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Nitrogen and phosphorus cycling in shrimp ponds and the measures for sustainable management.

Six ponds of age 3 were selected 45 km north from Suzhou in the Tailake region, and research conducted on nitrogen and phosphorus cycling in P. vannanmei (Penaeus vannanme) ponds and M. nipponense (Macrobrachium nipponense) hatchery ponds under normal management. Two treatments each had three replications. The results confirmed that feed was the major path of nitrogen and phosphorus input, each accounted for 61.24% (193.81 kg ha(-1)) and 81.08% (45.20 kg ha(-1)) of the total nitrogen and phosphorus input for P. vannanme ponds; the values for M. nipponense ponds were 43.93% (86.31 kg ha(-1)) and 57.67% (14.61 kg ha(-1)), respectively. Water pumped into ponds contributed on average 83.57 kg ha(-1) nitrogen and 8.48 kg ha(-1) phosphorus for P. vannanmei ponds, and 87.48 kg ha(-1) nitrogen and 7.00 kg ha(-1) phosphorus for M. nipponense hatchery ponds. Shrimp harvest recovered 102.81 kg ha(-1) nitrogen (32.94% of the total nitrogen input) and 7.94 kg ha(-1) phosphorus (14.23% of the total phosphorus input) for P. vannanme ponds; and 43.94 kg ha(-1) nitrogen and 4.46 kg ha(-1) phosphorus for M. nipponense hatchery ponds. The sum of nitrogen losses through volatilization, denitrification and sedimentation was 173.62 and 122.39 kg ha(-1), 54.86% and 62.29% of the total nitrogen input for P. vannanme ponds and M. nipponense hatchery ponds, respectively. Sediment accumulated 41.46 and 14.63 kg ha(-1) phosphorus, 74.37% and 64.85% of the total phosphorus input for P. vannanm ponds and M. nipponense hatchery ponds. Draining and seeping caused 40.06 kg ha(-1) nitrogen (12.66% of total nitrogen input) and 6.36 kg ha(-1) phosphorus (11.40% of total phosphorus input) loss to the surrounding water from P. vannanme ponds in 114 days; 30.14 kg ha(-1) nitrogen (15.34% of the total input) and 4.45 kg ha(-1) phosphorus (17.57% of the total input) to channel water from M. nipponense hatchery ponds in 87 days, respectively. Countermeasures for sustainable pond management include improving feeds and feeding, sediment treatments, machine aerating, chemicals with no pollution, and integrated fish-shrimp cultivation. Management of water resources for pond and methods to reduce nitrogen and phosphorus loading into surrounding water from drainage are elucidated.

Aquaculture↗

A novel eye in 'eyeless' shrimp from hydrothermal vents of the Mid-Atlantic Ridge.

Rimicaris exoculata is a shrimp that swarms over high-temperature (350 degrees C) sulphide chimneys at Mid-Atlantic Ridge hydrothermal fields (3,600 m). This shrimp lacks an externally differentiated eye, having instead a pair of large organs within the cephalothorax immediately beneath the dorsal surface of the transparent carapace, connected by large nerve tracts to the supraesophageal ganglion. These organs contain a visual pigment with an absorption spectrum characteristic of rhodopsin. Ultrastructural evidence for degraded rhabdomeral material suggests the presence of photoreceptors. No image-forming optics are associated with the organs. We interpret these organs as being eyes adapted for detection of low-level illumination and suggest that they evolved in response to a source of radiation associated with the environment of hydrothermal vents.

Animals↗

Snapping shrimp make flashing bubbles.

Snapping shrimp produce a loud crackling noise that is intense enough to disturb underwater communication. This sound originates from the violent collapse of a large cavitation bubble generated under the tensile forces of a high-velocity water jet formed when the shrimp's snapper-claw snaps shut (Fig. 1). Here we show that a short, intense flash of light is emitted as the bubble collapses, indicating that extreme pressures and temperatures of at least 5,000 K (ref. 4) must exist inside the bubble at the point of collapse. We have dubbed this phenomenon 'shrimpoluminescence' - the first observation, to our knowledge, of this mode of light production in any animal - because of its apparent similarity to sonoluminescence, the light emission from a bubble periodically driven by ultrasound.

Animals↗

Biomechanics: deadly strike mechanism of a mantis shrimp.

