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At least 325 records · Page 18Linked to original sources

PubMed2006

White shrimp (Litopenaeus vannamei) recombinant lysozyme has antibacterial activity against Gram negative bacteria: Vibrio alginolyticus, Vibrio parahemolyticus and Vibrio cholerae.

C-type lysozyme has been described as an antibacterial component of the shrimp innate defence system. We determined quantitatively the antibacterial activity of white shrimp (Litopenaeus vannamei) recombinant lysozyme against three Gram negative bacteria: Vibrio alginolyticus, Vibrio parahemolyticus and Vibrio cholerae, using a turbidimetric assay with live bacteria and differential bacterial viable count after interaction with the protein. In conclusion, the antibacterial activity of recombinant shrimp lysozyme against Vibrio sp. is at least equal to the values against the Gram positive M. luteus and more active against the shrimp pathogens V. alginolyticus and V. parahemolyticus.

Animals↗

PubMed2006

Molecular cloning and tissue distribution of ferritin in Pacific white shrimp (Litopenaeus vannamei).

Ferritin, a cytosolic iron storage protein composed of 24 subunits of heavy chain and light chain, is an intracellular protein primarily involved in iron metabolism. It can sequester up to 4500 ferric ions in its inner core to protect cells against toxicity of iron. Ferritin is known to play important roles in detoxification and is also involved in immunity processes. In this study, a full-length ferritin cDNA was cloned from the haemocyte of the Pacific white shrimp, Litopenaeus vannamei: it comprises 1249 bp, including 132 bp in the 5'-untranslated region, 510 bp in the open reading frame which encodes 170 amino acid residues, and 607 bp in the 3'-untranslated region. Alignments of the deduced amino acid sequence showed that the Pacific white shrimp ferritin shares 74%, 69%, 62%, 67%, 50% and 48% identity with crayfish, tick, brine shrimp, oyster, human and rat, respectively. The tissue-specific expression pattern was examined by reverse transcription polymerase chain reaction and real-time quantitative PCR. The ferritin mRNA is expressed in various tissues of the shrimp in the order of haemocyte, midgut gland, brain ganglion, gill, hepatopancreas, abdominal ganglion, eyestalk, muscle, thoracic ganglion, and heart.

Amino Acid Sequence↗

PubMed2006

Cloning and expression analysis of an o-methyltransferase (OMT) gene from Chinese shrimp, Fenneropenaeus chinensis.

O-methyltransferase (OMT) is ubiquitously present in diverse organisms and plays an important regulatory role in plant and animal growth, development, reproduction and defence and has also been implicated in human emotion and disease. A putative o-methyltransferase (OMT) gene has been cloned from the haemocytes of bacteria-infected Chinese shrimp (Fenneropenaeus chinensis) by suppression subtractive hybridisation (SSH) coupled with the SMART cDNA method. The isolated 944 bp full-length cDNA contains a single 666bp open reading frame (ORF) encoding a putative OMT protein of 221 amino acids. The predicted protein has a molecular weight of 24,572.06 Da and a pI of 5.27 as well as ten phosphorylation sites. Northern blot and in situ hybridisation analyses demonstrated that the OMT transcripts were constitutively expressed in tissue of shrimp challenged by bacterial infection and in unchallenged shrimp tissue. Constitutive OMT transcript was found in areas such as haemocytes, heart, hepatopancreas, stomach, gill, intestine and ovary. However, the OMT transcripts were upregulated in hepatopancreas and stomach in challenged shrimp.

Animal Structures↗

PubMed2006

Effect of sulfide on the immune response and susceptibility to Vibrio alginolyticus in the kuruma shrimp Marsupenaeus japonicus.

