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Neuroendocrine regulation of osmotic and ionic concentrations in the hemolymph of the freshwater shrimp Macrobrachium olfersii (Wiegmann) (Crustacea, Decapoda).

Putative neuroendocrine mediation of osmotic and ionic responses to acute exposure to high salinity medium was investigated in the freshwater shrimp Macrobrachium olfersii (Wiegmann). Homogenates of supra-esophageal or thoracic ganglia, prepared from shrimps exposed to seawater of 21% S for 6 hr, were injected into the abdominal musculature of shrimps previously exposed to freshwater and subsequently exposed to either freshwater or seawater (21% S). Osmotic, sodium, chloride, potassium, magnesium, and calcium concentrations were determined in hemolymph samples removed by intracardiac puncture at time = 0, 1, 3, or 6 hr after homogenate application. Control shrimps were injected with filtered seawater, isosmotic to the hemolymph, and treated similarly. In control shrimps, the osmotic, Na+, Cl-, K+, Mg2+, and Ca2+ concentrations in the hemolymph increased (P less than or equal to 0.05) after 1-hr exposure to seawater. In shrimps injected with homogenates of supra-esophageal ganglion and exposed to seawater, osmotic and ionic concentrations in the hemolymph did not vary with exposure time; in injected shrimps exposed to freshwater, Na+, Cl-, K+, and Mg2+ concentrations decreased (P less than or equal to 0.05) with time. In shrimps injected with homogenates of thoracic ganglion and exposed to seawater, hemolymph osmotic, K+, and Mg2+ concentrations increased (P less than or equal to 0.05); Na+, Cl-, and Ca2+ concentrations remained unchanged. In injected shrimps exposed to freshwater, hemolymph osmotic concentration alone increased (P less than or equal to 0.05) after 1 hr, all other ionic concentrations remaining unchanged. These data suggest that neurofactors apparently located within the ganglia of the central nervous system of M. olfersii may alter the apparent ionic permeabilities of this shrimp, depending on the salinity characteristics of the external medium. The data support the notion that invasion of the freshwater biotope by estuarine crustaceans has necessitated the evolution of specific physiological mechanisms capable of compensating for the osmotic dilution and ion loss typically encountered by such organisms.

Analysis of Variance↗

Peroxinectin, a cell adhesive protein associated with the proPO system from the black tiger shrimp, Penaeus monodon.

Upon activation of the prophenoloxidase activating system in the shrimp, Penaeus monodon, a cell adhesion activity in the haemolymph is generated. A cell adhesion assay showed that a high number of granular cells (60%) adhered to coverslips coated with a shrimp haemocyte lysate supernatant, whereas a very low number of cells adhered to coverslips coated with bovine serum albumin. Inhibition of adhesion by an antiserum against crayfish peroxinectin, a cell adhesion protein, revealed that the cell adhesion activity detected in shrimp haemocyte lysate supernatant might result from a peroxinectin-like molecule in shrimp. A cDNA clone encoding shrimp peroxinectin was isolated, which had an open reading frame of 2337 nucleotides, with a polyadenylation sequence and a poly A tail. It encodes a protein of 778 amino acids including a 20 amino acid signal peptide. The mature protein (758 amino acids) has a predicted molecular mass of 84.8kDa and an estimated pI of 9.0. Two putative integrin binding motifs, RGD (Arg-Gly-Asp) and KGD (Lys-Gly-Asp), were found in shrimp peroxinectin. Sequence comparison shows that the shrimp protein is similar to crayfish peroxinectin (69%) and to various peroxidases and putative peroxidases from invertebrates and vertebrates. The shrimp peroxinectin cDNA also shows similarity (51%) to both Drosophila peroxinectin-related protein (AAF78217) and peroxidasin (S46224), an extracellular matrix protein combining an active peroxidase domain as well as immunoglobulin domains, leucine rich repeats and procollagen-like motif. However, the sequence similarity to both Drosophila molecules are mostly within the peroxidase domain. Northern blot analysis, using a non-peroxidase region in peroxinectin as a probe, revealed that peroxinectin is constitutively expressed in shrimp haemocyte and was reduced significantly in shrimp injected with a beta-1,3-glucan, laminarin, to mimic an infection with a fungus.

