Protective effect of sodium alginates against bacterial infection in common carp, Cyprinus carpio L.

Abstract. A hot-water extract of Undaria pinnatifida was found to contain polysaccharide(s) which showed immunostimulating activity in fish. In order to identify the effective component(s), the hot-water extract was fractionated and acid-soluble (Fraction I) and acid-insoluble (Fraction II) polysaccharides were evaluated for their potential to enhance protection against Edwardsiella tarda infection in common carp. Intraperitoneal injection of Fraction II in carp 6 and 3 days prior to challenge with E. tarda resulted in a significantly greater survival rate than that of control fish at doses of 10–30mgkg^-1. On the other hand, Fraction I was not effective at any dose. Chemical and physicochemical analyses revealed that Fraction II was sodium alginate with a high degree of purity, with a molecular weight of 45000 and an M/G ratio of 0.8. Some commercial alginates were similarly evaluated for their efficacy. Alginates from Macrocystis pyrifera (MW, 146000-264000; M/G ratio, 0.9–1.0) significantly increased survival rate, but alginates from Lessonia nigrescens (MW, 177000–290000; M/G ratio, 1.3) had little effect. There are indications that the efficacy (protective effect) of alginates has some relationship to their M/G ratios.     

Herpesvirus cyprini: lethality and oncogenicity

Abstract. Cyprinid herpesvirus 1 (CHV) or Herpesvirus cyprini was virulent for carp, Cyprinus carpio L., fry following 1 h immersion in water at 20 °C. Cumulative mortality for carp fry was 86–97% in 2-week-old common carp, 20% in 4-week-old fancy carp, and 0% in both 8-week-old common and fancy carp. The virus did not produce mortality in fry of crucian carp, grass carp or other cyprinids. It was also oncogenic in carp, inducing papillomas to the extent of 55% among both common and fancy carp fry. The neoplasms appeared 5–6 months after carp had been exposed to the virus by immersion and recurred at an incidence of 83% in carp 7·5 months post-desquamation of the tumour. The CHV was reisolated from all moribund fish and from all survivors. It also induced papillomas at an incidence of 13% in adult mirror carp and at 10% in adult fancy carp 5 months after intraperitoneal inoculation of 10⁵ TCID₅₀ ml^-1 fish. The virus was rcisolated only from the ncoplastic tissue and not from internal organs. The neoplasms were normally located on fin, skin or mandible, at the intraperitoneal inoculation site. Specific fluorescence for CHV antigen was frequently detected in the gills, liver, kidneys and intestine of 2-week-old fry from 3 to 21 days following challenge with CHV. It was found in greater concentrations in experimentally induced papillomata on 2-week-old carp fry survivors examined 24 weeks after challenge than in naturally occurring neoplasms.     

Protective properties of egg yolk IgY containing anti-Edwardsiella tarda antibody against paracolo disease in the Japanese eel, Anguilla japonica Temminck & Schlegel

Abstract. Immunized hens are known to contain a high level of immunoglobulin Y (IgY) in their egg yolk. In this study, the present authors obtained anti- Edwardsiella tarda IgY (containing 20% specific IgY, agglutination titre, 1:128) from hens vaccinated by injection with formalin-killed bacterin. The IgY was stable against eel digestive factors, and therefore, was orally administered with viable E. tarda to the Japanese eels and the efficacy of protection against E. tarda infection was evaluated. Orally administered IgY at a dose of 400mg fish^-1 cleared E. tarda inoculated simultaneously at 10^5–6 CFU fish^-1 from the intestine within 24h. Moreover, orally administered IgY at doses of 200 and 400mg fish^-1 inhibited the penetration of E. tarda inoculated simultaneously at 10^4–6 CFU fish^-1 into the liver and kidney via the damaged intestine. The fish orally administered with IgY showed reduced mortality. These results suggest that egg yolk containing anti- E. tarda IgY is effective in preventing edwardsiellosis.     