Kuruma shrimp Marsupenaeus japonicus held in 34 per thousand seawater were injected with tryptic soy broth (TSB)-grown Vibrio alginolyticus (2.7x10(6)cfu shrimp(-1)), and then placed in water containing concentrations of sulfide at 0 (control), 51, 106, 528 and 1050microgl(-1), respectively. After 12-144h, mortality of V. alginolyticus-injected shrimp exposed to 528 and 1102microgl(-1) sulfide was significantly higher than that of shrimp exposed to 51microgl(-1) sulfide and the control solution. In another experiment, M. japonicus which had been exposed to 0, 56, 112, 525 and 1076microgl(-1) sulfide for 6, 12, 24 and 48h were examined for immune parameters, and phagocytic activity and clearance efficiency of V. alginolyticus. Sulfide concentrations at 525microgl(-1) or greater for 12h resulted in decreased total haemocyte count (THC) and phenoloxidase activity, phagocytic activity and bacterial clearance efficiency, whereas a sulfide concentration at 1076microgl(-1) for 24h caused a significant increase in respiratory burst and superoxide dismutase activity of M. japonicus. It is concluded that concentrations of sulfide at 528microgl(-1) or greater increased the susceptibility of M. japonicus against V. alginolyticus infection by a depression in immune ability. The increased production of superoxide anion by M. japonicus exposed to 525microgl(-1) sulfide or greater was considered to be cytotoxic to the host.

Animals↗

PubMed2006

A transglutaminase from Chinese shrimp (Fenneropenaeus chinensis), full-length cDNA cloning, tissue localization and expression profile after challenge.

Transglutaminase can catalyze the cross-linking reaction between soluble clotting protein molecules from the plasma for prevention of excess blood loss from a wound and obstructing micro-organisms from invading the wound in crustaceans. A novel transglutaminase (FcTG) gene was cloned from hemocytes of Chinese shrimp Fenneropenaeus chinensis by 3' and 5' rapid amplification of cDNA ends (RACE) PCR. The full-length cDNA consists of 2972bp, encoding 757 amino acids with a calculated molecular mass of 84.96kDa and a theoretical isoelectric point of 5.61. FcTG contains a typical transglutaminase-like homologue (TGc domain: E-value=1.94e-38). Three catalytic sites (Cys-324, His-391 and Asp-414) are present in this domain. The deduced amino acid sequence of FcTG showed high identity with black tiger shrimp TG, kuruma shrimp TG and crayfish TG. Transcripts of FcTG mRNA were mainly detected in gill, lymphoid organ and hemocytes by RT-PCR. RNA in situ hybridization further confirmed that FcTG was constitutively expressed in hemocytes both in the circulatory system and lymphoid organ. The variation of mRNA transcription level in hemocytes and lymphoid organ following injection of killed bacteria or infection with white spot syndrome virus (WSSV) was quantified by RT-PCR. The up-regulated expression of FcTG in shrimp lymphoid organ following injection of bacteria indicates that it is inducible and might be associated with bacterial challenge.

Amino Acid Sequence↗

PubMed2005

Organization and expression study of the shrimp (Metapenaeus ensis) putative 5-HT receptor: up-regulation in the brain by 5-HT.

To study the mechanism of 5-HT action and the roles of its receptor in the control of reproduction, we have cloned and characterized the gene and the cDNA of a putative 5-HT receptor (5HT1) from the shrimp, Metapenaeus ensis. The 5HT1 gene is intronless in the coding region but consists of two introns in the 5' untranslated region. 5HT1 transcript is 1.8 kb in size and the cDNA consists of an open reading frame of 1230 bp encoding for a protein of 409 amino acid residues. The deduced 5HT1 consists of the characteristic seven hydrophobic transmembrane (TM1-TM7) domains, which share a high amino acid sequence homology to those of the GPCRs. The results from phylogenetic tree analyses indicate that 5HT1 is more closely related to the octopamine/tyramine of the insect and the 5-HT receptors of the vertebrates than to the other G-protein coupled receptors. Although there is no major difference in the tissues' expression pattern of 5-HT in both sexes, the expression level of 5HT1 is much lower in the females than that in the males. 5HT1 expression in the brain and in the eyestalk is also up-regulated in the 5-HT injected shrimp. By in situ hybridization, no difference in the expression pattern of 5-HT was recorded in the eyestalk of males and females, and the pattern of 5HT1 expression in the eyestalk remained unchanged after 5-HT injection. In the eyestalk, 5HT1 transcripts can be detected in neuronal globuli cells and X-organs. The up-regulation of 5HT1 in the eyestalk and brain may be important for the sustained action in the signal transduction pathway or the regulation of other genes. This may represent an auto-regulation of 5HT1 expression in shrimp by 5-HT. To our knowledge, this is the first demonstration of 5-HT stimulation of a putative 5HT1 gene expression in shrimp.

