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At least 343 records · Page 19Linked to original sources
PubMed2000

Enzymatic and osmoregulative alterations in white shrimp Litopenaeus vannamei exposed to pesticides.

Pesticide pollution in coastal ecosystems of Sinaloa, Mexico is considered to be a cause for slow growth, increase of diseases and sometimes massive mortality of shrimp. So it was necessary to develop fast techniques to detect pesticide pollution in shrimp habitats. Enzymatic and osmoregulation tests in shrimp exposed to DDT, Lindane, Chlordane, Lorsban, Gusathion, Folidol, Diazinon and Tamaron were carried out. Activity reductions from 11 to 2 units/ml in acetylcholinesterase and from 1 to 0 units/l in transaminases (GOT and GPT) were detected. Also increases in osmoregulation were observed in shrimp exposed to Folidol, Diazinon and Gusation, whereas decreases with DDT, Lindane and Lorsban at salinity 50/1000. We conclude that pesticides are causing alterations in these biochemical functions and this kind of tests represent a rapid and inexpensive method for pesticide pollution detection.

Acetylcholinesterase↗
PubMed2000

Introducing foreign DNA into tiger shrimp (Penaeus monodon) by electroporation.

Electroporation was used to introduce pFLAG-CMV-1-BAP, a DNA fragment that includes a bacterial alkaline phosphatase gene driven by a human cytomegalovirus (CMV) promoter, into Penaeus monodon zygotes. The transgenic tiger shrimp was achievedby using 10kV, 28 pulses, 120 g sec pulse time, 10 cycles, and a DNA concentration of 37.5 microg/mL. The hatching rate of electroporated zygotes (46%) was significantly lower than that of zygotes in the untreated group (89%). The survival rate of postlarvae in the electroporated group using a DNA concentration of 37.5 microg/mL decreased from 0.6% for postlarva 45 to 0.4% for postlarva 120. Based on dot blot analysis, the rate of gene transfer was 37% in mysis-stage, 23% postlarva 15(PL15), 19% postlarva 45(PL45), and 21% 4-month-old (about PL120). Genomic Southern blotting demonstrated that DNA from transgenic tiger shrimp contained fragments of exogenous DNA that were smaller, larger and of the same molecular size as pFLAG-CMV-1-BAP. Transferred DNA fragments were integrated into the genomes of 31% of the transgenic tiger shrimp. The exogenous DNA was mosaically distributed in a wide variety of tissues. Immunohistochemical staining revealed that the FLAG-BAP fused-protein encoded by pFLAG-CMV-1-BAP was present in the ovaries of some transgenic tiger shrimp.

Alkaline Phosphatase↗
PubMed2001

Effect of initial glucose concentration and inoculation level of lactic acid bacteria in shrimp waste ensilation.

Fermentation conditions and microorganisms were determined, based on acid production, glucose concentration as carbohydrate source. Inoculation levels to obtain a stable shrimp waste silage were also determined. Shrimp waste ensilation was an efficient method of preservation, allowing the recovery of chitin and another added-value products such as pigments, proteins and enzymes. From the various lactic acid bacteria tested, Lactobacillus pentosus and Lactobacillus sp. (B2) were the best lactic acid producers, although small quantities of acetic acid were detected in samples inoculated with Lactobacillus pentosus. Therefore B2 was chosen for the analysis of glucose consumption as well as for the determination of optimum inoculation levels. The best results were obtained at 10% (w/w wet basis) and 5% (v/w wet basis) respectively. Presence of starters and initial glucose concentration were critical factors in the fermentation of shrimp waste. High initial glucose and starter concentrations reduced the time and increased the amount of lactic acid produced. The fermentation pattern changed during ensilation from hetero to homofermentative. Shrimp waste ensilation prevented the growth of spoilage microorganisms keeping their microbial counts steady and pH values within the acid region.

Journal Article↗
PubMed1999

Molecular cloning and characterization of prophenoloxidase in the black tiger shrimp, Penaeus monodon.

