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The biomechanics of fast-starts during ontogeny in the common carp cyprinus carpio

Common carp Cyprinus carpio L. were reared a constant temperature of 20 degrees C from the larval (7 mm total length) to the juvenile (80 mm) stage. Body morphology and white muscle mass distribution were measured. Fast-start escape responses were recorded using high-speed cinematography from which the velocities, accelerations and hydrodynamic power requirements were estimated. All three measures of fast-start performance increased during development. White muscle contraction regimes were calculated from changes in body shape during the fast-starts and used to predict the muscle force and power production for all longitudinal positions along the body. Scaling arguments predicted that increases in body length would constrain the fish to bend less rapidly because the cross-sectional muscle area, and hence force production, does not increase at the same rate as the inertial mass that resists bending. As predicted, the increases in body length resulted in decreases in muscle shortening velocity, and this coincided with increases in both the force and power produced by the muscles. The hydrodynamic efficiency, which relates the mechanical power produced by the muscles to the inertial power requirements in the direction of travel, showed no significant change during ontogeny. The increasing hydrodynamic power requirements were thus met by increases in the power available from the muscles. The majority of the increases in fast-start swimming performance during ontogeny can be explained by size-dependent increases in muscle power output. For all sizes, there was a decrease in muscle-mass-specific power output and an increase in muscle stress in a posterior direction along the body due to systematic variations in fibre strain. These changing strain regimes result in the central muscle bulk producing the majority of the power requirements during the fast-start, and this power is transmitted to the tail region of the fish and ultimately to the water via muscle in the caudal myotomes.

Journal Article↗

Expression and inheritance of RSVLTR-rtGH1 complementary DNA in the transgenic common carp, Cyprinus carpio.

Transgenic common carp, Cyprinus carpio, possessing the long terminal repeat (LTR) sequence of avian Rous sarcoma virus (RSV) fused to the rainbow trout (rt) growth hormone (GH1) complementary DNA (cDNA) were produced by microinjection. Initial studies showed that the transgenic common carp transmitted the foreign DNA to a significant fraction of their progeny in three of four crosses of transgenic males with control females. These progeny grew 20 to 40% faster than their nontransgenic full siblings. In this study, additional experiments were conducted to evaluate inheritance and expression of the foreign GH gene in transgenic common carp, and the growth performance of these transgenic fish. Four P1 (parental generation produced by microinjection) x nontransgenic controls, four P1 x P1, and one P1 x F1 matings of transgenic carp containing RSVLTR-rtGH1 cDNA were made. The percentages of transgenic progeny resulting from these matings were: 0, 32, 42, 100 (4 progeny only), 21, 21, 31, 30, and 23%, respectively. All crosses except 1 siblot (control x P1) exhibited progeny ratios below the expected 50 or 75% transgenic. These results indicate that most of these transgenic P1 had the foreign gene in their germ line but were mosaics, and at least one transgenic individual did not have the RSVLTR-rtGH1 cDNA in the gonadal tissue. Both P1 and F1 transgenic fish produce trout growth hormone mRNA and polypeptide as determined by reverse transcription polymerase chain reaction amplification, RNA dot-blot hybridization, and radio-immunobinding assay. Growth response by families of F1 transgenic fish to the addition of rtGH1 cDNA varied widely.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Plasticity of myosin heavy chain expression with temperature acclimation is gradually acquired during ontogeny in the common carp (Cyprinus carpio L.).

Common carp (Cyprinus carpio L.) were reared from hatching until 61 mm total length (TL) at 21 degrees C. At 14 weeks and 20 weeks post-hatch, corresponding to initial lengths of 30 mm and 44 mm respectively, fish were acclimated to 10 degrees C using a rate of cooling of 1 degrees C per day. A statistical model was used to compare the time course in the change of white muscle myofibrillar ATPase activity with temperature acclimation. The myosin heavy chain (MHC) composition of white muscle myofibrils was investigated using peptide mapping. A significant increase in myofibrillar ATPase activity was observed after 2-3 weeks in the 44 mm group, but not until 4-5 weeks in the 30 mm group. when they had reached 37 mm TL. The MHC banding pattern of 120 mm TL fish acclimated to 10 degrees C or 21 degrees C for a minimum of 6 weeks were distinct from each other. The MHC peptide map characteristic of 10-degrees C-acclimated fish was not observed in individuals less than 37 mm length. We therefore conclude that the capacity to alter the composition and properties of myofibrils with cold acclimation is acquired in juvenile carp at around 37 mm TL.

