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At least 847 records · Page 47Linked to original sources
PubMed2002

A new look at an old visual system: structure and development of the compound eyes and optic ganglia of the brine shrimp Artemia salina Linnaeus, 1758 (Branchiopoda, anostraca).

Compared to research carried out on decapod crustaceans, the development of the visual system in representatives of the entomostracan crustaceans is poorly understood. However, the structural evolution of the arthropod visual system is an important topic in the new debate on arthropod relationships, and entomostracan crustaceans play a key role in this discussion. Hence, data on structure and ontogeny of the entomostracan visual system are likely to contribute new aspects to our understanding of arthropod phylogeny. Therefore, we explored the proliferation of neuronal stem cells (in vivo incorporation of bromodeoxyuridine) and the developmental expression of synaptic proteins (immunohistochemistry against synapsins) in the developing optic neuropils of the brine shrimp Artemia salina Linnaeus, 1758 (Crustacea, Entomostraca, Branchiopoda, Anostraca) from hatching to adulthood. The morphology of the adult visual system was examined in serial sections of plastic embedded specimens. Our results indicate that the cellular material that gives rise to the visual system (compound eyes and two optic ganglia) is contributed by the mitotic activity of neuronal stem cells that are arranged in three band-shaped proliferation zones. Synapsin-like immunoreactivity in the lamina ganglionaris and the medulla externa initiated only after the anlagen of the compound eyes had already formed, suggesting that the emergence of the two optic neuropils lags behind the proliferative action of these stem cells. Neurogenesis in A. salina is compared to similar processes in malacostracan crustaceans and possible phylogenetic implications are discussed.

Age Factors↗
PubMed1982

A comparative study by serial electron microscopy of neuromuscular junctions in the dimorphic claws of the snapping shrimp, Alpheus heterochelis.

We compared the neuromuscular junctions on the main closer muscle in the first pair of chelipeds in the snapping shrimp Alpheus heterochelis by serial section electron microscopy. We sought an ultrastructural basis for the different behavioral and physiological functions of these dimorphic claws and for the role of the nervous system in claw transformation. We were unable to detect any statistically significant morphological differences between the junctions. Further, we found the muscle fiber populations and filament arrangements, as well as the electrical properties of the fibers, to be more homogeneous and similar to each other in A. heterochelis than those reported for another species, A. armillatus. We consider our results in light of recent data on the anatomy and electrical properties of the motor neurons within the CNS and conclude that the neural trigger for claw transformation involves factors not revealed by conventional electron microscopy.

Animals↗
PubMed1988

Disruption of muscle reorganization by lesions of the peripheral nerve in transforming claws of snapping shrimps.

We have performed surgical transections on nerves in the transforming claws of snapping shrimps. In normal transformation muscle restructuring occurs, involving degeneration of some fibers and biochemical changes in others. Surgical section of the entire second limb nerve root or of its distal, dorsal branch--both of which contain the motor axons to the closer muscle--prevents muscle restructuring, even though transformation of external claw morphology proceeds. Furthermore, nerve lesions must be performed within a specific time period after transformation has been triggered in order for the effects to be observed. We suggest that transformation involves an early sensitization of the targeted muscle and that this process depends upon an intact nervous pathway within the second nerve root.

Action Potentials↗
PubMed2004

Identification of structural proteins from shrimp white spot syndrome virus (WSSV) by 2DE-MS.

White spot syndrome virus (WSSV) is a major shrimp pathogen that also infects many other species of crustaceans. Its 305-kb double-stranded DNA genome has the capacity to encode 181 presumptive proteins. In an attempt to identify the viral proteins from the 181 theoretical proteins, proteins of the purified WSSV were separated by two-dimensional electrophoresis (2-DE). More than 60 protein spots were revealed, as detected by silver staining, from which 12 viral proteins were identified by mass spectrometry. In total, 25 WSSV proteins, including those reported in one of our earlier studies (Huang et al., Mol Cell Proteomics 2002;1:223-231), were revealed by this proteomic approach, and their corresponding genes were further confirmed by reverse transcription-polymerase chain reaction (RT-PCR). Two of them were characterized to be WSSV envelope proteins using immuno-electron microscopy. Our study showed that the proteomic approach is a powerful method for discovering the viral structural proteins and their corresponding genes.

Animals↗
PubMed2002

Brine shrimp lethality assay of Bacopa monnieri.