Amino Acid Sequence↗

Toxicity of the mosquito control pesticide Scourge to adult and larval grass shrimp (Palaemonetes pugio).

This study investigated the toxicity of various concentrations of technical resmethrin and Scourge on adult and larval Palaemonetes pugio, a common grass shrimp species. Two types of tests were conducted for each of the resmethrin formulations using adult and larval grass shrimp life stages, a 96-h static renewal aqueous test without sediment, and a 24-h static nonrenewal aqueous test with sediment. For resmethrin, the 96-h aqueous LC50 value for adult shrimp was 0.53 microg/L (95% confidence interval (CI): 0.46-0.60 microg/L), and for larval shrimp was 0.35 microg/L (95% CI: 0.28-0.42 microg/L). In the presence of sediment, technical resmethrin produced a 24-h LC50 value for adult shrimp of 5.44 microg/L (95% CI: 4.52-6.55 microg/L), and for larval shrimp of 2.15 microg/L (95% CI: 1.35-3.43 microg/L). For Scourge, the 96-h aqueous LC50 for adult shrimp was 2.08 microg/L (95% CI: 1.70-2.54 microg/L), and for larval shrimp was 0.36 microg/L (95% CI: 0.24-0.55 microg/L). The 24-h sediment test yielded an LC50 value of 16.12 microg/L (95% CI: 14.79-17.57 microg/L) for adult shrimp, and 14.16 microg/L (95% CI: 12.21-16.43 microg/L) for larvae. Adjusted LC50 values to reflect the 18% resmethrin concentration in Scourge are 0.37 microg/L (adult), 0.07 microg/L (larvae) for the 96-h aqueous test, and 2.90 microg/L (adult), 2.6 microg/L (larvae) for the 24-h sediment test. Larval grass shrimp were more sensitive to technical resmethrin and Scourge than the adult life stage. The results also demonstrate that synergized resmethrin is more toxic to P. pugio than the nonsynergized form, and that the presence of sediment decreases the toxicity of both resmethrin and Scourge.

Animals↗

Quantifying the vitamin K requirement of juvenile marine shrimp (Penaeus monodon) with menadione.

A feeding trial was conducted to estimate the optimum dietary vitamin K requirement for juvenile marine shrimp, Penaeus monodon. Purified diets with eight levels (0, 5, 10, 20, 50, 80, 160 and 320 mg/kg) of supplemental menadione were fed to P. monodon (mean weight 1.33 +/- 0.05 g) for 12 wk. Each diet was fed to three replicate groups of shrimp. Shrimp fed diets supplemented with vitamin K (5-320 mg/kg) had significantly (P < 0.05) higher weight gain and feed efficiency than those fed the unsupplemented control diet. Shrimp fed diets supplemented with > or = 50 mg/kg vitamin K had higher protein efficiency ratios than shrimp fed the control diet. Calcium deposition in shrimp generally increased as dietary vitamin K supplementation increased. Vitamin K-dependent carboxylase activity was highest in shrimp fed the control diet, followed by shrimp fed 5-160 mg menadione/kg diet, and lowest in those fed 320 mg menadione/kg diet. Vitamin K-dependent protein precursor concentrations were high in shrimp fed 0-10 mg menadione/kg diet, lower in those fed 20 mg/kg, and lowest in shrimp fed > or = 50 mg/kg diet. Protein efficiency ratio and vitamin K-dependent protein precursor concentration analyzed by broken-line regression indicated that the adequate dietary vitamin K concentration in growing P. monodon is 30-40 mg/kg diet.

