealed that the MrRab genes were significantly upregulated by white spot syndrome virus (WSSV) in the prawns, indicating that MrRabs might play an important role in innate immune response against WSSV. Moreover, after challenge with Vibrio parahaemolyticus, the expression levels of all MrRabs in the hepatopancreas were also upregulated, which might indicated the involvement of MrRabs in prawns antibacterial immunity. In all, these preliminary results showed that MrRabs were involved in innate immunity of M. rosenbergii. PMID:25542378

  10. 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

  11. Steroids and genes related to steroid biosynthesis in the female giant freshwater prawn, Macrobrachium rosenbergii.

    PubMed

    Thongbuakaew, Tipsuda; Siangcham, Tanapan; Suwansa-Ard, Saowaros; Elizur, Abigail; Cummins, Scott F; Sobhon, Prasert; Sretarugsa, Prapee

    2016-03-01

    The giant freshwater prawn, Macrobrachium rosenbergii, is important to many Asian countries due to its high economic value as an aquaculture product. With demand increasing, there is requirement for a better understanding of the biosynthetic components that regulate its growth and reproduction, including steroids, in order to help increase production. Vertebrate-type steroids and their receptors were identified in crustaceans and implicated in reproduction. In this study, we presented the sex steroids estradiol and progesterone by LC-MS/MS in female M. rosenbergii, and reveal steroidogenic-related genes by in silico analysis of de novo assembled transcriptomes. Comparative analysis with other species was performed to confirm their putative role, as well as tissue-specific and quantitative gene expression. We reveal 29 transcripts that encode for steroidogenic-related proteins, including steroidogenic enzymes, a nuclear steroid hormone receptors, and a steroidogenic factor. Moreover, we identified for the first time the presence of steroidogenic factor 1, StAR-related lipid transfer protein, estradiol receptor- and progesterone-like protein in M. rosenbergii. Those targeted for gene expression analysis (3 beta-hydroxysteroid dehydrogenase, 17 beta-hydroxysteroid dehydrogenase, estrogen sulfotransferase and progesterone receptor-like) showed widespread expression within many tissues, and at relatively high levels in the central nervous system (CNS) during ovarian maturation. In summary, we provide further evidence for the existence of steroidogenic pathways in crustaceans, which may be useful for advancing prawn aquaculture. PMID:26774430

  12. Isolation and Characterization of a Novel Dicistrovirus Associated with Moralities of the Great Freshwater Prawn, Macrobrachium rosenbergii

    PubMed Central

    Pan, Xiaoyi; Cao, Zheng; Yuan, Junfa; Shi, Zhengli; Yuan, Xuemei; Lin, Lingyun; Xu, Yang; Yao, Jiayun; Hao, Guijie; Shen, Jinyu

    2016-01-01

    The giant freshwater prawn, Macrobrachium rosenbergii, is an economically important crustacean and is farmed in many countries. Since 2009, a larval mortality syndrome of M. rosenbergii has broken out and spread widely in the main breeding area, including Zhejiang, Jiangsu, Guangxi, and Guangdong Provinces in mainland China. A novel virus, named Macrobrachium rosenbergii Taihu virus (MrTV), was isolated from the moribund larvae and was determined to be the causative agent of the M. rosenbergii larval mortality syndrome by experimental infection. Further genomic sequencing suggested that the MrTV genome is monopartite, 10,303 nt in length, and dicistronic with two non-overlapping open reading frames (ORFs) separated by an intergenic region (IGR) and flanked by untranslated regions (UTRs). Phylogenetic analysis using the full-length genomic sequence and the putative amino acid sequences of the capsid protein revealed that MrTV was more closely related to the taura syndrome virus (TSV) than to any other viruses. According to these molecular features, we proposed that MrTV is a new species in the genus Aparavirus, family Dicistroviridae. These results may shed light on controlling larval mortality syndrome in M. rosenbergii. PMID:26848659

  13. Isolation and Characterization of a Novel Dicistrovirus Associated with Moralities of the Great Freshwater Prawn, Macrobrachium rosenbergii.

    PubMed

    Pan, Xiaoyi; Cao, Zheng; Yuan, Junfa; Shi, Zhengli; Yuan, Xuemei; Lin, Lingyun; Xu, Yang; Yao, Jiayun; Hao, Guijie; Shen, Jinyu

    2016-01-01

    The giant freshwater prawn, Macrobrachium rosenbergii, is an economically important crustacean and is farmed in many countries. Since 2009, a larval mortality syndrome of M. rosenbergii has broken out and spread widely in the main breeding area, including Zhejiang, Jiangsu, Guangxi, and Guangdong Provinces in mainland China. A novel virus, named Macrobrachium rosenbergii Taihu virus (MrTV), was isolated from the moribund larvae and was determined to be the causative agent of the M. rosenbergii larval mortality syndrome by experimental infection. Further genomic sequencing suggested that the MrTV genome is monopartite, 10,303 nt in length, and dicistronic with two non-overlapping open reading frames (ORFs) separated by an intergenic region (IGR) and flanked by untranslated regions (UTRs). Phylogenetic analysis using the full-length genomic sequence and the putative amino acid sequences of the capsid protein revealed that MrTV was more closely related to the taura syndrome virus (TSV) than to any other viruses. According to these molecular features, we proposed that MrTV is a new species in the genus Aparavirus, family Dicistroviridae. These results may shed light on controlling larval mortality syndrome in M. rosenbergii. PMID:26848659

  14. First report on vertical transmission of a plasmid DNA in freshwater prawn, Macrobrachium rosenbergii.

    PubMed

    Chowdhury, Labrechai Mog; Gireesh-Babu, P; Pavan-Kumar, A; Suresh Babu, P P; Chaudhari, Aparna

    2014-09-01

    Outbreak of WSSV disease is one of the major stumbling blocks in shrimp aquaculture. DNA vaccines have shown potential for mass scale vaccination owing to their stability, cost effectiveness and easy maintenance. Development of economically feasible delivery strategies remains to be a major challenge. This study demonstrates vertical transmission of a plasmid DNA in a decapod Macrobrachium rosenbergii for the first time. Females at three different maturation stages (immature, matured and berried) and mature males were injected with a plasmid DNA and allowed to spawn with untreated counterparts. Using specific primers the plasmid DNA could be amplified from the offspring of all groups except that of berried females. For this confirmation genomic DNA was isolated from 3 pools of 10 post larvae in each group. This presents an ideal strategy to protect young ones at zero stress. PMID:24933022

  15. 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

  16. Effects of chlordecone on 20-hydroxyecdysone concentration and chitobiase activity in a decapod crustacean, Macrobrachium rosenbergii.

    PubMed

    Lafontaine, Anne; Gismondi, Eric; Boulangé-Lecomte, Céline; Geraudie, Perrine; Dodet, Nathalie; Caupos, Fanny; Lemoine, Soazig; Lagadic, Laurent; Thomé, Jean-Pierre; Forget-Leray, Joëlle

    2016-07-01

    Chlordecone (CLD) is an organochlorine insecticide abundant in aquatic environment of the French West Indies. However, few studies have investigated its impact on freshwater invertebrates. Whereas CLD is suspected of inducing endocrine disruption, this work aimed to study the effects of environmentally relevant concentrations of CLD on the 20-hydroxyecdysone (20-HE) hormone concentration and on the chitobiase activity, both having key roles in the molting process of crustaceans. In addition, the bioaccumulation of CLD was measured in the muscle tissue of Macrobrachium rosenbergii to underline potential dose-response relationship. The results have shown that CLD was bioaccumulated in exposed organisms according to a trend to a dose-response relationship. Moreover, it was observed that CLD decreased the 20-HE concentration in exposed prawns when compared to control, whatever the duration of exposure, as well as it inhibited the chitobiase activity after 30days of exposure. The present study indicates that CLD could interfere with molting process of M. rosenbergii by disturbing the 20-HE concentration and the activity of chitobiase, suggesting consequences at the long term on the shrimp development. This study also confirmed that CLD could be an endocrine disruptor in decapod crustaceans, as it was already observed in vertebrates. PMID:27108204

  17. 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 IκB-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

  18. 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.

  19. 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

  20. Quantification of lectin in freshwater prawn (Macrobrachium rosenbergii ) hemolymph by ELISA.

    PubMed

    Agundis, C; Pereyra, A; Zenteno, R; Brassart, C; Sierra, C; Vazquez, L; Zenteno, E

    2000-10-01

    An enzyme-linked immunoabsorbent assay was developed to quantify the lectin present in the hemolymph of the freshwater prawn Macrobrachium rosenbergii. This method involves the use of murine monoclonal IgG1 with kappa light chain (designated as 3G1) antibodies raised against the purified lectin, the assay that we developed recognized as little as 30 ng/ml of lectin, and was used to measure the lectin concentration in animals at different maturation stages. The highest concentration of lectin was identified in the hemolymph from post-larval prawns and the lowest in molt stage adult animals. The hemagglutination activity of the lectin was four-fold higher in adult than in juvenile specimens, although in all cases N-acetylated sugar residues, such as N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, and N-acetyl-D-neuraminic acid were inhibitors of the lectin activity, suggesting that lectin plays a role in the transport of N-acetylated sugar in juvenile prawns. Our results indicate that lectin concentration and hemagglutinating activity could be influenced by developmental conditions of the freshwater prawn. PMID:11079370

  1. Effects of Two Densities of Caged Monosex Nile Tilapia Oreochromis Niloticus on Water Quality, Phytoplankton Populations and Production When Polycultured With Macrobrachium rosenbergii in Temperate Ponds

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Elevated pH levels from the photosynthetic activity of phytoplankton can be a major problem in semi-intensive freshwater prawn (Macrobrachium rosenbergii) production systems. Phytoplanktontiverous fish may be able to graze excess phytoplankton and lower pH. The effect of different densities of cag...

  2. 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

  3. Genetic Inheritance of Female and Male Morphotypes in Giant Freshwater Prawn Macrobrachium rosenbergii

    PubMed Central

    2014-01-01

    Giant freshwater prawn (GFP) Macrobrachium rosenbergii is unique with males categorized in five different morphotypes (small claw, orange claw, blue claw, old blue claw and no claw males) and females in three reproductive statuses (mature ovary, berried and spawned females). In the present study we examined genetic inheritance of female and male morphotypes, their body weights and genetic associations between morphotypes and body traits. Restricted maximum likelihood fitting a multi-trait animal model was performed on a total of 21,459 body records collected over five generations in a GFP population selected for high growth rate. The estimates of variance components showed that there were substantial differences in additive genetic variance in body weight between male morphotypes. The low and significantly different from one genetic correlations between the expressions of body weight in male morphotypes also suggest that these traits should be treated as genetically different traits in selective breeding programs. By contrast, body weights of female types are essentially the same characters as indicated by the high genetic correlations between homologous trait expressions. In addition to body weight, male morphotypes and female reproductive statuses were treated as traits in themselves and were analysed as binary observations using animal and sire linear mixed models, and logit and probit threshold models. The estimates of heritability back-transformed from the liability scale were in good agreement with those obtained from linear mixed models, ranging from 0.02 to 0.43 for male morphotypes and 0.06 to 0.10 for female types. The genetic correlations among male morphoptypes were generally favourable. Body weight showed negative genetic associations with SM (−0.96), whereas those of body weight with other male morphotypes were positive (0.25 to 0.76). Our results showed that there is existence of heritable (additive genetic) component for male morphotypes, giving prospects for genetic selection to change population structure of GFP. PMID:24587246

  4. 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

  5. Gene silencing of VP9 gene impairs WSSV infectivity on Macrobrachium rosenbergii.

    PubMed

    Alenton, Rod Russel R; Kondo, Hidehiro; Hirono, Ikuo; Maningas, Mary Beth B

    2016-03-01

    White Spot Syndrome Virus (WSSV) remains the most widespread and devastating infectious agent that hit the shrimp aquaculture industry worldwide. To date, there are no known effective strategies yet to combat WSSV infection. Hence, functional studies on genes critical for viral infection is essential in elucidating shrimp-virus interaction. Here we report the function of a gene from WSSV coding for a non-structural protein, VP9, utilizing RNA interference. Silencing of VP9 gene also effectively suppressed other gene region in the WSSV genome (wsv168 gene) as early as day 1 post infection (dpi). Three set-ups using Macrobrachium rosenbergii shrimp were prepared for treatment using VP9-dsRNA, GFP-dsRNA, and PBS. Each shrimp was challenge with WSSV, and survival rate was recorded. VP9- and GFP-dsRNA injected shrimps showed a significant survival rate of 80% and 70%, respectively, in contrast to 0% of the PBS injected shrimps at 25dpi. Re-infection of shrimp survivors using a higher viral titer concentration, concurrent with the infection of new shrimp samples for the PBS control group, resulted in a significant 67% survival rate for VP9-dsRNA compared to 0% with that of GFP-dsRNA and PBS group. Challenge test on two more species, Penaeus monodon and Marsupenaeus japonicus, also significantly increased survival after VP9-dsRNA treatment. Our results provided evidence that VP9 gene plays an essential role in WSSV replication and it can be a potent target gene in the development of RNAi therapeutics for shrimps. PMID:26811904

  6. 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

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

    PubMed

    Suwansa-Ard, Saowaros; Thongbuakaew, Tipsuda; Wang, Tianfang; Zhao, Min; Elizur, Abigail; Hanna, Peter J; 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

  8. 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

  9. Identification and cloning of a transglutaminase from giant freshwater prawn, Macrobrachium rosenbergii, and its transcription during pathogen infection and moulting.

    PubMed

    Liu, Chun-Hung; Chang, Chin-Chyuan; Chiu, Yun-Chih; Cheng, Winton; Yeh, Maw-Sheng

    2011-12-01

    Complementary (c)DNA encoding transglutaminase (TG) messenger (m)RNA of the giant freshwater prawn, Macrobrachium rosenbergii, was cloned from haemocytes by a reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) using oligonucleotide primers based on the TG sequence of the horseshoe crab, Tachypleus tridentatus; tiger shrimp, Penaeus monodon; kuruma shrimp, Marsupenaeus japonicus; and crayfish, Pacifastacus leniusculus. The 2722-bp cDNA contained an open reading frame (ORF) of 2334 bp, a 72-bp 5'-untranslated region (UTR), and a 316-bp 3'-UTR containing a stop codon and a poly A tail. The molecular mass of the deduced amino acid (aa) sequence (778 aa) was 86.67 kDa with an estimated pI of 5.4. The M. rosenbergii TG (abbreviated MrTG, accession no.: JF309296) contains a typical transglutaminase-like homologue, two putative integrin-binding motifs (RGD), ten glycosylation sites, and four calcium-binding sites; a catalytic triad is present as in arthropod TGs. Sequence comparison and a phylogenetic analysis revealed that shrimp TG can be separated into three subgroups, penaeid TG1, freshwater crustacean TG2 and marine crustacean TG2, and MrTG was more closely related to TG2 than to TG1. MrTG mRNA and TG activities were detected in all tested tissues of M. rosenbergii, with MrTG mainly being synthesised by haemocytes. There was a negative correlation between clotting time of haemolymph, and MrTG expression and TG activity of haemocytes in prawn injected with Lactococcus garvieae. The pattern of MrTG mRNA expression and TG activity in haemocytes exhibited a contrary tendency with clotting time of haemolymph during the moult stages. Those results indicate that cloned MrTG is involved in the defence response, and is probably the major functional TG for haemolymph coagulation in M. rosenbergii. PMID:21854853

  10. Roles of dopamine receptors in mediating acute modulation of immunological responses in Macrobrachium rosenbergii.

    PubMed

    Chang, Zhong-Wen; Ke, Zhi-Han; Chang, Chin-Chyuan

    2016-02-01

    Dopamine (DA) was found to influence the immunological responses and resistance to pathogen infection in invertebrates. To clarify the possible modulation of DA through dopamine receptors (DAR) against acute environmental stress, the levels of DA, glucose and lactate in the haemolymph of Macrobrachium rosenbergii under hypo- and hyperthermal stresses were measured. The changes in immune parameters such as total haemocyte count (THC), differential haemocyte count (DHC), phenoloxidase (PO) activity, respiratory bursts (RBs), superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities and phagocytic activity (PA) were evaluated in prawns which received DAR antagonists (SCH23390, SCH, D1 antagonist; domperidone, DOM, D2 antagonist; chlorpromazine, CH, D1+2 antagonist) followed by hypo- (15 °C) and hyperthermal (34 °C) stresses. In addition, pharmacological analysis of the effect DA modulation was studied in haemocytes incubated with DA and DAR antagonists. The results revealed a significant increase in haemolymph DA accompanied with upregulated levels of glucose and lactate in prawns exposed to both hypo- and hyperthermal stresses in 2 h. In addition, a significant decrease in RBs per haemocyte was noted in prawns which received DAR antagonists when they exposed to hyperthermal stress for 30 min. In in vitro test, antagonism on RBs, SOD and GPx activity of haemocytes were further evidenced through D1, D1, D1+D2 DARs, respectively, in the meantime, no significant difference in PO activity and PA was observed among the treatment groups. These results suggest that the upregulation of DA, glucose and lactate in haemolymph might be the response to acute thermal stress for the demand of energy, and the DAR occupied by its antagonistic action impart no effect on immunological responses except RBs in vivo even though the modulation mediated through D1 DAR was further evidenced in RBs, SOD and GPx activities in vitro. It is therefore concluded that thermal stress mediate stress responses not only through DAR but also via diverse pathways, and DA might modulate the levels of RBs, SOD and GPx activities mainly through D1 DAR. PMID:26766178

  11. 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

  12. Isolation and characterization of a female-specific DNA marker in the giant freshwater prawn Macrobrachium rosenbergii

    PubMed Central

    Ventura, T; Aflalo, E D; Weil, S; Kashkush, K; Sagi, A

    2011-01-01

    In this study, a female-specific DNA marker in the freshwater prawn Macrobrachium rosenbergii was identified through amplified fragment length polymorphism (AFLP). The AFLP-derived sequence-characterized amplified region (SCAR) marker was tested in over 200 individuals, giving reproducible sex identification. Further molecular characterization of the sex-marker's genomic region (∼3 kb long) revealed the presence of tandem and inverted repeats. The ∼3-kb sequence was identified both in male and female prawns, but with subtle differences: a deletion of 3 bp (present in female prawn but absent in male prawn) identified upstream of the SCAR marker sequence and two female-specific single-nucleotide polymorphisms, both indicating that male prawns are homozygous, whereas female prawns are heterozygous in this locus. Fluorescent in situ hybridization showed the ∼3-kb sequence to be unique: to the best of our knowledge, this is the first report of a unique sex-specific sequence observed in situ in crustaceans. The sex-specific marker identified in M. rosenbergii may have considerable applied merit for crustacean culture in that it will enable the determination of genetic sex at early developmental stages when phenotypic differences are not identifiable. PMID:21522169

  13. 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

  14. 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 β-1,3-glucan-binding protein: LGBP, proPO, peroxinectin: PE, and α2-macroglobulin: α2-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 α2M 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

  15. Single CRD containing lectin from Macrobrachium rosenbergii (MrLec) participates in innate immunity against pathogen infections.

    PubMed

    Huang, Xin; Li, Wen; Jin, Min; Ma, Fu-Tong; Huang, Ying; Shi, Yan-Ru; Zhao, Ling-Ling; Feng, Jin-Ling; Ren, Qian; Wang, Wen

    2016-04-01

    As a type of pattern-recognition proteins, lectins perform important functions in the innate immunity of crustaceans, including prawns. Although several reports showed that C-type lectin domain family (CLEC) importantly functions in host-pathogen interactions, limited research has focused on CLEC in Macrobrachium rosenbergii. In the present study, a new single CRD containing CLEC (designated as MrLec) was reported in freshwater prawns, M. rosenbergii. The full-length cDNA of MrLec consisted of 1027 bp with an open reading frame of 801 bp, which encoded a peptide of 266 amino acid residues. Genomic sequence for MrLec was also obtained from the M. rosenbergii, which contain 4 exons and 3 introns. MrLec was found to contain a single carbohydrate-recognition domain with an EPN motif. MrLec was ubiquitously distributed in various tissues of a normal prawn, particularly in the hepatopancreas and gills. MrLec expression in the gills was significantly upregulated after a challenge with Vibrio parahaemolyticus and downregulated at 24 h after MrLec RNA interference (MrLec-RNAi). The expression levels of some AMPs, including antilipopolysaccharide factor 1 (Alf1) and lysozyme 2 (Lyso2), also markedly decreased after MrLec-RNAi. Recombinant MrLec can agglutinate (Ca(2+)-dependent) and bind both Gram-negative and Gram-positive bacteria. Results suggested that MrLec participates in the recognition of invading pathogens and functions in the immune response of prawn against pathogen infections. PMID:26923244

  16. Morphological and histological studies on freshwater prawn Macrobrachium rosenbergii (de man) irradiated with (60)Co gamma radiation.

    PubMed

    Stalin, A; Broos, K V; Sadiq Bukhari, A; Syed Mohamed, H E; Singhal, R K; Venu-Babu, P

    2013-11-15

    This study was framed to investigate the (60)Co gamma radiation induced morphological and histological variations in freshwater prawn Macrobrachium rosenbergii. The LD50 value of (60)Co gamma irradiated M. rosenbergii observed (by probit analysis) at 30 Gy. Prawns were irradiated to four different dose levels (3 mGy, 30 mGy, 300 mGy and 3,000 mGy) using Theratron Phoenix TeleCobalt Unit [P-33] and one control group (without irradiation) maintained separately. Irradiated groups exhibited several morphological variations such as discoloration; damaged rostrum; opaque coloration in cephalothorax; black bands and dot formation in abdomen; deformed uropods and telson in tail regions when compared with control group. The Hepato Somatic Index reflected the severity of radiation on hepatopancreas. Histological variations in gills, hepatopancreas and muscles of irradiated groups were observed. In gills, structural changes such as swollen and fused lamellae, abnormal gill tips, hyperplasic, necrotic and clavate-globate lamellae were observed in gamma irradiated prawns. Accumulation of hemocytes in hemocoelic space, interstitial sinuses filled with abnormal infiltrated hemocytes, the tubular epithelium with ruptured basal laminae, abnormal and coagulated lumen, necrotic tubules, thickened basal laminae, tissue debris, necrotic hepatocytes were observed in irradiated prawn hepatopancreas. In muscle, shrinkage of muscular fiber and necrotic musculature were observed in irradiated prawns. These structural alterations of the organs it is felt could affect the vital physiological functions such as respiration, osmotic and ionic regulation in gills and muscles; absorption, storage and secretion of the hepatopancreas which in turn could adversely affect the growth and survival of freshwater prawn M. rosenbergii. PMID:24140570

  17. A new nodavirus is associated with covert mortality disease of shrimp.

    PubMed

    Zhang, Qingli; Liu, Qun; Liu, Shuang; Yang, Haolin; Liu, Sun; Zhu, Luoluo; Yang, Bing; Jin, Jiting; Ding, Lixue; Wang, Xiuhua; Liang, Yan; Wang, Qintao; Huang, Jie

    2014-12-01

    A new nodavirus, named covert mortality nodavirus (CMNV), is associated with covert mortality disease of shrimp which has caused serious loss in China since 2009. Histopathological examination of shrimp suffering the disease revealed coagulative necrosis of striated muscle similar to typical histopathology features of infectious myonecrosis virus (IMNV), Penaeus vannamei nodavirus (PvNV) and Macrobrachium rosenbergii nodavirus (MrNV). However, shrimp suffering this disease tested negative for IMNV, MrNV and PvNV by reverse transcription (RT)-PCR. Additionally, eosinophilic inclusions were found in epithelium of the tubules in the hepatopancreas and lymphoid organ, and mass karyopyknotic nuclei existed in the muscle and lymphoid organ. The tubular epithelium of the hepatopancreas showed significant atrophy. A cDNA library was constructed from total RNA of infected shrimp. Sequencing and alignment analysis showed that one clone with an 1185 bp insert (designated CMNV-7) shared 54, 53 and 39% identity with the amino acid sequences of RNA-dependent RNA polymerase from Flock House virus, black beetle virus and MrNV. The results of fluorescence in situ hybridization showed that the hepatopancreas, striated muscle and lymphoid organ were positively reacting tissues. The mean size of negative-stained virus particles was 32 nm. In addition, a nested RT-PCR assay was developed for CMNV, and the RT-PCR detection results revealed that Fenneropenaeus chinensis, Litopenaeus vannamei and Marsupenaeus japonicus suffering from this disease were CMNV-positive. PMID:25205685

  18. Optimizing Hybrid de Novo Transcriptome Assembly and Extending Genomic Resources for Giant Freshwater Prawns (Macrobrachium rosenbergii): The Identification of Genes and Markers Associated with Reproduction.

    PubMed

    Jung, Hyungtaek; Yoon, Byung-Ha; Kim, Woo-Jin; Kim, Dong-Wook; Hurwood, David A; Lyons, Russell E; Salin, Krishna R; Kim, Heui-Soo; Baek, Ilseon; Chand, Vincent; Mather, Peter B

    2016-01-01

    The giant freshwater prawn, Macrobrachium rosenbergii, a sexually dimorphic decapod crustacean is currently the world's most economically important cultured freshwater crustacean species. Despite its economic importance, there is currently a lack of genomic resources available for this species, and this has limited exploration of the molecular mechanisms that control the M. rosenbergii sex-differentiation system more widely in freshwater prawns. Here, we present the first hybrid transcriptome from M. rosenbergii applying RNA-Seq technologies directed at identifying genes that have potential functional roles in reproductive-related traits. A total of 13,733,210 combined raw reads (1720 Mbp) were obtained from Ion-Torrent PGM and 454 FLX. Bioinformatic analyses based on three state-of-the-art assemblers, the CLC Genomic Workbench, Trans-ABySS, and Trinity, that use single and multiple k-mer methods respectively, were used to analyse the data. The influence of multiple k-mers on assembly performance was assessed to gain insight into transcriptome assembly from short reads. After optimisation, de novo assembly resulted in 44,407 contigs with a mean length of 437 bp, and the assembled transcripts were further functionally annotated to detect single nucleotide polymorphisms and simple sequence repeat motifs. Gene expression analysis was also used to compare expression patterns from ovary and testis tissue libraries to identify genes with potential roles in reproduction and sex differentiation. The large transcript set assembled here represents the most comprehensive set of transcriptomic resources ever developed for reproduction traits in M. rosenbergii, and the large number of genetic markers predicted should constitute an invaluable resource for future genetic research studies on M. rosenbergii and can be applied more widely on other freshwater prawn species in the genus Macrobrachium. PMID:27164098

  19. Optimizing Hybrid de Novo Transcriptome Assembly and Extending Genomic Resources for Giant Freshwater Prawns (Macrobrachium rosenbergii): The Identification of Genes and Markers Associated with Reproduction

    PubMed Central

    Jung, Hyungtaek; Yoon, Byung-Ha; Kim, Woo-Jin; Kim, Dong-Wook; Hurwood, David A.; Lyons, Russell E.; Salin, Krishna R.; Kim, Heui-Soo; Baek, Ilseon; Chand, Vincent; Mather, Peter B.

    2016-01-01

    The giant freshwater prawn, Macrobrachium rosenbergii, a sexually dimorphic decapod crustacean is currently the world’s most economically important cultured freshwater crustacean species. Despite its economic importance, there is currently a lack of genomic resources available for this species, and this has limited exploration of the molecular mechanisms that control the M. rosenbergii sex-differentiation system more widely in freshwater prawns. Here, we present the first hybrid transcriptome from M. rosenbergii applying RNA-Seq technologies directed at identifying genes that have potential functional roles in reproductive-related traits. A total of 13,733,210 combined raw reads (1720 Mbp) were obtained from Ion-Torrent PGM and 454 FLX. Bioinformatic analyses based on three state-of-the-art assemblers, the CLC Genomic Workbench, Trans-ABySS, and Trinity, that use single and multiple k-mer methods respectively, were used to analyse the data. The influence of multiple k-mers on assembly performance was assessed to gain insight into transcriptome assembly from short reads. After optimisation, de novo assembly resulted in 44,407 contigs with a mean length of 437 bp, and the assembled transcripts were further functionally annotated to detect single nucleotide polymorphisms and simple sequence repeat motifs. Gene expression analysis was also used to compare expression patterns from ovary and testis tissue libraries to identify genes with potential roles in reproduction and sex differentiation. The large transcript set assembled here represents the most comprehensive set of transcriptomic resources ever developed for reproduction traits in M. rosenbergii, and the large number of genetic markers predicted should constitute an invaluable resource for future genetic research studies on M. rosenbergii and can be applied more widely on other freshwater prawn species in the genus Macrobrachium. PMID:27164098

  20. Identification and characterization of differentially expressed transcripts in the gills of freshwater prawn (Macrobrachium rosenbergii) under salt stress.

    PubMed

    Barman, Hirak Kumar; Patra, Swagat Kumar; Das, Varsha; Mohapatra, Shibani Dutta; Jayasankar, Pallipuram; Mohapatra, Chinmayee; Mohanta, Ramya; Panda, Rudra Prasanna; Rath, Surya Narayan

    2012-01-01

    The giant freshwater prawn, Macrobrachium rosenbergii, is an economically important species. It is a euryhaline shrimp, surviving in wide-range salinity conditions. A change in gene expression has been suggested as an important component for stress management. To better understand the osmoregulatory mechanisms mediated by the gill, a subtractive and suppressive hybridization (SSH) tool was used to identify expressed transcripts linked to adaptations in saline water. A total of 117 transcripts represented potentially expressed under salinity conditions. BLAST analysis identified 22% as known genes, 9% as uncharacterized showing homologous to unannotated ESTs, and 69% as unknown sequences. All the identified known genes representing broad spectrum of biological pathways were particularly linked to stress tolerance including salinity tolerance. Expression analysis of 10 known genes and 7 unknown/uncharacterized genes suggested their upregulation in the gills of prawn exposed to saline water as compared to control indicating that these are likely to be associated with salinity acclimation. Rapid amplification of cDNA ends (RACE) was used for obtaining full-length cDNA of MRSW-40 clone that was highly upregulated during salt exposure. The sequenced ESTs presented here will have potential implications for future understanding about salinity acclimation and/or tolerance of the prawn. PMID:22619594

  1. 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

  2. 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 μg·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

  3. Function of a novel C-type lectin with two CRD domains from Macrobrachium rosenbergii in innate immunity.

    PubMed

    Huang, Xin; Huang, Ying; Shi, Yan-Ru; Ren, Qian; Wang, Wen

    2015-03-01

    C-type lectins play crucial roles in innate immunity. In the present study, a novel C-type lectin gene, designated as MrCTL, was identified from Macrobrachium rosenbergii. MrCTL contains 2 carbohydrate-recognition domains (CRDs), namely MrCRD1 and MrCRD2. The MrCRD1 contains a QEP motif and MrCRD2 contains a motif of EPD. MrCTL was mainly expressed in the hepatopancreas. The expression level of MrCTL in hepatopancreas was significantly upregulated after a challenge with Vibrio parahaemolyticus or White spot syndrome virus (WSSV). The recombinant MrCTL, MrCRD1 and MrCRD2 have an ability to agglutinate both Gram-negative (V. parahaemolyticus) and Gram-positive bacteria (Staphylococcus aureus) in a calcium dependent manner. The recombinant MrCTL, MrCRD1 and MrCRD2 bind directly to all tested microorganisms. All these results suggested that MrCTL may have important roles in immune defense against invading pathogens in prawns. PMID:25475962

  4. Identification and Characterization of Differentially Expressed Transcripts in the Gills of Freshwater Prawn (Macrobrachium rosenbergii) under Salt Stress

    PubMed Central

    Barman, Hirak Kumar; Patra, Swagat Kumar; Das, Varsha; Mohapatra, Shibani Dutta; Jayasankar, Pallipuram; Mohapatra, Chinmayee; Mohanta, Ramya; Panda, Rudra Prasanna; Rath, Surya Narayan

    2012-01-01

    The giant freshwater prawn, Macrobrachium rosenbergii, is an economically important species. It is a euryhaline shrimp, surviving in wide-range salinity conditions. A change in gene expression has been suggested as an important component for stress management. To better understand the osmoregulatory mechanisms mediated by the gill, a subtractive and suppressive hybridization (SSH) tool was used to identify expressed transcripts linked to adaptations in saline water. A total of 117 transcripts represented potentially expressed under salinity conditions. BLAST analysis identified 22% as known genes, 9% as uncharacterized showing homologous to unannotated ESTs, and 69% as unknown sequences. All the identified known genes representing broad spectrum of biological pathways were particularly linked to stress tolerance including salinity tolerance. Expression analysis of 10 known genes and 7 unknown/uncharacterized genes suggested their upregulation in the gills of prawn exposed to saline water as compared to control indicating that these are likely to be associated with salinity acclimation. Rapid amplification of cDNA ends (RACE) was used for obtaining full-length cDNA of MRSW-40 clone that was highly upregulated during salt exposure. The sequenced ESTs presented here will have potential implications for future understanding about salinity acclimation and/or tolerance of the prawn. PMID:22619594

  5. Assessment of Constructed Wetland in Nutrient Reduction, in the Commercial Scale Experiment Ponds of Freshwater Prawn Macrobrachium rosenbergii.

