Functional changes of the LH-immunoreactive cells in adenohypophysis of the common carp (Cyprinus carpio) from rivers contaminated with estrogenic chemicals

The adverse effect of estrogenic chemicals on luteinizing hormone-immunoreactive (LH-ir) cells in the adenohypophysis of common carp (Cyprinus carpio) was examined using immunocytochemical and morphometric methods. Adult male fish were collected from two contaminated sites (Ishizu and Wada Rivers) and from a control pond at Agriculture, Food and Environmental Sciences, Research Center of Osaka Prefecture. The concentration of nonylphenol, bisphenol A and 17beta-estradiol in the Ishizu River was 3-4 times higher than that in the Wada River. The proportion and size of LH-ir cells were evaluated using the point-counting method by optical microscopy. In control carp, the proportion of LH-ir cells in the breeding season was significantly lower than in the pre- and post-breeding seasons. The same tendency was also found in Ishizu and Wada River carp, but without statistical significance. The proportion of LH-ir cells in Ishizu River carp was significantly lower than that of the control and the Wada River in all seasons. The LH-ir cells in control carp increased in size in the breeding season. LH-ir cells in Ishizu River carp were significantly (p<0.05) smaller than those in control fish, but not different from Wada River carp. A disturbance in the secretory function of LH-ir cells was found in carp from the Ishizu River; granulation and vacuolation were not in synchronization with those of control and Wada River fish. Our data suggest that the estrogenic chemicals in the Ishizu River interfere with functions of LH-ir cells directly or through the testis.     

Myxobolus infections of common carp (Cyprinus carpio) in Syrian fish farms

During a survey on Myxobolus infection of pond-cultured common carp in Syria three Myxobolus spp. were found. Myxobolus dispar infected the gill arteries, forming large elongated plasmodia in the gill filaments. The plasmodia of M basilamellaris were located in the gill arches at the base of the filaments. Elongated filiform plasmodia of M. encephalicus were found in the blood vessels of the brain. Despite the common occurrence of the above parasites, no disease symptoms were observed in the infected fish specimens. This is the first report on myxosporean infection of fish from Syrian waters.     

Fumonisin B1 causes multiple lesions in common carp (Cyprinus carpio)

Year-1 carp were fed ratios containing 100mg/kg and 10 mg/kg of added fumonisin B1 for 42 days. The experimental and control fish were examined clinically during the experiment and at the end all fish were necropsied and histological changes recorded. Blood vessels, liver, exocrine and endocrine pancreas, excretory and haematopoietic kidney, heart and brain were sensitive both to 100 and 10mg/kg of FB1 in the diet and the rodlet cell (RC) frequency was considerably increased in and around damaged tissues. Many damaged blood vessels contained stacks of RCs above the endothelium. Other changes subsequent to fumonisin exposure that have not been previously reported include scattered lesions in the exocrine and endocrine pancreas, and interrenal tissue, probably due to ischemia and/or increased endothelial permeability. Presented findings indicate the need for more intensive studies of fumonisin-induced toxicity in cultured fish.     

Differences between the European carp (Cyprinus carpio carpio) and the coloured carp (Cyprinus carpio haematopterus) in susceptibility to Thelohanellus nikolskii (Myxosporea) infection

Thelohanellus nikolskii infection of the common carp (Cyprinus carpio L.) has been a common parasitosis in the Central European fish farms since the first detection of the parasite about 20 years ago. This parasite, introduced from the Far East, causes intensive infection on the fins of fingerlings of the carp subspecies cultured in Europe (European carp, Cyprinus carpio carpio). This infection of the common carp occurs in the Hungarian fish farms every year. Until the present study, this parasite had not been recorded from the fins of koi or coloured carp (Cyprinus carpio haematopterus), a carp of Far Eastern origin, which is cultured in Hungary as an ornamental fish. A recent survey conducted in common carp, koi and goldfish stocks demonstrated that T. nikolskii infection of low prevalence and intensity occurs also in koi populations, but its prevalence and intensity are markedly lower than in common carp kept in the same ponds. It is suggested that the observed differences are due to disparities in the susceptibility of the two carp subspecies to T. nikolskii, and that the koi is less susceptible to this infection. Other signs of susceptibility can also be observed in the European subspecies, since in 15% of the fish plasmodium development was arrested at an early stage. Thelohanellus nikolskii infection could not be demonstrated on goldfish (Carassius auratus).     