Animal Feed↗

Dietary copper requirement of juvenile grass shrimp, Penaeus monodon, and effects on non-specific immune responses.

An 8-week feeding trial was conducted to determine the dietary copper (Cu) requirement and its effect on the non-specific immune responses of juvenile grass shrimp, Penaeus monodon. Purified diets with seven levels (0, 10, 20, 30, 40, 80, 160 mg Cu kg diet(-1) of supplemental Cu were fed to P. monodon (mean initial weight 0.29 +/- 0.004 g). Each diet was fed to three replicate groups of shrimp. The rearing water contained 1.53 microg Cu 1(-1). Shrimp fed diets supplemented with 10 and 20 mg Cu kg diet(-1) had significantly (P < 0.01) greater weight gain, feed efficiency (FE) and protein efficiency ratio (PER) than those fed the unsupplemented control diet and diets supplemented with > or = 40 mg Cu kg diet(-1). Whole body Cu concentration in shrimp generally increased as dietary Cu supplementation increased. Total haemocyte count (THC) was higher in shrimp fed diets supplemented with 10-30 mg Cu kg diet(-1) than shrimp fed the unsupplemented control diet and diets supplemented with > or = 40 mg Cu kg diet(-1). Intracellular superoxide anion (O2-) production ratios were significantly higher in shrimp fed diets supplemented with 10-30 mg Cu kg diet(-1) than shrimp fed the diet supplemented with 160 mg Cu kg diet(-1). Analysis by polynomial regression of weight gain percent, FE and by linear regression of the whole-body Cu retention of shrimp indicated that the adequate dietary Cu concentration in growing P. monodon is about 15-21 mg Cu kg diet (-1). The immune indicators suggest that an adequate dietary Cu concentration for non-specific immune responses in P. monodon is about 10-30 mg Cu kg diet(-1).

Animal Feed↗

Differences in the susceptibility of American white shrimp larval substages (Litopenaeus vannamei) to four vibrio species.

The rapid expansion of commercial culture of penaeid shrimp is threatened by Vibrio diseases affecting survival and growth. These opportunistic microorganisms are considered part of the normal ecosystem of penaeid shrimp and cause diseases only under conditions that favor them over the host. Shrimp larvae show different susceptibility to these pathogenic agents. In the present work, we report on a comparative study of the susceptibility of all American white shrimp (Litopenaeus vannamei) larval substages to four potentially pathogenic Vibrio species (V. harveyi, V. parahaemolyticus, V. alginolyticus, and V. penaeicida). Strains of these bacterial species were used to infect nauplii, protozoea I-III, mysis I-III, and postlarvae 1 by immersion challenge at 10(3), 10(5), or 10(7) cfu mL(-1) for 30 min. V. alginolyticus infection had no significant effect on survival rate, compared to control, in all shrimp larvae and at all doses tested. Shrimp larvae infected with V. alginolyticus showed a high survival rate compared to other Vibrio species at the three dose levels. V. penaeicida produced a significant mortality effect (P < 0.01) in all shrimp substages and only in postlarvae 1 at low infection dose (10(3) cfu mL(-1)). V. harveyi and V. parahaemolyticus induced significant mortality rates (P < 0.01) only at high doses in shrimp larvae. In summary, shrimp larvae demonstrated an age susceptibility that depends on the Vibrio species and dose level.

Animals↗

Differences in prey capture in grass shrimp, Palaemonetes pugio, collected along an environmental impact gradient.

