Wangman, Pradit; Senapin, Saengchan; Chaivisuthangkura, Parin; Longyant, Siwaporn; Rukpratanporn, Sombat; Sithigorngul, Paisarn
The gene encoding the capsid protein of Macrobrachium rosenbergii nodavirus (MrNV) was cloned into pGEX-6P-1 expression vector and then transformed into the Escherichia coli strain BL21. After induction, capsid protein-glutathione-S-transferase (GST-MrNV; 64 kDa) was produced. The recombinant protein was separated using SDS-PAGE, excised from the gel, electro-eluted and then used for immunization for monoclonal antibody (MAb) production. Four MAbs specific to the capsid protein were selected and could be used to detect natural MrNV infections in M. rosenbergii by dot blotting, Western blotting and immunohistochemistry without cross-reaction with uninfected shrimp tissues or other common shrimp viruses. The detection sensitivity of the MAbs was 10 fmol µl-1 of the GST-MrNV, as determined using dot blotting. However, the sensitivity of the MAb on dot blotting with homogenate from naturally infected M. rosenbergii was approximately 200-fold lower than that of 1-step RT-PCR. Immunohistochemical analysis using these MAbs with infected shrimp tissues demonstrated staining in the muscles, nerve cord, gill, heart, loose connective tissue and inter-tubular tissue of the hepatopancreas. Although the positive reactions occurred in small focal areas, the immunoreactivity was clearly demonstrated. The MAbs targeted different epitopes of the capsid protein and will be used to develop a simple immunoassay strip test for rapid detection of MrNV. PMID:22436460
Longyant, Siwaporn; Senapin, Saengchan; Sanont, Sirijantra; Wangman, Pradit; Chaivisuthangkura, Parin; Rukpratanporn, Sombat; Sithigorngul, Paisarn
The capsid protein (CP) gene of extra small virus (XSV) expressed in Escherichia coli as a 42 kDa glutathione S-transferase (GST)-fusion protein (GST-XCP) or a 20 kDa His6-fusion protein (His6-XCP) were purified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), combined, and used to immunize Swiss mice to produce monoclonal antibodies (MAbs). Using dot blot, Western blot, and immunohistochemistry (IHC) methods, 4 MAbs specific to the XSV CP detected XSV in the freshwater prawn Macrobrachium rosenbergii without cross-reaction to host proteins or to proteins of Macrobrachium rosenbergii nodavirus (MrNV) or 5 of the most pathogenic viruses of penaeid shrimp. In dot blots, the combined MAbs could detect down to ~10 to 20 fmol µl-1 of purified GST-XCP protein, which was somewhat more sensitive compared to any single MAb. Used in conjunction with an MrNV-specific MAb, white tail disease (WTD) was diagnosed more effectively. However, the sensitivity at which the combined 4 MAbs detected XSV CP was 1000-fold lower than XSV RNA detected by RT-PCR. IHC analysis of M. rosenbergii tissue sections using the MAbs showed XSV infection to co-localize at variable loads with MrNV infection in heart and muscle cells as well as cells of connective tissues in the hepatopancreas. Since XSV histopathology remained prominent in tissues of some prawns in which MAb reactivity for MrNV was low compared to MAb reactivity for XSV, XSV might play some role in WTD severity. PMID:22832718
Lin, Feng; Liu, Li; Hao, Gui-Jie; Cao, Zheng; Sheng, Peng-Cheng; Wu, Ying-Lei; Shen, Jin-Yu
White coloration of the muscle of the giant river prawn (Macrobrachium rosenbergii) is a serious problem in China. The Macrobrachium rosenbergii Nodavirus (MrNV) has been confirmed to be the pathogen that causes this disorder. To develop a rapid, sensitive and specific technology for the detection of Macrobrachium rosenbergii Nodavirus isolated from China (MrNV-China), a reverse-transcription loop- mediated isothermal amplification assay combined with a lateral flow dipstick (RT-LAMP-LFD) assay method is described. A set of four primers and a labeled probe were designed specifically to recognize six distinct regions of the MrNV RNA2 gene. Results showed the sensitivity of the RT-LAMP-LFD assay was ten-times higher than the reverse-transcription loop-mediated isothermal amplification assay (RT-LAMP) with agarose gel electrophoresis. The assay was conducted with one-step amplification at 61°C in a single tube within 45 min. No product was generated from shrimps infected with other viruses, including DNA viruses (infectious hypodermal and hematopoietic necrosis virus (IHHNV); white spot syndrome virus (WSSV)) and RNA viruses (Taura syndrome virus (TSV); infectious myonecrosis virus (IMNV); yellow head virus (YHV)). Results were visualized by the LFD method. Therefore, the described rapid and sensitive assay is potentially useful for MrNV detection. PMID:25562958
Youngcharoen, Supak; Senapin, Saengchan; Lertwimol, Tareerat; Longyant, Siwaporn; Sithigorngul, Paisarn; Flegel, Timothy W; Chaivisuthangkura, Parin
Apoptosis is an essential immune response to protect invertebrates from virus infected cells. In shrimp, virus infection has been reported to induce apoptosis. Macrobrachium rosenbergii (Mr) was considered to be a disease-resistant host when compared to penaeid shrimps. Caspase-3 was classified as an executioner caspase which played a key role in virus-induced apoptosis. In this study, an effector caspase gene of M. rosenbergii (Mrcasp) was cloned and characterized. The open reading frame (ORF) of Mrcasp was 957 nucleotide encoding 318 amino acid with a deduced molecular mass of 35.87 kDa. RT-PCR analysis showed the presence of Mrcasp in all examined tissues. The phylogenetic tree indicated that Mrcasp was closely related with caspase 3 of shrimp. The functions of the Mrcasp, B2 and capsid proteins of M. rosenbergii nodavirus (MrNV) were assayed in Sf-9 cells. The results showed that Mrcasp induce apoptotic morphology cells; however, capsid protein of MrNV could inhibit apoptotic cells whereas B2 could neither induce nor inhibit apoptotic cells by DAPI staining. The protein interaction between Mrcasp and viral MrNV structure revealed that Mrcasp did not bind to B2 or capsid protein whereas B2 and capsid proteins could bind directly to each other. This study reported a novel sequence of a full-length Mrcasp and its functional studies indicated that Mrcasp could induce apoptotic cells. Our data is the first report demonstrating the direct protein-protein interaction between capsid protein and B2 protein of MrNV. PMID:25982399
Sahul Hameed, A S; Bonami, Jean-Robert
Macrobrachium rosenbergii is the most important cultured freshwater prawn in the world and it is now farmed on a large scale in many countries. Generally, freshwater prawn is considered to be tolerant to diseases but a disease of viral origin is responsible for severe mortalities in larval, post-larval and juvenile stages of prawn. This viral infection namely white tail disease (WTD) was reported in the island of Guadeloupe in 1995 and later in Martinique (FrenchWest Indies) in Taiwan, the People's Republic of China, India, Thailand, Australia and Malaysia. Two viruses, Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus-like particle (XSV) have been identified as causative agents of WTD. MrNV is a small icosahedral non-enveloped particle, 26-27 nm in diameter, identified in the cytoplasm of connective cells. XSV is also an icosahedral virus and 15 nm in diameter. Clinical signs observed in the infected animals include lethargy, opaqueness of the abdominal muscle, degeneration of the telson and uropods, and up to 100 % within 4 days. The available diagnostic methods to detect WTD include RT-PCR, dot-blot hybridization, in situ hybridization and ELISA. In experimental infection, these viruses caused 100 % mortality in post-larvae but failed to cause mortality in adult prawns. The reported hosts for these viruses include marine shrimp, Artemia and aquatic insects. Experiments were carried out to determine the possibility of vertical transmission of MrNV and XSV in M. rosenbergii. The results indicate that WTD may be transferred from infected brooders to their offspring during spawning. Replication of MrNV and XSV was investigated in apparently healthy C6/36 Aedes albopictus and SSN-1 cell lines. The results revealed that C6/36 and SSN-1cells were susceptible to these viruses. No work has been carried out on control and prevention of WTD and dsRNA against protein B2 produced RNAi that was able to functionally prevent and reduce mortality in WTD-infected redclaw crayfish. PMID:23997437
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...
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.
Warachin Gangnonngiw; Kesinee Laisutisan; Siriporn Sriurairatana; Saengchan Senapin; Niti Chuchird; Chalor Limsuwan; Parin Chaivisuthangkura; Timothy W. Flegel
Field specimens of post-larvae of the giant freshwater prawn (Macrobrachium rosenbergii) from Thailand showed hepatopancreatic tubule epithelial cells that contained central, eosinophilic inclusions within enlarged nuclei and marginated chromatin. These inclusions resembled those produced by some baculoviruses prior to formation of occlusion bodies that enclose virions in a polyhedrin protein matrix. By electron microscopy, the intranuclear inclusions contained bacilliform, enveloped
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...
Hung-Hung Sung; Shu-Fen Hwang; Fu-Ming Tasi
The virulence of two Aeromonas strains (A. veronii and A. caviae) isolated from the hepatopancreas of apparently healthy giant freshwater prawns (Macrobrachium rosenbergii) was compared using a challenge by injections. For the A. veronii strain, challenge with 3.7 × 105 cells\\/g of body weight led to 100% mortality; for the A. caviae strain, 3.8 × 106 cells\\/g produced 100% mortality.
Llobrera, Alcestis Trillo
in this study. The shrimp were held in holding facilities at the Texas A&M University, Department of Veterinary Microbiology and Parasitology, Aquatic Animal Medicine Laboratory. Ralston Purina Experimental Marine Ration No. 25 (Checkerboard Square, St... for the degree of MASTER OF SCIENCE August 1980 Major Subject: Veterinary Microbiology and Parasitology ANTIBIOTIC ADMINISTRATION BY OSMOTIC INFILTRATION IN THE FRESHWATER SHRIMP, MACROBRACHIUM ROSENBERGII A Thesis by ALCESTIS TRILLO LLOBRERA Approved...
VanMaurik, Lauren N.; Wortham, Jennifer L.
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
Shinn-Pyng Yeh; Chun-Hung Liu; Tzeng-Gan Sung; Pai-Po Lee; Winton Cheng
Giant freshwater prawns, Macrobrachium rosenbergii (17.9±2.7 g), exposed to different concentrations of saponin at 0, 0.3, 0.6, 0.9 and 1.2 mg l?1 for 168 h were examined for osmolality, electrolyte levels, oxyhemocyanin, protein levels, acid-base balance status, total hemocyte count (THC), phenoloxidase activity, and respiratory bursts. Hemolymph oxyhemocyanin, protein, and pO2 were inversely related to the saponin concentration. Hemolymph oxyhemocyanin, protein, pO2, pCO2,
Chin-Chyuan Chang; Ming-Di Hung; Winton Cheng
In this study, we determined the effects of norepinephrine (NE) on immunity and the pathway of its function in the freshwater giant prawn, Macrobrachium rosenbergii. The total hemocyte count (THC), differential hemocyte count (DHC), phenoloxidase activity, respiratory bursts, superoxide dismutase (SOD) activity, phagocytic activity, and clearance efficiency in response to the pathogen, Lactococcus garvieae, were measured when the freshwater giant
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...
Marcy N. Wilder; Wei-Jun Yang; Do Thi Thanh Huong; Masachika Maeda
The giant freshwater prawn, Macrobrachium rosenbergii , is a commercially important species of crustacean cultured extensively throughout Southeast Asia. In Vietnam, where Japan International Research Center for Agricultural Sciences (JIRCAS) is currently implementing a comprehensive project entitled \\
Huang, Ying; Ren, Qian
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
Gangnonngiw, Warachin; Laisutisan, Kesinee; Sriurairatana, Siriporn; Senapin, Saengchan; Chuchird, Niti; Limsuwan, Chalor; Chaivisuthangkura, Parin; Flegel, Timothy W
Field specimens of post-larvae of the giant freshwater prawn (Macrobrachium rosenbergii) from Thailand showed hepatopancreatic tubule epithelial cells that contained central, eosinophilic inclusions within enlarged nuclei and marginated chromatin. These inclusions resembled those produced by some baculoviruses prior to formation of occlusion bodies that enclose virions in a polyhedrin protein matrix. By electron microscopy, the intranuclear inclusions contained bacilliform, enveloped virions (approximately 327+/-29nmx87+/-12nm) with evenly dense, linear nucleocapsids surrounded by trilaminar envelopes with lateral pockets containing nucleoproteinic filaments. In some cases, these were accompanied by moderately electron dense, spherical particles of approximately 20nm diameter resembling polyhedrin subunits of occlusion bodies (OB) of a bacilliform virus of the black tiger shrimp Penaeus monodon, previously reported from Thailand and called monodon baculovirus (MBV). It is currently listed by the International Committee on Taxonomy of viruses as Penaeus monodon nucleopolyhedrovirus (PemoNPV). Two polymerase chain reaction (PCR) assays for MBV gave positive results with DNA extracts prepared from M. rosenbergii samples using the hot phenol technique. One of these assays targeted the polyhedrin gene of MBV to which the resulting amplicon showed 100% sequence identity. Presence of the Penaeus monodon virus polyhedrin gene was confirmed by in situ hybridization assays and by positive immunohistochemical reactions in one sample batch. The data revealed that MBV can be found but may rarely produce polyhedrin occlusion bodies in M. rosenbergii. PMID:19963025
Chowdhury, Labrechai Mog; Gireesh-Babu, P; Pavan-Kumar, A; Suresh Babu, P P; Chaudhari, Aparna
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
Revathi, Peranandam; Vasanthi, Lourduraj Arockia; Munuswamy, Natesan
In this investigation, effect of cadmium chloride (25 ?g/l) on oogenesis of freshwater prawn, Macrobrachium rosenbergii was studied. In vivo experiments were performed with both intact and eyestalk ablated prawns. The intact, cadmium-exposed prawns exhibited decrease in Gonado Somatic Index (GSI) and ovarian development compared to controls. Whereas, ablated treated ovary showed reduction of yolk material and oocyte membrane thickness at the end of 15 days exposure. Interestingly, the control prawn showed normal cellular architecture of gills, hepatopancreas and ovary with mature oocytes. But, the gills of treated prawns showed lamellar hypertrophy, cuticular dystrophy and irregular arrangement of epithelial cells. Hepatopancreas showed reduction in both tubular diameter and basement membrane thickness. Conspicuously, ovary showed hypertrophied primary oocytes with more vacuoles in intact-treated group. Cadmium had increased gonad inhibiting hormone (GIH) secretion and decreased gonad stimulating hormone (GSH) release as evident with the retardation of gonadal maturation in the intact prawns. PMID:21296420
Tsuyoshi Ohira; Naoaki Tsutsui; Hiromichi Nagasawa; Marcy N. Wilder
Crustacean hyperglycemic hormone (CHH) is released from the X-organ\\/sinus gland complex located in the eyestalks, and regulates glucose levels in the hemolymph. In the giant freshwater prawn ( Macrobrachium rosenbergii ), two cDNAs encoding different CHH molecules were previously cloned by other workers. One of these (Mar-CHH-2) was expressed only in the eyestalks, whereas the other (Mar-CHH-L) was expressed in
Shu-Hwa Chen; Chung-Yen Lin; Ching Ming Kuo
A full-length chh cDNA was cloned from the eyestalk of Macrobrachium rosenbergii. The 991-bp cDNA contains an open reading frame of 408 bp that encodes the prepro-CHH. The tissue-specific expression pattern was examined by reverse transcriptase-polymerase chain reaction. Positive signals were detected in the eyestalk, heart, gills, antennal glands, and thoracic ganglion but not in muscle and hepatopancreas. However, two
T Ventura; E D Aflalo; S Weil; K Kashkush; A Sagi
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
Na Li; Yunlong Zhao; Jian Yang
The aim of the present study was to evaluate the potential utility of enzyme parameters as indicators of water-borne copper\\u000a (Cu2+) contamination in the giant freshwater prawn Macrobrachium rosenbergii. Activities of the digestive enzymes of tryptase, pepsin, cellulase, amylase, and metabolic enzymes of alkaline phosphatase\\u000a (AKP), acid phosphatase (ACP), superoxide dismutase (SOD) and glutathione-S-transferase (GST) were measured in the hepatopancreas
Shi, Yan-Ru; Jin, Min; Ma, Fu-Tong; Huang, Ying; Huang, Xin; Feng, Jin-Ling; Zhao, Ling-Ling; Chen, Yi-Hong; Ren, Qian
Relish is an NF-kB transcription factor involved in immune-deficiency (IMD) signal pathway. In this study, a Relish gene (MrRelish) was identified from Macrobrachium rosenbergii. The full length of MrRelish comprises 5072 bp, including a 3510 bp open reading frame encoding a 1169 bp amino acid protein. MrRelish contains a Rel homology domain (RHD), a nucleus localization signal, an I?B-like domain (6 ankyrin repeats), and a death domain. Phylogenetic analysis showed that MrRelish and other Relish from crustaceans belong to one group. MrRelish was expressed in all detected tissues, with the highest expression level in hemocytes and intestines. MrRelish was also upregulated in hepatopancreas at 6?h after Vibrio anguillarum challenge. The over-expression of MrRelish could induce the expression of antimicrobial peptides (AMPs), such as Drosophila Metchnikowin (Mtk), Attacin (Atta), Drosomycin (Drs), and Cecropin (CecA) and shrimp Penaeidin (Pen4). The RNAi of MrRelish in gills showed that the expression of crustin (cru) 2, Cru5, Cru8, lysozyme (Lyso) 1, and Lyso2 was inhibited. However, the expression of anti-lipopolysaccharide factor (ALF) 1 and ALF3 did not change when MrRelish was knocked down. These results indicate that MrRelish may play an important role in innate immune defense against V. anguillarum in M. rosenbergii. PMID:26026243
Law, A.T.; Yeo, M.E. [Universiti Kolej Terengganu (UPM), Darul Iman (Malaysia)] [Universiti Kolej Terengganu (UPM), Darul Iman (Malaysia)
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.