Pox-like lesions in a colony of carp (Cyprinus carpio)

Twelve carp (Cyprinus carpio) out of a total of 100 were affected with skin nodules, mainly located on the dorsal and lateral surface. Post mortem examination of six of the worst affected fish showed the nodules consisting of white gel-like outgrowths. No abnormality was apparent in any other tissue examined. Microscopically, the nodules were formed by proliferating hyperplastic epidermal cells. The other six fish were left for recovery and their nodules regressed spontaneously. The possible aetiological factors are listed.     

Herpesvirus-associated papillomas in koi carp (Cyprinus carpio).

From January through November 1994, 32% (7/22) of koi carp (Cyprinus carpio) maintained in indoor aquariums developed proliferative cutaneous lesions that consisted of single to multiple 2-10-mm whitish to pink fleshy masses usually associated with fin rays. Although scaleless koi were more commonly affected (3/6) than were normally scaled koi (4/16), the difference in incidence rates was not significant (chi2 text, P > 0.05). Lesions typically resolved spontaneously in 1-3 wk, occasionally persisted for >3 mo, and recurred in several fish after 2-5 mo. Fish were otherwise asymptomatic. Wet mount preparations from lesions were densely cellular and consisted of hyperplastic epidermal cells of normal morphology without parasites or inflammatory cells. Histologically, biopsies were consistent with papillomas and were characterized by a marked benign epidermal hyperplasia without inclusion bodies or inflammatory infiltrate. Transmission electron microscopic examination revealed intranuclear and intracytoplasmic herpesvirus virions. Virus isolation attempts were unsuccessful.     

Pharmacokinetics and pharmacodynamic effects in koi carp (Cyprinus carpio) following immersion in propofol

Objective

To test the hypothesis that plasma propofol concentration (PPC) is associated with anesthetic effect in koi carp administered propofol by immersion.

Morphology of blood cells in carp (Cyprinus carpio L.)

The morphology of blood cells in the carp was investigated by light and electron microscopy. Erythrocytes, thrombocytes, lymphocytes, granulocytes and monocytes were identified as the peripheral blood cells. Thrombocytes were round to long oval, each containing vesicular and microtubular structures and an oval nucleus with abundant heterochromatins. Lymphocytes were divided into three types in size, small, medium and large. Some of the small and medium lymphocytes were alpha-naphthyl-acetate esterase (ANAE) positive, while large lymphocytes were pyroninophilic. Granulocytes were distinguished into three types (type I, type II and type III) according to the morphology of the nucleus and granules. Type I granulocytes possessed lobulated nuclei and a large number of cytoplasmic granules, each of which was oval and contained electron-dense materials and a crystalloid. Type II granulocytes had small eccentric nuclei and were subdivided into IIa and IIb granulocytes by electron microscopic analysis. Granules of type IIa granulocytes were furnished with an electron-dense rim. Granules of type IIb granulocytes were larger than those of type IIa, containing randomly distributed electron-dense and electron-lucent materials. Type III granulocytes possessed round nuclei and a few large granules. The granules were filled with regularly arranged fibriform materials and some needle-like structures. Monocytes were morphologically similar to those of mammals.     