Successive petroleum ether, chloroform, ethanol and water extracts, a saponin rich fraction (SRF) and bacoside A isolated from Bacopa monnieri were tested for brine shrimp lethality. Successive ethanol extracts and SRF showed potent activity. Bacoside A showed the maximum activity with a LC(50) of 38.3 microg/mL. The results confirmed the previous reports of an anticancer effect of Bacopa monnieri and suggest bacoside A as the active constituent.

Animals↗
PubMed2005

Trophic transfer of cyanobacterial toxins from zooplankton to planktivores: consequences for pike larvae and mysid shrimps.

The aim of this study was to evaluate the potentially harmful effects of zooplankton preexposed to cyanobacteria on two planktivorous animals: a fish larva (pike, Esox lucius) and a mysid shrimp (Neomysis integer). The planktivores were fed zooplankton from a natural community that had been preexposed to cell-free extract or to purified toxin (nodularin) of the cyanobacterium Nodularia spumigena, and the growth, feeding, and pellet production of the planktivores, as well as the toxin content of the pellets, were measured. In addition, radiolabeled nodularin ((3)H-dihydronodularin) was used in separate experiments to measure the vector transfer of nodularin from zooplankton to their predators. During 11-day exposures, dissolved nodularin was transferred to pike larvae and N. integer via zooplankton at very low rates of accumulation. Treatment with N. spumigena extract decreased the ingestion and feces production rates of pike larvae. With purified nodularin alone, no such effect could be observed. No effect on molting cycle length, fecal pellet production, C:N ratio, or growth of N. integer was detected. The results suggest that dissolved cyanobacterial toxins released during bloom decay can have a negative impact on feeding and, hence, on the growth of fish larvae via zooplankton, even without direct contact between cyanobacteria and the fish.

Animals↗
PubMed2001

Proteinase activity in the white shrimp (Penaeus vannamei) clotting protein.

The clottable protein (CP) was isolated from white shrimp, Penaeus vannamei plasma as a 400-kDa protein that splits to two identical 200-kDa subunits when it is reduced with 2-ME. However, using DTT as reducing agent, four main bands were observed; two of them (179 and 125 kDa) had the same N-terminus sequence of the intact CP, indicating that most fragmentation occurs in the carboxy-terminus. The proteinase activity of reduced CP was detected using azoalbumin as substrate. Proteinase activity was only detected in the reduced, but not alkylated protein. Trypsin and papain, as well as soybean trypsin inhibitor and E64, were included for comparison. Proteolytic activity of reduced CP was inhibited by E64, but not by STI, indicating that such activity corresponds to a cysteine type proteinase.

Animals↗
PubMed1996

Effect of ammonia on survival and adenylate energy charge in the shrimp Palaemonetes varians.

Shrimps Palaemonetes varians were submitted to lethal and sublethal concentrations of ammonia in order to measure lethality, ATP and adenylate nucleotide levels, and adenylate energy charge (AEC) index. LC50 was of about 3 mg/liter of ammonia. Adenylate measurements were performed over a period of 14 days and for two different concentrations. A population submitted to 0.5 mg/liter of ammonia exhibited high survival and a marked consumption of ATP, whereas high mortality and disordered ATP metabolism are the characteristics of the population submitted to 3 mg/liter of ammonia. An homeostatic model was applied in order to explain the significance of the AEC signal. Here, AEC is proposed as a measure of the limits of active response of an organism to environmental stress.

Adenosine Diphosphate↗
PubMed2000

Characterisation of different morphological features of black tiger shrimp (Penaeus monodon) haemocytes using monoclonal antibodies.

Monoclonal antibodies (mabs) specific for Penaeus monodon haemocytes were produced by immunising mice with membrane lysates of shrimp haemocytes. Four mabs (WSH 6, WSH 7, WSH 8 and WSH 16) were characterised using flow cytometry, light microscopy, laser scanning microscopy, electron microscopy and immunoprecipitation. WSH 6 recognised a carbohydrate determinant on an 85 kDa molecule. WSH 7, WSH 8 and WSH 16 recognised 50, 35 and 115 kDa molecules, respectively. For all mabs, differences in amount and intensity of the labelling were found when haemocytes were fixed immediately in 2% formaldehyde in Alsever's Solution (AS), compared with non-fixed haemocytes that were kept in AS (which reduced activation of the haemocytes) or in L15 cell culture medium. WSH 6 reacted with the cell membranes of all fixed haemocytes, while WSH 7 and WSH 16 reacted with the cell membranes of >80% of fixed haemocytes. The membrane labelling appeared to decrease when cells were kept in L15 medium. WSH 8 did not react with the haemocyte membranes. All mabs reacted with some granules, mainly present in the hyaline cells, when the haemocytes were immediately fixed. When non-fixed cells were kept in AS and in L15 medium, positive granules were also observed in semigranular and granular haemocytes as well as in the largest granules of a fourth cell type, that contains many granules of different size and electron density. Immunoreactive extracellular thread-like material could be observed in cells in L15 medium. The change in staining pattern was extreme for WSH 8, somewhat less for WSH 6 and WSH 7 and the lowest for WSH 16. Double labelling revealed that all mabs showed a different staining pattern on membranes as well as on granules. WSH 16 also showed labelling in cytoplasmic vesicles, as well as in haemolymph plasma on histological sections. The hypothesis is put forward that immunoreactive molecules recognised by these mabs, are related to haemocyte activation factors.