Changes in the nutritional parameters of muscles of the common carp (Cyprinus carpio) and the silver carp (Hypophthalmichthys molitrix) following environmental exposure to cyanobacterial water bloom

The present study evaluated the effect of naturally developing cyanobacteria on the composition of muscles of two commercially important freshwater fish species. Fish were exposed to cyanobacterial biomass including Microcystis aeruginosa and Microcystis ichthyoblabe for 4 weeks. Then, they were transferred to dechlorinated potable water without any cyanobacteria for another 4-week period, thus modelling their preparation for consumers. Samples of muscles were collected every week during exposure and subsequent stay in dechlorinated potable water. The cyanobacterial water bloom of 3.9–6 × 10⁵ cells mL⁻¹ (133–383 μg g⁻¹ of total MC DW) induced statistically significant effects only in the content of fatty acids ( P <0.05; P <0.01) in the common carp ( Cyprinus carpio ), while all studied parameters including the content of dry matter and fat ( P <0.01), proteins ( P <0.05), fatty acid composition ( P <0.05; P <0.01) and some amino acids ( P <0.05) were affected in the silver carp ( Hypophthalmichthys molitrix ). This study has shown that cyanobacteria in the environment of commercially produced fish may decrease the dietetic value of fish muscles.     

Effect of methionine on intestinal enzymes activities, microflora and humoral immune of juvenile Jian carp (cyprinus carpio var. Jian)

An 8-week feeding experiment was conducted to determine the effect of dietary methionine supplementation on intestinal microflora and humoral immune of juvenile Jian carp (initial weight of 9.9 ± 0.0 g) reared in indoor flow-through and aerated aquaria. Eight amino acid test diets (350 g kg⁻¹ crude protein, CP), using fish meal, soybean-condensed protein and gelatin as intact protein sources supplemented with crystalline amino acids, were formulated to contain graded levels of methionine (0.6–22.0%) at a constant dietary cystine level of 3 g kg⁻¹. Each diet was randomly assigned to three aquaria. Growth performance and feed utilization were significantly influenced by the dietary methionine levels ( P  < 0.05). Maximum weight gain, feed intake occurred at 12 g kg⁻¹ dietary methionine ( P  < 0.05). Methionine supplementation improved hepatopancreas and intestine weight, hepatosomatic and intestine index, intestinal γ-glutamyltransferase and creatine kinase activity, Lactobacillus count, Bacillus count, lysozyme activities, lectin potency, sim-immunoglobulin M content, addiment C3,C4 contents and serum total iron-binding capacity and declined Escherichia coli and Aeromonas counts. Quadratic regression analysis of weight gain against dietary methionine levels indicated that the optimal dietary methionine requirement for maximum growth of juvenile Jian carp is 12 g kg⁻¹ of the dry diet in the presence of 3 g kg⁻¹ cystine.     

Evaluation of carbohydrate rich diets through common carp culture in manured tanks

Four diets (T₀–T₃) were formulated reducing the fishmeal (Indian) component by 100 g kg^–1 from 300 to 0 g kg^–1 and including proportionately increasing quantities of maize. Diets were fed for 120 days at 50 g kg^–1 body weight to triplicate groups of common carp (av. wt. 2.11–2.18 g) stocked at 1 m^–2 in mud bottomed cement tanks (18 m²), fertilized with poultry manure. Fish growth, SGR and FCR in the different treatments were statistically not significantly different ( P  > 0.05). PER was lowest for the 300 g fishmeal kg^–1 diet treatment (diet T₀), increasing with decrease in dietary fishmeal content (diets T₁–T₃). Fish survival ranged from 96.29 to 100%. Diets influenced carcass composition and digestive enzyme activity. A significant increase in lipid deposition was recorded with increasing dietary carbohydrate content. Amylase, protease and lipase activities were higher in fish fed with diets T₂ and T₃. The protein sparing effect of dietary carbohydrate and the economic implication of eliminating fishmeal from the diet are discussed.     