    PubMed

    Mahmood, Tariq; Zhang, Jing; Zhang, Guosen

    2016-03-01

    A free water surface constructed wetland (CW) was integrated into two commercial ponds of Macrobrachium rosenbergii, to evaluate the role of CW in reducing the excess nutrient concentration and other pollutants produced from the aquaculture waste. Hydraulic residence time was kept constant (24 h). There was a significant (p < 0.05) decrease in total suspended solids (TSS, 73.2 ± 15.4 %) and total nitrogen (TN, 39.6 ± 44.2 %) between wetland inflow and wetland outflow. The performance of the CW was highly impacted by the low concentration of dissolved nutrients at the inflow of CW. Results showed about 43.8 ± 24.6 % NO3 (-), 25.7 ± 23.0 % NH4 (+), 14.3 ± 1.0 % NO2 (-), 28.4 ± 18.8 % DIN and 13.1 ± 10.0 % PO4 (3-) were removed. In agreement with previous published investigations, comparing values of pollutants before and after recirculation, this study concludes that a CW system can provide good water quality and minimize external water input. PMID:26679323

  6. Nutrients budget and effluents characteristics in polyculture of scampi (Macrobrachium rosenbergii) and Indian major carps ponds using organic inputs.

    PubMed

    Adhikari, Subhendu; Sahu, Bharat Chandra; Dey, Lambodar

    2012-01-01

    Experiments were conducted for the study of nutrient budget in farmers' ponds (0.4-0.6 ha) in Orissa, India, at stocking densities of 0.30-0.38/m(2) for scampi (Macrobrachium rosenbergii) and 0.60-0.70/m(2) for Indian major carps (Catla catla, Labeo rohita and Cirrhinus mrigala). The average initial body weights of scampi and the major carps were 0.02 and 8-10 g, respectively. The culture period was 272-292 days. Feed was the main input. The FCR (feed conversion ratio) varied from 1.78 to 1.83. Feed and cow dung were applied to these ponds as organic inputs. At harvest time, the average weight of scampi and carps varied from 73 to 92 g and from 718 to 820 g, respectively. Among all the inputs, feed alone accounted for 97.60% total nitrogen (N), 97.90% total phosphorus (P) and 94.72% total organic carbon (OC), respectively. The harvest of scampi and carps accounted for recovery of 52.45% N, 19.43% P and 18.12% OC, respectively. N, P and OC accumulated in sediment were 38.31, 71.40 and 69.50%, respectively. The median nutrient loads in the fish pond effluents were equivalent to 0.58 kg of inorganic N, 0.135 kg of P and 8.83 kg of total OC per ton of Indian major carps and scampi production. PMID:22864442

  7. Expression Profile of Antioxidant Enzymes in Hemocytes from Freshwater Prawn Macrobrachium rosenbergii Exposed to an Elevated Level of Copper.

    PubMed

    Guo, Hui; Miao, Yu-Tao; Xian, Jian-An; Qian, Kun; Wang, An-Li

    2015-10-01

    This study evaluated the expression level of antioxidant enzymes Cu, Zn-superoxide dismutase (Cu, Zn-SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione S-transferase (GST) in hemocytes of Macrobrachium rosenbergii exposed to copper by real-time PCR (qRT-PCR). Results showed that the mRNA expression of Cu, Zn-SOD increased to reach a peak at 6 h, then recovered to its normal level at 48 h. CAT expression level was significantly increased at 12 h and reached a peak at 24 h, but recovered to its normal level later. GPx expression level was significantly increased at 6 h and reached the peak at 12 h. GST expression level was significantly induced from 12 to 24 h and then dropped to its normal level at 48 h. These results indicated that antioxidant enzymes were inducible, possibly for removing excessive reactive oxygen species to protect prawn from oxidative stress. PMID:26215459

  8. Characterization of two novel ADP ribosylation factors from giant freshwater prawn Macrobrachium rosenbergii and their responses to WSSV challenge.

    PubMed

    Ding, Zheng-Feng; Ren, Jie; Tan, Jing-Min; Wang, Zheng; Yin, Shao-Wu; Huang, Ying; Huang, Xin; Wang, Wen; Lan, Jiang-Feng; Ren, Qian

    2015-01-01

    ADP-ribosylation factors (Arfs) are small GTP-binding proteins that have an essential function in intracellular trafficking and organelle structure. To date, little information is available on the Arfs in the economically important giant freshwater prawn Macrobrachium rosenbergii and their relationship to viral infection. Here we identified two Arf genes from M. rosenbergii (MrArf1 and MrArf2) for the first time. Phylogenetic analysis showed that MrArf1, together with MjArf1 from shrimp Marsupenaeus japonicus belonged to Class I Arfs. By contrast, MrArf2 didn't not match any of the Arfs classes of I/II/III, although it could be clustered with an Arf protein from M. japonicas called MjArfn, which may represent an analog of the Arf. MrArf1 was ubiquitously expressed in all the examined tissues, with the highest transcription level in the hepatopancreas, whereas MrArf2 was only highly expressed in the hepatopancreas and exhibited very low levels in the heart, stomach, gills and intestine. The expression level of MrArf1 in the gills was down-regulated post 24 h WSSV challenge, and reached the maximal level at 48 h. MrArf1 in the hepatopancreas went up from 24 to 48 h WSSV challenge. MrArf2 transcript in the gill also went down at 24 h and then was upregulated at 48 h WSSV challenge. MrArf2 increased significantly in the hepatopancreas 24 h after infection and then went down at 48 h WSSV challenge. RNAi results showed that knockdown of MrArf1 or MrArf2 could inhibit the expression of the envelope protein gene vp28 of the WSSV. So, it could be speculated that MrArf1 and MrArf2 might play important roles in the innate immune system against WSSV infection. PMID:25451300

  9. Characterization of a gC1qR from the giant freshwater prawn, Macrobrachium rosenbergii.

    PubMed

    Ye, Ting; Huang, Xin; Wang, Xian-Wei; Shi, Yan-Ru; Hui, Kai-Min; Ren, Qian

    2015-03-01

    gC1qR, as a multicompartmental and a multifunctional protein, plays an important role in innate immunity. In this study, a gC1qR homolog (MrgC1qR) in the giant freshwater prawn, Macrobrachium rosenbergii was identified. MrgC1qR, a 258-amino-acid polypeptide, shares high identities with gC1qR from other species. MrgC1qR gene was expressed in different tissues and was highest expressed in the hepatopancreas. In addition, the MrgC1qR transcript was significantly enhanced after 6 h of white spot syndrome virus (WSSV) infection or post 2 h, 24 h of Vibrio anguillarum challenge compared to appropriate controls. Moreover, recombinant MrgC1qR (rMrgC1qR) had bacterial binding activity, the result also revealed that rMrgC1qR could bind pathogen-associated molecular patterns (PAMPs) such as LPS or PGN, suggesting that MrgC1qRmight function as a pathogen-recognition receptor (PRR). Furthermore, glutathione S-transferase (GST) pull-down assays showed that rMrgC1qR with GST-tag could bind to rMrFicolin1 or rMrFicolin2 with His-tag. Altogether, these results may demonstrate a role for MrgC1qR in innate immunity in the giant freshwater prawns. PMID:25555810

  10. Gene profiling and characterization of arginine kinase-1 (MrAK-1) from freshwater giant prawn (Macrobrachium rosenbergii).

    PubMed

    Arockiaraj, Jesu; Vanaraja, Puganeshwaran; Easwvaran, Sarasvathi; Singh, Arun; Alinejaid, Tahereh; Othman, Rofina Yasmin; Bhassu, Subha

    2011-07-01

    Arginine kinase-1 (MrAK-1) was sequenced from the freshwater prawn Macrobrachium rosenbergii using Illumina Solexa Genome Analyzer Technique. MrAK-1 consisted of 1068 bp nucleotide encoded 355 polypeptide with an estimated molecular mass of 40 kDa. MrAK-1 sequence contains a potential ATP:guanido phosphotransferases active domain site. The deduced amino acid sequence of MrAK-1 was compared with other 7 homologous arginine kinase (AK) and showed the highest identity (96%) with AK-1 from cherry shrimp Neocaridina denticulate. The qRT-PCR analysis revealed a broad expression of MrAK-1 with the highest expression in the muscle and the lowest in the eyestalk. The expression of MrAK-1 after challenge with the infectious hypodermal and hematopoietic necrosis virus (IHHNV) was tested in muscle. In addition, MrAK-1 was expressed in Escherichia coli by prokaryotic expression plasmid pMAL-c2x. The optimum temperature (30 °C) and pH (8.5) was determined for the enzyme activity assay. MrAK-1 showed significant (P < 0.05) activity towards 10-50 mM ATP concentration. The enzyme activity was inhibited by α-ketoglutarate, glucose and ATP at the concentration of 10, 50 and 100 mM respectively. Conclusively, the findings of this study indicated that MrAK-1 might play an important role in the coupling of energy production and utilization and the immune response in shrimps. PMID:21549198

  11. Effects of dietary administration of water hyacinth (Eichhornia crassipes) extracts on the immune responses and disease resistance of giant freshwater prawn, Macrobrachium rosenbergii.

    PubMed

    Chang, Chin-Chyuan; Tan, Hui-Ching; Cheng, Winton

    2013-07-01

    The hot-water extract of Eichhornia crassipes leaves (ECE) was produced and incorporated into the diet of the prawn, Macrobrachium rosenbergii, as an immunostimulant. Survival rates of prawn against Lactococcus garvieae, and its immune parameters including the total haemocyte count (THC), different haemocyte count (DHC), phenoloxidase (PO) activity, respiratory bursts (RBs), superoxide dismutase (SOD) activity, glutathione peroxidase (GPx) activity, transglutaminase (TG) activity, haemolymph coagulation time, and phagocytic activity and clearance efficiency against L. garvieae by M. rosenbergii were determined when prawn (23.0 ± 2.8 g) were fed ECE-containing diets at 0, 1.0, 2.0 and 3.0 g kg(-1). Prawn fed a diet containing ECE at 2.0 and 3.0 g kg(-1) for 12 days showed significantly increased THC, HC, GC, PO activity, RBs, SOD activity, GPx activity, and TG activity, and a significantly decreased coagulation time. The phagocytic activity and clearance efficiency against L. garvieae of prawn fed the ECE-containing diets at 1.0, 2.0, and 3.0 g kg(-1) were significantly higher than those of prawn fed the control diet at 3-12 days. Survival rates of M. rosenbergii fed the diet containing ECE at concentrations of 2 and 3 g kg(-1) were significantly higher than those fed the control diet after challenge with L. garvieae for 48-144 h. The relative percentage survival of prawn fed the 1.0, 2.0, and 3.0 g kg(-1) ECE-containing diets for 12 days were 17.5%, 39.1%, and 52.2%. It was concluded that the ECE can be used as an immunostimulant for prawn through dietary administration to enhance immune responses and resistance of M. rosenbergii against L. garvieae. PMID:23603238

  12. 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

  13. Ultrastructural changes during spermatogenesis, biochemical and hormonal evidences of testicular toxicity caused by TBT in freshwater prawn Macrobrachium rosenbergii (De Man, 1879).

    PubMed

    Revathi, Peranandam; Iyapparaj, Palanisamy; Vasanthi, Lourduraj Arockia; Munuswamy, Natesan; Krishnan, Muthukalingan

    2014-10-01

    The present investigation documents the impact of tributyltin (TBT) on the ultrastructural variation of spermatogenesis in freshwater prawn Macrobrachium rosenbergii. The environmentally realistic concentration of TBT can cause damages to the endocrine and reproductive physiology of crustaceans. In this context, three concentrations viz. 10, 100, and 1000 ng/L were selected and exposed to prawns for 90 days. The TBT exposed prawn exhibited decrease the reproductive activity as evidenced by sperm count and sperm length compared to control. Histopathological results revealed the retarded testicular development, abnormal structure of seminiferous tubule, decrease in the concentration of spermatozoa, diminution of seminiferous tubule membrane, abundance of spermatocytes and vacuolation in testis of treated prawns. Ultrastructural study also confirmed the impairment of spermatogenesis in treated prawns. Furthermore, radioimmunoassay (RIA) clearly documented the reduction of testosterone level in TBT exposed groups. Thus, TBT substantially reduced the level of male sex hormone as well as biochemical constituents which ultimately led to impairment of spermatogenesis in the freshwater male prawn M. rosenbergii. PMID:23362226

  14. The effect of sugars and free amino acids from the freshwater prawn Macrobrachium rosenbergii hemolymph on lectin activity and on oxidative burst.

    PubMed

    Soria, Frida; Sierra, Claudia; Bouquelet, Stephane; Brassart, Colette; Agundis, Concepción; Zenteno, Edgar; Vázquez, Lorena

    2006-01-01

    We determined the effect of low molecular weight components (LMWC) from healthy juvenile and adult Macrobrachium rosenbergii hemolymph on lectin activity and oxidative burst (OB) in hemocytes. In an attempt to identify the LMWC that affect the lectin's hemagglutinating activity or oxidative burst, we determined the hemolymph carbohydrates and free amino acids (FAA) concentration. The LMWC (<2000 Da) were obtained after dialysis of the hemolymph. Our results showed that LMWC from juveniles exerted a greater inhibition on lectin than LMWC from adult hemolymph. Production of superoxide radicals by hemocytes was lower in the presence of juvenile (p<0.05) as compared to adult LMWC. FAA composition of the hemolymph and of LMWC from adults showed higher proportion of alanine (which corresponded to 25% of total FAA) and proline (>20%); whereas, in juveniles, the main FAA identified were glycine (>40%) and alanine (26%). N-Acetyl-D-glucosamine (GlcNAc) was the main sugar residue in the hemolymph and LMWC from juveniles; its concentration was 2.4 times higher than glucose (Glc), whereas, in adults, Glc was the main free sugar residue. Our results suggest that the proportion of FAA and carbohydrates in the hemolymph of M. rosenbergii seems to be correlated with the maturation process; furthermore, the high proportion of free GlcNAc and glycine regulate, in the juvenile stage, lectin activity and cellular oxidative mechanisms, respectively. PMID:16290085

  15. 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

  16. Isolated and combined exposure to ammonia and nitrite in giant freshwater pawn (Macrobrachium rosenbergii): effects on the oxidative stress, antioxidant enzymatic activities and apoptosis in haemocytes.

    PubMed

    Zhang, Yufan; Ye, Chaoxia; Wang, Anli; Zhu, Xuan; Chen, Changhong; Xian, Jianan; Sun, Zhenzhu

    2015-10-01

    The residual contaminators such as ammonia and nitrite are widely considered as relevant sources of aquatic environmental pollutants, posing a great threat to shrimp survival. To study the toxicological effects of ammonia and nitrite exposure on the innate immune response in invertebrates, we investigated the oxidative stress and apoptosis in haemocytes of freshwater prawn (Macrobrachium rosenbergii) under isolated and combined exposure to ammonia and nitrite in order to provide useful information about adult prawn immune responses. M. rosenbergii (13.44 ± 2.75 g) were exposed to 0, 5, and 25 mg/L total ammonia-N (TAN) and 0, 5, and 20 mg/L nitrite-N for 24 h. All ammonia concentrations were combined with all nitrite concentrations, making a total of nine treatments studied. Following the exposure treatment, antioxidant enzyme activity, reactive oxygen species (ROS) generation, nitric oxide (NO) generation, and apoptotic cell ratio of haemocytes were measured using flow cytometry. Results indicated that ROS generation was sensitive to the combined effect of ammonia and nitrite, which subsequently affected the Cu-Zn SOD activity. In addition, CAT showed the highest activity at 5 mg/L TAN while GPx decreased at 5 mg/L TAN and returned towards baseline at 25 mg/L. NO generation synchronized with the apoptotic cell ratio in haemocytes, indicating that NO production was closely associated with programmed cell death. Both NO production and apoptotic ratios significantly decreased following 25 mg/L TAN, which may be due to the antagonistic regulation of NO and GPx. We hypothesized that the toxicological effect of nitrite exhibited less change in physiological changes compared to that of ammonia, because of the high tolerance to nitrite exposure in mature M. rosenbergii and/or the competitive effects of chloride ions. Taken together, these results showed that ammonia and nitrite caused a series of combined oxidative stress and apoptosis in M. rosenbergi, but further studies are of great need to explain the mechanisms. PMID:25967939

  17. Simultaneous identification and quantification of 4-cumylphenol, 2,4-bis-(dimethylbenzyl)phenol and bisphenol A in prawn Macrobrachium rosenbergii.

    PubMed

    Zuo, Yuegang; Zhu, Zhuo

    2014-07-01

    Bisphenol A (BPA), 4-cumylphenol (4-CP) and 2,4-bis-(dimethylbenzyl)phenol (2,4-DCP) are all high production volume chemicals and widely used in plastic and other consumer products. During the past two decades, BPA has attracted a great deal of scientific and public attention due to its presence in the environment and estrogenic property. Although 4-CP and 2,4-DCP are much more estrogenic and toxic than BPA, little information is available about their occurrence and fate in the environment. In this study, a rapid, selective, accurate and reliable analytical method was developed for the simultaneous determination of 4-CP, 2,4-DCP and BPA in prawn Macrobrachium rosenbergii. The method comprises an ultrasound-accelerated extraction followed by capillary gas chromatographic (GC) separation. The detection limits range from 1.50 to 36.4 ng kg(-1) for the three alkylphenols. The calibration curves are linear over the concentration range tested with the coefficients of determination, R(2), greater than 0.994. The developed method was successfully applied to the simultaneous determination of 4-CP, 2,4-DCP and BPA in prawn samples. The peak identification was confirmed using GC-MS. Bisphenol A, 2,4-bis-(dimethylbenzyl)phenol and 4-cumylphenol were found in prawn samples in the concentration ranges of 0.67-5.51, 0.36-1.61, and 0.00-1.96 ng g(-1) (wet weight), respectively. All relative standard deviations are less than 4.8%. At these environmentally relevant concentration levels, 4-CP, 2,4-DCP and BPA may affect the reproduction and development of aquatic organisms, including negative influence on crustaceans' larval survival, molting, metamorphosis and shell hardening. This is the first study reported on the occurrence of 4-CP, 2,4-DCP and BPA in prawn M. rosenbergii. PMID:24560775

  18. 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.

  19. Dietary supplementation of green synthesized manganese-oxide nanoparticles and its effect on growth performance, muscle composition and digestive enzyme activities of the giant freshwater prawn Macrobrachium rosenbergii.

    PubMed

    Asaikkutti, Annamalai; Bhavan, Periyakali Saravana; Vimala, Karuppaiya; Karthik, Madhayan; Cheruparambath, Praseeja

    2016-05-01

    The green synthesized Mn3O4 nanoparticles (manganese-oxide nanoparticles) using Ananas comosus (L.) peel extract was characterized by various techniques. HR-SEM photograph showed that manganese-oxide nanoparticles (Mn-oxide NPs) were spherical in shape, with an average size of 40-50nm. The Zeta potential revealed the surface charge of Mn-oxide NPs to be negative. Further, the Mn-oxide NPs were dietary supplemented for freshwater prawn Macrobrachium rosenbergii. The experimental basal diets were supplemented with Mn-oxide NPs at the rates of 0 (control), 3.0, 6.0, 9.0, 12, 15 and 18mg/kg dry feed weight. The as-supplemented Mn-oxide NPs were fed in M. rosenbergii for a period of 90 days. The experimental study demonstrated that prawns fed with diet supplemented with 3-18mg Mn-oxide NPs/kg shows enhanced (P<0.05) growth performance, including final weight and weight gain (WG). Significant differences (P<0.05) in feed conversion ratio (FCR) were observed in prawn fed with different diets. Additionally, prawns fed with 3.0-18mg/kg Mn-oxide NPs supplemented diets achieved significant (P<0.05) improvement in growth performance, digestive enzyme activities and muscle biochemical compositions, while, the prawns fed with 16mg/kg of Mn-oxide NPs showed enhanced performance. Prawns fed on diet supplemented with 16mg/kg Mn-oxide NPs showed significantly (P<0.05) higher total protein level. The antioxidants enzymatic activity (SOD and CAT) metabolic enzymes status in muscle and hepatopancreas showed no significant (P>0.05) alterations in prawns fed with 3.0-18mg/kg of Mn-oxide NPs supplemented diets. Consequently, the present work proposed that 16mg/kg of Mn-oxide NPs could be supplemented for flexible enhanced survival, growth and production of M. rosenbergii. Therefore, the data of the present study recommend the addition of 16mg/kg of Mn-oxide NPs diet to developed prawn growth and antioxidant defense system. PMID:27049122

  20. 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

  1. Cells producing insulin-like androgenic gland hormone of the giant freshwater prawn, Macrobrachium rosenbergii, proliferate following bilateral eyestalk-ablation.

    PubMed

    Phoungpetchara, Ittipon; Tinikul, Yotsawan; Poljaroen, Jaruwan; Chotwiwatthanakun, Charoonroj; Vanichviriyakit, Rapeepun; Sroyraya, Morakot; Hanna, Peter J; Sobhon, Prasert

    2011-06-01

    We found that the androgenic gland (AG) of Macrobrachium rosenbergii possesses three cell types. Type I cells are small polygonal shaped-cells (13.4 μm in diameter), stain strongly with hematoxylin-eosin (H&E), have abundant multilayered rough endoplasmic reticulum (rER), and nuclei containing mostly heterochromatin. Type II cells are slightly larger (18.6 μm in diameter), stain lightly with H&E, have rER with dilated cisternae, and nuclei containing mostly euchromatin. Type III cells (previously undescribed) are similar in size and shape to type I cells, but the cytoplasm is unstained and they have a high amount of smooth endoplasmic reticulum (sER) and mitochondria with tubular cristae. Bilateral eyestalk-ablation resulted in AG hypertrophy with a proliferation and predominance of type I cells as determined by bromodeoxyuridine (BrdU) assays. Expression of insulin-like androgenic gland hormone (Mr-IAG), determined by immunohistochemistry, was weak in type I cells, strong in type II cells of both the intact and eyestalk-ablated, and negative in type III cells. It was also detected in spermatogonia, nurse cells, and epithelium lining of the spermatic duct. The function of Mr-IAG in these tissues is yet to be elucidated but the distribution implies a strong role in male reproduction. PMID:21477831

  2. 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

  3. 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.

  4. 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

  5. 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

  6. 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 β-1,3-glucan-binding protein, LGBP; prophenoloxidase, proPO; peroxinectin, PE; α2-macroglobulin, α2-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 β-AR antagonist of metoprolol (Met), on the SOD activity by the β-AR antagonist of propranolol (Pro), on the RB by the β-AR antagonist of Met and prazosin (Pra), and on the proPO system-related genes expressions by α-AR antagonist of Pra. These results show that NE modulates prawn haemocytes proPO system-related genes expressions via α1-AR, PO activity and phagocytosis via β1-AR, respiratory burst via α1-and β1-ARs, and SOD activity via β2-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 α1-, β1-and β2-ARs. PMID:26210701

  7. Responses of three very large inducible GTPases to bacterial and white spot syndrome virus challenges in the giant fresh water prawn Macrobrachium rosenbergii.

    PubMed

    Huang, Ying; Jin, Min; Yin, Shaowu; Ding, Zhengfeng; Wang, Wen; Ren, Qian

    2016-04-01

    Interferons (IFNs) are cytokines secreted by cells in response to invasion by pathogens, such as viruses, bacteria, parasites, or tumor cells. Very large inducible GTPases (VLIG) are the latest IFN-inducible GTPase family to be discovered and are the largest known GTPases of any species. However, VLIG proteins from invertebrates have yet to be characterized. In this study, three forms of VLIGs designated as MrVLIG1, MrVLIG2, and MrVLIG3 were cloned from the giant fresh water prawn Macrobrachium rosenbergii. MrVLIG1 has a 5445 bp open reading frame (ORF) encoding an 1814-amino acid protein. The complete nucleotide sequence of MrVLIG2 cDNA is 7055 bp long consisting of a 5757 bp ORF encoding a protein with 1918 amino acids. The full length of the MrVLIG3 gene consists of 5511 bp with a 3909 bp ORF encoding a peptide with 1302 amino acids. BLASTP and phylogenetic tree analyses showed that the three MrVLIGs are clustered into one subgroup and, together with other vertebrate VLIGs, into a branch. Tissue distribution analysis indicated that the mRNAs of the three MrVLIGs were widely expressed in almost all detected tissues, including the hemocytes, heart, hepatopancreas, gills, stomach, and intestine, with the highest expression in the hepatopancreas. They were also detected in the intestine but with relatively low expression levels. Quantitative real-time RT-PCR analysis showed that the mRNA transcripts of the MrVLIGs in the hepatopancreas were significantly expressed at various time points after infection with Vibrio parahaemolyticus and white spot syndrome virus. In summary, the three isoforms of VLIG genes participate in the innate immune response of the shrimps to bacterial and viral infections. PMID:26850335

  8. 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.

  9. Ultrastructural and sequence characterization of Penaeus vannamei nodavirus (PvNV) from Belize.

    PubMed

    Tang, Kathy F J; Pantoja, Carlos R; Redman, Rita M; Navarro, Solangel A; Lightner, Donald V

    2011-05-01

    The Penaeus vannamei nodavirus (PvNV), which causes muscle necrosis in Penaeus vannamei from Belize, was identified in 2005. Infected shrimp show clinical signs of white, opaque lesions in the tail muscle. Under transmission electron microscopy, the infected cells exhibit increases in various organelles, including mitochondria, Golgi stacks, and rough endoplasmic reticulum. Cytoplasmic inclusions containing para-crystalline arrays of virions were visualized. The viral particle is spherical in shape and 19 to 27 nm in diameter. A cDNA library was constructed from total RNA extracted from infected shrimp. Through nucleotide sequencing from the cDNA clones and northern blot hybridization, the PvNV genome was shown to consist of 2 segments: RNA1 (3111 bp) and RNA2 (1183 bp). RNA1 contains 2 overlapped open reading frames (ORF A and B), which may encode a RNA-dependent RNA polymerase (RdRp) and a B2 protein, respectively. RNA2 contains a single ORF that may encode the viral capsid protein. Sequence analyses showed the presence of 4 RdRp characteristic motifs and 2 conserved domains (RNA-binding B2 protein and viral coat protein) in the PvNV genome. Phylogenetic analysis based on the translated amino acid sequence of the RdRp reveals that PvNV is a member of the genus Alphanodavirus and closely related to Macrobrachium rosenbergii nodavirus (MrNV). In a study investigating potential PvNV vectors, we monitored the presence of PvNV by RT-PCR in seabird feces and various aquatic organisms collected around a shrimp farm in Belize. PvNV was detected in mosquitofish, seabird feces, barnacles, and zooplankton, suggesting that PvNV can be spread via these carriers. PMID:21790065

  10. 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

  11. Development of in situ hybridization and RT-PCR assay for the detection of a nodavirus (PvNV) that causes muscle necrosis in Penaeus vannamei.

    PubMed

    Tang, Kathy F J; Pantoja, Carlos R; Redman, Rita M; Lightner, Donald V

    2007-05-01

    A nodavirus (tentatively named PvNV, Penaeus vannamei nodavirus) that causes muscle necrosis in P. vannamei was found in Belize in 2004. From 2004 to 2006, shrimp samples collected from Belize exhibited clinical signs, white, opaque lesions in the tails and histopathology similar to those of shrimps infected by infectious myonecrosis virus (IMNV). Histological examination revealed multifocal necrosis and hemocytic fibrosis in the skeletal muscle. In addition, basophilic, cytoplasmic inclusions were found in striated muscle, lymphoid organ and connective tissues. However, IMNV was not detected in these shrimps by either RT-PCR or in situ hybridization, suggesting that these lesions may be caused by another RNA virus. Thus, a cDNA library was constructed from total RNA extracted from hemolymph collected from infected shrimp. One clone (designated PvNV-4) with a 928 bp insert was sequenced and found to be similar (69% similarity when comparing the translated amino acid sequences) to the capsid protein gene of MrNV (Macrobrachium rosenbergii nodavirus). The insert of PvNV-4 was labeled with digoxigenin-11-deoxyuridine triphosphate (dUTP) and hybridized to tissue sections of P. vannamei with muscle necrosis collected in Belize and from laboratory bioassays. The samples were positive for PvNV infection. Positively reacting tissues included skeletal muscle, connective tissues, the lymphoid organ, and hemocytes in the heart and gills. In addition, we experimentally infected both P. vannamei and P. monodon with PvNV prepared from Belize samples. A nested RT-PCR assay developed from the PvNV-4 cloned sequence showed that both species are susceptible to PvNV infection. PMID:17629112

  12. 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

  13. Macrobrachium rosenbergii mannose binding lectin: synthesis of MrMBL-N20 and MrMBL-C16 peptides and their antimicrobial characterization, bioinformatics and relative gene expression analysis.

    PubMed

    Arockiaraj, Jesu; Chaurasia, Mukesh Kumar; Kumaresan, Venkatesh; Palanisamy, Rajesh; Harikrishnan, Ramasamy; Pasupuleti, Mukesh; Kasi, Marimuthu

    2015-04-01

    Mannose-binding lectin (MBL), an antimicrobial protein, is an important component of innate immune system which recognizes repetitive sugar groups on the surface of bacteria and viruses leading to activation of the complement system. In this study, we reported a complete molecular characterization of cDNA encoded for MBL from freshwater prawn Macrobrachium rosenbergii (Mr). Two short peptides (MrMBL-N20: (20)AWNTYDYMKREHSLVKPYQG(39) and MrMBL-C16: (307)GGLFYVKHKEQQRKRF(322)) were synthesized from the MrMBL polypeptide. The purity of the MrMBL-N20 (89%) and MrMBL-C16 (93%) peptides were confirmed by MS analysis (MALDI-ToF). The purified peptides were used for further antimicrobial characterization including minimum inhibitory concentration (MIC) assay, kinetics of bactericidal efficiency and analysis of hemolytic capacity. The peptides exhibited antimicrobial activity towards all the Gram-negative bacteria taken for analysis, whereas they showed the activity towards only a few selected Gram-positive bacteria. MrMBL-C16 peptides produced the highest inhibition towards both the Gram-negative and Gram-positive bacteria compared to the MrMBL-N20. Both peptides do not produce any inhibition against Bacillus sps. The kinetics of bactericidal efficiency showed that the peptides drastically reduced the number of surviving bacterial colonies after 24 h incubation. The results of hemolytic activity showed that both peptides produced strong activity at higher concentration. However, MrMBL-C16 peptide produced the highest activity compared to the MrMBL-N20 peptide. Overall, the results indicated that the peptides can be used as bactericidal agents. The MrMBL protein sequence was characterized using various bioinformatics tools including phylogenetic analysis and structure prediction. We also reported the MrMBL gene expression pattern upon viral and bacterial infection in M. rosenbergii gills. It could be concluded that the prawn MBL may be one of the important molecule which is involved in antimicrobial mechanism. Moreover, MrMBL derived MrMBL-N20 and MrMBL-C16 peptides are important antimicrobial peptides for the recognition and eradication of viral and bacterial pathogens. PMID:25575476

  14. Induction of humoral and cell-mediated immune responses by hepatitis B virus epitope displayed on the virus-like particles of prawn nodavirus.

    PubMed

    Yong, Chean Yeah; Yeap, Swee Keong; Goh, Zee Hong; Ho, Kok Lian; Omar, Abdul Rahman; Tan, Wen Siang

    2015-02-01

    Hepatitis B virus (HBV) is a deadly pathogen that has killed countless people worldwide. Saccharomyces cerevisiae-derived HBV vaccines based upon hepatitis B surface antigen (HBsAg) is highly effective. However, the emergence of vaccine escape mutants due to mutations on the HBsAg and polymerase genes has produced a continuous need for the development of new HBV vaccines. In this study, the "a" determinant within HBsAg was displayed on the recombinant capsid protein of Macrobrachium rosenbergii nodavirus (MrNV), which can be purified easily in a single step through immobilized metal affinity chromatography (IMAC). The purified protein self-assembled into virus-like particles (VLPs) when observed under a transmission electron microscope (TEM). Immunization of BALB/c mice with this chimeric protein induced specific antibodies against the "a" determinant. In addition, it induced significantly more natural killer and cytotoxic T cells, as well as an increase in interferon gamma (IFN-γ) secretion, which are vital for virus clearance. Collectively, these findings demonstrated that the MrNV capsid protein is a potential carrier for the HBV "a" determinant, which can be further extended to display other foreign epitopes. This paper is the first to report the application of MrNV VLPs as a novel platform to display foreign epitopes. PMID:25416760

  15. 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 β-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

  16. 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

  17. 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 μg/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.

  18. 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...

  19. Two Kazal-type protease inhibitors from Macrobrachium nipponense and Eriocheir sinensis: comparative analysis of structure and activities.