Acclimatization↗

Temperature and the expression of myogenic regulatory factors (MRFs) and myosin heavy chain isoforms during embryogenesis in the common carp Cyprinus carpio L.

Embryos of the common carp, Cyprinus carpio L., were reared from fertilization of the eggs to inflation of the swim bladder in the larval stage at 18 and 25 degrees C. cRNA probes were used to detect transcripts of the myogenic regulatory factors MyoD, Myf-5 and myogenin, and five myosin heavy chain (MyHC) isoforms during development. The genes encoding Myf-5 and MyoD were switched on first in the unsegmented mesoderm, followed by myogenin as the somites developed. Myf-5 and MyoD transcripts were initially limited to the adaxial cells, but Myf-5 expression spread laterally into the presomitic mesoderm before somite formation. Two distinct bands of staining could be seen corresponding to the cellular fields of the forming somites, but as each furrow delineated, Myf-5 mRNA levels declined. Upon somite formation, MyoD expression spread laterally to encompass the full somite width. Expression of the myogenin gene was also switched on during somite formation, and expression of both transcripts persisted until the somites became chevron-shaped. Expression of MyoD was then downregulated shortly before myogenin. The expression patterns of the carp myogenic regulatory factor (MRF) genes most-closely resembled that seen in the zebrafish rather than the rainbow trout (where expression of MyoD remains restricted to the adaxial domain of the somite for a prolonged period) or the herring (where expression of MyoD persists longer than that of myogenin). Expression of two embryonic forms of MyHC began simultaneously at the 25-30 somite stage and continued until approximately two weeks post-hatch. However, the three adult isoforms of fast muscle MyHC were not detected in any stage examined, emphasizing a developmental gap that must be filled by other, as yet uncharacterised, MyHC isoform(s). No differences in the timing of expression of any mRNA transcripts were seen between temperature groups. A phylogenetic analysis of the MRFs was conducted using all available full-length amino acid sequences. A neighbour-joining tree indicated that all four members evolved from a common ancestral gene, which first duplicated into two lineages, each of which underwent a further duplication to produce Myf-5 and MyoD, and myogenin and MRF4. Parologous copies of MyoD from trout and Xenopus clustered closely together within clades, indicating recent duplications. By contrast, MyoD paralogues from gilthead seabream were more divergent, indicating a more-ancient duplication.

Animals↗

A study of sequential histopathology of Trypanoplasma borreli (Protozoa: Kinetoplastida) in susceptible common carp Cyprinus carpio.

The tissue response of common carp Cyprinus carpio to the kinetoplastid blood parasite Trypanoplasma borreli Laveran & Mesnil, 1901 was investigated during a laboratory infection of a highly susceptible carp line. With the development of the parasitaemia an increased proliferation of the lymphoid renal interstitial tissue was induced, which resulted in a progressive depression and deterioration of renal tubules. In heavily infected carp at Days 20 to 28 post inoculation (PI), a tubulonephrosis, a glomerulitis caused by a massive accumulation of leukocytes in glomerular capillaries, and large numbers of trypanoplasms in blood vessels and renal interstitium were observed. Corresponding with rising T. borreli numbers in the peripheral blood, splenic lymphocytes showed increasing proliferation rates, and the capillaries of the liver, gills, heart and intestine were infiltrated with lymphocytes and trypanoplasms. In heavily infected carp, congestion of liver sinusoids, focal necroses of hepatic tissue, extensive accumulations of erythrocytes in the spleen and in the blood marked anaemia were observed. These carp often showed abdominal distension, exophthalmus and swimming disorders described as 'sleeping sickness of carp'. Proliferation of cells from the interstitial lymphoid tissue of the kidney, which bears a close resemblance to the bone marrow of higher vertebrates, is considered a normal immune response of fish to antigen challenge. We here describe the unique case of a severe but ineffective immune reaction which results in the destruction of excretory renal structures. This has to be considered a severe disturbance of osmoregulation in affected carp, which, together with a decrease in oxygen uptake due to anaemia, is likely a major cause of death in these carp.

Animals↗

Genetic control of glucosephosphate isomerase (GPI) in common carp (Cyprinus carpio L.).

In common carp, a freshwater fish species of tetraploid origin, GPI enzymes are present in two variants: GPI-A and GPI-B. GPI-A is coded by two loci segregating for two (GPI-A 1*) and six (GPI-A2*) alleles. Experimental crosses of the ornamental (Koi) variety of common carp revealed that GPI-B is coded by only one locus (GPI-B*). Another GPI-B* locus must have been silenced in the process of functional diploidization. It was also shown that the GPI-A2* locus segregated independently from the GPI-B* locus, demonstrating that the loci are located on different chromosomes.

