Steroid hormone secretion by testicular tissue from African catfish,Clarias gariepinus,in primary culture: identification and quantification by gas chromatography — mass spectrometry

With the final aim of identifying the testicular steroids involved in the feedback mechanism of the hypothalamus-pituitary-gonad axis, steroid secretion by the testis of African catfish, Clarias gariepinus, was studied in vitro, by means of gas chromatography followed by mass spectrometry. Testicular fragments of sexually mature catfish raised in captivity were incubated in L-15 medium with and without catfish pituitary extract (cfPE). Without adding cfPE, 22 steroids could be identified, amongst which 11β-hydroxyandrostenedione, 11β-hydroxytestosterone, 11-ketotestosterone and 11-ketoandrostenedione were dominating. After incubation in the presence of cfPE, the concentrations of the four 11-oxygenated steroids were increased about 4-fold. The amounts of pregnane derivatives in the incubation medium showed the largest increases in the presence of cfPE. 5β-Pregnane-triol levels, for example, were 60-fold higher than in the medium from control incubations. The secretion of 5α- and 5β-androstanes was also stimulated by cfPE. The stimulation was not equal for all steroids, indicating that cfPE not only stimulates total steroidogenesis by increasing the availability of cholesterol, but also by influencing specific steroid converting enzyme activities. Part of this work was presented at the IV^th International Symposium on the Reproductive Physiology of Fish, 7–12 July 1991, Norwich, U.K.     

Steroidogenesis during induced oocyte maturation and ovulation in the African catfish,Clarias gariepinus

Changes in ovarian steroidogenesis accompanying oocyte maturation and ovulation were studied in the African catfish,Clarias gariepinus. Laboratory-reared females with postvitellogenic ovaries were treated with pimozide and LHRH-analogue. The plasma gonadotropin levels were determined by means of a homologous radioimmunoassay, the condition of the ovaries was studied by histological examination of the follicles, and the steroidogenetic capacity of the ovaries was analyzed byin vitro incubation of tissue fragments for 3 h with [³H]-pregnenolone and [³H]androstenedione as precursors. Data were collected at regular intervals between 0 and 16 h after pimozide-LHRH analogue administration.Until 4 h after the beginning of the experiments the plasma gonadotropin levels did not rise above 25 ng/ml, the ovaries remained in the stage of postvitellogenesis, and testosterone was the main end product of steroidogenesis. Four hours later the gonadotropin concentration in the blood had risen to more than 150 ng/ml, and the ovaries had entered the stage of germinal vesicle migration. At the same time steroidogenesis shifted towards the production of 17,20-dihydroxy-4-pregnen-3-one, 5-pregnane-3, 17-diol-20-one, 5-pregnane-3,6,17-triol-20-one, 5-pregnane-3,17,20-triol and 5-pregnane-3,6,17,20-tetrol. During the subsequent stage of germinal vesicle breakdown the plasma gonadotropin level remained high, and the synthesis of the C₂₁-steroids showed a further increase. Simultaneously, the production of some C₁₉-steroid glucuronides was enhanced. The preovulation and especially the postovulation stages were accompanied by a gradual decrease in steroidogenic capacity of the ovaries, even though the plasma gonadotropin level remained high. It is concluded that the prematuration surge of gonadotropin influences the activity of enzymes involved in steroidogenesis, leading to a reduced C_17,20-lyase and to an augmented activity of the enzymes 20-hydroxysteroid dehydrogenase (HSD), 5-reductase, 3-HSD, 6-hydroxylase and UDP-glucuronosyltransferase. During ovulation the activity of all steroidogenic enzymes, including such key enzymes as 3-HSD and 17-hydroxylase, gradually decreases.Not only 17,20-dihydroxy-4-pregnen-3-one, but also the 5-reduced pregnanes may be involved in inducing oocyte maturation and/or ovulation. The very polar triol and tetrol products may function, together with the steroid glucuronides as sex pheromones.A preliminary account of these results was presented at the XIII Conference of European Comparative Endocrinologists, Belgrade, September 7–12, 1986     

Mismatch between patterns of circulating and testicular androgens in African catfish, Clarias gariepinus

