Gonadotropin response to GnRH during sexual ontogeny in the common carp, Cyprinus carpio

This study was designed to reveal whether gonadotropic response to GnRH in the common carp (Cyprinus carpio) changes during sexual ontogeny and whether the response of FSHbeta and LHbeta subunits is uniform or differential. The study comprised fish at the following stages: juveniles (4-month-old females with primary oocytes and early spermatogenic males); maturing (9-month-old previtellogenic females and advanced spermatogenic males); and mature (16-month-old postvitellogenic females and spermiating males). Fish were injected with superactive salmon GnRH analogue (sGnRHa; 25 microg/kg) and blood was sampled 6, 12 and 24 h later for cGtH (LH) and sex steroid levels. Pituitaries were taken for determination of FSHbeta and LHbeta mRNA levels by slot-blot hybridization and for cGTH content in the same glands by radioimmunoassay (RIA). Values were compared with the levels prior to sGnRHa administration and with control fish sampled at the same intervals. Juvenile fish did not respond at all to sGnRHa. In maturing females, FSHbeta mRNA increased by >300%, while that of LHbeta increased by 200%. In maturing males, FSHbeta mRNA did not change and only a slight increase occurred in that of LHbeta. In 16-month-old postvitellogenic females, there was no response of FSHbeta mRNA, while that of LHbeta dramatically increased. In spermiating males of the same age, mRNA of both FSHbeta and LHbeta increased following sGnRHa injection. Immunoreactive cGtH was present in the pituitary and plasma of all fish examined, but in juveniles it did not change following sGnRHa injection. In maturing and mature fish of both genders, sGnRHa administration was followed by a marked increase in circulating cGtH, concomitant with a decrease in its pituitary content, indicating the limited amount of the hormone stored in the gland. In conclusion, the response of the gonadotropin subunit mRNAs in the common carp was found to be differential and dependent on the gender and the phase of sexual ontogeny.     

Homologous radioimmunoassay and radioreceptorassay of gonadotropic hormone for an Indian carp Catla catla

A homologous radioimmunoassay (RIA) for Indian carp (C. catla) gonadotropin (GtH) was developed by using purified Catla GtH and its specific antiserum. In Ouchterlony agar diffusion, antisera raised against purified Catla GtH (cGtH), showed clear crossreaction. Catla-anti GtH (anti-cGtH) did not cross-reacted with Catla TSH enriched fraction. Immunocrossreaction was further confirmed with competitive binding inhibition studies where displacement of radiolabelled cGtH was precisely linear against increasing concentrations of cGtH, hence this was later used as standard curve for RIA. Competitive binding inhibition was also observed with purified murrel GtH, silver carp GtH and Cyprinus GtH, using varied doses. Both murrel and silver carp GtH showed clear parallelism with cGtH, while Cyprinus GtH inhibition slope was less steeper. Mammalian GtHs (hCG, oLH, oFSH), bTSH and bPRL had no crossreaction with anti-cGtH. Radioreceptorassay (RRA) for cGtH was developed by preparing plasma membranes from Catla oocytes. Binding of 125I-cGtH to oocyte plasma membranes showed saturability with high affinity (Ka = 0.11 X 10(13)M-1) and low capacity (17 fmol/mg protein). Displacement of 125I-cGtH in receptorassay by cold cGtH was linear and therefore served as standard curve. The interassay and intrassay variability in RIA was 7.9% and 3% while that of RRA was 5% and 3% respectively. Sensitivity of RIA was in the picogram level whereas it was in nanogram level by RRA. Determination of carp pituitary and serum GtH content by RIA and RRA showed the consistency, precision and validity of these assays. Although RRA was comparatively less sensitive than RIA, it was convenient, quick and less expensive.     

饲料营养成分对草鱼,团头鲂和青鱼鱼种阶段的饲料可消化能值的影响

为了探讨饲料可消化能值同饲料营养成分之间的关系,用Ｃｒ２Ｏ３作指示物,分别测定了鱼粉和大豆粕等饲料原料的草鱼（Ｃｔｅｎｏｐｈａｒｙｎｇｏｄｏｎ　ｉｄｅｌｌａ）团头鲂（Ｍｅｇａｌｏｂｒａｍａ ａｍｂｌｙｏｃｅｐｈａｌａ　Ｙｉｈ）青鱼（Ｍｙｌｏｐａｒｙｎｇｏｄｏｎ　ｐｉｃｅｕｓ）鱼种饲料的可消化能,用微机计算分析测试结果,发现饲料可消化能值随饲料蛋白质和／或脂肪食量增加而增加;随饲料无氮浸出物和／或纤维含量增加而降低。同时,“优选”出了有一定实用价值的估算草鱼、团头鲂和青鱼鱼种饲料可消化能值的回归方程。     

A quantitative study of testicular germ cell populations in masculinized neomale common carp (Cyprinus carpio L.)