Quantitative dietary threonine requirement of juvenile Pacific white shrimp, Litopenaeus vannamei (Boone) reared in low-salinity water

An 8-week feeding trial was conducted to determine the threonine requirement of juvenile Pacific white shrimp Litopenaeus vannamei (Boone) in low-salinity water (0.50–1.50 g L⁻¹). Diets 1–6 were formulated to contain 360 g kg⁻¹ crude protein with fish meal, wheat gluten and pre-coated crystalline amino acids with six graded levels of l -threonine (9.9–19.0 g kg⁻¹ dry diet). Diet 7, which was served as a reference, contained only intact proteins (fish meal and wheat gluten). Each diet was randomly assigned to triplicate groups of 30 shrimps (0.48±0.01 g), each four times daily. Shrimps fed the reference diet had similar growth performance and feed utilization efficiency compared with shrimps fed the diets containing 13.3 g kg⁻¹ or higher threonine. Maximum specific growth rate (SGR) and protein efficiency ratio were obtained at 14.6 g kg⁻¹ dietary threonine, and increasing threonine beyond this level did not result in a better performance. Body compositions, triacyglycerol and total protein concentrations in haemolymph were significantly affected by the threonine level; however, the threonine contents in muscle, aspartate aminotransferase and alanine aminotransferase activities in haemolymph were not influenced by the dietary threonine levels. Broken-line regression analysis on SGR indicated that optimal dietary threonine requirement for L. vannamei was 13.6 g kg⁻¹ dry diet (37.8 g kg⁻¹ dietary protein).     

Effect of synthetic GnRH analogues and dopamine antagonists on the maturation of Pacific mullet, Mugil so-iuy Bas.

Abstract. The effect of two GnRH-As – des Gly¹⁰ (D-Ala)⁶ LHRH and des Gly¹⁰ (D-Tle)⁶ GnRH in total doses 10–249mu;g/kg – and four neuroleptics – pimozide, sulpiride, metoclopramide and isofloxythepin in total doses 1–50mg/kg - and environmental temperatures on the development of final stages of Pacific mullet oogenesis was investigated. The best technique of those used by us for spawning stimulation of Pacific mullet, Mugil so-iuy Bas., females is shown to be administration of neuroleptics in doses of l-8mg/kg with posterior injections on GnRH-A in doses of 25 and 50mu; g/kg applied 4 and 28h respectively after the neuroleptic injection. At water temperatures below 17°C, ovulation does not occur unless the optimal stimulation is applied.     

The fishery of the Ea Kao reservoir, southern Vietnam: a fishery based on a combination of stock and recapture, and self-recruiting populations

The present paper describes the fishery of the Ea Kao reservoir in south Vietnam. Historical data on the total production and total numbers of fish stocked were available from 1983 to 1996, and this information, together with the results of monthly monitoring of the fishery from June 1996 to December 1998, was used in the present study. A number of gears are used in the Ea Kao fishery, the most important being gill, lift and integrated nets, and beach seines. The fishery of Ea Kao is based on the annual stocking of advanced fry/early fingerlings of 0.3–1.0-g bighead carp, Hypophthalmichthys nobilis (Richardson), silver carp, Hypophthalmichthys molitrix (Cuvier & Valenciennes) and rohu, Labeo rohita Hamilton, as well as the self-recruiting indigenous species Toxabramis houdemeri Pellegrin and two exotic species, i.e. Nile tilapia, Oreochromis niloticus (L.), and common carp, Cyprinus carpio L. On average, the regularly stocked species contribute to 78% of total production, which has been around 400–450 kg ha⁻¹ in the last few years. The monthly peak catches of stocked species tend to coincide with the rainy season from May to October and the catches of self-recruiting species peak between February and April. A significant relationship ( P < 0.001) exists between the stocking density in year n (SD, n ha⁻¹) and the yield of stocked fish in year n + 1 ( Y , kg ha⁻¹):     