The waterways and associated salt marshes along the western border of Staten Island, New York (Arthur Kill) have long been under environmental duress. Environmental threats include industrial and municipal discharges, oil spills, and possible leachate from landfills. These impacts are compounded due to the low flushing of this body of water. Grass shrimp, Palaemonetes pugio, inhabiting the Arthur Kill are, therefore, potentially at risk of exposure to metal and organic pollutants. Successful prey capture (of live brine shrimp, Artemia franciscana) was used to compare the relative health of shrimp collected from three sites along an environmental impact gradient. Study sites included a relatively unimpacted harbor (Great Kills Harbor, GK) and two creeks adjoining the Arthur Kill (Nassau Creek, NC, and Richmond Creek, RC). Shrimp originating from GK exhibited a rate of prey capture (6.3 prey h(-1)) that was about two times greater (p < 0.05) than that of shrimp originating from a creek behind a series of landfills (RC, 3.2 prey h(-1)). The rate of prey capture for shrimp collected from a creek impacted by historic smelting activities (NC) was intermediate (5.4 prey h(-1)). Laboratory studies with shrimp from a pristine site (Tuckerton, NJ) exposed to RC conditions (i.e., sediment and water) for eight weeks indicate that reduced prey capture can be induced in healthy shrimp. Finally, video analysis suggests that reduced prey capture in RC shrimp may not be the result of less effort, but rather the combination of (1) 80% fewer (p < 0.05) prey being captured with a lunge type of attack and (2) a greater reliance (p < 0.05) on a less efficient grab type of foraging behavior (64% success rate for RC versus 87% success rate for GK; p = 0.058). These results indicate that sublethal toxicity in environmentally impacted populations can occur and that prey capture may be used to assay the relative health of field specimens. Additionally, impaired prey capture may have important implications for the energy flow within impacted environments.

Animals↗

The environmental impact of shrimp aquaculture: causes, effects, and mitigating alternatives.

Attracted by the demand for shrimp in the developed countries, shrimp aquaculture has expanded rapidly, mainly in the subtropical and tropical lowlands of America and Asia. This work provides a global review and viewpoint on the environmental impacts of shrimp aquaculture, considering the causes and effects of the siting and operation of shrimp ponds and abandonment of farm facilities. Additionally, mitigating alternatives are discussed. To date, approximately 1-1.5 million ha of coastal lowlands have been converted into shrimp ponds, comprising mainly salt flats, mangrove areas, marshes, and agricultural lands. The impact of shrimp farming of most concern is the destruction of mangroves and salt marshes for pond construction. Compatibility with other users, the presence of buffer zones, maintaining an acceptable balance between mangroves and shrimp pond area, improved pond design, reduction of water exchange, and an improved residence time of water, size and capacity to assimilate effluents of the water body, are examples of ways to mitigate the adverse effects. The use of mangroves and halophytes as biofilters of shrimp pond effluents offers an attractive tool for reducing the impact in those regions where mangrove wetlands and appropriate conditions for halophyte plantations exist. Healthy seed supply, good feed with the use of prophylactic agents (including probiotics), good water quality, and lower stocking densities are examples of actions suggested to control disease in shrimp farming. Finally, in the context of integrated management, research priorities are suggested.

Agrochemicals↗

Risk assessment of heavy metal contamination in shrimp farming in Mai Po Nature Reserve, Hong Kong.

An ecological survey was carried out to determine the sediment concentrations of nutrients and heavy metals and bioaccumulation of heavy metals in fish and shrimp including tilapia (Oreochromis mossambicus x O. nilotica), grey mullet (Mugil cephalus), gei wai shrimp (Metapenaeus ensis) and caridean shrimp (Macrobrachium nipponensis) in the traditional tidal shrimp ponds (gei wais) of Mai Po Nature Reserve, Hong Kong. The sediments collected from the landward sites contained higher nutrient contents, as well as zinc (Zn), chromium (Cr), copper (Cu), nickel (Ni) and cadmium (Cd) than those collected from the seaward sites, but vice versa for lead (Pb) and mercury (Hg). However, the concentrations of all metals were exceptionally high in the two sites located outside the reserve, suggesting that waters from Deep Bay might be the possible source of metal contamination affecting the reserve. All metals studied seemed to accumulate in the viscera of fish. Body size was the determining factor for the accumulation of heavy metals in caridean shrimp and gei wai shrimp but not fish. Concentrations of the metals studied in tissues of grey mullet and gei wai shrimp were found to be safe for human consumption. Concentrations of Cr in tilapia whole body (0.68-1.10 mg kg(-1) wet weight) were close to or over the guideline value of 1 mg kg(-1) set by the Food Adulteration (Metallic Contamination) Regulations of Hong Kong. Tilapia flesh and small caridean shrimp collected from gei wais were contaminated by Cr and Pb but still fit for human consumption. Caution is required if large caridean shrimp is to be consumed in large amounts continuously because the concentration of Pb exceeded the maximum permitted concentration (6 mg kg(-1)). The rather high Cr concentrations in tilapia whole body should not be overlooked as the fish will serve as a food source for migratory birds visiting the site.

