Demographic assessment of the blue crab (Callinectes sapidus) in Chesapeake Bay using extractable lipofuscins as age markers

The blue crab (Callinectes sapidus) plays an important economic and ecological role in estuaries and coastal habitats from the Gulf of Mexico to the east coast of North America, but demographic assessments are limited by length-based methods. We applied an alternative aging method using biochemical measures of metabolic byproducts (lipofuscins) sequestered in the neural tissue of eyestalks to examine population age structure. From Chesapeake Bay, subsamples of animals collected from the 1998–99 (n=769) and 1999–2000 (n=367) winter dredge surveys were collected and lipofuscin was measured. Modal analysis of the lipofuscin index provided separation into three modes, whereas carapace-width data collected among the same individuals showed two broad modes. Lipofuscin modal analysis indicated that most adults (carapace width >120 mm) were <2 years old. The results indicate that use of extractable lipofuscin can provide a more accurate and better resolved estimation of demographic structure of blue crab populations in the field than size alone.

Intertidal transect studies of northern Monterey Bay

In accordance with a contract dated 10/22/71 between the Association of the Monterey Bay Area Governments (AMBAG) and the University of California, Santa Cruz, (UCSC), two permanent intertidal transects with 14 permanent meter-square quadrats were established on the north shore of Monterey Bay during November, 1971. One transect (6 quadrats) was placed on the shore near the Santa Cruz Sanitation outfall, while the second (8 quadrats) was placed near the Eastcliff Sanitation District outfall at Soquel Polnt (Pleasure Point). Animals and plants within the quadrats were listed, their abundance estimated, and representative specimens collected for a reference collection maintained at UCSC. Additional species of animals and plants in the areas of the transects were collected for the reference collection. These collections will serve as a base-line for comparative studies which can follow the magnitude and direction of future changes in these areas.

On the behaviour of marine crustaceans in an electrical field of alternating current

The minimum flow of current essential to produce electro-narcosis and fixation in marine crustaceans, Metapenaeus affinis, Parapenaeopsis stylifera, Panulirus ornatus and Neptunus sanguinofentus in an AC field of specific strength varied with the lengths of animals tested, except in case of the last one where the increase in size of animals showed no significant effect on the effective period for different reactions. M. affinis in interrupted AC required threshold pulses for narcosis and fixation, which varied inversely with the length of the organism. Animals subjected to current of higher effective period required longer time for recovery in interrupted AC of 50 c/s. The period for narcosis and recovery of M. affinis did not vary significantly with repeated stimulations on the same organism.

Human activities and interests within and around lakes of the Victoria and Kyoga basin and their consequences to sustainable fish production

Of all the great lakes, Lake Victoria has the highest population concentration on its fringes. This has resulted into serious human impacts on the ecosystem through intense agricultural activities (cultivation, livestock and over fishing), sporadic settlements, urbanization and industrial establishments. The consequences have been loss of animals and plant life, deforestation and general land degradation, pollution, loss of water quality and clean air. Aquatic life has become endangered and less guaranteeing to continued fish production. Awareness workshops and general talks have been done to a few selected communities by the lakes landing sites and in the catchment area to mitigate the deteriorating environmental conditions. Naturally the situation calls for reversal to the increasing stress of the ecosystem. As a result, every water body surveyed put forward some mitigation suggestions

Comparison of stationary acoustic monitoring and visual observation of finless porpoises

The detection performance regarding stationary acoustic monitoring of Yangtze finless porpoises Neophocaena phocaenoides asiaeorientalis was compared to visual observations. Three stereo acoustic data loggers (A-tag) were placed at different locations near the confluence of Poyang Lake and the Yangtze River, China. The presence and number of porpoises were determined acoustically and visually during each 1-min time bin. On average, porpoises were acoustically detected 81.7 +/- 9.7% of the entire effective observation time, while the presence of animals was confirmed visually 12.7 +/- 11.0% of the entire time. Acoustic monitoring indicated areas of high and low porpoise densities that were consistent with visual observations. The direction of porpoise movement was monitored using stereo beams, which agreed with visual observations at all monitoring locations. Acoustic and visual methods could determine group sizes up to five and ten individuals, respectively. While the acoustic monitoring method had the advantage of high detection probability, it tended to underestimate group size due to the limited resolution of sound source bearing angles. The stationary acoustic monitoring method proved to be a practical and useful alternative to visual observations, especially in areas of low porpoise density for long-term monitoring.

