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At least 883 records · Page 49Linked to original sources

Molecule capture by olfactory antennules: mantis shrimp.

A critical step in the process of olfaction is the movement of odorant molecules from the environment to the surface of a chemosensory structure. Many marine crustaceans capture odorant molecules with arrays of chemosensory sensilla (aesthetascs) on antennules that they flick through the water. We developed a model to calculate molecule flux to the surfaces of aesthetascs in order to study how the size, aesthetasc spacing, and flick kinematics of olfactory antennules affect their performance in capturing molecules from the surrounding water. Since the three-dimensional geometry of an aesthetasc-bearing antennule is complex, dynamically-scaled physical models can often provide an efficient method of determining the fluid velocity field through the array. Here we present a method to optimize the incorporation of such measured velocity vector fields into a numerical simulation of the advection and diffusion of odorants to aesthetasc surfaces. Furthermore, unlike earlier models of odorant interception by antennae, our model incorporates odorant concentration distributions that have been measured in turbulent ambient flows. By applying our model to the example of the olfactory antennules of mantis shrimp, we learned that flicking velocity can have profound effects on odorant flux to the aesthetascs if they operate in the speed range in which the leakiness of the gaps between the aesthetascs to fluid movement is sensitive to velocity. This sensitivity creates an asymmetry in molecule fluxes between outstroke and return stroke, which results in an antennule taking discrete samples in space and time, i.e. "sniffing". As stomatopods grow and their aesthetasc Reynolds number increases, the aesthetasc arrangement on the antennule changes in a way that maintains these asymmetries in leakiness and molecule flux between the outstroke and return stroke, allowing the individual to continue to take discrete samples as it develops.

Animals↗

Tuning of photoreceptor function in three mantis shrimp species that inhabit a range of depths. II. Filter pigments.

Within single species of stomatopod crustaceans, visual pigment classes of homologous photoreceptors throughout the retina are identical in all individuals and do not vary with the spectral characteristics of local habitats. We examined whether spectral sensitivities of stomatopod photoreceptors are differentially tuned through variations in the filter pigments associated with particular receptor classes. All classes of intrarhabdomal filters were characterized using microspectrophotometry in retinas of three stomatopod species, Haptosquilla trispinosa, Gonodactylellus affinis, and Gonodactylopsis spongicola, comparing individuals of each species collected from shallow or deep water. Depending on the depth of collection, filters varied among individuals both in optical density and in spectral shape, and the variation that was observed was similar in all three species. The changes in filter density and spectrum increased absolute sensitivity in retinas of animals living at greater depths, and tuned their long-wavelength photoreceptors for improved function in the bluer light available in deep water. Plasticity in retinal spectral function may be common in mantis shrimp species that occupy a range of habitat depths.

Animals↗

Transcriptional profile of shrimp white spot syndrome virus (WSSV) genes with DNA microarray.

In an attempt to obtain the transcriptional profile of shrimp white spot syndrome virus (WSSV) genes, DNA microarray analysis was performed using amplified DNA fragments of the WSSV genome that covered most of the presumptive open reading frames (ORFs). Total RNAs were extracted from WSSV-infected crayfish (Cambarus clarkii) and reverse transcribed into cDNA and labeled with 32P-dATP. The DNA microarray detected 81.1% of the ORFs, and 47 of these were transcribed at 6 h post infection, suggesting they may be early genes. Some early genes were further confirmed by temporal analyses of gene transcription. Results in this study indicated the effectiveness of DNA microarray for discovering early viral genes.

Animals↗

Protein analysis of geographic isolates of shrimp white spot syndrome virus.

Six geographic isolates of the white spot syndrome virus (WSSV) of penaeid shrimp, from China, India, Thailand, South Carolina, Texas, as well as from crayfish kept at the US National Zoo in Washington D. C, were compared by electron microscopy and sodium sulfate polyacrylamine gel electrophoresis (SDS-PAGE). Amino acid compositions of four of the major structural polypeptides of the South Carolina WSSV were analyzed, and three of the four polypeptides were partially sequenced from their NH2 termini. The morphologies of purified virions of the six geographic isolates of WSSV were indistinguishable by transmission electron microscopy. By SDS-PAGE, the protein profiles of the six isolates were very similar, but not identical. They all contained three major polypeptides with sizes of approximately 25, 23 and 19 kDa. A fourth major polypeptide at the 14.5 kDa position was observed in four of the geographic isolates. The WSSV isolated from crayfish presented a slightly different structural protein profile, particularly with regard to the protein in the 19 kDa range that appeared larger in size than those of the other isolates. The NH2 terminal amino acids of the 25, 23 and 14.5 kDa polypeptides of the South Carolina WSSV were sequenced as MDLSFTLSVVTA, MEFGNLTNLDVA, and VARGGKTKGRRG, respectively. No significant homologous sequence was found in the GenBank. These protein sequences have been submitted to the SWISS-PROT Protein Data Bank and assigned the accession numbers P82004, P82005 and P82006.