Animals↗

Provocation-challenge studies in shrimp-sensitive individuals.

Thirty individuals with history of immediate, objective, adverse reactions after shrimp ingestion underwent double-blind, placebo-controlled shrimp-food challenges. All individuals who did not exhibit a positive response (reproduction of objective symptoms) were administered an open challenge of 16 whole cooked shrimp. Positive challenge responses occurred in 9/30 subjects (30%); six of these subjects experienced a positive response during the double-blind phase. Of the 21 remaining subjects, 12 experienced generalized pruritus as their only symptom, whereas nine subjects had completely negative challenge responses. All placebo challenges were negative. Although a positive skin test was strongly associated with challenge symptoms (p less than 0.001), the shrimp prick skin test titration end points were not different among the challenge groups. The serum shrimp RAST percent was significantly higher in the positive challenge group (p less than 0.02). Mean levels of shrimp-specific serum IgG, IgA, and IgM levels were not different among the challenge groups. Although no single immunologic variable could be consistently used to identify subjects more likely to exhibit a positive challenge response, the composite of a positive shrimp prick skin test and elevated serum shrimp-specific IgE (RAST percent label bound greater than 11%) demonstrated a correct predictive value of 87% in this group of shrimp-sensitive subjects.

Allergens↗

A freshwater shrimp (Paratya compressa improvisa) as a sensitive test organism to pesticides.

The susceptibility of 2-week-old individuals of the freshwater shrimp, Paratya compressa improvisa, to five kinds of insecticide and five kinds of herbicide was examined in comparison with that of two species of Cladocera, Daphnia magna and Moina macrocopa. The shrimp was especially sensitive to two organophosphorus insecticides. The 48-h LC50 values for fenitrothion and fenthion to the shrimp were 1.15 and 1.04 microg litre(-1) (mean value, n=2), in contrast with 37.8 and 35.3 microg litre(-1) in the case of M. macrocopa, and more than 50 microg litre(-1) with D. magna. The shrimp also showed the higher susceptibility to other insecticides, diazinon, carbaryl (NAC) and BPMC, apart from D. magna to diazinon and NAC. The shrimp also showed higher susceptibility to herbicides. The 48-h LC50 values of CNP, benthiocarb, oxadiazon, butachlor, and symetryne to the shrimp were two to eight times lower than those of two species of Cladocera, except for the LC50 value of oxadiazon to M. macrocopa, which was very slightly higher. However, the shrimp showed a somewhat lower susceptibility to heavy metals than the two species of Cladocera, especially to copper, and to cadmium and zinc in comparison with D. magna. A bioassay using the shrimp with river water, collected from the river adjacent to the paddy field, showed clearly the high mortality of the shrimp following the aerial spraying with pesticides.