Hopping robot based on free vibration of an elastic curved beam

This study presents a novel approach to the design of low-cost and energy-efficient hopping robots, which makes use of free vibration of an elastic curved beam. We found that a hopping robot could benefit from an elastic curved beam in many ways such as low manufacturing cost, light body weight and small energy dissipation in mechanical interactions. A challenging problem of this design strategy, however, lies in harnessing the mechanical dynamics of free vibration in the elastic curved beam: because the free vibration is the outcome of coupled mechanical dynamics between actuation and mechanical structures, it is not trivial to systematically design mechanical structures and control architectures for stable locomotion. From this perspective, this paper investigates a case study of simple hopping robot to identify the design principles of mechanics and control. We developed a hopping robot consisting of an elastic curved beam and a small rotating mass, which was then modeled and analyzed in simulation. The experimental results show that the robot is capable of exhibiting stable hopping gait patterns by using a small actuation with no sensory feedback owing to the intrinsic stability of coupled mechanical dynamics. Furthermore, an additional analysis shows that, by exploiting free vibration of the elastic curved beam, cost of transport of the proposed hopping locomotion can be in the same rage of animals' locomotion including human running. © 2011 IEEE.

Food habits of two-year-old Chinese mitten crab (Eriocheir sinensis) stocked in Lake Bao'an, China

Gastric mills of 362 specimens of two-year-old Chinese mitten crab (Eriocheir sinensis), which contained recognizable food items, from Lake Bao'an, China were examined. The food items were macrophytes, algae, arthropods, oligochaetes, fish, protozoa, rotifers, gastropods, and detritus, and the percent frequencies of occurrence (FO) for these items were 87.3%, 82.0%, 48.2%, 28.2%, 28.7%, 0.3%, 0.6%, 0.3% and 88.7%, respectively. Unidentified animal tissue was often observed and had a FO of 46.1%. In total, FO of plants (macrophytes + algae) was 87.7% and of animals was 89.8%. However, 5.8% of the gastric mills contained only animals, 5.3% had only macrophytes, and 0.3% contained only algae. There was no significant difference (p>0.05) in food habits between male and female crabs. The ratio of cell number of macrophytes to algae was about 156:1.

Characterization of degQ(Vh), a serine protease and a protective immunogen from a pathogenic Vibrio harveyi strain

Vibrio harveyi is an important marine pathogen that can infect a number of aquaculture species. V. harveyi degQ (degQ(Vh)), the gene encoding a DegQ homologue, was cloned from T4, a pathogenic V. harveyi strain isolated from diseased fish. DegQ(Vh) was closely related to the HtrA family members identified in other Vibrio species and could complement the temperature-sensitive phenotype of an Escherichia coli strain defective in degP. Expression of degQVh in T4 was modulated by temperature, possibly through the sigma(E)-like factor. Enzymatic analyses demonstrated that the recombinant DegQVh protein expressed in and purified from E. coli was an active serine protease whose activity required the integrity of the catalytic site and the PDZ domains. The optimal temperature and pH of the recombinant DegQVh protein were 50 C and pH 8.0. A vaccination study indicated that the purified recombinant DegQVh was a protective immunogen that could confer protection upon fish against infection by V. harveyi. In order to improve the efficiency of DegQVh as a vaccine, a genetic construct in the form of the plasmid pAQ1 was built, in which the DNA encoding the processed DegQVh protein was fused with the DNA encoding the secretion region of AgaV, an extracellular beta-agarase. The E.coli strain harboring pAQ1 could express and secrete the chimeric DegQVh protein into the culture supernatant. Vaccination of fish with viable E. coli expressing chimeric degQ(Vh) significantly (P < 0.001) enhanced the survival of fish against V. harveyi challenge, which was possibly due to the relatively prolonged exposure of the immune system to the recombinant antigen produced constitutively, albeit at a gradually decreasing level, by the carrier strain.