11-Ketotestosterone (11-KT) is an important plasma androgen in male African catfish. The quantitatively predominating androgen produced by the testis, however, is 11-hydroxyandrostenedione (OHA). Our working hypothesis to explain this mismatch assumed that OHA is converted to 11-KT at extratesticular sites. First, we examined the in vivo metabolism of [³H]-OHA in mature males after sham-operation or removal of either the testes (TC), the seminal vesicles (SVC), or both (TSVC) by analysing the pattern of circulating [³H]-androgens two hours after intravenous injection of [³H]-OHA. [³H]-OHA was converted to [³H]-11-KT to the same extent in all groups, indicating that neither ablation of testes nor of seminal vesicles, or both attenuates this conversion. We then examined the in vitro metabolism of [³H]-OHA by several types of tissues. Liver and seminal vesicle tissue were found to produce significant amounts of [³H]-11-KT. The conversion capacity in vivo was assessed by injecting TSVC-castrated males with increasing doses of radioinert OHA, followed by the quantification of OHA and 11-KT plasma levels. Saturation of the conversion capacity was not reached but the 11-KT production capacity is at least 80 ng per ml of plasma per hour. Moreover, liver fragments were tested for their OHA to 11-KT conversion capacity in vitro using increasing concentrations of radioinert OHA. The 11-KT producing increased with time and OHA concentration. The production rate was 90 pg 11-KT mg^-1 liver h^-1. Considering the results of the surgical experiments and the fact that the total hepatic mass by far exceeds that of the seminal vesicles, we conclude that the hepatic conversion is of primary relevance in vivo.     

Plasma profiles of fourteen ovarian steroids before,during and after ovulation in African catfish,Clarias gariepinus,determined by gas chromatography and mass spectrometry

The plasma concentrations of fourteen ovarian steroids were measured in postvitellogenic African catfish,Clarias gariepinus, which had been injected with pimozide and LHRHa. Postvitellogenesis persisted for at least four hours after pimozide and LHRHa administration. During this stage a limited rise in the plasma gonadotropin (GTH) level was accompanied by an increase in the testosterone concentration. The estradiol level was high and remained high except for a passing drop during the stage of germinal vesicle migration. At the stage of germinal vesicle migration a strong increase in the plasma GTH level coincided with a maximum in the testosterone concentration and a concomitant increase in the levels of 17,20-dihydroxy-4-pregnen-3-one and of five 5-reduced pregnanes. During germinal vesicle breakdown the GTH concentration remained high, the plasma level of 17-hydroxyprogesterone tended to increase, and the levels of 5-pregnane-3, 17-diol-20-one, 5-pregnane-3,17,20-triol and 5-pregnane-3,17,20-triol reached a maximum. At pre-ovulation the GTH concentration did not change, and peak levels were reached of 17,20-dihydroxy-4-pregnen-3-one and 5-pregnane-3,6,17-triol-20-one. Shortly after ovulation the GTH concentration slightly decreased together with a sharp decline in the concentrations of 17,20-dihydroxy-4-pregnen-3-one and the 5-reduced steroids, with the exception of 5-pregnane-3,17,20-triol, 5-pregnane-3,6,17,20-tetrol and 5-dihydrotestosterone. The plasma concentrations of androstenedione, estrone, etiocholanolone and 5-androstane-3,17-diol showed marginal fluctuations during oocyte maturation and ovulation. Apart from 17,20-dihydroxy-4-pregnen-3-one, the 5-reduced pregnanes might be candidates for the function of oocyte maturation inducing hormone inC. gariepinus.     

Catecholestrogens inhibit dopamine methylation in the gonadotrops of the African catfish,Clarias gariepinus

Isolated gonadotrops of the African catfish,Clarias gariepinus, were incubated with dopamine (DA) and/or catecholestrone and the activity of the enzyme catechol-O-methyltransferase (COMT) was determined by measuring the methylated products. From the apparent Km values for DA and catecholestrone of 0.4–1.3 M and 17.9–25.2 M respectively, it was concluded that catecholestrone is a better substrate for the enzyme COMT, compared to DA. Moreover, the methylation of DA is inhibited by comparatively low concentrations of catecholestrone.     