Animals↗
PubMed2002

The role of the haematopoietic tissue in haemocyte production and maturation in the black tiger shrimp (Penaeus monodon).

The haematopoietic tissue (HPT) of the black tiger shrimp (Penaeus monodon) is located in different areas in the cephalothorax, mainly at the dorsal side of the stomach and in the onset of the maxillipeds and, to a lesser extent, towards the antennal gland. In young and in experimentally stimulated animals, the HPT is expanded in relatively larger and more numerous lobules throughout the cephalothorax. Four cell types could be identified in the HPT by electron microscopy. The type 1 cells are the presumed precursor cells that give rise to a large- and a small-granular young haemocyte, denominated as the type 2 and type 3 cells, respectively. A gradient of maturation from the type 1 towards the type 2 or 3 cells could frequently be observed. The presumed precursor cells are located towards the exterior of the lobules and maturing young haemocytes towards the inner part, where they can be released into the haemal lacunae. The type 4 cells show typical features of interstitial cells. Different stimulation experiments were carried out and various techniques were used to study the HPT in relation to the (circulating) haemocytes. The majority of the cells in the HPT are able to proliferate and proliferation can be increased significantly after the injection of saline and, to a much higher extent, after LPS injection. The circulating haemocytes of crustaceans are generally divided into hyaline (H), semigranular (SG) or granular (G) cells, of which large- and small-granular variants of each of these were suggested in the present study. Even after stimulation in this study, the circulating haemocytes scarcely divide. The high variations that were found in the total haemocyte count in the stimulation experiments were not accompanied by significant differences in differential haemocyte count and, therefore, appeared to be a less useful indicator of stress or health in P. monodon. Light and electron microscopical observations support the regulation of the populations of the different haemocyte types in the circulation by (stored) haemocytes from the connective tissue. In conclusion, according to morphological and immuno-chemical criteria, it is proposed in the present study to divide the haemocytes into a large-and a small-granular developmental series. After extensive morphological observations, it is suggested that the hyaline cells are the young and immature haemocytes of both the large- and small-granular cell line that are produced in the HPT, and can be released into the haemolymph. Indications were found that the granular cells, of at least the large-granular cell line, mature and accumulate in the connective tissue and are easily released into the haemolymph. Combining the results of the present study with literature, this proposed model for haemocyte proliferation, maturation and reaction will be discussed.

Animals↗
PubMed2002

Different bacterial lipopolysaccharides as toxicants and stressors in the shrimp Palaemon elegans.

This study compares the in vivo haemocytic response of shrimp, Palaemon elegans (Rathke) to different types of LPS injection. In particular it investigates to what degree and speed the haemocytopenia varies between LPSs from different sources. It further compares the tolerated doses of different LPSs in these animals and finds substantial differences in the various toxicity types. The work then relates this to blood glucose levels and stress-linked variations in glycaemic status. The order of LPS decreasing toxicity determined by LD50 at 96 h was: Salmonella enteritidis, Serratia marcescens, Pseudomonas aeruginosa 10, Escherichia coli K-235 and E. coli 0111:B4. Eyestalkless animals were more sensitive to LPS. The effects of injected LPS on circulating total blood cell count (THC) was tested. The results show that LPS caused a decrease in THC 8 h after injection and then the THC returned to the initial level and this effect depended on the LPS tested. E. coli K-235 was the most effective in causing haemocytopenia followed by E. coli 0111:B4, S. enteritidis, S. marcescens, and P. aeruginosa 10. Moreover, LPS-induced increases in the blood glucose level and the time and dose related curves of response obtained depended on the type of LPS tested. E. coli K-235 LPS was again the most effective in elevating blood glucose followed by E. coli 0111:B4, S. marcescens, S. enteritidis and then P. aeruginosa 10. No significant hyperglycaemia was induced in eyestalkless animals. An inverse order relationship between toxicity (LD50) and stress responses (hyperglycaemia and THC decrease) may suggest a defensive and adaptive role of the latter in occasional septicaemia.

