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PubMed2002

Expression and distribution of penaeidin antimicrobial peptides are regulated by haemocyte reactions in microbial challenged shrimp.

Penaeidins are a family of antimicrobial peptides constitutively produced and stored in the haemocytes of penaeid shrimp. In response to microbial stimulation, they are released into the blood circulation and they further attach to shrimp cuticle surfaces through a chitin-binding property. In the present paper, we have analysed their expression, regulation and distribution in shrimp tissues in response to experimental microbial challenge. We have shown that penaeidin mRNA and protein are restricted to granular haemocytes and that their expression and distribution are regulated through dramatic changes in haemocyte populations, both circulating and infiltrating shrimp tissues. Two distinct phases in the immune reactions were evidenced: (a) a migration of haemocytes towards the infection site within the first 12 h following microbial injection, with a local and massive release of peptides; (b) the appearance into the blood circulation and tissues of a haemocyte population displaying increased penaeidin-transcriptional activity, which may correspond to a systemic reaction involving haemocyte proliferation process. Finally, in vitro confrontation of haemocytes and bacteria revealed that penaeidins are released from granular haemocytes by a novel phenomenon of intracellular degranulation, probably followed by the lysis of the cells. Furthermore, penaeidins were shown covering bacterial surfaces suggesting that the peptides could be involved in opsonic activity. Penaeidin-positive bacteria were observed to be phagocytosed mainly by hyaline cells, a population that does not express penaeidins.

Animals↗

PubMed1999

Evaluation of toxicity of some saponins on brine shrimp.

Toxicity of several types of saponins (1-11) against brine shrimp (Artemia salina) were evaluated. As a result, it was found that most tested compounds were not toxic to brine shrimp at high enough concentration. The most toxic saponin (1) to brine shrimp showed also cytotoxicity towards HL-60 tumor cell line using MTT assay. Brine shrimp model may thus be used as bench-top assay in finding cytotoxic components from saponin-containing fractions of plant extracts.

Animals↗

PubMed1984

Toxicity of a phthalate ester in the diet of a penaied shrimp.

The toxicity of di-2-ethylhexyl phthalate (DEHP), a ubiquitous environmental pollutant, was measured experimentally as a contaminant in shrimp aquaculture feeds. Diets containing 40 to 50,000 ppm DEHP were fed to Penaeus vannamei for 14 d at 4% body weight/d. DEHP concentrations in shrimp, diet, and water were measured by electron-capture gas chromatography. Whole-body residues in shrimp were 18 ppm at the highest dose, and bioconcentration factors were inversely proportional to dose. DEHP in water was less than or equal to 1.7 ppb for all dose levels. No increased mortality or histopathological alterations were observed at any dose. Absorption of DEHP by P. vannamei was measured in static 24- and 96-h bioassays. Diets enriched with [14C]DEHP to levels of 60, 600, and 6000 ppm were fed at 2% body weight/d. At all dose levels, 3.7% of total radioactivity was measured as body burden in shrimp and 40% as polar species in test water after 96 h. DEHP by oral administration was absorbed, metabolized, and excreted, and this process was linear with dose for the dose range studied.

Analysis of Variance↗

PubMed2003

Lethal and sub-lethal effects of the fungicide chlorothalonil on three life stages of the grass shrimp, Palaemonetes pugio.

Chlorothalonil (2,4,5,6-tetrachloroisophthalonitrile) is the second most widely used fungicide in the United States. Due to the widespread use of chlorothalonil, it is important to investigate the effects chlorothalonil may have on estuarine species such as the grass shrimp, Palaemonetes pugio. This study examined the toxicity of chlorothalonil to three life-history stages (embryo, larvae, adult) of the grass shrimp. Also, molting frequency, growth response and metamorphosis from a larval life cycle pulsed exposure assay were examined as sub-lethal indicators of chlorothalonil exposure. Results showed embryos were the least sensitive with a 96-h Median Lethal Concentration (LC50) of 396.0 microg/L (95% Confidence Interval [CI] 331.3-472.4 microg/L). The adult 96-h LC50 was 152.9 microg/L (95% CI 120.3-194.5 microg/L). Larvae were the most sensitive to chlorothalonil exposure with a 96-h LC50 of 49.5 microg/L (95% CI 44.4-55.27 microg/L). In the life cycle pulsed exposure assay, all surviving larvae in the treatments required significantly more molts to reach postlarvae than the control. Other measured parameters showed differences between treatments and control but there was no statistical significance. This research demonstrated that chlorothalonil is highly toxic to grass shrimp and that larval grass shrimp would be the most appropriate life stage to use for chlorothalonil risk assessments since that stage is the most sensitive.