In vivo activity of native GnRHs and their analogues on ovulation in the African catfish, Clarias gariepinus (Burchell)

Four distinct forms of native gonadotropin‐releasing hormone (GnRH) and two newly designed analogues were tested for their in vivo activity to induce ovulation in African catfish. The effects of these peptides on ovulatory parameters were compared with those of carp pituitary and [d ‐Ala⁶, Pro⁹‐NEt]‐mammalian GnRH analogue (mGnRHa), two tested ovulation‐inducing agents in African catfish. Assessment of ovulation was carried out by determining the ovulation ratio and the relative quantity of egg produced. From the results of the experiments, the order of potency of the native GnRH peptides is summarized as chicken GnRH‐II (cGnRH‐II) >salmon GnRH (sGnRH) >mammalian GnRH >chicken GnRH‐I (cGnRH‐I). Chicken GnRH‐II was as potent as mGnRHa while cGnRH‐I was totally ineffective. The new d ‐Orn⁶‐cGnRH‐II and d ‐Orn⁶‐sGnRH with a substitution at position 6 with d ‐isomer residue were as potent as the most extensively used mGnRHa, indicating that the position 6 modification might be more crucial than the substitution at the C‐terminal. On the basis of our results, the potential use and incorporation of cGnRH‐II and sGnRH for the development of more generic spawning induction therapies are suggested.     

Optimum histidine requirement of fry African catfish, Clarias gariepinus (Burchell)

Dietary histidine requirement of fry African catfish, Clarias gariepinus (2.57 ± 0.02 cm; 0.22 ± 0.03 g) was quantified by feeding casein–gelatin-based isonitrogenous (40% crude protein) and isocaloric (17.90 kJ g⁻¹ gross energy) amino acid test diets with graded levels of histidine (0.25%, 0.30%, 0.35%, 0.40%, 0.45% and 0.50% dry diet) in eighteen 80 L indoor circular aqua-coloured troughs provided with the flow-through system for 12 weeks. Maximum absolute weight gain (2.66), best feed conversion ratio (1.29), highest protein efficiency ratio (1.94), protein retention efficiency (34%) and energy retention efficiency (70.4%) were achieved at 0.40% dietary histidine. Broken-line and non-linear regression models were adopted to assess dietary histidine requirement for C. gariepinus . When analysed using broken-line regression model these parameters were also best at 0.40% dietary histidine corresponding to 1.0% protein, respectively, whereas using second-degree polynomial regression analysis, histidine requirement was obtained at 0.42%, 0.41%, 0.40%, 0.41% and 0.41% of dry diet, corresponding to 1.05%, 1.02%, 1.0%, 1.02% and 1.02% protein respectively. Based on the broken-line and second-degree polynomial regression analyses of the growth and nutrient retention data, optimum histidine requirement of fry C. gariepinus was found to be in the range of 0.40–0.42% dry diet, corresponding to 1.0–1.05% of dietary protein.     

The feedback regulation of pituitary GTH-II secretion in male African catfish (Clarias gariepinus): Participation of 11-ketotestosterone

11-ketotestosterone (OT) is a typical androgen of male teleost fish, but information on the question if it is involved in the feedback regulation of pituitary gonadotropin II (GTH-II) secretion is controversial. We have therefore studied the effects of OT on gonadotropin releasing-hormone (GnRH) stimulated GTH-II secretion in male African catfish Clarias gariepinus). In vivo experiments were carried out with intact and castrated fish. OT plasma levels were increased by implantation of silastic capsules containing 11-ketoandrostenedione (OA) which is converted to OT in both intact and castrated fish. When intact males received OA- or blank-capsules, treatment with salmon gonadotropin releasing-hormone analogue (Des-Gly¹⁰-D-Arg⁶-sGnRH-NEt; 0.2 μg sGnRHa/kg body weight) elevated the plasma GTH-11 levels in both groups. However, the levels were about 2 times higher in blank- than in OA-implanted fish. When castrated fish received either blank-or OA-capsules, sGnRHa treatment led to plasma GTH levels significantly higher than in sham-operated fish. However, there was no difference between the blank- or OA-implanted castrates, though OA implantation led to a restoration of OT plasma levels. This suggests that replacement ofOT is insufficient to reverse castration-induced effects. In vitro experiments were carried out with pituitary tissue fragments using a static culture system. The tissue remained sensitive to sGnRHa (5 × 10^?9M) for 4 days after the beginning of incubation. Preincubation of pituitary tissue for 24 hours with 25 ng OT/ml medium (80 nM) completely abolished the stimulatory effect of sGnRHa on GTH-II secretion. Tritiated OT was not metabolized by pituitary tissue during 6 hours of incubation. We conclude that 11-ketotestosterone, a quantitatively prominent and non-aromatizeable circulating androgen participates, at least in part by direct action on the pituitary, in the negative feedback regulation of GnRH-stimulated GTH-II secretion in male African catfish.     