    PubMed

    Qian, Ye-Qing; Li, Ye; Yang, Fan; Yu, Yan-Qin; Yang, Jin-Shu; Yang, Wei-Jun

    2012-03-01

    Kazal-type inhibitors (KPIs) play important roles in many biological and physiological processes, such as blood clotting, the immune response and reproduction. In the present study, two male reproductive tract KPIs, termed Man-KPI and Ers-KPI, were identified in Macrobrachium nipponense and Eriocheir sinensis, respectively. The inhibitory activities of recombinant Man-KPI and Ers-KPI against chymotrypsin, elastase, trypsin and thrombin were determined. The results showed that both of them strongly inhibit chymotrypsin and elastase. Kinetic studies were performed to elucidate their inhibition mechanism. Furthermore, individual domains were also expressed to learn further which domain contributes to the inhibitory activities of intact KPIs. Only Man-KPI_domain3 is active in the inhibition of chymotrypsin and elastase. Meanwhile, Ers-KPI_domain2 and 3 are responsible for inhibition of chymotrypsin, and Ers-KPI_domains2, 3 and 4 are responsible for the inhibition of elastase. Meanwhile, the inhibitory activities of these two KPIs toward Macrobrachium rosenbergii, M. nipponense and E. sinensis sperm were compared with that of the Kazal-type peptidase inhibitor (MRPINK) characterized from the M. rosenbergii reproductive tract in a previous study. The results demonstrated that KPIs can completely inhibit the gelatinolytic activities of sperm proteases from their own species, while different levels of cross-inhibition were observed between KPI and proteases from different species. These results may provide new perspective to further clarify the mechanism of KPI-proteases interaction in the male reproductive system. PMID:22200638

  20. Isolation and characterization of a nodavirus associated with mass mortality in Asian seabass (Lates calcarifer) from the west coast of India.

    PubMed

    Banerjee, Debashis; Hamod, Mohammed A; Suresh, Thangavel; Karunasagar, Indrani

    2014-12-01

    A fish nodavirus was detected in the juveniles of Asian seabass (Lates calcarifer) during a massive outbreak in the seabass cage culture farm located in the south west coast of India. The clinical signs of the disease included anorexia, inflated abdomen, exophthalmia, darkening of the whole body, erratic swimming and cork-screw type movement followed by death. The dead and the moribund fish were analyzed for nodavirus by reverse transcriptase-polymerase chain reaction (RT-PCR) using specific primers targeting the T4 region of RNA2 coat protein gene. This is the first report of nodavirus infection in the fresh water cage-reared seabass fish in the west coast of India. The piscine nodavirus was detected in the brain, retina and kidney of all the fishes examined. The PCR products were cloned and sequenced. The sequence analysis showed more than 90 % homology with the other coat protein gene sequence of piscine nodaviruses from other countries. The phylogenetic analysis based on the partial nucleotide sequence of RNA2 coat protein gene revealed that the virus belonged to the red-spotted grouper nervous necrosis virus, which is one of the widely distributed genotype among the other four known genotypes of piscine nodavirus. PMID:25674617

  1. Identification of five anti-lipopolysaccharide factors in oriental river prawn, Macrobrachium nipponense.

    PubMed

    Wang, Yili; Tang, Ting; Gu, Jihai; Li, Xiang; Yang, Xue; Gao, Xiaobin; Liu, Fengsong; Wang, Jianhui

    2015-10-01

    Anti-lipopolysaccharide factors (ALFs) are a group of antimicrobial peptides (AMPs) with broad-spectrum antimicrobial activities and antiviral activities mainly found in crustaceans and horseshoe crabs. In the present study, we identified 5 ALF expression sequence tags (ESTs) through analysis of the established M. nipponense transcriptome, and cloned their full-length cDNA sequences using rapid amplification of cDNA ends (RACE) method. The 5 ALFs were designated as MnALF1-5, and all of them showed high similarity with their Macrobrachium rosenbergii homologs in the phylogenetic analyses, especially in LPS binding domains. In healthy adult prawns, we found the highest expression of MnALF2 and MnALF4 in haemocytes, and the highest expression of MnALF4 and MnALF3 in intestine. Some isoforms of MnALF were down-regulated but the majority was up-regulated in different prawn tissues upon Aeromonas hydrophila challenge. To conform the expected antimicrobial activities harbored in MnALFs' LPS binding domains, we used a synthesized peptide cMnALF24 that corresponds to the LPS binding domain of MnALF2 as a representative molecule for the antibacterial activity test, and found that cMnALF24 possessed strong and broad-spectrum antibacterial activity against Gram positive and Gram negative bacteria, but no inhibition activity against fungi; Meanwhile, in the hemolytic test, cMnALF24 showed weak hemolysis activities (around 10%) to the rabbit red blood cells at concentrations of 0.67-33.50 μM. This study provides insights into understanding the antibacterial function of ALFs in the innate immunity of freshwater prawn, and reports a peptide that can be a potential drug candidate with good efficacy against bacterial infection and low toxicity to host cells. PMID:26159095

  2. 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

  3. Atlantic halibut experimentally infected with nodavirus shows increased levels of T-cell marker and IFNγ transcripts.

    PubMed

    Overgård, Aina-Cathrine; Nerland, Audun Helge; Fiksdal, Ingrid Uglenes; Patel, Sonal

    2012-05-01

    The transcript levels of viral RNAs, selected T-cell marker and cytokine genes, toll like receptor (TLR) 7, and two interferon stimulated genes (ISG) were analysed in sexually immature adult Atlantic halibut (Hippoglossus hippoglossus L.) experimentally infected with nodavirus. The expression of the T-cell markers, TLR7 and the cytokine genes was further explored in in vitro stimulated anterior kidney leucocytes (AK leucocytes) isolated from the experiment fish and from additional untreated non-injected fish. The levels of viral RNA1 and RNA2 were increasing in brain and eye at around 4 and 8weeks post injection (wpi), respectively, and still increasing at the end of the experiment, especially in eye. Immuno-positive cells and signs of vacuolisation in both brain and eye were seen at 14wpi. Increased transcript levels of TCRβ, CD4-2, CD4, CD8α, and Lck in brain and eye of the experimentally infected halibut suggested an involvement of halibut T-cells in the immune response against nodavirus. Interestingly, a similar expression pattern of TCRβ, CD4 and Lck was seen in both brain and eye. However, compared to brain that showed elevated transcript levels of TCRβ, CD4 and Lck mainly at 10 and 14wpi, the increase appeared earlier between 3 and 4wpi in the eye. Yet, an increase in the transcript level of IFNγ was seen at 10 and 14wpi in both organs. Moreover, elevated levels of TLR7, IL-1β, IL-6, ISG15 and Mx were detected in vivo. The in vitro experiments, stimulating AK leucocytes with ConA-PMA, imiquimod or nodavirus, further supported an involvement of IL-6 and IFNγ in the immune response against nodavirus and the involvement of CD8β(+) cells. Results from the present study thus indicate an importance of T-cells, IFNγ and the analysed ISGs in the immune response against nodavirus in Atlantic halibut, and would be of great help in future vaccination trials giving the possibility to monitor the immune response rather than mortality during post-vaccination challenge experiments. PMID:22020051

  4. Genome-Wide Analysis of Host Factors in Nodavirus RNA Replication

    PubMed Central

    Hao, Linhui; Lindenbach, Brett; Wang, Xiaofeng; Dye, Billy; Kushner, David; He, Qiuling; Newton, Michael; Ahlquist, Paul

    2014-01-01

    Flock House virus (FHV), the best studied of the animal nodaviruses, has been used as a model for positive-strand RNA virus research. As one approach to identify host genes that affect FHV RNA replication, we performed a genome-wide analysis using a yeast single gene deletion library and a modified, reporter gene-expressing FHV derivative. A total of 4,491 yeast deletion mutants were tested for their ability to support FHV replication. Candidates for host genes modulating FHV replication were selected based on the initial genome-wide reporter gene assay and validated in repeated Northern blot assays for their ability to support wild type FHV RNA1 replication. Overall, 65 deletion strains were confirmed to show significant changes in the replication of both FHV genomic RNA1 and sub-genomic RNA3 with a false discovery rate of 5%. Among them, eight genes support FHV replication, since their deletion significantly reduced viral RNA accumulation, while 57 genes limit FHV replication, since their deletion increased FHV RNA accumulation. Of the gene products implicated in affecting FHV replication, three are localized to mitochondria, where FHV RNA replication occurs, 16 normally reside in the nucleus and may have indirect roles in FHV replication, and the remaining 46 are in the cytoplasm, with functions enriched in translation, RNA processing and trafficking. PMID:24752411

  5. 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

  6. 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

  7. Cell Culture Isolation of Piscine Nodavirus (Betanodavirus) in Fish-Rearing Seawater.

    PubMed

    Nishi, Shinnosuke; Yamashita, Hirofumi; Kawato, Yasuhiko; Nakai, Toshihiro

    2016-04-15

    Piscine nodavirus (betanodavirus) is the causative agent of viral nervous necrosis (VNN) in a variety of cultured fish species, particularly marine fish. In the present study, we developed a sensitive method for cell culture isolation of the virus from seawater and applied the method to a spontaneous fish-rearing environment. The virus in seawater was concentrated by an iron-based flocculation method and subjected to isolation with E-11 cells. A real-time reverse transcriptase PCR (RT-PCR) assay was used to quantify the virus in water. After spiking into seawater was performed, a betanodavirus strain (redspotted grouper nervous necrosis virus [RGNNV] genotype) was effectively recovered in the E-11 cells at a detection limit of approximately 10(5)copies (equivalent to 10(2)50% tissue culture infective doses [TCID50])/liter seawater. In an experimental infection of juvenile sevenband grouper (Epinephelus septemfasciatus) with the virus, the virus was isolated from the drainage of a fish-rearing tank when the virus level in water was at least approximately 10(5)copies/liter. The application of this method to sevenband grouper-rearing floating net pens, where VNN prevailed, resulted in the successful isolation of the virus from seawater. No differences were found in the partial sequences of the coat protein gene (RNA2) between the clinical virus isolates of dead fish and the cell-cultured virus isolates from seawater, and the viruses were identified as RGNNV. The infection experiment showed that the virus isolates from seawater were virulent to sevenband grouper. These results showed direct evidence of the horizontal transmission of betanodavirus via rearing water in marine aquaculture. PMID:26896128

  8. 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

  9. 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

  10. Antimicrobial response is increased in the testis of European sea bass, but not in gilthead seabream, upon nodavirus infection.

    PubMed

    Valero, Yulema; García-Alcázar, Alicia; Esteban, M Ángeles; Cuesta, Alberto; Chaves-Pozo, Elena

    2015-05-01

    Antimicrobial peptides (AMPs) have a crucial role in the fish innate immune response, being considered a fundamental component of the first line of defence against pathogens. Moreover, AMPs have not been studied in the fish gonad since this is used by some pathogens as a vehicle or a reservoir to be transmitted to the progeny, as occurs with nodavirus (VNNV), which shows vertical transmission through the gonad and/or gonadal fluids, but no study has looked into the gonad of infected fish. In this framework, we have characterized the antimicrobial response triggered by VNNV in the testis of European sea bass, a very susceptible species of the virus, and in the gilthead seabream, which acts as a reservoir, both in vivo and in vitro, and compared with that present in the serum and brain (target tissue of VNNV). First, our data show a great antiviral response in the brain of gilthead seabream and in the gonad of European sea bass. In addition, for the first time, our results demonstrate that the antimicrobial activities (complement, lysozyme and bactericidal) and the expression of AMP genes such as complement factor 3 (c3), lysozyme (lyz), hepcidin (hamp), dicentracin (dic), piscidin (pis) or β-defensin (bdef) in the gonad of both species are very different, but generally activated in the European sea bass, probably related with the differences of susceptibility upon VNNV infection, and even differs to the brain response. Furthermore, the in vitro data suggest that some AMPs are locally regulated playing a local immune response in the gonad, while others are more dependent of the systemic immune system. Data are discussed in the light to ascertain their potential role in viral clearance by the gonad to avoid vertical transmission. PMID:25707600

  11. 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 μg (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 μg (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 μg (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 μg (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

  12. RNA Binding by a Novel Helical Fold of B2 Protein from Wuhan Nodavirus Mediates the Suppression of RNA Interference and Promotes B2 Dimerization ▿

    PubMed Central

    Qi, Nan; Cai, Dawei; Qiu, Yang; Xie, Jiazheng; Wang, Zhaowei; Si, Jie; Zhang, Jiamin; Zhou, Xi; Hu, Yuanyang

    2011-01-01

    Wuhan nodavirus (WhNV) is a newly identified member of the Nodaviridae family with a bipartite genome of positive-sense RNAs. A nonstructural protein encoded by subgenomic RNA3 of nodaviruses, B2, serves as a potent RNA silencing suppressor (RSS) by sequestering RNA duplexes. We have previously demonstrated that WhNV B2 blocks RNA silencing in cultured Drosophila cells. However, the molecular mechanism by which WhNV B2 functions remains unknown. Here, we successfully established an RNA silencing system in cells derived from Pieris rapae, a natural host of WhNV, by introducing into these cells double-stranded RNA (dsRNA)-expressing plasmids or chemically synthesized small interfering RNAs (siRNAs). Using this system, we revealed that the WhNV B2 protein inhibited Dicer-mediated dsRNA cleavage and the incorporation of siRNA into the RNA-induced silencing complex (RISC) by sequestering dsRNA and siRNA. Based on the modeled B2 3-dimensional structure, serial single alanine replacement mutations and N-terminal deletion analyses showed that the RNA-binding domain of B2 is formed by its helices α2 and α3, while helix α1 mediates B2 dimerization. Furthermore, positive feedback between RNA binding and B2 dimerization was uncovered by gel shift assay and far-Western blotting, revealing that B2 dimerization is required for its binding to RNA, whereas RNA binding to B2 in turn promotes its dimerization. All together, our findings uncovered a novel RNA-binding mode of WhNV B2 and provided evidence that the promotion effect of RNA binding on dimerization exists in a viral RSS protein. PMID:21734038

  13. 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

  14. A new species of Macrobrachium Spence Bate, 1868 (Decapoda, Palaemonidae), M. pantanalense, from the Pantanal, Brazil.

    PubMed

    Dos Santos, Antonina; Hayd, Liliam; Anger, Klaus

    2013-01-01

    The neotropical species Macrobrachium amazonicum (Heller, 1862) is considered a particularly successful species, showing an extremely wide range of distribution (ca 4.000 km across). Populations assigned to this species live in estuaries along the northern and northeastern coasts of South America as well as in fresh water habitats in the Amazon, Orinoco and Paraná-Paraguay river basins. Following recent comparative studies that showed differential ecological, reproductive, developmental and physiological traits in geographically separated populations from the Amazon delta and the Pantanal region, Brazil, we examined the morphology of adult shrimps from these two regions. Based on significant differences, we conclude that the Pantanal population constitutes a new species, which is described here as Macrobrachium pantanalense. The main differences between M. amazonicum and the new species have been found in the morphology of the second pereiopod, the telson, and in the color patterns of both males and females. A modification on the key of American species of Macrobrachium is provided to accommodate the new species. PMID:26106741

  15. Macrobrachium indianum (Decapoda: Palaemonidae), a new species of hill stream prawn from Pambar River, Kerala, India.

    PubMed

    Pillai, P M; Unnikrishnan, V; Prasannan, K

    2015-01-01

    Macrobrachium indianum new species is described from the Pambar River, Kerala, S. India. The species shares certain characters with M. gurudeve Jayachandran & Raji, 2004, M. bombayense Almelker & Sankolli, 2006 and M. kulkarnii Almelker & Sankolli, 2006, while it differs remarkably from these three species in distinctive diagnostic characters: rostral formula 7-8/3-4 with 1 postorbital teeth, one tooth above orbit; carapace smooth with distal end of rostrum directed downwards; cephalothorax longer than rostrum; in second chelate leg, proximal cutting edge of movable finger with two weak denticles, one weak denticle in immovable finger, carpus longer than merus, merus shorter than propodus and longer than ischium; dactylus the shortest podomere. Five thick and a few thin reddish brown bands of chromatophores are seen on carapace. Pigmentation is found mid and ventro-laterally on abdominal segments, pereiopods have chromatophores at the distal part of podomeres. PMID:25947453

  16. Complete larval development of the Monkey River Prawn Macrobrachium lar (Palaemonidae) using a novel greenwater technique.

    PubMed

    Lal, Monal M; Seeto, Johnson; Pickering, Timothy D

    2014-01-01

    This study documents the complete larval development of the Monkey River Prawn Macrobrachium lar using a new greenwater rearing technique. Approximately 6,000 larvae were reared for 110 days at an initial stocking density of 1 ind./6 L. Salinity at hatch was 10 ± 2 ppt and progressively increased to 30 ± 2 ppt until decapodids had metamorphosed. Temperature was maintained at 28 ± 0.5°C, pH at 7.8 ± 0.2, DO2 > 6.5 mg/L and NH(4+) and NH3 ≤ 1.5 and ≤0.1 ppm respectively throughout the culture period. Larval development was extended and occurred through 13 zoeal stages, with the first decapodid measuring 6.2 ± 0.63 mm in total length observed after 77 days. 5 decapodids in total were produced, and overall survival to this stage was 0.08%. Overall, the pattern of larval growth shares similarities with those of other Macrobrachium spp. that have a prolonged/normal type of development, and it is likely that larvae underwent mark time moulting which contributed to the lengthened development duration. While this study represents a significant breakthrough in efforts to domesticate M. lar, improvement of larval survival rates and decreased time till metamorphosis are required before it can become fully viable for commercial scale aquaculture. PMID:25332868

  17. Bioaccumulation and retention kinetics of cadmium in the freshwater decapod Macrobrachium australiense.

    PubMed

    Cresswell, Tom; Simpson, Stuart L; Smith, Ross E W; Nugegoda, Dayanthi; Mazumder, Debashish; Twining, John

    2014-03-01

    The potential sources and mechanisms of cadmium bioaccumulation by the native freshwater decapods Macrobrachium species in the waters of the highly turbid Strickland River in Papua New Guinea were examined using (109)Cd-labelled water and food sources and the Australian species Macrobrachium australiense as a surrogate. Synthetic river water was spiked with environmentally relevant concentrations of cadmium and animals were exposed for 7 days with daily renewal of test solutions. Dietary assimilation of cadmium was assessed through pulse-chase experiments where prawns were fed separately (109)Cd-labelled fine sediment, filamentous algae and carrion (represented by cephalothorax tissue of water-exposed prawns). M. australiense readily accumulated cadmium from the dissolved phase and the uptake rate increased linearly with increasing exposure concentration. A cadmium uptake rate constant of 0.10 0.05 L/g/d was determined in synthetic river water. During depuration following exposure to dissolved cadmium, efflux rates were low (0.9 5%/d) and were not dependent on exposure concentration. Assimilation efficiencies of dietary sources were comparable for sediment and algae (48-51%), but lower for carrion (28 5%) and efflux rates were low (0.2-2.6%/d) demonstrating that cadmium was well retained by M. australiense. A biokinetic model of cadmium accumulation by M. australiense predicted that for exposures to environmentally relevant cadmium concentrations in the Strickland River, uptake from ingestion of fine sediment and carrion would be the predominant sources of cadmium to the organism. The model predicted the total dietary route would represent 70-80% of bioaccumulated cadmium. PMID:24508761

  18. Development and functional morphology of the mouthparts and foregut in larvae and post-larvae of Macrobrachium jelskii (Decapoda: Palaemonidae).

    PubMed

    Rocha, Cristina Pantoja; Souza, Adelson Silva de; Maciel, Murilo; Maciel, Cristiana R; Abrunhosa, Fernando Araújo

    2016-05-01

    The morphology of the mouthparts and foregut of the larvae and post-larvae of Macrobrachium jelskii was investigated to determine their functional roles in feeding, in order to understand the larval feeding behaviour and the changes that occur during its development. The mouthparts and foregut of the zoea I and II are morphologically similar, rudimentary and non-functional in feeding. Only in the final larval stage, zoea III, do the external mouthparts and foregut become structurally more complex and thus likely to play a potential role in feeding. Two behavioral trials (point of no return, point of reserve saturation) evaluated the resistance to starvation in zoea I, II, and III. The results indicate that they have sufficient nutritional reserves to permit them to complete metamorphosis without feeding. Overall, our results suggest that the zoea I and II of Macrobrachium jelskii engage in obligate lecithotrophy and zoea III in facultative lecithotrophy. PMID:26899315

  19. A molecular approach towards the taxonomy of fresh water prawns Macrobrachium striatum and M. equidens (Decapoda, Palaemonidae) using mitochondrial markers.

    PubMed

    Jose, Deepak; Nidhin, B; Anil Kumar, K P; Pradeep, P J; Harikrishnan, M

    2016-07-01

    Genus Macrobrachium includes freshwater prawns which inhabit most diverse habitats ranging from low saline areas to inland hill streams and impounded water bodies. Being morphologically conserved, this genus has been exposed to severe disputes related to their taxonomy, systematics and phylogeny. Macrobrachium striatum and M. equidens represent two morphologically related congeneric species within this genus. Earlier, M. striatum was considered as a striped form of M. equidens. Though these species are now well-described morphologically and differentiated into two species, no molecular level investigation has been carried out in support of their speciation. We report a study on M. striatum and M. equidens with emphasis to their molecular data through mitochondrial markers (16S ribosomal RNA and cytochrome oxidase subunit I). Results obtained from developed molecular markers of the two species revealed considerable genetic differentiation between them. Phylogram generated using Minimum evolution and Neighbour joining analyses differentiated M. striatum and M. equidens as two independent species. Genetic distance data showed high interspecific divergence (ranging from 3.9% to 17.0% for 16S rRNA sequences and 13.8% to 21.0% for COI sequences) between M. striatum and M. equidens confirming the findings of phylogram. Hence, it could be delineated that M. striatum and M. equidens represent two distinct species within genus Macrobrachium with emphasis to their morphology and genetics. PMID:26119113

  20. Bioaccumulation kinetics and organ distribution of cadmium and zinc in the freshwater decapod crustacean Macrobrachium australiense.

    PubMed

    Cresswell, Tom; Simpson, Stuart L; Mazumder, Debashish; Callaghan, Paul D; Nguyen, An P

    2015-01-20

    This study used the radioisotopes (109)Cd and (65)Zn to explore the uptake, retention and organ distribution of these nonessential and essential metals from solution by the freshwater decapod crustacean Macrobrachium australiense. Three treatments consisting of cadmium alone, zinc alone, and a mixture of cadmium and zinc were used to determine the differences in uptake and efflux rates of each metal individually and in the metal mixture over a three-week period, followed by depuration for 2 weeks in metal-free water using live-animal gamma-spectrometry. Following exposure, prawns were cryosectioned and the spatial distribution of radionuclides visualized using autoradiography. Metal uptake and efflux rates were the same in the individual and mixed-metal exposures, and efflux rates were close to zero. The majority of cadmium uptake was localized within the gills and hepatopancreas, while zinc accumulated in the antennal gland at concentrations orders of magnitude greater than in other organs. This suggested that M. australiense may process zinc much faster than cadmium by internally transporting the accumulated zinc to the antennal gland. The combination of uptake studies and autoradiography greatly increases our understanding of how metal transport kinetics and internal processing may influence the toxicity of essential and nonessential metals in the environment. PMID:25537180

  1. Characterization of the male-specific lethal 3 gene in the oriental river prawn, Macrobrachium nipponense.

    PubMed

    Zhang, Y P; Sun, S M; Fu, H T; Ge, X P; Qiao, H; Zhang, W Y; Xiong, Y W; Jiang, S F; Gong, Y S; Jin, S B

    2015-01-01

    In this study, male-specific lethal 3 homolog (Mnmsl3) was cloned and characterized from the freshwater prawn Macrobrachium nipponense (Crustacea: Decapoda: Palaemonidae) by rapid amplification of cDNA ends. The deduced amino acid sequences of Mnmsl3 showed high-sequence homology to the insect Msl3 and contained a conserved chromatin organization modifier domain and an MORF4-related gene domain. Real-time quantitative reverse transcription-polymerase chain reaction showed that the Mnmsl3 gene was expressed in all the investigated tissues, with the highest level of expression in the testis. The expression level of Mnmsl3 between males and females was different in the gonad (testis or ovary), abdominal ganglion, and heart. The results revealed that the Mnmsl3 gene might play roles in regulating chromatin and in dosage compensation of M. nipponense. Real-time quantitative reverse transcription-polymerase chain reaction also revealed that Mnmsl3 mRNA expression was significantly increased in both 5 and 20 days post-larvae after metamorphosis, suggesting that Mnmsl3 plays complex and important roles in the early embryonic development and sex differentiation of M. nipponense. PMID:25966075

  2. Assessment of acute toxicity of carbofuran in Macrobrachium olfersii (Wiegmann, 1836) at different temperature levels.

    PubMed

    Barbieri, Edison; Moreira, Priscila; Luchini, Luiz Alberto; Hidalgo, Karla Ruiz; Muñoz, Alejandro

    2016-01-01

    Carbofuran (2,3-dihydro-2,2-dimethyl-7-benzofuranyl methylcarbamate; C12H15NO3) is one of the most toxic carbamate pesticides. For acute toxicity of carbofuran, juveniles of Macrobrachium olfersii were exposed to different concentrations of carbofuran using the static renewal method at different temperature levels (15, 20 and 25°C) at pH 7.0. The main purpose of the present study was to detect the acute toxicity of carbofuran to M. olfersii and investigate its effects on oxygen consumption and ammonium excretion; these tests have not been carried out in this species before. First, the acute toxicity - median lethal concentration - of carbofuran to M. olfersii for 24, 48, 72 and 96 h was examined, which resulted in the following values: 1.64, 1.22, 0.86 and 0.42 mg L(-1), respectively. Furthermore, we also found that carbofuran caused an inhibition in oxygen consumption of 60.6, 65.3 and 66.2% with respect to the control. In addition, after separate exposures to carbofuran, elevations in ammonium excretion were more than 500% with respect to the control. PMID:23847016

  3. A novel C-type lectin is involved in the innate immunity of Macrobrachium nipponense.

    PubMed

    Xiu, Yunji; Wang, Yinghui; Bi, Jingxiu; Liu, Yuhan; Ning, Mingxiao; Liu, Hui; Li, Shuang; Gu, Wei; Wang, Wen; Meng, Qingguo

    2016-03-01

    C-type lectins (CTLs) play important roles in invertebrate innate immunity by recognizing and eliminating pathogens. In the present study, a low-density lipoprotein receptor class A (LDLa) domain-containing CTL was identified from the oriental river prawn Macrobrachium nipponense, designated as MnCTLDcp1. The full-length cDNA of MnCTLDcp1 was composed of 1462 bp, with a 999-bp ORF encoding a 332-aa protein. An LDLa and a single C-type lectin-like domain (CTLD) were found. The mRNA transcripts of MnCTLDcp1 was expressed the highest in heart. After the prawns were challenged by Aeromonas hydrophila and Staphylococcus aureus, the expression level of MnCTLDcp1 in heart and hemocytes were all significantly up-regulated. Sugar binding assay revealed that the MnCTLDcp1 could bind to the glycoconjugates of bacteria surface, such as LPS, PGN and they can compete with bacterial as competitors. The recombinant MnCTLDcp1 agglutinates Gram-positive (S. aureus and Bacillus subtilis) and Gram-negative bacteria (A. hydrophila, Vibrio parahaemolyticus, Escherichia coli and Pseudomonas aeruginosa) in the presence of calcium and also could bind to these bacteria. These results clearly suggested that MnCTLDcp1 functions as a pattern-recognition receptor involved in the innate immunity of M. nipponense. PMID:26804648

  4. Shrimp laminin receptor binds with capsid proteins of two additional shrimp RNA viruses YHV and IMNV.

    PubMed

    Busayarat, Nattaphon; Senapin, Saengchan; Tonganunt, Moltira; Phiwsaiya, Kornsunee; Meemetta, Watcharachai; Unajak, Sasimanas; Jitrapakdee, Sarawut; Lo, Chu-Fang; Phongdara, Amornrat

    2011-07-01

    Laminin receptor (Lamr) in shrimp was previously proposed to be a potential receptor protein for Taura syndrome virus (TSV) based on yeast two-hybrid assays. Since shrimp Lamr bound to the VP1 capsid protein of TSV, we were interested to know whether capsid/envelope proteins from other shrimp viruses would also bind to Lamr. Thus, capsid/envelope encoding genes from 5 additional shrimp viruses were examined. These were Penaeus stylirostris densovirus (PstDNV), white spot syndrome virus (WSSV), infectious myonecrosis virus (IMNV), Macrobrachium rosenbergii nodavirus (MrNV), and yellow head virus (YHV). Protein interaction analysis using yeast two-hybrid assay revealed that Lamr specifically interacted with capsid/envelope proteins of RNA viruses IMNV and YHV but not MrNV and not with the capsid/envelope proteins of DNA viruses PstDNV and WSSV. In vitro pull-down assay also confirmed the interaction between Lamr and YHV gp116 envelope protein, and injection of recombinant Lamr (rLamr) protein produced in yeast cells protected shrimp against YHV in laboratory challenge tests. PMID:21414409

  5. Immunological-based assays for specific detection of shrimp viruses.

    PubMed

    Chaivisuthangkura, Parin; Longyant, Siwaporn; Sithigorngul, Paisarn

    2014-02-12

    Among shrimp viral pathogens, white spot syndrome virus (WSSV) and yellow head virus (YHV) are the most lethal agents, causing serious problems for both the whiteleg shrimp, Penaeus (Litopenaeus) vannamei, and the black tiger shrimp, Penaeus (Penaeus) monodon. Another important virus that infects P. vannamei is infectious myonecrosis virus (IMNV), which induces the white discoloration of affected muscle. In the cases of taura syndrome virus and Penaeus stylirostris densovirus (PstDNV; formerly known as infectious hypodermal and hematopoietic necrosis virus), their impacts were greatly diminished after the introduction of tolerant stocks of P. vannamei. Less important viruses are Penaeus monodon densovirus (PmDNV; formerly called hepatopancreatic parvovirus), and Penaeus monodon nucleopolyhedrovirus (PemoNPV; previously called monodon baculovirus). For freshwater prawn, Macrobrachium rosenbergii nodavirus and extra small virus are considered important viral pathogens. Monoclonal antibodies (MAbs) specific to the shrimp viruses described above have been generated and used as an alternative tool in various immunoassays such as enzyme-linked immunosorbent assay, dot blotting, Western blotting and immunohistochemistry. Some of these MAbs were further developed into immunochromatographic strip tests for the detection of WSSV, YHV, IMNV and PemoNPV and into a dual strip test for the simultaneous detection of WSSV/YHV. The strip test has the advantages of speed, as the result can be obtained within 15 min, and simplicity, as laboratory equipment and specialized skills are not required. Therefore, strip tests can be used by shrimp farmers for the pond-side monitoring of viral infection. PMID:24567913

  6. Detection of shrimp infectious myonecrosis virus by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick.

    PubMed

    Puthawibool, Teeranart; Senapin, Saengchan; Kiatpathomchai, Wansika; Flegel, Timothy W

    2009-03-01

    Infectious myonecrosis virus (IMNV) has caused a slowly progressive disease with cumulative mortalities of up to 70% or more in cultured Penaeus (Litopenaeus) vannamei in Northeast Brazil and Indonesia. Rapid detection of viruses by loop-mediated isothermal amplification (LAMP) of genomic material with high specificity and sensitivity can be applied for diagnosis, monitoring and control of diseases in shrimp aquaculture. Using an IMNV template, successful detection was achieved after a 60-min RT-LAMP reaction using biotin-labeled primers followed by 5min hybridization with an FITC-labeled DNA probe and 5min assay using a chromatographic lateral flow dipstick (LFD). Thus, the combined system of RT-LAMP and LFD required a total assay interval of less than 75min, excluding the RNA extraction time. The sensitivity of detection was comparable to that of other commonly used methods for nested RT-PCR detection of IMNV. In addition to reducing amplicon detection time when compared to electrophoresis, LFD confirmed amplicon identity by hybridization and eliminated the need to handle carcinogenic ethidium bromide. The RT-LAMP-LFD method gave negative test results with nucleic acid extracts from normal shrimp and from shrimp infected with other viruses including infectious hypodermal hematopoietic necrosis virus (IHHNV), monodon baculovirus (MBV), a hepatopancreatic parvovirus from P. monodon (PmDNV), white spot syndrome virus (WSSV), yellow head virus (YHV), Taura syndrome virus (TSV), Macrobrachium rosenbergii nodavirus (MrNV) and gill associated virus (GAV). PMID:19022295

  7. Immunological-based assays for specific detection of shrimp viruses

    PubMed Central

    Chaivisuthangkura, Parin; Longyant, Siwaporn; Sithigorngul, Paisarn

    2014-01-01

    Among shrimp viral pathogens, white spot syndrome virus (WSSV) and yellow head virus (YHV) are the most lethal agents, causing serious problems for both the whiteleg shrimp, Penaeus (Litopenaeus) vannamei, and the black tiger shrimp, Penaeus (Penaeus) monodon. Another important virus that infects P. vannamei is infectious myonecrosis virus (IMNV), which induces the white discoloration of affected muscle. In the cases of taura syndrome virus and Penaeus stylirostris densovirus (PstDNV; formerly known as infectious hypodermal and hematopoietic necrosis virus), their impacts were greatly diminished after the introduction of tolerant stocks of P. vannamei. Less important viruses are Penaeus monodon densovirus (PmDNV; formerly called hepatopancreatic parvovirus), and Penaeus monodon nucleopolyhedrovirus (PemoNPV; previously called monodon baculovirus). For freshwater prawn, Macrobrachium rosenbergii nodavirus and extra small virus are considered important viral pathogens. Monoclonal antibodies (MAbs) specific to the shrimp viruses described above have been generated and used as an alternative tool in various immunoassays such as enzyme-linked immunosorbent assay, dot blotting, Western blotting and immunohistochemistry. Some of these MAbs were further developed into immunochromatographic strip tests for the detection of WSSV, YHV, IMNV and PemoNPV and into a dual strip test for the simultaneous detection of WSSV/YHV. The strip test has the advantages of speed, as the result can be obtained within 15 min, and simplicity, as laboratory equipment and specialized skills are not required. Therefore, strip tests can be used by shrimp farmers for the pond-side monitoring of viral infection. PMID:24567913

  8. Growth and antioxidant status of oriental river prawn Macrobrachium nipponense fed with diets containing vitamin E

    NASA Astrophysics Data System (ADS)

    Zhao, Weihong; Wang, Zisheng; Yu, Yebing; Qi, Zhitao; Lü, Linlan; Zhang, Yuxia; Lü, Fu

    2016-05-01

    A feeding trial was carried out to investigate the dietary vitamin E requirement of the oriental river prawn Macrobrachium nipponense (weight of 0.3-0.4 g) and its effect role on antioxidant activity. Prawns were fed with seven levels of vitamin E (0, 25, 50, 75, 100, 200, and 400 mg/kg diet) for 60 days. The results show that dietary vitamin E supplementation could significantly increased the prawn weight ( P < 0.05). The activity of superoxide dismutase (SOD) in the hepatopancreas was significantly higher in prawns fed with diets supplemented with ≤75 mg/kg vitamin E than in those fed with diets supplemented with 100-400 mg/kg vitamin E ( P < 0.05). The activity of catalase (CAT) in the hepatopancreas decreased significantly as dietary vitamin E supplementation increased ( P < 0.05), and no significant difference was detected in glutathione peroxidase (GSH-Px) activity between different dietary groups ( P >0.05). The contents of vitamin E in the hepatopancreas and in the muscle increased with increasing dietary vitamin E. There was a linear correlation between the vitamin E level in diet and that in muscle, and between the vitamin E level in diet and that in the hepatopancreas. All the above results indicated that dietary vitamin E can be stored in the hepatopancreas and muscle and lower both the activities of SOD and CAT in the hepatopancreas, suggesting that it is a potential antioxidant in M. nipponense. Broken line analysis conducted on the weight gains of prawns in each diet group showed that the dietary vitamin E requirement for maximum growth is 94.10 mg/kg.