Stomatopods (mantis shrimp) are well known for the feeding appendages they use to smash shells and impale fish. Here we show that the peacock mantis shrimp (Odontodactylus scyllarus) generates an extremely fast strike that requires major energy storage and release, which we explain in terms of a saddle-shaped exoskeletal spring mechanism. High-speed images reveal the formation and collapse of vapour bubbles next to the prey due to swift movement of the appendage towards it, indicating that O. scyllarus may use destructive cavitation forces to damage its prey.

Animals↗

A new class (penaeidin class 4) of antimicrobial peptides from the Atlantic white shrimp (Litopenaeus setiferus) exhibits target specificity and an independent proline-rich-domain function.

A highly pure, chemically defined representative of a new class of antimicrobial peptide from the Atlantic white shrimp (Litopenaeus setiferus), penaeidin class 4 [Pen4-1 (penaeidin class 4 isoform 1)], was produced synthetically. Chemical synthesis was achieved by native ligation from two separate domains yielding a bioactive peptide that reflected the characteristics of native penaeidin. Synthetic Pen4-1 proved to be an effective antimicrobial peptide, particularly against the broad-spectrum pathogen Fusarium oxysporum, exhibiting a complex effect on reproductive growth at inhibitory concentrations resulting in the suppression of spore formation. Pen4-1 exhibits unique features [not previously observed for penaeidins from the Pacific white shrimp (L. vannamei)], including target-species specificity against Gram-positive bacteria, indicating a potential partitioning of antimicrobial function among this family of peptides. The proline-rich domain of penaeidin class 4 alone was an active antimicrobial peptide, having the same target range as the full-length Pen4-1. These findings indicate that the proline-rich domain of penaeidin is sufficient to confer target specificity and that divergence in this domain between classes can result in a gain in antimicrobial function as observed for the proline-rich domain of Pen4-1.

Amino Acid Sequence↗

Control of pathogenic Vibrio spp. by Bacillus subtilis BT23, a possible probiotic treatment for black tiger shrimp Penaeus monodon.

AIMS: The present study evaluated the in vitro and in vivo antagonistic effect of Bacillus against the pathogenic vibrios. METHODS AND RESULTS: Cell-free extracts of Bacillus subtilis BT23 showed greater inhibitory effects against the growth of Vibrio harveyi isolated by agar antagonism assay from Penaeus monodon with black gill disease. The probiotic effect of Bacillus was tested by exposing shrimp to B. subtilis BT23 at a density of 106-108 cfu ml-1 for 6 d before a challenge with V. harveyi at 103-104 cfu ml-1 for 1 h infection. The combined results of long- and short-term probiotic treatment of B. subtilis BT23 showed a 90% reduction in accumulated mortality. CONCLUSIONS: This study reports that pathogenic vibrios were controlled by Bacillus under in vitro and in vivo conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Results indicated that probiotic treatment offers a promising alternative to the use of antibiotics in shrimp aquaculture.

Animals↗

Nutritional availability to rats of selenium in four seafoods: crab (Callinectes sapidus), oyster (Crassostrea virginica), shrimp (Penaeus duorarum) and Baltic herring (Clupea harengus).

1. The present study was conducted to determine the biological availability to rats of the selenium in four high-Se seafoods: crab (Callinectes sapidus), oyster (Crassostrea virginica), shrimp (Penaeus duorarum) and Baltic herring (Clupea harengus). 2. Weanling male rats were fed on a Se-deficient Torula yeast diet for 4 weeks followed by either continued depletion or repletion for 4 weeks with 0.05, 0.1 or 0.2 microgram Se as selenite/g, or 0.1 or 0.2 microgram Se as freeze-dried cooked test food/g. Plasma and liver Se levels or glutathione peroxidase (EC 1.11.1.9; GSH-Px) activities were used as indicators of body Se status. 3. Except for oysters, the biological availability of Se in all these seafoods was close to that of selenite (selenite 100%) when the criterion used was either plasma Se level or plasma GSH-Px activity. 4. By the criterion of increased liver Se level of restored hepatic GSH-Px activity, only herring-Se had a biological availability comparable to that of selenite-Se under all conditions tested, whereas crab-Se and oyster-Se were distinctly inferior in this regard. 5. Increasing the amount of crab-Se, oyster-Se or shrimp-Se supplied in the diet from 0.1 to 0.2 microgram/g changed the apparent availability (%) of Se for hepatic GSH-Px restoration from 38 to 78, 22 to 53 and 57 to 90 respectively. 6. The present study demonstrates that the availability of Se in certain foods is a function of the criterion chosen, the level of Se supplied in the diet, and possibly other unknown interacting dietary factors.