The main objective of the present study was to investigate the effects of 17 alpha-methyltestosterone treatment upon the testicular germ cells of gynogenetic masculinized neomale common carp (Cyprinus carpio L.) in comparison with diploid carp. Gynogenetic common carp progeny (mean body weight, BW, 2.6+/-0.3g; mean total length, 10.4+/-0.5 cm) were treated for a period of 40 days with 17 alpha-methyltestosterone (MT) at a dose of 100mg kg(-1). The oral administration of MT resulted in 61.5-100% of fish exhibiting male gonads. The masculinized neomales exhibited reduced (P<0.05) body weight (BW=22.9+/-0.8) but significantly increased (P<0.05) mean testis weight (2.1+/-0.3) and mean gonadosomatic index (GSI=9.5+/-0.2%) in comparison with fish not treated with MT (BW 54.8+/-1.3; GSI=0.61%). Furthermore, treatment with MT also resulted in an increased number of fish exhibiting abnormal gonads. However, neomales did not exhibit abnormalities in the development of sperm ducts. MT treatment significantly increased germ cell volume, nuclear diameter, nuclear volume and cyst volume (P<0.01 in all cases) in MT-treated fish compared to untreated fish. The area occupied by seminiferous tubules, the number of Sertoli cells and germ cells per cyst, and the number of Leydig cells were significantly (P<0.05) greater in fish treated with MT. The carp neomales exhibited approximately 20-60% more Sertoli cells per cyst (P<0.05). Leydig cell nuclear volume and Leydig cell individual volume were significantly reduced in MT-treated groups (P<0.05) compared with untreated groups. In conclusion, our study strongly suggests that the abnormal gonadal structure evident in masculinized neomales could be explained by a combination of MT-induced genetic (homozygosity) and anabolic effects (upon germ and somatic cells).     

IGF—ImRNA在不同年龄鲤表达的差异和LHRH—A对IGF—ImRNA表达的影响

利用RNA酶保护法对7月龄性未成熟幼鲤和2龄性成熟鲤组织胰岛素样生长因子-I（IGF-I）mRNA的表达水平进行测定,结果表明成鱼肝和肾脏组织IGF-ImRNA的丰度显著高于幼鱼,对鲤成鱼和幼鱼腹腔注射促性腺激素释放激素类似物（LHRH-A,D-Ala^6-Pro^9-NEt-LHRH)使血清生长激素（GH）水平和肝组织IGF-ImRNA水平都显著升高,而成鱼生殖腺IGF-ImRNA的丰度比对照组显著增加,研究结果提示鲤在不同发育阶段肝组织IGF-ImRNA的丰度比对照组显著增加,研究结果提示鲤在不同发育阶段肝组织IGF-ImRNA的表达存在差别,其中2龄成鱼大于7月龄幼鱼;LHRH-A可能通过刺激垂体GH的释放间接促进肝组织IGF-ImRNA的表达,亦可能通过某种未知途径刺激生殖腺IGF-ImRNA的表达。     

Dietary fish oils modify adipocyte structure and function.

Dietary fish oils, enriched with omega-3 fatty acids (e.g., MaxEPA fish oil), inhibit lipogenesis and have a marked hypotriglyceridemic effect in man and experimental animals. Dietary omega-3 fatty acids also reduce adipose tissue trophic growth in rats. To understand the metabolic basis for this, we measured the effect of fish oil feeding upon rat plasma triglyceride concentration, fat pad mass, fat cell size, fat cell lipolysis, as well as lipoprotein binding to adipocyte plasma membranes. In adolescent (250 g) male Wistar rats fed 20% (w/w) fish oil supplemented diets for 3 weeks, plasma triglyceride levels and epididymal and perirenal fat pad mass were significantly (P less than 0.005) reduced compared to pair-fed controls given 20% lard diets. These differences in fat pad mass between the diets were greater than differences in whole animal mass or in the mass of livers, testes, kidneys, spleens, or hearts. Isoproterenol-stimulated lipolysis was significantly (P less than 0.005) higher in fish oil fed rats than in pair-fed controls. In young (100 g) rats plasma triglyceride levels were 10 times lower in the fish oil fed group after 5 weeks as compared to the lard-fed controls. This was accompanied by a reduction in epididymal and perirenal fat pad mass as well as a 2-3-fold decrease in adipocyte volumes; there was no significant difference between the two groups in fat cell number in each region. Plasma membranes of epididymal adipocytes from fish oil fed rats bound significantly (P less than 0.001) less HDL1 than the lard-fed rats, possibly as a result of a reduction in fat cell size and/or alteration of plasma membrane structure. Thus in both young and old rats, the reduction in plasma triglyceride concentration in conjunction with increased hormone-stimulated lipolysis may explain in part the selective reduction in adipose tissue trophic growth accompanying fish oil consumption.     