Animals↗

PubMed1984

Amino acid sequence of alpha chain of sarcoplasmic calcium binding protein obtained from shrimp tail muscle.

The isotypes of sarcoplasmic Ca2+ binding protein (SCP) were purified from shrimp tail muscle. SCP exists in a dimeric form. One sample of shrimp contained only alpha A chain, whereas another contained alpha B and beta chains, and a heterodimer of alpha B beta which was not analyzed precisely. The amino acid sequences of the two alpha chains were determined. The two alpha chains are composed of 190 and 192 amino acid residues, respectively. The sequences of the two alpha chains differed in only four amino acids out of 192 residues. The sequences indicate that the alpha chain has three Ca2+-binding sites which are common to EF-hand type Ca2+-binding protein. In the absence of added Ca2+ and Mg2+, the amounts of bound Ca2+ in alpha A, alpha B, and beta chains were 3.0, 3.3, and 2.4 mol/22,000 g protein, respectively. Thus, it is suggested that all three isotypes of shrimp SCP have three Ca2+-binding sites which have high affinity to Ca2+. The sequence homology of shrimp SCP with other EF-hand type Ca2+-binding proteins is very low. The protein having the greatest homology with this SCP was cod parvalbumin; the sequence homology is 18%.

Amino Acid Sequence↗

PubMed1984

Amino acid sequence of the beta chain of sarcoplasmic calcium binding protein (SCP) obtained from shrimp tail muscle.

The amino acid sequence of the beta chain of shrimp SCP has been determined. It is composed of 192 amino acid residues and is acetylated at the N-terminus. The molecular weight was determined to be 21,960. The sequence difference with the aB chain of the same shrimp, of which the sequence was determined previously (J Biochem. (1984) 95, 1603-1615), is 19% (37 non-identical residues out of 192 residues). The shrimp SCPs have three EF-hand type Ca2+ binding sites, however, from comparison with the amino acid sequences of SCPs of scallop (Takagi et al., Biochim. Biophys. Acta in press) and of sandworm (Kobayashi et al., manuscript in preparation), it is reasonable to think that SCP originally had four Ca2+ binding sites, and in the case of shrimp SCPs, one of them (site IV) may have lost the affinity to Ca2+ on amino acid replacements during evolution.

Amino Acid Sequence↗

PubMed1993

Partial cloning of the genome of infectious hypodermal and haematopoietic necrosis virus, an unusual parvovirus pathogenic for penaeid shrimps; diagnosis of the disease using a specific probe.

The infectious hypodermal and haematopoietic necrosis virus (IHHNV), pathogenic for penaeid shrimp, is an icosahedral unenveloped particle, 22 nm in diameter, with an ssDNA linear genome, and proposed to be a member of the Parvoviridae. A large majority of minus-strand DNA is incorporated into the capsids compared to the plus-strand. A small amount of reannealed plus- and minus-strands (dsDNA) obtained after nucleic acid extraction was blunt-ended and cloned into the system pUC18/Escherichia coli strain DH5 alpha. Selected clones were studied and characterized using restriction enzymes. One of them, BQ31, was used to construct different sized probes labelled with digoxigenin-11-dUTP. These probes failed to hybridize with DNA of some insect parvoviruses and with DNA of a parvo-like virus of shrimp. They reacted strongly with dilutions of homogenized IHHNV-infected shrimp tissues and, conversely, did not react with uninfected shrimp tissues. They hybridized in situ, in sections of infected animals, labelling strongly the target cells and particularly the nuclear Cowdry type A inclusion body, which is the most diagnostic characteristics of this disease.

Animals↗

PubMed2005

Spiroplasma penaei sp. nov., associated with mortalities in Penaeus vannamei, Pacific white shrimp.

A new bacterial strain, designated SHRIMP(T), isolated from the haemolymph of the Pacific white shrimp, Penaeus vannamei, was serologically distinct from other spiroplasmas. Cells of this strain were helical in form and variable in length. Examination by electron microscopy revealed wall-less cells delineated by a single cytoplasmic membrane. The organisms grew well in M1D media supplemented with 2 % NaCl. Strain SHRIMP(T) grew at temperatures of 20-37 degrees C, with optimum growth occurring at 28 degrees C. The strain catabolized glucose and hydrolysed arginine, but did not hydrolyse urea. The G+C content of the DNA was 29 +/-1 mol%. Strain SHRIMP(T) (=ATCC BAA-1082T=CAIM 1252T) is designated the type strain of a novel species, Spiroplasma penaei sp. nov., which represents a new subgroup (I-9) of the group I spiroplasmas.