Insight into nucleotide and Ca<Superscript>2+</Superscript> binding to carp α-actin

ABSTRACT:   Nucleotides and Ca²⁺ binding to α-actin prepared from ordinary skeletal muscle of carp Cyprinus carpio was studied. When bound Ca²⁺ was removed with ethylenediaminetetraacetic acid, carp α-actin denatured more rapidly than chicken α-actin. Kinetic studies of the denaturation process showed that in the absence of divalent cations, the binding constants of ATP to carp and chicken actin were 5.0 × 10⁴/M and 1.2 × 10⁵/M, respectively. Competitive binding of Ca²⁺ between actin and 8-amino-2-[(2-amino-5-methylphenoxy)methyl]-6-methoxyquinoline-N,N,N',N'-tetraacetic acid (Quin 2) showed that affinity of Ca²⁺ for carp actin was also lower than that for chicken actin by a factor of 1.6. These results indicated that carp actin could relatively easily denature due to the low affinities of these ligands. Enthalpy changes upon ATP binding to carp and chicken actin were −65 kJ/mol and −110 kJ/mol, respectively. Thermodynamic analyses of our results revealed that the entropy change associated with ATP binding to carp actin was significantly smaller than that to chicken actin, suggesting that structural stabilization upon ATP binding was less effective in carp actin.     

Use of neuroactive catecholamines to chemically induce metamorphosis of hatchery-reared flat oyster, Ostrea angasi, larvae

Low numbers and unreliable wild catch of the native flat oyster, Ostrea angasi , spat has resulted in the NSW flat oyster industry being reliant on hatchery-produced spat. The need to produce culchless spat in the hatchery stimulated investigation of several catecholamines to induce metamorphosis in O. angasi larvae. Larvae were treated with one of four neuroactive catecholamines (epinephrine, epinephrine bitartrate, l -Dopa and GABA) at one of four concentrations (10⁻³, 10⁻⁴, 10⁻⁵ or 10⁻⁶  m ) for one of three treatment durations (0.5, 1–2 h) to determine morphogenic action for culchless spat production. Epinephrine bitartrate at 10⁻³ and 10⁻⁴  m and epinephrine at 10⁻³, 10⁻⁴ and 10⁻⁵  m , for a treatment duration of 1–2 h, produced significantly greater numbers of spat and culchless spat, compared with any other treatment combination. The other catecholamines tested did not induce a significant increase in the total number of spat or culchless spat, over untreated controls. Separate trials found that long-term treatment (24 h) with epinephrine bitartrate and epinephrine at morphogenic concentrations inhibited metamorphosis. Consecutive daily use of epinephrine bitartrate increased the numbers of spat and culchless spat produced, but did not affect larval or short-term post-larval survival. Treatment with 10⁻³  m epinephrine bitartrate or 10⁻⁴  m epinephrine for 1 h is recommended for routine commercial production of culchless flat oyster spat.     

Effect of dietary protein levels on growth of Penaeus stylirostris juveniles

The shrimp Penaeus stylirostris is currently produced on a commercial scale in Tahiti and New Caledonia. Both super-intensive (80 animals m⁻²) and semi intensive (25 m⁻²) systems are promoted. Locally produced commercial feed contains 380–400 g kg⁻¹ crude protein (CP) without special consideration for environmental impact. The need for a 'low pollution' diet implies reconsideration of the optimum dietary protein level for this species. Under experimental conditions, six isoenergetic practical diets ranging from 270 g kg⁻¹ to 440 g kg⁻¹ CP were formulated and fed to satiation for 30 days to juvenile P. stylirostris ; average growth rates were between 5.5 and 7.5 g per month with survival rates > 90%.
The lowest protein levels 270–310 g kg⁻¹, gave significantly ( P < 0.05) poorer growth (5.5 g per 30 days) than was observed with 330–430 g kg⁻¹ CP; 330 g kg⁻¹ CP may be recommended, and as it is lower than levels in diets used currently, there is a possibility of reducing nitrogenous waste. In addition to growth response, protein efficiency ratio, protein productive value and food conversion (feed/gain) all supported a recommended dietary protein level of 330 g kg⁻¹. Future prospects for practical feeds with even lower CP levels are considered.     