Characterization of degQ(Vh), a serine protease and a protective immunogen from a pathogenic Vibrio harveyi strain

Vibrio harveyi is an important marine pathogen that can infect a number of aquaculture species. V. harveyi degQ (degQ(Vh)), the gene encoding a DegQ homologue, was cloned from T4, a pathogenic V. harveyi strain isolated from diseased fish. DegQ(Vh) was closely related to the HtrA family members identified in other Vibrio species and could complement the temperature-sensitive phenotype of an Escherichia coli strain defective in degP. Expression of degQVh in T4 was modulated by temperature, possibly through the sigma(E)-like factor. Enzymatic analyses demonstrated that the recombinant DegQVh protein expressed in and purified from E. coli was an active serine protease whose activity required the integrity of the catalytic site and the PDZ domains. The optimal temperature and pH of the recombinant DegQVh protein were 50 C and pH 8.0. A vaccination study indicated that the purified recombinant DegQVh was a protective immunogen that could confer protection upon fish against infection by V. harveyi. In order to improve the efficiency of DegQVh as a vaccine, a genetic construct in the form of the plasmid pAQ1 was built, in which the DNA encoding the processed DegQVh protein was fused with the DNA encoding the secretion region of AgaV, an extracellular beta-agarase. The E.coli strain harboring pAQ1 could express and secrete the chimeric DegQVh protein into the culture supernatant. Vaccination of fish with viable E. coli expressing chimeric degQ(Vh) significantly (P < 0.001) enhanced the survival of fish against V. harveyi challenge, which was possibly due to the relatively prolonged exposure of the immune system to the recombinant antigen produced constitutively, albeit at a gradually decreasing level, by the carrier strain.

Using the ribosomal internal transcribed spacer (ITS) as a complement marker for species identification of red macroalgae

Barcodes based on mitochondrial cytochrome oxidase (mtDNA CO1) sequences are being used for broad taxonomic groups of animals with demonstrated success in species identification and cryptic species discovery, but it has become clear that complementation by a nuclear marker system is necessary, in particular for the barcoding of plants. Here, we propose the nuclear internal transcribed spacer (ITS) as a potentially usable and complementary marker for species identification of red macroalgae, as well as present a primary workflow for species barcoding. Data show that for most red macroalgal genera (except members of the family Delesseriaceae), the size of ITS region ranges from 600 to 1200 bp, and contains enough variation to generate unique identifiers at either the species or genus levels. Consistent with previous studies, we found that the ITS sequence can resolve closely related species with the same fidelity as mtDNA CO1. Significantly, we confirmed that length polymorphism in the ITS region (including 5.8S rRNA gene) can be utilized as a character to discriminate red macroalgal species. As a complementary marker, the verifiable nuclear ITS region can speed routine identification and the detection of species, advance ecological and taxonomic inquiry, and permit rapid and accurate analysis of red macroalgae.