Alleles↗

An unusual koi herpesvirus associated with a mortality event of common carp Cyprinus carpio in New York State, USA.

Koi herpesvirus (KHV), a highly contagious and lethal virus that affects both koi (Cyprinus carpio koi) and common carp (Cyprinus carpio), was isolated in 1998 from two outbreaks of koi suffering mass mortality in New York State, USA, and in Israel. The disease had been described as early as 1996 in Europe. In July 2004, this virus was found associated with a mass mortality event in wild common carp in the Chadakoin River, New York, USA (42 degrees 07' N, 79 degrees W). Affected fish typically showed marked hyperplasia of gill tissues, abdominal adhesions, and severe multifocal to diffuse external hemorrhages. The virus isolated in this outbreak was somewhat unusual in that it initially replicated well in fathead minnow cell cultures, which is typical of spring viremia of carp virus. Testing at the National Veterinary Services Laboratories, Ames, Iowa, USA, confirmed the virus's identity to be KHV. Koi herpesvirus is not currently on the OIE (World Organisation for Animal Health) list of notifiable diseases; however, it is capable of causing mass mortality in susceptible fish at permissive temperatures.

Animals↗

Effect of certain toxicants on gonadotropin-induced ovarian non-esterified cholesterol depletion and steroidogenic enzyme stimulation of the common carp Cyprinus carpio in vitro.

Isolated ovarian tissues from the common carp, Cyprinus carpio were incubated in vitro to obtain a discrete effect of four common toxicants of industrial origin, namely phenol, sulfide, mercuric chloride and cadmium chloride, on gonadotropin-induced alteration of nonesterified and esterified cholesterol and steroidogenic enzymes, delta 5-3 beta-HSD and 17 beta-HSD activity. Stage II ovarian tissue containing 30-40% mature oocytes were shown to be most responsive to gonadotropins in depleting only nonesterified cholesterol moiety and stimulating the activity of both. Safe doses of above mentioned toxicants when added separately to stage II ovarian tissue with oLH (1 microgram/incubation) gonadotropin-induced depletion of nonesterified cholesterol and gonadotropin-induced stimulation of the activity of both enzymes was significantly inhibited. Esterified cholesterol remained almost unaltered. Findings clearly indicate the impairment of gonadotropin induced fish ovarian steroidogenesis by the four toxicants separately.

17-Hydroxysteroid Dehydrogenases↗

Comparative biotransformation of molinate (Ordram) in the white sturgeon (Acipenser transmontanus) and common carp (Cyprinus carpio).

1. Juvenile white sturgeon (Acipenser transmontanus) and common carp (Cyprinus carpio) were exposed to 100 micrograms/l [ring-14C]molinate in a flow-through metabolism system. Exposure was in three phases: acclimation (2 h), uptake (24 h), and depuration (24 h). 2. Excreted metabolites were collected on a macroreticular resin, and retained metabolites were extracted from homogenized fish tissue. Identification and quantification was by h.p.l.c. cochromatography (gradient conditions) and determination of 14C, and confirmation was by t.l.c. 3. 14C depuration (elimination) by common carp (77.8%) was significantly slower than that by white sturgeon (96.0%, P less than 0.01) or that previously reported for striped bass (90.5%, P less than 0.01). Differences in bioconcentration were not significant (P less than 0.05). 4. Common carp and white sturgeon oxidized molinate to form several products and hydrolysed, or conjugated with glutathione (GSH), the sulphoxide or sulphone; both fish also formed a D-glucuronic acid conjugate. 5. Common carp were significantly less capable of molinate sulphoxidation and GSH conjugation than either white sturgeon (P less than 0.01) or striped bass (P less than 0.05). 6. The higher toxicity of molinate in common carp may be due to greater bioconcentration, slower depuration, and less efficient metabolic deactivation.

Animals↗

Responses of molecular indicators of exposure in mesocosms: common carp (Cyprinus carpio) exposed to the herbicides alachlor and atrazine.