Arockiaraj, Jesu; Gnanam, Annie J; Palanisamy, Rajesh; Bhatt, Prasanth; Kumaresan, Venkatesh; Chaurasia, Mukesh Kumar; Pasupuleti, Mukesh; Ramaswamy, Harikrishnan; Arasu, Abirami; Sathyamoorthi, Akila
Glutathione S-transferases play an important role in cellular detoxification and may have evolved to protect cells against reactive oxygen metabolites. In this study, we report the molecular characterization of glutathione s-transferase-theta (GST-?) from freshwater prawn Macrobrachium rosenbergii. A full length cDNA of GSTT (1417 base pairs) was isolated and characterized bioinformatically. Exposure to virus (white spot syndrome baculovirus or M. rosenbergii nodovirus), bacteria (Aeromonas hydrophila or Vibrio harveyi) or heavy metals (cadmium or lead) significantly increased the expression of GSTT (P<0.05) in hepatopancreas. Recombinant GST-? with monochlorobimane substrate had an optimum activity at pH7.5 and 35 °C. Furthermore recombinant GST-? activity was abolished by the denaturants triton X-100, Gua-HCl, Gua-thiocyanate, SDS and urea in a dose-dependent manner. Overall, the results suggest a potential role for M. rosenbergii GST-? in detoxification and possibly conferring immune protection. PMID:24879918
Jung, Hyungtaek; Lyons, Russell E.; Dinh, Hung; Hurwood, David A.; McWilliam, Sean; Mather, Peter B.
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
Zhang, Xiao-Wen; Wang, Xian-Wei; Huang, Ying; Hui, Kai-Min; Shi, Yan-Ru; Wang, Wen; Ren, Qian
Ficolins, a kind of lectin containing collagen-like and fibrinogen-related domains (FReDs, also known as FBG or FREP), are involved in the first line of host defense against pathogens. In this study, two ficolins, namely, MrFico1 and MrFico2, from the giant freshwater prawn Macrobrachium rosenbergii were identified. In contrast to other ficolins, these two ficolins have no collagen-like domain, but such ficolins contain a coiled region and a FReD domain. Phylogenetic analysis showed that MrFico1 and MrFico2, together with two ficolin-like proteins from Pacifastacus leniusculus, belonged to one group. Quantitative RT-PCR (qRT-PCR) showed that both MrFico1 and MrFico2 were expressed in hepatopancreas, stomach and intestine, with the highest expression in stomach for MrFico1, compared to the highest expression in hepatopancreas for MrFico2. qRT-PCR analysis also showed that MrFico1 was obviously upregulated upon Vibrio anguillarium challenge, while MrFico2 was upregulated after challenged by V. anguillarium or white spot syndrome virus. Bacterium-binding experiment showed that MrFico1 and MrFico2 could bind to different microbes, and sugar-binding assay revealed that these two ficolins could also bind to lipopolysaccharide and peptidoglycan, the glycoconjugates of bacteria surface. Moreover, these two ficolins could agglutinate bacteria in a calcium-dependent manner, and the results of bacteria clearance experiment showed that both ficolins could facilitate the clearance of injected bacteria in the prawn. Our results suggested that MrFico1 and MrFico2 may function as pattern-recognition receptors in the immune system of M. rosenbergii. PMID:24462836
Hsu, Pei-I; Liu, Chun-Hung; Tseng, Deng-Yu; Lee, Pai-Po; Cheng, Winton
Expression of peroxinectin cDNA was determined from haemocytes of giant freshwater prawn Macrobrachium rosenbergii using oligonucleotide primers and reverse transcription polymerase chain reaction (RT-PCR) based on the peroxinectin sequence of white shrimp Litopenaeus vannamei, tiger shrimp Penaeus monodon, and freshwater crayfish Pacifastacus leniusculus. The peroxinectin of M. rosenbergii was constitutively expressed. Analysis of the nucleotide sequence revealed that the cDNA clone has an open reading frame of 2,403 bp encoding a protein of 801 amino acids including a 20 amino acid signal peptide. The calculated molecular mass of the mature protein (781 amino acids) was 88.7 kDa with an estimated pI of 6.8. A putative peroxidase domain and a putative integrin-binding motif, KGD (Lys-Gly-Asp) were observed in prawn peroxinectin at the C-terminal. Sequence comparison showed that peroxinectin deduced amino acid of M. rosenbergii had an overall similarity of 62%, 64%, and 66% to that of P. leniusculus, P. monodon, and L. vannamei, respectively. Quantitative real-time PCR analysis showed that peroxinectin transcript in haemocyte of M. rosenbergii decreased significantly after 3, 6 and 12h injection with Lactococcus garvieae. PMID:16377210
Liu, Chun-Hung; Yeh, Shinn-Pyng; Hsu, Pei-Yi; Cheng, Winton
Peroxinectin (PE) gene expressions were determined using real-time PCR in the giant freshwater prawn Macrobrachium rosenbergii based on moulting; prawns were fed diets containing different concentrations of sodium alginate, and were exposed to different concentrations of copper sulphate, benzalkonium chloride (BKC), and trichlorfon. Results showed that PE mRNA expression of prawns was the highest in stage A, significantly decreased in stage B, and reached the lowest level in stages D0/D1. The PE transcript was significantly higher in prawns fed the 1.0 gkg(-1) sodium alginate-containing diet than those fed the 2.0 gkg(-1) sodium alginate-containing diet and those fed the control diet. PE transcripts significantly decreased in prawns exposed to 0.1-0.4 mgL(-1) copper sulphate after 96 h, 0.3-1.0 mgL(-1) BKC after 96 h, and 0.2-0.4 mgL(-1) trichlorfon after 48 h. It was concluded that the status of PE gene expression was seriously affected by the moult cycle, immunostimulant, and chemotherapeutants. PMID:17056274
Winton Cheng; Chun-Hung Liu; Jung-Ping Hsu; Jiann-Chu Chen
Giant freshwater prawns Macrobrachium rosenbergii (14–19g) were challenged withEnterococcus (3×105cfu prawn?1) previously incubated in TSB medium for 24h, then placed in water having concentrations of dissolved oxygen (DO) at 7·75, 4·75, 2·75 and 1·75mg l?1. Onset of mortality occurred after 6h exposure to 1·75mg l?1 DO, and after 12h exposure to 2·75mg l?1 DO. Cumulative mortality of prawns at 1·75mg
S. J. Funge-Smith; A. C. Taylor; J. Whitley; J. H. Brown
Macrobrachium rosenbergii is able to exert a high degree of control over the osmotic pressure and the ionic concentration of its haemolymph when exposed to a wide range of salinity between fresh water and its isosmotic point. Sodium and chloride ions were regulated at almost identical concentrations over most of the salinity range tested. Calcium and potassium ions were hyperregulated
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
Yadzir, Zailatul Hani Mohamad; Misnan, Rosmilah; Abdullah, Noormalin; Bakhtiar, Faizal; Arip, Masita; Murad, Shahnaz
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
Du, Jie; Zhu, Huanxi; Ren, Qian; Liu, Peng; Chen, Jing; Xiu, Yunji; Yao, Wei; Meng, Qingguo; Gu, Wei; Wang, Wen
Flow cytometry provides rapid and reproducible methods for analyzing crustacean cellular immune responses to pathogens. We used this method to investigate the hemocytes sub-populations of freshwater prawn Macrobrachium rosenbergii and their immune responses to a novel pathogen spiroplasma MR-1008. M. rosenbergii inoculated with 100 ?l spiroplasma strain MR-1008 in logarithmic phase (10(8) spiroplasmas ml(-1)) were examined for total hemocytes count (THC) and changes in differential involvement of hemocytes sub-populations during 1-28 d after inoculation. The results showed that THC was dramatically lowered 1 d after inoculation, and it obviously increased at the 5 d after inoculation; thereafter, a high level of THC was maintained to 15 d. Three morphologically distinct hemocytes sub-populations including granular cells (GC), semigranular cells (SGC) and hyaline cells (HC) could be identified by flow cytometry, and the proportions of the 3 kinds of cell categories varied obviously during the infection of spiroplasma suggesting differential involvement according to the pathogen. The flow cytometry used in this study confirmed that the semigranular cells were the main hemocytes involved in the cellular defense against spiroplasma in the M. rosenbergii. PMID:22842149
Wang, Tianfang; Zhao, Min; Elizur, Abigail; Sretarugsa, Prapee; Cummins, Scott F.; Sobhon, Prasert
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
Liu, Chun-Hung; Chang, Chin-Chyuan; Chiu, Yun-Chih; Cheng, Winton; Yeh, Maw-Sheng
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
Chang, Chin-Chyuan; Rahmawaty, Atiek; Chang, Zhong-Wen
Trichlorfon is an organophosphorus (OP) insecticide that is used as an agriculture pesticide to destroy insects, a human medicine to combat internal parasites, and an ectoparasiticide in the livestock and aquaculture industries, but which has caused aquatic toxicity in the prawn industry. The aim of this study was to investigate the effects of trichlorfon on molecular and enzymatic processes of the immunological response of the giant freshwater prawn, Macrobrachium rosenbergii, at 0, 0.2, and 0.4mgL(-1) with 0, 3, 6, 12, and 24h of exposure. The total hemocyte count (THC), respiratory bursts (RBs), phenoloxidase (PO) activity, and superoxide dismutase (SOD) activity were examined to evaluate immunological responses and oxidative stress. Results showed that THCs of the prawn exposed to trichlorfon at both concentrations (0.2 and 0.4mgL(-1)) had increased after 12 and 24h; SOD and PO activities had significantly increased at 3h, whereas production of RBs had dramatically increased as oxidative stress at each sampling time after exposure to trichlorfon compared to the control. A potential biomarker of OPs, acetylcholinesterase (AChE) revealed a significant decrease after exposure for 6h, and showed a time-dependent tendency. Immune gene expressions, including prophenoloxidase (proPO), the lipopolysaccharide- and ?-1,3-glucan-binding protein (LGBP), peroxinectin (PE), ?2-macroglubulin (?2M), transglutaminase (TG), and copper, zinc (Cu,Zn)-SOD, of prawns exposed to trichlorfon at 0, 0.2, and 0.4mgL(-1) for 0, 6, and 24h were further evaluated. Expressions of all of the immune genes significantly decreased when prawns were exposed to 0.4mgL(-1) trichlorfon for 24h, and among them, an increase in SOD expression was seen after exposure to 0.4mgL(-1) for 6h. Prawns exposed to trichlorfon within 24h exhibited the decrease of circulating hemocytes, and also the induction of oxidative stress, which caused subsequent damage to DNA formation of immune genes. From these results, we concluded that immunological responses and immune gene expressions of prawn exposed to trichlorfon at 0.4mgL(-1) for 24h were perturbed, thus causing a deficiency in immunity and subsequent increased susceptibility to pathogen infections. PMID:23340335
Shieh-Tsung Chiu; Shu-Ling Hsieh; Shinn-Pyng Yeh; Shun-Ji Jian; Winton Cheng; Chun-Hung Liu
The effects of inorganic selenium (Se) (sodium selenate, SSe) and organic selenium (seleno-l-methionine, MSe) supplementation on the immune response, antioxidant status, and disease resistance of the giant freshwater prawn, Macrobrachium rosenbergii, were studied. Five experimental diets, including a control diet (without Se enrichment), 0.5 mg (kg diet)?1 of MSe, 1 mg (kg diet)?1 of MSe, 0.5 mg (kg diet)?1 of SSe, and 1 mg
Harikrishnan, Ramasamy; Balasundaram, Chellam; Jawahar, Sundaram; Heo, Moon-Soo
The effect of Withania somnifera extract supplementation diets on innate immune response in giant freshwater prawn Macrobrachium rosenbergii (de Man) against Aeromonas hydrophila was investigated. The bacterial clearance efficiency significantly increased in prawn fed with 0.1% and 1.0% doses of W. somnifera supplementation diet against pathogen from weeks 1-4 as compared to the control. The innate immune parameters such as, phenoloxidase activity, superoxide anion level, superoxide dismutase activity, nitrate, and nitrite concentrations were significantly enhanced in prawn fed with 0.1% and 1.0% doses of W. somnifera supplementation diet from weeks 1-4 against pathogen. The total hemocyte counts (THC) significantly increased in prawn fed with 0.1% and 1.0% doses diet from weeks 1-4 against pathogen as compared to the control. These results strongly suggested that administration of W. somnifera through supplementation diet positively enhances the innate immune system and enhanced survival rate in M. rosenbergii against A. hydrophila infection. PMID:22118967
Ventura, T; Aflalo, E D; Weil, S; Kashkush, K; Sagi, A
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
Chang, Chin-Chyuan; Hung, Ming-Di; Cheng, Winton
In this study, we determined the effects of norepinephrine (NE) on immunity and the pathway of its function in the freshwater giant prawn, Macrobrachium rosenbergii. The total hemocyte count (THC), differential hemocyte count (DHC), phenoloxidase activity, respiratory bursts, superoxide dismutase (SOD) activity, phagocytic activity, and clearance efficiency in response to the pathogen, Lactococcus garvieae, were measured when the freshwater giant prawn, M. rosenbergii (16.2±2.1 g) was individually injected with saline or NE at 0.5, 5.0, and 50.0 pmol prawn(-1). Results showed that semi-granular cells, respiratory bursts and phagocytic activity at 2 h, PO activity and clearance efficiency from 2 to 4 h, THC at 8 h, and SOD activity from 4 to 8 h significantly decreased, but hyaline cells at 2 h, and respiratory bursts at 8 h had significantly increased after injection of NE at 50.0 pmol prawn(-1). In prawns that had received 5.0 pmol NE prawn(-1), the PO activity had decreased at 2 h, SOD activity at 8 h, and the clearance efficiency at 2 h. PO activity had decreased at 2 h after prawns had received 0.5 pmol NE prawn(-1). All of the immune parameters had returned to control values by 16 h after receiving NE. However, no significant differences were observed in the granular cells during the experimental period. An injection of NE also significantly increased the mortality of prawns challenged with L. garvieae, which appeared to be dose dependent. In another experiment, NE co-injected with prazosin, metoprolol, or propranolol significantly decreased the mortality of challenged prawns, especially when co-injected with prazosin and metoprolol. These results suggest that stress-inducing NE suppresses the immune system, which in turn promotes the susceptibility of M. rosenbergii to L. garvieae via both ?1- and ?1-adrenergic receptors. PMID:21295067
Cheng, Winton; Wang, Ching-Hsien; Chen, Jiann-Chu
Addition of benzalkonium chloride (BKC) at 0, 0.3, 0.6 and 1.0 mg l(-1) to tryptic soy broth (TSB) had no effect on growth of Lactococcus garvieae, a bacterial pathogen of the giant freshwater prawn Macrobrachium rosenbergii. However, injection of the cultured cells into prawns at a dose of 4 x 10(6) colony-forming units (cfu) prawn(-1) resulted in significantly higher mortality at 120 h (p < 0.05) in prawns injected with cells grown in the absence of BKC than in prawns injected with cells grown in the presence of BKC. In other experiments, prawns were injected with TSB-grown L. garvieae (4 x 10(6) and 3 x 10(5) cfu prawn(-1)) and then held in water containing BKC at 0, 0.3, 0.6 and 1.0 mg l(-1). After 120 h, mortality was significantly higher in all the BKC treatments than in the control without BKC. Prawns showed no significant differences in total hemocyte count (THC) or differential hemocyte count (DHC) amongst treatment and control groups. However, 96 h exposure to 0.3 mg l(-1) BKC or more resulted in a decrease in phenoloxidase activity and an increase in respiratory burst to levels considered to be cytoxic. In summary, exposure of L. garvieae to BKC at 0.3 mg l(-1) or more decreased its virulence to M. rosenbergii, while exposure of M. rosenbergii to BKC at 0.3 mg l(-1) or more increased its susceptibility to L. garvieae infection. PMID:12691193
Ding, Zheng-Feng; Ren, Jie; Tan, Jing-Min; Wang, Zheng; Yin, Shao-Wu; Huang, Ying; Huang, Xin; Wang, Wen; Lan, Jiang-Feng; Ren, Qian
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
Huang, Xin; Huang, Ying; Shi, Yan-Ru; Ren, Qian; Wang, Wen
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
Barman, Hirak Kumar; Patra, Swagat Kumar; Das, Varsha; Mohapatra, Shibani Dutta; Jayasankar, Pallipuram; Mohapatra, Chinmayee; Mohanta, Ramya; Panda, Rudra Prasanna; Rath, Surya Narayan
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
Hui, Kai-Min; Hao, Fang-Yuan; Li, Wen; Zhang, Zhao; Zhang, Chi-Yu; Wang, Wen; Ren, Qian
Glutathione S-transferases (GSTs) are essential components of the cellular detoxification system because of their capability to protect organisms against the toxicity of reactive oxygen species (ROSs). Four different GSTs (MrMuGST1-MrMuGST4) showing similarities with Mu-type GSTs were cloned from the hepatopancreas of Macrobrachium rosenbergii. These four GSTs have 219, 216, 218 and 219 amino acids in length, respectively. MrMuGST1-MrMuGST4 proteins all have a G-site in the N-terminus and an H-site in the C-terminus. Phylogenetic analysis reveals that four Mu-type GSTs are classified into two different clades (MrMuGST2 one clade; MrMuGST1, MrMuGST3 and MrMuGST4 other clades). Nonetheless, no site under positive selection was detected but rapid evolution was found in the few of MuGST genes. Reverse transcription-polymerase chain reaction (RT-PCR) results showed that MrMuGST1 and MrMuGST2 transcripts were expressed in all detected tissues, however, MrMuGST3 and MrMuGST4 were just mainly expressed in hepatopancreas and intestines. Quantitative RT-PCR analysis showed that MrMuGST1 and MrMuGST2 were down-regulated upon Vibrio anguillarum challenge, whereas MrMuGST3 and MrMuGST4 were quickly up-regulated 2 h after the Vibrio challenge. Our results imply that different Mu-type GSTs may respond to Vibrio challenge with different manners. PMID:23727284