A cDNA encoding shrimp, Penaeus monodon, prophenoloxidase (proPO) was obtained by screening a hemocyte library by plaque hybridization using a proPO cDNA fragment from freshwater crayfish, Pacifastaceus leniusculus, as a probe. The 3,002 bp cDNA contains an open reading frame of 2,121 bp and a 881 bp 3'-untranslated region. The molecular mass of the deduced amino acid sequence (688 amino acids) is 78,700 Da with an estimated pI of 5.8. Two putative copper binding sites are present and they have a highly conserved sequence around these sites. No signal peptide was detected in the shrimp proPO, as has been previously shown to be the case for all arthropod proPOs cloned so far. The cleavage site of zymogen activation is likely to be between Arg 44 and Val 45. A tentative complement-like motif (GCGWPQHM) is also present. Shrimp proPO mRNA is synthesized in the hemocytes and not in the hepatopancreas. Comparison of amino acid sequences showed that shrimp proPO is more closely related to another crustacean proPO, namely crayfish, than to the insect proPOs.

Amino Acid Sequence↗
PubMed2003

Chronic toxicity of arsenic to the Great Salt Lake brine shrimp, Artemia franciscana.

We determined the chronic toxicity of arsenic (sodium arsenate) to the Great Salt Lake brine shrimp, Artemia franciscana. Chronic toxicity was determined by measuring the adverse effects of arsenic on brine shrimp growth, survival, and reproduction under intermittent flow-through conditions. The study commenced with <24-h-old nauplii, continued through reproduction of the parental generation, and ended after 28 days of exposure. The concentrations tested were 4, 8, 15, 31, and 56mg/L dissolved arsenic. The test was conducted using water from the Great Salt Lake, Utah as the dilution water. Adult survival was the most sensitive biological endpoint, with growth and reproduction somewhat less sensitive than survival. The no observed effect concentration (NOEC) for survival was 8mg/L, and the lowest observed effect concentration (LOEC) was 15mg/L dissolved arsenic. The LOEC for growth and reproduction was greater than the highest concentration tested, 56mg/L. Based on survival, the final chronic value (geometric mean of the NOEC and LOEC) was 11mg/L dissolved arsenic. The F(1) generation appeared to acclimate to the prior arsenic exposure of the parental generation and was significantly less sensitive than the parental generation. For example, survival for the F(1) generation through day 12 was 100% in 56mg/L dissolved arsenic, compared to 26% for the parental generation. Growth of the F(1) generation was significantly less than that of the parental generation across all concentrations including the control, indicating a generational difference in brine shrimp growth rather than an arsenic effect. This study represents one of the few full life cycle toxicity tests conducted with brine shrimp.

Animals↗
PubMed2002

Quantitative assay for measuring the Taura syndrome virus and yellow head virus load in shrimp by real-time RT-PCR using SYBR Green chemistry.

Taura syndrome virus (TSV) and yellow head virus (YHV) are the two RNA viruses infecting penaeid shrimp (Penaeus sp.) that have caused major economic losses to shrimp aquaculture. A rapid and highly sensitive detection and quantification method for TSV and YHV was developed using the GeneAmp 5700 Sequence Detection System and SYBR Green chemistry. The reverse transcriptase polymerase chain reaction (RT-PCR) mixture contained a fluorescent dye, SYBR Green, which exhibits fluorescence enhancement upon binding to double strand cDNA. The enhancement of fluorescence was found to be proportional to the initial concentration of the template cDNA. A linear relationship was observed between input plasmid DNA and cycle threshold (C(T)) values for 10(6) down to a single copy of both viruses. To control for the variation in sample processing and in reverse transcription reaction among samples, shrimp beta-actin and elongation factor-1alpha (EF-1alpha) genes were amplified in parallel with the viral cDNA. The sensitivity and the efficiency of amplification of EF-1alpha was greater than beta-actin when compared to TSV and YHV amplification efficiency suggesting that EF-1alpha is a better internal control for the RT-PCR detection of TSV and YHV. In addition, sample to sample variation in EF-1alpha C(T) value was lower than the variation in beta-actin C(T) value of the corresponding samples. The specificity of TSV, YHV, EF-1alpha and beta-actin amplifications was confirmed by analyzing the dissociation curves of the target amplicon. The C(T) values of TSV and YHV samples were normalized against EF-1alpha C(T) values for determining the absolute copy number from the standard curve of the corresponding virus. The method described here is highly robust and is amenable to high throughput assays making it a useful tool for diagnostic, epidemiological and genetic studies in shrimp aquaculture.