Effects of various hormones on the sexual maturity of European eel (Anguilla anguilla L.) females from farm and lakes

Previously described and alternative methods of the induction of sexual maturation in the European eel were investigated. Weekly administrations of a gonadoliberin agonist (nnRH-A=D-Phe6-GnRH-Ea) did not induce statistically significant effect on the gonads of treated eels in none of the dosages used (0.1 microg and 10 microg/fish). Carp pituitary extract and carp pituitary extract together with a dopamine antagonist caused considerable external changes (increase in eye size) and significant gonadal development in two treatment groups: wild and cultivated stocks. The induction of the ovulation by double amount of CP and gonadoliberin agonist with dopamine antagonist mixture was not successful in a wild stock. Fertilisation of stripped eggs of farm eel was attempted unsuccessfully in, due to low egg quality. An advanced phase of the sexual maturation process could be induced in specimen infected by Anguillicola crassus indicating, that nematode infection is not a limiting factor in the artificial propagation of the European eel.     

[125I] gonadotropin binding to the ovary of an Indian major carp,Catla catla,at different stages of reproductive cycle

At different stages of the annual reproductive cycle ofCatla catla, a major Indian carp, specific binding of gonadotropic hormone to the plasma membrane receptors was demonstrated. Maximum specific binding of [¹²⁵I]Catla gonadotropic hormone was obtained at 30‡C and pH 7.5 during 2 h of incubation.Catla gonadotropic hormone binding was saturable with high affinity. Competitive inhibition experiment showed that binding site was specifically occupied by piscine gonadotropic hormone,Catla gonadotropic hormone and murrel gonadotropic hormone, human chorionic gonadotropin was a weak competitor while bovine thyroid stimulating hormone, bovine prolactin and ovine follicle stimulating hormone had no effect. Scatchard analysis ofCatla gonadotropic hormone binding to the plasma membrane preparation from the carp oocytes of different reproductive stages showed that the range of dissociation constant(K _d ) varied from 0.78 to 0.97 x 10^-10 M. However, maximum binding capacity (B-max) varied remarkably between the different stages of reproductive cycle, it was 6.11 ± 0.36 fmol/mg protein in the preparatory stage which increased to about three-fold in prespawning stage of reproductive cycle (17.0 ± 0.29 fmol/mg protein) and spawning (18.7 ± 0.17 fmol/mg protein) and lowest in postspawning stage of reproductive cycle (5.28 ± 0.28 fmol/mg protein). Fluctuation in the number of gonadotropic hormone binding site at different stages of annual reproductive cycle was found to be coincided well with the pattern of ovarian steroidogenesis in response toCatla gonadotropic hormone as determined by the formation of progesterone from pregnenolone.     

Sympathetic denervation does not prevent a reduction in fat pad size of rats or mice treated with peripherally administered leptin

Leptin increases sympathetic nervous system (SNS) activity in brown adipose tissue and renal nerves. Experiments described here tested whether SNS innervation is required for peripheral, physiological concentrations of leptin to reduce body fat. In experiment 1, one epididymal (EPI) fat pad was sympathectomized by local injection of 6-hydroxydopamine (6OHDA) in C57BL/6 mice that were then infused for 13 days with PBS or 10 microg leptin/day from an intraperitoneal miniosmotic pump. Surprisingly, EPI denervation increased total body fat of PBS-infused mice but leptin decreased the size of both injected and noninjected EPI pads in 6OHDA mice. Experiment 2 was identical except for the use of male Sprague-Dawley rats that were infused with 50 microg leptin/day. Leptin had little effect on EPI weight or norepinephrine (NE) content, but denervation of one EPI pad decreased the effect of leptin on intact EPI, inguinal and retroperitoneal (RP) fat and increased the size of the mesenteric fat pad. Experiment 3 included groups in which either one EPI or one RP pad was denervated. RP denervation reduced RP NE content but did not prevent a leptin-induced reduction in fat pad mass. Therefore, the SNS is not required for low doses of leptin to reduce body fat. EPI denervation significantly increased adipocyte number in contralateral EPI and RP fat pads and this was prevented by leptin. These changes in intact pads of rats with one denervated fat pad imply communication between fat depots and suggest that both leptin and the SNS regulate the size of individual depots.     