Identification and immunoprotective analysis of an in vivo-induced Edwardsiella tarda antigen

Edwardsiella tarda is a severe aquaculture pathogen that can infect many important fish species cultured worldwide. The aim of this study was to evaluate the vaccine potential of an E. tarda antigen, Eta21, which was identified from a pathogenic E. tarda strain via the method of in vivo-induced antigen technology (IVIAT). Eta21 is 510-amino acid in length and shares similar to 58% sequence identity with a putative peptidase of several bacterial species. eta21 was subcloned into Escherichia colt, and recombinant Eta21 was purified as a histidine-tagged protein. When used as a subunit vaccine, purified recombinant Eta21 was effective against lethal E. tarda challenge in a Japanese flounder model. In order to improve the immunoprotective efficacy of Eta21, the chimera AgaV-Eta21 was constructed, which consists of Eta21 fused in-frame to the secretion domain of AgaV, an extracellular beta-agarase. E. coli DH5 alpha harboring plasmid pTAET21, which constitutively expresses agaV-eta21, was able to produce and secret AgaV-Eta21 into the extracellular milieu. Vaccination of Japanese flounder with live DH5 alpha/pTAET21 elicited immunoprotection that is significantly higher in level than that induced by vaccination with purified recombinant Eta21. Vaccination with DH5 alpha/pTAET21 and recombinant Eta21 both induced the production of specific serum antibodies at four to eight weeks post-vaccination. Taken together, these results demonstrate that Eta21, especially that delivered by DH5 alpha/pTAET21, is an effective vaccine candidate against E. tarda infection. (C) 2009 Elsevier Ltd. All rights reserved.

Immunoprotective analysis of VhhP2, a Vibrio harveyi vaccine candidate

VhhP2 is an Outer membrane protein identified in a pathogenic Vibrio harveyi strain, T4, isolated from diseased fish. When used as a Subunit Vaccine, purified recombinant VhhP2 affords high level of protection upon Japanese flounder against V harveyi challenge. Vaccination with VhhP2 induced the expression of a number of immune-related genes, especially those encoding immunoglobulin M (IgM) and major histocompatibility complex (MHC) II alpha. A VhhP2 surface display system, in the form of the fish commensal strain FIR harboring the vhhP2-expressing plasmid pJVP, was constructed. PF3/pJVP is able to produce and present recombinant VhhP2 on cell surface. Vaccination of fish with live PF3/pJVP via intraperitoneal injection elicited Strong immunoprotection. Vaccination of fish orally with live PF3/pJVP embedded in alginate microspheres also induced effective immunoprotection. In addition, a VhhP2-based surface display system was created, in which VhhP2 serves as a carrier for the Surface delivery of a heterologous Edwardsiella tarda immunogen, Et18, that is fused in-frame to VhhP2. DH5 alpha/pJVP18, which expresses and surface-displays the VhhP2-Et18 chimera, proved to be an effective vaccine that call protect fish against infections by V. harveyi and E. tarda to the extents comparable to those produced by vaccination with purified recombinant VhhP2 and Et18, respectively. These data suggest that VhhP2 may be applied as a vaccine and a vaccine carrier against infections by V. harveyi and other pathogens such as F. tarda. (C) 2009 Elsevier Ltd. All rights reserved.

Construction of an attenuated Pseudomonas fluorescens strain and evaluation of its potential as a cross-protective vaccine

Ferric uptake regulator (Fur) is a global transcription regulator that is ubiquitous to Gram-negative bacteria and regulates diverse biological processes, including iron uptake, cellular metabolism, stress response, and production of virulence determinants. As a result, for many pathogenic bacteria, Fur plays a crucial role in the course of infection and disease development. In this study, the fur gene was cloned from a pathogenic Pseudomonas fluorescens strain, TSS, isolated from diseased Japanese flounder cultured in a local farm. TSS Fur can partially complement the defective phenotype of an Escherichia coli fur mutant. A TSS fur null mutant, TFM, was constructed. Compared to TSS, TFM exhibits reduced growth ability, aberrant production of outer membrane proteins, decreased resistance against host serum bactericidal activity, impaired ability to disseminate in host blood and tissues, and drastic attenuation in overall bacterial virulence in a Japanese flounder infection model. When used as a live vaccine administered via the injection, immersion, and oral routes, TFM affords high levels of protection upon Japanese flounder against not only P.fluorescens infection but also Aeromonas hydrophila infection. Furthermore, a plasmid, pJAQ, was constructed, which expresses the coding element of the Vibrio harveyi antigen AgaV-DegQ. TFM harboring pJAQ can secret AgaV-DegQ into the extracellular milieu. Vaccination of Japanese flounder with live TFM/pJAQ elicited strong immunoprotection against both V. harveyi and A. hydrophila infections. (C) 2009 Elsevier Ltd. All rights reserved.