Actins↗
PubMed1997

Detection of yellowhead virus and Chinese baculovirus in penaeid shrimp by the Western blot technique.

The continuing threat posed by viral diseases in cultured shrimp calls for the development of detection technologies for monitoring the animals, especially broodstock. Two of the most highly pathogenic viruses of penaeid shrimp are the yellow-head virus (YHV) and Chinese baculovirus (CBV, also called white spot baculovirus). A Western blot (WB) protocol capable of detecting YHV and CBV in the hemolymph of infected shrimp was developed. The use of the hemolymph as material for virus detection allowed for sample collection without sacrificing the animals. This protocol was highly specific, rapid, and sensitive enough to detect the presence of the viruses before the appearance of overt symptoms. It was also useful for demonstrating the growth of both viruses in primary shrimp lymphoid cell cultures.

Animals↗
PubMed2002

Reproductive alterations in adult grass shrimp, Palaemonetes pugio, following sublethal, chronic endosulfan exposure.

Grass shrimp (Palaemonetes pugio) populations exposed to anthropogenic contaminant sources in South Carolina (SC) have reduced densities when compared with populations at SC-reference sites. This laboratory study examined the effects of a commonly used agricultural insecticide, endosulfan, on grass shrimp reproduction. Reproductively active grass shrimp were chronically exposed to sublethal concentrations of endosulfan (200 or 400 ng/l) for 43 days. The cumulative number of females that became gravid and the rate at which they became gravid were measured. Endosulfan exposure reduced the cumulative number of gravid females by 31% in the 200 ng/l exposure and 39% in the 400 ng/l exposure. The first appearance of gravid females in the population was significantly delayed in treated populations compared with the control treatment in a dose dependent manner. Clutch size in these gravid females was not significantly different among the treatments. Additionally, there was no difference in the onset of reproduction in the treated populations. These results implicate a population reduction due to a decrease in the overall number of females becoming gravid in a population over time, not a reduction in clutch size per individual. While the mechanisms of action have yet to be defined, these results indicate that sublethal endosulfan concentrations may have a negative effect on grass shrimp reproductive biology.

Animals↗
PubMed2003

The potential use of constructed wetlands in a recirculating aquaculture system for shrimp culture.

A pilot-scale constructed wetland unit, consisting of free water surface (FWS) and subsurface flow (SF) constructed wetlands arranged in series, was integrated into an outdoor recirculating aquaculture system (RAS) for culturing Pacific white shrimp (Litopenaeus vannamei). This study evaluated the performance of the wetland unit in treating the recirculating wastewater and examined the effect of improvement in water quality of the culture tank on the growth and survival of shrimp postlarvae. During an 80-day culture period, the wetland unit operated at a mean hydraulic loading rate of 0.3 m/day and effectively reduced the influent concentrations of 5-day biochemical oxygen demand (BOD5, 24%), suspended solids (SS, 71%), chlorophyll a (chl-a, 88%), total ammonium (TAN, 57%), nitrite nitrogen (NO2-N, 90%) and nitrate nitrogen (NO3-N, 68%). Phosphate (PO4-P) reduction was the least efficient (5.4%). The concentrations of SS, Chl-a, turbidity and NO3-N in the culture tank water in RAS were significantly (P<or=.05) lower than those in a control aquaculture system (CAS) that simulated static pond culture without wetland treatment. However, no significant difference (P<or=.05) in BOD5, TAN and NO2-N was found between the two systems. At the end of the study, the harvest results showed that shrimp weight and survival rate in the RAS (3.8 +/-1.8 g/shrimp and 90%) significantly (P<or=.01) exceeded those in the CAS (2.3+/-1.5 g/shrimp and 71%). This study concludes that constructed wetlands can improve the water quality and provide a good culture environment, consequently increasing the shrimp growth and survival without water exchange, in a recirculating system.