Feeding behavior of black carp Mylopharyngodon piceus (Pisces: Cyprinidae) on fry of other fish species and trematode transmitting snail species

Fish raised in aquaculture ponds may get infected with fishborne zoonotic trematodes (FZT) during the nursing stage. Freshwater snails serve as intermediate hosts for FZT and we wanted to explore the possibility of controlling snails by stocking nursery ponds with a few juvenile specimens of the mollusc-eating fish, black carp (Mylopharyngodon piceus). Obviously, the risk that black carp might also prey on the juvenile fishes in nursery ponds should first be assessed. Laboratory trials showed that all size classes of juvenile black carp consumed fry of common carp (Cyprinus carpio) even when offered snails as food; the odds of survival of fry from tanks with medium sized and large black carp was 5.6% and 39.9%, respectively of that of fry in tanks with small sized black carp. Since the large black carp also consumed fewer snails than medium sized fish, we believe that large specimens were stressed in the experimental aquaria. Under semi-field conditions, presence of the black carp had no effect on survival of fry of Oreochromis niloticus and C. carpio both in the absence and presence of snails as alternative food. The black carp consumed most snails offered with the exception of some of the large snails. We conclude that under field conditions, predation by black carp on fish fry is minimal and field trials in nursery ponds are warranted. Due to the risks that black carp pose to native imperiled snails and other molluscs, trials should be restricted to ponds within the fish’s native or existing range.     

Gonad development and filleting yield of common carp Cyprinus carpio L. reared in ponds in Eastern France

The processing of 232 common carp Cyprinus carpio made it possible to study the correlation between gonad maturation and filleting yield. The study was undertaken between April and August 1992 and was carried out on 4-year-old carp cultured in extensive ponds. Morphological parameters were recorded and relations between total weight and length, biomass of fillet, and biomass of gonad were determined. The histology of male and female gonads was examined in relation to the gonadosomatic index (GSI: up to 16% for females and less than 11% for males) and condition indices (total K or somatic K1). Filleting yields (32–36% of the total weight) decreased for females just prior to spawning and corresponded to a decline of K1. A rematuration phenomenon was observed, although females weighing less than 1200g were still immature. We suggest avoiding the filleting of large females before spawning and suggest estimating the date of appearance of this stage by referring to the positive correlation established in this paper between the number of degree-days and total fish weight.     

The relation between pituitary gland and thyroid growth during the lifespan of the annual fish Cynolebias whitei and Nothobranchius korthausae: gonadotropic and thyrotropic cells

Summary In the annual cyprinodont Cynolebias whitei the cell types responsible for the increase of pituitary growth at the onset of maturation and for pituitary hyperplasia in old specimens were identified as gonadotropic cells and thyrotropic cells, respectively. The gonadotropic cells showed a high affinity to anti-carp -GTH serum, both at light- and electron-microscopical levels. The allometric relation of total gonadotropic cell volume to body length, determined for fish from six weeks up to six months of age, showed no inflections. Therefore pituitary growth in maturing fish may be partly a result of proliferation of gonadotropes, although gonadotropic cells do not contribute to pituitary hyperplasia in old fish. Thyrotropic cells showed a weak affinity to anti-carp -GTH serum at light-microscopical level. Under the electron microscope thyrotropic cells displayed signs of activation in maturing fish and signs of proliferation in old fish. The allometric relation of thyroid gland volume to body length paralleled that of pituitary volume to body length. Histologically the thyroid gland showed signs of inactivity in adult fish and of hyperplasia in old fish. The possibility, that gonadal maturation, pituitary thyrotropic activity, and growth of the thyroid in maturing fish are related through the inhibitory action of gonadal steroids on thyroid hormone release, is discussed. Pituitary hyperplasia in old fish is the result of proliferation of thyrotropic cells. Similar hyperplasia of pituiary and thyroid glands was observed in old Nothobranchius korthausae.     

Potential problems with removing scales before surgical transmitter implantation

Thirty one mirror carp Cyprinus carpio were implanted with telemetry transmitters. Several large scales were removed from the incision area of two mirror carp before implantation of transmitters. When the fish were recaptured after 4 months, the mirror carp from which scales were removed showed severe tissue necrosis at the incision site whereas intact fish did not.     