Animals↗

PubMed2006

The complete analysis of oxytetracycline pharmacokinetics in farmed Pacific white shrimp (Litopenaeus vannamei).

Lack of dosing information of the major antibiotics known as oxytetracycline (OTC) for the Pacific white shrimp (Litopenaeus vannamei) could have harmful impact on aquaculture in Thailand. The aim of this study was to detail complete pharmacokinetic information of OTC in the Pacific white shrimp. Sixty-four male L. vannamei weighing 14-22 g with carapace length of 2.30-3.00 cm in the standardized moulting stage of C-D(0) were used for the investigations. Single dose, 10 microg/g body weight OTC solution was administered intra-sinusally (i.s.), and the shrimps were then sampled in three replicates at time intervals of 0.25, 0.5, 2, 4, 6, 9, 12, 24, 48, 72, 170, 336 and 504 h postdose. OTC levels with time intervals in biological matrices including the hemolymph, abdominal muscle, and digestive gland of each sample were determined by validated high-performance liquid chromatography, and were analyzed with noncompartment and compartment models. A simplified two-compartment model was employed rather than a more complicated model, with additional digestive compartment if necessary. A significant portion of the OTC was found in the digestive glands, even though the OTC was administered i.s. The model indicated that the OTC was thus not only distributed into the tissue compartment, but also to the digestive gland, from where it was eliminated from the shrimp's body. The dispositional half-lives of all compartments was found to be 14-21 h. Approximately 60% of the drug elimination took place in digestive gland, which is proposed to be the major route of elimination.

Animals↗

PubMed2006

Cryptic biodiversity and phylogeographical patterns in a snapping shrimp species complex.

Recent investigations suggest that marine biodiversity may be much higher than earlier estimates, and an important hidden source of diversity in marine systems is the phenomenon of cryptic species complexes. Such complexes are informative models for research into the evolutionary processes that govern species compositions of marine fauna. The snapping shrimp genera Alpheus and Synalpheus are known to harbour large numbers of cryptic species; here, I characterize the genetic structure of the Alpheus armillatus species complex in the northern Caribbean, west Atlantic, and Gulf of Mexico using mitochondrial and nuclear sequence data. Over this geographical region, the complex harbours at least three lineages that are probable reproductively isolated species; all major lineages diverged subsequent to the close of the Isthmus of Panama. Only one lineage was present in the Gulf of Mexico, whereas outside the Gulf of Mexico there was no clear tendency for lineage dominance by geographical region, as most sites were populated by shrimp from at least two lineages. However, within each lineage, there was strong evidence of population genetic differentiation between geographical regions. All lineages showed strong signals of demographic expansion, and one lineage showed sharply reduced genetic diversity, suggestive of past population bottlenecks or recently founded populations with low gene flow from other sites. These results show that evolutionary processes leading to divergence and speciation have been common and recent in the snapping shrimp, and suggest that connectivity among shrimp populations may be limited.

Americas↗

PubMed2006

Phenotypic and molecular typing of Vibrio harveyi isolates and their pathogenicity to tiger shrimp larvae.

AIMS: The objective of the present study was to identify the biotype(s) and molecular type(s) of Vibrio harveyi associated with pathogenicity in tiger shrimp (Penaeus monodon) larvae. METHODS AND RESULTS: Five luminescent and four nonluminescent V. harveyi isolates were subjected to phenotyping and random amplified polymorphic DNA (RAPD) fingerprinting, and pathogenicity testing to P. monodon mysis. Four isolates induced 34-41% mortality of P. monodon mysis when challenged at the rate of 10(6) CFU ml(-1) within 60 h. Sucrose-fermenting biotypes of V. harveyi appeared to be associated with pathogenicity to larval shrimp. Higher temperature and salinity appeared to play a role on the onset of vibriosis and mortality in the challenged larval shrimp. Pathogenic isolates of V. harveyi could be demarcated as revealed by their clustering in the dendrogram constructed based on the RAPD fingerprints. CONCLUSIONS: Nonluminescent V. harveyi also appear to be important aetiological agents of vibriosis of shrimp larvae. Sucrose-fermenting biotypes are likely to be pathogenic. High temperature may trigger onset of vibriosis. SIGNIFICANCE AND IMPACT OF THE STUDY: Biotyping of V. harveyi isolates and looking for traits, such as ability to ferment sucrose may be helpful in identifying the pathogenic forms, and such approach requires to be investigated further with larger number of isolates.

Animals↗

PubMed1980

Microbiological quality of frozen shrimp and lobster tail in the retail market.