Surgical removal of an ovarian tumour in a koi carp (Cyprinus carpio)

Ultrasonography, radiography and exploratory coeliotomy were used to diagnose and treat a large intracoelomic neoplasm from a female koi carp (Cyprinus carpio) presented for abdominal enlargement of several months duration. Feed was withheld for 1 week immediately prior to surgery and the fish was sedated with isoeugenol (AQUI-S) at a dose rate of 10 mL/L to facilitate diagnostic imaging techniques. Surgical anaesthesia was induced by adding tricaine (MS-222) 50 mg/L to the water and an exploratory coeliotomy and tumour removal was performed. The fish was allowed to recover in fresh water at 18 degrees C and salt was added slowly to the water over a period of 1 hour to a concentration of 5 g/L This concentration was maintained in a recovery pond for 1 week postoperatively. Enrofloxacin was administered intramuscularly (10 mg/kg) immediately, 3 days and 1 week postoperatively. A diagnosis of undifferentiated ovarian carcinoma was made on the basis of the histological appearance of the neoplasm and immunohistochemical staining.     

Evaluations of the nutritional value of Jatropha curcas protein isolate in common carp (Cyprinus carpio L.)

Jatropha curcas seeds are rich in oil and protein. The oil is used for biodiesel production. Jatropha seed cake (JSC) obtained after oil extraction is rich in protein; however, it is toxic (phorbol esters content 1.3 mg/g) and consists of 50–60% shells, which are indigestible. The principle of isoelectric precipitation was used to obtain Jatropha protein isolate (JPI) from JSC and it was detoxified (DJPI). Carp (n = 45, 20.3 ± 0.13 g) were randomly distributed into five groups with three replicates and for 12‐week fed iso‐nitrogenous diets (crude protein 38%): Control [fishmeal (FM)‐based protein]; J₅₀ and J₇₅ (50% and 75% of FM protein replaced by DJPI); S₅₀ and S₇₅ (50% and 75% of FM protein replaced by soy protein isolate). Growth performance and nutrient utilisation parameters were highest in S₇₅ group and not significantly different to those in J₅₀ and S₅₀ groups but were significantly higher than those for all other groups. Similar trend was observed for protein and energy digestibilities of experimental diets, whereas opposite trend was observed for the feed to gain ratio. Activities of intestinal digestive enzymes did not different significantly between the five groups. In conclusion, DJPI is a good quality protein source for carp.     

Blood changes in carp (Cyprinus carpio) induced by ulcerative Aeromonas salmonicida infections

Only recently Aeromonas salmonicida has been recognized as a significant bacterial pathogen in ulcerative disease of cyprinid fish. Our attempts to formulate a vaccine based on bacterial surface antigens were unsuccessful in conferring reliable protection against lethal challenge. This lead us to study pathological changes in the humoral defense system during ulcerative A. salmonicida infection in carp. High numbers of opportunist pathogens such as A. hydrophila and Pseudomonas sp. were frequently recovered from the internal organs of moribund fish, in addition to A. salmonicida. These findings together with leucopenia in moribund fish suggest that pathogenesis is characterized by a state of immune suppression. In addition, fish which had sustained a sublethal infection were not protected against a subsequent lethal challenge. However, fish previously injected with a concentrated and inactivated culture supernatant showed protection. Differential blood cell counts did not differ between experimental and control groups during sublethal infection in contrast to serum proteins. Furthermore infected non-immune carp showed a progressive decrease of immunoglobulin and total serum protein levels before the day of peak mortality whereas protected carp maintained the immunoglobulin concentration despite a decrease in protein. Our observations suggest the involvement of multiple pathogenic events, affecting different parts of the humoral defense system during ulcerative A. salmonicida infection. The immunosuppressive effects can be minimized by prior vaccination with culture supernatant.     

Starvation-induced pathobiology in the gut of carp (Cyprinus carpio L.)