5' Flanking Region↗

PubMed2006

Analysis of veterinary drug residues in shrimp: a multi-class method by liquid chromatography-quadrupole ion trap mass spectrometry.

A liquid chromatography-mass spectrometry (LC-MS) method was developed to screen and confirm veterinary drug residues in raw shrimp meat. This method simultaneously monitors 18 drugs of different classes, including oxytetracycline (OTC), sulfonamides, quinolones, cationic dyes, and toltrazuril sulfone (TOLS). The homogenized shrimp meat is extracted with 5% trichloroacetic acid. The extract is further cleaned using polymer-based SPE. A 50 mm phenyl column separates the analytes, prior to analysis with an ion trap mass spectrometer interfaced with an atmospheric pressure chemical ionization source. This method is able to confirm oxytetracycline residues at 200 ng/g, toltrazuril sulfone at 50 ng/g, sulfaquinoxaline at 20 ng/g, and the other 15 drugs at 10 ng/g or lower levels. An estimate of the level of residues can also be made so that only confirmed samples above action levels will be sent for quantitation. The method is validated with both fortified and incurred samples, using multiple shrimp species as well. This multi-class method can provide a means to simultaneously monitor for a wide range of illegal drug residues in shrimp.

Animals↗

PubMed2006

A TaqMan real-time RT-PCR for quantifying Mourilyan virus infection levels in penaeid shrimp tissues.

A highly sensitive and specific TaqMan real-time quantitative RT-PCR (qRT-PCR) was developed to detect and quantify Mourilyan virus (MoV), a newly described bunya-like virus of penaeid shrimp. The PCR primers and TaqMan probe targeted a 67-nucleotide (nt) sequence in the MoV M RNA segment. Using dilution series of a 849 nt RNA transcribed in vitro from cDNA clone pMoV4.1, the assay could detect down to a single MoV RNA equivalent, reliably detected 10 RNA copies and had a log linear range up to 1 x 10(9) RNA copies. In experimentally infected Penaeus japonicus shrimp, the test was used to quantify increases in MoV loads over time in hemocytes, lymphoid organ and gills. Sequential increases in MoV RNA copy numbers occurred in lymphoid organ and gill tissues collected at 6, 24 and 48 h post-infection. However, RNA copy numbers decreased slightly in hemocytes sampled at 48 h compared to 24 h. The qRT-PCR data correlated well with amplicon yields generated using a conventional RT-nested PCR targeting the same MoV RNA segment. Moreover, histology and in situ hybridisation using shrimp cephalothorax sections identified increases in lymphoid organ spheroid numbers and confirmed that increases in MoV RNA detected in lymphoid organ tissue were due to expansion in the numbers of infected cells. The qRT-PCR assay should find use in high-throughput screening applications to detect MoV in broodstock and postlarvae used for culture or breeding purposes and for tracking changes in infection levels during shrimp grow-out.

Animals↗

PubMed2004

Waste loading in shrimp and fish processing effluents: potential source of hazards to the coastal and nearshore environments.

On average, only 30-40% of the global fishery production is consumed fresh and the rest 60-70% is processed for human consumption and other purposes. Although the proportion of the total fishery production that are processed remained relatively stable over the last decade, the total bulk of processed fishery commodity increased due to the steady increase in the total fishery production. Processing of large bulk of fish, shrimp and other aquatic organisms produces a corresponding large bulk of by-products and wastes. Although recent trend shows that much of these wastes are made into various value added products, considerable quantities are discharged as the processing effluents with large volume of waters used in processing. Reports suggest that fish and shrimp processing effluents are very high in biological oxygen demand (BOD), chemical oxygen demand (COD), total suspended solids (TSS), fat-oil-grease (FOG), pathogenic and other microflora, organic matters and nutrients, etc. Fish and shrimp processing effluents are, therefore, highly likely to produce adverse effects on the receiving coastal and marine environments. Although substantial reduction of the waste loads is possible by applying available simple techniques, this is not in practice in most part of the world due to lack of proper managerial and regulatory approach. The present paper reviews the characteristics of fish and shrimp processing effluents as a potential source of coastal and marine pollution and, using the existing data, analyzes the global production and discharge of waste loads from the processing plants and discusses available options for waste treatment and management.