The microbiological quality of three frozen shrimp products and frozen lobster tail at the retail level was determined. The number of retail units of the four products examined and the geometric means for aerobic plate counts at 30 and 35 degrees C, respectively, were: 1,464 units of cooked, peeled shrimp--13,000 and 7,200 per g; 1,468 units of raw, peeled shrimp--860,000 and 300,000 per g; 1,300 units of raw, in-shell shrimp--800,000 and 300,000 per g; 1,315 units of lobster tail--140,000 and 42,000 per g. Geometric means for coliform, Escherichia coli, and Staphylococcus aureus counts for all products were < 10 per g.

Decapoda↗

PubMed1998

Cloning and nucleotide sequence of the gyrB gene of Vibrio parahaemolyticus and its application in detection of this pathogen in shrimp.

Because biochemical testing and 16S rRNA sequence analysis have proven inadequate for the differentiation of Vibrio parahaemolyticus from closely related species, we employed the gyrase B gene (gyrB) as a molecular diagnostic probe. The gyrB genes of V. parahaemolyticus and closely related Vibrio alginolyticus were cloned and sequenced. Oligonucleotide PCR primers were designed for the amplification of a 285-bp fragment from within gyrB specific for V. parahaemolyticus. These primers recognized 117 of 117 reference and wild-type V. parahaemolyticus strains, whereas amplification did not occur when 90 strains of 37 other Vibrio species or 60 strains representing 34 different nonvibrio species were tested. In 100-microliter PCR mixtures, the lower detection limits were 5 CFU for live cells and 4 pg for purified DNA. The possible application of gyrB primers for the routine identification of V. parahaemolyticus in food was examined. We developed and tested a procedure for the specific detection of the target organism in shrimp consisting of an 18-h preenrichment followed by PCR amplification of the 285-bp V. parahaemolyticus-specific fragment. This method enabled us to detect an initial inoculum of 1.5 CFU of V. parahaemolyticus cells per g of shrimp homogenate. By this approach, we were able to detect V. parahaemolyticus in all of 27 shrimp samples artificially inoculated with this bacterium. We present here a rapid, reliable, and sensitive protocol for the detection of V. parahaemolyticus in shrimp.

Amino Acid Sequence↗

PubMed2005

RING-H2 protein WSSV249 from white spot syndrome virus sequesters a shrimp ubiquitin-conjugating enzyme, PvUbc, for viral pathogenesis.

Modification of proteins by ubiquitin is essential for numerous cellular processes. The RING-H2 finger motif has been implicated in ubiquitin-conjugating enzyme (E2)-dependent ubiquitination. Four proteins, WSSV199, WSSV222, WSSV249, and WSSV403, from white spot syndrome virus (WSSV) contain the RING-H2 motif. Here we report that WSSV249 physically interacts with a shrimp ubiquitin-conjugating enzyme, PvUbc, and mediates ubiquitination through its RING-H2 motif in the presence of E1 and PvUbc. Mutations of the putative zinc coordination residues in the RING-H2 domain of WSSV249, however, ablate ubiquitination efficiency. In addition, the RING-H2 domain of WSSV249 is capable of ubiquitination with UbcH1, UbcH2, UbcH5a, UbcH5b, UbcH5c, UbcH6, and UbcH10, respectively, exhibiting a low degree of E2 specificity. Significantly, the expression of WSSV249 and PvUbc increased during infection, as revealed by real-time PCR. Furthermore, in situ hybridization showed that WSSV249 and PvUbc display similar expression patterns in infected shrimps, and immunofluorescence and immunohistochemistry assays showed an increase of PvUbc in infected shrimp cells. These results suggest that the RING-H2 protein WSSV249 from WSSV may function as an E3 ligase via sequestration of PvUbc for viral pathogenesis in shrimp.

Amino Acid Sequence↗

PubMed1987

Soy- and shrimp-specific IgE responses in orally and intraperitoneally immunized mice.

Female C3H/HeJ mice were immunized parenterally and/or intragastrically with two important food allergens, soy and shrimp. Although both preparations elicited specific IgE production when administered intraperitoneally, anti-shrimp titers were consistently higher. Soy or shrimp administration, i.g. with Pertussis adjuvant (intraperitoneally or intragastrically) did not induce detectable reaginic antibody. Animals treated with soy intragastrically were unable to produce a soy-specific IgE response upon intraperitoneal immunization. The combination of soy and Pertussis (intragastric) did, however, abrogate this tolerizing effect on IgE synthesis. In contrast to soy, the shrimp antigens (intragastric) did not act as tolerogens and were found to enhance subsequent reaginic responses produced by intraperitoneal immunization. These results are considered with respect to mechanisms operative in food allergy.