Effect of Oxygen Saturation in Water on Reproductive Performances of Pacu Piaractus brachypomus

Broodstock pacu Piaractus brachypomus as well as their eggs during their embryonic development were exposed to either normoxia (5.5–7.5 mg O₂/L) or hypoxia (2.0–4.5 mg O²/L) conditions. The plasma concentrations of 11-ketotestosterone in males and estradiol-17β in females, as well as that of their precursor testosterone (T) were significantly ( P < 0.01) higher in fish maintained under normoxic conditions than in fish exposed to hypoxia. After ovulation and spermiation induced by hormonal treatments, the plasma concentrations of T and 17,20β-dihydroxy-4-pregnen-3-one (17,20βP) significantly ( P 0.05) increased in both sexes under both normoxic and hypoxic conditions. The plasma levels of T and 17,20βP achieved under normoxic conditions were higher than the ones recorded under hypoxia, except for those of 17,20βP in males. Males responded positively to the hormonal treatments, and the concentration of spermatozoa was 10.5 ± 0.8 10⁹/mL under both oxygen conditions. Hypoxia resulted in significantly lower survival of embryos (17.3 ± 28%) in comparison to normoxic conditions (68.5 ± 25%). Moreover, larval deformities were found when exposed to hypoxia (91.6 ± 6%). During embryonic development of this species 4 mg/L of oxygen is tolerated at 26–27 C without negative impact. We conclude that despite the highly adaptable nature of adult pacu to environmental hypoxia, oxygen concentrations below 4 mg/L severely impacted survival of embryos.     

Energy and protein requirements for maintenance and growth in gilthead seabream (Sparus aurata L.)

Factorial determinations of energy and protein requirements in growing Sparus aurata were carried out at 23–24°C. The energy content in the whole fish was dependent on fish weight and ranged from 5 to 11 MJ kg⁻¹ body mass for 1–250 g fish, whereas the protein content remained constant at 179 g kg⁻¹.
During starvation the fish lost 42.5 kJ body weight (BW) (kg)^−0.83 day⁻¹ and 0.42 g protein BW (kg)^−0.70 day⁻¹. The maintenance requirement for energy was calculated to be 55.8 kJ BW (kg)^−0.83 day⁻¹ and for protein 0.86 g BW (kg)^−0.70 day⁻¹. Utilization of digestible energy and digestible crude protein below and at maintenance was determined as 0.72 and 0.51, respectively. Utilization of digestible energy and digestible crude protein for growth above maintenance was determined as 0.46 and 0.28, respectively.
These values allow estimation of requirements for growing Sparus aurata .     

Effect of different synthetic gonadotrop-releasing hormone analogues and their combinations with an anti-dopaminergic compound on the reproduction performance of sterlet (Acipenser ruthenus L.)

In this study, three synthetic gonadotrop-releasing hormones (GnRH) (azagly-nafarelin; des-Gly¹⁰-( d -Ala⁶)-LH-RH; and des-Gly¹⁰-( d -Phe⁶)-LH-RH) either alone or in combination with metoclopramide were used to induce reproduction of sterlet. The GnRH analogues were applied in a single dose of 40 μg kg⁻¹ of female and 20 μg kg⁻¹ of male body weight. Metoclopramid was administered in a simultaneous injection of 10 and 5 mg kg⁻¹ of body weight for females and males respectively. There were no significant differences in the ovulatory responses of females; ovulation rates varied between 57% and 80%, and at the temperature of 15.5–16.0 °C about 30–34 h were required for final maturation, when eggs of 17.3±1.3% of body weight were stripped. However, the fertilization rates of the des-Gly¹⁰-( d -Phe⁶)-LH-RH-treated groups were significantly lower than that in the other treatment. In males, the combination of the above peptidergic hormones with metoclopramide gave significantly better results than their single application. The results demonstrate that the final stage of gamete maturation in sterlet may be achieved by several hormonal means. The possibility of using new GnRH analogues without dopamine antagonists yields new perspectives for induced breeding of sturgeons, which have particular importance in the light of meat and roe (caviar) production for human consumption.     