Amino Acid Sequence↗

Genetic heterogeneity of the giant tiger shrimp (Penaeus monodon) in Thailand revealed by RAPD and mitochondrial DNA RFLP analyses.

Genetic diversity of the giant tiger shrimp (Penaeus monodon) collected from 5 areas, Chumphon and Trat (Gulf of Thailand), and Phangnga, Satun, and Trang (Andaman Sea), was examined by randomly amplified polymorphic DNA (RAPD) and mitochondrial DNA (16S ribosomal DNA and an intergenic COI-COII) polymorphism. A total of 53 polymorphic fragments from UBC299, UBC273, and UBC268 was consistently scored across all samples. From the respective primers 26, 32, and 30 genotypes were generated. A 260-bp RAPD fragment generated by the primer UBC268 was specifically observed in 95.8% of Trat P. monodon, suggesting that this RAPD could be used as a marker for comparing phenotypic performance of P. monodon from Trat and other geographic samples. In addition, 37 mtDNA composite haplotypes were observed from restriction analysis of the same P. monodon samples. High haplotype diversity (0.855) and nucleotide diversity (3.328%) of Thai P. monodon were observed. Population differentiation of P. monodon between the Andaman Sea and Gulf of Thailand was clearly illustrated by both techniques (P <.0001). Nevertheless, contradictory results on patterns of differentiation were observed between P. monodon within the Gulf of Thailand. Analysis of nuclear DNA polymorphism (RAPD) indicated a genetically significant difference between Chumphon and Trat ( P <.0001), whereas mtDNA polymorphism did not show differentiation between these samples (P =.0497). Under the presumption of selective neutrality of these markers, biased female gene flow between Trat and Chumphon P. monodon may exist and be responsible for an anomalous differentiation pattern between these geographic samples.

Journal Article↗

Crustins, homologues of an 11.5-kDa antibacterial peptide, from two species of penaeid shrimp, Litopenaeus vannamei and Litopenaeus setiferus.

The response of crustaceans to pathogens is believed to depend solely on innate, nonadaptive immune mechanisms, including phagocytosis, encapsulation, clotting, and a variety of soluble antimicrobial activities. Arthropod antimicrobial peptides, while characterized primarily from insects, also have been isolated from crustaceans. Expressed sequence tag analysis of hemocyte complementary DNA libraries from 2 species of shrimp, Litopenaeus vannamei and Litopenaeus setiferus, revealed transcripts with strong sequence similarity to an 11.5-kDa antibacterial peptide (crustin Cm1) found in Carcinus maenas. Crustins were also observed to contain motifs common to proteinase inhibitors. Analysis of these cDNA libraries yielded at least 3 different isoforms of this peptide in L. vannamei (crustin Lv1-Lv3) and 3 in L. setiferus (crustin Ls1-Ls3). Further analysis of a second L. vannamei cDNA library revealed the presence of 3 more possible isoforms (crustin Lv4-Lv6), which differed from those seen in the first L. vannamei cDNA library. Genomic Southern blot analysis revealed a complex family of crustin-related sequences. However, full-length crustin appears to be encoded by a much more restricted subset of sequences within this family.

Journal Article↗

Identification of immune-related genes in hemocytes of black tiger shrimp (Penaeus monodon).

An expressed sequence tag (EST) library was constructed from hemocytes of the black tiger shrimp (Penaeus monodon) to identify genes associated with immunity in this economically important species. The number of complementary DNA clones in the constructed library was approximately 4 x 10(5). Of these, 615 clones having inserts larger than 500 bp were unidirectionally sequenced and analyzed by homology searches against data in GenBank. Significant homology to known genes was found in 314 (51%) of the 615 clones, but the remaining 301 sequences (49%) did not match any sequence in GenBank. Approximately 35% of the matched ESTs were significantly identified by the BLASTN and BLASTX programs, while 65% were recognized only by the BLASTX program. Of the 615 clones, 55 (8.9%) were identified as putative immune-related genes. The isolated genes were composed of those coding for enzymes and proteins in the clotting system and the prophenoloxidase-activating system, antioxidative enzymes, antimicrobial peptides, and serine proteinase inhibitors. Three full-length ESTs encoding antimicrobial peptides (antilipopolysaccharide and penaeidin homologues) and a heat shock protein (cpn10 homologue) are reported.