Fisheries↗

PubMed2004

Residues of selected antibiotics in water and mud from shrimp ponds in mangrove areas in Viet Nam.

In recent years, antibiotics have been used widely in intensive shrimp culture and this may lead to their contamination of the environment. Surveys on residues of trimethoprim (TMP), sulfamethoxazole (SMX), norfloxacin (NFXC) and oxolinic acid (OXLA) in water and mud in shrimp ponds in mangrove areas were conducted in the north as well as in south of Viet Nam in July and August, 2002. The results show that these antibiotics are found in all samples in both shrimp ponds and surrounding canals. The highest concentrations of TMP, SMX, NFXC and OXLA are 1.04, 2.39, 6.06, and 2.50 ppm in water samples; and 734.61, 820.49, 2615.96, 426.31 ppm (based on wet mud weight), respectively. The comparison of antibiotics residues between study sites and types of shrimp ponds will be discussed in this paper. The results also suggest that antibiotics residues may cause harmful effect on ecosystems in the study sites.

Animals↗

PubMed2006

The impact of shrimp farming effluent on bacterial communities in mangrove waters, Ceará, Brazil.

The effects of shrimp farm effluents on bacterial communities in mangroves have been infrequently reported. Classic and molecular biology methods were used to survey bacterial communities from four mangroves systems. Water temperature, salinity, pH, total heterotrophic bacteria and maximum probable numbers of Vibrio spp. were investigated. Genetic profiles of bacterial communities were also characterized by polymerase chain reaction (PCR) amplification of eubacterial and Vibrio 16S rDNA using denaturing gradient gel electrophoresis (DGGE). Highest heterotrophic counts were registered in the mangrove not directly polluted by shrimp farming. The Enterobacteriaceae and Chryseomonas luteola dominated the heterotrophic isolates. Vibrio spp. pathogenic to humans and shrimps were identified. Eubacterial genetic profiles suggest a shared community structure independent of mangrove system. Vibrio genetic profiles were mangrove specific. Neither microbial counts nor genetic profiling revealed a significant decrease in species richness associated with shrimp farm effluent. The complex nature of mangrove ecosystems and their microbial communities is discussed.

Aquaculture↗

PubMed2006

A Toll receptor in shrimp.

Outbreaks of infectious diseases have resulted in high mortality and huge economic losses in penaeid shrimp culture. Interest in understanding shrimp immunity has increased because of its importance in disease control. Here we report cDNA cloning of a Toll receptor from the white shrimp Litopenaeus vannamei. L. vannamei Toll (lToll) is 926 residues, with a putative signal peptide of 19 residues. The protein contains distinct structural/functional motifs of the Toll like receptor (TLR) family, including an extracellular domain containing 16 leucine-rich repeats (LRRs) flanked by cysteine-rich motifs and a cytoplasmic Toll/interleukin-1 receptor (TIR) domain. The lToll TIR domain showed high similarity to Apis mellifer Toll and Drosophila melanogaster Toll, with 59.9% and 54.3% identity, respectively. Reverse-transcription polymerase chain reaction (RT-PCR) analysis showed that lToll was expressed in hemocyte, gill, heart, brain, stomach, intestine, pyloric caecum, muscle, nerve and spermary, with a lower expression level in eyestalk and hepatopancreas. Identification of lToll will help to elucidate the Toll pathway in shrimp innate immunity.

Amino Acid Motifs↗

PubMed2005

Antibiotic resistance in bacteria from shrimp farming in mangrove areas.