Myo-inositol prevents oxidative damage, inhibits oxygen radical generation and increases antioxidant enzyme activities of juvenile Jian carp (Cyprinus carpio var. Jian)

This study was conducted to evaluate the effects of dietary myo -inositol (MI) on the antioxidant status of juvenile Jian carp ( Cyprinus carpio var. Jian). A total of 1050 Jian carp (22.28±0.07 g) were randomly distributed into seven groups of three replicates each, feeding diets containing graded levels of MI (163.5, 232.7, 384.2, 535.8, 687.3, 838.8 and 990.3 mg kg⁻¹ diet) for 60 days. Results indicated that the malondialdehyde content was the lowest for fish fed diets containing ≥384.2 mg MI kg⁻¹, and the highest for fish fed the MI-unsupplemented basal diet ( P <0.05). The protein carbonyl content was decreased with increasing dietary MI levels up to 535.8 mg kg⁻¹ diet, and no differences were found with a further increase in the MI concentration. The anti-superoxide anion capacity (ASA) and anti-hydroxyl radical capacity (AHR) were increased with increasing MI levels up to 535.8 mg kg⁻¹ diet, and plateaued thereafter. The superoxide dismutase and glutathione- S -transferase activities showed the same tendency with the ASA capacity. Catalase, glutathione peroxidase and glutathione reducase activities were improved with increasing MI levels up to 838.8, 384.2 and 687.3 mg kg⁻¹ diet, respectively, and remained nearly constant thereafter. These results suggested that MI could inhibit oxygen radical generation, increase enzymatic antioxidant capacity and prevent oxidative damage of carp. Dietary MI requirements for ASA and AHR activities of juvenile Jian carp were 567.94 and 517.22 mg MI kg⁻¹ diet respectively.     

Purification and characterization of the amylase from a small abalone <Emphasis Type="Italic">Haliotis sieboldii</Emphasis>

ABSTRACT:   Amylase, with MW of 59 kDa, was purified from small abalone Haliotis sieboldii by ammonium sulfate fractionation, CM Sepharose Fast Flow and Sephacryl S-100 HR chromatographies. The optimal pH and temperature of purified amylase were 6.0 and 37°C, respectively. The purified enzyme was stable at pH 6.0–8.0 and low temperatures. It was activated by Ba²⁺, Mg²⁺, Ca²⁺, Co²⁺, Ni²⁺, Mn²⁺, K⁺, Ag⁺, Na⁺ and Li⁺, but completely or partially inhibited by Al³⁺, Cu²⁺, Cd²⁺, Hg²⁺ and Zn²⁺. EDTA could completely inhibit, while iodoacetamide, N-ethylmaleimide and urea partially inhibit the purified amylase. According to the digestion mode of various polysaccharides, the purified enzyme was considered to be an α-amylase.     