  9. 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

  10. 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

  11. 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 α-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

  12. Growth and antioxidant status of oriental river prawn Macrobrachium nipponense fed with diets containing vitamin E

    NASA Astrophysics Data System (ADS)

    Zhao, Weihong; Wang, Zisheng; Yu, Yebing; Qi, Zhitao; Lü, Linlan; Zhang, Yuxia; Lü, Fu

    2015-11-01

    A feeding trial was carried out to investigate the dietary vitamin E requirement of the oriental river prawn Macrobrachium nipponense (weight of 0.3-0.4 g) and its effect role on antioxidant activity. Prawns were fed with seven levels of vitamin E (0, 25, 50, 75, 100, 200, and 400 mg/kg diet) for 60 days. The results show that dietary vitamin E supplementation could significantly increased the prawn weight (P<0.05). The activity of superoxide dismutase (SOD) in the hepatopancreas was significantly higher in prawns fed with diets supplemented with ≤75 mg/kg vitamin E than in those fed with diets supplemented with 100-400 mg/kg vitamin E (P<0.05). The activity of catalase (CAT) in the hepatopancreas decreased significantly as dietary vitamin E supplementation increased (P<0.05), and no significant difference was detected in glutathione peroxidase (GSH-Px) activity between different dietary groups (P>0.05). The contents of vitamin E in the hepatopancreas and in the muscle increased with increasing dietary vitamin E. There was a linear correlation between the vitamin E level in diet and that in muscle, and between the vitamin E level in diet and that in the hepatopancreas. All the above results indicated that dietary vitamin E can be stored in the hepatopancreas and muscle and lower both the activities of SOD and CAT in the hepatopancreas, suggesting that it is a potential antioxidant in M. nipponense. Broken line analysis conducted on the weight gains of prawns in each diet group showed that the dietary vitamin E requirement for maximum growth is 94.10 mg/kg.

  13. Population Structure and Historical Demography of the Oriental River Prawn (Macrobrachium nipponense) in Taiwan

    PubMed Central

    Chu, Ta-Jen; Wang, Daryi; Lee, Ying-Chou; Tzeng, Tzong-Der

    2015-01-01

    The oriental river prawn (Macrobrachium nipponense) is a non-obligatory amphidromous prawn, and it has a wide distribution covering almost the entire Taiwan. Mitochondrial DNA fragment sequences of the cytochrome oxidase subunit I (COI) and 16S rRNA were combined and used to elucidate the population structure and historical demography of oriental river prawn in Taiwan. A total of 202 individuals from six reservoirs and three estuaries were separately collected. Nucleotide diversity (π) of all populations was 0.01217, with values ranging from 0.00188 (Shihmen Reservoir, SMR, northern Taiwan) to 0.01425 (Mingte Reservoir, MTR, west-central Taiwan). All 76 haplotypes were divided into 2 lineages: lineage A included individuals from all sampling areas except SMR, and lineage B included specimens from all sampling locations except Chengching Lake Reservoir (CLR) and Liyu Lake Reservoir (LLR). All FST values among nine populations were significantly different except the one between Jhonggang River Estuary (JGE, west-central Taiwan) and Kaoping River Estuary (KPE, southern Taiwan). UPGMA tree of nine populations showed two main groups: the first group included the SMR and Tamsui River Estuary (TSE) (both located northern Taiwan), and the second one included the other seven populations (west-central, southern and eastern Taiwan). Demographic analyses implied a population expansion occurred during the recent history of the species. The dispersal route of this species might be from China to west-central and west-southern Taiwan, and then the part individuals belonging to lineage A and B dispersed southerly and northerly, respectively. And then part individuals in west-central Taiwan fell back to and stay at estuaries as the sea level rose about 18,000 years ago. PMID:26716687

  14. Population Structure and Historical Demography of the Oriental River Prawn (Macrobrachium nipponense) in Taiwan.

    PubMed

    Chen, Po-Cheng; Shih, Chun-Han; Chu, Ta-Jen; Wang, Daryi; Lee, Ying-Chou; Tzeng, Tzong-Der

    2015-01-01

    The oriental river prawn (Macrobrachium nipponense) is a non-obligatory amphidromous prawn, and it has a wide distribution covering almost the entire Taiwan. Mitochondrial DNA fragment sequences of the cytochrome oxidase subunit I (COI) and 16S rRNA were combined and used to elucidate the population structure and historical demography of oriental river prawn in Taiwan. A total of 202 individuals from six reservoirs and three estuaries were separately collected. Nucleotide diversity (π) of all populations was 0.01217, with values ranging from 0.00188 (Shihmen Reservoir, SMR, northern Taiwan) to 0.01425 (Mingte Reservoir, MTR, west-central Taiwan). All 76 haplotypes were divided into 2 lineages: lineage A included individuals from all sampling areas except SMR, and lineage B included specimens from all sampling locations except Chengching Lake Reservoir (CLR) and Liyu Lake Reservoir (LLR). All FST values among nine populations were significantly different except the one between Jhonggang River Estuary (JGE, west-central Taiwan) and Kaoping River Estuary (KPE, southern Taiwan). UPGMA tree of nine populations showed two main groups: the first group included the SMR and Tamsui River Estuary (TSE) (both located northern Taiwan), and the second one included the other seven populations (west-central, southern and eastern Taiwan). Demographic analyses implied a population expansion occurred during the recent history of the species. The dispersal route of this species might be from China to west-central and west-southern Taiwan, and then the part individuals belonging to lineage A and B dispersed southerly and northerly, respectively. And then part individuals in west-central Taiwan fell back to and stay at estuaries as the sea level rose about 18,000 years ago. PMID:26716687

  15. 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

  16. Molecular and functional characterization of the vitellogenin receptor in oriental river prawn, Macrobrachium nipponense.

    PubMed

    Bai, Hongkun; Qiao, Hui; Li, Fajun; Fu, Hongtuo; Jiang, Sufei; Zhang, Wenyi; Yan, Yuedi; Xiong, Yiwei; Sun, Shengming; Jin, Shubo; Gong, Yongsheng; Wu, Yan

    2016-04-01

    A complementary DNA (cDNA) that encodes the vitellogenin receptor (VgR) in the oriental river prawn, Macrobrachium nipponense, was cloned using expressed sequence tag analysis and a rapid amplification of cDNA ends approach. The coding region consists of 5920 base pairs (bp) that encode a 1902 amino acid protein, with a predicted molecular mass of 209kDa. The coding region is flanked by a 45bp 5'-untranslated region (UTR) and a 166bp 3'-UTR. The deduced amino acid sequence of the M. nipponense VgR cDNA had typically conserved domains, such as an extracellular, lipoprotein-binding domain, epidermal growth factor-like and O-glycosylation domains, a transmembrane domain and a short C-terminal, cytosolic tail. Quantitative real-time PCR (qPCR) indicated that Mn-VgR is highly expressed in the female ovary. Expression analysis by qPCR demonstrated the larval and ovarian developmental stage-specific expression pattern. As the ovaries developed, the expression level of Mn-VgR gradually increased during the reproductive cycle (stage I), to reach a peak in stage III. Levels then dropped as a new development cycle was entered after reproduction molting. Eyestalk ablation led to a significant increase in the expression of Mn-VgR during the ovarian development stages (P<0.05), when compared with the eyestalk-intact group. The investigation revealed that eyestalk ablation initially affected Mn-VgR expression and then influenced vitellogenesis. In adult females, VgR RNA interference (RNAi) dramatically delayed the maturation of the ovary, in accordance with the gonad somatic index. In addition, Mn-VgR RNAi led to vitellin depletion in the oocytes and the accumulation of vitellin in the hepatopancreas. PMID:26773480

  17. Osmoregulation in larvae and juveniles of two recently separated Macrobrachium species: Expression patterns of ion transporter genes.

    PubMed

    Boudour-Boucheker, Nesrine; Boulo, Viviane; Charmantier-Daures, Mireille; Anger, Klaus; Charmantier, Guy; Lorin-Nebel, Catherine

    2016-05-01

    In this comparative study, osmoregulatory mechanisms were analyzed in two closely related species of palaemonid shrimp from Brazil, Macrobrachium pantanalense and Macrobrachium amazonicum. A previous investigation showed that all postembryonic stages of M. pantanalense from inland waters of the Pantanal are able to hyper-osmoregulate in fresh water, while this species was not able to hypo-osmoregulate at high salinities. In M. amazonicum originating from the Amazon estuary, in contrast, all stages are able to hypo-osmoregulate, but only first-stage larvae, late juveniles and adults are able to hyper-osmoregulate in fresh water. The underlying molecular mechanisms of these physiological differences have not been known. We therefore investigated the expression patterns of three ion transporters (NKA α-subunit, VHA B-subunit and NHE3) following differential salinity acclimation in different ontogenetic stages (stage-V larvae, juveniles) of both species. Larval NKAα expression was at both salinities significantly higher in M. pantanalense than in M. amazonicum, whereas no difference was noted in juveniles. VHA was also more expressed in larvae of M. pantanalense than in those of M. amazonicum. When NHE3 expression is compared between the larvae of the two species, further salinity-related differences were observed, with generally higher expression in the inland species. Overall, a high expression of ion pumps in M. pantanalense suggests an evolutionary key role of these transporters in freshwater invasion. PMID:26872994

  18. Effects of Artificial Shelter and Chitosan on Growth Performance of Freshwater Prawn (Macrobrachium lanchesteri de Man, 1911).

    PubMed

    Jongyotha, Suriya; Doolgindachbaporn, Sompong; Suraniranat, Prawit

    2015-04-01

    Two experiments were carried out at the Department of Fisheries, Khon Kaen University, Khon Kaen, Thailand during September 2013 to February 2014 aiming to pursue both the better growth performance and survival rate of the freshwater prawns (Macrobrachium lanchesteri) where a Completely Randomized Design (CRD) with three replications was used for each experiment. For experiment 1, different percentages of grass shelter were used, i.e. 0% (Control, T1), 25% (T2), 50% (T3) and 75% (T4). For experiment 2, different percentages of commercial chitosan were used, i.e. 0% (Control, T1), 25% (T2), 50% (T3) and 75% (T4). Both experiments were carried out at the same time and each of them was lasted for 75 days. The results showed that the added grass shelter gave significantly higher Total Weight (TW) and Survival Rate (SR) than the control treatment. However, there were no significant differences found on TW and SR among the added grass shelter treatments (T2 up to T4). Chitosan gave significantly higher TW, AW and Number of Molting (NM) than the control treatment. There were no significant differences on TW, NM and AW found amongst the chitosan treatments. The results of the experiment 1 showed that the best rate of grass shelter for the culturing of the Macrobrachium lanchesteri was at a rate of 25% (T 2) and the best rate of the chitosan was at a rate of 4% (Diet 3) for the Experiment 2. PMID:26506647

  19. Reproductive and morphometric traits of Macrobrachium amazonicum (Decapoda: Palaemonidae) from the Pantanal, Brazil, suggests initial speciation.

    PubMed

    Hayd, Liliam; Anger, Klaus

    2013-03-01

    The palaemonid shrimp Macrobrachium amazonicum shows an unusually large geographic range (ca. 4000km across) living in coastal, estuarine, and limnic inland habitats of the upper Amazon, Orinoco, and La Plata basins. This raises doubts whether allopatric, ecologically diverse populations belong to the same species. While shrimps from estuarine and Amazonian habitats have been studied in great detail, very little is known about hololimnetic inland populations. In the present study, biological traits related to growth (maximum body size, fresh weight, morphometric relationships) and reproduction (sex ratio; occurrence of male morphotypes; minimum sexable size; minimum size of ovigerous females; fecundity; egg size), were studied in M amazonicum collected from a pond culture and two natural freshwater habitats (Rio Miranda; Lagoa Baiazinha) in the Pantanal of Mato Grosso do Sul, Brazil. In total, 2270 shrimps were examined (603 males; 1667 females, 157 of these ovigerous). Sex ratio (males:females) was at all sampling sites strongly female-biased, ranging from 0.2-0.6. Maximum body size was larger in natural habitats compared to the pond culture, suggesting reduced growth or a shorter life span under artificial mass rearing conditions. Maximum fecundity observed in our material was 676 eggs, reached by the largest female (TL=65mm; Lagoa Baiazinha). A significant difference between slope parameters of linear regressions describing fecundity, either in terms of numbers of eggs laid or of larvae released, in relation to female fresh weight, indicates egg losses. This may be due in part to a 2.4-fold increase in egg volume occurring during the course of embryonic development, while the available space under the abdomen remains limited. Size-weight relationships differed significantly between males and females, indicating sexspecific morphometric differences. Males appear to have a more slender body shape than females, reaching thus lower weight at equal TL. When reproductive and morphometric traits are compared with literature data from estuarine and inland populations living in the Amazon and Orinoco plains, shrimps from the Pantanal show conspicuous peculiarities differing from other populations: (1) maximum body size is far smaller, suggesting shorter longevity; (2) females are consistently larger than males; (3) different male morphotypes are absent; (4) minimum sexable size and (5) minimum size of ovigerous females are smaller. These traits suggest a heterochronic shift (predisplacement) of sexual maturation and r-selection. In summary, our data show biologically relevant differences in life-history traits of shrimps from the Pantanal compared to M. amazonicum populations in other regions. All these differences persist also in long-term cultures maintained under constant conditions. Altogether, our data support the hypothesis that M amazonicum in the Paraná-Paraguay drainage basin has phylogenetically diverged from allopatric populations that are hydrologically separated by continental watersheds, implying an at least incipient vicariant speciation. PMID:23894962

  20. Population structure of Macrobrachium australiense (Decapoda: Palaemonidae) in Western Queensland, Australia: the role of contemporary and historical processes.

    PubMed

    Carini, G; Hughes, J M

    2004-10-01

    Rivers of Western Queensland, Australia, represent a discontinuous and variable aquatic habitat for the freshwater fauna of the region. Rivers periodically fluctuate between being highly fragmented, with numerous disconnected waterholes and ephemeral channels, and being highly connected by a dominant system of anastomosing channels. We used mitochondrial sequences to investigate the genetic structure and inferred patterns of dispersal associated with this flow regime for the freshwater prawn, Macrobrachium australiense (Decapoda: Palaemonidae), sampling 28 localities throughout eight catchments. Based on a 505 base pair fragment of mitochondrial cytochrome c oxidase subunit I, we identified 98 haplotypes in a sample of 402 individuals. The haplotypes clustered into two main clades corresponding geographically to the major drainages: the Lake Eyre and Murray-Darling basins. Populations of M. australiense inhabiting the two basins appear to have diverged around 800,000 years ago (estimated sequence divergence of 1.6%). Analysis of population differentiation indicated contemporary high levels of genetic subdivision and restricted gene flow among populations within and among catchments. Phylogenetic analysis detected a series of historical range expansions in the region and we suggest that climate fluctuations during the Pleistocene have resulted in extensive floods that have promoted historical movements of aquatic organisms across catchment boundaries. PMID:15329659

  1. 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

  2. Molecular cloning and expression analysis of female sterile homeotic gene (fsh) in the oriental river prawn Macrobrachium nipponense.

    PubMed

    Zhang, W Y; Jiang, S F; Xiong, Y W; Fu, H T; Qiao, H; Sun, S M; Gong, Y S; Jin, S B

    2015-01-01

    The gene female sterile homeotic (fsh) plays crucial roles in molecular function, including protein kinase activity and DNA binding, which are involved in biological processes such as terminal region determination and negative regulation of DNA-dependent transcription. Although fsh has been found in Drosophila melanogaster, little is known regarding its expression in crustaceans. In this study, a fsh gene homologue, designated as Mnfsh, was cloned and characterized from the testis of the oriental river prawn, Macrobrachium nipponense, by using EST analysis and the RACE approach for the first time. The full-length cDNA of Mnfsh was 2029 bp, consisting of a 5' UTR of 361 bp, a 3' UTR of 216 bp, and an ORF of 1452 bp encoding 484 amino acids. qRT-PCR analysis showed that the Mnfsh gene was expressed in the testis, ovary, muscle, heart, eyestalk, and abdominal ganglion, with the highest level of expression in the ovary and the lowest in the heart. qRT-PCR analyses showed that the expression levels of Mnfsh mRNA both significantly increased in the zoea stage, the VII larvae, and 1st day post-larvae after metamorphosis. In conclusion, the results of the present study indicate that Mnfsh is an arthropod fsh homologue and probably also plays important roles in embryogenesis, organogenesis, and morphological differentiation of M. nipponense. PMID:25966204

  3. 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

  4. Light-mediated DNA Repair Prevents UVB-induced Cell Cycle Arrest in Embryos of the Crustacean Macrobrachium olfersi.

    PubMed

    Zeni, Eliane Cristina; Ammar, Dib; Leal, Mayana Lacerda; da Silva, Heloisa Schramm; Allodi, Silvana; Müller, Yara Maria Rauh; Nazari, Evelise Maria

    2015-01-01

    High levels of ultraviolet-B (UVB) radiation can negatively affect aquatic animals. Macrobrachium olfersi is a prawn that lives in clear freshwaters and during the breeding season, females carry eggs in an external brood pouch. Therefore, we hypothesize that eggs are also exposed to environmental UVB radiation. The aim of this study was to investigate whether UVB radiation induces DNA damage and compromises cell cycle in embryos of M. olfersi. In laboratory, UVB irradiance (310 mW. cm(-2) ) that embryos receive in the natural environment was simulated. After irradiation, embryos were kept under different light conditions in order to recognize the presence of cell repair. UVB radiation induces DNA damage, specifically thymine dimers. After 48 h of UVB exposure, a significant decrease in the level of these dimers was observed in embryos kept under visible light while it remained constant in the dark. Moreover, under visible light and darkness, a decrease in proliferation was observed after 48 h of irradiation. An increase in PCNA expression and decrease in p53 expression were observed after, respectively, 1 and 48 h of exposure. Our results showed that UVB radiation disturbs the cell cycle and induces DNA damage in M. olfersi embryos. However, under visible light these embryos showed successful DNA repair. PMID:25869065

  5. Regulation of essential heavy metals (Cu, Cr, and Zn) by the freshwater prawn macrobrachium malcolmsonii (Milne Edwards)

    SciTech Connect

    Vijayram, K.; Geraldine, P.

    1996-02-01

    Despite the low concentrations of heavy metals in the surrounding medium, aquatic organisms take them up and accumulate them in their soft tissues to concentrations several fold higher than those of ambient levels. Knowledge of accumulation patterns of a particular trace metal is a prerequisite for understanding the significance of an observed metal concentration in a particular animal, especially from the aspect of biomonitoring. Many marine invertebrates accumulate heavy metals without any regulation and the accumulation necessarily being associated with mechanisms to store the metals in a detoxified form. Two detoxification mechanisms have been described, both of which may occur in one specimen. Heavy metals can either be bound up in insoluble metalliferous {open_quote}granules{close_quote}, or are bound to soluble metal-binding ligands, such as metallothioneins. Some marine decapod crustaceans have an innate ability to regulate the internal concentrations of essential but potentially toxic metals within a constant level, presumably to meet their metabolic demands. However, at present, there is no such information relating to freshwater decapod crustaceans, especially shrimps which occupy a totally different environment. Macrobrachium malcolmsonii, a potential aquaculture species for freshwater is found in abundance in one of the major Indian rivers, the Cauvery. In the present study, an attempt was made to determine whether the freshwater prawn, M. malcolmsonlii, is able to regulate the three essential elements, copper, chromium and zinc, over a wide range of dissolved concentrations. These three metals were chosen because the Cauvery River receives pollutants containing these metals.

  6. Gene expression profile analysis of testis and ovary of oriental river prawn, Macrobrachium nipponense, reveals candidate reproduction-related genes.

    PubMed

    Qiao, H; Xiong, Y W; Jiang, S F; Fu, H T; Sun, S M; Jin, S B; Gong, Y S; Zhang, W Y

    2015-01-01

    This study utilized high-throughput RNA sequencing technology to identify reproduction- and development-related genes of Macrobrachium nipponense by analyzing gene expression profiles of testis and ovary. More than 20 million 1 x 51-bp reads were obtained by Illumina sequencing, generating more than 7.7 and 11.7 million clean reads in the testis and ovary library, respectively. As a result, 10,018 unitags were supposed to be differentially expressed genes (DEGs) between ovary and testis. Compared to the ovary library, 4563 (45.5%) of these DEGs exhibited at least 6-fold upregulated expression, while 5455 (54.5%) DEGs exhibited at least 2-fold downregulated expression in the testis. The Gene Ontology (GO) enrichment analysis showed that 113 GO terms had potential molecular functions in reproduction. The Kyoto Encyclopedia of Genes and Genomes results revealed that the most important pathways may be relevant to reproduction and included 7 pathways. Forty-two genes were identified as reproduction-, development-, and sex-related genes based on GO classification and sequence comparison with other publications, including male reproductive-related LIM protein, spermatogenesis-associated protein, gametocyte-specific factor 1, VASA-like protein, vitellogenin, sex-determining protein fem-1, and other potential candidates. These results will advance research in the field of molecular genetics in M. nipponense and offer a valuable resource for further research related to reproduction in crustaceans. PMID:25867350

  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 analysis of the freshwater prawn Macrobrachium olfersii (Decapoda, Palaemonidae) supports the existence of a single species throughout its distribution.

    PubMed

    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

  9. 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

  10. Constructing and random sequencing analysis of normalized cDNA library of testis tissue from oriental river prawn (Macrobrachium nipponense).

    PubMed

    Qiao, Hui; Fu, Hongtuo; Jin, Shubo; Wu, Yan; Jiang, Sufei; Gong, Yongsheng; Xiong, Yiwei

    2012-09-01

    The oriental river prawn, Macrobrachium nipponense, is an important aquaculture species in China. Sexual precocity is a serious problem because of genetic retrogression, which has negative effects on product quality and dramatically affects price. Culture of all-male populations of this species would be economically advantageous, as the males grow faster and reach a much larger size than females. Developing such a culture scheme will require discovery of sex- or reproduction-related genes that affect sexual maturity and sex determination. In this study, a high-quality normalized testis cDNA library was constructed to identify novel transcripts. Of the 5280 successful sequencing reaction yields, 5202 expressed tagged sequences (ESTs) with an average length of 954 bp. Ultimately, 3677 unique sequences, including 891 contigs and 2786 singletons, were identified based on cluster and assembly analyses. Sixteen hundred (43.5%) genes were novel based on the NCBI protein database, thus these unidentified genes may improve basic molecular knowledge about M. nipponense. Of the novel unigenes, 34.4% (715/2077) were homologous to insects, such as Tribolium castaneum, Drosophila spp. and Apis mellifera. Fifty-two genes were identified as sex- or reproduction-related based on Gene Ontology classification and sequence comparison with data from other publications. These genes can be classified into groups based on different functions, including 10 sex-determination related genes, 8 male-reproductive genes, 5 cathepsin-related genes, 20 ubiquitin-related genes, 5 ferritin-related genes, and 4 LRR genes. The results of this study provide new sequence information about M. nipponense, which will be the basis for further genetic studies of this species and other decapods crustaceans. PMID:22632994

  11. Molecular cloning, characterization, and expression analysis of two different types of lectins from the oriental river prawn, Macrobrachium nipponense.

    PubMed

    Xiu, Yunji; Wang, Yinghui; Jing, Yunting; Qi, Yakun; Ding, Zhengfeng; Meng, Qingguo; Wang, Wen

    2015-08-01

    Lectins, which are widely expressed in invertebrates, play important roles in many biological processes, including protein trafficking, cell signaling, pathogen recognition, as effector molecules, and so on (Wang and Wang, 2013). This study identified one novel M-type lectin and one L-Type lectin, designated as MnMTL1 and MnLTL1, from the oriental river prawn Macrobrachium nipponense. The full-length cDNA of MnMTL1 was 2064 bp with a 1761 bp ORF encoding a putative protein of 586 deduced amino acids. The full-length cDNA of MnLTL1 was 1744 bp with a 972 bp ORF encoding a 323-amino acid peptide. The deduced MnMTL1 protein contained a putative type II transmembrane region and a 440-aa Glycoside hydrolase family 47 (GH47) domain. One luminal carbohydrate recognition domain and a 23-aa type I transmembrane region were identified from the MnLTL1. MnMTL1 shared 78% identity with Marsupenaeus japonicus M-type lectin and MnLTL1 shared 83% similarity with M. japonicus L-type lectin. RT-PCR analysis showed that MnMTL1 and MnLTL1 were expressed in all tested tissues. Quantitative real-time PCR analysis revealed that MnMTL1 and MnLTL1 are substantially fluctuant during Aeromonas hydrophila and Aeromonas veronii infections. Based on immune responses and previous literature, we assumed that MnMTL1 and MnLTL1 might be functioned as pattern recognition receptors and play important roles in the immune response of M. nipponense. PMID:25929240

  12. Comparative transcriptome analysis reveals molecular strategies of oriental river prawn Macrobrachium nipponense in response to acute and chronic nitrite stress.

    PubMed

    Xu, Zhixin; Li, Tongyu; Li, Erchao; Chen, Ke; Ding, Zhili; Qin, Jian G; Chen, Liqiao; Ye, Jinyun

    2016-01-01

    Macrobrachium nipponense is an economically and nutritionally important species threatened by ambient superfluous nitrite. De novo RNA-Seq was used to explore the molecular mechanism in M. nipponense exposed to the acute nitrite stress (26.05 mg/L nitrite-N) for 24 h and the chronic nitrite stress (1.38 mg/L nitrite-N) for 28 d A total of 175.13 million reads were obtained and assembled into 58,871 unigenes with an average length of 1028.7 bp and N50 of 1294 bp. Under the acute and chronic nitrite stress trials, 2824 and 2610 unigenes were significantly expressed. In GO analysis and KEGG pathway analysis, 30 pathways were significantly different between the two treatments while four pathways were in common and the markedly altered pathways were divided into four sections as immunity, metabolism, cell and others. The immunity section revealing the different depth of immunity provoked by nitrite stress contained the most pathways including the important pathways as phagosome, folate biosynthesis, glycerolipid metabolism, glycine, serine and threonine metabolism, amino sugar and nucleotide sugar metabolism under the acute nitrite stress, and lysosome, alanine, aspartate and glutamate metabolism, arginine and proline metabolism under the chronic nitrite stress. This is the first report of responses of M. nipponense under acute and chronic nitrite stress through de novo transcriptome sequencing on the transcriptome level. The results of transcriptome analysis improve our understanding on the underlying molecular mechanisms coping with nitrite stress in crustacean species. PMID:26687531

  13. 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

  14. A kinetic characterization of the gill V(H(+))-ATPase in juvenile and adult Macrobrachium amazonicum, a diadromous palaemonid shrimp.

    PubMed

    Lucena, Malson N; Pinto, Marcelo R; Garçon, Daniela P; McNamara, John C; Leone, Francisco A

    2015-03-01

    Novel kinetic properties of a microsomal gill V(H(+))-ATPase from juvenile and adult Amazon River shrimp, Macrobrachium amazonicum, are described. While protein expression patterns are markedly different, Western blot analysis reveals a sole immunoreactive band, suggesting a single V(H(+))-ATPase subunit isoform, distributed in membrane fractions of similar density in both ontogenetic stages. Immunofluorescence labeling locates the V(H(+))-ATPase in the apical regions of the lamellar pillar cells in both stages in which mRNA expression of the V(H(+))-ATPase B-subunit is identical. Juvenile (36.6±3.3 nmol Pi min(-1) mg(-1)) and adult (41.6±1.3 nmol Pi min(-1) mg(-1)) V(H(+))-ATPase activities are similar, the apparent affinity for ATP of the adult enzyme (K0.5=0.21±0.02 mmol L(-1)) being 3-fold greater than for juveniles (K0.5=0.61±0.01 mmol L(-1)). The K0.5 for Mg(2+) interaction with the juvenile V(H(+))-ATPase (1.40 ± 0.07 mmol L(-1)) is ≈6-fold greater than for adults (0.26±0.02 mmol L(-1)) while the bafilomycin A1 inhibition constant (KI) is 45.0±2.3 nmol L(-1) and 24.2±1.2 nmol L(-1), for juveniles and adults, respectively. Both stages exhibited residual bafilomycin-insensitive ATPase activity of ≈25 nmol Pi min(-1) mg(-1), suggesting the presence of ATPases other than the V(H(+))-ATPase. These differences may reflect a long-term regulatory mechanism of V(H(+))-ATPase activity, and suggest stage-specific enzyme modulation. This is the first kinetic analysis of V(H(+))-ATPase activity in different ontogenetic stages of a freshwater shrimp and allows better comprehension of the biochemical adaptations underpinning the establishment of palaemonid shrimps in fresh water. PMID:25461614

  15. 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.

  16. Genomic cloning, expression, and single nucleotide polymorphism association analysis of the insulin-like androgenic gland hormone gene in the oriental river prawn (Macrobrachium nipponense).

    PubMed

    Li, F J; Jiang, F W; Bai, H K; Fu, H T; Jin, S B; Sun, S M; Qiao, H; Zhang, W Y

    2015-01-01

    Increasing evidence suggests that the insulin-like androgenic gland hormone (IAG) gene plays an important role in male sexual differentiation, metabolism, and growth in crustaceans. In the present study, we isolated the full-length genome sequence of IAG by genome walking based on the cDNA sequence in Macrobrachium nipponense. Four novel single nucleotide polymorphisms (SNPs) were studied, including 509G>T, 529G>T, 590A>T in intron 1, and 2226A>G in intron 2. The association of genetic variation with growth traits [body length (BL) and body weight (BW)] was analyzed. Individuals with GG geno- type at locus 2226A>G maintained higher mean BL (P < 0.01) and BW (P < 0.05) than AA and GA individuals. These results suggest that IAG SNPs may be useful molecular markers for selecting growth traits in M. nipponense. PMID:26125790

  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. 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

  19. 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

  20. Identification of androgenic gland microRNA and their target genes to discover sex-related microRNA in the oriental river prawn, Macrobrachium nipponense.

    PubMed

    Jin, S B; Fu, H T; Jiang, S F; Xiong, Y W; Qiao, H; Zhang, W Y; Gong, Y S; Wu, Y

    2015-01-01

    The oriental river prawn, Macrobrachium nipponense, is an important aquaculture species in China. The androgenic gland produces hormones that play crucial roles in the differentiation of crustaceans to the male sex. MicroRNA (miRNA) post-transcriptionally regulates many protein-coding genes, influencing important biological and metabolic processes. However, currently, there is no published data identifying miRNA in M. nipponense. In this study, we identified novel miRNA in the androgenic gland of M. nipponense. Using the high-throughput Illumina Solexa system, 1077 miRNA were identified from small RNA libraries by aligning with the de novo androgenic gland transcriptome of M. nipponense (obtained from RNA-Seq) and the sequences in the miRBase21 database. A total of 8,248, 76,011, and 78,307 target genes were predicted in the EST and SRA sequences provided in the NCBI database, and the androgenic gland transcriptome of M. nipponense, respectively. Some potential sex-related miRNA were identified based on the function of the predicted target genes. The results of our study provide new information regarding the miRNA expression in M. nipponense, which could be the basis for further genetic studies on decapod crustaceans. PMID:26782487

  1. Molecular cloning, characterization, and expression analysis of a Broad-Complex homolog during development in the oriental river prawn Macrobrachium nipponense.