Journal Article↗

High frequency and large number of polymorphic microsatellites in cultured shrimp, Penaeus (Litopenaeus) vannamei [Crustacea:Decapoda].

A total of 1479 recombinant clones were obtained from a Sau3A-digested genomic library of Penaeus (Litopenaeus) vannamei and used for probe hybridization. Of the 251 clones that tested positive to one or more of the probes and were sequenced, 173 (69%) contained 573 simple sequence repeats, or microsatellites, with 3 or more repeats. The frequency of microsatellites with 3, 5, and 10 or more repeats was 1 in 0.94 kb, 1 in 2.78 kb, and 1 in 5.94 kb, respectively. To increase the number of polymorphic markers for mapping, 136 primer sets that flanked microsatellites containing single or multiple motifs with 3 or more repeats were designed and tested. Of the 136 primers, 93 (68.0%) were polymorphic in cultured shrimp, with polymorphism information content (PIC) values ranging from 0.195 to 0.873, and observed heterozygosities ranging from 10% to 100%. These markers are being used along with other markers to construct a linkage map for P. vannamei.

Animals↗

Genetic variability assessed by microsatellites in a breeding program of Pacific white shrimp (Litopenaeus vannamei).

Genetic diversity in a shrimp-breeding program was monitored for 2 generations by microsatellite DNA markers (Pvan1578 and Pvan1815) to establish levels of variation and proceed with a selection program. An increase in the number and frequencies of some alleles in both microsatellite loci from G0 to G2 was induced by foreign sire contributions. Most common alleles and high heterozygosities (around 70% in both loci) were maintained through the generations, indicating that there had not been a significant loss of genetic variability in the breeding program. However, when compared with variability in other wild and cultured stocks, the presence of 4 main alleles at both loci may be an indication that a certain reduction in variability already was present in the line used as founder stock (G0). Therefore, it is recommended that additional genetic variability be introduced to the breeding stock by crossing it with a different line.

Animals↗

Recombinant expression and characterization of five-domain Kazal-type serine proteinase inhibitor of black tiger shrimp (Penaeus monodon).

Four full-length complementary DNAs of Kazal-Type serine proteinase inhibitors (SPIs) were identified from hemocyte cDNA libraries of the black tiger shrimp Penaeus monodon and recognized as having 4 (SPIPm1) and 5 (SPIPm2, SPIPm3, and SPIPm4) Kazal domains. SPIPm2 encoding a complete 5-Kazal-domain inhibitor was expressed in the Escherichia coli system. Inhibitory activity of crude proteins against various serine proteases was tested using SPI activity gelatin with sodium dodecylsulfate polyacrylamide gel electrophoresis and inhibitory spectrum assays. A 32-kDa recombinant protein (rSPIPm2) showed inhibitory activity against trypsin, chymotrypsin, and subtilisin, but not elastase. Concordantly, inhibitory spectrum assays showed that crude rSPIPm2 strongly inhibited trypsin (89%) and chymotrypsin (70%), but less effectively inhibited subtilisin (8%), and did not inhibit elastase activity. Northern blot analysis of hemocyte total RNA showed 2 SPI transcripts of 1.6 and 1.7 kb in size. Tissue-specific expression using reverse transcriptase polymerase chain reaction suggests that SPIPm2 is exclusively expressed in hemocytes of P. monodon.

Amino Acid Sequence↗

Development of EST-SSR markers by data mining in three species of shrimp: Litopenaeus vannamei, Litopenaeus stylirostris, and Trachypenaeus birdy.

We report on the data mining of publicly available Litopenaeus vannamei expressed sequence tags (ESTs) to generate simple sequence repeat (SSRs) markers and on their transferability between related Penaeid shrimp species. Repeat motifs were found in 3.8% of the evaluated ESTs at a frequency of one repeat every 7.8 kb of sequence data. A total of 206 primer pairs were designed, and 112 loci were amplified with the highest success in L. vannamei. A high percentage (69%) of EST-SSRs were transferable within the genus Litopenaeus. More than half of the amplified products were polymorphic in a small testing panel of L. vannamei. Evaluation of those primers in a larger testing panel showed that 72% of the markers fit Hardy-Weinberg equilibrium, which shows their utility for population genetic analysis. Additionally, a set of 26 of the EST-SSRs were evaluated for Mendelian segregation. A high percentage of monomorphic markers (46%) proved to be polymorphic by singles-stranded conformational polymorphism analysis. Because of the high number of ESTs available in public databases, a data mining approach similar to the one outlined here might yield high numbers of SSR markers in many animal taxa.