Animal Nutritional Physiological Phenomena↗

Analysis of polyphosphates in fish and shrimps tissues by two different ion chromatography methods: implications on false-negative and -positive findings.

Inorganic polyphosphates (di-, tri- and higher polyphosphates) can be used to treat fish, fish fillets and shrimps in order to improve their water-binding capacity. The practical relevance of this treatment is a significant gain of weight caused by the retention/uptake of water and natural juice into the fish tissues. This practice is legal; however, the use of phosphates has to be declared. The routine control testing of fish for the presence of polyphosphates, produced some results that were difficult to explain. One of the two analytical methods used determined low diphosphate concentrations in a number of untreated samples, while the other ion chromatography (IC) method did not detect them. This initiated a number of investigations: results showed that polyphosphates in fish and shrimps tissue undergo a rapid enzymatic degradation, producing the ubiquitous orthophosphate. This led to the conclusion that sensitive analytical methods are required in order to detect previous polyphosphate treatment of a sample. The polyphosphate concentrations detected by one of the analytical methods could not be explained by the degradation of endogenous high-energy nucleotides like ATP into diphosphate, but by a coeluting compound. Further investigations by LC-MS-MS proved that the substance responsible for the observed peak was inosine monophsosphate (IMP) and not as thought the inorganic diphosphate. The method producing the false-positive result was modified and both methods were ultimately able to detect polyphosphates well separated from natural nucleotides. Polyphosphates could no longer be detected (<0.5 mg kg-1) after modification of the analytical methodology. The relevance of these findings lies in the fact that similar analytical methods are employed in various control laboratories, which might lead to false interpretation of measurements.

Animals↗

Analysis of veterinary drug residues in fish and shrimp composites collected during the Canadian Total Diet Study, 1993-2004.

Thirty shrimp, marine fish, freshwater fish, and canned fish composite samples collected and prepared as part of the Canadian Total Diet Study were analysed for 39 different veterinary drug residues. The analyses were undertaken to obtain baseline data that could be used to estimate the dietary exposure of Canadians to these residues. The most frequently observed residue was AOZ (four out of 30 samples), the metabolite of furazolidone, at a range of 0.50 to 2.0 ng g(-1) wet weight. Other residues detected included enrofloxacin (three samples; 0.3-0.73 ng g(-1)), leucomalachite green (three samples; 0.73-1.2 ng g(-1)), oxolinic acid (two samples; 0.3-4.3 ng g(-1)), AMOZ (the metabolite of furaltadone; one sample; 0.40 ng g(-1)), chloramphenicol (one sample; 0.40 ng g(-1)), and SEM (the metabolite of nitrofurazone; one sample; 0.8 ng g(-1)). The results of this survey indicate that Canadians are exposed to low ng g-1 concentrations of some banned and unapproved veterinary drug residues via the consumption of certain fish and shrimp.

Animals↗

Radiobichemistry of phytodrugs: II. Activity of Altoside (r) and Altozar (R) in the biosynthesis of proteins and RNA in larvae of shrimps in vivo studied by leucine-U-14C and uridine-2-14C.

The present communication concerns the investigation of the effect of various concentrations of Altosid (R), J.H.A., on the biosynthesis of proteins and RNA in cultures of shrimp larvae in vivo measured by the uptake up Leucine, U-14C and Uridine-2-14C. The incorporation of Leucine follows different behaviour in the presence of either 0.01 or 1 ppm concentration of J.H.A.. At any dose Altosid (R) and Altozar (R) decrease the protein synthesis in the larvae of shrimps also at concentrations of 0.01 ppm, the main dose used for their application. In fact in these condition the synthesis of proteins is lower than in the control. For the RNA biosynthesis the uptake of Uridine is much higher in the presence of J.H.A. than in the control. It is suggested then an abnormal s-RNA is formed in the first hours, which does not synthetize proteins.

Animals↗

Evaluation of chicken embryo, brine shrimp, and bacterial bioassays for saxitoxin.