Shrimp farming is a sufficiently large and mature industry to have an effective range of antimicrobial agents for most bacterial diseases in shrimp culture. However, at present, there exists great concern over the widespread use of antibiotics in aquaculture, which may result in residue of antibiotics in water and mud, and subsequently, the development of antibiotic resistance in bacteria in the environment. There is limited understanding about the effect of antibiotic residues on bacteria resistance in shrimp farming environment. Therefore, a study was conducted to investigate bacterial resistance to Norfloxacin (NFXC), Oxolinic Acid (OXLA), Trimethoprim (TMP) and Sulfamethoxazole (SMX), which were found in four shrimp farming locations in mangrove areas in Vietnam. Findings indicate that there is a relatively high incidence of bacteria resistance to these antibiotics observed in most of the studied sites, particularly to antibiotics with concentration of 0.1 microg/ml. Yet the relation between concentration of antibiotic residues and incidence of antibiotic resistance is not clearly defined. Among individual antibiotics, the incidence of resistance to TMP and SMX was higher than the others. Identification of bacteria isolated from mud samples by DNA analyzer shows that Bacillus and Vibrio are predominant among bacteria resistant to the antibiotics. The result of the study also indicates that these antibiotics in media degraded more rapidly due to the presence of resistant bacteria.

Animals↗

PubMed2006

Molecular epidemiology of Vibrio nigripulchritudo, a pathogen of cultured penaeid shrimp (Litopenaeus stylirostris) in New Caledonia.

A collection of 57 isolates of Vibrio nigripulchritudo from either diseased or healthy shrimp and from shrimp farms environment was studied in order to gain a better understanding of the epidemiology of this pathogen, notably isolated from two distinct shrimp disease complexes. Molecular typing using two different techniques, arbitrarily primed PCR (AP-PCR) and multi-locus sequence typing (MLST), studied together with experimental pathology data allowed a relevant epidemiological insight into this possibly emerging pathogen. Additionally, results obtained with the two molecular typing techniques were congruent and allowed discriminating the strains associated with the "Summer Syndrome" from strains isolated from other contexts, especially the other shrimp vibriosis "Syndrome 93". These results highlight that the "Summer Syndrome" is most probably caused by an emergent clonal pathogen that therefore deserves surveillance and that AP-PCR can satisfactorily be used for that purpose.

Animals↗

PubMed2006

Evidence of existence of infectious hypodermal and hematopoietic necrosis virus in penaeid shrimp cultured in China.

Infectious hypodermal and hematopoietic necrosis virus is the causative agent of a shrimp disease which causes economic losses on a global scale. A pair of primers, I2814F/I3516R, was designed from the IHHNV genomic sequence (GenBank) that encodes for structural protein corresponding to nucleotides 2814-3516, which amplifies a 703 base pair (bp) region from the virus genome. PCR amplification with the primers generated a product of the expected size from the purified IHHNV DNA of Litopenaeus vannamei and IHHNV-infected penaeid populations but not from the IHHNV-free shrimp, white spot syndrome virus (WSSV) and hepatopancreatic parvovirus (HPV). The PCR amplicon described above was labeled with digoxigenin (DIG)-11-dUTP as a probe used for dot blot hybridization and in situ hybridization test. Under the optimized PCR conditions, the primers were detected by as little as 20 fg of purified IHHNV DNA, which contained only 8.83 x 10(3) copies of IHHNV, a 1000-fold greater than using dot blot hybridization. Sections of histopathology showed eosinophilic intranuclear inclusions (Cowdry type A inclusions or CAIs) in infected tissues while in situ hybridization, cells displayed an intense reaction with the DIG-labeled probe. PCR assay was developed to detect IHHNV in penaeid shrimp and other crustaceans from the rearing ponds of China (March 2001-June 2004). The positive rate was 51.5% (154 out of 299) and 8.3% (2 out of 24) for penaeid shrimp and crab samples, respectively. The survey demonstrated the presence of IHHNV in China.

Animals↗

PubMed2001

Concurrent production of chitin from shrimp shells and fungi.