Animals↗

Synthesis of Hemolymph High-Density Lipoprotein beta-Glucan Binding Protein by Penaeus vannamei Shrimp Hepatopancreas.

It was recently shown that the shrimp high-density lipoprotein (HDL) and the beta-1,3-glucan binding protein (BGBP) are identical, implying dual functions for the same protein: lipid transport and involvement in the defense system. Because this protein is present in plasma, and the hepatopancreas is a major lipid storage gland, we investigated the presence of the HDL/BGBP polypeptide and its messenger RNA in this tissue using a monospecific antibody against HDL/BGBP. Hepatopancreas crude protein extracts, as well as polypeptides produced by poly(A)(+) RNA in vitro translation, were recognized by the anti-HDL/BGBP. Furthermore, a specific pattern was revealed in hepatopancreas thin sections by immunodetection. Strong recognition was seen in the epithelial cells of hepatopancreatic tubules, probably related to the secretion process of this protein.

Journal Article↗

Genetic diversity of the giant tiger shrimp (Penaeus monodon) in Thailand revealed by PCR-SSCP of polymorphic EST-derived markers.

A total of 90 ESTs from normal and 157 from subtractive ovarian cDNA libraries of the giant tiger shrimp (Penaeus monodon) were sequenced. SSCP analysis of disulfide isomerase (DSl), zinc finger protein (ZFP), PMO920, and PMT1700 was carried out for population genetic studies of P. monodon in Thai waters. The number of codominant alleles per locus for overall samples was 6 for PMO920, 5 for PMT1700, and 12 for ZFP, and there were 19 dominant alleles for DSI. The observed heterozygosity of each geographic sample was 0.3043-0.5128 for PMO920, 0.3462-0.4643 for PMT1700, and 0.5000-0.8108 for ZFP. Linkage disequilibrium analysis indicated that genotypes of these loci segregate randomly (P > 0.05). Low genetic distance was found between pairs of geographic samples (0.0077-0.0178). The neighbor-joining tree constructed from the average genetic distance of overall loci allocated the Andaman samples (Satun, Trang, and Phangnga) into one cluster, and Chumphon and Trat into other clusters. Geographic differentiation between Satun-Trat and Satun-Phangnga was found only at the ZFP locus (P < 0.05), suggesting low degrees of genetic subdivision of Thai P. monodon.

Alleles↗

A genetic linkage map of Pacific white shrimp (Litopenaeus vannamei): sex-linked microsatellite markers and high recombination rates.

Pacific white shrimp (Litopenaeus vannamei) is the leading species farmed in the Western Hemisphere and an economically important aquaculture species in China. In this project, a genetic linkage map was constructed using amplified fragment length polymorphism (AFLP) and microsatellite markers. One hundred and eight select AFLP primer combinations and 30 polymorphic microsatellite markers produced 2071 markers that were polymorphic in either of the parents and segregated in the progeny. Of these segregating markers, 319 were mapped to 45 linkage groups of the female framework map, covering a total of 4134.4 cM; and 267 markers were assigned to 45 linkage groups of the male map, covering a total of 3220.9 cM. High recombination rates were found in both parental maps. A sex-linked microsatellite marker was mapped on the female map with 6.6 cM to sex and a LOD of 17.8, two other microsatellite markers were also linked with both 8.6 cM to sex and LOD score of 14.3 and 16.4. The genetic maps presented here will serve as a basis for the construction of a high-resolution genetic map, quantitative trait loci (QTLs) detection, marker-assisted selection (MAS) and comparative genome mapping.

Animals↗

Characterization of an early gene (wsv477) from shrimp white spot syndrome virus (WSSV).