The chicken embryo, brine shrimp (Artemia salina) and selected bacteria (Staphylococcus epidermidis, Micrococcus flavus, and Bacillus subtilis) were evaluated as alternative test systems for the determination of paralytic shellfish poisoning toxicity (saxitoxin). Dose levels ranging from 0.045 to 0.3 microgram were administered to the developing embryo through the air cell at either 0 or 96 h following incubation. Embryos dosed 96 h after incubation were the most sensitive, with 100% mortality at the 0.11-microgram dose level 24 h following exposure. Embryo mortality was 42% at to 0.3-microgram dose level when the toxin was administered at 0 h. Some embryonic malformations were observed in the 0-h treatment group. Brine-shrimp larvae were sensitive to saxitoxin at a dose level of 5 micrograms. A dose response based on mortality was apparent only 48 h after dosing. Limited growth inhibition was observed with the bacteria tested at concentrations between 0.0035 and 0.35 microgram and 0.35 microgram/well.

Animals↗

Multimeric hemoglobin of the Australian brine shrimp Parartemia.

The hemoglobin molecule of the commercially important brine shrimp Artemia sp. has been used extensively as a model for the study of molecular evolution. It consists of nine globin domains joined by short linker sequences, and these domains are believed to have originated through a series of duplications from an original globin gene. In addition, in Artemia, two different polymers of hemoglobin, called C and T, are found which differ by 11.7% at the amino acid level and are believed to have diverged about 60 MYA. This provides a set of data of 18 globin domain sequences that have evolved in the same organism. The pattern of amino acid substitution between these two polymers is unusual, with pairs of equivalent domains displaying differences of up to 2.7-fold in total amino acid substitution. Such differences would reflect a similar range of molecular-clock rates in what appear to be duplicate, structurally equivalent domains. In order to provide a reference outgroup, we sequenced the cDNA for a nine-domain hemoglobin (P) from another genus of brine shrimp, Parartemia zietziana, which differs morphologically and ecologically from Artemia and is endemic to Australia. Parartemia produces only one hundredth the amount of hemoglobin that Artemia produces and does not upregulate production in response to low oxygen partial pressure. Comparison of the globin domains at the amino acid and DNA levels suggests that the Artemia globin T gene has accumulated substitutions differently from the Parartemia P and Artemia C globin genes. We discuss the questions of accelerated evolution after duplication and possible functions for the Parartemia globin.

Amino Acid Substitution↗

Relationship between vitellogenin and vitellin in a marine shrimp (Penaeus semisulcatus) and molecular characterization of vitellogenin complementary DNAs.

The source of yolk proteins in crustacean ovaries has been the subject of controversy for several decades, and both extraovarian and intraovarian synthesized proteins have been implicated. To offer a new insight into the relationship of vitellogenin (VTG) and vitellin (VT), a comparison of extraovarian VTG and ovarian VT of the marine shrimp Penaeus semisulcatus was performed at the protein and cDNA levels. Two cDNAs (7920 and 2068 nucleotides [nt]) were sequenced for VTG from the ovary and one cDNA (7920 nt) was sequenced from the hepatopancreas. VTG cDNA from hepatopancreas was similar to VTG cDNA from ovary. Although a VTG gene was also found in the males, approximately 7.8-kilobase transcripts were only detected in the ovary and hepatopancreas of females. The mRNA expression pattern was related to the stage of ovarian development and to the molt cycle, as determined by real-time polymerase chain reaction assay. VTG and VT apoproteins were composed of two and three major subunits, respectively, as shown by SDS-PAGE. N-terminal sequences of these subunits revealed the presence of a cleavage site at a consensus motif for a subtilisin-like endoprotease in VTG and VT and an additional cleavage site in VT revealed by an unidentified endoprotease. These results indicate that penaeid shrimps constitute a unique model for vitellogenesis, showing intraovarian gene expression and synthesis of yolk protein.

Amino Acid Sequence↗

Hand injuries from shrimp boat winches.

Shrimp boat winch injury to the upper extremity was identified in three patients. The injury occurred when the individual's hand became entrapped in the cable and the upper extremity was drawn into the winch mechanism. This resulted in midshaft fractures of one or both bones of the forearm and amputation of a portion of the fourth and fifth fingers. This injury emphasizes the importance of safeguards on the cable system. However, in the absence of safety regulations in the shrimping industry, more of these injuries can be expected.

Accidents, Occupational↗

Purification and characterization of the infectious hypodermal and haematopoietic necrosis virus of penaeid shrimps.