Nicotinic acid supplementation of diets for the African catfish, Clarias gariepinus (Burchell)

Semi-purified diets containing 39% crude protein and 5% lipid were used to identify the qualitative requirement of African cattish, Clarias gariepinus (Burchell), for niacin and to characterize the pathologies associated with a deficiency of this vitamin. After 48 days of feeding, C. gariepinus supplied with the unsupplemented diet had developed severe deficiency symptoms and were subsequently withdrawn from the growth study. Niacin deficiency was characterized by feed refusal, listlessness, weight loss, poor feed utilization and high mortality. The skin overlaying the lateral line of the deficient fish became haemorrhagic and this clinical sign was accompanied by severe anaemia. After 126 days of feeding, fish fed diets containing 17.0 mg niacin kg^?1 had also developed a dermopathy. but without anaemia or high mortality. The feeding of diets containing less than 33.1 mg niacin kg^?1 resulted in suboptimal feed efficiency and poor protein utilisation. Allometric analysis of proximate composition indicated that carcass moisture, protein and ash were influenced by fish size, and not by dietary niacin content. However, significantly more lipid per unit of weight gain was deposited in the carcasses of fish fed the unsupplemented diet than in fish fed diets containing 17.0mgkg^?1. The indicators used in the present study could not be applied to accurately determine a value for niacin requirement. However, until a more accurate assessment is performed, it is recommended that diets for C. gariepinus contain not less than 33.1 mg nicotinic acid kg^?1 feed.     

Characterization of the receptor for gonadotropin-releasing hormone in the pituitary of the African catfish,Clarias gariepinus

Receptors for gonadotropin-releasing hormone (GnRH) were characterized using a radioligand prepared from a superactive analog of salmon GnRH (sGnRH), D-Arg⁶-Pro⁹-sGnRH-NEt (sGnRHa). Binding of¹²⁵I-sGnRHa to catfish pituitary membrane fractions reached equilibrium after 2 h incubation at 4°C. Displacement experiments with several GnRH analogs as well as other peptides, demonstrated the specificity of¹²⁵I-sGnRHa binding. Specific binding was enhanced in the presence of the cation chelator ethylene bis (oxyethylenenitrilo) tetra-acetic acid (EGTA), indicating an inhibitory effect of cations on GnRH-receptor binding. The binding of¹²⁵I-sGnRHa to pituitary membranes was found to be saturable at radioligand concentrations of 5 nM and above. A Scatchard analysis of the saturation data suggested the presence of a single class of high-affinity binding sites (Ka=0.901±0.06×10⁹M^–1, Bmax=1678±150 fmol/mg protein). A comparative study on¹²⁵I-sGnRHa binding to pituitary membrane fractions of male and female catfish, indicated that there were no differences in binding affinity and binding capacity between both sexes. The results demonstrate the presence of specific, saturable GnRH receptors in the African catfish pituitary.     