Journal Article↗

Molecular cloning and characterisation of prophenoloxidase from haemocytes of the white shrimp, Litopenaeus vannamei.

cDNA encoding prophenoloxidase (proPO) of the white shrimp Litopenaeus vannamei was obtained from haemocytes by a reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA using oligonucleotide primers based on the proPO sequence of tiger shrimp Penaeus monodon, freshwater crayfish Pacifastacus leniusculus, green tiger shrimp Penaeus semisulcatus (accession no.: AF521949) and kuruma shrimp Marsupenaeus japonicus (accession no.: AB0733223). proPO of L. vannamei was constitutively expressed. The 2471-bp cDNA contained an open reading frame (ORF) of 2058 bp, a 96-bp 5'-untranslated region, and a 317-bp 3'-untranslated region containing the poly A tail. The molecular mass of the deduced amino acid sequence (686 amino acids) was 78.1 kDa with an estimated pI of 6.02. It contained putative copper binding sites, a complement-like motif (GCGWPQHM), a proteolytic activation site, and a conserved C-terminal region common to all known proPOs. However, no signal peptide sequence was detected in white shrimp proPO. Comparison of amino acid sequences showed that white shrimp proPO is more closely related to the proPO of another penaeid than to that of a freshwater crayfish. White shrimp proPO mRNA was synthesized in haemocytes and not in the hepatopancreas or muscle. The activation responses of the proPO of the white shrimp to an exogenous protease (trypsin), a detergent (sodium dodecyl sulphate), and algal and microbial cell wall components (laminarin, sodium alginate, zymosan, and lipopolysaccharide), and its susceptibility to protease inhibitors in vitro resemble the proPO activation system of other crustaceans. These facts suggest that the proPO system in haemocytes of the white shrimp Litopenaeus vannamei serves an important function in non-self recognition and host immune reactions.

Amino Acid Motifs↗

Influence of selected Indian immunostimulant herbs against white spot syndrome virus (WSSV) infection in black tiger shrimp, Penaeus monodon with reference to haematological, biochemical and immunological changes.

Immunostimulants are the substances, which enhance the non-specific defence mechanism and provide resistance against the invading pathogenic micro-organism. In order to increase the immunity of shrimps against the WSSV, the methanolic extracts of five different herbal medicinal plants like Cyanodon dactylon, Aegle marmelos, Tinospora cordifolia, Picrorhiza kurooa and Eclipta alba were selected and mixed thoroughly in equal proportion. The mixed extract was supplemented with various concentrations viz. 100 (A), 200 (B), 400 (C), and 800 (D) mgkg(-1) through artificial diets individually. The prepared diets (A-D) were fed individually to WSSV free healthy shrimp Penaeus monodon with an average weight of 8.0+/-0.5g for 25 days. Control diet (E), devoid of herbal extract was also fed to shrimps simultaneously. After 25 days of feeding experiment, the shrimps were challenged with WSSV, which were isolated and propagated from the infected crustaceans. The shrimps succumbed to death within 7 days when fed on no herbal immunostimulant diet (E). Among the different concentrations of herbal immunostimulant supplemented diets, the shrimps fed on diet D (800mgkg(-1)) significantly (P<0.0001) had more survival (74%) and reduction in the viral load. Also the better performance of haematological, biochemical and immunological parameters was found in the immunostimulant incorporated diets fed shrimps. The present work revealed that the application of herbal immunostimulants will be effective against shrimp viral pathogenesis and they can be recommended for shrimp culture.

Adjuvants, Immunologic↗

Characterization of the shrimp eyestalk cDNA encoding a novel fushi tarazu-factor 1 (FTZ-F1).