Ye, Ting; Huang, Xin; Wang, Xian-Wei; Shi, Yan-Ru; Hui, Kai-Min; Ren, Qian
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
Chang, Chin-Chyuan; Tan, Hui-Ching; Cheng, Winton
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
Nguyen Thanh, Hai; Zhao, Liangjie; Liu, Qigen
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
Iketani, Gabriel; Pimentel, Luciana; Silva-Oliveira, Glaúcia; Maciel, Cristiana; Valenti, Wagner; Schneider, Horacio; Sampaio, Iracilda
The giant river prawn, Macrobrachium cf. rosenbergii, is one of the most cultivated freshwater prawns in the world and has been introduced into more than 40 countries. In some countries, this prawn is considered an invasive species that requires close monitoring. Recent changes in the taxonomy of this species (separation of M. rosenbergii and M. dacqueti) require a re-evaluation of introduced taxa. In this work, molecular analyses were used to determine which of these two species was introduced into Brazil and to establish the geographic origin of the introduced populations that have invaded Amazonian coastal waters. The species introduced into Brazil was M. dacqueti through two introduction events involving prawns originating from Vietnam and either Bangladesh or Thailand. These origins differ from historical reports of the introductions and underline the need to confirm the origin of other exotic populations around the world. The invading populations in Amazonia require monitoring not only because the biodiversity of this region may be affected by the introduction, but also because admixture of different native haplotypes can increase the genetic variability and the likelihood of persistence of the invading species in new habitats. PMID:21637558
Liu, Chia-Chen; Chung, Chien-Pang; Lin, Chang-Yi; Sung, Hung-Hung
In this study, a 780-bp full-length cDNA encoding Macrobrachium rosenbergii anti-lipopolysaccharide factor (MrALF) from hemocytes was cloned and identified. The ALF isoform exhibited immune activities, and its concentration in hemolymph was determined. An in vivo expression study showed that the ALF mRNA level of hemocytes could be induced by lipopolysaccharides (LPSs) in an exposure time-dependent manner. Purified recombinant MrALF (rMrALF) expressed in the yeast Pichia pastoris SMD1168 eukaryotic protein expression system demonstrated antibacterial activity against the Gram-negative prawn pathogen Aeromonas hydrophila (minimum inhibitory concentration (MIC)=0.806?M, minimum bactericidal concentration (MBC)=1.606?M) but not the Gram-positive pathogen Lactococcus garvieae exposed to 25.696?M of rALF. However, rMrALF can bind to Gram-negative and -positive bacteria. An in vivo expression study demonstrated that the ALF concentrations in prawn hemocytes and plasma were 0.176?M and 0.168?M, respectively; following LPS treatment for 6h, the corresponding concentrations were 0.133?M in hemocytes and 0.272?M in plasma. Furthermore, the percentage of hemocytes phagocytosing bacteria cells was higher in hemoyctes previously treated with MrALF than those treated with sterile medium. These results suggest that in the innate immune response of M. rosenbergii, the MrALF from hemocytes may play an opsonin during a bacterial invasion. PMID:24333685
Siangcham, Tanapan; Tinikul, Yotsawan; Poljaroen, Jaruwan; Sroyraya, Morakot; Changklungmoa, Narin; Phoungpetchara, Ittipon; Kankuan, Wilairat; Sumpownon, Chanudporn; Wanichanon, Chaitip; Hanna, Peter J; Sobhon, Prasert
Neurotransmitters and neurohormones are agents that control gonad maturation in decapod crustaceans. Of these, serotonin (5-HT) and dopamine (DA) are neurotransmitters with known antagonist roles in female reproduction, whilst gonadotropin-releasing hormones (GnRHs) and corazonin (Crz) are neurohormones that exercise both positive and negative controls in some invertebrates. However, the effects of these agents on the androgenic gland (AG), which controls testicular maturation and male sex development in decapods, via insulin-like androgenic gland hormone (IAG), are unknown. Therefore, we set out to assay the effects of 5-HT, DA, l-GnRH-III, oct-GnRH and Crz, on the AG of small male Macrobrachium rosenbergii (Mr), using histological studies, a BrdU proliferative cell assay, immunofluorescence of Mr-IAG, and ELISA of Mr-IAG. The results showed stimulatory effects by 5-HT and l-GnRH-III through significant increases in AG size, proliferation of AG cells, and Mr-IAG production (P<0.05). In contrast, DA and Crz caused inhibitory effects on the AG through significant decreases in AG size, proliferation of AG cells, and Mr-IAG production (P<0.05). Moreover, the prawns treated with Crz died before day 16 of the experimental period. We propose that 5-HT and certain GnRHs can be now used to stimulate reproduction in male M. rosenbergii, as they induce increases in AG and testicular size, IAG production, and spermatogenesis. The mechanisms by which these occur are part of our on-going research. PMID:23867230
Chiu, Shieh-Tsung; Hsieh, Shu-Ling; Yeh, Shinn-Pyng; Jian, Shun-Ji; Cheng, Winton; Liu, Chun-Hung
The effects of inorganic selenium (Se) (sodium selenate, SSe) and organic selenium (seleno-l-methionine, MSe) supplementation on the immune response, antioxidant status, and disease resistance of the giant freshwater prawn, Macrobrachium rosenbergii, were studied. Five experimental diets, including a control diet (without Se enrichment), 0.5 mg (kg diet)(-1) of MSe, 1 mg (kg diet)(-1) of MSe, 0.5 mg (kg diet)(-1) of SSe, and 1 mg (kg diet)(-1) of SSe, were used. After 75 days of culture, prawn fed the Se-enriched diets had lower mortality compared to that of prawn fed the control diet after being challenged by the pathogen, Debaryomyces hansenii. No significant differences in the total hemocyte count, superoxide dismutase activity, or clearance efficiency of prawn were recorded among the control and treated groups. Significantly increased phenoloxidase and phagocytic activities in prawn fed the Se-enriched diets were found compared to the controls. Respiratory bursts of prawn fed both forms of 1 mg Se (kg diet)(-1) significantly increased compared to control prawns. For the antioxidant status analysis, glutathione peroxidase, glutathione reductase, and glutathione s-transferase of prawn fed the SSe-enriched diet at 1 mg (kg diet)(-1) were significantly increased. The results indicated that the cheaper selenium, SSe is recommended to be added in prawn feed at the concentration of 0.5 mg resulting in 1.5 mg SSe (kg diet)(-1) increased prawn immunity and disease resistance against the pathogen, D. hansenii. PMID:20561587
Proteomic analysis of differentially expressed protein in hemocytes of wild giant freshwater prawn Macrobrachium rosenbergii infected with infectious hypodermal and hematopoietic necrosis virus (IHHNV)
Alinejad, T.; Bin, Kwong Q.; Vejayan, J.; Othman, R.Y.; Bhassu, S.
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
Proteomic analysis of differentially expressed protein in hemocytes of wild giant freshwater prawn Macrobrachium rosenbergii infected with infectious hypodermal and hematopoietic necrosis virus (IHHNV).
Alinejad, T; Bin, Kwong Q; Vejayan, J; Othman, R Y; Bhassu, S
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
Wen-Liang Chen; Hung-Hung Sung
A previous in vitro study has indicated that four phthalate esters (PAEs) could damage hemocytes and decreases the cellular immunity of prawns [Sung, H.H., Kao, W.Y., Su, Y.J., 2003. Effects and toxicity of phthalate esters to hemocytes of giant freshwater prawn, Macrobranchium rosenbergii. Aquat. Toxicol. 64, 25–37]. The aim of this study was to investigate the in vivo effect of
Vázquez-Acevedo, Nietzell; Reyes-Colón, Dalynés; Ruíz-Rodríguez, Eduardo A.; Rivera, Nilsa M.; Rosenthal, Joshua; Kohn, Andrea B.; Moroz, Leonid L.; Sosa, María A.
Biogenic amines are implicated in several mental disorders, many of which involve social interactions. Simple model systems, such as crustaceans, are often more amenable than vertebrates for studying mechanisms underlying behaviors. Although various cellular responses of biogenic amines have been characterized in crustaceans, the mechanisms linking these molecules to behavior remain largely unknown. Observed effects of serotonin receptor agonists and antagonists in abdomen posture, escape responses, and fighting have led to the suggestion that biogenic amine receptors may play a role in modulating interactive behaviors. As a first step in understanding this potential role of such receptors, we have cloned and fully sequenced two serotonin receptors, 5-HT1Mac and 5-HT2Mac, from the CNS of the freshwater prawn Macrobrachium rosenbergii, and have mapped their CNS immunohistochemical distribution. 5-HT1Mac was found primarily on the membranes of subsets of cells in all CNS ganglia, in fibers that traverse all CNS regions, and in the cytoplasm of a small number of cells in the brain, circum- and subesophageal ganglia (SEG), most of which also appear to contain dopamine. The pattern of 5-HT2Mac immunoreactivity was found to differ significantly, being found mostly in the central neuropil area of all ganglia, in glomeruli of the brain’s olfactory lobes, and in the cytoplasm of a small number of neurons in the SEG, thoracic and some abdominal ganglia. The observed differences in terms of localization, distribution within cells, and intensity of immunoreactive staining throughout the prawn’s CNS suggest that these receptors are likely to play different roles. PMID:19184976
Puthawibool, Teeranart; Senapin, Saengchan; Flegel, Timothy W; Kiatpathomchai, Wansika
Loop-mediated isothermal amplification (LAMP) allows rapid amplification of nucleic acids under isothermal conditions. It can be combined with a chromatographic lateral flow dipstick (LFD) for much more efficient, field-friendly detection of MrNV. In this work, RT-LAMP was performed at 65 degrees C for 40 min, followed by 5 min for hybridization with an FITC-labeled DNA probe and 5 min for LFD resulted in visualization of DNA amplicons trapped at the LFD test line. Thus, total assay time, including 10 min for rapid RNA extraction was approximately 60 min. In addition to advantages of short assay time, confirmation of amplicon identity by hybridization and elimination of electrophoresis with carcinogenic ethidium bromide, the RT-LAMP-LFD was more sensitive than an existing RT-PCR method for detection of MrNV. The RT-LAMP-LFD method gave negative test results with nucleic acid extracts from normal shrimp and from shrimp infected with other viruses including DNA viruses [PstDNV (IHHNV), PemoNPV (MBV), PmDNV (HPV), WSSV] and RNA viruses (TSV, IMNV, YHV/GAV). PMID:20655379
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.
Rattanavichai, Wutti; Cheng, Winton
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
, turbidity and other organic substances was very effective, but no significant reduction in ammonia nitrogen content wss achieved, unless an oyster shell filter was introduced in the media. fea lty f tf 1 * 9 11 9 b*Mt ~btt demonstrated. Ozone... in the concentration range of 7 to 14 ppb was found t b btfhly *t M. ~b* tt. Possible mechanisms for the observed reaction of ozone are discussed. iv ACKNOWLDEGEMENTS I thank Dr. Richard Weaver and Dr. Richard Noble for their guidance in this study. Dr. Robert...
the basic Lowther plate principle. The rate of ozone input into a solution by the generator was found to be 0. 62 mg/min under an applied power supply of 14 watts/hour. Ozonation of culture media was analyzed. Ozonation of COD including ni. trite... or at 450 ppm COD. 18 7. Effects of ozonation on NH3-N in culture radium made from dechlorinated tap water 20 8. Effects of ozonation on COD in culture medium made from dechlorinated tap water. 22 9. Effects of ozone on COD and NH3-N in culture medium...
Dai, Xilin; Xiong, Zhaodi; Xie, Jian; Ding, Fujiang
Endosulfan, an organochlorine pesticide, is highly toxic and effective at controlling pests in agriculture, horticulture, and public health programs. In this study, static bioassays were used to evaluate the toxicity of endosulfan to freshwater prawns ( Macrobrachium rosenbergii) of various lengths (1.5±0.03, 4±0.08, and 7±0.06 cm). Additionally, the activities of peroxidase (POD), acid phosphatase (ACP), alkaline phosphatase, acetylcholinesterase (AChE), and Na+/K+-ATPase were analyzed to reflect the effects of endosulfan exposure. The 96 h LC50 of endosulfan for prawns 1.5, 4, and 7 cm long were 1.86, 4.53, and 6.09 ?g/L, respectively, improved tolerance to endosulfan with growth. The POD activities of test organisms exposed to low concentrations of endosulfan were inhibited, indicating the presence of oxygen damaged tissue. Moreover, a notable decrease in AChE activity was observed due to overstimulation of neurotransmission, which might result in abnormal behavior. The effect caused by endosulfan on phosphatase production in the hepatopancreas of prawns 1.5, 4, and 7 cm long was different because the ability of nonspecific immune regulation increased with growth. The 96 h LC50 values obtained in this study could be used in the formulation of water-quality criteria in China. Moreover, the changes in enzymes activities of M. rosenbergii under stress of endosulfan could be applied in the establishment of early warning indicators for bio-safety.
Qian, Ye-Qing; Li, Ye; Yang, Fan; Yu, Yan-Qin; Yang, Jin-Shu; Yang, Wei-Jun
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
Starkey, William G; Millar, Rose Mary; Jenkins, Mary E; Ireland, Jacqueline H; Muir, K Fiona; Richards, Randolph H
Nucleic acid sequence based amplification (NASBA) is an isothermal nucleic acid amplification procedure based on target-specific primers and probes, and the co-ordinated activity of 3 enzymes: AMV reverse transcriptase, RNase H, and T7 RNA polymerase. We have developed a real-time NASBA procedure for detection of piscine nodaviruses, which have emerged as major pathogens of marine fish. Viral RNA was isolated by guanidine thiocyanate lysis followed by purification on silica particles. Primers were designed to target sequences in the nodavirus capsid protein gene, yielding an amplification product of 120 nucleotides. Amplification products were detected in real-time with a molecular beacon (FAM labelled/methyl-red quenched) that recognised an internal region of the target amplicon. Amplification and detection were performed at 41 degrees C for 90 min in a Corbett Research Rotorgene. Based on the detection of cell culture-derived nodavirus, and a synthetic RNA target, the real-time NASBA procedure was approximately 100-fold more sensitive than single-tube RT-PCR. When used to test a panel of 37 clinical samples (negative, n = 18; positive, n = 19), the real-time NASBA assay correctly identified all 18 negative and 19 positive samples. In comparison, the RT-PCR procedure identified all 18 negative samples, but only 16 of the positive samples. These results suggest that real-time NASBA may represent a sensitive and specific diagnostic procedure for piscine nodaviruses. PMID:15212274
Yeh, Shinn-Pyng; Liu, Kuan-Fu; Chiu, Shieh-Tsung; Jian, Shun-Ji; Cheng, Winton; Liu, Chun-Hung
A selenium dependent glutathione peroxidase (Se-GPx) cDNA was cloned from haemocyte by a reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA (RACE). The 913 bp cDNA contained an open reading frame (ORF) of 558 bp encoded a deduced amino acid sequence of 186 amino acids. The prawn Se-GPx sequence contains a selenocysteine (Sec) residue which is encoded by the unusual stop codon, (115)TGA(117). According to the molecular modeling analysis, the active site Sec residue, located in the loop between beta3 and alpha2 in a pocket on the protein surface, and hydrogen bonded to Gln(73) and Trp(141). A GPx signature motif 2, (63)LAFPCNQF(70) and active site motif, (151)WNFEKF(156), two arginine (R) residues, R(89) and R(167) contribute to the electrostatic architecture that directs the glutathione donor substrate, and two putative N-glycosylation site, (75)NNT(77) and (107)NGS(109) were observed in the prawn Se-GPx sequence. In addition, the eukaryotic selenocysteine insertion sequence element is conserved in the 3'-UTR. Comparison of amino acid sequences showed that prawn Se-GPx is more closely related to vertebrate GPx 1. The prawn Se-GPx was synthesized in haemocyte, hepatopancreas, muscle, stomach, gill, intestine, eyestalk, heart, epidermis, lymph organ, ventral nerve cord, testis and ovary. The increase of respiratory burst in haemocyte was observed in pathogen, Debaryomyces hansenii-injected prawn in order to kill the pathogen, and the up-regulation in SOD and GPx acitivity, and prawn Se-GPx mRNA transcription were involved with the protection against damage from oxidation. PMID:19376233
Llobrera, Jose Alvarez
concentrations of ammonia and other toxic substances as well as levels that can be safely maintained in the cul- ture system under a variety of environmental conditions. This will allow proper management of the system and efficient utilization of resources... not be the exclusive toxic agent. When IC50 values obtained for the three levels of pH (6. 83, 7. 6, and 8. 34) were expressed in terms of un-ionized ammonia in that study, it would appear that the larvae exposed to the lowest NH3 concentrations (pH 6. 83) were...