Adjuvants, Immunologic↗

PubMed1992

Microscopic and ultrastructural studies of necrotizing hepatopancreatitis in Pacific white shrimp (Penaeus vannamei) cultured in Texas.

Granulomatous hepatopancreatitis of unknown etiology has been considered an important disease of Texas shrimp mariculture since 1985. Samples of Pacific white shrimp (Penaeus vannamei) were collected during 1986, 1987, and 1990 from three farms and an experimental mariculture facility with histories of production loss and increased mortality rates. Histologic and ultrastructural examination of shrimp from the four sites demonstrated two morphologically distinct, Gram-negative, double-enveloped, intracytoplasmic bacteria in necrotic hepatopancreatic epithelium. The more numerous small, pleomorphic rod as well as the helical organism are both taxonomically unclassifiable. The helical organism lacked ultrastructural characteristics of previously described helical or spiral bacteria. The relationship between the two organisms is unknown, but the pleomorphic rod is thought to play a major role in the disease. The role of a bacterial agent(s) in subsequent disease episodes is suggested by the observation that the use of oxytetracycline-medicated feed resulted in increased production and survival. Hepatopancreatic tubular epithelial necrosis and shrimp mortality correlated directly with the extent of infection by the small pleomorphic rod. Individual discrete bacteria were identified microscopically by Steiner and Steiner's method. Three major developmental stages of the disease were characterized based on the extent and number of hepatopancreatic tubular epithelial cells containing bacteria, the degree of tubular interstitial inflammation, and the extent and chronicity of tubular necrosis. Additional studies are needed to clarify the roles of the different bacteria identified and the potential role of environmental factors on the disease process.

Animals↗

PubMed2025

Population stratification in Northern shrimp (Pandalus borealis) off Iceland evident from RADseq analysis.

The northern shrimp Pandalus borealis (ice. Stóri kampalampi) is a North Atlantic crustacean of significant commercial interest which has been harvested consistently in Icelandic waters since 1936. In Icelandic waters, the length at which this protandrous species transitions from male to female differs between the inshore and offshore populations, suggesting a biologically meaningful stratification which may or may not be plastic. Using reduced representative genomes assembled from RADseq data, sampled from 96 individuals collected at two time points (2018 and 2021), we compare the level of genetic structure across a gradient extending out of Skjálfandi bay, north Iceland. These data are compared to samples from a far offshore site, some 65 km out from the bay, as well as another inshore fjord in Arnarfjörður, in northwestern Iceland. Since 1999, no harvesting of inshore populations of P. borealis in Skjálfandi has been allowed due to stock decline, but harvesting of offshore stocks has continued. Uncertainty surrounding the extent of structure between the in- and offshore aggregations has remained. Here we report distinct genetic structure defining the inshore and offshore populations of northern shrimp, but find significant admixture between the two. Most importantly, we see that genetically inshore populations of northern shrimp extend far outside the harvest boundaries of inshore shrimp, and offshore individuals may exhibit punctuated migration into the inshore areas.

Animals↗

PubMed2005

Extreme impact and cavitation forces of a biological hammer: strike forces of the peacock mantis shrimp Odontodactylus scyllarus.

Mantis shrimp are renowned for their unusual method of breaking shells with brief, powerful strikes of their raptorial appendages. Due to the extreme speeds of these strikes underwater, cavitation occurs between their appendages and hard-shelled prey. Here we examine the magnitude and relative contribution of the impact and cavitation forces generated by the peacock mantis shrimp Odontodactylus scyllarus. We present the surprising finding that each strike generates two brief, high-amplitude force peaks, typically 390-480 micros apart. Based on high-speed imaging, force measurements and acoustic analyses, it is evident that the first force peak is caused by the limb's impact and the second force peak is due to the collapse of cavitation bubbles. Peak limb impact forces range from 400 to 1501 N and peak cavitation forces reach 504 N. Despite their small size, O. scyllarus can generate impact forces thousands of times their body weight. Furthermore, on average, cavitation peak forces are 50% of the limb's impact force, although cavitation forces may exceed the limb impact forces by up to 280%. The rapid succession of high peak forces used by mantis shrimp suggests that mantis shrimp use a potent combination of cavitation forces and extraordinarily high impact forces to fracture shells. The stomatopod's hammer is fundamentally different from typical shell-crushing mechanisms such as fish jaws and lobster claws, and may have played an important and as yet unexamined role in the evolution of shell form.

Acoustics↗