Adaptation, Physiological↗
PubMed2002

1,3-beta-D glucan binding protein (BGBP) from the white shrimp, Penaeus vannamei, is also a heparin binding protein.

Shrimp BGBP was purified as a 100 kDa glycoprotein by affinity chromatography using immobilised heparin. BGBP bound simple carbohydrates, glycosaminoglycans like heparin sulphate and glycoproteins, but it was unable to agglutinate erythrocytes. Using an ELISA-based microplate assay, it was shown that simple carbohydrates such as n-glucose and D-mannose are competitive inhibitors of heparin sulphate binding to BGBP. Based on these properties BGBP is considered as a new type of heparin binding protein.

Animals↗
PubMed2001

Ultrastructural aspects of a new species, Vavraia mediterranica (Microsporidia, Pleistophoridae), parasite of the French Mediterranean shrimp, Crangon crangon (Crustacea, Decapoda).

The life cycle stages of a new species of the genus Vavraia (Microsporidia, Pleistophoridae), which parasitizes the shrimp Crangon crangon (Crustacea, Decapoda), were examined by light and electron microscopy. This parasite was monomorphic with polysporous sporogony and developed in the skeletal muscle of the host. The multinucleate sporogonial plasmodium divided by plasmotomy and multiple division into uninucleate sporoblasts. All stages were surrounded by a thick and amorphous dense coat external to the plasmalemma. This structure gradually became a merontogenetic sporophorous vacuole (MSV) where the sporonts developed into sporoblasts. The MSV was filled with episporontal granular secretory products and eventually contained up to 50 uninucleate spores. During spore morphogenesis, these episporontal granular products within the MSV became organized as episporontal tubular-like structures. In transverse sections, these structures showed a mean diameter of 1.0 microm, but disappeared during the final phase of the spore maturation. Mature spores were ellipsoidal to slightly pyriform and measured 2.30 x 1.41 microm. The polar filament was anisofilar and consisted of a single coil with six to seven turns (rarely five). This new species is named Vavraia mediterranica n. sp.

Animals↗
PubMed1998

Molecular phylogeny and biogeography of the marine shrimp Penaeus.

The evolutionary relationships among 13 species representing all six subgenera of the shrimp genus Penaeus were examined using 558 bp of mitochondrial (mt) DNA from the cytochrome oxidase subunit I gene. Analyses of this sequence revealed high genetic divergence between species (d = 8-24%), a finding which contrasts with previous work, which indicated that genetic diversity, based on electrophoretic analysis of allozymes, was extremely low in Penaeus. Three tree-building methods (maximum parsimony, neighbor joining, and maximum likelihood) were concordant in indicating that current subgenera assignments do not reflect evolutionary partitions within the genus Penaeus. While the molecular phylogenies cast doubt on the validity of subgenera, the observed relationships are concordant with biogeographic boundaries across the tropical range of Penaeus. Both the western Atlantic and eastern Pacific contain monophyletic species pairs which cluster together in all analyses. The Indo-Pacific contains a putative basal taxa (P. indicus), the deepest mtDNA lineages, and the highest diversity, including representatives of all three primary lineages observed in Penaeus. These data are consistent with the suggestion by Dall et al. (1990) that Penaeus arose in the Indo-Pacific and radiated eastward and westward to account for the current circumtropical distribution of the genus. This phylogenetic framework for Penaeus will enhance the scientific foundations for wildlife resource management and breeding experiments (hybridization and related manipulations) designed to improve the commercial value of captive strains.

Animals↗
PubMed2000

Infectious hypodermal and hematopoietic necrosis virus of shrimp is related to mosquito brevidensoviruses.