Newitt, Richard Allen
of labeled and unlabeled ecdysone through purification. Experiment 49: Ecdysteroid titer of prawn carcasses with determination of recovery. 38 39 TABLE 5. Experiment 410: HPLC analysis of zone E from TLC separations 45 TABLE 6. Experiment iI11... on scheme for ecdysteroi ds from car cass or ovaries Molt cycle stages A, 8, and C Molt cycle stages 0 and D I 0 Molt cycle stages Dl and Dl II III Molt cycle stage D3. 15 22 26 28 FIGURE 9: Contamination of HPLC system with ecdyster aids...
Llobrera, Alcestis Trillo
minutes exposure time 20 INTRODUCTION Fish populations maintained under crowded conditions inherent to aquaculture are subject to a variety of inf'ectious diseases (Hanson and Goodwin, 1977). Sometimes these diseases develop into epizootics causing.... and Amend, D. F. , 1978. Hyperosmotic infiltra- tiond Factors influencing uptake of bovine serum albumin by ' b * * )S 1 B d '). J. F' h. R* . B* d Can. , g5: 871-874. Goodwin, H. L. and Hanson, J. A. , 1975. The Aquaculture of Freshwater Prawns...
Newitt, Richard Allen
Whinnie et al. , 1972), Palaemonetes ~u io (Freeman and Bartell, 1975), 1976; Hubschman and Armstrong, 1972), Gammarus ~s . (Ducruet, 1975), Homarus americanus (Rao et al. , 1973), Procambrus ~s . (Lowe et al. , 1968; Krishnakumaran and Schneiderman, 1970...
Wang, Zhaowei; Qiu, Yang; Liu, Yongxiang; Qi, Nan; Si, Jie; Xia, Xiaoling; Wu, Di; Hu, Yuanyang; Zhou, Xi
Nodaviruses are a family of positive-stranded RNA viruses with a bipartite genome of RNAs. In nodaviruses, genomic RNA1 encodes protein A, which is recognized as an RNA-dependent RNA polymerase (RdRP) and functions as the sole viral replicase protein responsible for its RNA replication. Although nodaviral RNA replication has been studied in considerable detail, and nodaviruses are well recognized models for investigating viral RNA replication, the mechanism(s) governing the initiation of nodaviral RNA synthesis have not been determined. In this study, we characterized the RdRP activity of Wuhan nodavirus (WhNV) protein A in detail and determined that this nodaviral protein A initiates RNA synthesis via a de novo mechanism, and this RNA synthesis initiation could be independent of other viral or cellular factors. Moreover, we uncovered that WhNV protein A contains a terminal nucleotidyltransferase (TNTase) activity, which is the first time such an activity has been identified in nodaviruses. We subsequently found that the TNTase activity could function in vitro to repair the 3? initiation site, which may be digested by cellular exonucleases, to ensure the efficiency and accuracy of viral RNA synthesis initiation. Furthermore, we determined the cis-acting elements for RdRP or TNTase activity at the 3?-end of positive or negative strand RNA1. Taken together, our data establish the de novo synthesis initiation mechanism and the TNTase activity of WhNV protein A, and this work represents an important advance toward understanding the mechanism(s) of nodaviral RNA replication. PMID:24019510
Hsu, Hao-Hsuan; Lee, Szu-Hsien; Chen, Young-Mao; Tsai, Tieh-Jung; Ou, Ming-Chang; Ku, Hsiao-Tung; Lee, Gwo-Bin; Chen, Tzong-Yueh
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
Christopher, Frank Mitchell
For Foods (36 ) were followed. For the isolation and enumeration of coliform bacteria samples were introduced into Lauryl Sulfate Tryptone Broth (ISTB, BBL). Contents of positive tubes (gas formation within 48 hours at 35 C) were inoculated...
Kua, Beng Chu; Choong, F C; Hazreen Nita, M K; Muhd Faizul H, A H; Bhassu, S; Imelda, R R; Mohammed, M
A preliminary survey of parasitic and infectious hypodermal and haematopoietic necrosis virus (IHHNV) infections in giant freshwater prawn from the Damak Sea of Rejang River, Kuching, Sarawak was conducted. Symptoms of black spots/patches on the rostrum, carapace, pleopods or telson were observed in most of the 107 samples collected. Parasitic examination revealed sessiline peritrichs such as (Zoothamnium sp.), nematode larvae, gregarine stage and cocoon of leech with prevalences of 1.2%, 1.2%, 5% and 17% respectively. Under histopathological examination, changes like accumulation of hemocytes around hepatopancreatic tubules due to vibriosis, basophilic intranuclear inclusions in the epithelium and E-cell of hepatopancreatic tubules as a result of HPV were seen through the section. No positive infection of IHHNV was detected in 78 samples. As such, the wild giant freshwater prawns in Damak Sea of Rejang River in Kuching are IHHNV-free though infections of parvo-like virus and bacteria were seen in histopathology. PMID:21602773
Christopher, Frank Mitchell
to their initial level af- 2 ter 21-22 days and continued to increase with counts at 28 days ranging from 10 -10 per gram. Although Aeromonas 6 7 and Pseudomonas species continued to dominate the microbial flora after 7 days, Acinetobacter and Pseudomonas... the principal isolates from the drip. After 6 days storage, Moraxella-Acinetobacter, Micrococcus, and Cor ebacterium were the predominant genera. After 10 days, Pseudomonas dominated the microbial flora of the drip. These studies illustrate the relationship...
Geometric morphometrics of carapace of Macrobrachium australe (Crustacea: Palaemonidae) from Keywords: phenotypic plasticity, crustaceans, amphidr- omy, morphometrics, shape, carapace Accepted, V. 2012. Geometric morphometrics of carapace of Macrobrachium australe (Crustacea: Palaemonidae
Chen, Young-Mao; Kuo, Cham-En; Wang, Ting-Yu; Shie, Pei-Shiuan; Wang, Wei-Chen; Huang, Shao-Ling; Tsai, Tieh-Jung; Chen, Peng-Peng; Chen, Jiann-Chu; Chen, Tzong-Yueh
The heat shock proteins (HSPs) family which consists of HSP90, HSP70, and low molecular mass HSPs are involved in chaperone activity. Here, we report the cloning and characterization of HSP90AB gene from orange-spotted grouper, Epinephelus coioides. The full-length of grouper HSP90AB was 727 amino acids and possessed an ATPase domain as well as an evolutionarily conserved molecular chaperone. The HSP90AB-green fluorescent protein fusion protein was evenly distributed in the cytoplasm. Immunohistochemistry (IHC) and real-time polymerase chain reaction (PCR) analyses indicated that the expression of grouper HSP90AB was marginally increased following nodavirus infection. Grouper E. coioides that received HSP90 inhibitor geldanamycin (GA) showed an increase in HSP90AB expression and growth of nodavirus supporting nodavirus replication. PMID:20153436
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
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
Qin, Q W; Wu, T H; Jia, T L; Hegde, A; Zhang, R Q
The development and characterization of a new tropical marine fish cell line (GS), derived from the spleen of orange spotted grouper, Epinephelus coioides is described. The GS cells grow well in Leibovitz's L-15 medium supplemented with 10% foetal bovine serum, and have been subcultured more than 200 times. The optimal growth temperature was 27 degrees C. The GS cell culture consisted of mostly fibroblastic cells. The modal diploid chromosome number was 48. GS cell cultures showed advanced cytopathic effects after infection with a pathogenic grouper iridovirus (Singapore grouper iridovirus, SGIV) or with a grouper nodavirus (Epinephelus tauvina nervous necrosis virus, ETNNV). Analysis by transmission electron microscopy showed a large number of SGIV and ETNNV particles in the cytoplasm of virus-infected cells, respectively, indicative of high sensitivity to these two viruses. Immunofluorescence microscopy showed that iridovirus-infected GS cells reacted strongly with monoclonal antibody against the grouper iridovirus. It is suggested that the GS cell line has good potential as a diagnostic tool for isolation and propagation of iridovirus and nodavirus. When the GS cells were transfected with pEGFP vector DNA, significant fluorescent signals were observed suggesting that the GS cell line can be used as a useful tool for transgenic and genetic manipulation studies. PMID:16137774
Cai, Dawei; Qiu, Yang; Qi, Nan; Yan, Ran; Lin, Meijuan; Nie, Dongbo; Zhang, Jiamin; Hu, Yuanyang
Wuhan Nodavirus (WhNV) is the first reported nodavirus isolated from insect in China. The viral genome consists of two positive-strand RNA, RNA1 and RNA2. RNA1 is 3149 nucleotides in length, and contains three putative Open Reading Frames (ORFs) which encode proteins A, B1 and B2, respectively. In contrast, only one putative ORF encoding protein alpha was identified within 1562-nt-long RNA2 species. Here, we report the newly characterized molecular properties of WhNV subgenomic RNA3 and its encoded protein B2. We have successfully multiplied WhNV in the natural host Pieris rapae larvae under laboratory conditions. WhNV replication in the host cells resulted in the expression of viral proteins, ProA, B2 and Proalpha, with the absence of B1 production. Northern blot hybridization assay revealed the existence of subgenomic RNA3 which is 5' capped and 3' co-terminal with RNA1. The subgenomic RNA3 is 370 nucleotides in length and contains only one ORF (B2) with the first AUG as the authentic initiation codon. In addition, we found that nonstructural protein B2 of WhNV is an efficient RNA interference (RNAi) suppressor in a cultured drosophila cell line. The amino-terminal region (aa 1-20) of B2 is essential for this RNAi inhibition activity. PMID:20441781
Chen, Young-Mao; Kuo, Cham-En; Chen, Guan-Ru; Kao, Yu-Ting; Zou, Jun; Secombes, Chris J; Chen, Tzong-Yueh
We cloned and sequenced 2C I-IFN, a two-cysteine containing type I interferon (I-IFN) gene, in orange-spotted grouper (Epinephelus coioides). The cDNA has 769 base pairs, the protein has 172 amino acids, and the predicted signal peptide has 18 amino acids with two cysteines. This gene is similar to I-FNs from sea bass and other teleosts. 2C I-IFN has 5 exons and 4 introns, also similar to other teleost I-IFNs. Immunohistochemical (IHC) analysis indicated that expression is predominantly membrane-localized in healthy grouper, but has a zonal distribution in nodavirus-infected grouper. Grouper infected with nodavirus had elevated levels of 2C I-IFN at 72 h and Mx at days 6-7. Recombinant 2C I-IFN activated grouper Mx, leading to upregulated antiviral activity. The grouper Mx promoter was highly induced after treatment with recombinant 2C I-IFN. The present results suggest that expression of grouper 2C I-IFN may participate in the immunologic barrier function against nodavirus. PMID:24731841
Sokolow, Susanne H; Lafferty, Kevin D; Kuris, Armand M
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
P Saravana Bhavan; P Geraldine
The tissue damage induced by various organic pollutants in aquatic animals is well-documented, but there is a dearth of information relating to the histological alterations induced by pesticides in freshwater prawns of the genus Macrobrachium. In the present study intermoult juveniles of the freshwater prawn Macrobrachium malcolmsonii (total length, 4.5–5.0 cm; weight, 1.0–1.25 g) were exposed to three sublethal concentrations
Pinhong, Wang; Wei, Ji; Yaqing, Chang
An electrochemical technique for the real-time detection of hydrogen peroxide (H2O2) was employed to describe respiratory burst activity (RBA) of phagocytes in plasma which can be used to evaluate the ability of immune system and disease resistance. The method is based upon the electric current changes, by redox reaction on platinum electrode of extracellular hydrogen peroxide (H2O2) released from phagocytes stimulated by the zymosan at 680 mV direct current (d.c.). Compared with the control, activation of respiratory burst by zymosan particles results in a high amperometric response, and a current peak was obtained during the whole monitoring process. The peak current was proved by addition of Cu2+ and other controls, to be the result of intense release of H2O2 from phagocytes. The peak area was calculated and used to evaluate the quantity of effective H2O2, which represents the quantity of H2O2 beyond the clearance of related enzymes in plasma. According to Faraday's law, the phagocytes' ability of prawns to generate effective H2O2 was evaluated from 1.253 x 10(-14) mol/cell to 6.146 x 10(-14) mol/cell, and carp from 1.689 x 10(-15) mol/cell to 7.873 x 10(-15) mol/cell. This method is an acute and quick detection of extracellular effective H2O2 in plasma and reflects the capacity of phagocytes under natural conditions, which could be applied for selecting species and parents with high immunity for breeding in aquaculture. PMID:17728150
Qi, Nan; Cai, Dawei; Qiu, Yang; Xie, Jiazheng; Wang, Zhaowei; Si, Jie; Zhang, Jiamin; Zhou, Xi; Hu, Yuanyang
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 ?2 and ?3, while helix ?1 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
Nguyen Quoc An; Phan Dinh Phuc; Phan Thi Le Anh; Nguyen Thi Tu; Ly Ngoc Tuyen; Le Phuoc
Aquaculture of the freshwater prawn Macrobrachium nipponense has potential to develop into a high value enterprise. However, this commercial development faces the constraints of insufficient stocking material and a lack of suitable culture technology. This paper presents preliminary results of experiments designed to address these two problems. The experiments on prawn seed production used eight small (each 2 m2 in
S. Arun; P. Subramanian
Activities of superoxide dismutase, catalase, glutathione peroxidase (selenium-dependent and selenium-independent) and glutathione-S-transferases were analysed in freshwater prawn Macrobrachium malcolmsonii during embryonic and larval development. An elevated level of these enzymes was encountered in the larval stage of M. malcolmsonii when compared to its preceding embryonic stages. Enzyme activities were also analysed in hepatopancreas, muscle and gill of M. malcolmsonii sub-adults
P. Saravana Bhavan; P. Geraldine
This study was conducted to investigate the induction of biochemical stress responses in Macrobrachium malcolmsonii following exposure to endosulfan and to determine the most sensitive parameter of endosulfan-induced stress in this species of prawn. Intermolt juvenile prawns were exposed to three sublethal concentrations of endosulfan (10.6, 16.0, and 32.0 ng\\/L) for a period of 21 days. Samples were taken from
Nwosu, Francis M.
The Cross River Estuary, Nigeria, is an important shrimping area for artisanal fishermen of the coastal communities. The multi-species Macrobrachium fishery is exploited with three main gears, namely beach seine, push net and trap. Studies on species composition of this fishery recorded thirteen shrimp species, one swimming crab ( Callinectes amnicola) and two fish species ( Eleotris sp. and Pellonula leonensis). The shrimp species identified included Macrobrachium macrobrachion (83.39% and 55.69% by number and weight, respectively), M. vollenhovenii (9.66% and 37.18%), M. equidens (3.8% and 2.87%), juveniles-sub-adults of Penaeus notialis (1.11% and 1.3%), M. dux, M. felicinum, Palaemonetes africanus, Palaemon maculatus, Palaemon elegans, Desmocaris sp., Leander sp., Nematopalaemon hastatus and Alpheus pontederiae. While the selectivity index for trap was 0.25, beach seine and push net had a lower index of 0.063. The results present the first comprehensive and representative report for the Estuary shrimp fishery and will assist in the management of the biodiversity of this ecosystem.
Lal, Monal M; Seeto, Johnson; Pickering, Timothy D
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
Cresswell, Tom; Simpson, Stuart L; Smith, Ross E W; Nugegoda, Dayanthi; Mazumder, Debashish; Twining, John
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
In inquiry-based instruction, discovery and learning belong to the students. In this exploration, jumbo shrimp are the source of inspiration. The magic in this project lies not in successfully culturing these shrimp, known as Macrobrachium rosenbergii
Vacuolating encephalopathy and retinopathy associated with a nodavirus-like agent: a probable cause of mass mortality of wild Golden grey mullet (Liza aurata) and Sharpnose grey mullet (Liza saliens) in Iranian waters of the Caspian Sea.