Expression profiles of penaeidin from Fenneropenaeus chinensis in response to WSSV and vibrio infection by real-time PCR

Penaeidin from Chinese shrimp (Fenneropenaeus chinensis) has proved to be one of the most important antimicrobial peptides in the bodies of animals. The relative quantitative real-time PCR method is developed to study through time, the mRNA expression profile of penaeidin in the muscle and haemocyte tissue of Chinese shrimp infected with vibrio (Vibrio anguillarum) and WSSV (white spot syndrome virus). Research results showed that the same pathogens infection experiments produced similar gene expression profile in different tissues while different expression profiles appeared in the same tissues infected by different exterior pathogens. In vibrio infection experiments, a "U" Re expression profile resulted. Expression levels of penaeidin increased and surpassed the non-stimulated level, indicating that penaeidin from Chinese shrimp has noticeable antimicrobial activities. In WSSV infection experiments, the expression profile appeared as an inverse "U" with the expression of penaeidin gradually decreasing to below baseline level after 24 h. The expression of antimicrobial peptides gene in mRNA level in response to virus infection in shrimp showed that international mechanisms of virus to haemocytes and microbial to haemocytes are completely different. Decline of penaeidins expression levels may be due to haemocytes being destroyed by WSSV or that the virus can inhibit the expression of penaeidins by yet undiscovered modes. The expression profiles of penaeidin in response to exterior pathogen and the difference of expression profiles between vibrio and WSSV infection provided some clues to further understanding the complex innate immune mechanism in shrimp.

Expression of Scophthalmus maximus CD83 correlates with bacterial infection and antigen stimulation

CD83 is a transmembrane glycoprotein of the immunoglobulin (Ig) superfamily and a surface marker for fully matured dendritic cells (DCs) in humans and mice. In teleosts, DC-like cells and their molecular markers are largely unknown. In this report, we described the identification and expressional analysis of a CD83 homologue, SmCD83, from turbot Scophthalmus maximus. The open reading frame of SmCD83 is 639 bp, which is preceded by a S'-untranslated region (UTR) of 87 bp and followed by a 3'-UTR of 1111 bp. The SmCD83 gene is 4716 bp in length, which contains five exons and four introns. The deduced amino acid sequence of SmCD83 shares 40-50% overall identities with the CD83 of several fish species. Like typical CD83, SmCD83 possesses an Ig-like extracellular domain, a transmembrane domain, and a cytoplasmic domain. The conserved disulfide bond-forming cysteine residues and the N-linked glycosylation sites that are preserved in CD83 are also found in SmCD83. Expressional analysis showed that constitutive expression of SmCD83 was high in gill, blood, spleen, muscle, and kidney and low in heart and liver. Bacterial infection and poly(I:C) treatment enhanced SmCD83 expression in kidney in time-dependent manners. Likewise, bacterial challenge caused significant induction of SmCD83 expression in cultured macrophages. Vaccination of turbot with a bacterin and a purified recombinant subunit vaccine-induced significant SmCD83 expression during the first week following vaccination. These results demonstrate that SmCD83 expression correlates with microbial challenge and antigen stimulation, which suggests the possibility that there may exist in turbot DC-like antigen-presenting cells that express SmCD83 upon activation by antigen uptake. In addition, these results also suggest that SmCD83 may serve as a marker for activated macrophages in turbot. (C) 2010 Elsevier Ltd. All rights reserved.