To study the role of ecdysone and the ecdysone inducible gene in the regulation of molting and development in crustaceans, we have cloned a cDNA encoding an orphan nuclear receptor family member from the eyestalk of the shrimp Metapenaeus ensis. The size of the cDNA is 4.3 kb with the longest open reading frame (ORF) encoding a protein of 545 amino acid residues. The deduced amino acid sequence of the shrimp cDNA consists of regions that are characteristic of those of the nuclear hormone receptors. It shows a high degree of amino acid sequence identity in the DNA binding domain, ligand binding domain and the FTZ box as compared to those of invertebrates and vertebrates. Unlike the insects Drosophila melanogaster and Bombyx mori, an AF2 transactivation domain was present in the shrimp FTZ-F1. Northern blot analysis using total RNA indicated that the FTZ-F1 mRNA could also be detected in the mature ovary. Northern blot analysis and RT-PCR analysis showed that the shrimp FTZ-F1 transcripts could be detected in the ovary, newly hatched nauplius, testis, eyestalk and epidermis of the adult shrimp. Although the cDNA clone was isolated from the eyestalk library, the shrimp FTZ-F1 appeared to express most abundantly in the mature oocytes. The presence of abundant FTZ-F1 specific maternal message in the late vitellogenic ovary and early nauplius indicates that it may be important for the early embryonic and larval development of the shrimp. Interestingly, shrimp FTZ-F1 can also be found in testis of the male shrimp. The presence of FTZ-F1 in other tissues such as epidermis suggests that it may also be involved in other physiological processes such as molting.

Amino Acid Sequence↗

Chemicals and biological products used in south-east Asian shrimp farming, and their potential impact on the environment--a review.

A wide variety of chemicals and biological products are used to treat the water and sediment of ponds in semi-intensive and intensive south-east Asian shrimp farming. These products are also often used in shrimp hatcheries and to disinfect equipment for shrimp pond management. In spite of the size and importance of the shrimp farming industry in several south-east Asian countries, documentation of the quality and quantity of chemicals and biological products used during farming is scarce. This paper is a compilation of the literature available on substances used in shrimp farming, and the possible environmental effects of these products are analysed to the extent allowed by the limited information. The role of shrimp farm managers, the chemical industry, governments, inter-governmental organisations and scientists in the development of a sustainable practice is discussed. It is concluded that shrimp farmers should reduce the use of chemicals and biological products because of the risks to the environment, human health and to production, and also, because many chemicals and biological products used in pond management have not been scientifically shown to have a positive effect on production. Clearly, the use of some chemicals, i.e. certain antibiotics, poses a risk of danger towards human health. Some chemicals used in shrimp farming, such as organotin compounds, copper compounds, and other compounds with a high affinity to sediments leave persistent, toxic residues, and are likely to have a negative impact on the environment. However, to assess the reality of these risks, substantial new information about the quantity of chemicals used in marine south-east Asian shrimp farming is needed.

Animals↗

Growth and enterotoxin production of Staphylococcus aureus in shrimp.

Strains of Staphylococcus aureus isolated from shrimp were examined for phage pattern and enterotoxin production; 63% of the strains isolated from North Sea shrimp were typable with the International and additional set of phages, as were 38% of the strains isolated from South-East Asian shrimp. Staphylococcal enterotoxin(s) (SE) were produced by 48% and 35% of strains isolated from North Sea and South-East Asian shrimp respectively. Growth and enterotoxin production by S. aureus in shrimp was examined in storage experiments at 22 degrees C. S. aureus increased by 1-2 log units in 24 h when the organism was only a minor part of the total microflora of shrimp. When S. aureus was an equivalent part of the total flora its numbers increased by 3-4 log units in 24 h. Enterotoxins A and B became detectable when the number of S. aureus exceeded 10(7) per g in aseptically peeled shrimp. Results indicate that S. aureus is able to produce enterotoxin in shrimp, but its production depends upon a number of factors, including the relationship between S. aureus and competitive micro-organisms. It is concluded that the presence of S. aureus on commercially produced shrimp represents a potential hazard to health.

Animals↗

Immunoglobulin E antibody reactivity to the major shrimp allergen, tropomyosin, in unexposed Orthodox Jews.