Zorriehzahra, Mohammad Jalil; Nazari, Alireza; Ghasemi, Mohaddes; Ghiasi, Maryam; Karsidani, Somayeh Haghighi; Bovo, Giuseppe; Daud, Hassan Hj Mohd
Mullets are dominant fishes in the catch composition in the southern coasts of the Caspian Sea and after (Rutilus frisii kutum Kamensky, 1901) have a worthy role in production of marine proteins and incomings of north provinces of Iran. Mullets stocks decreased dramatically in recent decades in the Caspian Sea and catch amount reached from 6446 MT on 2002 to 2151 MT in 2012. Mysterious mortalities occurred in wild mullet (Liza auratu) and (Liza saliens) in Iranian waters of Caspian Sea in recent years. Regarding to investigation of causative agent of mentioned outbreak about 322 suspected samples were collected from coastal capture sites of Iranian north provinces in 2008 till 2011. Moribund fish revealed skin darkening, erratic swimming, belly-up at rest and high distension of swim bladder. Target tissues such as brain and eye were removed and then fixed for histopathology and TEM assay. Widespread and massive vacuolation were observed in brain, spinal cord, retina and optical nerve and intracytoplasmic vacuoles and virus particles in retina. So concerning to clinical signs, histopathological and TEM findings, it could be concluded that nodavirus-like agent could be probable cause of mass mortality of wild mullet in Iranian waters of the Caspian Sea. PMID:25674618
Lavarías, S; García, C; Crespo, R; Pedrini, N; Heras, H
Several agrochemicals like organophosphates are extensively used to control pests in agricultural practices but they also adversely affect non-target fauna. The effect of organophosphorous fenitrothion on the prawn Macrobrachium borellii was evaluated. The 96-h LC50 was determined. Activity of superoxide dismutase, catalase, glutathione-S-transferase and lipid oxidation levels, were evaluated in the hepatopancreas from adults exposed to sublethal fenitrothion concentrations for 1, 2, 4 and 7 days. In addition, superoxide dismutase mRNA expression, acetylcholinesterase inhibition and haemocyte DNA damage were determined. The 96-h LC50 was 4.24?g/l of fenitrothion. Prawn exposed to sublethal FS concentrations showed an increase of both catalase and superoxide dismutase activities, mainly after 2 and 4 days exposure and an increase of glutathione-S-transferase activity from day 2 to day 7 while lipid oxidation levels increased mainly on day 1. Superoxide dismutase transcripts were significantly higher in fenitrothion -treated prawns, indicating an induction mechanism. Hemolymph analysis showed that while acetylcholinesterase activity decreased after 2 days, haemocytes displayed most DNA damage after 7-day exposure to fenitrothion. These results indicate that prawn enzymes are highly sensitive to fenitrothion exposure, and these biological responses in M. borellii could be valuable biomarkers to monitor organophosphorous contamination in estuarine environments. PMID:23876938
Pasquevich, M Y; Dreon, M S; Gutierrez Rivera, J N; Vázquez Boucard, C; Heras, H
Hydrocarbon pollution is a major environmental threat to ecosystems in marine and freshwater environments, but its toxicological effect on aquatic organisms remains little studied. A proteomic approach was used to analyze the effect of a freshwater oil spill on the prawn Macrobrachium borellii. To this aim, proteins were extracted from midgut gland (hepatopancreas) of male and female prawns exposed 7 days to a sublethal concentration (0.6 ppm) of water-soluble fraction of crude oil (WSF). Exposure to WSF induced responses at the protein expression level. Two-dimensional gel electrophoresis (2-DE) revealed 10 protein spots that were differentially expressed by WSF exposure. Seven proteins were identified using MS/MS and de novo sequencing. Nm23 oncoprotein, arginine methyltransferase, fatty aldehyde dehydrogenase and glutathione S-transferase were down-regulated, whereas two glyceraldehyde-3-phosphate dehydrogenase isoforms and a lipocalin-like crustacyanin (CTC) were up-regulated after WSF exposure. CTC mRNA levels were further analyzed by quantitative real-time PCR showing an increased expression after WSF exposure. The proteins identified are involved in carbohydrate and amino acid metabolism, detoxification, transport of hydrophobic molecules and cellular homeostasis among others. These results provide evidence for better understanding the toxic mechanisms of hydrocarbons. Moreover, some of these differentially expressed proteins would be employed as potential novel biomarkers. PMID:23570752
Short-term responses of Oryzias latipes (Pisces: Adrianichthyidae) and Macrobrachium nipponense (Crustacea: Palaemonidae) to municipal and pharmaceutical waste water in Beijing, China: survival, behaviour, biochemical biomarkers
A. Gerhardt; L. Janssens de Bisthoven; Z. Mo; C. Wang; M. Yang; Z. Wang
Whole effluent toxicity was assessed for the fish Oryzias latipes and the prawn Macrobrachium nipponense for 18 h in a dilution series (0–66%) of the inflow and effluent of a municipal waste water treatment plant as well as waste water from a teramycin producing pharmaceutical industry, before, during and after a pilot laboratory purification process. The waste waters caused acute
S. Lavarías; H. Heras; R. J. Pollero
The effects of a water-soluble fraction of light crude oil dissolved in freshwater (WSF) on Macrobrachium borellii exposed at three life stages was evaluated. Adults, larvae (PL), and embryos were exposed to five levels of WSF for 96 h. At 48 and 72 h PL were significantly more sensitive to WSF than adults, though values for 96-h LC50 were not
Subcellular\\/tissue distribution and responses to oil exposure of the cytochrome P450-dependent monooxygenase system and glutathione S-transferase in freshwater prawns ( Macrobrachium malcolmsonii, M. lamarrei lamarrei )
S. Arun; A. Rajendran; P. Subramanian
Subcellular fractions (mitochondrial, cytosolic and microsomal) prepared from the tissues (hepatopancreas, muscle and gill) of freshwater prawns Macrobrachium malcolmsonii and Macrobrachium lamarrei lamarrei were scrutinized to investigate the presence of mixed function oxygenase (MFO) and conjugating enzymes (glutathione-S-transferase, GST). Cytochrome P450 (CYP) and other components (cytochrome b5; NADPH-cytochrome c (CYP) reductase and NADH-cytochrome c-reductase activities) of the MFO system were
Chen, Po-Cheng; Weng, Francis Cheng-Hsuan; Jean, Wen Dar; Wang, Daryi
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
S. Selvakumar; P. Geraldine; S. Shanju; T. Jayakumar
Agricultural run off that is contaminated with pesticides enters water bodies, thereby polluting the aquatic environment. The sensitivity of the freshwater prawn, Macrobrachium malcolmsonii, to such pesticides is well-documented. However, the stress response to sublethal concentrations of pesticides has scarcely been investigated. In the present study, the effect of two different sublethal concentrations (1\\/5th and 1\\/10th of LC50 value) of
P. Saravana Bhavan; P. Geraldine
Juveniles ofMacrobrachium malcolmsonii(4.5–5.0 cm in size and 1.0–1.25 g in weight) were exposed to three sublethal concentrations of endosulfan (10.6, 16.0, and 32.0 ng\\/L) for 21 days. Samples were taken from the hemolymph, hepatopancreas, gills, and muscle of representative prawns from each test and control group on the 1st, 8th, 15th, and 21st day after the start of the experiment.
Lavarías, S; Heras, H; Pedrini, N; Tournier, H; Ansaldo, M
The aim of the present work was to evaluate the effect of the water soluble fraction of hydrocarbons (WSF) on the antioxidant status of the freshwater prawn Macrobrachium borellii. First, seasonal variations were studied in a non-polluted area. Hepatopancreas and gills showed season-related fluctuations in catalase (CAT), glutathione-S-transferase (GST) activities and in lipid peroxidation levels (LPO), but not in superoxide dismutase (SOD). Then, adults were exposed semi-statically to sublethal doses for 7days. CAT, SOD, GST, and glutathione peroxidase (GPx) activities and LPO, reduced glutathione (GSH) and protein oxidation (PO) levels were determined. Exposed individuals showed significant increases in CAT, SOD, and GST activities in hepatopancreas and CAT activity in gills. GPx activity did not vary in either tissues. While LPO levels increased, GSH levels decreased significantly in hepatopancreas of exposed animals, but PO levels showed no variation. Induction of SOD was also assessed by Real-time PCR mRNA expression in hepatopancreas. The non-enzymatic antioxidant activity was also tested; ABTS 2,2'-azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid) was higher in hemolymph of treated-prawns compared to controls, but ferric reducing activity of plasma assay (FRAP) values did not change. Taken together, the present results indicated that the antioxidant defenses of M. borellii, mainly in hepatopancreas, were significantly affected by aquatic hydrocarbon contamination, regardless of the season. PMID:21320634
Zhang, Fengying; Chen, Liqiao; Qin, Jianguang; Zhao, Weihong; Wu, Ping; Yu, Na; Ma, Lingbo
The gustavus gene is required for localizing pole plasm and specifying germ cells. Research on gustavus gene expression will advance our understanding of the biological function of gustavus in animals. A cDNA encoding gustavus protein was identified and termed MnGus in the oriental river prawn Macrobrachium nipponense. Bioinformatic analyses showed that this gene encoded a protein of 262 amino acids and the protein belongs to the Spsb1 family. Real-time quantitative PCR analyses revealed that the expression level of MnGus in prawn embryos was slightly higher at the cleavage stage than at the blastula stage, and reached the maximum level during the zoea stage of embryos. The minimum level of MnGus expression occurred during the perinucleolus stage in the ovary, while the maximum was at the oil globule stage, and then the level of MnGus expression gradually decreased with the advancement of ovarian development. The expression level of MnGus in muscle was much higher than that in other tissues in mature prawn. The gustavus cDNA sequence was firstly cloned from the oriental river prawn and the pattern of gene expression was described during oocyte maturation, embryonic development, and in other tissues. The differential expression patterns of MnGus in the embryo, ovary and other somatic tissues suggest that the gustavus gene performs multiple physiological functions in the oriental river prawn. PMID:21359189
Qiao, H; Xiong, Y W; Jiang, S F; Fu, H T; Sun, S M; Jin, S B; Gong, Y S; Zhang, W Y
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
Rossi, Natália; Mantelatto, Fernando Luis
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
Zhang, W Y; Jiang, S F; Xiong, Y W; Fu, H T; Qiao, H; Sun, S M; Gong, Y S; Jin, S B
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
Juarez, L.M.; Sanchez, J. (Monterrey Institute of Technology, Sonora (Mexico))
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.
Nguyen Thanh Phuong; Tran Ngoc Hai; Tran Thi Thanh Hien; Tran Van Bui; Do Thi Thanh Huong; Vu Nam Son; Yoshinori Morooka; Yutaka Fukuda; Marcy N. Wilder
In Vietnam, the giant freshwater prawn Macrobrachium rosenbergii is becoming an increasingly important targeted species, as its culture, especially in rice fields, is considered to have\\u000a the potential to raise income among improverished farmers. The production of M. rosenbergii based on aquaculture reached over 10 000 tons per year in 2002, having increased from about 2500 tons since the 1990s.
Crispin, Taryn S; White, Craig R
Changes to an animal's abiotic environment-and consequent changes in the allometry of metabolic rate in the whole animal and its constituent parts-has considerable potential to reveal important patterns in both intraspecific and interindividual variation of metabolic rates. This study demonstrates that, after 6 wk of thermal acclimation at replicate treatments of 16°, 21°, and 25°C, standard metabolic rate (SMR) scales allometrically in Leichhardtian river prawns Macrobrachium tolmerum ([Formula: see text]) and that the scaling exponent and normalization constant of the relationship between SMR and body mass is not significantly different among acclimation treatments when measured at 21°C. There is, however, significant variation among individuals in whole-animal metabolic rate. We hypothesized that these observations may arise because of changes in the metabolic rate and allometry of metabolic rate or mass of organ tissues within the animal. To investigate this hypothesis, rates of oxygen consumption in a range of tissues (gills, gonads, hepatopancreas, chelae muscle, tail muscle) were measured at 21°C and related to the body mass (M) and whole-animal SMR of individual prawns. We demonstrate that thermal acclimation had no effect on organ and tissue mass, that most organ and tissue (gills, gonads, hepatopancreas) respiration rates do not change with acclimation temperature, and that residual variation in the allometry of M. tolmerum SMR is not explained by differences in organ and tissue mass and respiration rates. These results suggest that body size and ambient temperature may independently affect metabolic rate in this species. Both chelae and tail muscle, however, exhibited a reduction in respiration rate in animals acclimated to 25° relative to those acclimated to 16° and 21°C. This reduction in respiration rates of muscle at higher temperatures is evidence of a tissue-specific acclimation response that was not detectable at the whole-animal level. PMID:23799841
Xiu, Yunji; Wang, Yinghui; Jing, Yunting; Qi, Yakun; Ding, Zhengfeng; Meng, Qingguo; Wang, Wen
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
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
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
Hadi Zokaei Far; Roos B. Saad; Hassan Mohd Daud; Sharr Azni Harmin
The effect of Bacillus subtilis, isolated from digestive tract of Macrobrachium rosenbergii was investigated on growth and survival rate of Litopenaeus vannamei during 60 days of culture. Sixteen aquaria with four replicates were used for treatments and controls. Treatment groups were consisted of ?) shrimp fed diet with B. subtilis (T1), and ii) shrimp fed diet mixed with B. subtilis
Porntrai, Supaporn; Damrongphol, Praneet
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…
M. Wes Schilling; Juan L. Silva; Alessandra J. Pham; Taejo Kim; Louis R. DAbramo; Viodelda Jackson
Freshwater prawns, Macrobrachium rosenbergii, were harvested and transferred to holding tanks containing aerated well water. One group of prawns was held in the well water without any addition of salt (control). For post harvest salt acclimation, experimental treatments consisted of different salt sources (solar, halite, marine) that were initially added to achieve a salinity of 10 ppt. Then 5 ppt
Jin, Shubo; Fu, Hongtuo; Zhou, Qiao; Sun, Shengming; Jiang, Sufei; Xiong, Yiwei; Gong, Yongsheng; Qiao, Hui; Zhang, Wenyi
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
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.
Bai, Hongkun; Qiao, Hui; Li, Fajun; Fu, Hongtuo; Sun, Shengming; Zhang, Wenyi; Jin, Shubo; Gong, Yongsheng; Jiang, Sufei; Xiong, Yiwei
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
Immunocytochemical localization of V-H(+) -ATPase, Na(+) /K(+) -ATPase, and carbonic anhydrase in gill lamellae of adult freshwater euryhaline shrimp Macrobrachium acanthurus (Decapoda, Palaemonidae).
Maraschi, Anieli Cristina; Freire, Carolina Arruda; Prodocimo, Viviane
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. J. Exp. Zool. 323A: 414-421, 2015. © 2015 Wiley Periodicals, Inc. PMID:26036663
Hayes, A.; Johnson, W.C.
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)
Paisarn Sithigorngul; Nanthika Panchan; Parin Chaivisuthangkura; Siwaporn Longyant; Weerawan Sithigorngul; Amorn Petsom
Mouse antiserum against C-terminal amide of Pem-CMG (a peptide in the family of CHH\\/MIH\\/GIH) penta-deca peptide (RPRQRNQYRAALQRLamide=CMG-15) was generated and used for localization of the peptide in tissue and extract of the eyestalk of Penaeus monodon by means of immunohistochemistry and dot-ELISA in comparison with anti-T+ antiserum (T+=YANAVQTVamide: the putative C-terminal amide of crustacean hyperglycemic hormone (CHH) of Macrobrachium rosenbergii).
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.