    PubMed

    Jiang, S F; Zhang, Y P; Sun, S M; Gong, Y S; Xiong, Y W; Qiao, H; Zhang, W Y; Jin, S B; Fu, H T

    2015-01-01

    Broad-Complex (BR-C) is an early ecdysone-responsive gene encoding a family of zinc-finger transcription factors. In this study, we isolated the full-length cDNA of a BR-C homolog from the testes of the oriental river prawn (Macrobrachium nipponense), according to established expressed sequence tag information, using the rapid amplification of cDNA ends technique. The homolog was designated as MnBR-C. The full-length cDNA of MnBR-C contained a 1095-bp open reading frame encoding a precursor protein of 365 amino acid residues. Comparative and bioinformatic analyses revealed that MnBR-C exhibited a high degree of homology with BR-C proteins, and contained the BTB and Zf-H2C2-2 domains. Real-time quantitative polymerase chain reaction (qPCR) analysis revealed that the MnBR-C expression level varied significantly in the developing embryo, postembryonic larva, and adult tissue. Real-time qPCR showed that the MnBR-C gene was expressed in all of the tissues investigated, with the highest level of expression in the brain. In addition, MnBR-C was more abundantly expressed in the testes than in the ovaries. PMID:26125707

  2. Use of the Multispecies Freshwater Biomonitor to assess behavioral changes of Poecilia reticulata (Cyprinodontiformes: Poeciliidae) and Macrobrachium lanchesteri (Decapoda: Palaemonidae) in response to acid mine drainage: laboratory exposure.

    PubMed

    Mohti, Azmah; Shuhaimi-Othman, Mohammad; Gerhardt, Almut

    2012-09-01

    The behavioral responses of guppy Poecilia reticulata (Poeciliidae) and prawn Macrobrachium lanchesteri (Palaemonidae) individuals exposed to acid mine drainage (AMD) were monitored online in the laboratory with a Multispecies Freshwater Biomonitor™ (MFB). These responses were compared to those to reference water acidified to the respective pH values (ACID). Test animals in the juvenile stage were used for both species and were exposed to AMD and ACID for 24 hours. The stress behaviors of both test animals consisted mainly of decreased activity in AMD and increased activity in ACID, indicating that the metals in the AMD played a role as a stress factor in addition to pH. The locomotor activity levels of guppies and prawns for the ACID treatment were higher than the locomotor activity levels for the AMD treatment with increasing pH value. For guppies, significant differences were observed when specimens were exposed to AMD and ACID at pH 5.0 and 6.0; the percentage activities were only 16% and 12%, respectively, for AMD treatment, whereas for ACID treatment, the percentage activities were 35% and 40%, respectively, similar to the value of 36% for the controls. Similar trends were also observed for prawns, for which the percentage activities were only 6% and 4%, respectively, for AMD treatment, whereas for ACID treatment, the percentage activities were 31% and 38%, respectively, compared to 44% in the controls. This study showed that both species are suitable for use as indicators for ecotoxicity testing with the MFB. PMID:22868673

  3. 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

  4. 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…

  5. 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 μg/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

  6. An evaluation of replacing fish meal with fermented soybean meal in the diet of Macrobrachium nipponense: Growth, nonspecific immunity, and resistance to Aeromonas hydrophila.

    PubMed

    Ding, Zhili; Zhang, Yixiang; Ye, Jinyun; Du, Zhenyu; Kong, Youqin

    2015-05-01

    Partial or complete replacement of fish meal (FM) with fermented soybean meal (FSM) was examined in Macrobrachium nipponense over an 8-week growth trial. Growth and immune characteristics were evaluated. Fermented soybean meal replaced 0 (FM, control), 25% (R25), 50% (R50), 75% (R75), or 100% of the FM (R100) in five isocaloric and isonitrogenous diets. Each diet was fed to juvenile prawns (mean weight, 0.103 ± 0.0009 g) twice daily to apparent satiation in five replicates. Weight gain and specific growth rate of M. nipponense were significantly higher in prawns fed the R25 diet than that of prawns fed the FM diet. No significant differences were observed among the other treatments. Total hemocyte count and hemolymph phagocytic activity decreased as the proportion of FSM increased. Total antioxidant activity competence and malondialdehyde level in the hepatopancreas were highest in prawns fed the R100 diet. mRNA levels of the antioxidant genes Cu-Zn superoxide dismutase and catalase, heat shock cognate protein 70, and heat shock protein 90 were significantly differentially regulated in the prawn hepatopancreas. In addition, percent mortality increased after challenge with live Aeromonas hydrophila. Percent mortality of prawns fed the R100 diet was significantly higher than that of prawns fed the FM and R25 diets. These findings demonstrate that (1) M. nipponense growth performance was not affected by including a high proportion of FSM in the diet, and the best growth performance was obtained when 25% of the FM was replaced with FSM; (2) nonspecific immunity was impaired when all of the FM was replaced with FSM. PMID:25707598

  7. 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

    2015-10-01

    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 μg/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

  8. Identification and comparative analysis of the oriental river prawn (Macrobrachium nipponense) microRNA expression profile during hypoxia using a deep sequencing approach.

    PubMed

    Sun, Shengming; Fu, Hongtuo; Ge, Xianping; Zhu, Jian; Gu, Zhimin; Xuan, Fujun

    2016-03-01

    Hypoxia refers to a state of oxygen deficiency, which is observed frequently in aquaculture ponds. MicroRNAs (miRNAs) are small non-coding RNAs that are important effectors in regulating gene expression through posttranscriptional mechanisms. They are key elements in the response to hypoxia. The oriental river prawn (Macrobrachium nipponense) is an important commercial aquaculture species, and is sensitive to hypoxia. To date, there are no reports describing M. nipponense miRNAs. In this study, Solexa deep sequencing technology was used for high-throughput analysis of miRNAs in a small RNA library isolated from four M. nipponense tissues (gill, hepatopancreas, muscle and hemocytes). In total, 9,227,356 reads were obtained, 4,293,155 of which were related to 267 unique miRNAs, including 203 conserved and 64 prawn-specific miRNAs. Furthermore, miRNA features including length distribution and end variations were characterized. Annotation of targets revealed a broad range of biological processes and signal transduction pathways regulated by M. nipponense miRNAs. In addition, 880 co-expressed and 39 specific (25 normoxia-specific and 14 hypoxia-specific) miRNAs that may be involved in the response to hypoxia were confirmed using miRNA microarray analysis from the four prawn tissues combined. Real-time quantitative PCR (qPCR) analysis of eight miRNAs in the normoxia and hypoxia groups showed good concordance between the sequencing and qPCR data. This study provides the first large-scale identification and characterization of M. nipponense miRNAs and their potential targets, and represents a foundation for further characterization of their roles in the regulation of the diversity of hypoxia processes. PMID:26829004

  9. Molecular characterization and developmental expression of vitellogenin in the oriental river prawn Macrobrachium nipponense and the effects of RNA interference and eyestalk ablation on ovarian maturation.

    PubMed

    Bai, Hongkun; Qiao, Hui; Li, Fajun; Fu, Hongtuo; Sun, Shengming; Zhang, Wenyi; Jin, Shubo; Gong, Yongsheng; Jiang, Sufei; Xiong, Yiwei

    2015-05-10

    Vitellogenin (Vg) is the precursor of yolk protein, which functions as a nutritive resource that is important for embryonic growth and gonad development. In this study, the cDNA encoding the Vg gene from the oriental river prawn Macrobrachium nipponense was cloned using expressed sequence tag (EST) analysis and the rapid amplification of cDNA ends (RACE) approach. The transcript encoded 2536 amino acids with an estimated molecular mass of 286.810 kDa. Quantitative real-time PCR indicated high expression of Mn-Vg in the female ovary, hemocytes, and hepatopancreas. As ovaries developed, the expression level of Mn-Vg increased in both the hepatopancreas and ovary. In the hepatopancreas, the expression level rose more slowly at the early stage of vitellogenesis and reached the peak more rapidly compared to the expression pattern in ovary. The observed changes in Mn-Vg expression level at different development stages suggest the role of nutrient source in embryonic and larval development. Eyestalk ablation caused the Mn-Vg expression level to increase significantly compared to eyestalk-intact groups during the ovary development stages. Ablation accelerated ovary maturation by removing hormone inhibition of Mn-Vg in the hepatopancreas and ovary. In adult females, Mn-Vg dsRNA injection resulted in decreased expression of Mn-Vg in both the hepatopancreas and ovary, and two injection treatment dramatically delayed ovary maturation. Vg RNA interference down-regulated the vitellogenin receptor (VgR) expression level in the ovary, which illustrates the close relationship between Vg and VgR in the process of vitellogenesis. PMID:25499697

  10. Characterization, expression, and function analysis of gonad-inhibiting hormone in Oriental River prawn, Macrobrachium nipponense and its induced expression by temperature.

    PubMed

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

    2015-07-01

    Gonad-inhibiting hormone (GIH) is a member of crustacean hyperglycemic hormone family and plays a major role in regulating reproduction in crustaceans. In this study, a full-length cDNA of GIH of Oriental River prawn, Macrobrachium nipponense (Mn-GIH) was cloned from the eyestalk. A 1350 bp full-length Mn-GIH cDNA harbored 336 bp of an open reading frame encoding signal peptide of 112 amino acid residues. Sequence analysis revealed that the overall cDNA sequence and specific functional sites of Mn-GIH were highly conserved with those in other crustacean species. Expression analysis by quantitative real-time PCR demonstrated its tissue-specific, larval developmental stage-specific, and ovary developmental stage-specific expression pattern, respectively. The RNAi by GIH-ds-RNA in vivo injection was effective in this study and resulted a 50% (day 1), 83% (day 5) and 63% (day 9) down-regulation compared to control. The obvious changes of gonad somatic index (GSI) rate also provided strong evidence to the inhibition effects of GIH on ovary maturation and spawning. Four temperature gradients (12 °C ± 1 °C, 17 °C ± 1 °C, 22 °C ± 1 °C, 27 °C ± 1 °C) were set to imitate the temperature in breeding and non-breeding seasons. The observed expression profiles suggest that Mn-GIH did not display a high level expression as supposed to maintain an immature ovary state under low temperature (12 °C). The results indicated that GIH was probably activated to concentrating and working by a proper temperature before reaching to breeding season. PMID:25770669

  11. Immunocytochemical localization of V-H(+) -ATPase, Na(+) /K(+) -ATPase, and carbonic anhydrase in gill lamellae of adult freshwater euryhaline shrimp Macrobrachium acanthurus (Decapoda, Palaemonidae).

    PubMed

    Maraschi, Anieli Cristina; Freire, Carolina Arruda; Prodocimo, Viviane

    2015-08-01

    Physiological (organismal), biochemical, and molecular biological contributions to the knowledge of the osmoregulatory plasticity of palaemonid freshwater shrimps has provided a fairly complete model of transporter localization in their branchial epithelium. Direct immunological demonstration of the main enzymes in the gill epithelia of adult palaemonids is, however, still incipient. The diadromous freshwater shrimp Macrobrachium acanthurus was exposed to increased salinity (25‰ for 24 hr), and its responses at the systemic level were evaluated through the assays of hemolymph osmolality and muscle hydration, and at cellular and subcellular levels through the activity and localization of the V-H(+) -ATPase, the Na(+) /K(+) -ATPase, and the carbonic anhydrase. Results showed an increase in hemolymph osmolality (629 ± 5.3 mOsm/kg H2 O) and a decrease in muscle hydration (73.8 ± 0.5%), comparing values after 24 hr in 25‰ with control shrimps in freshwater (respectively 409.5 ± 15.8 mOsm/kg H2 O and 77.5 ± 0.4%). V-H(+) -ATPase was localized in pillar cells, whereas Na(+) /K(+) -ATPase in the septal cells. The main novelty of this study was that carbonic anhydrase was localized in the whole branchial tissue, in pillar and septal cells. Exposure to high salinity for 24 hr led to no detectable changes in their localization or in vitro activity. Immunolocalization data corroborated the literature and current models of palaemonid gill ion transport. The absence of changes reinforces the need for the constant expression of these enzymes to account for the euryhalinity of these shrimps. PMID:26036663

  12. Molecular characterization and mRNA expression of hypoxia inducible factor-1 and cognate inhibiting factor in Macrobrachium nipponense in response to hypoxia.

    PubMed

    Sun, Shengming; Xuan, Fujun; Fu, Hongtuo; Ge, Xianping; Zhu, Jian; Qiao, Hui; Jin, Shubo; Zhang, Wenyi

    2016-01-01

    Hypoxia inducible factors (HIFs) are considered to be the master switches of oxygen-dependent gene expression in mammalian species. Currently, very little is known about the function of this important pathway or the molecular structures of key players in the hypoxia-sensitive Oriental River Prawn Macrobrachium nipponense. In this study, HIFs-1α (HIF-1α), -1β (HIF-1β) and HIF 1-alpha inhibitor (FIH-1) from M. nipponense were cloned. The 4903-bp cDNA of M. nipponense HIF-1α (MnHIF-1α) encodes a protein of 1088 aa, M. nipponense HIF-1β (MnHIF-1β) spans 2042bp encoding 663 aa and the 1163bp M. nipponense FIH-1 (MnFIH-1) specifies a polypeptide of 345 aa. MnHIF-1 and MnFIH-1 homologs exhibit significant sequence similarity and share key functional domains with previously described vertebrate and invertebrate isoforms. Phylogenetic analysis identifies that genetic diversification of HIF-1 and FIH-1 occurred within the invertebrate lineage, indicating functional specialization of the oxygen sensing pathways in this group. Quantitative real-time RT-PCR demonstrated that MnHIF-1 and MnFIH-1 mRNA are expressed in different tissues and exhibit transcriptional responses to severe hypoxia in gill and muscle tissue, consistent with their putative role in oxygen sensing and the adaptive response to hypoxia. The role of HIF-1α in response to hypoxia was further investigated in the gills and muscles of prawns using in situ hybridization. These results suggested that HIF-1α plays an important role in oxygen sensing and homeostasis in M. nipponense. PMID:26883381

  13. Effect of salinity on the metabolism and osmoregulation of selected ontogenetic stages of an Amazon population of Macrobrachium amazonicum shrimp (Decapoda, Palaemonidae).

    PubMed

    Mazzarelli, C C M; Santos, M R; Amorim, R V; Augusto, A

    2015-05-01

    Probably as a function of their wide geographical distribution, the different population of Macrobrachium amazonicum shrimp may present distinct physiological, biochemical, reproductive, behavioral, and ecological patterns. These differences are so accentuated that the existence of allopatric speciation has been suggested, although initial studies indicate that the genetic variability of populations happen at an intraspecific level. Among the biological responses described for M. amazonicum populations, those regarding osmoregulation and metabolism play a key role for being related to the occupation of diverse habitats. To this effect, we investigated osmoregulation through the role of free amino acids in cell volume control and metabolism, through oxygen consumption in larvae (zoeae I, II, V and IX) and/or post-larvae of a M. amazonicum population from Amazon, kept in aquaculture fish hatcheries in the state of São Paulo. The results add information regarding the existence of distinct physiological responses among M. amazonicum populations and suggest that possible adjustments to metabolism and to the use of free amino acids as osmolytes of the regulation of the larvae and post-larvae cell volume depend on the appearance of structures responsible for hemolymph osmoregulation like, for example, the gills. In this respect, we verified that zoeae I do not alter their metabolism due to the exposition to fresh or brackish water, but they reduce intracellular concentration of free amino acids when exposed to fresh water, what may suggest the inexistence or inefficient performance of the structures responsible for volume regulation and hemolymph composition. On the other hand, in zoeae II and V exposed to fresh and brackish water, metabolism alterations were not followed by changes in free amino acids concentration. Thus it is possible, as the structures responsible for osmoregulation and ionic regulation become functional, that the role of free amino acids gets diminished and oxygen consumption elevated, probably due to greater energy expenditure with the active transportation of salts through epithelial membranes. Osmotic challenges also seem to alter throughout development, given that in zoeae II oxygen consumption is elevated on brackish water of 18, but in zoeae V it happens in fresh water. After M. amazonicum metamorphosis, free amino acids begin to play an important role as intracellular osmolytes, because we verified an increase of up to 40% in post-larvae exposed to brackish water of 18. The main free amino acids involved in cell volume regulation of ontogenetic stages evaluated were the non essential ones: glutamic acid, glycine, alanine, arginine, and proline. Interestingly, larvae from estuarine population studied here survived until the zoeae V stage in fresh water, but in some populations far from the sea, zoeae die right after eclosion in fresh water or they do not reach zoeae III stage. In addition, given that in favorable conditions caridean shrimp larvae shorten their development, we may infer that the cultivation environment, in which larvae developed in the present work, was appropriate, because almost all zoeae VIII kept on brackish water underwent metamorphosis directly to post-larvae and did not go through zoeae IX stage. PMID:26132021

  14. Differential adjustment in gill Na+/K+- and V-ATPase activities and transporter mRNA expression during osmoregulatory acclimation in the cinnamon shrimp Macrobrachium amazonicum (Decapoda, Palaemonidae).

    PubMed

    Faleiros, Rogério Oliveira; Goldman, Maria Helena S; Furriel, Rosa P M; McNamara, John Campbell

    2010-11-15

    We evaluate osmotic and chloride (Cl(-)) regulatory capability in the diadromous shrimp Macrobrachium amazonicum, and the accompanying alterations in hemolymph osmolality and [Cl(-)], gill Na(+)/K(+)-ATPase activity, and expression of gill Na(+)/K(+)-ATPase α-subunit and V-ATPase B subunit mRNA during salinity (S) acclimation. We also characterize V-ATPase kinetics and the organization of transport-related membrane systems in the gill epithelium. Macrobrachium amazonicum strongly hyper-regulates hemolymph osmolality and [Cl(-)] in freshwater and in salinities up to 25‰ S. During a 10-day acclimation period to 25‰ S, hemolymph became isosmotic and hypo-chloremic after 5 days, [Cl(-)] alone remaining hyporegulated thereafter. Gill Na(+)/K(+)-ATPase α-subunit mRNA expression increased 6.5 times initial values after 1 h, then decreased to 3 to 4 times initial values by 24 h and to 1.5 times initial values after 10 days at 25‰ S. This increased expression was accompanied by a sharp decrease at 5 h then recovery of initial Na(+)/K(+)-ATPase activity within 24 h, declining again after 5 days, which suggests transient Cl(-) secretion. V-ATPase B-subunit mRNA expression increased 1.5-fold within 1 h, then reduced sharply to 0.3 times initial values by 5 h, and remained unchanged for the remainder of the 10-day period. V-ATPase activity dropped sharply and was negligible after a 10-day acclimation period to 21‰ S, revealing a marked downregulation of ion uptake mechanisms. The gill epithelium consists of thick, apical pillar cell flanges, the perikarya of which are coupled to an intralamellar septum. These two cell types respectively exhibit extensive apical evaginations and deep membrane invaginations, both of which are associated with numerous mitochondria, characterizing an ion transporting epithelium. These changes in Na(+)/K(+)- and V-ATPase activities and in mRNA expression during salinity acclimation appear to underpin ion uptake and Cl(-) secretion by the palaemonid shrimp gill. PMID:21037069

  15. Molecular characterization of insulin-like androgenic gland hormone-binding protein gene from the oriental river prawn Macrobrachium nipponense and investigation of its transcriptional relationship with the insulin-like androgenic gland hormone gene.

    PubMed

    Li, Fajun; Bai, Hongkun; Xiong, Yiwei; Fu, Hongtuo; Jiang, Sufei; Jiang, Fengwei; Jin, Shubo; Sun, Shengming; Qiao, Hui; Zhang, Wenyi

    2015-05-15

    Insulin-like androgenic gland hormone-binding protein (IAGBP) has been investigated in crustaceans in vitro. However, the relationship between IAGBP and its putative binding protein partner insulin-like androgenic gland hormone (IAG) has not been studied at the transcriptional level in vivo. In the current study, we cloned the full-length cDNA of IAGBP from the oriental river prawn Macrobrachium nipponense (Mn-IAGBP) and investigated the transcriptional patterns of Mn-IAGBP and the M. nipponense IAG gene (Mn-IAG) at different developmental stages and in different tissues. Mn-IAGBP mRNA was detected in all examined tissues from adult male prawns, with the highest transcriptional levels in the testis. Mn-IAG mRNA was detected in the androgenic gland and hepatopancreas. The genomic sequences of Mn-IAGBP and Mn-IAG were isolated by genome walking and two gene copies were found in both Mn-IAGBP and Mn-IAG. The relationship between Mn-IAGBP and Mn-IAG at the transcriptional level was studied by RNA interference. Injection of Mn-IAGBP double-stranded RNA (dsRNA) significantly reduced the transcription of Mn-IAG, while injection of Mn-IAG dsRNA significantly reduced the transcription of Mn-IAGBP in testis, muscle, androgenic gland, and hepatopancreas. These results demonstrate the involvement of the IAGBP gene in IAG signaling in M. nipponense. PMID:25558010

  16. 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.

  17. Identification of putative regulatory region of insulin-like androgenic gland hormone gene (IAG) in the prawn Macrobrachium nipponense and proteins that interact with IAG by using yeast two-hybrid system.

    PubMed

    Ma, Ke-Yi; Li, Jia-Le; Qiu, Gao-Feng

    2016-04-01

    Insulin-like androgenic gland hormone gene (IAG) is a sex regulator specifically expressed in male crustaceans, controlling the male sexual differentiation, spermatogenesis and reproductive strategy. Our previous study reported the cloning and characterization of the prawn Macrobrachium nipponense IAG (MnIAG). In this study, we further identified a 2214-bp MnIAG 5'-flanking region, and analyzed its transcription factor binding sites and transcriptional activity. The results showed that there were two potential promoter core sequences, three TATA boxes and one CAAT box existing in the MnIAG 5'-flanking region as well as many potential transcription factor binding sites, such as SRY, Sox-5, GATA-1, etc. Notably, the transcriptional activity was weak in this region, and a negative regulatory region was found in -604 to -231bp. In addition, we constructed M. nipponense yeast libraries and identified proteins interacting with the MnIAG protein by yeast two hybridization assay. The yeast two-hybrid screening yielded ten positive clones, of which five were annotated by NCBI database, namely heat shock protein 21, NADH dehydrogenase, zinc finger protein, beta-N-acetylglucosaminidase and a hypothetical protein. The identification of MnIAG putative regulatory region and proteins that interact with IAG will facilitate our understanding of the regulatory role of MnIAG and provide a foundation for deep insight into the prawn sex differentiation mechanism and signaling transduction pathways. PMID:26979275

  18. 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)

  19. 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.

  20. Studies on the ingestion characteristics of giant freshwater prawn, Chinese prawn and giant tiger prawn

    NASA Astrophysics Data System (ADS)

    Zang, Wei-Ling; Wang, Wei-Dong; Dai, Xi-Lin; Jiang, Min; Zhu, Zheng-Guo; Yang, Ming-Hui; Liu, Xian-Zhong; Xu, Gui-Rong; Ding, Fu-Jiang

    2000-12-01

    The ingestion of giant freshwater prawn, Chinese prawn and giant tiger prawn had continuity and the ingestion high peak occurred at night. Light and temperature had significant effects on the daily ingestion rate (DIR) of giant freshwater prawn Macrobrachium rosenbergii. Red light and blue light favorably induced favorable ingestion. In the adaptive range of temperature, the DIR increased with rising temperature and feeding frequency, but decreased with rising body weight.

  1. 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.

  2. Kinetic analysis of gill (Na⁺,K⁺)-ATPase activity in selected ontogenetic stages of the Amazon River shrimp, Macrobrachium amazonicum (Decapoda, Palaemonidae): interactions at ATP- and cation-binding sites.

    PubMed

    Leone, Francisco Assis; Masui, Douglas Chodi; de Souza Bezerra, Thais Milena; Garçon, Daniela Pereira; Valenti, Wagner Cotroni; Augusto, Alessandra Silva; McNamara, John Campbell

    2012-04-01

    We investigated modulation by ATP, Mg²⁺, Na⁺, K⁺ and NH₄⁺ and inhibition by ouabain of (Na⁺,K⁺)-ATPase activity in microsomal homogenates of whole zoeae I and decapodid III (formerly zoea IX) and whole-body and gill homogenates of juvenile and adult Amazon River shrimps, Macrobrachium amazonicum. (Na⁺,K⁺)-ATPase-specific activity was increased twofold in decapodid III compared to zoea I, juveniles and adults, suggesting an important role in this ontogenetic stage. The apparent affinity for ATP (K(M) = 0.09 ± 0.01 mmol L⁻¹) of the decapodid III (Na⁺,K⁺)-ATPase, about twofold greater than the other stages, further highlights this relevance. Modulation of (Na⁺,K⁺-ATPase activity by K⁺ also revealed a threefold greater affinity for K⁺ (K₀.₅ = 0.91 ± 0.04 mmol L⁻¹) in decapodid III than in other stages; NH₄⁺ had no modulatory effect. The affinity for Na⁺ (K₀.₅ = 13.2 ± 0.6 mmol L⁻¹) of zoea I (Na⁺,K⁺)-ATPase was fourfold less than other stages. Modulation by Na⁺, Mg²⁺ and NH₄⁺ obeyed cooperative kinetics, while K⁺ modulation exhibited Michaelis-Menten behavior. Rates of maximal Mg²⁺ stimulation of ouabain-insensitive ATPase activity differed in each ontogenetic stage, suggesting that Mg²⁺-stimulated ATPases other than (Na⁺,K⁺)-ATPase are present. Ouabain inhibition suggests that, among the various ATPase activities present in the different stages, Na⁺-ATPase may be involved in the ontogeny of osmoregulation in larval M. amazonicum. The NH₄⁺-stimulated, ouabain-insensitive ATPase activity seen in zoea I and decapodid III may reflect a stage-specific means of ammonia excretion since functional gills are absent in the early larval stages. PMID:22544049

  3. Cloning and characterization of the tiger shrimp lysozyme.

    PubMed

    Ye, Xing; Gao, Feng-Ying; Zheng, Qing-Mei; Bai, Jun-Jie; Wang, Huan; Lao, Hai-Hua; Jian, Qing

    2009-07-01

    Lysozymes are key proteins to invertebrates in the innate immune responses against bacterial infections. A lysozyme gene isolated from tiger shrimp, Penaeus monodon, was cloned, sequenced and characterized. The cDNA consists of a signal peptide of 18 amino acids and a mature peptide of 140 amino acids. The lysozyme is presumed to be a chicken-type lysozyme for it possesses two catalytic sites and eight cysteine residues which are highly conserved across species of chicken-type lysozymes. The lysozyme cDNAs of Penaeus semisulcatus, Litopenaeus vannamei, Macrobrachium nipponense and Macrobrachium rosenbergii were also cloned. High similarities existed among shrimp and prawn lysozymes but phylogenetic relationship of shrimps and prawns based on lysozyme molecules did not quite consistent with traditional taxonomic classification. High mRNA expression was detected in hepatopancreas, haemocytes and gill of tiger shrimp. Recombinant lysozyme exhibited potent lytic activities against fish pathogens providing evidence of the involvement of lysozyme in shrimp immunity. PMID:18618291

  4. Fish TRIM39 regulates cell cycle progression and exerts its antiviral function against iridovirus and nodavirus.

    PubMed

    Wang, Wei; Huang, Youhua; Yu, Yepin; Yang, Ying; Xu, Meng; Chen, Xiuli; Ni, Songwei; Qin, Qiwei; Huang, Xiaohong

    2016-03-01

    The tripartite motif (TRIM)-containing proteins exert important immune regulatory roles through regulating different signaling pathways in response to different stimuli. TRIM39, a member of the TRIM family, is a RING domain-containing E3 ubiquitin ligase which could regulate cell cycle progression and apoptosis. However, the antiviral activity of TRIM39 is not explored. Here, a TRIM39 homolog from grouper, Epinephelus coioides (EcTRIM39) was cloned, and its effects on cell cycle progression and fish virus replication were investigated. The full-length EcTRIM39 cDNA was composed of 2535 bp and encoded a polypeptide of 543 amino acids with 70% identity with TRIM39 homologs from bicolor damselfish. Amino acid alignment analysis indicated that EcTRIM39 contained a RING finger, B-box and SPRY domain. Expression profile analysis revealed that EcTRIM39 was abundant in intestine, spleen and skin. Upon different stimuli in vivo, the EcTRIM39 transcript was obviously up-regulated after challenging with Singapore grouper iridovirus (SGIV), and polyinosinic-polycytidylic acid (poly I:C). Using fluorescence microscopy, we found that EcTRIM39 localized in the cytoplasm and formed aggregates in grouper spleen (GS) cells. The ectopic expression of EcTRIM39 in vitro affected the cell cycle progression via mediating G1/S transition. Moreover, the RING domain was essential for its accurate localization and effect on cell cycle. In addition, overexpression of EcTRIM39 significantly inhibited viral gene transcription of SGIV and red-spotted grouper nervous necrosis virus (RGNNV) in vitro, and the mutant of RING exerted the opposite effect. Together, our results demonstrated that fish TRIM39 not only regulated the cell cycle progression, but also acted as an important regulator of fish innate immune response against viruses. PMID:26784918

  5. Coastal Aquaculture Development in Bangladesh: Unsustainable and Sustainable Experiences

    NASA Astrophysics Data System (ADS)

    Azad, A. Kalam; Jensen, Kathe R.; Lin, C. Kwei

    2009-10-01

    Coastal aquaculture in Bangladesh consists mainly of two shrimp species ( Penaeus monodon and Macrobrachium rosenbergii). Currently, there are about 16,237 marine shrimp ( P. monodon) farms covering 148,093 ha and 36,109 fresh water shrimp ( M. rosenbergii) farms covering 17,638 ha coastal area. More than 0.7 million people are employed in the farmed shrimp sector and in 2005-2006 the export value of shrimp was 403.5 million USD. Thus, coastal aquaculture contributes significantly to rural employment and economy but this is overshadowed by negative social and ecological impacts. This article reviews the key issues, constraints and opportunities of sustainable shrimp farming. In addition we present the results of two case studies from southwestern coastal areas where shrimp farming originated and central coastal areas where shrimp farming, especially M. rosenbergii, began in recent years. Lessons learned from the review and case studies are considered in the context of recommendations to encompass a socially equitable and ecologically sound coastal aquaculture.

  6. The invasion of five alien species in the Delta do Parnaíba Environmental Protection Area, Northeastern Brazil.

    PubMed

    Loebmann, Daniel; Mai, Ana Cecília G; Lee, James T

    2010-09-01

    Marine biological invasions have been regarded as one of the major causes of native biodiversity loss, with shipping and aquaculture being the leading contributors for the introductions of alien species in aquatic ecosystems. In the present study, five aquatic alien species (one mollusk, three crustaceans and one fish species) were detected during dives, shore searches and from the fisheries on the coast of the Delta do Parnaíba Environmental Protection Area, in the States of Piauí and Maranhão, Northeastern Brazil. The species were the bicolor purse-oyster Isognomon bicolor, the whiteleg shrimp Litopenaeus vannamei, the giant river prawn Macrobrachium rosenbergii, the Indo-Pacific swimming crab Charybdis hellerii and, the muzzled blenny Omobranchus punctatus. Ballast water (I. bicolor, C. hellerii, and O. punctatus) and aquaculture activities (L. vannamei and M. rosenbergii) in adjacent areas are the most likely vectors of introduction. All exotic species found have potential impact risks to the environment because they are able to compete against native species for resources (food and habitat). Isognomon bicolor share the same habitat and food items with the native bivalve species of mussels and barnacles. Litopenaeus vannamei share the same habitat and food items with the native penaeids such as the pinkspot shrimp Farfantepenaeus brasiliensis, the Southern brown shrimp Farfantepenaeus subtilis, and the Southern white shrimp Litopenaeus schmitti, and in the past few years L. vannamei was responsible for a viral epidemics in the cultivation tanks that could be transmitted to native penaeid shrimps. Charybdis hellerii is also able to cause impacts on the local fisheries as the species can decrease the populations of native portunid crabs which are commercialized in the studied region. Macrobrachium rosenbergii may be sharing natural resources with the Amazon River prawn Macrobrachium amazonicum. Omobranchus punctatus shares habit with the native redlip blenny Ophioblennius atlanticus and other fishes, such as the frillfin goby Bathigobius soporator Some immediate remedial measures to prevent further introductions from ballast water and shrimp farm ponds should be: (i) to prevent the release of ballast water by ship/vessels in the region; (ii) to reroute all effluent waters from shrimp rearing facilities through an underground or above-ground dry well; (iii) to install adequate sand and gravel filter which will allow passage of water but not livestock; (iv) outdoor shrimp pounds located on floodable land should be diked, and; (v) to promote environmental awareness of those directly involved with ballast water (crews of ship/vessels) and shrimp farms in the region. PMID:20737846

  7. 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 17β-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

  8. The occurrence of semicarbazide in the meat and shell of Bangladeshi fresh-water shrimp.

    PubMed

    McCracken, Robert; Hanna, Bob; Ennis, David; Cantley, Lynne; Faulkner, Dermot; Kennedy, D Glenn

    2013-02-15

    There is evidence that semicarbazide (SEM), a marker for the banned nitrofuran nitrofurazone, can arise from other, unrelated sources. Recently, Belgium rejected 54 consignments of Bangladeshi freshwater shrimp (Macrobrachium rosenbergii), following a laboratory decision to test meat and exoskeleton combined. To study the possible natural occurrence of SEM in wild shrimp, samples were collected and analysed from 29 sites across Bangladesh. SEM (<1.0 ?g/kg) was detected in ?65% of meat samples. However, SEM concentrations were approximately 100 times higher in the exoskeleton, and were unrelated to sampling location, strongly suggesting natural occurrence. In meat, most SEM was surface-associated. When the shrimp was shelled, some of the epidermal layer (which synthesises new exoskeleton) remained with the shell and some remained with the meat--leading to differing levels of natural SEM on the shrimp surface. This has implications for the use of SEM and the analytical strategy used to control nitrofuran use. PMID:23194563

  9. Effects of temperature and dissolved oxygen content on oxygen consumption rate of Chinese prawn, giant tiger prawn and giant freshwater prawn

    NASA Astrophysics Data System (ADS)

    Dai, Xi-Lin; Zang, Wei-Ling; Wang, Wei-Dong; Shi, Yong-Hai; Liu, Wen-Cui; Xu, Gui-Rong; Li, Shi-Hua

    1999-06-01

    Temperature and the dissolved oxygen content affect the oxygen consumption of juveniles of Chinese prawn ( Penaeus chinensis), giant tiger prawn ( P. monodon) and giant freshwater prawn ( Macrobrachium rosenbergii). There is good correlation between the oxygen consumption rate ( V, mg/g·h) of the above three prawn species and the water temperature, and dissolved oxygen. In the range of test temperature, V increased with water temperature and dissolved oxygen content. The V of the above three prawn species increased 0.085 mg/g·h, 0.093 mg/g·h and 0.08 mg/g·h respectively with each °C of rising temperature. The comatose point and stifling point of the juveniles rose obviously at unsuitable temperature.