In order to assess the effects of starvation on intestinal intraepithelial lymphocytes (i-IEL) count and histological changes of the carp gut mucosa, one group of fish (n = 10) were fed commercially prepared standard diets and another group of fish (n = 10) were starved for 4 weeks. Carp starved for 4 weeks developed enteropathy, comprising folds atrophy, stratum compactum hyperplasia, significant periodic-acid-Schiff (PAS)-positive (P < 0.00001), but not Alcian blue (ALB)-positive, goblet cell (GC) hyperplasia and a significant decrease (P < 0.00001) in i-IEL numbers. These changes were associated with a dense cellular infiltrate into the lamina propria. Taken together, these data suggest that the pathobiology of starvation-induced i-IELs decrease, matching PAS-positive goblet cell proliferation and inflammatory cells homing to the gut, could be classified as a non-infectious enteropathy induced by starvation.     

Some observations on the narcotizing ability of electric currents on the common carp Cyprinus carpio

Some effects of alternating current electronarcosis and of rectified current electronarcosis on C. carpio were investigated. In all instances recovery from narcosis was accompanied by convulsive spasms. Haemorrhaging of the gills was also observed to occur. Carp do not appear to be suitable candidates for electronarcosis.     

Molecular phylogeny of the kidney-parasitic Sphaerospora renicola from common carp (Cyprinus carpio) and Sphaerospora sp. from goldfish (Carassius auratus auratus)

The genetic relatedness of two kidney-parasitic Sphaerospora species was studied. Although S. renicola, the causative agent of swimbladder inflammation of common carp fingerlings (Cyprinus carpio), and Sphaerospora sp. originating from goldfish (Carassius auratus auratus) were indistinguishable on the basis of spore morphology, they were found to be genetically different as their 18S rDNA sequences shared only 71.9% identical nucleotides. In the phylogenetic trees, Sphaerospora sp. from goldfish grouped with Myxidium truttae (AJ582061) within the clade of the coelozoic freshwater species. Sphaerospora renicola clustered with S. molnari (AF378345) within the group of myxosporeans histozoic in gills. The topology of the six Sphaerospora species on the phylogenetic trees implied that myxospore morphology does not correlate with the genetic relationships, and the genus seems to be polyphyletic.     

Pre-treatment of dietary plant feedstuffs with phytase and its effect on growth and mineral concentration in common carp (Cyprinus carpio L.)

Effect of phytase pre-treatment of dietary plant feed stuffs (PF) on growth and mineral concentration in common carp (C. carpio L.) was investigated. Diets used were: C0, diet with no supplement; CI, diet with incubated PF; CP0, diet supplemented with 3 g P/kg; CPI, diet with incubated PF supplemented with 3 g P/kg; Phyt0, diet fortified with 4000 U phytase/kg; PhytI, diet with PF incubated with 4000 U phytase/kg. Mean weight gain, SGR and FCR were the same (p < 0.05) in fish fed CP0, CPI or PhytI but less (p < 0.05) in other fish. Bone P was similar in fish fed CP0 (74.9), CPI (75.9) or PhytI (71.5 mg/g DM) but higher (p < 0.05) than in fish fed C0. Bone Ca and Mg were similar in fish fed CP0, CPI or PhytI but less (p < 0.05) in other fish. Bone Zn in fish fed C0, Phyt0 or PhytI was higher (p < 0.05) than that in fish fed CP0 or CPI. In conclusion, CP0, CPI and PhytI had the same effect, suggesting phytase pre-treatment as the most effective method. CPI did not have any advantage over CP0. Phytase increased mineral availability and utilization, which would minimize cost of mineral supplementation and discharges into the environment.     

Effects of solid-state fermentation product of yeast supplementation on liver and intestinal health,and resistance of common carp(Cyprinus carpio)against spring viraemia carp virus