White spot syndrome virus (WSSV) is one of the most virulent pathogens causing high mortality in shrimp. The viral early genes play a key role in DNA replication and virus proliferation. In this study, a WSSV gene (wsv477) encoding 208 amino acid peptides was characterized as an early gene. The temporal analysis showed that the wsv477 gene was first transcribed at 4 h post-infection, suggesting that it was an early gene. The wsv477 gene was expressed in Escherichia coli and purified. Subsequently the specific antibody was raised using the purified fusion protein (GST-WSV477). Western blot revealed that the wsv477 gene was expressed at 6 h post-infection in vivo. As indicated by GTP-binding assay, the WSV477 protein had GTP-binding activity.

Amino Acid Sequence↗

Lipid and fatty acid composition and energy partitioning during embryo development in the shrimp Macrobrachium borellii.

Energy partitioning, composition of lipids and fatty acids, and their utilization by embryos were determined in the lecithotrophic shrimp Macrobrachium borellii during seven development stages. The biochemical composition at stage I is represented by lipids, proteins, and carbohydrates, with 29.3, 28.7, and 0.2% dry weight, respectively. The former two were identified as the major energy-providing components, contributing 131 and 60 cal/100 mg egg, dry weight, respectively. The overall conversion efficiency (CE) was 45.0% (calculated as percentage of vitelline energy transformed into embryonic tissues). Lipids were the most important energy reserve (CE 39.3%), followed by proteins (CE 57.1%), both being simultaneously utilized during development while carbohydrates were synthesized de novo (CE 587.5%). Variation in the lipid class composition of embryos and vitellus showed an accumulation of triacylglycerols (TAG) and phospholipids (PL) up to stage IV, a more active accumulation and selective utilization phase (stages V and VI), and a consumption and de novo synthesis period until hatching. Structural lipids (PL and cholesterol) and pigment astaxanthin were selectively conserved in embryos, but TAG, hydrocarbons, and esterified sterols were preferentially depleted. Monounsaturated fatty acids (FA) were the major group in TAG, whereas polyunsaturated FA (PUFA) were the major group in PL after organogenesis. Certain PUFA such as 22:6n-3 and 20:5n-3 were selectively accumulated in PL.

Animals↗

Changes in phosphatidylcholine molecular species in the shrimp Macrobrachium borellii in response to a water-soluble fraction of petroleum.

The effect of the water-soluble fraction (WSF) of crude oil on lipid contents, lipid classes, FA, and PC molecular species was studied in high-phospholipid (hepatopancreas) and low-phospholipid (egg) tissues of a freshwater crustacean. After a 21-d exposure to a sublethal concentration of WSF, a significant decrease in shrimp total lipids was observed, although no alterations could be detected in the hepatopancreas or egg lipid contents. TAG/phospholipid ratios increased in the hepatopancreas and decreased in the eggs, suggesting alterations either in the mobilization of TAG to phospholipid pools or in the energy balance. The FA composition of phosphoglycerides in the hepatopancreas and eggs was dominated by PUFA, whereas the n-3/n-6 ratio was not affected by WSF exposure, although there was a significant increase in hepatopancreas 18:1n-9. Analysis of the PC molecular species by HPLC-ELSD showed the presence of 15 species, with 16:0/18:1, 18:1/18:2, 16:0/20:5, and 16:1/20:5 being the major species in the hepatopancreas. The PC molecular species in the eggs showed a different pattern, dominated by 16:0/18:1 and 18:1/18:2. Of the PC molecular species, 10 contained 22:6n-3, 20:5n-3, and 20:4n-6. Small amounts of di-PUFA species were also found. Exposure to WSF altered the PC molecular species in both tissues. The four major hepatopancreas molecular species and most of the ones containing PUFA decreased. This was compensated for by an increase in 16:1/18:1 (152%) and 18:1/18:1 (50%). The two major egg PC molecular species decreased, whereas the PUFA-containing ones increased. The contrasting responses of both tissues to WSF contamination suggests the presence of different homeostatic mechanisms.

Animals↗

The amino acid sequence of the fluorescein-labeled peptides of electric ray and brine shrimp (Na,K)-ATPase.

(Na,K)-ATPase from Torpedo californica (electric ray) and Artemia salina (brine shrimp) was labeled with fluorescein 5'-isothiocyanate (FITC) with concomitant loss of activity. Both inactivation and binding were inhibited in the presence of ATP. The sequence of the peptide resulting from tryptic digest containing labeled lysine from both enzymes is Tyr-Leu-Leu-Val-Met-Lys*-Gly-Ala-Pro-Glu-Arg. Thus the primary structure of this region is shown to be conserved in the enzymes of a nonvertebrate and a vertebrate.

Amino Acid Sequence↗