Crustacean shells constitute the traditional and current commercial source of chitin. Conversely, the control of fungal fermentation processes to produce quality chitin makes fungal mycelia an attractive alternative source. Therefore, the exploitation of both of these sources to produce chitin in a concurrent process should be advantageous and is reported here. Three proteolytic Aspergillus niger (strains 0576, 0307 and 0474) were selected from a screening for protease activity from among 34 zygomycete and deuteromycete strains. When fungi and shrimp shell powder were combined in a single reactor, the release of protease by the fungi facilitated the deproteinization of shrimp-shell powder and the release of hydrolyzed proteins. The hydrolyzed proteins in turn were utilized as a nitrogen source for fungal growth, leading to a lowering of the pH of the fermentation medium, thereby further enhancing the demineralization of the shrimp-shell powder. The shrimp-shell powders and fungal mycelia were separated after fermentation and extracted for chitin with 5% LiCl/DMAc solvent. Chitin isolates from the shells were found to have a protein content of less than 5%, while chitin isolates from the three fungal mycelia strains had protein content in the range of 10-15%. The relative molecular weights as estimated by GPC for all chitin samples were in the 10(5) dalton range. All samples displayed characteristic profiles for chitin in their FTIR and solid-state NMR spectra. All chitin samples evaluated with MTT and Neutral Red assays with three commercial cell lines did not display cytotoxic effects.

Animals↗

PubMed2000

Molecular characterization of an additional shrimp hyperglycemic hormone: cDNA cloning, gene organization, expression and biological assay of recombinant proteins.

The crustacean eyestalk CHH/MIH/GIH neurohormone gene family represents a unique group of neuropeptides identified mainly in crustaceans. In this study, we report the cloning and characterization of the cDNA and the gene encoding the hyperglycemic hormone (MeCHH-B) of the shrimp Metapenaeus ensis. The amino acid sequence of MeCHH-B shows 85% identity to that of MeCHH-A (formerly MeCHH-like neuropeptide). Two separate but identical MeCHH-B genes were identified in the genome of shrimp by library screening and they are located on different CHH gene clusters. The organization of the MeCHH-B gene is identical to other members of the CHH/MIH/GIH neurohormone family. MeCHH-B is expressed at a constant level in the eyestalks of juveniles and mature females. Unlike the MeCHH-A gene, a low level of MeCHH-B transcripts can also be detected in the central nervous system. Interestingly, the expression pattern of MeCHH-B in the eyestalk of vitellogenic females is reversed to that of the MeCHH-A gene. At the middle stage of gonad maturation, a minimum level of MeCHH-B transcript was recorded and a maximum level of MeCHH-A transcript was detected. Recombinant proteins for MeCHH-A and MeCHH-B were produced by a bacterial expression system. The hemolymph glucose level of bilaterally eyestalk-ablated shrimp increased two-fold 1 h after the rCHH injection and then returned to normal after 2 h. The hyperglycemic effect of these fusion proteins is comparable to that of de-stalked shrimp injected with crude extract from a single sinus gland.

Amino Acid Sequence↗

PubMed2000

Serological estimation of prey-protein gut-residence time and quantification of meal size for grass shrimp consuming meiofaunal copepods.

A series of experiments using serological reagents was conducted to examine predation, ingestion and digestion in a model predator-prey system. The harpacticoid copepod Amphiascoides atopus, obtained from mass culture, was used as prey and the grass shrimp, Palaemonetes pugio, as predator. Bulk-gut passage time in P. pugio was measured by visualization of latex beads and ranged from 0.5 to 4 h in starved and continuously-fed grass shrimp. A polyclonal antibody was prepared from crude extracts of A. atopus; cross reactions with P. pugio and three other crustaceans were either negligible or not detected using slide agar-gel-double-immunodiffusion (AGID) and Western blot preparations. The presence of A. atopus antigens was detected with great sensitivity (e.g., seven copepods, 35 µg dry weight, gave positive results) in grass shrimp gut contents even when proteins of other crustacean prey were present. Prey-proteins could be detected for as long as 4 h with AGID and 8 h with Western blot techniques. Individual grass shrimp that were fed A. atopus and consumed from 0 to 98 copepods h(-1) were subjected to Western-blot preparation with chemiluminescence detection and densitometric evaluation. There was a significant curvilinear relationship between protein content and the number of copepod prey ingested. Results suggest that serological techniques can be modified to estimate the mass or abundance of standard-sized prey ingested by field-collected predators.

Journal Article↗