This study aimed to investigate the effects of solid-state fermentation products of yeast(SFPY)on liver and intestinal health and disease resistance of common carp(Cyprinus carpio).A total of 200 common carp with an initial average weight of 2.55±0.004 g were divided into 5 groups(4 replications per group and 10 fish per replication),and were fed with one of five diets,including a control diet and 4 diets supplemented with 2‰(Y2),3‰(Y3),4‰(Y4),or 5‰(Y5)SFPY,respectively,for 8 weeks.Results indicated that,the addition of SFPY to the diet of common carp did not affect the growth performance or survival rate of fish(P=0.253).Interestingly,with the addition of SFPY,the triacylglycerol(TAG)content of the liver presented a linear decreasing tendency(P=0.004),with significantly decreased in Y4 and Y5 groups(P=0.035)compared with control.Serum lipopolysaccharide(LPS)content and diamine oxidase(DAO)activity presented a negative linear relationship with the addition of SFPY(P=0.015,P=0.030),while serum lipopolysaccharide binding protein(LBP)content first decreased and then increased(P＜0.001).The total antioxidant capacity(T-AOC)in the intestine of fish increased continuously with increasing SFPY supplementation(P=0.026),reaching the highest level in Y5 group.The villus height in all experimental groups were significantly higher than that in the control group(P＜0.001).Furthermore,compared to the control,adding 3‰ SFPY to the control diet of common carp significantly increased the relative abundance of Fusobacteria(P=0.018)and decreased that of Proteobacteria(P=0.039)at phylum level,and increased the relative abundance of Cetobacterium(P=0.018)and decreased that of Shewanella(P=0.013)at genus level.Compared with the control,the relative mRNA expression level of spring viraemia of carp virus N protein(SVCV-n)in the kidney was lower than that of the control group without significance and bottomed out in Y4 group(P=0.138).In conclusion,dietary SFPY enhanced the SVCV resistance capacity of common carp by improving liver and intestinal health and modulating the gut microbiota.Thus,SFPY is a potential feed additive to be used in aquaculture to reduce the huge economic loss of common carp due to SVCV disease.Based on liver TAG content and intestinal villus height,the optimal addition level of SFPY was 3.02‰ and 2.72‰,respectively.     

Technical note: viability and motility of vitrified/thawed primordial germ cell isolated from common carp (Cyprinus carpio) somite embryos

The feasibility of cryopreserving common carp (Cyprinus carpio) primordial germ cells (PGC) by vitrification of whole embryos at the 22- to 28-somite stage was investigated. Green fluorescent protein (GFP)-labeled PGC were cooled rapidly using liquid nitrogen after exposure to a pretreatment solution containing 1.5 M cryoprotectant (ethylene glycol or dimethyl sulfoxide, 30 or 50 min) and a vitrification solution containing 3 M cryoprotectant and 0.5 M sucrose (5, 10, 20, or 30 min). Embryonic cells that were pretreated for 30 min and vitrified for 20 min with ethylene glycol had the greatest rate of survival of embryonic cells (68.6%; P < 0.01), an optimal highest percentage of viable PGC (73.8 to 74.9%; P < 0.05), and no evidence of ice formation after thawing. The vitrified/thawed PGC were transplanted into blastula-stage embryos from goldfish (Carassius auratus). The PGC maintained their motility and moved to the gonadal ridge of the host embryo. Thus, the combination of vitrification and transplantation to produce germ-line chimeras is a powerful tool for the artificial production of next-generation offspring.     

Assessment of ovulation of common carp (Cyprinus carpio) females selected for induced spawning on the basis of external morphological characteristics

Secondary sexual characteristics such as softening and rounding of the abdomen as well as reddening and protrusion of the anal papilla and vent can be of help to breeders in selecting common carp (Cyprinus carpio) females prepared for propagation. To assess the reliability of this method, long-term data obtained on induced spawning of common carp at a large-scale fish hatchery were evaluated. The average spawning ratio of 2,620 females receiving hormonal injections was 79.8%. The average pseudogonadosomatic index (PGSI) calculated from data on the egg production of 2,086 females was 16.3 +/- 5.87% (mean +/- SD) for the same period. There was a correlation between fish weight and the time of induction determined by the breeder on the basis of external morphological characteristics. The similarity of the responses of females, including both spawning ratio and PGSI, among the different weight categories proved the reliability of this method for identification.