Biochemical parameters of blood plasma and content of microcystins in tissues of common carp (Cyprinus carpio L.) from a hypertrophic pond with cyanobacterial water bloom

The aim of this study was to evaluate the dynamics of the blood plasma parameters and the content of microcystins in the tissues of common carp ( Cyprinus carpio L.) in relation to the toxic cyanobacterial water bloom. Fish (average body mass 2176±697 g) in the hypertrophic pond were exposed to natural water bloom (dominated by Planktothrix agardhii, Pseudanabaena limnetica and Limnothrix redekei ), which contained microcystins (concentration in biomass 20–181 μg g⁻¹ dry wt, concentration in water 0.3–9.5 μg L⁻¹). Biochemical parameters in fish blood plasma were analysed in 89 fish at 14-day intervals during the whole season (nine sampling periods). Our results demonstrated high variability and fluctuations in the investigated parameters. The content of microcystins and density of cyanobacterial cells correlated with some haematological indices as lipase, alanine–aminotransferase, albumin, magnesium and chlorides. The concentrations of microcystins in the muscle and liver of the fish (determined by high-performance liquid chromatography with mass spectrometer) were below the limit of detection during the monitored period [0.31 ng g⁻¹ fresh weight (f.w.) for the liver and 0.13 ng g⁻¹ f.w. for muscle]. Our study demonstrates that although known cyanobacterial toxin microcystins were not detected in the fish tissues, several biochemical parameters important for the fish physiology were modulated by natural cyanobacterial bloom.     

Perception of light intensity by Haliotis discus discus based on locomotor activity patterns

ABSTRACT:   An apparatus to measure the locomotor activity of aquatic benthic organisms at variable low light levels was developed and the diurnal behavioral pattern of the abalone Haliotis discus discus was measured at various low light intensities. During the experiment, abalone were exposed to 12 h light–dark cycles of complete darkness, 0 µmol/m²/s throughout the 12 h dark cycle and, during periods I (days 1–8) and III (days 19–26), the 12 h light cycles were set at 10 µmol/m²/s. During period II (days 10–17), abalone were exposed to a light level during the 12 h light cycles of 1 × 10⁻⁵, 1 × 10⁻⁶, 1 × 10⁻⁷ or 1 × 10⁻⁸ µmol/m²/s and the changes in locomotor activity assessed. At daytime levels of 1 × 10⁻⁵ µmol/m²/s, typical behavioral patterns were observed of high locomotory activity during the night-time cycle. However, at lower light intensities, the distinction between day and night activity patterns became less clear and, at intensities lower than 1 × 10⁻⁷ µmol/m²/s, the difference between activity during the light and dark cycles became negligible. Based on this, we conclude that the threshold of light level perception in relation to locomotor activity is approximately 1 × 10⁻⁷ µmol/m²/s. The significance of these results in relation to the entrainment of behavior in abalone is discussed.     

Identification and expression analysis of ghrelin gene in common carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis>

ABSTRACT:   A ghrelin gene has been cloned and sequenced in common carp Cyprinus carpio . Ghrelin cDNA is composed of 461 bp [with a 36-bp 5'-untranslated region (UTR) and a 113-bp 3'-UTR], which translates into a protein of 103 amino acid residues. Carp ghrelin (preproghrelin) contained a predicted signal peptide of 26 amino acid residues, the ghrelin domain ( Gly ²⁷– Val ⁴⁵) and C-terminal peptide ( Gly ⁴⁶– Phe ¹⁰³). Homology analysis of the ghrelin domain of carp with that of other known ghrelin in vertebrates showed good similarity to teleost ghrelin (50–81.8%). Hydropathy analysis based on the deduced amino acid sequence of ghrelin domains in teleosts showed a similar profile. Carp ghrelin clustered with ghrelin of goldfish Carassius auratus and other teleosts, away from mammalian, reptilian, avian, amphibian and chondrichthian ghrelin, by phylogenetic analysis. Genomic organization of carp ghrelin gene was composed of four exons and three introns, which was the same as that of other teleosts and human ghrelin genes. The carp ghrelin gene was expressed in unstimulated tissues such as foregut, hindgut, spleen and brain. In spleen cells, expression of the ghrelin gene increased upon stimulation with lipopolysaccharide (LPS), phytohemagglutinin (PHA) or imiquimod. The identification of carp ghrelin gene and the analysis of the modulation of its expression in immune-activated conditions will allow a more complete analysis of the roles of ghrelin in teleosts.