  10. 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, Jarosław; 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, Soňa; Vera, Manuel; Villanova, Gabriela V; Voglmayr, Hermann; Vyskočilová, 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

  11. Deep parallel sequencing reveals conserved and novel miRNAs in gill and hepatopancreas of giant freshwater prawn.

    PubMed

    Tan, Tian Tian; Chen, Maoshan; Harikrishna, Jennifer Ann; Khairuddin, Norliana; Mohd Shamsudin, Maizatul Izzah; Zhang, Guojie; Bhassu, Subha

    2013-10-01

    MicroRNAs (miRNAs) are ~20-22 nucleotides, non protein-coding RNA regulatory genes that post-transcriptionally regulate many protein-coding genes, influencing critical biological and metabolic processes. While the number of known microRNA is increasing, there is currently no published data for miRNA from giant freshwater prawns, Macrobrachium rosenbergii (M. rosenbergii), a commercially cultured and economically important food species. In this study, we identified novel miRNAs in the gill and hepatopancreas of M. rosenbergii. Through a deep parallel sequencing analysis and an in silico data analysis approach, 327 miRNA families were identified from small RNA libraries with reference to both the de novo transcriptome of M. rosenbergii obtained from RNA-Seq and to miRBase (Release 18.0, November 2012). Based on the identified mature miRNA and recovered precursor sequences that form appropriate hairpin structures, three conserved miRNA (miR125, miR750, miR993) and 27 novel miRNA candidates encoding messenger-like non-coding RNA were identified. miR-125, miR-750, G-m0002/H-m0009, G-m0005, G-m0008/H-m0016, G-m0011/H-m0027 and G-m0015 were selected for experimental validation with stem-loop quantitative RT-PCR and were found to be coherent with the expression profile of deep sequencing data as evaluated with Pearson's correlation coefficient (r = 0.835178 for miRNA in gill, r = 0.724131 for miRNA in hepatopancreas). Using a combinatorial approach of pathway enrichment analysis and inverse expression relationship of miRNA and mRNA, four co-expressed novel miRNA candidates (G-m0005, G-m0008/H-m0016, G-m0011/H-m0027, and G-m0015) were found to be associated with energy metabolism. In addition, the expression of the three novel miRNA candidates (G-m0005, G-m0008/H-m0016, and G-m0011/H-m0027) were also found to be significantly reduced at 9 and 24 h post infection in M. rosenbergii challenged with infectious hypodermal and hematopoietic necrosis virus, suggesting a functional role of these miRNAs in crustacean immune defense. PMID:23816854

  12. Two host microRNAs influence WSSV replication via STAT gene regulation.

    PubMed

    Huang, Ying; Wang, Wen; Ren, Qian

    2016-01-01

    MicroRNAs (miRNAs) have important roles in post-transcriptional regulation of gene expression. During viral infection, viruses utilize hosts to enhance their replication by altering cellular miRNAs. The Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway plays crucial roles in the antiviral responses. In this study, two miRNAs (miR-9041 and miR-9850) from Macrobrachium rosenbergii were found to promote white spot syndrome virus (WSSV) replication. The up-regulation of miR-9041 or miR-9850 suppresses STAT expression in the gills of M. rosenbergii, which subsequently down-regulates the expression of its downstream dynamin (Dnm) genes: Dnm1, Dnm2, and Dnm3. Knockdown of miR-9041 and miR-9850 restricts WSSV replication by up-regulating STAT and Dnm gene expression. The silencing of STAT, Dnm1, Dnm2, or Dnm3 led to an increase of the number of WSSV copies in shrimp. The injection of recombinant Dnm1, Dnm2, or Dnm3 proteins could inhibit WSSV replication in vivo. Overall, our research indicates the roles of host miRNAs in the enhancement of WSSV replication by regulating the host JAK/STAT pathway. PMID:27029712

  13. 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

  14. Two host microRNAs influence WSSV replication via STAT gene regulation

    PubMed Central

    Huang, Ying; Wang, Wen; Ren, Qian

    2016-01-01

    MicroRNAs (miRNAs) have important roles in post-transcriptional regulation of gene expression. During viral infection, viruses utilize hosts to enhance their replication by altering cellular miRNAs. The Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway plays crucial roles in the antiviral responses. In this study, two miRNAs (miR-9041 and miR-9850) from Macrobrachium rosenbergii were found to promote white spot syndrome virus (WSSV) replication. The up-regulation of miR-9041 or miR-9850 suppresses STAT expression in the gills of M. rosenbergii, which subsequently down-regulates the expression of its downstream dynamin (Dnm) genes: Dnm1, Dnm2, and Dnm3. Knockdown of miR-9041 and miR-9850 restricts WSSV replication by up-regulating STAT and Dnm gene expression. The silencing of STAT, Dnm1, Dnm2, or Dnm3 led to an increase of the number of WSSV copies in shrimp. The injection of recombinant Dnm1, Dnm2, or Dnm3 proteins could inhibit WSSV replication in vivo. Overall, our research indicates the roles of host miRNAs in the enhancement of WSSV replication by regulating the host JAK/STAT pathway. PMID:27029712

  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. Haemato-biochemical disorders associated with nodavirus like-agent in adult leaping mullet Liza saliens (Risso, 1810) in the Caspian Sea.

    PubMed

    Ghiasi, Maryam; Binaii, Mohammad; Ghasemi, Mohdes; Fazli, Hasan; Zorriehzahra, Mohamad Jalil

    2016-03-01

    Betanoda virus is an emerging problem in several marine fish species in various geographic areas all over the world. In recent years, mullets stock of the Caspian Sea decreased dramatically and a betanoda virus like-agent was introduced as the cause of mullet's mortality. The main objective of the present study is to compare hemato-biochemical parameters in healthy and infected sharpnose mullets (Liza saliens) to betanoda virus like-agent. The adult sharpnose mullets (34 clinically affected fish + 34 apparently healthy fish) were captured from the southeast the Caspian Sea from 2012 to 2013. All of the captured fish were 4 or 5 years old. The main clinical sings of infected fish were lethargy, severe abdominal distention, abnormal swimming and hyperinflation of swim bladder. The results showed that the weight and total length of infected fish were significantly lower than the healthy fish. The results of histopatological evaluation and indirect florescent antibody test were confirmed the presence of a betanoda virus-like agent in infected fish. The diseased fish were severely anemic (hypochromic macrocytic anemia) and had a serious haemopoietic disorders. The anemia associated with a sever leukopenia, a significant rise of neutrophils and immature neutrophils and a significant decries of lymphocytes percentage. Total protein, albumin and total immunoglobulin levels were significantly reduced in the serum of infected fish, while the activities of aspartate aminotransferase and alanine aminotransferase significantly increased when compared to the healthy fish. These results suggested that the feeding of the infected fish to betanoda virus like-agent were disturbed and it could be cause haemato-biochemical disorder and mortality of the fish in Iranian water of the Caspian Sea. PMID:26925439

  17. Molecular importance of prawn large heat shock proteins 60, 70 and 90.

    PubMed

    Chaurasia, Mukesh Kumar; Nizam, Faizal; Ravichandran, Gayathri; Arasu, Mariadhas Valan; Al-Dhabi, Naif Abdullah; Arshad, Aziz; Elumalai, Preetham; Arockiaraj, Jesu

    2016-01-01

    Considering the importance of heat shock proteins (HSPs) in the innate immune system of prawn, a comparative molecular approach was proposed to study the crustacean large HSPs 60, 70 and 90. Three different large HSPs were identified from freshwater prawn Macrobrachium rosenbergii (Mr) cDNA library during screening. The structural and functional characteristic features of HSPs were studied using various bioinformatics tools. Also, their gene expression and mRNA regulation upon various pathogenic infections was studied by relative quantification using 2(-ΔΔCT) method. MrHSP60 contains a long chaperonin 60 domain at 46-547 which carries a chaperonin 60 signature motif between 427 and 438, whereas MrHSP70 contains a long HSP70 domain at 21-624 and MrHSP90 carries a HSP90 domain at 188-719. The two dimensional analysis showed that MrHSP60 contains more amino acids (52%) in helices, whereas MrHSP70 (40.6%) and MrHSP90 (51.8%) carried more residues in coils. Gene expression results showed significant (P < 0.05) expression of MrHSP60, 70 and 90 in haemocyte, gill and hepatopancreas, respectively. Further, the expression level was up-regulated upon bacterial (Aeromonas hydrophilla and Vibrio harveyi) and viral [white spot syndrome virus (WSSV) and M. rosenbergii nodo virus (MrNV)] infections during various time periods. The gene expression results exhibited the potential involvement of these three HSPs in the immune system of prawn. The study indicated the potentiality of these molecules, thereby protecting cells against pathogens as well as severe cellular and environmental stresses in crustaceans. PMID:26631804

  18. Expression of the male reproduction-related gene in spermatic ducts of the blue swimming crab, Portunus pelagicus, and transfer of modified protein to the sperm acrosome.

    PubMed

    Sroyraya, Morakot; Hanna, Peter J; Changklungmoa, Narin; Senarai, Thanyaporn; Siangcham, Tanapan; Tinikul, Yotsawan; Sobhon, Prasert

    2013-01-01

    Expression of a sex-specific gene in Macrobrachium rosenbergii (Mr-Mrr), encoding a male reproduction-related (Mrr) protein, has been identified in the spermatic ducts (SDs) and postulated to be involved in sperm maturation processes. M. rosenbergii is the only decapod that the expression and fate of the Mrr protein has been studied. To determine that this protein was conserved in decapods, we firstly used cloning techniques to identify the Mrr gene in two crabs, Portunus pelagicus (Pp-Mrr) and Scylla serrata (Ss-Mrr). We then investigated expression of Pp-Mrr by in situ hybridization, and immunolocalization, as well as phosphorylation and glycosylation modifications, and the fate of the protein in the male reproductive tract. Pp-Mrr was shown to have 632 nucleotides, and a deduced protein of 110 amino acids, with an unmodified molecular weight of 11.79 kDa and a mature protein with molecular weight of 9.16 kDa. In situ hybridization showed that Pp-Mrr is expressed in the epithelium of the proximal, middle, distal SDs, and ejaculatory ducts. In Western blotting, proteins of 10.9 and 17.2 kDa from SDs were all positive using anti-Mrr, antiphosphoserine/threonine, and antiphosphotyrosine. PAS staining showed they were also glycosylated. Immunolocalization studies showed Pp-Mrr in the SD epithelium, lumen, and on the acrosomes of spermatozoa. Immunofluorescence staining indicated the acrosome of spermatozoa contained the Mrr protein, which is phosphorylated with serine/threonine and tyrosine, and also glycosylated. The Mrr is likely to be involved in acrosomal activation during fertilization of eggs. PMID:23108973

  19. Identification and Characterization of an Insulin-Like Receptor Involved in Crustacean Reproduction.

    PubMed

    Sharabi, O; Manor, R; Weil, S; Aflalo, E D; Lezer, Y; Levy, T; Aizen, J; Ventura, T; Mather, P B; Khalaila, I; Sagi, A

    2016-02-01

    Sexual differentiation and maintenance of masculinity in crustaceans has been suggested as being regulated by a single androgenic gland (AG) insulin-like peptide (IAG). However, downstream elements involved in the signaling cascade remain unknown. Here we identified and characterized a gene encoding an insulin-like receptor in the prawn Macrobrachium rosenbergii (Mr-IR), the first such gene detected in a decapod crustacean. In mining for IRs and other insulin signaling-related genes, we constructed a comprehensive M. rosenbergii transcriptomic library from multiple sources. In parallel we sequenced the complete Mr-IR cDNA, confirmed in the wide transcriptomic library. Mr-IR expression was detected in most tissues in both males and females, including the AG and gonads. To study Mr-IR function, we performed long-term RNA interference (RNAi) silencing in young male prawns. Although having no effect on growth, Mr-IR silencing advanced the appearance of a male-specific secondary trait. The most noted effects of Mr-IR silencing were hypertrophy of the AG and the associated increased production of Mr-IAG, with an unusual abundance of immature sperm cells being seen in the distal sperm duct. A ligand blot assay using de novo recombinant Mr-IAG confirmed the existence of a ligand-receptor interaction. Whereas these results suggest a role for Mr-IR in the regulation of the AG, we did not see any sexual shift after silencing of Mr-IR, as occurred when the ligand-encoding Mr-IAG gene was silenced. This suggests that sexual differentiation in crustaceans involve more than a single Mr-IAG receptor, emphasizing the complexity of sexual differentiation and maintenance. PMID:26677879

  20. Production of recombinant insulin-like androgenic gland hormones from three decapod species: In vitro testicular phosphorylation and activation of a newly identified tyrosine kinase receptor from the Eastern spiny lobster, Sagmariasus verreauxi.

    PubMed

    Aizen, Joseph; Chandler, Jennifer C; Fitzgibbon, Quinn P; Sagi, Amir; Battaglene, Stephen C; Elizur, Abigail; Ventura, Tomer

    2016-04-01

    In crustaceans the insulin-like androgenic gland hormone (IAG) is responsible for male sexual differentiation. To date, the biochemical pathways through which IAG exerts its effects are poorly understood and could be elucidated through the production of a functional recombinant IAG (rIAG). We have successfully expressed glycosylated, biologically active IAG using the Pichia pastoris yeast expression system. We co-expressed recombinant single-chain precursor molecules consisting of the B and A chains (the mature hormone) tethered by a flexible linker, producing rIAGs of the following commercially important species: Eastern spiny lobster Sagmariasus verreauxi (Sv), redclaw crayfish Cherax quadricarinatus (Cq) and giant freshwater prawn Macrobrachium rosenbergii (Mr). We then tested the biological activity of each, through the ability to increase phosphorylation in the testis; both Sv and Cq rIAGs significantly elevated phosphorylation specific to their species, and in a dose-dependent manner. Mr rIAG was tested on Macrobrachium australiense (Ma), eliciting a similar response. Moreover, using bioinformatics analyses of the de novo assembled spiny lobster transcriptome, we identified a spiny lobster tyrosine kinase insulin receptor (Sv-TKIR). We validated this discovery with a receptor activation assay in COS-7 cells expressing Sv-TKIR, using a reporter SRE-LUC system designed for RTKs, with each of the rIAG proteins acting as the activation ligand. Using recombinant proteins, we aim to develop specific tools to control sexual development through the administration of IAG within the critical sexual differentiation time window. The biologically active rIAGs generated might facilitate commercially feasible solutions for the long sought techniques for sex-change induction and monosex population culture in crustaceans and shed new light on the physiological mode of action of IAG in crustaceans. PMID:26883686

  1. 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.

  2. 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

  3. 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

  4. Comparison of genetic characteristics and pathogenicity of Lactococcus garvieae isolated from aquatic animals in Taiwan.

    PubMed

    Tsai, Ming-An; Wang, Pei-Chyi; Liaw, Li-Ling; Yoshida, Terutoyo; Chen, Shih-Chu

    2012-12-01

    Seventy-six Taiwanese bacterial isolates including 74 from diseased, cultured, aquatic animals (54 grey mullet Mugil cephalus, 3 basket mullet Chelon alatus, 2 tilapia Oreochromis niloticus, 1 grouper Epinephelus coioides, 2 yellowfin seabream Acanthopagrus latus, 1 Borneo mullet Chelon macrolepis, 1 bullfrog Rana catesbeiana, 1 Japanese eel Anguilla japonica, and 9 giant freshwater prawns Macrobrachium rosenbergii), 1 wild-caught seafood species (squid muscle collected from a restaurant) and 1 human isolate (from a patient with a history of consuming raw squid in the previously mentioned restaurant), all collected between 1999 and 2006, were confirmed by PCR assay to be Lactococcus garvieae. The phenotypic characterization was determined by rabbit anti-KG+ and KG- serums, and 74 of the 76 Taiwanese strains displayed a KG- phenotype. The genetic characterization was investigated by pulsed-field gel electrophoresis (PFGE). Genomic DNA was digested with restriction endonucleases ApaI and SmaI and separated by PFGE. Ten different L. garvieae pulsotypes were identified. Predominant pulsotypes A1a/S1a were obtained from >96% of strains (52 of 54) from grey mullet, demonstrating a clonal dissemination of L. garvieae in grey mullet in Taiwan. In experimental challenges with grey mullet and tilapia, L. garvieae pulsotypes A1/S1 and A11/S11 showed higher virulence compared with other pulsotypes. PMID:23209077

  5. Utilization of warm well water, eastern Washington state

    NASA Astrophysics Data System (ADS)

    1982-03-01

    Utilizing the warm well water for a geothermal greenhouse heating system is highly economically feasible. This is based on using the 88 F water from Anderson Well No. 1 to heat greenhouses totaling approximately 10.6 acres. The additional investment of $640,000 above the cost for a conventional electric boiler system shows a rate of return of 48.3% on a 20 year life cycle analysis. The simple payback is 3 years. The 88 F well water is not warm enough for prawn (Macrobrachium rosenbergii) aquaculture, since water flow requirements are excessive to maintain the desired 80 F pond temperature. However, the water is warm enough to maintain a 60 F pond temperature for trout farming. Trout farming using the 88 F well water directly is probably not economically feasible due to high electrical pumping cost (34,626 per year) for the seven 1/2 acre ponds that could be heated. Trout farming using the 75 F effluent water from the 10.6 acre greenhouse to heat four 1/2 acre ponds may be economically feasible since the water booster pumping cost is low ($1189 per year).

  6. Seasonal abundance and composition of finfish and shellfish seeds inmangroves of Gangolli estuary, off south-west coast of India.

    PubMed

    Rajesh, K M; Thomas, Sujitha; Dineshbabu, A R; PrathibhaRohit; Sridhara, B; Nataraja, G D

    2015-11-01

    The composition and seasonal abundance of the fin and shellfish seeds in mangrove areas of Gangoli estuary were studied for one year from January to December, 2013. Monthly day time sampling was conducted with a rectangular dragnet of 75 m2 length having 5 mm mesh size. The mean monthly density was highest in January (Avg. of 12,996/haul) and lowest during May (Avg. of 859/haul). Thirty species of fish, four species of shrimps and two species of crabs belonging to 26 families were recorded during the study. Ambasis sp., Feneropenaeus indicus, Etroplus suratensis,Leognathus sp., Liza sp., Stolephorus vaganensis, Hemiramphus sp., Terapon sp. and Gerres sp. were the most abundant species/taxa. A total of 10 commercially important species comprising F. indicus, Etroplus suratensis, Liza sp., Gerres sp.,Siganus sp., Lutjanus russei, Macrobrachium rosenbergii, Lutjanus argentimaculatus, Sillago sp. and Penaeus monodon constituted 8.5% (April, 2013) to 93.7% (October, 2013) with an average of 37.2% of the total fish and shellfish seeds. Higher Species diversity (H') and Species richness (D) was recorded during January. The result of the present investigation gives an insight of the abundance and seasonality of commercially important fish and shellfish seeds, which may facilitate the brackishwater fish farmers to exploitthe seeds more pragmatically for their culture practice. PMID:26688975

  7. Geothermal aquaculture project: Real Property Systems Inc. , Harney Basin, Oregon

    SciTech Connect

    Not Available

    1981-08-14

    Real Property Systems Inc., (RPS) owns two parcels in the vicinity of Harney Lake, Oregon. One parcel is 120 acres in size, the other is 200 acres. A study concludes that the 200 acre parcel has the greater potential for geothermal development. RPS is interested in an aquaculture operation that produces fresh water prawns, (Macrobrachium rosenbergii) for the market. To supply the heat necessary to maintain the ideal temperature of 82/sup 0/F desired for these prawns, a geothermal resource having a 150/sup 0/F temperature or higher, is needed. The best estimate is that 150/sup 0/F water can be found from a minimum 1090 feet depth to 2625 feet, with no absolute assurances that sufficient quantities of geothermal waters exist without drilling for the same. This study undertakes the preliminary determination of project economics so that a decision can be made whether or not to proceed with exploratory drilling. The study is based on 10 acres of ponds, with a peak requirement of 2500 gpm of 150/sup 0/F geothermal water.

  8. Expression of an Androgenic Gland-Specific Insulin-Like Peptide during the Course of Prawn Sexual and Morphotypic Differentiation

    PubMed Central

    Ventura, Tomer; Manor, Rivka; Aflalo, Eliahu D.; Weil, Simy; Khalaila, Isam; Rosen, Ohad; Sagi, Amir

    2011-01-01

    The crustacean male-specific androgenic gland (AG) regulates sexual differentiation. In the prawn Macrobrachium rosenbergii, silencing an AG-specific insulin-like encoding transcript (Mr-IAG) inhibited the development of male sexual characters, suggesting that Mr-IAG is a key androgenic hormone. We used recombinant pro-Mr-IAG peptide to generate antibodies that recognized the peptide in AG cells and extracts, as verified by mass spectrometry. We revealed the temporal expression pattern of Mr-IAG and studied its relevance to the timetable of sex differentiation processes in juveniles and after puberty. Mr-IAG was expressed from as early as 20 days after metamorphosis, prior to the appearance of external male sexual characters. Mr-IAG expression was lower in the less reproductively active orange-clawed males than in both the dominant blue-clawed males and the actively sneak mating small males. These results suggest a role for Mr-IAG both in the timing of male sexual differentiation and in regulating reproductive strategies. PMID:22363879

  9. Fishing for prawn larvae in Bangladesh: an important coastal livelihood causing negative effects on the environment.

    PubMed

    Ahmed, Nesar; Troell, Max

    2010-02-01

    Freshwater prawn (Macrobrachium rosenbergii) farming in Bangladesh has, to a large extent, been dependent on the supply of wild larvae. Although there are 81 freshwater prawn hatcheries in the country, a lack of technical knowledge, inadequate skilled manpower, and an insufficient supply of wild broods have limited hatchery production. Many thousands of coastal poor people, including women, are engaged in fishing for wild prawn larvae along the coastline during a few months each year. On average, 40% of the total yearly income for these people comes from prawn larvae fishing activity. However, indiscriminate fishing of wild larvae, with high levels of bycatch of juvenile fish and crustaceans, may impact negatively on production and biodiversity in coastal ecosystems. This concern has provoked the imposition of restrictions on larvae collection. The ban has, however, not been firmly enforced because of the limited availability of hatchery-raised larvae, the lack of an alternative livelihood for people involved in larvae fishing, and weak enforcement power. This article discusses the environmental and social consequences of prawn larvae fishing and concludes that, by increasing awareness among fry fishers, improving fishing techniques (reducing bycatch mortality), and improving the survival of fry in the market chain, a temporal ban may be a prudent measure when considering the potential negative impacts of bycatch. However, it also suggests that more research is needed to find out about the impact of larvae fishing on nontarget organisms and on the populations of targeted species. PMID:20496649

  10. Effect of legume seed extracts on the inhibition of proteolytic activity and muscle degradation of fresh water prawn (Macrobrachiumrosenbergii).

    PubMed

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

    2011-12-01

    Trypsin inhibitors in the extracts from soybean (Glycine max), adzuki bean (Vigna angularis), bambara groundnut (Vigna subterranea) and red kidney bean (Phaseoulus vulgaris) varied in amount and molecular weight. The soybean extract had the highest level of trypsin inhibitor with molecular weight (MW) of 21kDa, followed by bambara groundnut extract possessing trypsin inhibitor with MW of 15kDa. Both extracts showed a more effective inhibition towards crude protease extract (CE) from the hepatopancreas of fresh water prawn (Macrobrachium rosenbergii) than the extracts from adzuki and red kidney beans. Activity staining also reconfirmed the higher inhibitory activity on CE from hepatopancreas by the extracts from both soybean and bambara groundnut. The extracts from all seeds were able to inhibit the degradation of fresh water prawn meat containing CE in a concentration dependent manner. Based on inhibitor study, the extracts from soybean and bambara groundnut can be a potential aid to suppress the muscle softening of fresh water prawn, mediated by trypsin-like proteases released from hepatopancreas, during extended iced storage. PMID:25212342

  11. Treatment efficiency of effluent prawn culture by wetland with floating aquatic macrophytes arranged in series.

    PubMed

    Henares, M N P; Camargo, A F M

    2014-11-01

    The efficiency of a series of wetland colonized with Eichhornia crassipes and Salvinia molesta to treat the effluent of a giant river prawn (Macrobrachium rosenbergii) broodstock pond was evaluated in this study. The experimental design was completely randomized and was performed in 9 rectangular tanks (1.6 m3) with three treatments (constructed wetlands) and three replicates. The treatment types included: a wetland colonized with E. crassipes and S. molesta (EcSm) arranged sequentially, a wetland with E. crassipes only (Ec) and a wetland with S. molesta only (Sm). The means of suspended particulate material (SPM), total inorganic nitrogen (TIN), total Kjeldahl nitrogen (TKN), P-orthophosphate (PO4-P) and total phosphorus (TP) of the treated effluents were compared using ANOVA followed by Tukey's test (P<0.05). The effluent treated in Ec and EcSm wetlands exhibited lower SPM concentrations. The Ec wetland reduced TIN, TKN, PO4-P and TP by 46.0, 43.7, 44.4 and 43.6%, respectively. In the EcSm wetland, the reduction of TIN (23.0%), TKN (33.7%) and PO4-P (26.7%) was similar to the Sm wetland (19.8% TIN, 30.9% TKN and 23.8% PO4-P). The Ec wetland was more efficient in treating pond effluent due likely to the higher root surface of E. crassipes, which forms an extensive area favorable to retention and adsorption of debris and absorption of nutrients. PMID:25627602

  12. Enhancement of trypsin-like enzymes by A23187 ionophore is crucial for sperm penetration through the egg vestment of the giant freshwater prawn.

    PubMed

    Watthammawut, Atthaboon; Somrit, Monsicha; Asuvapongpatana, Somluk; Weerachatyanukul, Wattana

    2015-12-01

    We report the presence of trypsin-like enzymes preferring Boc-QAR-MCA substrate in sperm collected from different portions of male reproductive tracts of the giant freshwater prawn, Macrobrachium rosenbergii and compare enzyme activities before and after an A23187 calcium ionophore treatment. Fluorogenic enzyme assays revealed that testicular sperm lysates showed high trypsin-like enzyme activity but the activity was relatively low in vas deferens sperm lysates as well as in the live sperm. Upon sperm treatment with A23187, trypsin-like activity was greatly enhanced in distal vas deferens sperm. Substrate- and inhibitor-based localization studies indicated that the sperm trypsin-like enzymes were not of a soluble type but were rather of a membrane-borne type, localized at the anterior spike and upper part of the main body. Notable structural changes were also evident in A23187-induced sperm including extensive ruffling of the sperm membrane structure at the base of the main body thereby supporting the acrosome reaction response in this species. We further proved by substrate inhibition assays that the enhanced trypsin-like enzyme activity participates in sperm penetration through the vitelline envelope, a novel sperm-egg penetration mechanism that is unique in this species. PMID:26123168

  13. 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

  14. Utilization of warm well water, eastern Washington State

    SciTech Connect

    Not Available

    1982-03-01

    Utilizing the warm well water for a geothermal greenhouse heating system is highly economically feasible. This is based on using the 88/sup 0/F water from Anderson Well No. 1 to heat greenhouses totaling approximately 10.6 acres. The additional investment of $640,000 above the cost for a conventional electric boiler system shows a rate of return of 48.3% on a 20 year life cycle analysis. The simple payback is 3 years. The 88/sup 0/F well water is not warm enough for prawn (Macrobrachium rosenbergii) aquaculture, since water flow requirements are excessive to maintain the desired 80/sup 0/F pond temperature. However, the water is warm enough to maintain a 60/sup 0/F pond temperature for trout farming. Trout farming using the 88/sup 0/F well water directly is probably not economically feasible due to high electrical pumping cost (34,626 per year) for the seven 1/2 acre ponds that could be heated. Trout farming using the 75/sup 0/F effluent water from the 10.6 acre greenhouse to heat four 1/2 acre ponds may be economically feasible since the water booster pumping cost is low $1189 per year.

  15. Effect of Shrimp Chitin and Shrimp Chitin Hydrolysate on the Freeze-Induced Denaturation, and on the Amount of Unfreezable Water of Wanieso Lizardfish Myofibrillar Protein

    NASA Astrophysics Data System (ADS)

    Somjit, Kingduean; Kongpun, Orawan; Osatomi, Kiyoshi; Hara, Kenji; Nozaki, Yukinori

    In view of potential utilization of shrimp waste, shrimp chitin (SC) and shrimp chitin hydrolysate (SCH) were prepared from 3 kinds of shrimp species, namely: black tiger shrimp Penaeus monodon, endeavour shrimp Metapenaeus endeavouri and giant freshwater shrimp Macrobrachium rosenbergii. The effects of 5% SC and SCH (dry weight) on the state of water and on the denaturation of wanieso lizardfish Saurida wanieso myofibrillar protein (Mf) were evaluated based on changes in Mf Ca-ATPase activity and the amount of unfreezable water during frozen storage. Each effect was compared with those of Mf without additives (control) and Mf with glucose. The changes in Ca-ATPase activity of control and Mf with SC during frozen storage were exhibited biphasic pattern while those of SCH and glucose exhibited monophasic pattern. The amount of unfreezable water of Mf with SC was lower than that of control while those of Mf with SCH and glucose were higher than that of control. Present findings suggested that the preventive effect of SCH on freeze-induced denaturation of Mf is caused by the stabilizing the hydrated water molecule surrounding the Mf.

  16. 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.

  17. 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.

  18. 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

  19. Poly--hydroxybutyrate content and dose of the bacterial carrier for Artemia enrichment determine the performance of giant freshwater prawn larvae.

    PubMed

    Thai, Truong Quoc; Wille, Mathieu; Garcia-Gonzalez, Linsey; Sorgeloos, Patrick; Bossier, Peter; De Schryver, Peter

    2014-06-01

    The beneficial effects of poly-?-hydroxybutyrate (PHB) for aquaculture animals have been shown in several studies. The strategy of applying PHB contained in a bacterial carrier has, however, hardly been considered. The effect of administering PHB-accumulated Alcaligenes eutrophus H16 containing 10 or 80% PHB on dry weight, named A10 and A80, respectively, through the live feed Artemia was investigated on the culture performance of larvae of the giant freshwater prawn (Macrobrachium rosenbergii). Feeding larvae with Artemia nauplii enriched in a medium containing 100 and 1,000mgL(-1) A80 significantly increased the survival with about 15% and the development of the larvae with a larval stage index of about 1 as compared to feeding non-enriched Artemia. The survival of the larvae also significantly increased with about 35% in case of a challenge with Vibrio harveyi. The efficiency of these treatments was equal to a control treatment of Artemia enriched in an 800 mgL(-1) PHB powder suspension, while Artemia enriched in 10mgL(-1) A80, 100mgL(-1) A10, and 1,000mgL(-1) A10 did not bring similar effects. From our results, it can be concluded that PHB supplemented in a bacterial carrier (i.e., amorphous PHB) can increase the larviculture efficiency of giant freshwater prawn similar to supplementation of PHB in powdered form (i.e., crystalline PHB). When the level of PHB in the bacterial carrier is high, similar beneficial effects can be achieved as crystalline PHB, but at a lower live food enrichment concentration expressed on PHB basis. PMID:24615382

  20. 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

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

    PubMed Central

    Jezzini, Sami H.; Reyes-Coln, Dalyns; Sosa, Mara 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

  2. Characterization of a prawn OA/TA receptor in Xenopus oocytes suggests functional selectivity between octopamine and tyramine.