BACKGROUND: Assessment of allergic (IgE antibody-mediated) reactions to foods may become complicated by cross-reactivity that can occur among certain food families and between foods and seemingly unrelated allergens. OBJECTIVE: The allergenic properties of tropomyosin (muscle-derived protein) have been recently demonstrated in invertebrates such as cockroaches, dust mites, and shrimp. In view of a possible cross-reactivity between food allergens and related allergens from animal sources, we designed a study to assess IgE antibody reactivity to the major shrimp allergen, Pen a 1, in an unexposed population of Orthodox Jews, who observe Kosher dietary laws that prohibit eating shellfish. METHODS: Nine subjects, who reacted positively by skin tests to shrimp (Penaeus setiferous), were selected for the study. Subjects (two females, seven males) ranged in age from 14 to 32 years (mean 20.4). All subjects were strictly observant of Jewish tradition and had no prior exposure to seafood (regarded as a non-Kosher food). Serum was obtained from all the subjects and tested for IgE antibody reactivity to shrimp and dust mite. RESULTS: All subjects reported symptoms of perennial allergic rhinitis, five had history of asthma, atopic dermatitis, and/or sinusitis. All had positive skin prick tests to shrimp and house dust mite (HDM) (Dermatophagoides farinae, D. pteronyssinus, or both); 2/7 subjects were positive to cockroach mix (Blattella germanica and Periplaneta americana). Sera of 4/9 subjects demonstrated specific IgE antibodies by RAST to shrimp (7.0-20.0%), 3/9 to Pen a 1 (6.3-24.1%), and 3/9 to shrimp or Pen a 1 by immunoblot. IgE binding to Pen a 1 was inhibited with either mite or cockroach extracts as demonstrated by RAST and/or immunoblot inhibition analysis. CONCLUSIONS: These studies indicate that IgE antibody reactivity to a major food allergen, shrimp, can occur in an unexposed population of individuals; some subjects allergic to HDM and/or cockroach show substantial IgE antibody reactivity to the major shrimp allergen Pen a 1 (tropomyosin). Based on inhibition with cockroach and/or dust mite extracts, this reactivity appears to be due to cross-reacting tropomyosins.

Allergens↗

A non-destructive method based on the polymerase chain reaction for detection of hepatopancreatic parvovirus (HPV) of penaeid shrimp.

Current methods to detect hepatopancreatic parvovirus (HPV) infection of penaeid shrimp depend on invasive techniques that require dissecting the organs infected by this virus. However, sacrificing valuable stocks in order to determine their HPV status can be a drawback in the case of breeding programs. A method was developed for HPV detection by applying a polymerase chain reaction (PCR) assay to fecal samples collected from live HPV-infected shrimp Penaeus chinensis. A pair of PCR primers, 1120F/1120R, which amplify a 592 base pair (bp) region from the virus genome, was designed from previously known HPV sequence information (HPV clone HPV8). PCR amplification with these primers generated a product of the expected size directly from the crude feces of HPV-infected shrimp but not from the feces of specific pathogen-free (SPF) shrimp. The HPV origin of the amplified product was validated by means of an in situ hybridization assay where the product of the amplification, labeled with digoxigenin (DIG)-11-dUTP, showed an intense reaction within hepatopancreatic cells displaying characteristic HPV lesions on HPV-infected shrimp. No reaction to this probe was observed when reacted in situ with sections of the hepatopancreas of SPF specimens or to sections of shrimp infected by the infectious hypodermal and hematopoietic necrosis virus (IHHNV), another parvovirus of penaeid shrimp. These primers were tested for specificity against homologous and nonhomologous viruses and no product was amplified. A fragment of the expected size was obtained only when purified HPV or purified HPV8 plasmid was used as template DNA. Under optimized conditions, these primers detected as little as 1 fg of purified HPV8 plasmid DNA, equivalent to approximately 300 HPV particles. Analysis of fecal samples by PCR may prove useful for non-lethal screening of valuable shrimp of unknown HPV status. This same strategy also might be used for detection of other enteric viruses that infect penaeid shrimp.

Animals↗