Nguyen Thanh Phuong; Vu Nam; Vo Thanh Toan; Tran Thi; Thanh Hien; Pham Minh Duc
Trials of freshwater prawn (Macrobrachium rosenbergii) farming in rice field and garden ditch is being conducted at JIRCAS project research site, Tan Phu Thanh village, Chau Thanh A, Can Tho province. It includes three rice-prawn farms and one garden ditch prawn farm. Juvenile prawns of 0,045 g\\/prawn in average were stocked at a density of 2 prawn\\/m2 in rice-prawn farms,
, Tony Be´licard1 , Yanfang Jiang3 , Guoyan Zhao3 , Carl J. Franz3 , Leonard D. Goldstein2 , Mabel and C. briggsae with unusual morphological phenotypes in intestinal cells, we identified two novel RNA
Arranz, Silvia E; Avarre, Jean-Christophe; Balasundaram, Chellam; Bouza, Carmen; Calcaterra, Nora B; Cezilly, Frank; Chen, Shi-long; Cipriani, Guido; Cruz, V P; D'Esposito, D; Daniel, Carla; Dejean, Alain; Dharaneedharan, Subramanian; Díaz, Juan; Du, Man; Durand, Jean-Dominique; Dziadek, Jaros?aw; Foresti, F; Peng-cheng, Fu; Gao, Qing-bo; García, Graciela; Gauffre-Autelin, Pauline; Giovino, Antonio; Goswami, Mukunda; Guarino, Carmine; Guerra-Varela, Jorge; Gutiérrez, Verónica; Harris, D J; Heo, Moon-Soo; Khan, Gulzar; Kim, Mija; Lakra, Wazir S; Lauth, Jérémie; Leclercq, Pierre; Lee, Jeonghwa; Lee, Seung-Ho; Lee, Soohyung; Lee, Theresa; Li, Yin-hu; Liu, Hongbo; Liu, Shufang; Malé, Pierre-Jean G; Mandhan, Rishi Pal; Martinez, Paulino; Mayer, Veronika E; Mendel, Jan; Mendes, N J; Mendonça, F F; Minias, Alina; Minias, Piotr; Oh, Kyeong-Suk; Oliveira, C; Orivel, Jérôme; Orsini, L; Pardo, Belén G; Perera, A; Procaccini, G; Rato, C; Ríos, Néstor; Scibetta, Silvia; Sharma, Bhagwati S; Sierens, Tim; Singh, Akhilesh; Terer, Taita; Triest, Ludwig; Urbánková, So?a; Vera, Manuel; Villanova, Gabriela V; Voglmayr, Hermann; Vysko?ilová, Martina; Wang, Hongying; Wang, Jiu-li; Wattier, Rémi A; Xing, Rui; Yadav, Kamalendra; Yin, Guibo; Yuan, Yanjiao; Yun, Jong-Chul; Zhang, Fa-qi; Zhang, Jing-hua; Zhuang, Zhimeng
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
Pande, Gde Sasmita Julyantoro; Baruah, Kartik; Bossier, Peter
There currently is more and more interest in the use of natural products, such as tea polyphenols, as therapeutic agents. The polyphenol compound pyrogallol has been reported before to inhibit quorum-sensing-regulated bioluminescence in Vibrio harveyi. Here, we report that the addition of 10 mg · liter?1 pyrogallol protects both brine shrimp (Artemia franciscana) and giant river prawn (Macrobrachium rosenbergii) larvae from pathogenic Vibrio harveyi, whereas the compound showed relatively low toxicity (therapeutic index of 10). We further demonstrate that the apparent quorum-sensing-disrupting activity is a side effect of the peroxide-producing activity of this compound rather than true quorum-sensing inhibition. Our results emphasize that verification of minor toxic effects by using sensitive methods and the use of appropriate controls are essential when characterizing compounds as being able to disrupt quorum sensing. PMID:23545532
Tan, Tian Tian; Chen, Maoshan; Harikrishna, Jennifer Ann; Khairuddin, Norliana; Mohd Shamsudin, Maizatul Izzah; Zhang, Guojie; Bhassu, Subha
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
Ern, Rasmus; Huong, Do Thi Thanh; Phuong, Nguyen Thanh; Wang, Tobias; Bayley, Mark
In aquatic environments, rising water temperatures reduce water oxygen content while increasing oxygen demand, leading several authors to propose cardiorespiratory oxygen transport capacity as the main determinant of aquatic animal fitness. It has also been argued that tropical species, compared with temperate species, live very close to their upper thermal limit and hence are vulnerable to even small elevations in temperature. Little, however, is known about physiological responses to high temperatures in tropical species. Here we report that the tropical giant freshwater shrimp (Macrobrachium rosenbergii) maintains normal growth when challenged by a temperature rise of 6°C above the present day average (from 27°C to 33°C). Further, by measuring heart rate, gill ventilation rate, resting and maximum oxygen uptake, and hemolymph lactate, we show that oxygen transport capacity is maintained up to the critical maximum temperature around 41°C. In M. rosenbergii heart rate and gill ventilation rate increases exponentially until immediately below critical temperatures and at 38°C animals still retained more than 76% of aerobic scope measured at 30°C, and there was no indication of anaerobic metabolism at the high temperatures. Our study shows that the oxygen transport capacity is maintained at high temperatures, and that other mechanisms, such as protein dysfunction, are responsible for the loss of ecological performance at elevated temperatures. PMID:24198257
Mohd-Shamsudin, Maizatul Izzah; Kang, Yi; Lili, Zhao; Tan, Tian Tian; Kwong, Qi Bin; Liu, Hang; Zhang, Guojie; Othman, Rofina Yasmin; Bhassu, Subha
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
Arockiaraj, Jesu; Kumaresan, Venkatesh; Bhatt, Prasanth; Palanisamy, Rajesh; Gnanam, Annie J; Pasupuleti, Mukesh; Kasi, Marimuthu; Chaurasia, Mukesh Kumar
In this study, we reported a complete molecular characterization including bioinformatics features, gene expression, peptide synthesis and its antimicrobial activities of an anti-lipopolysaccharide (LPS) factor (ALF) cDNA identified from the established cDNA library of freshwater prawn Macrobrachium rosenbergii (named as MrALF). The mature protein has an estimated molecular weight of 11.240 kDa with an isoelectric point of 9.46. The bioinformatics analysis showed that the MrALF contains an antimicrobial peptide (AMP) region between T54 and P77 with two conserved cysteine residues (Cys55 and Cys76) which have an anti-parallel ?-sheet confirmation. The ?-sheet is predicted as cationic with hydrophobic nature containing a net charge of +5. The depicted AMP region is determined to be amphipathic with a predicted hydrophobic face 'FPVFI'. A highest MrALF gene expression was observed in hemocytes and is up-regulated with virus [white spot syndrome baculovirus (WSBV)], bacteria (Aeromonas hydrophila) and Escherichia coli LPS at various time points. The LPS binding region of MrALF peptide was synthesized to study the antimicrobial property, bactericidal efficiency and hemolytic capacity. The peptide showed antimicrobial activity against both the Gram-negative and Gram-positive bacteria. The bactericidal assay showed that the peptide recognized the LPS of bacterial cell walls and binding on its substrate and thereby efficiently distinguishing the pathogens. The hemolytic activity of MrALF peptide is functioning in a concentration dependant manner. In summary, the comprehensive analysis of MrALF showed it to be an effective antimicrobial peptide and thus it plays a crucial role in the defense mechanism of M. rosenbergii. PMID:24269604
To determine the distribution of two systems of centrifugal neurons innervating the soma clusters of the olfactory midbrain across decapod crustaceans, brains of the following nine species comprising most infraorders were immunostained with antibodies against dopamine and the neuropeptides substance P and FMRFamide: Macrobrachium rosenbergii, Homarus americanus, Cherax destructor, Orconectes limosus, Procambarus clarkii, Astacus leptodactylus, Carcinus maenas, Eriocheir sinensis and Pagurus bernhardus. One system consisting of several neurons with dopamine-like immunoreactivity that originate in the eyestalk ganglia was present in the four crayfish but not in any other species. These neurons project mainly into the lateral soma clusters (cluster 10) comprising the somata of ascending olfactory projection neurons and innervate very sparsely the medial soma clusters (clusters 9 and 11) containing the somata of local interneurons. In the innervation pattern of the lateral cluster, the dopamine-immunoreactive neurons showed large species-specific differences. The other system comprises a pair of giant neurons with substance P-like immunoreactivity. These neurons have somata in the median protocerebrum of the central brain and major projections into the lateral clusters and the core of the olfactory lobes, the neuropils that are the first synaptic relay in the central olfactory pathway of decapods; minor arborizations are present in the medial clusters. The system of substance P-immunoreactive giant neurons was present and of great morphological similarity in all studied species. Only in one species, the shrimp Macrobrachium rosenbergii, evidence for co-localization of FMRFamide-like with substance P-like immunoreactivity in these neurons was obtained. These and previously collected data indicate that the centrifugal neurons with dopamine-like immunoreactivity may be associated with the presence of an accessory lobe, a second-order neuropil that receives input from the olfactory lobe and only occurs in spiny lobsters, clawed lobsters and crayfish. The pair of centrifugal giant neurons with substance P-like immunoreactivity, on the other hand, appears to be a constitutive component of the decapod crustacean brain that most likely is functionally associated with the olfactory lobe. Both systems apparently exert modulatory functions on olfactory information processing by preferentially targeting the somata of the projection neurons. Thus, in the olfactory projection neurons, the somata seem to be more directly involved in information processing than in most other neurons of the arthropod CNS. PMID:9106436
Jarapala, Sreenivasa Rao; Kandlakunta, Bhaskarachary; Thingnganing, Longvah
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
S Arun; P Krishnamoorthy; P Subramanian
Oxidative stress stimulates production of hydrogen peroxide and organic hydroperoxides which are involved in lipid peroxidation and oxidative damage to protein. Glutathione peroxidase protects against oxidative stress. Quantitative data on glutathione peroxidase of crustaceans are very limited and so this study was performed to determine the assay condition of glutathione peroxidase in crustaceans. Particularly, some properties of hepatic selenium dependent
Yamuna, A; Bhavan, P Saravana; Geraldine, P
Healthy juveniles of M. malcolmsoniiwere exposed to 24.1 microg l(-1) of mercury (96 hr LC50: 145 microg l(-1) Hg) for a period of 21 days. The hepatopancreas and gills of the prawns were sampled on 8th, 15th and 22nd day of exposure. Accumulation and elimination of Hg, activity of glutathione S-transferase (GST), content of glutathione (GSH) and metallothionein (MT) level were studied. Mercury accumulation was found to be higher in the hepatopancreas (88.60 microg g(-1)) and lower in the gills (67.8 microg g(-1)). However, Hg elimination was found to be faster in the gills (62%) and slower in the hepatopancreas (58%). Therefore, the rate of Hg elimination did not match the rate of its uptake. The activity of GST was found to be higher in tissues of test prawns (5.94-9.13 nmol mg(-1) protein min(-1)) on all sampling days when compared with controls (3.454.23 nmol mg(-1) protein min(-1)). Similarly, the content of GSH was found to be higher in tissues of test prawns (0.80-1.43 micromol g(-1) protein) on all sampling days when compared with controls (0.55-1.00 micromol g(-1) protein). These results indicate the formation of glutathione conjugate in test prawns to eliminate Hg. The induction of MT level was also found to be higher in tissues of test prawns (57.50-75.76 nmol g(-1) protein) on all sampling days when compared with control (20.24-45.22 nmol g(-1) protein). This indicates the fact that sequestration of Hg has occurred for its easy elimination. Thus, induction of GST-GSH and MT ensured protection and adaptation of test prawns to thrive in Hg contaminated environment. PMID:23033656
will fulfill the requirements of the animal. Therefore, in order to minimize wastage of food and thus reduce the cost of feeding, each feed ingredient must be evaluated in terms of the ability of the animal to digest and absorb it. Knowledge of availability... and can decay, adversely affecting water quality. Bacterial growth on unconsumed food can further stress the culture system (Shlesser and Gallagher, 1974). In order to obtain maximum growth of prawns, feed should not only be of high nutritional quality...
Miyajima, Lester Shigemi
osmotically active constituents In crustacean hemolymph, free amino acids constitute 50%. The level of osmotic regulation varies with dif- ferent organisms. For example, a euryhaline species such as Eriocheir sinensis actively regulates the internal... and Florkln (1961). There is also some similarity in free amino acid content to the euryhaline crab, Wiocheir sinensis in freshwater (Brieteaux-Gregorle et al, 1962). The few studies with crayfish usually fail to include such pertinent data as ionic...
: Dr. Robert R. Stickney The effects of feeding rate and feeding frequency on protein and total organic matter absorption as well as dry matter ingestion h I h*t 9 Il *b h' ~9*dt, * 1 t'9 d. Feeding rate and feeding frequency did not appear to have... appreciation is expressed to Dr. Robert R. Stickney, chairman of my advisory committee, for his patience, guidance, encouragement and suppozt throughout the course of these studies and in the preparation of the manuscript. I am grateful to Dr. Darrell A...
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
Liu, Kuan-Fu; Yeh, Maw-Sheng; Kou, Guang-Hsiung; Cheng, Winton; Lo, Chu-Fang
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
Thai, Truong Quoc; Wille, Mathieu; Garcia-Gonzalez, Linsey; Sorgeloos, Patrick; Bossier, Peter; De Schryver, Peter
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 1,000 mg L(-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 mg L(-1) PHB powder suspension, while Artemia enriched in 10 mg L(-1) A80, 100 mg L(-1) A10, and 1,000 mg L(-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
Jezzini, Sami H; Reyes-Colón, Dalynés; Sosa, María A
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
Li, Jitao; Chen, Ping; Liu, Ping; Gao, Baoquan; Wang, Qingyin; Li, Jian
An extracellular CuZnSOD cDNA was cloned from the haemocytes of swimming crab Portunus trituberculatus by a reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA end (RACE) method. Analysis of the nucleotide sequence revealed that the ecCuZnSOD full-length cDNA consisted of 965 bp with an open reading frame of 579 bp. It encoded a polypeptide of 192 amino acids which had a predicted molecular weight of 20.0 kDa and with an estimated pI of 6.23. The deduced amino acid sequence contained a putative signal peptide of 31 amino acids. It is predicted to possess all the expected features of CuZnSOD members, including amino acids responsible for binding Cu and Zn, two putative CuZnSOD signatures, two cysteines and one N-linked glycosylation site. Sequence comparison showed that the CuZnSOD deduced amino acid sequence of P. trituberculatus has similarity of 80%, 76%, 55% and 50% to that of blue crab Callinectes sapidus, mud crab Scylla serrata, crayfish Pacifastacus leniusculus and freshwater prawn Macrobrachium rosenbergii, respectively. The ecCuZnSOD transcripts expressed in all examined tissues, including haemocytes, hepatopancreas, heart, stomach, intestine, gill, ovary and muscle. RT-PCR analysis indicated that ecCuZnSOD transcripts both in haemocytes and hepatopancreas increased in the first 3 h after Vibrio alginolyticus challenging, as the bacterial infection progressed, the challenged crabs decreased to levels significantly lower than control by 96 h post-infection. These facts indicated that ecCuZnSOD was potentially involved in the acute response against invading bacteria in P. trituberculatus. PMID:20848212
Ko, Chi-Fong; Chiou, Tzu-Ting; Vaseeharan, Baskaralingam; Lu, Jenn-Kan; Chen, Jiann-Chu
A prophenoloxidase (proPO) cDNA was cloned from the haemocytes of mud crab Scylla serrata using oligonucleotide primers and RT-PCR. Both 3'- and 5'-regions were isolated by rapid amplification of cDNA end (RACE) method. Analysis of the nucleotide sequence revealed that the cDNA clone has a full length of 2663bp, with an open reading frame of 2019bp, a 124-bp 5'-untranslated region, and a 520-bp 3'-untranslated region containing a poly A signal. It encodes a protein of 673 amino acids with a predicted molecular weight of 77.5kDa and with an estimated pI of 5.96. It contains two putative tyrosinase copper-binding motifs with six histidine residues (copper A, 185, 189, 211, and copper B, 346, 350, 386). The proPO has thiol-ester-like motif (GCGWPQHM), which showed similar structural features of proPOs from other decapod crustaceans. It also contains five possible glycosylation sites, and a conserved C-terminal region common to all known proPOs. Sequence comparison showed that the proPO-deduced amino acid of mud crab S. serrata has an overall similarity of 78%, 57%, 56%, 51-55%, 54%, 53%, 52%, 52%, and 52% to that of Dungeness crab Cancer magister, American lobster Homarus americanus, European lobster Homarus gammarus, kuruma prawn Marsupenaeus japonicus, crayfish Pacifastacus leniusculus, white shrimp Litopenaeus vannamei, tiger shrimp Penaeus monodon, green tiger shrimp Penaeus semisulcatus, and giant freshwater prawn Macrobrachium rosenbergii, respectively. The proPO was strongly expressed in haemocytes, but not in heart, eyestalk, gill, muscle, ovary, hepatopancreas, stomach, and intestine. The proPO transcript of mud crab S. serrata increased significantly in 12 and 24h post-lipopolysaccharide (LPS) injection, but returned to the original values in 72h post injection. PMID:16806468
Chaurasia, Mukesh Kumar; Palanisamy, Rajesh; Harikrishnan, Ramasamy; Arasu, Mariadhas Valan; Al-Dhabi, Naif Abdullah; Arockiaraj, Jesu
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 ?-helices (52.2%) which are connected over random coils (47.7%). The 3D structure of MrBOK showed three central helices (?6, ?7 and ?8) which formed the core of the protein and are flanked on one side by ?1, ?2 and ?3, and on the other side by ?4, ?5 and ?11. 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
Rajendran, K V; Makesh, M; Karunasagar, I
Among the viruses infecting penaeid shrimp, monodon-type baculovirus (MBV) otherwise known as Penaeus monodon singly enveloped nuclear polyhedrosis virus (PmSNPV), is one of the widely reported and well described viruses. It is a rod-shaped, enveloped, double-stranded DNA virus, and considered till recently, as the type A baculovirus. Besides MBV, two strains of SNPV are reported-plebejus baculovirus and bennettae baculovirus. MBV was reported to be originated from Taiwan and has wide geographic distribution and is reported to be enzootic in wild penaeids of the Indo-pacific coasts of Asia. The virus also has diverse host-range including a variety of cultured and captured shrimp species and freshwater prawn, Macrobrachium rosenbergii. MBV has been reported in all life stages of P. monodon with late larval, postlarval and young juvenile as the most susceptible stages/ages. However, MBV has not been documented in early larval stages. Although MBV has been reported to be tolerated well by shrimp, the infection has been attributed to decreased productivity. The target organs or tissues of MBV are the hepatopancreatic tubules and duct epithelium of postlarvae, juveniles and adults, and the anterior midgut epithelium of very young postlarvae. The prominent clinical sign of infection is the presence of multiple spherical inclusion bodies in the hepatopancreas and midgut epithelial cells. The major mode of transmission of the virus is horizontal through oral exposure to occlusion bodies, contaminated tissues or fomites. Minor morphometric variation of the virus has been reported among different isolates. The rod-shaped enveloped virus particles range from 265-324 nm in length and 42-77 nm in diameter. Although complete genome sequence of MBV is not available, nucleic acid of MBV is circular, double-stranded DNA with a genome size ranging from 80 to 160 kbp. The virus codes for a 53 kDa major polyhedrin polypeptide and two minor 47 and 49 kDa polypeptides. A variety of diagnostic tools have been reported for this virus including real-time PCR and LAMP-based detection. Taxonomic position is still uncertain and International Committee on Taxonomy of Viruses lists MBV as a tentative species named PemoNPV in the genus Nucleopolyhedrovirus. However, according to the latest genomic information on the virus, it has been suggested to create a new group of non-occluded bacilliform viruses called nudiviruses with MBV as one of the members. The aim of the current work is to describe the knowledge on the status, distribution and host-range, pathology, transmission, virus structure and morphogenesis, genomic characteristics, diagnosis and the latest taxonomic position of MBV. PMID:23997439
Leone, Francisco A.; Bezerra, Thais M. S.; Garçon, Daniela P.; Lucena, Malson N.; Pinto, Marcelo R.; Fontes, Carlos F. L.; McNamara, John C.