Common carp (Cyprinus carpio) were treated in aquatic mesocosms with a single pulse of the herbicides atrazine or alachlor to study the bioavailability and biological activity of these herbicides using molecular indicators: Liver vitellogenin gene expression in male fish for estrogenic activity, liver cytochrome P4501A1 gene expression, and DNA damage in blood cells using the single-cell gel electrophoresis method. Both alachlor and atrazine showed dose-related increases in DNA strand breaks at environmentally relevant concentrations (<100 ppb). Gene expression indicators showed that neither herbicide had estrogenic activity in the carp, whereas atrazine at concentrations as low as 7 ppb induced cytochrome P4501A1. These results support the study of molecular indicators for exposure in surrogate ecosystems to gauge relevant environmental changes following herbicide treatments.

Acetamides↗

Molecular cloning and expression of preproadrenomedullin gene from common carp Cyprinus carpio L.

A preproadrenomedullin (preproAM) gene was isolated from common carp Cyprinus carpio L. by PCR mediated homology cloning. The isolated gene was composed of 516bp, which translated to a protein having 171 amino acid residues. The signal peptide was composed of 23 amino acid residues and predicted adrenomedullin (AM) was conserved in this sequence and made up of 49 amino acid residues. AM gene from carp shared identities of 64.0, 44.2, and 46.0% to takifugu (AM-1), human, and rat AM orthologues, respectively. The analysis of genomic structure revealed that carp preproAM gene is spread over two exons interrupted by one intron (92bp). AM genes clustered together and carp AM gene showed similarity to takifugu AM-1 gene according to the phylogenetic analysis. Expression analysis revealed that carp AM gene expressed constitutively in all the organs confirmed. Moreover, we demonstrated that expression of carp AM gene increased by treatment of LPS or hydrocortisone in different tissues by quantitative real-time PCR.

Amino Acid Sequence↗

Immunomodulatory effects of alpha melanocyte stimulating hormone on common carp (Cyprinus carpio L.).

The immunomodulatory effects of phagocytic cells in common carp Cyprinus carpio L., by alpha melanocyte stimulating hormone (alpha-MSH) was analyzed in vitro. Carp head kidney leucocytes were cultured in RPMI 1640 medium containing 0.05, 0.1, 1 and 10 nM alpha-MSH and the production of superoxide anion was measured via the reduction of nitroblue tetrazolium (NBT) in vitro. Macrophages incubated with alpha-MSH showed an increase in the production of superoxide anion in comparison to control macrophages cultured without hormone. Phagocytic cells treated with alpha-MSH also displayed increased phagocytosis. Furthermore, carp lymphocytes treated with alpha-MSH increased the mitogenic responses to phytohaemoagglutinin (PHA-P). These results show that alpha-MSH directly influences fish immune responses.

Animals↗

Cloning, characterization and expression analysis of interleukin-10 from the common carp, Cyprinus carpio L.

Interleukin (IL)-10 was cloned from the common carp (Cyprinus carpio L.) using IL-10 primers from carp head kidney following stimulation with concanavalin A and lipopolysaccharide. The cDNA consisted of a 1096 bp sequence containing a 55 bp 5' untranslated region and a 498 bp 3' untranslated region. An open reading frame of 543 bp encoded a putative 180 amino acid protein with a putative signal peptide of 22 amino acids. The signature motif of IL-10 is conserved in carp sequence. A 2083 bp genomic sequence of carp IL-10 was found to contain five exons interrupted by four introns. With the exception of much more compact introns, the genomic structure was similar to that of mammalian IL-10. By homology, phylogeny and genomic analyses, the carp gene cloned was designated as IL-10. Carp IL-10 was expressed in head, kidney, liver, spleen and intestine during the resting phase. The gene was also expressed in head kidney and liver following in vitro stimulation with lipopolysaccharide.

Amino Acid Sequence↗

The biomechanics and evolutionary significance of thermal acclimation in the common carp Cyprinus carpio.

The effects of thermal acclimation were investigated in the common carp Cyprinus carpio L. Acclimation and acute temperature effects were tested during ontogeny from larval [9.5 mm total length (L)] to juvenile (69.0 mm L) stages and between 8 and 21 degrees C. The myosin heavy chain (MHC) composition, myofibrillar Mg(2+)-Ca(2+)-ATPase activity, and muscle strains showed significant thermal acclimation effects. MHCs were only expressed in an acclimation temperature-dependent fashion in fish longer than 37 mm. During fast starts, the temperature had a significant effect on the white muscle strain (33% increase and 50% decrease with increasing acclimation and acute temperature, respectively) and contraction duration (25% decrease with increasing acute temperature). Increases in hydrodynamic efficiency (0.19 to 0.38) and hydrodynamic power requirements (Q(10) = 3.2) occurred with increasing acute temperature (10 to 20 degrees C). Competing hypotheses about the evolutionary significance of the temperature acclimation response were tested. Acclimation extended the temperature range for fast-start behavior, but no improvements in performance at the whole animal level were found between 8 and 21 degrees C.