    PubMed

    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

  3. 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

  4. 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

  5. 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

  6. Real-time RT-PCR detection of betanodavirus in naturally and experimentally infected fish from Spain.

    PubMed

    Hodneland, K; García, R; Balbuena, J A; Zarza, C; Fouz, B

    2011-03-01

    Infections with betanodavirus affect a wide range of wild and farmed fish species throughout the world, mostly from the marine environment. The aim of this work was to develop and validate real-time RT-PCR assays for sensitive and specific detection of nodavirus in diseased or carrier fish. The new detection assay was used to study the transmission and development of nodavirus infection in juvenile sea bass, Dicentrarchus labrax (L.), challenged by different routes, and also to screen for nodavirus in various farmed fish species. On average, the sensitivity was 10-100 times higher than a standard RT-PCR, and the assay was able to detect asymptomatic carrier fish that otherwise could have been classified as free of infection. Clinical signs of nodavirus infection were reproduced in fish infected following bath exposure or intramuscular injection, demonstrating horizontal transmission of the disease. Nodavirus was always detected in the brain of diseased fish but also in many recovered fish. The new assay enables us to confirm the presence of the virus at an early phase in the production cycle and may represent a useful tool to prevent or slow down the spread of nodavirus to new locations. PMID:21306586

  7. Pigment granule translocation in red ovarian chromatophores from the palaemonid shrimp Macrobrachium olfersi (Weigmann, 1836): functional roles for the cytoskeleton and its molecular motors.

    PubMed

    Milograna, Sarah Ribeiro; Ribeiro, Márcia Regina; Baqui, Munira Muhammad Abdel; McNamara, John Campbell

    2014-12-01

    The binding of red pigment concentrating hormone (RPCH) to membrane receptors in crustacean chromatophores triggers Ca²⁺/cGMP signaling cascades that activate cytoskeletal motors, driving pigment granule translocation. We investigate the distributions of microfilaments and microtubules and their associated molecular motors, myosin and dynein, by confocal and transmission electron microscopy, evaluating a functional role for the cytoskeleton in pigment translocation using inhibitors of polymer turnover and motor activity in vitro. Microtubules occupy the chromatophore cell extensions whether the pigment granules are aggregated or dispersed. The inhibition of microtubule turnover by taxol induces pigment aggregation and inhibits re-dispersion. Phalloidin-FITC actin labeling, together with tannic acid fixation and ultrastructural analysis, reveals that microfilaments form networks associated with the pigment granules. Actin polymerization induced by jasplaquinolide strongly inhibits RPCH-induced aggregation, causes spontaneous pigment dispersion, and inhibits pigment re-dispersion. Inhibition of actin polymerization by latrunculin-A completely impedes pigment aggregation and re-dispersion. Confocal immunocytochemistry shows that non-muscle myosin II (NMMII) co-localizes mainly with pigment granules while blebbistatin inhibition of NMMII strongly reduces the RPCH response, also inducing spontaneous pigment dispersion. Myosin II and dynein also co-localize with the pigment granules. Inhibition of dynein ATPase by erythro-9-(2-hydroxy-3-nonyl) adenine induces aggregation, inhibits RPCH-triggered aggregation, and inhibits re-dispersion. Granule aggregation and dispersion depend mainly on microfilament integrity although microtubules may be involved. Both cytoskeletal polymers are functional only when subunit turnover is active. Myosin and dynein may be the molecular motors that drive pigment aggregation. These mechanisms of granule translocation in crustacean chromatophores share various features with those of vertebrate pigment cells. PMID:25182860

  8. Differential distribution of V-type H(+)-ATPase and Na (+)/K (+)-ATPase in the branchial chamber of the palaemonid shrimp Macrobrachium amazonicum.

    PubMed

    Boudour-Boucheker, Nesrine; Boulo, Viviane; Charmantier-Daures, Mireille; Grousset, Evelyse; Anger, Klaus; Charmantier, Guy; Lorin-Nebel, Catherine

    2014-07-01

    V-H(+)-ATPase and Na(+)/K(+)-ATPase were localized in the gills and branchiostegites of M. amazonicum and the effects of salinity on the branchial chamber ultrastructure and on the localization of transporters were investigated. Gills present septal and pillar cells. In freshwater (FW), the apical surface of pillar cells is amplified by extensive evaginations associated with mitochondria. V-H(+)-ATPase immunofluorescence was localized in the membranes of the apical evaginations and in clustered subapical areas of pillar cells, suggesting labeling of intracellular vesicle membranes. Na(+)/K(+)-ATPase labeling was restricted to the septal cells. No difference in immunostaining was recorded for both proteins according to salinity (FW vs. 25 PSU). In the branchiostegite, both V-H(+)-ATPase and Na(+)/K(+)-ATPase immunofluorescence were localized in the same cells of the internal epithelium. Immunogold revealed that V-H(+)-ATPase was localized in apical evaginations and in electron-dense areas throughout the inner epithelium, while Na(+)/K(+)-ATPase occurred densely along the basal infoldings of the cytoplasmic membrane. Our results suggest that morphologically different cell types within the gill lamellae may also be functionally specialized. We propose that, in FW, pillar cells expressing V-H(+)-ATPase absorb ions (Cl(-), Na(+)) that are transported either directly to the hemolymph space or through a junctional complex to the septal cells, which may be responsible for active Na(+) delivery to the hemolymph through Na(+)/K(+)-ATPase. This suggests a functional link between septal and pillar cells in osmoregulation. When shrimps are transferred to FW, gill and branchiostegite epithelia undergo ultrastructural changes, most probably resulting from their involvement in osmoregulatory processes. PMID:24805036

  9. Modulation By K+ Plus NH4+ of Microsomal (Na+, K+)-ATPase Activity in Selected Ontogenetic Stages of the Diadromous River Shrimp Macrobrachium amazonicum (Decapoda, Palaemonidae)

    PubMed Central

    Leone, Francisco A.; Bezerra, Thais M. S.; Garçon, Daniela P.; Lucena, Malson N.; Pinto, Marcelo R.; Fontes, Carlos F. L.; McNamara, John C.

    2014-01-01

    We investigate the synergistic stimulation by K+ plus NH4+ of (Na+, K+)-ATPase activity in microsomal preparations of whole zoea I and decapodid III, and in juvenile and adult river shrimp gills. Modulation of (Na+, K+)-ATPase activity is ontogenetic stage-specific, and particularly distinct between juveniles and adults. Although both gill enzymes exhibit two different sites for K+ and NH4+ binding, in the juvenile enzyme, these two sites are equivalent: binding by both ions results in slightly stimulated activity compared to that of a single ionic species. In the adult enzyme, the sites are not equivalent: when one ion occupies its specific binding site, (Na+, K+)-ATPase activity is stimulated synergistically by ≈50% on binding of the complementary ion. Immunolocalization reveals the enzyme to be distributed predominantly throughout the intralamellar septum in the gill lamellae of juveniles and adults. Western blot analyses demonstrate a single immunoreactive band, suggesting a single (Na+, K+)-ATPase α-subunit isoform that is distributed into different density membrane fractions, independently of ontogenetic stage. We propose a model for the modulation by K+ and NH4+ of gill (Na+, K+)-ATPase activity. These findings suggest that the gill enzyme may be regulated by NH4+ during ontogenetic development in M. amazonicum. PMID:24586919

  10. Molecular profiles and pathogen-induced transcriptional responses of prawn B cell lymphoma-2 related ovarian killer protein (BOK).

    PubMed

    Chaurasia, Mukesh Kumar; Palanisamy, Rajesh; Harikrishnan, Ramasamy; Arasu, Mariadhas Valan; Al-Dhabi, Naif Abdullah; Arockiaraj, Jesu

    2015-08-01

    In this study, we have reported a molecular characterization of the first B cell lymphoma-2 (BCL-2) related ovarian killer protein (BOK) from freshwater prawn Macrobrachium rosenbergii (Mr). BOK is a novel pro-apoptotic protein of the BCL-2 family that entails in mediating apoptosis to remove cancer cells. A cDNA sequence of MrBOK was identified from the prawn cDNA library and its full length was obtained by internal sequencing. The coding region of MrBOK yields a polypeptide of 291 amino acids. The analysis revealed that MrBOK contains a transmembrane helix at V(261)-L(283) and a putative BCL-2 family domain at V(144)-W(245). MrBOK also possessed four putative BCL-2 homology domains including BH1, BH2, BH3 and weak BH4. The BH3 contains 21 binding sites and among them five residues are highly conserved with the aligned BOK proteins. The homology analysis showed that MrBOK shared maximum similarity with the Caligus rogercresseyi BOK A. The topology of the phylogenetic tree was classified into nine sister groups which includes BOK, BAK, BAX, BAD, BCL-2, BCL-XL, NR13 and MCL members. The BOK protein group further sub-grouped into vertebrate and invertebrate BOK, wherein MrBOK located within insect monophyletic clad of invertebrate BOK. The secondary structural analysis showed that MrBOK contains 11 α-helices (52.2%) which are connected over random coils (47.7%). The 3D structure of MrBOK showed three central helices (α6, α7 and α8) which formed the core of the protein and are flanked on one side by α1, α2 and α3, and on the other side by α4, α5 and α11. MrBOK mRNA is expressed most abundantly (P < 0.05) in ovary compared to other tissues taken for analysis. Hence ovary was selected to study the possible roles of MrBOK mRNA regulation upon bacterial (Aeromonas hydrophila and Vibrio harveyi) and viral [white spot syndrome virus (WSSV) and M. rosenbergii nodovirus] infection. During bacterial and viral infection, the highest MrBOK mRNA transcription was varied at different time points. In bacterial infected ovary tissue, the highest mRNA expression was at 24 h post-infection, whereas in viral infection, the expression was highest at 48 h post-infection. Thus we can conclude that MrBOK functions as an apoptotic protein in intracellular programmed cell-death pathway to counteract the anti-apoptotic proteins released by bacterial and viral pathogens at the time of infection. This is the first study that emphasizes the importance of BOK during bacterial and viral infection in crustacean. PMID:25982403

  11. Development of SYBR Green and TaqMan quantitative real-time PCR assays for hepatopancreatic parvovirus (HPV) infecting Penaeus monodon in India.

    PubMed

    Yadav, Reena; Paria, Anutosh; Mankame, Smruti; Makesh, M; Chaudhari, Aparna; Rajendran, K V

    2015-12-01

    Hepatopancreatic parvovirus (HPV) infects Penaeus monodon and causes mortality in the larval stages. Further, it has been implicated in the growth retardation in cultured P. monodon. Though different geographical isolates of HPV show large sequence variations, a sensitive PCR assay specific to Indian isolate has not yet been reported. Here, we developed a sensitive SYBR Green-based and TaqMan real-time PCR for the detection and quantification of the virus. A 441-bp PCR amplicon was cloned in pTZ57 R/T vector and the plasmid copy number was estimated. A 10-fold serial dilution of the plasmid DNA from 1 × 10(9) copies to 1 copy was prepared and used as the standard. The primers were tested initially using the standard on a conventional PCR format to determine the linearity of detection. The standards were further tested on real-time PCR format using SYBR Green and TaqMan chemistry and standard curves were generated based on the Ct values from three well replicates for each dilution. The assays were found to be sensitive, specific and reproducible with a wide dynamic range (1 × 10(9) to 10 copies) with coefficient of regression (R(2)) > 0.99, calculated average slope -3.196 for SYBR Green assay whereas, for TaqMan assay it was >0.99 and -3.367, respectively. The intra- and inter-assay variance of the Ct values ranged from 0.26% to 0.94% and 0.12% to 0.81%, respectively, for SYBR Green assay, and the inter-assay variance of the Ct values for TaqMan assay ranged from 0.07% to 1.93%. The specificity of the assays was proved by testing other DNA viruses of shrimp such as WSSV, IHHNV and MBV. Standardized assays were further tested to detect and quantify HPV in the post-larvae of P. monodon. The result was further compared with conventional PCR to test the reproducibility of the test. The assay was also used to screen Litopeneaus vannamei, Macrobrachium rosenbergii and Scylla serrata for HPV. PMID:26188128

  12. Orsay virus utilizes ribosomal frameshifting to express a novel protein that is incorporated into virions?

    PubMed Central

    Jiang, Hongbing; Franz, Carl J.; Wu, Guang; Renshaw, Hilary; Zhao, Guoyan; Firth, Andrew E.; Wang, David

    2014-01-01

    Orsay virus is the first identified virus that is capable of naturally infecting Caenorhabditis elegans. Although it is most closely related to nodaviruses, Orsay virus differs from nodaviruses in its genome organization. In particular, the Orsay virus RNA2 segment encodes a putative novel protein of unknown function, termed delta, which is absent from all known nodaviruses. Here we present evidence that Orsay virus utilizes a ribosomal frameshifting strategy to express a novel fusion protein from the viral capsid (alpha) and delta ORFs. Moreover, the fusion protein was detected in purified virus fractions, demonstrating that it is most likely incorporated into Orsay virions. Furthermore, N-terminal sequencing of both the fusion protein and the capsid protein demonstrated that these proteins must be translated from a non-canonical initiation site. While the function of the alphadelta fusion remains cryptic, these studies provide novel insights into the fundamental properties of this new clade of viruses. PMID:24503084

  13. Nanoparticle-based lateral flow biosensor for visual detection of fish nervous necrosis virus amplification products.

    PubMed

    Toubanaki, Dimitra K; Margaroni, Maritsa; Karagouni, Evdokia

    2015-06-01

    Lateral flow paper biosensors are an attractive analytical platform for detection of human and veterinary disease pathogens because they are optimal for accurate, rapid and sensitive analysis in research laboratory setups, as well as field analysis. Since diseases of viral etiology have been wreaking havoc in aquaculture industry, as well as the environment, the present study aims at the development of a gold nanoparticle-based biosensor for fish nervous necrosis virus (Nodavirus) nucleic acids detection. Total viral RNA, isolated from fish samples was subjected to reverse transcription PCR amplification. The PCR products were mixed with a specific oligonucleotide probe and applied next to oligonucleotide conjugated Au NPs. A red test line was formed when nodavirus product was present. The visual detection of the RT-PCR product was completed within 20 min. Following optimization, the biosensor was able to visually detect 270 pg of nodavirus initial total RNA. The present study describes a simple, accurate, robust and low cost method for nodavirus detection in biological samples. Apart contribution on basic research, the proposed biosensor offers great potential for commercial kit development for use on the site of fish culture by fish farmers. This fact will have great impact on environmental safety and disease monitoring without time consuming and costly procedures. PMID:25797786

  14. 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...

  15. Orsay virus utilizes ribosomal frameshifting to express a novel protein that is incorporated into virions

    SciTech Connect

    Jiang, Hongbing; Franz, Carl J.; Wu, Guang; Renshaw, Hilary; Zhao, Guoyan; Firth, Andrew E.; Wang, David

    2014-02-15

    Orsay virus is the first identified virus that is capable of naturally infecting Caenorhabditis elegans. Although it is most closely related to nodaviruses, Orsay virus differs from nodaviruses in its genome organization. In particular, the Orsay virus RNA2 segment encodes a putative novel protein of unknown function, termed delta, which is absent from all known nodaviruses. Here we present evidence that Orsay virus utilizes a ribosomal frameshifting strategy to express a novel fusion protein from the viral capsid (alpha) and delta ORFs. Moreover, the fusion protein was detected in purified virus fractions, demonstrating that it is most likely incorporated into Orsay virions. Furthermore, N-terminal sequencing of both the fusion protein and the capsid protein demonstrated that these proteins must be translated from a non-canonical initiation site. While the function of the alpha–delta fusion remains cryptic, these studies provide novel insights into the fundamental properties of this new clade of viruses. - Highlights: • Orsay virus encodes a novel fusion protein by a ribosomal frameshifting mechanism. • Orsay capsid and fusion protein is translated from a non-canonical initiation site. • The fusion protein is likely incorporated into Orsay virions.

  16. Regulation of natural killer enhancing factor (NKEF) genes in teleost fish, gilthead seabream and European sea bass.

    PubMed

    Esteban, Mara A; Chaves-Pozo, Elena; Arizcun, Marta; Meseguer, Jos; Cuesta, Alberto

    2013-10-01

    Peroxiredoxins (Prx) are a family of antioxidant proteins also involved in inflammation and innate immunity. Prx1 and Prx2 are also known as natural killer enhancing factor (NKEF)-A and NKEF-B, respectively, by their ability to prime the mammalian NK-cells activity. In teleost fish, NKEF genes have been isolated but their regulation has been scarcely evaluated. We have identified orthologues of the NKEF-A and NKEF-B genes in the teleost European sea bass (Dicentrarchus labrax) which showed constitutive expression and wide distribution in their tissues. In vitro, the gilthead seabream (Sparus aurata) and sea bass NKEFs were slightly up-regulated in head-kidney leucocytes after stimulation with unmethylated CpG oligodeoxynucleotides, poly I:C or pathogenic bacteria. In vivo, seabream and sea bass infection with nodavirus up-regulated the expression of NKEF genes in the brain (target tissue for nodavirus) and head-kidney at different infection times. Although further studies are necessary to ascertain their role as antioxidant proteins and in the immune response in teleost fish, our results suggest a primary role of seabream and sea bass NKEFs in the innate immune response against bacterial and viral agents. PMID:23511025

  17. Optimisation of Lime-Soda process parameters for reduction of hardness in aqua-hatchery practices using Taguchi methods.

    PubMed

    Yavalkar, S P; Bhole, A G; Babu, P V Vijay; Prakash, Chandra

    2012-04-01

    This paper presents the optimisation of Lime-Soda process parameters for the reduction of hardness in aqua-hatchery practices in the context of M. rosenbergii. The fresh water in the development of fisheries needs to be of suitable quality. Lack of desirable quality in available fresh water is generally the confronting restraint. On the Indian subcontinent, groundwater is the only source of raw water, having varying degree of hardness and thus is unsuitable for the fresh water prawn hatchery practices (M. rosenbergii). In order to make use of hard water in the context of aqua-hatchery, Lime-Soda process has been recommended. The efficacy of the various process parameters like lime, soda ash and detention time, on the reduction of hardness needs to be examined. This paper proposes to determine the parameter settings for the CIFE well water, which is pretty hard by using Taguchi experimental design method. Orthogonal Arrays of Taguchi, Signal-to-Noise Ratio, the analysis of variance (ANOVA) have been applied to determine their dosage and analysed for their effect on hardness reduction. The tests carried out with optimal levels of Lime-Soda process parameters confirmed the efficacy of the Taguchi optimisation method. Emphasis has been placed on optimisation of chemical doses required to reduce the total hardness using Taguchi method and ANOVA, to suit the available raw water quality for aqua-hatchery practices, especially for fresh water prawn M. rosenbergii. PMID:24749379

  18. Nonlytic spread of naked viruses.

    PubMed

    Bird, Sara W; Kirkegaard, Karla

    2015-01-01

    How do viruses spread from cell to cell? Enveloped viruses acquire their surrounding membranes by budding: either through the plasma membrane or an internal membrane of infected cells. Thus, a newly budded enveloped virus finds itself either in the extracellular milieu or in a lumenal compartment from which it can exit the cell by conventional secretion. On the other hand, naked viruses such as poliovirus, nodavirus, adenovirus, and SV40 lack an external membrane. They are simply protein-nucleic acid complexes within the cytoplasm or nucleus of the infected cell, and thus would seem to have no other exit route than cell lysis. We have presented the first documentation of nonlytic spread of a naked virus, and showed the interconnections between this event and the process or components of the autophagy pathway. PMID:25680079

  19. Outbreak of betanodavirus infection in tilapia, Oreochromis niloticus (L.), in fresh water.

    PubMed

    Bigarré, L; Cabon, J; Baud, M; Heimann, M; Body, A; Lieffrig, F; Castric, J

    2009-08-01

    A betanodavirus associated with a massive mortality was isolated from larvae of tilapia, Oreochromis niloticus, maintained in fresh water at 30 degrees C. Histopathology revealed vacuolation of the nervous system, suggesting an infection by a betanodavirus. The virus was identified by indirect fluorescent antibody test in the SSN1 cell line and further characterized by sequencing of a PCR product. Sequencing of the T4 region of the coat protein gene indicated a phylogenetic clustering of this isolate within the red-spotted grouper nervous necrosis virus type. However, the tilapia isolate formed a unique branch distinct from other betanodavirus isolates. The disease was experimentally reproduced by bath infection of young tilapia at 30 degrees C. The reservoir of virus at the origin of the outbreak remains unidentified. To our knowledge, this is the first report of natural nodavirus infection in tilapia reared in fresh water. PMID:19500206

  20. What They Work at is not What They eat: Incongruence of Strong Interactions and Assimilation in Shrimps and Baetid Mayflies in a Brazilian Coastal Stream

    NASA Astrophysics Data System (ADS)

    Moulton, T.; de Souza, M. L.; Brito, E. F.; Krsulovic, F. M.; Silveira, R. M.; Ometto, J. B.

    2005-05-01

    Exclusion experiments showed that atyid shrimps (Potimirim glabra) and baetid mayflies (Cloeodes sp., Americabaetis sp.) removed benthic material from hard substrate at different sites in the forested stream Rio Andorinha, Ilha Grande, Rio de Janeiro. Their effects were greater on organic material not associated with algae than on algae. Macrobrachium olfersi shrimp had a negative effect on the mayflies, causing a trophic cascade with the benthic material. Gut analysis of the atyid and baetids showed that they ingested large amounts of detritus, as expected. Macrobrachium contained mainly detritus, some algae and some insect parts. Stable isotope analysis of the gut contents confirmed these compositions, but the tissue of the shrimps and mayflies was of carbon derived from algae (as testified by enriched δ 13C corresponding to microalgae from the periphyton). Baetids appeared to be algal herbivores and both Potimirim and Macrobrachium appeared (from their δ 15N signatures) to be secondary consumers. We note that what the animals remove (both by bioturbation and ingestion) and what they assimilate are very different. We speculate that the baetids and atyid are "gardeners" and remove material to promote the growth of algal turf.

  1. 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

  2. Use of intertidal areas by shrimps (Decapoda) in a Brazilian Amazon estuary.

    PubMed

    Sampaio, Hebert A; Martinelli-Lemos, Jussara M

    2014-03-01

    The present work investigated the occupation and the correlation of the shrimp abundance in relation to environmental variables in different habitats (mangroves, salt marshes and rocky outcrops) in an Amazon estuary. The collections were made in August and November 2009, at low syzygy tide on Areuá Beach, situated in the Extractive Reserve of Mãe Grande de Curuçá, Pará, Brazil totaling 20 pools. In each environment, we recorded the physical-chemical factors (pH, salinity, and temperature) and measured the area (m2) and volume (m3) of every pool through bathymetry. The average pH, salinity, temperature, area and volume of tide pools were 8.75 (± 0.8 standard deviation), 35.45 (± 3), 29.49 °C (± 2.32), 27.41 m2 (± 41.18), and 5.19 m3 (± 8.01), respectively. We caught a total of 4,871 shrimps, distributed in three families and four species: Farfantepenaeus subtilis (98.36%) (marine) followed by Alpheus pontederiae (0.76%) (estuarine), Macrobrachium surinamicum (0.45%) and Macrobrachium amazonicum (0.43%) predominantly freshwater. The species F. subtilis and A. pontederiae occurred in the three habitats, whereas M. surinamicum occurred in salt marsh and rocky outcrop and M. amazonicum only in marisma. Temperature and pH were the most important environmental descriptors that significantly affected the density and biomass of shrimps. PMID:24676171

  3. 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

  4. 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

  5. Genomic RNA of an insect virus directs synthesis of infectious virions in plants.

    PubMed Central

    Selling, B H; Allison, R F; Kaesberg, P

    1990-01-01

    Newly synthesized virions of flock house virus (FHV), an insect nodavirus, were detected in plant cells inoculated with FHV RNA. FHV was found in whole plants of barley (Hordeum vulgare), cowpea (Vigna sinensis), chenopodium (Chenopodium hybridum), tobacco (Nicotiana tabacum), and Nicotiana benthamiana and in protoplasts derived from barley leaves. Virions produced in plants contained newly synthesized RNA as well as newly synthesized capsid protein. These results show that the intracellular environment in these plants is suitable for synthesis of a virus normally indigenous only to insects. Such synthesis involves, minimally, translation of viral RNA, RNA replication, and virion assembly. Inoculation of barley protoplasts with FHV virions resulted in synthesis of small amounts of progeny virions, suggesting that FHV virions are capable of releasing their RNA in plant cells. In N. benthamiana, virions resulting from inoculation with RNA were detected not only in inoculated leaves but also in other leaves of inoculated plants, suggesting that virions could move in this plant species. Such movement probably occurs by a passive transport through the vascular system rather than by an active transport involving mechanisms that have evolved for plant viruses. Images PMID:2296598

  6. Biomonitoring of Cd, Cr, Hg and Pb in the Baluarte River basin associated to a mining area (NW Mexico).

    PubMed

    Ruelas-Inzunza, J; Green-Ruiz, C; Zavala-Nevárez, M; Soto-Jiménez, M

    2011-08-15

    With the purpose of knowing seasonal variations of Cd, Cr, Hg and Pb in a river basin with past and present mining activities, elemental concentrations were measured in six fish species and four crustacean species in Baluarte River, from some of the mining sites to the mouth of the river in the Pacific Ocean between May 2005 and March 2006. In fish, highest levels of Cd (0.06 μg g ⁻¹ dry weight) and Cr (0.01 μg g⁻¹) were detected during the dry season in Gobiesox fluviatilis and Agonostomus monticola, respectively; the highest levels of Hg (0.56 μg g⁻¹) were detected during the dry season in Guavina guavina and Mugil curema. In relation to Pb, the highest level (1.65 μg g⁻¹) was detected in A. monticola during the dry season. In crustaceans, highest levels of Cd (0.05 μg g⁻¹) occurred in Macrobrachium occidentale during both seasons; highest concentration of Cr (0.09 μg g⁻¹) was also detected in M. occidentale during the dry season. With respect to Hg, highest level (0.20 μg g⁻¹) was detected during the rainy season in Macrobrachium americanum; for Pb, the highest concentration (2.4 μg g⁻¹) corresponded to Macrobrachium digueti collected in the dry season. Considering average concentrations of trace metals in surficial sediments from all sites, Cd (p<0.025), Cr (p<0.10) and Hg (p<0.15) were significantly higher during the rainy season. Biota sediment accumulation factors above unity were detected mostly in the case of Hg in fish during both seasons. On the basis of the metal levels in fish and crustacean and the provisional tolerable weekly intake of studied elements, people can eat up to 13.99, 0.79 and 2.34 kg of fish in relation to Cd, Hg and Pb, respectively; regarding crustaceans, maximum amounts were 11.33, 2.49 and 2.68 kg of prawns relative to levels of Cd, Hg and Pb, respectively. PMID:21684575

  7. [Ecophysiological adaptability of tropical water organisms to salinity changes].

    PubMed

    Chung, K S

    2001-03-01

    Physiological response of tropical organisms to salinity changes was studied for some marine, estuarine and freshwater fishes (Astyanax bimaculatus, Petenia karussii, Cyprinodon dearborni, and Oreochromis mossambicus), marine and freshwater crustaceans (Penaeus brasiliensis, Penaeus schmitti and Macrobrachium carcinus), and marine bivalves (Perna perna, Crassostrea rhizophorae, and Arca zebra) collected from Northeast Venezuela. They were acclimated for four weeks at various salinities, and (1) placed at high salinities to determine mean lethal salinity, (2) tested by increasing salinity 5@1000 per day to define upper lethal salinity tolerance limit, or (3) observed in a saline gradient tank to determine salinity preference. Acclimation level was the most significant factor. This phenomenon is important for tropical aquatic organisms in shallow waters, where they can adapt to high salinity during the dry season and cannot lose their acclimation level at low salinity during abrupt rain. For saline adaptation of tropical organisms, this behavior will contribute to their proliferation and distribution in fluctuating salinity environments. PMID:11795174

  8. Water temperature affects pathogenicity of different betanodavirus genotypes in experimentally challenged Dicentrarchus labrax.

    PubMed

    Toffan, Anna; Panzarin, Valentina; Toson, Marica; Cecchettin, Krizia; Pascoli, Francesco

    2016-05-26

    Betanodaviruses are the causative agents of a highly infectious disease of fish known as viral nervous necrosis (VNN). To date, 4 different nervous necrosis virus (NNV) genotypes have been described, but natural reassortant viruses have also been detected, which further increase viral variability. Water temperature plays an important role in determining the appearance and the severity of VNN disease. We assessed the effect of temperature (20°, 25° and 30°C) on mortality and virus load in the brain of European sea bass Dicentrarchus labrax experimentally infected with 4 genetically different betanodaviruses, namely red-spotted grouper NNV (RGNNV), striped jack NNV (SJNNV) and the reassortant strains RGNNV/SJNNV and SJNNV/RGNNV. The RGNNV/SJNNV virus possesses the polymerase gene of RGNNV and the coat protein gene of SJNNV, and vice versa for the SJNNV/RGNNV virus. The obtained results showed that the RGNNV strain is the most pathogenic for juvenile sea bass, but clinical disease and mortality appeared only at higher temperatures. The SJNNV strain is weakly pathogenic for D. labrax regardless of the temperature used, while virus replication was detected in the brain of survivors only at 20°C. Finally, reassortant strains caused low mortality, independent of the temperature used, but the viral load in the brain was strongly influenced by water temperature and the genetic type of the polymerase gene. Taken together, these data show that nodavirus replication in vivo is a composite process regulated by both the genetic features of the viral strain and water temperatures. PMID:27225206

  9. Infectious bursal disease virus capsid assembly and maturation by structural rearrangements of a transient molecular switch.

    PubMed

    Luque, Daniel; Saugar, Irene; Rodríguez, José F; Verdaguer, Nuria; Garriga, Damiá; Martín, Carmen San; Velázquez-Muriel, Javier A; Trus, Benes L; Carrascosa, José L; Castón, José R

    2007-07-01

    Infectious bursal disease virus (IBDV), a double-stranded RNA (dsRNA) virus belonging to the Birnaviridae family, is an economically important avian pathogen. The IBDV capsid is based on a single-shelled T=13 lattice, and the only structural subunits are VP2 trimers. During capsid assembly, VP2 is synthesized as a protein precursor, called pVP2, whose 71-residue C-terminal end is proteolytically processed. The conformational flexibility of pVP2 is due to an amphipathic alpha-helix located at its C-terminal end. VP3, the other IBDV major structural protein that accomplishes numerous roles during the viral cycle, acts as a scaffolding protein required for assembly control. Here we address the molecular mechanism that defines the multimeric state of the capsid protein as hexamers or pentamers. We used a combination of three-dimensional cryo-electron microscopy maps at or close to subnanometer resolution with atomic models. Our studies suggest that the key polypeptide element, the C-terminal amphipathic alpha-helix, which acts as a transient conformational switch, is bound to the flexible VP2 C-terminal end. In addition, capsid protein oligomerization is also controlled by the progressive trimming of its C-terminal domain. The coordination of these molecular events correlates viral capsid assembly with different conformations of the amphipathic alpha-helix in the precursor capsid, as a five-alpha-helix bundle at the pentamers or an open star-like conformation at the hexamers. These results, reminiscent of the assembly pathway of positive single-stranded RNA viruses, such as nodavirus and tetravirus, add new insights into the evolutionary relationships of dsRNA viruses. PMID:17442720

  10. Dual roles for an arginine-rich motif in specific genome recognition and localization of viral coat protein to RNA replication sites in flock house virus-infected cells.

    PubMed

    Venter, P Arno; Marshall, Dawn; Schneemann, Anette

    2009-04-01

    Assembly of many RNA viruses entails the encapsidation of multiple genome segments into a single virion, and underlying mechanisms for this process are still poorly understood. In the case of the nodavirus Flock House virus (FHV), a bipartite positive-strand RNA genome consisting of RNA1 and RNA2 is copackaged into progeny virions. In this study, we investigated whether the specific packaging of FHV RNA is dependent on an arginine-rich motif (ARM) located in the N terminus of the coat protein. Our results demonstrate that the replacement of all arginine residues within this motif with alanines rendered the resultant coat protein unable to package RNA1, suggesting that the ARM represents an important determinant for the encapsidation of this genome segment. In contrast, replacement of all arginines with lysines had no effect on RNA1 packaging. Interestingly, confocal microscopic analysis demonstrated that the RNA1 packaging-deficient mutant did not localize to mitochondrial sites of FHV RNA replication as efficiently as wild-type coat protein. In addition, gain-of-function analyses showed that the ARM by itself was sufficient to target green fluorescent protein to RNA replication sites. These data suggest that the packaging of RNA1 is dependent on trafficking of coat protein to mitochondria, the presumed site of FHV assembly, and that this trafficking requires a high density of positive charge in the N terminus. Our results are compatible with a model in which recognition of RNA1 and RNA2 for encapsidation occurs sequentially and in distinct cellular microenvironments. PMID:19158251

  11. 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.

  12. 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

  13. 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

  14. Establishment and characterization of fin cell lines from diploid, triploid, and tetraploid oriental weatherfish (Misgurnus anguillicaudatus).