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 ?-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
Jiang, Hongbing; Franz, Carl J.; Wu, Guang; Renshaw, Hilary; Zhao, Guoyan; Firth, Andrew E.; Wang, David
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. PMID:24503084
Johnson, Karyn N.; Tang, Liang; Johnson, John E.; Ball, L. Andrew
The genome of some icosahedral RNA viruses plays an essential role in capsid assembly and structure. In T=3 particles of the nodavirus Pariacoto virus (PaV), a remarkable 35% of the single-stranded RNA genome is icosahedrally ordered. This ordered...
Toubanaki, Dimitra K; Margaroni, Maritsa; Karagouni, Evdokia
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
DeRisi, Joseph L.
We report here the draft genome sequence of tombunodavirus UC1 assembled from metagenomic sequencing of organisms in San Francisco wastewater. This virus shares hallmarks of members of the Tombusviridae and the nodavirus-like Plasmopara halstedii and Sclerophthora macrospora viruses. PMID:26139709
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...
Adrienne Burns; Keith F. Walker
Stable water levels and turbidity associated with flow regulation in the River Murray have promoted the growth of filamentous green algae and Cyanobacteria in biofilms on submerged wood. We investigated the assimilation of biofilm algae by two dominant consumers, the decapod crustaceans Macrobrachium australiense (Palaemonidae) and Paratya australiensis (Atyidae), in two river reaches differing in the extent of floodplain development,
Karen H. Watanabe; Frank W. Desimone; Arunthavarani Thiyagarajah; William R. Hartley; Albert E. Hindrichs
Between 1990 and 1994, samples of three shellfish species (i.e. blue crab, Callinectes sapidus;crayfish, Procambarus acutis; and river shrimp, Macrobrachium ohionii) and 16 fish species and were collected at six sites along the lower Mississippi River by the Louisiana Department of Environmental Quality, Office of Water Resources in coordination with the US Environmental Protection Agency. The fish species included: bigmouth
A. V. Yakovleva; V. A. Yakovlev
On the basis of data for the period 1998–2008, a faunistic review of invasive species of zoobenthos in the upper reaches of\\u000a the Kuybyshev Reservoir is given. In total, 30 species are revealed: polychaetes—3; oligochaetes—2; leeches—1; mollusks—5;\\u000a crustaceans—19 (gammarids—9; corophiums—2; cumaceans and mysids—3 each; narrow-clawed crayfish Astacus leptodactilus and shrimp Macrobrachium nipponense). Invasive species account for 10.8% of the total
Moulton, T.; de Souza, M. L.; Brito, E. F.; Krsulovic, F. M.; Silveira, R. M.; Ometto, J. B.
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 ? 13C corresponding to microalgae from the periphyton). Baetids appeared to be algal herbivores and both Potimirim and Macrobrachium appeared (from their ? 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.
Zeddam; Rodriguez; Ravallec; Lagnaoui
A small isometric virus has been isolated from larvae of the sweetpotato pest Spodoptera eridania (Cramer) collected near Pariacoto, Ancash province, Peru. It is designated the Pariacoto virus (PaV). In addition to its high pathogenicity on its natural host Spodoptera eridania, PaV was found to replicate in Spodoptera ochrea (Hampson) larvae but not in Spodoptera frugiperda (Smith) larvae. The size of the viral particle was estimated to be about 30 nm in diameter. Polyacrylamide gel electrophoresis showed a protein of approximately 40.5 kDa. After agarose gel electrophoresis, the viral genome appeared to be bipartite RNA. Gel immunodiffusion tests showed no serological relationship between PaV and Nodamura virus, the type species for insect nodaviruses. Electron microscopy confirmed that viral replication occurs in the cytoplasm. These properties are similar to those of other members of family Nodaviridae, to which the virus is currently assigned. Copyright 1999 Academic Press. PMID:10534414
Marsh, Elizabeth K.
The nematode Caenorhabditis elegans has been a powerful experimental organism for almost half a century. Over the past 10 years, researchers have begun to exploit the power of C. elegans to investigate the biology of a number of human pathogens. This work has uncovered mechanisms of host immunity and pathogen virulence that are analogous to those involved during pathogenesis in humans or other animal hosts, as well as novel immunity mechanisms which appear to be unique to the worm. More recently, these investigations have uncovered details of the natural pathogens of C. elegans, including the description of a novel intracellular microsporidian parasite as well as new nodaviruses, the first identification of viral infections of this nematode. In this review, we consider the application of C. elegans to human infectious disease research, as well as consider the nematode response to these natural pathogens. PMID:22286994
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
Wu, Wenzhe; Wang, Zhaowei; Xia, Hongjie; Liu, Yongxiang; Qiu, Yang; Liu, Yujie; Hu, Yuanyang; Zhou, Xi
Flock House virus (FHV) is a positive-stranded RNA virus with a bipartite genome of RNAs, RNA1 and RNA2, and belongs to the family Nodaviridae. As the most extensively studied nodavirus, FHV has become a well-recognized model for studying various aspects of RNA virology, particularly viral RNA replication and antiviral innate immunity. FHV RNA1 encodes protein A, which is an RNA-dependent RNA polymerase (RdRP) and functions as the sole viral replicase protein responsible for RNA replication. Although the RNA replication of FHV has been studied in considerable detail, the mechanism employed by FHV protein A to initiate RNA synthesis has not been determined. In this study, we characterized the RdRP activity of FHV protein A in detail and revealed that it can initiate RNA synthesis via a de novo (primer-independent) mechanism. Moreover, we found that FHV protein A also possesses a terminal nucleotidyl transferase (TNTase) activity, which was able to restore the nucleotide loss at the 3?-end initiation site of RNA template to rescue RNA synthesis initiation in vitro, and may function as a rescue and protection mechanism to protect the 3? initiation site, and ensure the efficiency and accuracy of viral RNA synthesis. Altogether, our study establishes the de novo initiation mechanism of RdRP and the terminal rescue mechanism of TNTase for FHV protein A, and represents an important advance toward understanding FHV RNA replication. PMID:24466277
Gitlin, Leonid; Hagai, Tzachi; LaBarbera, Anthony; Solovey, Mark; Andino, Raul
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
Parameswaran, V; Ishaq Ahmed, V P; Shukla, Ravi; Bhonde, R R; Sahul Hameed, A S
Two new cell lines, SIMH and SIGE, were derived from the heart of milkfish (Chanos chanos), a euryhaline teleost, and from the eye of grouper (Epinephelus coioides), respectively. These cell lines were maintained in Leibovitz's L-15 supplemented with 20% fetal bovine serum (FBS). The SIMH cell line was subcultured more than 50 times over a period of 210 days and SIGE cell line has been subcultured 100 times over a period of 1 1/2 years. The SIMH cell line consists predominantly of fibroblastic-like cells. The SIGE cell line consists predominantly of epithelial cells. Both the cell lines were able to grow at temperatures between 25 and 32 degrees C with an optimum temperature of 28 degrees C. The growth rate of these cells increased as the proportion of FBS increased from 2% to 20% at 28 degrees C with optimum growth at the concentrations of 15% or 20% FBS. Seven marine fish viruses were tested to determine the susceptibility of these cell lines. The SIGE cell line was found to be susceptible to nodavirus, MABV NC-1 and Y6, and the infection was confirmed by cytopathic effect (CPE) and reverse transcriptase-polymerase chain reaction. When these cells were transfected with pEGFP-N1 vector DNA, significant fluorescent signals were observed, suggesting that these cell lines can be a useful tool for transgenic and genetic manipulation studies. Further, these cell lines are characterized by immunocytochemistry using confocal laser scanning microscopy (CFLSM). PMID:17216384
Onions, D; Côté, C; Love, B; Toms, B; Koduri, S; Armstrong, A; Chang, A; Kolman, J
Massively parallel, deep, sequencing of the transcriptome coupled with algorithmic analysis to identify adventitious agents (MP-Seq™) is an important adjunct in ensuring the safety of cells used in vaccine production. Such cells may harbour novel viruses whose sequences are unknown or latent viruses that are only expressed following stress to the cells. MP-Seq is an unbiased and comprehensive method to identify such viruses and other adventitious agents without prior knowledge of the nature of those agents. Here we demonstrate its utility as part of an integrated approach to identify and characterise potential contaminants within commonly used virus and vaccine production cell lines. Through this analysis, in combination with more traditional approaches, we have excluded the presence of porcine circoviruses in the ATCC Vero cell bank (CCL-81), however, we found that a full length betaretrovirus related to SRV can be expressed in these cells, a factor that may be of importance in the production of certain vaccines. Similarly, insect cells are proving to be valuable for the production of virus like particles and sub-unit vaccines, but they can harbour a range of latent viruses. We show that following MP-Seq of the Trichoplusia ni (High Five cell line) transcriptome we were able to detect a contaminating, latent nodavirus and identify an expressed errantivirus genome. Collectively, these studies have reinforced the role of MP-Seq as an integral tool for the identification of contaminating agents in vaccine cell substrates. PMID:21651935
Luo, Yu-Chun; Wang, Chun-Hsiung; Wu, Yi-Min; Liu, Wangta; Lu, Ming-Wei; Lin, Chan-Shing
Dragon grouper nervous necrosis virus (DGNNV), a member of the genus Betanodavirus, causes high mortality of larvae and juveniles of the grouper fish Epinephelus lanceolatus. Currently, there is no reported crystal structure of a fish nodavirus. The DGNNV virion capsid is derived from a single open reading frame that encodes a 338-amino-acid protein of approximately 37?kDa. The capsid protein of DGNNV was expressed to form virus-like particles (VLPs) in Escherichia coli. The VLP shape is T = 3 quasi-symmetric with a diameter of ?38?nm in cryo-electron microscopy images and is highly similar to the native virion. In this report, crystals of DGNNV VLPs were grown to a size of 0.27?mm within two weeks by the hanging-drop vapour-diffusion method at 283?K and diffracted X-rays to ?7.5?Å resolution. In-house X-ray diffraction data of the DGNNV VLP crystals showed that the crystals belonged to space group R32, with unit-cell parameters a = b = 353.00, c = 800.40?Å, ? = ? = 90, ? = 120°. 23?268 unique reflections were acquired with an overall Rmerge of 18.2% and a completeness of 93.2%. Self-rotation function maps confirmed the fivefold, threefold and twofold symmetries of the icosahedron of DGNNV VLPs. PMID:25084387
Ruelas-Inzunza, J; Green-Ruiz, C; Zavala-Nevárez, M; Soto-Jiménez, M
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 ?g g ?¹ dry weight) and Cr (0.01 ?g g?¹) were detected during the dry season in Gobiesox fluviatilis and Agonostomus monticola, respectively; the highest levels of Hg (0.56 ?g g?¹) were detected during the dry season in Guavina guavina and Mugil curema. In relation to Pb, the highest level (1.65 ?g g?¹) was detected in A. monticola during the dry season. In crustaceans, highest levels of Cd (0.05 ?g g?¹) occurred in Macrobrachium occidentale during both seasons; highest concentration of Cr (0.09 ?g g?¹) was also detected in M. occidentale during the dry season. With respect to Hg, highest level (0.20 ?g g?¹) was detected during the rainy season in Macrobrachium americanum; for Pb, the highest concentration (2.4 ?g 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
Scapigliati, G; Buonocore, F; Randelli, E; Casani, D; Meloni, S; Zarletti, G; Tiberi, M; Pietretti, D; Boschi, I; Manchado, M; Martin-Antonio, B; Jimenez-Cantizano, R; Bovo, G; Borghesan, F; Lorenzen, N; Einer-Jensen, K; Adams, S; Thompson, K; Alonso, C; Bejar, J; Cano, I; Borrego, J J; Alvarez, M C
Naïve sea bass juveniles (38.4 + or - 4.5 g) were intramuscularly infected with a sublethal dose of betanodavirus isolate 378/I03, followed after 43 days by a similar boosting. This infection resulted in an overall mortality of 7.6%. At various intervals, sampling of fish tissues was performed to investigate: i) B and T lymphocyte content in organs and tissues; ii), proliferation of leucocytes re-stimulated in vitro with inactivated virus; iii) presence of serum antibody specific for betanodavirus; iv) expression of genes coding for the following immunoregulatory molecules involved in innate and acquired responses: type I IFN, Mx, IL-1, Cox-2; IL-10, TGF-beta, TCRbeta, CD4, CD8alpha, IgM, by using a quantitative PCR array system developed for sea bass. The obtained results showed a detectable increase of T cells and B cells in PBL during betanodavirus infection. Furthermore, leucocytes obtained from blood, head kidney, and gills showed a detectable "in vitro" increase in viability upon addition of inactivated viral particles, as determined by measuring intracellular ATP concentration. ELISA analysis of sera showed that exposure to nodavirus induced a low, but specific antibody titer measured 43 days after infection, despite the presence of measurable levels of natural antibody. Finally, a strong upregulation of genes coding for type I IFN, Mx, and IgM was identified after both infection and boosting. Interestingly, an upregulation of Cox-2 until boosting, and of TGF-beta and IL-10 after boosting was also observed, while the other tested genes did not show any significant variations with respect to mock-treated fish. Overall, our work represents a first comprehensive analysis of cellular and molecular immune parameters in a fish species exposed to a pathogenic virus. PMID:19925869
Buonocore, Francesco; Randelli, Elisa; Tranfa, Paola; Scapigliati, Giuseppe
The CD83 cell surface marker is an important and intriguing component of immune system. It is considered the best marker for mature human dendritic cells, but it is also important for thymic development of T cells, and it also plays a role as a regulator of peripheral B-cell function and homeostasis. A CD83-like molecule was identified in sea bass (Dicentrarchus labrax) by EST sequencing of a thymus cDNA library; the CD83 cDNA is composed of 816 bp and the mature CD83 peptide consists of 195 amino acids, with a putative signal peptide of 18 amino acids and two possible N-glycosylation sites. The comparison of sea bass CD83 sequence with its homologues in other fish species and mammals shows some differences, with two cysteine residues conserved from fish to mammals and a high variability both in the total number of cysteines and in mature CD83 sequence polypeptide length. Basal transcripts levels of CD83 mRNA are highest in liver, followed by thymus. The in vitro treatment of head kidney leukocytes with LPS resulted in a down-regulation on CD83 mRNA leves both after 4 and 24 h, whereas with poly I:C an up-regulation after 4h followed by a down-regulation at 24 h was observed. An in vivo infection of sea bass juveniles with nodavirus induced an increase of CD83 expression on head kidney leukocytes both after 6 and 24 h and a decrease after 72 h. On the other hand, an in vivo infection with Photobacterium damselae bacteria induced a decrease of CD83 transcript levels after 6 and 24 h and an increase after 72 h. These findings suggest in sea bass CD83 expression could be modulated by viral and bacterial immune response. PMID:22554578
Venter, P Arno; Marshall, Dawn; Schneemann, Anette
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
Background Nodaviridae is a family of non-enveloped isometric viruses with bipartite positive-sense RNA genomes. The Nodaviridae family consists of two genera: alpha- and beta-nodavirus. Alphanodaviruses usually infect insect cells. Some commercially available insect cell lines have been latently infected by Alphanodaviruses. Results A non-enveloped small virus of approximately 30 nm in diameter was discovered co-existing with a recombinant Helicoverpa armigera single nucleopolyhedrovirus (HearNPV) in Hz-AM1 cells. Genome sequencing and phylogenetic assays indicate that this novel virus belongs to the genus of alphanodavirus in the family Nodaviridae and was designated HzNV. HzNV possesses a RNA genome that contains two segments. RNA1 is 3038 nt long and encodes a 110 kDa viral protein termed protein A. The 1404 nt long RNA2 encodes a 44 kDa protein, which exhibits a high homology with coat protein precursors of other alphanodaviruses. HzNV virions were located in the cytoplasm, in association with cytoplasmic membrane structures. The host susceptibility test demonstrated that HzNV was able to infect various cell lines ranging from insect cells to mammalian cells. However, only Hz-AM1 appeared to be fully permissive for HzNV, as the mature viral coat protein essential for HzNV particle formation was limited to Hz-AM1 cells. Conclusion A novel alphanodavirus, which is 30 nm in diameter and with a limited host range, was discovered in Hz-AM1 cells. PMID:21682922
Huang, Xiaohong; Huang, Youhua; Cai, Jia; Wei, Shina; Ouyang, Zhengliang; Qin, Qiwei
Interferon-stimulated gene 15 (ISG15) is an ubiquitin homolog that is significantly induced by type I interferons or viral infections. Groupers, Epinephelus spp. being maricultured in China and Southeast Asian countries, always suffer from virus infection, including iridovirus and nodavirus. To date, the roles of grouper genes, especially interferon related genes in virus infection remained largely unknown. Here, the ISG15 homolog (EcISG15) was cloned from grouper Epinephelus coioides and its immune response to Singapore grouper iridovirus (SGIV) and grouper nervous necrosis virus (GNNV) was investigated. The full-length EcISG15 cDNA was composed of 948 bp and encoded a polypeptide of 155 amino acids with 37-68% identity with the known ISG15 homologs from other fish species. Amino acid alignment analysis indicated that EcISG15 contained two ubiquitin-like (UBL) domains and an Ub-conjugation domain (LRGG). Expressional analysis showed that EcISG15 was dramatically induced by GNNV infection, poly I:C or poly dA-dT treatment, but no obvious changes were observed during SGIV infection. Immunofluorescence assay showed that EcISG15 localized mainly in the cytoplasm of grouper cells in response to poly I:C stimulation or GNNV infection, but not in mock or SGIV infected cells. Western blot analysis indicated that the ISGylation was absent in SGIV-infected cells, but significantly enhanced in GNNV-infected or poly I:C transfected cells, suggesting that EcISG15 might play different roles in SGIV and GNNV infection. Furthermore, overexpression of EcISG15 in vitro inhibited the transcription of GNNV genes significantly. Taken together, the results indicated that fish ISG15 might exert important roles against RNA virus infection. PMID:23403156
Maier, Helena J.; Hawes, Philippa C.; Cottam, Eleanor M.; Mantell, Judith; Verkade, Paul; Monaghan, Paul; Wileman, Tom; Britton, Paul
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
Johnson, Karyn N.; Zeddam, Jean-Louis; Ball, L. Andrew
Pariacoto virus (PaV) was recently isolated in Peru from the Southern armyworm (Spodoptera eridania). PaV particles are isometric, nonenveloped, and about 30 nm in diameter. The virus has a bipartite RNA genome and a single major capsid protein with a molecular mass of 39.0 kDa, features that support its classification as a Nodavirus. As such, PaV is the first Alphanodavirus to have been isolated from outside Australasia. Here we report that PaV replicates in wax moth larvae and that PaV genomic RNAs replicate when transfected into cultured baby hamster kidney cells. The complete nucleotide sequences of both segments of the bipartite RNA genome were determined. The larger genome segment, RNA1, is 3,011 nucleotides long and contains a 973-amino-acid open reading frame (ORF) encoding protein A, the viral contribution to the RNA replicase. During replication, a 414-nucleotide long subgenomic RNA (RNA3) is synthesized which is coterminal with the 3? end of RNA1. RNA3 contains a small ORF which could encode a protein of 90 amino acids similar to the B2 protein of other alphanodaviruses. RNA2 contains 1,311 nucleotides and encodes the 401 amino acids of the capsid protein precursor ?. The amino acid sequences of the PaV capsid protein and the replicase subunit share 41 and 26% identity with homologous proteins of Flock house virus, the best characterized of the alphanodaviruses. These and other sequence comparisons indicate that PaV is evolutionarily the most distant of the alphanodaviruses described to date, consistent with its novel geographic origin. Although the PaV capsid precursor is cleaved into the two mature capsid proteins ? and ?, the amino acid sequence at the cleavage site, which is Asn/Ala in all other alphanodaviruses, is Asn/Ser in PaV. To facilitate the investigation of PaV replication in cultured cells, we constructed plasmids that transcribed full-length PaV RNAs with authentic 5? and 3? termini. Transcription of these plasmids in cells recreated the replication of PaV RNA1 and RNA2, synthesis of subgenomic RNA3, and translation of viral proteins A and ?. PMID:10799587
Wong, M. H.; Hung, K. M.; Chiu, S. T.