Acclimatization↗

[Genetic diversity and differentiation of Russian common carp (Cyprinus carpio L.) breeds inferred from RAPD markers].

Polymorphic components of the common carp Cyprinus carpio L. genome were examined by means of polymerase chain reaction with random primers (RAPD-PCR). Using four primers, genetic diversity estimates were obtained for 12 populations and seven strains of Russian common carp breeds, as well as for European Hungarian common carp and Amur wild common carp (N = 87). The highest number of polymorphic loci was revealed in Angelinskii common carp, as well as in the samples of Altai common carp and Amur wild common carp (P = 23.8-18.7%), while the lowest number of polymorphic loci was in the BB strain of Ropsha common carp. The index of genetic diversity, H, was high (11%) in Amur wild common carp, as well as in Altai and Angelinskii common carps. In the remaining breeds, the value of this index varied from 4 to 8%. Based on summarized RAPD profile (132 bands), a dendrogram of genetic differences was constructed. In this dendrogram, all breeds examined grouped into two clusters. One of the clusters was formed by Hungarian and Angelinskii common carps, and the three samples of Altai common carp. The second cluster was formed by the group consisting of the representatives of Cherepetskskii, Stavropol, and Ropsha common carps, along with the differing from them Amur wild common carp. The observed differentiation was confirmed by the analysis of the polymorphic markers variance by the method of principle components. Evolutionary history and the reasons for genetic differentiation of Russian common carp breeds are discussed.

Animals↗

The complete nucleotide sequence of the growth-hormone gene from the common carp (Cyprinus carpio).

We have isolated and sequenced a phase clone from a common carp (Cyprinus carpio) genomic library that carries a gene encoding growth hormone (GH). This gene consists of five exons and four introns spanning a region of about 3 kilobase pairs. Its exons correspond with one of two reported cDNAs of carp GH except for nine differences in the nucleotide sequence, while the encoded amino-acid sequences are identical. The sequence upstream from the transcription start point contains two tandem repeats of AACTCTCATG (from -85 to -62) and the typical TATA box. All the introns start with a consensus GT dinucleotide and end with AG. The arrangement of exons and introns is very similar to that seen in mammalian GH, but quite different from the GH genes of rainbow trout and Atlantic salmon.

Amino Acid Sequence↗

Response of hexokinase enzymes and the insulin system to dietary carbohydrates in the common carp, Cyprinus carpio.

The response of the common carp to diets with varying amounts of digestible starch, provided either as pea meal (LP, HP, 30 and 46% peas, respectively) or as cereal (LW, HW, 30 and 46% wheat, respectively), was studied and compared with the response to a carbohydrate-free protein-rich diet (CF). Here we focused on the utilisation of dietary carbohydrates by examining the relationship between dietary starch intake, hepatic hexokinase activities, circulating insulin and muscle insulin receptor system. Plasma glucose concentration and hepatic high Km hexokinase (glucokinase, GK) activity were not affected by the content of digestible starch, but 6 h after feeding enzyme activity was higher in the fish fed carbohydrate diets. Similarly, low Km hexokinase (HK) activity was also higher in the fish 24 h after feeding. Fat gain and protein retention were significantly improved by increased digestible starch intake, especially in the HP group, which in turn, presented the highest plasma insulin levels. Glycogen stores were moderately increased by the ingestion of digestible starch. The number of insulin receptors was greater in the CF group than in fish on carbohydrates, except the HP group. Our results confirmed that the common carp uses dietary carbohydrates efficiently, especially when there are provided by peas. This efficiency might be related to the enhanced response of postprandial insulin observed in the HP group.

Animals↗

Toxic effects of chromic sulphate on the common carp, Cyprinus carpio.

The effects of chromic sulphate, which is commonly used in the tanning of leather, on the common carp, Cyprinus carpio and the bighead, Aristichthys nobilis, were studied. C. carpio was found to be more susceptible than A. nobilis to chromic sulphate at the lowest concentration tested (100 mg/l) whereas results obtained at other concentration were similar for both species. Damage to the gills, liver and intestine was observed by histopathological examination. A wide range of chemicals is used in the different stages of treating leather and a possible treatment of the effluent from the tanneries is suggested.

Animals↗