Gonadal steroidogenesis and the possible role of steroid glucuronides as sex pheromones in two species of teleosts

In general, female zebrafish,Brachydanio rerio, ovulate only in the presence of males. The stimulant must be pheromonal as even male holding water is capable of inducing ovulation. After ovulation the mating phase begins. During this phase the male follows the female and oviposition as well as fertilization takes place. Both the ovulation and the mating are controlled by pheromones synthesized by the gonads. Ovulation can be induced by testicular homogenates. After the lipid material has been extracted from the testicular homogenates, the remaining aqueous phase can still induce ovulation. However, when the aqueous phase is treated with the enzyme-glucuronidase, it loses the ability to induce ovulation. This is an indication that glucuronides, probably steroid glucuronides, are the compounds responsible.During the mating phase, ovulated female zebrafish become attractive to males. It was found that, after ovulation, ovarian extracts contain the compounds responsible for attracting males. The attractant consists of a mixture of steroid glucuronides.After incubation of the gonads with³H-precursors seven steroid glucuronides have been identified in the testis and five in the ovary.Under fish culture conditions the African catfish,Clarias gariepinus, can produce postivitellogenic oocytes throughout the year. However, in capitivity neither males nor females spawn. In female catfish maturation and ovulation can be induced by treatment with gonadotropins. It might be possible that, analogous to the zebrafish, some reproductive processes in the catfish have to be induced by pheromones. It has been demonstrated that pheromonal compounds released by the seminal vesicles are involved in the attraction of female conspecifics. The steroid glucuronide synthesizing capability of the testes and the seminal vesicles of the male catfish are examined, as well as that of the ovary before and after ovulation of the female catfish. Both testes and seminal vesicles appear to be capable of steroid biosynthesis but only the latter synthesizes steroid glucuronides. Six of these conjugates have been isolated and identified. In the female catfish the ovaries are capable of synthesizing seven steroid glucuronides, but only after ovulation.     

Effect of plant density in coupled aquaponics on the welfare status of African catfish,Clarias gariepinus

We investigated the effects of plant density on the welfare of African catfish, Clarias gariepinus, in coupled aquaponics over 85 days. The moderate density (mpd) of basil, Ocimum basilicum, was compared with the high density (hpd) and control (n = 0). The behavior was analyzed by visual and video observations, and after the application of induced stressors, skin injuries, blood glucose, lactate, and plasma cortisol responses were considered. The hpd fish showed the least activity (control: visual 77.8%, video 81.6%; mpd: 74.6%, 82.6%; hpd: 63.2% [p < 0.05], 78.8%). High agonistic behavior (control: 5, 131; mpd: 4, 57; hpd: 1, 45) and the highest number of injuries (control: 3.9; mpd: 2.9; hpd: 3.4) were observed in the control. Glucose and lactate levels did not differ significantly (control: 5.5, 2.6 mmol/L; mpd: 5.6, 2.7 mmol/L; hpd: 5.3, 2.6 mmol/L); however, cortisol levels did (control: 18.8 ng/mL, mpd: 19.9 ng/mL, hpd: 25.8 ng/mL). pH adjustment led to additional stress, resulting in temporal cortisol alterations. While in the control and mpd, low cortisol levels were followed by acute responses and downregulation, the hpd fish showed prior elevation and lagged an acute response. However, comparing injuries and behavioral patterns with control, aquaponics with high basil density influenced African catfish positively.     

Inherent variation in growth efficiency of African catfish Clarias gariepinus (Burchell, 1822) juveniles

Understanding the major causes of growth variation is crucial for the success of fish farming since its reduction contributes to maximize production efficiency, reduce food waste and improve water quality. The growth variation observed in aquaculture has been associated with the establishment of social hierarchies. However, some studies suggest that this variation may not be mainly a consequence of social hierarchies but mainly a result of inherent (genetic) differences. This study investigates the magnitude of individual responses, independently of group effects (fish housed individually), in growth efficiency and feeding behaviour of African catfish Clarias gariepinus (Burchell 1822). Despite the low variation in initial body weight (6.5%) and cumulative feed consumption (7.5%) over the experimental period, catfish exhibited high variation in final body weight (18.1%), specific growth rate (17.2%) and feed conversion ratio (27.9%), suggesting that individual variation in growth efficiency is important in determining growth rate. This individual variation may be related with individual differences in protein/fat deposition since faster growing fish deposited more protein and less fat than slower growing fish. Pronounced individual differences in feeding behaviour (reaction towards feed and time spent eating) were also observed and correlated with individual differences in growth efficiency. Fast eaters were the fast growers. We suggest that the growth variation observed in African catfish may be inherent and that the use of grading to increase uniformity should be further investigated.     