Animals↗
PubMed1998

Shrimp plasma HDL and beta-glucan binding protein (BGBP): comparison of biochemical characteristics.

A high density lipoprotein (HDL) and beta-glucan binding protein (BGBP) have been found in the hemolymph of marine shrimp. These proteins are involved in the transport of lipid and the recognition of foreign matter, respectively. Similarities in the color of the proteins and the molecular mass were noted. For a detailed comparison, HDL and BGBP were purified from two shrimp species, Penaeus vannamei and Penaeus californiensis, and their biochemical characteristics determined. Both proteins from each of the shrimp species are monomeric with approximately the same molecular mass in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (approximately 100-112 kDa) and contain carbohydrate and lipid. The amino acid composition is similar and there is a high degree of similarity in the N-terminus. Furthermore, they are recognized by antibodies prepared independently. These results reveal that BGBP and HDL in shrimp hemolymph are the same protein, suggesting that there is a close relationship between the ability to respond to foreign matter and the diet as a provider of essential nutrients.

Amino Acid Sequence↗
PubMed1998

Confirmation of multiple tetracycline residues in milk and oxytetracycline in shrimp by liquid chromatography-particle beam mass spectrometry.

A confirmation procedure is described for detection of residues of six tetracyclines in bovine milk, and oxytetracycline in shrimp. Residues are extracted from milk or shrimp tissue using metal chelate affinity chromatography. The extracts are desalted, further concentrated using polymeric solid-phase extraction, and chromatographed on a polymeric reversed-phase column. Analysis is by methane negative ion chemical ionization on a quadrupole mass spectrometer using a particle beam interface. Data are acquired in partial scan mode, monitoring from m/z 378 to m/z 480. The procedure was validated with control milk and shrimp, fortified milk (30 ng/ml) and shrimp (100 ng/g), and milk and tissue from animals treated with the drugs.

Animals↗
PubMed2000

Purification of vitellin from grass shrimp Palaemonetes pugio, generation of monoclonal antibodies, and validation for the detection of lipovitellin in Crustacea.

Much effort has been put into developing vitellogenin antibodies against a wide variety of aquatic vertebrate species to study potential estrogen or anti-estrogen endocrine disrupters. Little work has been done on endocrine disruption in aquatic invertebrates. Although some antibodies have been produced against blue crab and penaeid shrimp lipovitellin, they have only poor cross-reactivity with the important estuarine grass shrimp, Palaemonetes pugio. Vitellin was purified from eggs, monoclonal antibodies were produced using standard techniques, and hybridoma supernatants were screened by ELISA. Western blots were done using extracts from male and female grass shrimp to verify specificity of the monoclonal antibodies. Two low molecular mass bands in the range of 68-85 kD and two high molecular mass bands in the range of 190-221 kD were found. In addition to grass shrimp, several other crustacean species were screened and cross-reactivity found, including blue crab (Callinectes sapidus), mud crab (Rhithropanopeus harrisii), red swamp crayfish (Procambarus clarkii ) and Daphnia magna. To further investigate the use of the antibody, we performed a chronic 6-week pyrene exposure study. We found that vitellin was upregulated in females after 6 weeks and that this may be a protective measure against lipophilic xenobiotics.

Animals↗
PubMed2000

Is the brown shrimp Crangon crangon (L.) population of the Vaccarès lagoon (Camargue, France, Rhône delta) an annual population?