Genetic differences between the Chinese and the European races of the common carp III. Gonad abnormalities in hybrids

Abnormalities in the gonads of carp, Cyprinus carpio L., were present in a relatively high proportion of hybrids derived from the Chinese and European strains. The aberrations involved were the absence or marked reduction of the gonads in mature fish.     

Development of hypophysis and differentiation of its secretory cells at the early postembryonic development of silver carp Hypophthalmichthys molitrix (cyprinidae)

Silver carp Hypophthalmichthys molitrix hatches from the envelope at phase 19 a day after fecundation (water temperature 26–28°C) with body length 5.2 mm. The formation of hypophysis begins 7–8 h after hatching of the embryo. The first chromophilic cells (acidophiles in adenohypophysis) appear at the age of 9 days; the first basophilic cells (thyrotropocytes) at the age of 17–20 days, when juveniles are at stage III. The gonadotropic function is formed in the ontogenesis of silver carp at the age of 40–45 days, i.e., almost a month after the appearance of elementary sexual cells in the gonads. The differentiation of gonadotropocytes in mesoadenohypophysis to two functionally active types of cells (central and peripheral) is also completed by this time.     

Seasonal changes in the biochemical composition of the freshwater murrel,Ophicephalus punctatus Bloch

Summary Seasonal variations in various biochemical constituents in the tissues of Ophicephalus punctatus showed that. the fat content in the muscle was highest during November–January and lowest during June–October. Liver contained more fat than the muscle and its seasonal cycle was marked by two well defined peaks. Changes in the ovarian fat were also well defined seasonally. The moisture content in all the tissues varied inversely with the fat. Protein values in different tissues did not show as much fluctuations as fat. In muscle, protein remained high throughout the year except in January. In liver and gonads very low protein values were recorded during the winter months. Seasonal changes in the ash content were not very well marked in any of the three tissues.The seasonal variations in all the biochemical constituents seem related to the feeding intensity of the fish and the maturation of its gonads. Maximum and minimum values of fat fell in accordance with the high and low food intake. In the pre-spawning phase, more fat was stored in the muscle at first but subsequently the fat was mobilized in the liver and finally in the gonads. Peak ripeness was associated with an accumulation of greater protein and fat in the gonads (ovaries). During the spawning months the various constituents were depleted and the post-spawning phase of the fish was marked by minimal values of most of the biochemical constituents, excepting moisture and ash.     

Seasonal changes in the chemical composition of fish body and nutritional value of the muscle of the pollan (Coregonus pollan Thompson) from Lough Neagh,Northern Ireland

Studies on the chemical composition of adult pollan were carried out monthly from January to December 1977. Sex, weight of scaled fillet, and gonads were examined in 184 fish. Chemical composition which includes water, fat and ash content was analysed in the whole fish, muscle and gonads. This paper attempts to account for the fat and protein flow within fish of both sexes during gonad maturation. Extreme differences between males and females have been recorded in the fat deposition process during summer and spawning period. In spite of our fragmentary knowledge of spawning behaviour in pollan, an explanation is offered for these phenomena. The changes in fat content of female gonads were followed closely throughout the year and the cost of egg production was estimated as a percentage of the amount of fat deposited in the body. Since a similar growth pattern was found for both sexes of pollan, differences in feeding intensity might account for the difference in energy expenditure. Amino acid content was analysed in muscle and gonads and an Essential Amino Acid Index (EAAI) was calculated. There were some changes in EAAI which depended on sex and season. Finally, a new method for evaluation of muscle production expressed as fat and protein is used to determine the most suitable time for fish harvesting.     

综合养鱼高产池塘的溶氧变化周期

根据1979—1984年对主养鲢、鳙、非鲫,主养青、草鱼,主养青鱼3种养殖结构类型高产鱼池溶氧变化周期的系统研究,揭示了高产养鱼池塘溶氧的昼夜、垂直、水平变化及季节变化周期,分析了光合作用、呼吸作用、扩散作用在高产养鱼池塘溶氧动力学上的地位和作用。对高产养鱼池塘的溶氧收入、支出及平衡情况进行了定量研究,在主要饲养季节,测得晴到多云天、晴天的溶氧来源分别为:浮游植物光合产氧占86.0和95.3%,大气扩散溶入占14.0和4.7%;氧的消耗分别为:“水呼吸”消耗氧占72.0和72.0%,鱼类呼吸消耗氧占22.0和13.1%,淤泥中生物呼吸消耗氧占2.9、5.5%,扩散逸出占3.1、8.8%。文中还对溶氧变化周期与养鱼池塘管理的环境控制、结构控制之间的关系进行了分析。