Purification of vitellogenin from the air breathing catfish, Clarias gariepinus

Vitellogenin (Vtg) is a female specific glycophospholipoprotein which can be induced both in male and female with estradiol and xeno-estrogens. The basic theme behind the purification of vitellogenin from the fish is to understand the evolutionary relationship and for the purification and characterization of the Vtg receptor. The male catfish, Clarias gariepinus was administrated with estradiol over a period of time for the synthesis of Vtg and the serum was collected. The Vtg was purified from the serum using a two step chromatographic technique. The serum was passed on to DEAE-ion exchange column and the protein was eluted using a salt gradient. The fractions containing the Vtg were pooled and passed onto a gel permeation chromatography column and the pure protein was obtained. The molecular weight is around 200 kDa on the SDS-PAGE and around 520 kDa on the native gel electrophoresis.     

GTH-cells in the pituitary of the African catfish,Clarias gariepinus,during gonadal maturation: an immuno-electron microscopical study

In an ultrastructural immunocytochemical study we investigated the development of the gonadotropic cells in the pituitary of two to six months old male African catfish in relation to testicular development. In this period, pituitary and testicular tissue samples were collected on five occasions (groups I–V). Blood samples could only be taken from the fish in groups III–V. The testicular development was divided in three stages i.e., immature (only spermatogonia, group I), early (spermatogonia and spermatocytes, groups II and III) and advanced (all germ cell stages including spermatozoa, groups IV and V) spermatogenesis. 11-Ketotestosterone blood levels were low, except for the last group. Antisera were raised against the complete catfish α,βGTH-II, as well as to the separate α- and β-subunits of catfish GTH-II. In the proximal pars distalis of immature fish, undifferentiated cells, somatotrops, putative thyrotrops (pTSH) and putative gonadotrops (pGTH) were found. In the two latter, secretory granules were labeled with anti-αGTH, but not with anti-βGTH-II. pTSH- and pGTH-cells were distinguished on the basis of the size of their secretory granules. During early spermatogenesis, two classes of putative gonadotrops could be distinguished. One type had the same immunocytochemical and ultrastructural characteristics as in immature fish; the secretory granules in the second cell type, which was more abundant, were also immunopositive for anti-βGTH-II. The mean volume of the secretory granules in these GTH-II cells was three times larger than that in the early appearing pGTH-cells. In addition, the later appearing GTH-II cells contained large inclusions, known as globules. These structures labeled with anti-αβGTH-II and with anti-βGTH-II, but not with anti-αGTH. It is assumed that the globules are involved in a differential storage and/or breakdown of the GTH-II subunits. During advanced spermatogenesis the two gonadotropic cell types could still be distinguished, but the early appearing pGTH-cell type was scarce. The present observations permit the conclusion that the early appearing cells may be GTH-I cells. However, definitive proof about their identity depends on the availability of antibodies or cDNA probes specific for GTH-I.     

Temperature and dietary factors affecting the nitrogen efflux rates of the African catfish, Clarias gariepinus (Burchell)

Total ammonia and total nitrogen efflux rates of the African catfish, Clarias gariepinus (Burchell), were measured under different temperature conditions (20, 25 and 30^°C) and feeding regimes (50% protein, 40% protein and non-fed for 10 days). Both ammonia and nitrogen efflux rates were directly related to temperature. At all experimental temperatures, the 50%-protein-fed groups excreted higher levels of ammonia nitrogen than both the 40% protein and non-fed groups. No correlation of relative ammonia efflux (% ammonia/nitrogen) with either temperature or diet was observed under these experimental conditions. Such alterations caused by temperature and diet may have significant implications for an aquaculture system as low levels of dissolved ammonia are synonymous with optimum water quality.     