To study the population dynamics of Crangon crangon in the Vaccarès lagoon in the Camargue (Rhône delta), the age of brown shrimp was determined using the method of Tiews (1954). This method consists in counting the number of segments of the outer branch of the first antenna. This number is correlated with the number of moults already accomplished by the shrimp. The duration of the intermoult depends on water temperature. Adapting this model to our environmental data, we determined the age of the brown shrimp. Hatching of larvae occurs in the sea in May, and juveniles enter the lagoon from June to September. Our results showed that Crangon crangon in the Vaccarès lagoon reached sexual maturity between 6 and 11 months, and individuals are 1 year old when they leave the lagoon. A few adults from the previous year (1+) migrate back to the lagoon the following year, but these older individuals represent only a very small part of the population, and have only been observed occasionally. The Vaccarès brown shrimp population can therefore be considered as annual, in contrast to northern European populations.

Animals↗
PubMed2001

Digestive proteinases of red shrimp Pleoticus muelleri (Decapoda, Penaeoidea): partial characterization and relationship with molting.

The present study describes the activity and some characteristics of proteinases in the hepatopancreas of red shrimp Pleoticus muelleri during the different stages of the molting cycle. Proteolytic activity was highest between pH 7.5 and 8. The hepatopancreatic protein content in the premolt stage was higher than in the other stages of the molting cycle (P<0.05). No significant differences were found in total proteolytic activity in the hepatopancreas when comparing molting stages. The proteolytic activity of the P. muelleri hepatopancreas enzyme preparations is the main responsibility of serine proteinases. TLCK, a trypsin inhibitor, reduced azocasein hydrolysis between 26% (intermolt) and 37% (premolt). TPCK, a chymotrypsin inhibitor, did not decrease hydrolytic activity, except for in postmolt. Low trypsin and chymotrypsin activities were found during intermolt, and increased in postmolt. The electrophoretogram of the enzyme extracts shows 12 bands of activity during intermolt (from 16.6 to 53.1 kDa). Some fractions were not detected in the postmolt and premolt stages. Three low molecular weight trypsin forms (17.4, 19.1 and 20 kDa) were found in all molting stages. One band of chymotrypsin (21.9 kDa) was observed in all molting stages. High molecular mass active bands (66-205 kDa) could not be characterized with inhibitors. Comparison of the protease-specific activity of the hepatopancreas of some species indicated a relationship between digestive enzyme activity and feeding habits of the shrimp. Omnivorous shrimp, such as Penaeus vannamei (syn: Litopenaeus vannamei) and Penaeus monodon, showed higher protease activity than the carnivorous shrimp, Penaeus californiensis (syn: Farfantepenaeus californiensis) and P. muelleri. In fact, the enzymatic activity in the hepatopancreas of P. muelleri showed variations in relation to feeding habit and molting cycle.

Animals↗
PubMed2003

Effects of dietary protein on the activity and mRNA level of trypsin in the midgut gland of the white shrimp Penaeus vannamei.

Protein food modulates the activity of proteases of the midgut gland of Penaeus vannamei. Shrimp fed with food containing 15, 30 and 50% protein exhibited differences in trypsin and chymotrypsin activity and trypsin mRNA levels. Shrimp fed with 30% protein showed higher trypsin and chymotrypsin activities than those fed 15 or 50% protein. An additional paralogue trypsin was observed with electrophoretic analysis in shrimp fed 30% protein. Shrimp fed 30% protein showed the highest trypsin to mRNA concentration, suggesting that trypsin genes expression is regulated transcriptionally.