This project investigated the feasibility of using sewage sludge to culture microalgae ( Chlorella-HKBU) and their subsequent usage as feeds for rearing different organisms. Part II of the project evaluated the results of applying the sludge-grown algae to feed Oreochromis mossambicus (fish), Macrobrachium hainenese (shrimp), and Moina macrocopa (cladocera). In general, the yields of the cultivated organisms were unsatisfactory when they were fed the sludge-grown algae directly. The body weights of O. mossambicus and M. macrocopa dropped 21% and 37%, respectively, although there was a slight increase (4.4%) in M. hainenese. However, when feeding the algal-fed cladocerans to fish and shrimp, the body weights of the fish and shrimp were increased 7% and 11% accordingly. Protein contents of the cultivated organisms were comparable to the control diet, although they contained a rather high amount of heavy metals. When comparing absolute heavy metal contents in the cultivated organisms, the following order was observed: alga > cladocera > shrimp, fish > sludge extracts. Bioelimination of heavy metals may account for the decreasing heavy metal concentrations in higher trophic organisms.
Wong; Hung; Chiu
This project investigated the feasibility of using sewage sludge to culture microalgae (Chlorella-HKBU) and their subsequent usage as feeds for rearing different organisms. Part II of the project evaluated the results of applying the sludge-grown algae to feed Oreochromis mossambicus (fish), Macrobrachium hainenese (shrimp), and Moina macrocopa (cladocera). In general, the yields of the cultivated organisms were unsatisfactory when they were fed the sludge-grown algae directly. The body weights of O. mossambicus and M. macrocopa dropped 21% and 37%, respectively, although there was a slight increase (4.4%) in M. hainenese. However, when feeding the algal-fed cladocerans to fish and shrimp, the body weights of the fish and shrimp were increased 7% and 11% accordingly. Protein contents of the cultivated organisms were comparable to the control diet, although they contained a rather high amount of heavy metals. When comparing absolute heavy metal contents in the cultivated organisms, the following order was observed: alga > cladocera > shrimp, fish > sludge extracts. Bioelimination of heavy metals may account for the decreasing heavy metal concentrations in higher trophic organisms. PMID:8661608
Shuhaimi-Othman, M; Nadzifah, Y; Nur-Amalina, R; Umirah, N S
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 LC(50) 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 ?g 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 ?g?L(-1) and 9.0, 2.0, 19.9, and 36.7 ?g?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 LC(50) 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
Gutierrez, María Florencia; Negro, Carlos Leandro
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
Deep, Kamal; Poddar, Abhijit; Das, Subrata K
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
Lucena, Teresa; Ruvira, María A; Pascual, Javier; Garay, Esperanza; Macián, M Carmen; Arahal, David R; Pujalte, María J
A facultatively anaerobic marine gammaproteobacterium, designated strain M46(T), was isolated from Mediterranean seawater at Malvarrosa beach, Valencia, Spain. The strain was characterized by using a polyphasic approach and was found to be situated within the genus Photobacterium in the family Vibrionaceae. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain M46(T) was closely related to P. rosenbergii CECT 7644(T), P. halotolerans CECT 5860(T) and P. ganghwense CECT 7641(T), showing sequence similarities of 96.8, 96.4 and 96.2?%, respectively. According to the results of phylogenetic analyses based on recA and gyrB gene sequences, the most closely related taxon was P. ganghwense CECT 7641(T) with 87.4 and 85.0?% sequence similarity, respectively. Regardless of the gene used in phylogenetic analysis, strain M46(T) always formed a separate and stable clade containing these three species of the genus Photobacterium. Strain M46(T) was not luminescent and produced a diffusible brown pigment. It required NaCl to grow, reduced nitrate to nitrite and oxidized a small number of substrates in Biolog GN plates. Strain M46(T) was positive for arginine dihydrolase (ADH), ?-galactosidase, aesculin hydrolysis and DNase activity. In API ZYM tests, the novel strain was positive for alkaline phosphatase, leucine arylamidase and acidic phosphatase activities. The major cellular fatty acids were unsaturated C(18) and C(16), as in other members of the genus Photobacterium, but their relative amounts and the presence or absence of other fatty acids differentiated strain M46(T) from its closest relatives. Based on the results of this polyphasic taxonomic study, strain M46(T) represents a novel species of the genus Photobacterium, for which the name Photobacterium aphoticum is proposed. The type strain is M46(T) (?=?CECT 7614(T) ?=?KCTC 23057(T)). PMID:20675441
MacKenzie, R. A.; Cormier, N.
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.
Santos, L C F; Belli, N M; Augusto, A; Masui, D C; Leone, F A; McNamara, J C; Furriel, R P M
To better comprehend physiological adaptation to dilute media and the molecular mechanisms underlying ammonia excretion in palaemonid shrimps, we characterized the (Na+,K+)-ATPase from Macrobrachium amazonicum gills, disclosing high- (K(0.5) = 4.2+/-0.2 micromol L(-1); V = 33.9+/-1.9 U mg(-1)) and low-affinity (K(0.5) = 0.144+/-0.010 mmol L(-1); V = 232.9+/-15.3 U mg(-1)) ATP hydrolyzing sites. Stimulation by Na+ (K(0.5) = 5.5+/-0.3 mmol L(-1); V = 275.1+/-15.1 U mg(-1)), Mg2+ (K(0.5) = 0.79+/-0.06 mmol L(-1); V = 261.9+/-18.3 U mg(-1)), K+ (K(M) = 0.88+/-0.04 mmol L(-1); V = 271.8+/-10.9 U mg(-1)) and NH4(+) (K(M) = 5.0+/-0.2 mmol L(-1); V = 385.9+/-15.8 U mg(-1)) obeys single saturation curves, activity being stimulated synergistically by NH4(+) and K+. There is a single K+ binding site, NH4(+) binding to a second, exclusive site, stimulating activity by 33%, modulating K+ affinity. (Na+,K+)-ATPase activity constitutes approximately 80% of total ATPase activity (K(Iouabain) = 147.5+/-8.9 micromol L(-1)); Na+-, K+-, Ca2+-, V- and F(o)F(1)-ATPases are also present. M. amazonicum microsomal fractions possess approximately 2-fold less (Na+,K+)-ATPase alpha-subunit than M. olfersi, consistent with a 2.6-fold lower specific activity. These differences in (Na+, K+)-ATPase stimulation by ATP and ions, and specific activities of other ATPases, suggest the presence of distinct biochemical adaptations to life in fresh water in these related species. PMID:17521934
Marbaniang, Deswyn G; Poddar, Raj K; Nongkynrih, Phlis; Khathing, Darlando T
The study was performed using a silicon surface barrier alpha spectrometer at Bhabha Atomic Research Centre, Mumbai, India. Through the study, the observed (210)Po activity in water sample from different locations in the Domiasiat area ranges from 0.04 to 0.69 Bq/l. The daily and annual intake of (210)Po through water was also estimated and the mean value of 0.72 and 263.61 Bq, respectively, were observed. It is observed that the effective doses through water were higher than the World Health Organization recommended dose of 0.05 mSv/year. The total annual effective doses through terrestrial ingestion for all the locations was studied and the mean annual effective dose was observed to be 0.315 mSv, which, when compared to the worldwide and the Indian values, was observed to be slightly higher. The mean activity in soil is found to be 124.8 +/-5.7 Bq/kg and in meat the activity is 0.43 +/-0.05 Bq/kg. In fishes, an activity of 0.48 +/-0.07 Bq/kg in Garra lamta, 0.29 +/-0.02 Bq/kg in Neolissocheilus hexaganolepis, and 3.3 +/-0.1 Bq/kg in Macrobrachium sp. is observed. Activity concentration in plant samples was analyzed and the activity ranges from 0.020 +/-0.002 to 9.69 +/-0.35 Bq/kg. Committed effective dose by the adult population of the Domiasiat area through intake of (210)Po through these food items was also determined and compared with the Indian average value and the worldwide average value. PMID:19242810
Li, Fajun; Bai, Hongkun; Zhang, Wenyi; Fu, Hongtuo; Jiang, Fengwei; Liang, Guoxia; Jin, Shubo; Sun, Shengming; Qiao, Hui
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
Song, Lirong; Chen, Wei; Peng, Liang; Wan, Neng; Gan, Nanqin; Zhang, Xiaoming
For the purpose of understanding the environmental fate of microcystins (MCs) and the potential health risks caused by toxic cyanobacterial blooms in Lake Taihu, a systematic investigation was carried out from February 2005 to January 2006. The distribution of MCs in the water column, and toxin bioaccumulations in aquatic organisms were surveyed. The results suggested that Lake Taihu is heavily polluted during summer months by toxic cyanobacterial blooms (with a maximum biovolume of 6.7 x 10(8)cells/L) and MCs. The maximum concentration of cell-bound toxins was 1.81 mg/g (DW) and the dissolved MCs reached a maximum level of 6.69 microg/L. Dissolved MCs were always found in the entire water column at all sampling sites throughout the year. Our results emphasized the need for tracking MCs not only in the entire water column but also at the interface between water and sediment. Seasonal changes of MC concentrations in four species of hydrophytes (Eichhornic crassipes, Potamogeton maackianus, Alternanthera philoxeroides and Myriophyllum spicatum) ranged from 129 to 1317, 147 to 1534, 169 to 3945 and 124 to 956 ng/g (DW), respectively. Toxin accumulations in four aquatic species (Carassius auratus auratu, Macrobrachium nipponensis, Bellamya aeruginosa and Cristaria plicata) were also analyzed. Maximum toxin concentrations in the edible organs and non-edible visceral organs ranged from 378 to 730 and 754 to 3629 ng/g (DW), respectively. Based on field studies in Lake Taihu, risk assessments were carried out, taking into account the WHO guidelines and the tolerable daily intake (TDI) for MCs. Our findings suggest that the third largest lake in China poses serious health threats when serving as a source of drinking water and for recreational use. In addition, it is likely to be unsafe to consume aquatic species harvested in Lake Taihu due to the high-concentrations of accumulated MCs. PMID:17537477
Watanabe, Karen H; Desimone, Frank W; Thiyagarajah, Arunthavarani; Hartley, William R; Hindrichs, Albert E
Between 1990 and 1994, samples of three shellfish species (i.e. blue crab, Callinectes sapidus;crayfish, Procambarus acutis; and river shrimp, Macrobrachium ohionii) and 16 fish species and were collected at six sites along the lower Mississippi River by the Louisiana Department of Environmental Quality, Office of Water Resources in coordination with the US Environmental Protection Agency. The fish species included: bigmouth buffalo (Ictiobus cyanellus); blue catfish (Ictalurus furcatus); carp (Cyprinus carpio); channel catfish (Ictalurus punctatus); cobia (Rachycentron canadum); flathead catfish (Pylodictis olivaris); freshwater drum (Aplodinotus grunniens); largemouth bass (Micropterus salmoides); long nose gar (Lepisosteus osseus); red drum (Sciaenops ocellatus); red snapper (Lutjanus campechanus); smallmouth buffalo (Ictiobus bubalus); spotted gar (Lepisosteus oculatus); striped bass (Morone saxatilis); white bass (Morone chrysops); and white crappie (Pomoxis annularis). Organic compound and heavy metal concentrations were measured in 161 composite fish tissue samples where each composite included three to 10 individual fish. Nineteen chemicals, found at measurable levels in sample tissues, were used in calculations of lifetime excess cancer and non-cancer risks due to fish consumption. We calculated: 574 chemical-specific cancer risks; 41 total cancer risks; and 697 margins of exposure based on a consumption rate of one 8-ounce meal per week (0.032 kg/day), a body weight of 70 kg and reported cancer potency factors and reference doses. We identified nine species of concern (blue catfish, carp, channel catfish, cobia, crayfish, flathead catfish, red drum, spotted gar and striped bass) based on total cancer risk greater than 10(-4) or margin of exposure greater than 1, and whether or not samples collected in subsequent years resulted in lower risks. The compounds primarily responsible for the elevated risks were aldrin, dieldrin, alpha-benzene hexachloride, gamma-benzene hexachloride, heptachlor epoxide, arsenic and mercury. PMID:12526903
Adeogun, Aina O; Ibor, Oju R; Omogbemi, Emmanuel D; Chukwuka, Azubuike V; Adegbola, Rachel A; Adewuyi, Gregory A; Arukwe, Augustine
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
Zhang, Guizhai; Pan, Zhaoke; Wang, Xiaoming; Mo, Xiaojie; Li, Xiaoming
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