    PubMed

    Li, Xia; Ma, Chen; Qin, Yan-Jie; Li, Ya-Juan; Wu, Di; Bai, Li-Wen; Pei, Ai-Jun

    2015-06-01

    Continuous fin cell lines from diploid, triploid, and tetraploid oriental weatherfish, Misgurnus anguillicaudatus, were established and characterized. The cell lines, designated DIMF, TRMF, and TEMF, respectively, were subcultured more than 80 times since initiation in October 2012 and were preserved at the China Center for Type Culture Collection as sample numbers C2013109, C2013110, C2013111, respectively. The cell lines consist predominantly of fibroblast-like cells. At the 50th passage, the population doubling times were 48.43 h (DIMF), 36.01 h (TRMF), and 41.45 h (TEMF). Cell survival rate of these three kinds of cells was 80.88 ± 1.38, 84.48 ± 1.13, and 81.57 ± 1.28 %, respectively, when recovered after storage in liquid nitrogen for 60 days at the 40th passage. The chromosome numbers measured from 100 metaphase plates at the 50th passage were 2n = 50 (68 %), 3n = 75 (59 %), and 4n = 100 (54 %) for DIMF, TRMF, and TEMF cells, respectively. At the 60th passage, the chromosome numbers for DIMF and TRMF cells were still 50 (52 %) and 75 (70 %), but the chromosome number for TEMF cells ranged from 88 to 100; a chromosome number of 96 accounted for 26 % of the cells, and the karyotype analysis showed 4n = 96, 16 m + 8sm + 72t, NF = 120; thus, compared with cells at the 50th passage, a group of metacentric chromosomes was missing. Microsatellite marker analysis was conducted using DIMF, TRMF, and TEMF cells and muscle tissue of oriental weatherfish, which confirmed that the three cell lines established in this study were from oriental weatherfish. The cell lines were exposed to two fish viruses to determine their susceptibility to infection; they were susceptible to spring viremia of carp virus but not to piscine nodavirus. Establishment of fin cell lines from different ploidy oriental weatherfish increases the existing number of fish cell lines available for research, and it provides a model for investigating the mechanisms of growth and genetics in polyploid fish. PMID:25721100

  15. Identification of six novel CC chemokines in gilthead seabream (Sparus aurata) implicated in the antiviral immune response.

    PubMed

    Cuesta, A; Dios, S; Figueras, A; Novoa, B; Esteban, M A; Meseguer, J; Tafalla, C

    2010-03-01

    Chemokines are key regulators of migration and consequent activation of migrating leukocytes. CC chemokines constitute the largest chemokine group with 24-28 members in mammalian species, and even more in teleost fish, with up to 81 members in zebrafish Danio rerio. Further studies concerning fish chemokine genes will help elucidate the complexity of this chemokine group which has considerably expanded in some teleosts. In the current work, we have identified six novel CC chemokine genes within previously generated gilthead seabream (Sparus aurata) EST databases. The six novel chemokine sequences all show characteristic features of CC chemokines, such as the 4 conserved cysteine residues and a signal peptide. The nomenclature for chemokines in different fish species is not in concordance to mammalian nomenclature as it is difficult to establish true mammalian orthologs, and therefore a different nomenclature has been established for each fish species. In this work, we have named the seabream genes according to the rainbow trout CC chemokine with which they have the highest identity, therefore we have designated the novel seabream CC chemokines as CK1, CK3, CK5, CK7, CK8 and CK10. Expression analysis have also been performed with these new chemokines, as well as with the previously identified seabream chemokine designated as CCL4, which according to our proposed nomenclature should be renamed CK5B. In this sense, we have determined the pattern of constitutive chemokine expression in different seabream tissues. The effect that different immune non-replicative stimuli had in the levels of expression of the chemokines in head kidney leukocytes showed many strong suppressive effects in their transcription levels, and up-regulations mainly in response to mitogens. In vivo, however, when non-replicative virus or heat-killed bacteria were injected, the viral particles up-regulated chemokine expression in the spleen and not in head kidney. Finally, in the context of a real infection such as that of nodavirus in the brain, all the CC chemokines studied were significantly induced. This study constitutes a further step towards the elucidation of an immunological and/or physiological role for fish chemokines. PMID:20096460

  16. Infectious Bronchitis Virus Generates Spherules from Zippered Endoplasmic Reticulum Membranes

    PubMed Central

    Maier, Helena J.; Hawes, Philippa C.; Cottam, Eleanor M.; Mantell, Judith; Verkade, Paul; Monaghan, Paul; Wileman, Tom; Britton, Paul

    2013-01-01

    ABSTRACT Replication of positive-sense RNA viruses is associated with the rearrangement of cellular membranes. Previous work on the infection of tissue culture cell lines with the betacoronaviruses mouse hepatitis virus and severe acute respiratory syndrome coronavirus (SARS-CoV) showed that they generate double-membrane vesicles (DMVs) and convoluted membranes as part of a reticular membrane network. Here we describe a detailed study of the membrane rearrangements induced by the avian gammacoronavirus infectious bronchitis virus (IBV) in a mammalian cell line but also in primary avian cells and in epithelial cells of ex vivo tracheal organ cultures. In all cell types, structures novel to IBV infection were identified that we have termed zippered endoplasmic reticulum (ER) and spherules. Zippered ER lacked luminal space, suggesting zippering of ER cisternae, while spherules appeared as uniform invaginations of zippered ER. Electron tomography showed that IBV-induced spherules are tethered to the zippered ER and that there is a channel connecting the interior of the spherule with the cytoplasm, a feature thought to be necessary for sites of RNA synthesis but not seen previously for membrane rearrangements induced by coronaviruses. We also identified DMVs in IBV-infected cells that were observed as single individual DMVs or were connected to the ER via their outer membrane but not to the zippered ER. Interestingly, IBV-induced spherules strongly resemble confirmed sites of RNA synthesis for alphaviruses, nodaviruses, and bromoviruses, which may indicate similar strategies of IBV and these diverse viruses for the assembly of RNA replication complexes. IMPORTANCE All positive-sense single-stranded RNA viruses induce rearranged cellular membranes, providing a platform for viral replication complex assembly and protecting viral RNA from cellular defenses. We have studied the membrane rearrangements induced by an important poultry pathogen, the gammacoronavirus infectious bronchitis virus (IBV). Previous work studying closely related betacoronaviruses identified double-membrane vesicles (DMVs) and convoluted membranes (CMs) derived from the endoplasmic reticulum (ER) in infected cells. However, the role of DMVs and CMs in viral RNA synthesis remains unclear because these sealed vesicles lack a means of delivering viral RNA to the cytoplasm. Here, we characterized structures novel to IBV infection: zippered ER and small vesicles tethered to the zippered ER termed spherules. Significantly, spherules contain a channel connecting their interior to the cytoplasm and strongly resemble confirmed sites of RNA synthesis for other positive-sense RNA viruses, making them ideal candidates for the site of IBV RNA synthesis. PMID:24149513

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

    PubMed Central

    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.

    2015-01-01

    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. PMID:26195752

  18. DDT and Derivatives in Indicator Species of the Aquatic Food Web of Rangsit Agricultural Area, Central Thailand

    PubMed Central

    Siriwong, W.; Thirakhupt, K.; Sitticharoenchai, D.; Rohitrattana, J.; Thongkongowm, P.; Borjan, M.; Robson, M.

    2009-01-01

    The presence of DDT and derivatives in the food web of freshwater ecosystems of Rangsit agricultural area, Pathum Thani Province, Thailand were investigated from June 2004 to May 2007. By using gas chromatography (GC) with micro electron capture detector (μ ECD), DDT and derivatives in water, sediment, and fifteen indicator species i.e., 2 producers; Eichhornia crassipes and plankton (phyto- and zoo- plankton), an herbivore; Trichogaster microlepis (3) 3 omnivores; Trichogaster trichopterus, Oreochromis niloticus, and Puntius gonionotus, 6 carnivores; Channa striatus, Oxyeleotris marmoratus, Macrognathus siamensis, Parambassis siamensis, Anabas testudineus, and Pristolepis fasciatus, and 3 detritivores; Macrobrachium lanchesteri, Pomacea sp., and Filopaludina mertensi were measured. Results show low concentration levels (part per billion) of DDT & derivatives in each food web compartment i.e. water, sediment, aquatic plant, plankton, fish, and invertebrates. Magnification patterns, i.e. bioconcentration, bioaccumulation, and biomagnification, based on habitat and foraging behavior of selected freshwater species indicates that DDT & derivatives can accumulate and be magnified through the food chain from the lowest up to the highest trophic level. Therefore, the presence of residues and the evidence of magnification patterns can be observed as ecological indicators for evaluating ecological health risk. PMID:20161116

  19. Predator-prey imbalances due to a pesticide: density and applicability timing as determining factors for experimental assessments.

    PubMed

    Gutierrez, María Florencia; Negro, Carlos Leandro

    2014-09-01

    Predator-prey relationships are determining factors in sustaining community structure but xenobiotics, including pesticides, have the potential to alter them, causing imbalances at the ecosystem level. Although invertebrate predation on zooplankton is of high importance in shallow lakes, there is still little information regarding disturbances on this trophic interaction. This work assessed the potential effects of a chlorpyrifos-based pesticide (CLP) on the interaction between prawns Macrobrachium borellii and cladocerans Ceriodaphnia dubia, taking into account prey densities, specific time of exposure and contamination level. The analysis was focused on the specific sensitivity of both species and, especially, on the predation rate of M. borellii on C. dubia. The latter was evaluated through different treatments that combined predator and/or prey exposure to the insecticide, before (lapse of 12 h) or during the interaction. Under low prey density, when prawns were previously exposed to the insecticide, their consumption rate was lower than that of controls. Conversely, when cladocerans or both species were previously exposed, the prawns' feeding rate was higher. Under high prey density, there were no substantial differences among treatments. Comparatively, cladocerans were significantly more consumed when the exposure of both species was performed before rather than during the interaction. From the results obtained, it can be assumed that the trophic interaction under study is sensitive to CLP and that individual density and specific time of exposure are important variables to be considered in similar studies in order to obtain realistic results. PMID:24903805

  20. Infection with schistosome parasites in snails leads to increased predation by prawns: implications for human schistosomiasis control.

    PubMed

    Swartz, Scott J; De Leo, Giulio A; Wood, Chelsea L; Sokolow, Susanne H

    2015-12-01

    Schistosomiasis - a parasitic disease that affects over 200 million people across the globe - is primarily transmitted between human definitive hosts and snail intermediate hosts. To reduce schistosomiasis transmission, some have advocated disrupting the schistosome life cycle through biological control of snails, achieved by boosting the abundance of snails' natural predators. But little is known about the effect of parasitic infection on predator-prey interactions, especially in the case of schistosomiasis. Here, we present the results of laboratory experiments performed on Bulinus truncatus and Biomphalaria glabrata snails to investigate: (i) rates of predation on schistosome-infected versus uninfected snails by a sympatric native river prawn, Macrobrachium vollenhovenii, and (ii) differences in snail behavior (including movement, refuge-seeking and anti-predator behavior) between infected and uninfected snails. In predation trials, prawns showed a preference for consuming snails infected with schistosome larvae. In behavioral trials, infected snails moved less quickly and less often than uninfected snails, and were less likely to avoid predation by exiting the water or hiding under substrate. Although the mechanism by which the parasite alters snail behavior remains unknown, these results provide insight into the effects of parasitic infection on predator-prey dynamics and suggest that boosting natural rates of predation on snails may be a useful strategy for reducing transmission in schistosomiasis hotspots. PMID:26677260

  1. Environmental features and macrofauna of Kahana Estuary, Oahu, Hawaii

    USGS Publications Warehouse

    Maciolek, J.A.; Timbol, A.S.

    1981-01-01

    Lack of ecological information on Hawaiian estuaries prompted an intensive 2-year study of a small (5.7 ha) stream-mouth estuary on windward Oahu. Water quality and macrofauna were sampled weekly at seven stations. The water mass was strongly stratified vertically except during freshets. Average values for water column temperature and bottom salinity were 23.2°C and 12‰ at the head to 28.3°C and 28‰ at the mouth. Dissolved oxygen saturation in the water column varied from about 50% at night to 140% in the afternoon. Usually, bottom waters were 3–6°C warmer than surface waters and sometimes showed severe oxygen depletion.Macrofauna, collected primarily by seining, consisted mainly of decapod crustaceans (four species of crabs, seven species of shrimps) and fishes (24 species). Other typical estuarine taxons (mollusks, barnacles, polychaetes) were scarce or absent. Diversity increased seaward from 14 species near the estuary head to 29 species near the mouth. Three species of crustaceans and six of fishes were captured at all stations. Most abundant were the native prawn, Macrobrachium grandimanus, and mullet, Mugil cephalus. Perennially resident adults occurred among crustaceans and gobioid fishes; most other fishes were present as juveniles and sporadic adults. Comparisons with other data suggest that more than 50 species of native fishes may occur in Hawaiian estuaries, and that estuarine macrofaunal diversity on oceanic islands is much lower than on continents at similar latitudes.

  2. One-step PCR amplification of complete arthropod mitochondrial genomes.

    PubMed

    Hwang, U W; Park, C J; Yong, T S; Kim, W

    2001-06-01

    A new PCR primer set which enables one-step amplification of complete arthropod mitochondrial genomes was designed from two conserved 16S rDNA regions for the long PCR technique. For this purpose, partial 16S rDNAs amplified with universal primers 16SA and 16SB were newly sequenced from six representative arthropods: Armadillidium vulgare and Macrobrachium nipponense (Crustacea), Anopheles sinensis (Insecta), Lithobius forficatus and Megaphyllum sp. (Myriapoda), and Limulus polyphemus (Chelicerata). The genomic locations of two new primers, HPK16Saa and HPK16Sbb, correspond to positions 13314-13345 and 12951-12984, respectively, in the Drosophila yakuba mitochondrial genome. The usefulness of the primer set was experimentally examined and confirmed with five of the representative arthropods, except for A. vulgare, which has a linearized mitochondrial genome. With this set, therefore, we could easily and rapidly amplify complete mitochondrial genomes with small amounts of arthropod DNA. Although the primers suggested here were examined only with arthropod groups, a possibility of successful application to other invertebrates is very high, since the high degree of sequence conservation is shown on the primer sites in other invertebrates. Thus, this primer set can serve various research fields, such as molecular evolution, population genetics, and molecular phylogenetics based on DNA sequences, RFLP, and gene rearrangement of mitochondrial genomes in arthropods and other invertebrates. PMID:11399145

  3. 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.                            

  4. [Predator damage and shell size on the diadromous snail Neritina virginea (Gastropoda: Neritidae) in the Mameyes River, Puerto Rico].

    PubMed

    Blanco-Libreros, Juan Felipe; Arroyave-Rincón, Andrea

    2009-12-01

    We compared predators' damage with shell size in live individuals and empty shells (n=5066) of the snail Neritina virginea in the Mameyes River (Puerto Rico, Greater Antilles). According to the literature and direct observations, damages on empty shells were attributed to predation by aquatic birds (e.g. Gallinula chloropus) and decapods (e.g. Macrobrachium spp.), while damages on live individuals were due to rasping by co-specifics and erosion. Predation by decapods and birds, as estimated by the proportion of empty shells, was low (2 and 0.36%, respectively). Shell size was significantly different between types of predators (range: decapods: 3.5-15.0mm, birds: 8.1-19.4mm). By comparing sizes of the empty shells and the live individuals, we concluded that decapods specialize on large groups of small migratory juveniles, while birds specialize on the largest resident individuals. Worn shells were highly frequent in both empty shells and live individuals, and sizes did not differ between samples. A comparison by slow-flow and fast-flow habitats showed that predators do not discriminate shell sizes between environments. However, the frequency of damage by birds and decapods was greater under slow-flow conditions. Despite of the little contribution of predation to the population dynamics in this species, predation might be an important driver of size-dependent behavioral responses such as upstream migration and microhabitat selection. PMID:20073335

  5. 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

  6. 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

  7. 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

  8. 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

  9. Paragonimiasis and its intermediate hosts in a transmission focus in Lao People's Democratic Republic.

    PubMed

    Odermatt, P; Habe, S; Manichanh, S; Tran, D S; Duong, V; Zhang, Wei; Phommathet, K; Nakamura, S; Barennes, H; Strobel, M; Dreyfuss, G

    2007-08-01

    Human cases of paragonimiasis have been reported in Lao People's Democratic Republic from several provinces, however, detailed descriptions of a focus of transmission including parasites species and intermediate hosts are lacking. Field investigations were carried out between February and July 2004 in three neighboring villages of the province of Vientiane. Persons with chronic cough (more than three weeks) were identified during a door-to-door survey and three sputum samples were examined for Paragonimus spp. and tuberculosis on two consecutive days. Potential intermediate snail and crustacean hosts were sampled among the populations' water-contact sites and examined for metacercarial infections or cercarial shedding. Four dogs were infected with metacercaria and the species of the subsequently retrieved adult parasites were identified. 12.7% (15 cases) of all chronic cough patients (0.6% of the total population) suffered from paragonimiasis. Samples of five species of freshwater snails, two species of crabs and one species of prawn were collected from the Nam Sêt River. None of the snails were infected. Metacercariae of P. harinasutai, P. bangkokensis and P. heterotremus were found in Potamon lipkei crabs while an infection with P. westermani was observed in a Chulathelphusa brandti crab. No paragonimid metacercaria was found in the Macrobrachium dienbienphuense prawns Dang and Nguyen 1972. This study demonstrates the importance of Paragonimus spp. as an etiological agent in patients with chronic cough and suggests that similar transmission might be widespread in Lao People's Democratic Republic. We document herein the transmission of several Paragonimus species indicating that paragonimiasis is an active zoonosis in Lao People's Democratic Republic. PMID:17626777

  10. 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

  11. Cloning of genomic sequences of three crustacean hyperglycemic hormone superfamily genes and elucidation of their roles of regulating insulin-like androgenic gland hormone gene.

    PubMed

    Li, Fajun; Bai, Hongkun; Zhang, Wenyi; Fu, Hongtuo; Jiang, Fengwei; Liang, Guoxia; Jin, Shubo; Sun, Shengming; Qiao, Hui

    2015-04-25

    The insulin-like androgenic gland hormone (IAG) gene in crustaceans plays an important role in male sexual differentiation, metabolism, and growth. However, the upstream regulation of IAG signaling schemes remains poorly studied. In the present study, we cloned the 5' flanking sequence of IAG and full-length genomic sequences of gonad-inhibiting hormone (Mn-GIH), molt-inhibiting hormone (Mn-MIH) and crustacean hyperglycemic hormone (Mn-CHH) in Macrobrachium nipponense. We identified the transcription factor-binding sites in the 5' flanking sequence of IAG and investigated the exon-intron patterns of the three CHH superfamily genes. Each CHH superfamily gene consisted of two introns separating three exons. Mn-GIH and Mn-MIH shared the same intron insertion sites, which differed from Mn-CHH. We provided DNA-level evidence for the type definition. We also identified two cAMP response elements in the 5' untranslated region. We further investigated the regulatory relationships between Mn-GIH, Mn-MIH, and Mn-CHH and IAG at the transcriptional level by injection of double-stranded RNA (dsRNA). IAG transcription levels were significantly increased to 660.2%, 472.9%, and 112.4% of control levels in the Mn-GIH dsRNA, Mn-MIH dsRNA, and Mn-CHH dsRNA groups, respectively. The results clearly demonstrated that Mn-GIH and Mn-MIH, but not Mn-CHH, negatively regulate the expression of the IAG gene. PMID:25680292

  12. Do osmoregulators have lower capacity of muscle water regulation than osmoconformers? A study on decapod crustaceans.

    PubMed

    Foster, Clarice; Amado, Enelise M; Souza, Marta M; Freire, Carolina A

    2010-02-01

    Decapod crustaceans occupy various aquatic habitats. In freshwater they are osmoregulators, while marine species are typically osmoconformers. Freshwater crustaceans are derived from marine ancestors. The hypothesis tested here was that osmoregulators, which can rely on salt transport by interface epithelia to prevent extracellular disturbance, would have a lower capacity of tissue water regulation when compared with osmoconformers. Four species of decapod crustaceans (the marine osmoconformer crab Hepatus pudibundus, and three osmoregulators of different habitats) have been exposed in vivo to a salinity challenge, for up to 24 hr. Osmoregulators were: the estuarine shrimp Palaemon pandaliformis, the diadromous freshwater shrimp Macrobrachium acanthurus, and the hololimnetic red crab Dilocarcinus pagei. H. pudibundus displayed hemolymph dilution already after 0.5 hr in 25 per thousand, reaching approximately 30% reduction in osmolality, but its muscle degree of hydration did not increase. To make the different in vivo salinity challenges directly comparable, the ratio between the maximum change in muscle hydration with respect to the control value measured for the species and the maximum change in hemolymph osmolality was calculated (x 1,000): H. pudibundus (25 per thousand, 8.1% kg H(2)O/mOsm x 10(3))>P. pandaliformis (2 per thousand, 9.2)>M. acanthurus (30 per thousand, 12.6)>P. pandaliformis (35 per thousand, 16.7)>D. pagei (7 per thousand, 60.4). Muscle slices submitted in vitro to a 30% osmotic challenge confirmed in vivo results. Thus, the estuarine/freshwater osmoregulators displayed a lower capacity to hold muscle tissue water than the marine osmoconformer, despite undergoing narrower variations in hemolymph osmolality. PMID:19844979

  13. Signal transduction, plasma membrane calcium movements, and pigment translocation in freshwater shrimp chromatophores.

    PubMed

    Milograna, Sarah Ribeiro; Bell, Fernanda Tinti; McNamara, John Campbell

    2010-11-01

    Crustacean color change results from the differential translocation of chromatophore pigments, regulated by neurosecretory peptides like red pigment concentrating hormone (RPCH) that, in the red ovarian chromatophores of the freshwater shrimp Macrobrachium olfersi, triggers pigment aggregation via increased cytosolic cGMP and Ca(2+) of both smooth endoplasmatic reticulum (SER) and extracellular origin. However, Ca(2+) movements during RPCH signaling and the mechanisms that regulate intracellular [Ca(2+)] are enigmatic. We investigate Ca(2+) transporters in the chromatophore plasma membrane and Ca(2+) movements that occur during RPCH signal transduction. Inhibition of the plasma membrane Ca(2+)-ATPase by La(3+) and indirect inhibition of the Na(+)/Ca(2+) exchanger by ouabain induce pigment aggregation, revealing a role for both in Ca(2+) extrusion. Ca(2+) channel blockade by La(3+) or Cd(2+) strongly inhibits slow-phase RPCH-triggered aggregation during which pigments disperse spontaneously. L-type Ca(2+) channel blockade by gabapentin markedly reduces rapid-phase translocation velocity; N- or P/Q-type blockade by ω-conotoxin MVIIC strongly inhibits RPCH-triggered aggregation and reduces velocity, effects revealing RPCH-signaled influx of extracellular Ca(2+). Plasma membrane depolarization, induced by increasing external K(+) from 5 to 50 mM, produces Ca(2+)-dependent pigment aggregation, whereas removal of K(+) from the perfusate causes pigment hyperdispersion, disclosing a clear correlation between membrane depolarization and pigment aggregation; K(+) channel blockade by Ba(2+) also partially inhibits RPCH action. We suggest that, during RPCH signal transduction, Ca(2+) released from the SER, together with K(+) channel closure, causes chromatophore membrane depolarization, leading to the opening of predominantly N- and/or P/Q-type voltage-gated Ca(2+) channels, and a Ca(2+)/cGMP cascade, resulting in pigment aggregation. PMID:20683865

  14. Deriving Freshwater Quality Criteria for Iron, Lead, Nickel, and Zinc for Protection of Aquatic Life in Malaysia

    PubMed Central

    Shuhaimi-Othman, M.; Nadzifah, Y.; Nur-Amalina, R.; Umirah, N. S.

    2012-01-01

    Freshwater quality criteria for iron (Fe), lead (Pb), nickel (Ni), and zinc (Zn) were developed with particular reference to aquatic biota in Malaysia, and based on USEPA's guidelines. Acute toxicity tests were performed on eight different freshwater domestic species in Malaysia which were Macrobrachium lanchesteri (prawn), two fish: Poecilia reticulata and Rasbora sumatrana, Melanoides tuberculata (snail), Stenocypris major (ostracod), Chironomus javanus (midge larvae), Nais elinguis (annelid), and Duttaphrynus melanostictus (tadpole) to determine 96 h LC50 values for Fe, Pb, Ni, and Zn. The final acute value (FAV) for Fe, Pb, Ni, and Zn were 74.5, 17.0, 165, and 304.9 μg L−1, respectively. Using an estimated acute-to-chronic ratio (ACR) of 8.3, the value for final chronic value (FCV) was derived. Based on FAV and FCV, a criterion maximum concentration (CMC) and a criterion continuous concentration (CCC) for Fe, Pb, Ni, and Zn that are 37.2, 8.5, 82.5, and 152.4 μg L−1 and 9.0, 2.0, 19.9, and 36.7 μg L−1, respectively, were derived. The results of this study provide useful data for deriving national or local water quality criteria for Fe, Pb, Ni, and Zn based on aquatic biota in Malaysia. Based on LC50 values, this study indicated that N. elinguis, M. lanchesteri, N. elinguis, and R. sumatrana were the most sensitive to Fe, Pb, Ni, and Zn, respectively. PMID:22919358

  15. 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.

  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. Resistance to the crayfish plague pathogen, Aphanomyces astaci, in two freshwater shrimps.

    PubMed

    Svoboda, J; Mrugała, A; Kozubíková-Balcarová, E; Kouba, A; Diéguez-Uribeondo, J; Petrusek, A

    2014-09-01

    Aphanomyces astaci, the causal agent of the crayfish plague, has recently been confirmed to infect also freshwater-inhabiting crabs. We experimentally tested the resistance of freshwater shrimps, another important decapod group inhabiting freshwaters, to this pathogen. We exposed individuals of two Asian shrimp species, Macrobrachium dayanum and Neocaridina davidi, to zoospores of the pathogen strain isolated from Procambarus clarkii, a known A. astaci carrier likely to get into contact with shrimps. The shrimps were kept in separate vessels up to seven weeks; exuviae and randomly chosen individuals were sampled throughout the experiment. Shrimp bodies and exuviae were tested for A. astaci presence by a species-specific quantitative PCR. The results were compared with amounts of A. astaci DNA in an inert substrate to distinguish potential pathogen growth in live specimens from persisting spores or environmental DNA attached to their surface. In contrast to susceptible crayfish Astacus astacus, we did not observe mortality of shrimps. The amount of detected pathogen DNA was decreasing steadily in the inert substrate, but it was still detectable several weeks after zoospore addition, which should be considered in studies relying on molecular detection of A. astaci. Probably due to moulting, the amount of A. astaci DNA was decreasing in N. davidi even faster than in the inert substrate. In contrast, high pathogen DNA levels were detected in some non-moulting individuals of M. dayanum, suggesting that A. astaci growth may be possible in tissues of this species. Further experiments are needed to test for the potential of long-term A. astaci persistence in freshwater shrimp populations. PMID:25064254

  18. 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

  19. Nitrogen and phosphorus cycling in shrimp ponds and the measures for sustainable management.

    PubMed

    Xia, L Z; Yang, L Z; Yan, M C

    2004-01-01

    Six ponds of age 3 were selected 45 km north from Suzhou in the Tailake region, and research conducted on nitrogen and phosphorus cycling in P. vannanmei (Penaeus vannanme) ponds and M. nipponense (Macrobrachium nipponense) hatchery ponds under normal management. Two treatments each had three replications. The results confirmed that feed was the major path of nitrogen and phosphorus input, each accounted for 61.24% (193.81 kg ha(-1)) and 81.08% (45.20 kg ha(-1)) of the total nitrogen and phosphorus input for P. vannanme ponds; the values for M. nipponense ponds were 43.93% (86.31 kg ha(-1)) and 57.67% (14.61 kg ha(-1)), respectively. Water pumped into ponds contributed on average 83.57 kg ha(-1) nitrogen and 8.48 kg ha(-1) phosphorus for P. vannanmei ponds, and 87.48 kg ha(-1) nitrogen and 7.00 kg ha(-1) phosphorus for M. nipponense hatchery ponds. Shrimp harvest recovered 102.81 kg ha(-1) nitrogen (32.94% of the total nitrogen input) and 7.94 kg ha(-1) phosphorus (14.23% of the total phosphorus input) for P. vannanme ponds; and 43.94 kg ha(-1) nitrogen and 4.46 kg ha(-1) phosphorus for M. nipponense hatchery ponds. The sum of nitrogen losses through volatilization, denitrification and sedimentation was 173.62 and 122.39 kg ha(-1), 54.86% and 62.29% of the total nitrogen input for P. vannanme ponds and M. nipponense hatchery ponds, respectively. Sediment accumulated 41.46 and 14.63 kg ha(-1) phosphorus, 74.37% and 64.85% of the total phosphorus input for P. vannanm ponds and M. nipponense hatchery ponds. Draining and seeping caused 40.06 kg ha(-1) nitrogen (12.66% of total nitrogen input) and 6.36 kg ha(-1) phosphorus (11.40% of total phosphorus input) loss to the surrounding water from P. vannanme ponds in 114 days; 30.14 kg ha(-1) nitrogen (15.34% of the total input) and 4.45 kg ha(-1) phosphorus (17.57% of the total input) to channel water from M. nipponense hatchery ponds in 87 days, respectively. Countermeasures for sustainable pond management include improving feeds and feeding, sediment treatments, machine aerating, chemicals with no pollution, and integrated fish-shrimp cultivation. Management of water resources for pond and methods to reduce nitrogen and phosphorus loading into surrounding water from drainage are elucidated. PMID:15499780

  20. 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

  1. Developing antibodies from cholinesterase derived from prokaryotic expression and testing their feasibility for detecting immunogen content in Daphnia magna.

    PubMed

    Liu, Hong-Cui; Yuan, Bing-Qiang; Li, Shao-Nan

    2016-02-01

    To yield cholinesterase (ChE) from prokaryotic expression, the ChE gene that belongs to Daphnia magna was amplified by reverse transcription-polymerase chain reaction (RT-PCR) using forward primer 5'-CCCYGGNGCSAT GATGTG-3' and reverse primer 5'-GYAAGTTRGCCCAATATCT-3'. To express the gene, one sequence of the amplified DNA, which was able to encode a putative protein containing two conserved carboxylesterase domains, was connected to the prokaryotic expression vector PET-29a(+). The recombinant vector was transformed into Escherichia coil BL21 (DE3). Protein expression was induced by isopropy-D-thiogalactoside. The expressed ChE was used as an immunogen to immunize BALB/c mice. The obtained antibodies were tested for their specificity towards crude enzymes from species such as Alona milleri, Macrobrachium nipponense, Bombyx mori, Chironomus kiiensis, Apis mellifera, Eisenia foetida, Brachydanio rerio, and Xenopus laevis. Results indicated that the antibodies had specificity suitable for detecting ChE in Daphnia magna. A type of indirect and non-competitive enzyme-linked immunosorbent assay (IN-ELISA) was used to test the immunoreactive content of ChE (ChE-IR) in Daphina magna. The detection limit of the IN-ELISA was found to be 14.5 ng/ml at an antiserum dilution of 1:22 000. Results from tests on Daphnia magna exposed to sublethal concentrations of triazophos indicated a maximal induction of 57.2% in terms of ChE-IR on the second day after the animals were exposed to a concentration of 2.10 μg/L triazophos. Testing on animals acclimatized to a temperature of 16 °C indicated that ChE-IR was induced by 16.9% compared with the ChE-IR content detected at 21 °C, and the rate of induction was 25.6% at 10 °C. The IN-ELISA was also used to test the stability of ChE-IR in collected samples. Repeated freezing and thawing had no influence on the outcome of the test. All these results suggest that the polyclonal antibodies developed against the recombinant ChE are as efficient as those developed against the native ChE in detecting ChE content in Daphnia magna. PMID:26834012

  2. Developing antibodies from cholinesterase derived from prokaryotic expression and testing their feasibility for detecting immunogen content in Daphnia magna *

    PubMed Central

    Liu, Hong-cui; Yuan, Bing-qiang; Li, Shao-nan

    2016-01-01

    To yield cholinesterase (ChE) from prokaryotic expression, the ChE gene that belongs to Daphnia magna was amplified by reverse transcription-polymerase chain reaction (RT-PCR) using forward primer 5'-CCCYGGNGCSAT GATGTG-3' and reverse primer 5'-GYAAGTTRGCCCAATATCT-3'. To express the gene, one sequence of the amplified DNA, which was able to encode a putative protein containing two conserved carboxylesterase domains, was connected to the prokaryotic expression vector PET-29a(+). The recombinant vector was transformed into Escherichia coil BL21 (DE3). Protein expression was induced by isopropy-D-thiogalactoside. The expressed ChE was used as an immunogen to immunize BALB/c mice. The obtained antibodies were tested for their specificity towards crude enzymes from species such as Alona milleri, Macrobrachium nipponense, Bombyx mori, Chironomus kiiensis, Apis mellifera, Eisenia foetida, Brachydanio rerio, and Xenopus laevis. Results indicated that the antibodies had specificity suitable for detecting ChE in Daphnia magna. A type of indirect and non-competitive enzyme-linked immunosorbent assay (IN-ELISA) was used to test the immunoreactive content of ChE (ChE-IR) in Daphina magna. The detection limit of the IN-ELISA was found to be 14.5 ng/ml at an antiserum dilution of 1:22 000. Results from tests on Daphnia magna exposed to sublethal concentrations of triazophos indicated a maximal induction of 57.2% in terms of ChE-IR on the second day after the animals were exposed to a concentration of 2.10 μg/L triazophos. Testing on animals acclimatized to a temperature of 16 °C indicated that ChE-IR was induced by 16.9% compared with the ChE-IR content detected at 21 °C, and the rate of induction was 25.6% at 10 °C. The IN-ELISA was also used to test the stability of ChE-IR in collected samples. Repeated freezing and thawing had no influence on the outcome of the test. All these results suggest that the polyclonal antibodies developed against the recombinant ChE are as efficient as those developed against the native ChE in detecting ChE content in Daphnia magna. PMID:26834012