Animals↗
PubMed2003

Purification and characterization of alpha 2-macroglobulin from the white shrimp (Penaeus vannamei).

alpha(2)-Macroglobulin (alpha(2)M) is a broad-spectrum protease-binding protein abundant in plasma from vertebrates and several invertebrate phyla. This protein was purified from cell-free hemolymph of the white shrimp, Penaeus vannamei, using Blue-Sepharose and Phenyl-Sepharose chromatography. The shrimp alpha(2)M is a 380 kDa protein, a homodimer of two apparently identical subunits of approximately 180 kDa linked by disulphide bridges. The amino acid sequence of the N-terminus is similar to the Limulus alpha(2)M counterpart. The shrimp alpha(2)M has a wide inhibition spectrum against different proteinase types including trypsin, leucine amino peptidase, chymotrypsin, elastase and papain. The secondary structure of shrimp alpha(2)M is mainly beta-sheet (36%), with a characteristic minimum elipticity at 217 nm. Evidence for a thiolester-mediated inhibition mechanism of proteases by alpha(2)M was provided by inactivation with methylamine.

Amino Acid Sequence↗
PubMed2003

Identification of lactic acid bacteria from spoilage associations of cooked and brined shrimps stored under modified atmosphere between 0 degrees C and 25 degrees C.

AIMS: To evaluate spoilage and identify lactic acid bacteria (LAB) from spoilage associations of cooked and brined shrimps stored under modified atmosphere packaging (MAP) at 0, 5, 8, 15 and 25 degrees C. METHODS AND RESULTS: Bacterial isolates (102) from spoilage associations of cooked and brined MAP shrimps were characterized by phenotypic tests and identified as lactic acid bacteria (78 isolates), other Gram-positive bacteria (13 isolates) and Gram-negative bacteria (11 isolates). A selection of 48 LAB isolates were further characterized and identified by phenotypic tests and SDS-PAGE electrophoresis of whole cell proteins. Selected clusters of LAB isolates were analysed by plasmid profiling, pulsed field gel electrophoresis and 16S rRNA gene sequencing. Enterococcus faecalis was identified in spoilage associations at 15 degrees C and 25 degrees C, and its metabolic activity corresponded to chemical changes in spoiled products. Carnobacterium divergens, a non-motile Carnobacterium sp. nov. and Lactobacillus curvatus were the LAB species observed in spoilage associations of products stored at 0 degrees C, 5 degrees C and 8 degrees C. CONCLUSIONS: Enterococcus spp. and Carnobacterium spp. were the dominant parts of spoilage associations of cooked and brined MAP shrimps stored at high and low temperatures, respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: The SDS-PAGE technique and simple biochemical keys allowed the majority of LAB isolates from spoilage associations of cooked and brined MAP shrimps to be identified at the species level.

Animals↗
PubMed2003

Population genetic analysis of white shrimp, Litopenaeus setiferus, using microsatellite genetic markers.

The white shrimp (Litopenaeus setiferus) is a commercially and recreationally valuable species, yet little is known of its population structure or genetic diversity. White shrimp are distributed along the Atlantic coast of the United States and from the west coast of Florida to the Bay of Campeche, Mexico. In this study, shrimp were collected from North Carolina, South Carolina (four separate collections were taken from 1995 to 1999), Georgia, the Atlantic and Gulf coasts of Florida, Louisiana, Texas and Mexico. DNA was isolated from these individuals, and genetic variation was assessed at six microsatellite loci. These loci were, for the most part, highly polymorphic with an average expected heterozygosity of 0.68. Deviations from Hardy-Weinberg proportions were observed over all samples, but experimental results suggested the presence of null alleles, which confounded a biological interpretation of this result. Pairwise tests of the similarity of allele frequency distributions and distance measure analyses showed broad-scale genetic homogeneity superimposed over occasional indications of random geographical and temporal differentiation. FST and RST estimates over all loci and samples were 0.002 or less and indicated little population structure. Weak but significant genetic differentiation was evident only between pooled western Atlantic and pooled Gulf of Mexico samples. Within the Gulf of Mexico or within the western Atlantic, the large-scale genetic homogeneity observed may be a consequence of genetic mixing resulting from pelagic larvae and adult migrations, while the random local genetic differentiation may be a result of genetic sampling or experimental sampling error. The weak differentiation between shrimp from the Gulf of Mexico and the western Atlantic can be explained by a relatively recent separation of these two populations and/or small amounts of ongoing gene flow.

Animals↗