We purified and sequenced infectious hypodermal and hematopoietic necrosis virus (IHHNV), a small DNA virus of shrimp, from wild Penaeus stylirostris. The virion has a buoyant density of 1.45 as determined by cesium chloride gradient. Analysis of 3873 nucleotides of the viral genome revealed three large open reading frames (ORFs) and parts of the noncoding termini of the viral genome. The left, mid, and right ORFs on the complementary (plus) strand have potential coding capacities of 666 amino acids (aa) (75.77 kDa), 363 aa (42.11 kDa), and 329 aa (37.48 kDa), respectively. The overall genomic organization is similar to that of the mosquito brevidensoviruses. The left ORF most likely encodes the major nonstructural (NS) protein (NS-1) since it contains conserved replication initiator motifs and NTP-binding and helicase domains similar to those in NS-1 from all other parvoviruses. The IHHNV putative NS-1 shares the highest aa sequence homology with the NS-1 of mosquito brevidensoviruses, Aedes densovirus and Aedes albopictus parvovirus. A search for putative splicing sites revealed that the N-terminal region of NS-1 is very likely located in a small ORF upstream of the left ORF. The right ORF is presumed to encode structural polypeptides (VPs), as in other parvoviruses. Two putative promoters, located upstream of the left and right ORFs, are presumed to regulate expression of NS and VP genes, respectively. Thus, IHHNV is closely related to densoviruses of the genus Brevidensovirus in the family Parvoviridae, and we therefore propose to rename this virus Penaeus stylirostris densovirus (PstDNV).

Amino Acid Sequence↗
PubMed2000

Transcriptional analysis of the ribonucleotide reductase genes of shrimp white spot syndrome virus.

The causative agent of white spot syndrome (WSS) is a large double-stranded DNA virus, WSSV, which is probably a representative of a new genus, provisionally called Whispovirus. From previously constructed WSSV genomic libraries of a Taiwan WSSV isolate, clones with open reading frames (ORFs) that encode proteins with significant homology to the class I ribonucleotide reductase large (RR1) and small (RR2) subunits were identified. WSSV rr1 and rr2 potentially encode 848 and 413 amino acids, respectively. RNA was isolated from WSSV-infected shrimp at different times after infection and Northern blot analysis with rr1- and rr2-specific riboprobes found major transcripts of 2.8 and 1.4 kb, respectively. 5' RACE showed that the major rr1 transcript started at a position of -84 (C) relative to the ATG translational start, while transcription of the rr2 gene started at nucleotide residue -68 (T). A consensus motif containing the transcriptional start sites for rr1 and rr2 was observed (TCAc/tTC). Northern blotting and RT-PCR showed that the transcription of rr1 and rr2 started 4-6 h after infection and continued for at least 60 h. The rr1 and rr2 genes thus appear to be WSSV "early genes."

Amino Acid Sequence↗
PubMed2000

Identification and characterization of a shrimp white spot syndrome virus (WSSV) gene that encodes a novel chimeric polypeptide of cellular-type thymidine kinase and thymidylate kinase.

From previously constructed genomic libraries of a Taiwan WSSV isolate, a putative WSSV tk-tmk gene was identified. Uniquely, the open reading frame (ORF) of this gene was predicted to encode a novel chimeric protein of 388 amino acids with significant homology to two proteins: thymidine kinase (TK) and thymidylate kinase (TMK). Northern blot analysis with a WSSV tk-tmk-specific riboprobe detected a major transcript of 1.6 kb. When healthy adult Penaeus monodon shrimp were inoculated with WSSV, the tk-tmk gene transcript was first detected by RT-PCR analysis at 4 h postinfection and transcription levels continued to increase over the first 18 h. The gene's major in vitro transcription and translation product, equivalent to the predicted size (43 kDa), is a single chimeric protein that includes both the TK and TMK functional motifs. Evidence for phylogenetic analysis and sequence alignment suggested that the gene may have resulted from the fusion of a cellular-type TK gene and a cellular-type TMK gene. Its unique arrangement may also provide a valuable gene marker for WSSV.

Amino Acid Sequence↗
PubMed2002

Identification of a nucleocapsid protein (VP35) gene of shrimp white spot syndrome virus and characterization of the motif important for targeting VP35 to the nuclei of transfected insect cells.

To identify the protein encoded by a 687-bp open reading frame (ORF) of a salI genomic DNA fragment of shrimp white spot syndrome virus (WSSV), we expressed the ORF in a baculovirus/insect cell expression system. The apparent molecular mass of the recombinant protein on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was 35 kDa in insect cells. Antibody raised against bacterially synthesized protein of the ORF identified a nucleocapsid protein (VP35) in the extracts of both the purified WSSV virions and the nucleocapsids which comigrated with the 35-kDa baculovirus-expressed recombinant protein on SDS-PAGE. We also show by transient expression in insect cells (Sf9) that VP35 targets the nucleus. Two potential nuclear localization signals (NLSs) were characterized, but only one of them was important for targeting VP35 to the nuclei of transfected insect cells. Replacement of a cluster of four positively charged residues ((24)KRKR(27)) at the N terminus of the protein with AAAA resulted in mutant proteins that were distributed only in the cytoplasm, thus confirming that this sequence is a critical part of the functionally active NLS of VP35.

Amino Acid Sequence↗