Infectious hypodermal and haematopoietic necrosis (IHHN) is one of the most important viral diseases of cultured penaeid shrimps and is potentially a limiting factor in the development of farming projects for some species of these shrimps. Although the IHHN agent was recognized early as being viral in origin, attempts to characterize it were inconclusive because of difficulties in obtaining sufficient amounts of purified virions to permit its characterization. Recent improvements of purification procedures have allowed the physicochemical characterization of this virus. Purified IHHNV is a non-enveloped icosahedral particle averaging 22 nm in diameter, exhibiting a mean buoyant density of 1.40 g/ml in CsCl. The genome is a single molecule of ssDNA with an estimated size of 4.1 kb by molecule length measurement in transmission electron microscopy. As determined by SDS-PAGE, the particle contains four polypeptides with Mrs of 74K, 47K, 39K and 37.5K, respectively. From its characteristics, this virus could be a member of the Parvoviridae family.

Animals↗

Characterization of hepatopancreatic parvo-like virus, a second unusual parvovirus pathogenic for penaeid shrimps.

The hepatopancreatic parvo-like virus (HPV) of penaeid shrimp was extracted from infected shrimp tissues, purified and subsequently characterized. The viral particles, icosahedral in shape, are 22 nm in diameter and possess a buoyant density of 1.41 g/ml. They contain ssDNA, of approximately 5 kb in size which encodes a single polypeptide of 54 kDa. On the basis of its general characteristics this pathogenic agent belongs to the Parvoviridae family, but because of two unusual characteristics (capsid protein formed with a single polypeptide and genome structure more closely related to the autonomous parvoviruses rather than the densoviruses), it seems to constitute a novel group in the Parvoviridae family.

Animals↗

Prevalence of Vibrio cholerae with heat-stable enterotoxin (NAG-ST) and cholera toxin genes; restriction fragment length polymorphisms of NAG-ST genes among V. cholerae O serogroups from a major shrimp production area in Thailand.

A total of 148 Vibrio cholerae isolates from a major shrimp production area in Southern Thailand were examined by colony hybridisation for genes encoding heat-stable enterotoxin (NAG-ST) and cholera toxin (CT). Only non-O1 V. cholerae strains were found to harbour NAG-ST (14 of 146) whereas no strains hybridised with the CT probe. NAG-ST-positive V. cholerae non-O1 strains were isolated from shrimp farms situated close to urban areas. Five different O serogroups were found among NAG-ST positive non-O1 strains. Southern blot and restriction endonuclease analysis of NAG-ST-positive strains revealed a high degree of genetic divergence. A total of seven classes of enterotoxin gene patterns were found with HindIII and EcoRI restriction endonucleases. Enterotoxin gene patterns correlated with O-antigen expression in 84% of isolates tested. In combination with other molecular techniques Southern blot analysis with an NAG-ST oligonucleotide probe could be useful for studying the molecular epidemiology of V. cholerae non-O1 strains.

Animals↗

Identification and analysis of gp116 and gp64 structural glycoproteins of yellow head nidovirus of Penaeus monodon shrimp.

Yellow head virus (YHV) is a major agent of disease in farmed penaeid shrimp. YHV virions purified from infected shrimp contain three major structural proteins of molecular mass 116 kDa (gp116), 64 kDa (gp64) and 20 kDa (p20). Two different staining methods indicated that the gp116 and gp64 proteins are glycosylated. Here we report the complete nucleotide sequence of ORF3, which encodes a polypeptide of 1666 amino acids with a calculated molecular mass of 185 713 Da (pI=6.68). Hydropathy analysis of the deduced ORF3 protein sequence identified six potential transmembrane helices and three ectodomains containing multiple sites for potential N-linked and O-linked glycosylation. N-terminal sequence analysis of mature gp116 and gp64 proteins indicated that each was derived from ORF3 by proteolytic cleavage of the polyprotein between residues Ala(228) and Thr(229), and Ala(1127) and Leu(1128), located at the C-terminal side of transmembrane helices 3 and 5, respectively. Comparison with the deduced ORF3 protein sequence of Australian gill-associated virus (GAV) indicated 83 % amino acid identity in gp64 and 71 % identity in gp116, which featured two significant sequence deletions near the N terminus. Database searches revealed no significant homology with other proteins. Recombinant gp64 expressed in E. coli with and without the C-terminal transmembrane region was shown to react with antibody raised against native gp64 purified from virions.

Amino Acid Sequence↗