Effect of light intensity on early ontogeny of African sharptooth catfish,Clarias gariepinus (Burchell)

Light intensity during the early life stages of fish can have profound effects on their survival, developmental rate, yolk utilization efficiency and body size. Here, these aspects were analysed during two separate experiments (with or without exogenous food) on two distinct progenies of African sharptooth catfish, at five different light intensities (<0.1, 70, 500, 2500 and 8000 Lx; 24L:0D, 27.2°C). The duration of the egg incubation period (from 1.01 to 1.25 days post fertilization, dPF) was inversely proportional to light intensity, as hatching took place at more precocious developmental stages with increasing light intensity, i.e. at significantly (P < 0.05) shorter body length and slightly more abundant remaining yolk at 8000 Lx in comparison to <0.1 Lx. At the start of exogenous feeding (4 dPF), most of these differences had vanished. During the period of mixed feeding (until the end of yolk absorption, 11 dPF), growth decreased significantly with increasing light intensity. Daily mortality rates after hatching varied very little between light intensities. Mortality during egg incubation increased significantly (P < 0.05) with increasing light intensity, whereas it varied very little between light intensities thereafter, with the best survival rates since fertilization until the end of yolk absorption obtained at intermediate light intensities (70–2500 Lx). These results could be useful for improving the performance of African sharptooth catfish hatcheries.     

Metabolic enzyme activities in larvae of the African catfish,Clarias gariepinus: changes in relation to age and nutrition

The influence of ontogeny and nutrition on metabolic enzyme activities in larvae of the African catfish, Clarias gariepinus, was studied. After start of exogenous feeding, the larvae were reared for 10 days under three different nutritional conditions: Artemia nauplii, a dry starter diet, and starvation. The live feed gave the best growth (96 mg within 10 days) whereas the dry diet resulted in low growth (33 mg). This growth difference was reflected in larval RNA and DNA concentrations, but not in the levels of soluble protein. Enzymes representing the following aspects of metabolism have been analysed: NADPH generation (G6PDH, ME), glycolysis (PFK, PK), gluconeogenesis (FDPase), amino acid catabolism (GOT, GPT) and oxidative catabolism (CS). All enzymes were present from the start of exogenous feeding onwards, but their maximum specific activities displayed different developmental patterns. In catfish larvae fed on Artemia, G6PDH and ME activities steadily increased with age and weight of the larvae. CS levels remained, after an immediate enhancement upon onset of exogenous feeding, on a rather stable plateau. The amino acid-degrading enzymes GOT and GPT showed maximum levels at days 3–5 of feeding or at a body weight of 10–20 mg, but decreased thereafter. Activities of PFK, PK and FDPase showed low initial levels, and increased significantly with age and size. Based on the ontogenetic patterns of metabolic enzymes, in C. gariepinus larvae an early and a late developmental phase can be distinguished. During the early phase, the glycolytic and gluconeogenetic capacities are low, whereas they are enforced during the later phase. The oxidative capacity is high both during the early and the late phase. The metabolic changes in catfish development coincide with other major ontogenetic events, e.g., alterations of muscle organization, gill morphology, respiration and stomach structure and function. Rearing catfish larvae on a dry diet instead of Artemia partly altered the developmental pattern described: The ontogenetic elevation of CS, PFK and FDPase was delayed and the early peak in GOT and GPT activities was not realized. Particularly during the early developmental phase, the enzyme behaviour of the larvae fed on dry food was similar to that of starved larvae.Abbreviations CS citrate synthase - FDPase fructose-1,6-diphosphatase - GOT glutamate oxaloacetate transaminase - GPT glutamate pyruvate transaminase - G6PDH glucose-6-phosphate dehydrogenase - ME malic enzyme - PFK phosphofructokinase - PK pyruvate kinase     

Yields of the African catfish,Clarias gariepinus (Burchell), from a low input,homestead, concrete pond

Fingerlings of the African catfish, Clarias gariepinus (Burchell), with a mean initial average weight of 96.2 g per fish were cultured in a dug-in homestead concrete pond, 30.24 m² in four cycles (1981–1984) for a mean average of 302 days. A mean average weight of 418.4 g per fish was obtained at harvest, resulting in an overall net fish yield of 40.3 kg. Homemade ration was used in the investigation and the feed conversion ratio was 2.6 to 1. There was a mean daily increase in individual fish weight of 1.06 g with overall fish survival of 86.4%.     

Characterization of the testicular semen of the African catfish, Clarias gariepinus (Burchell, 1822), and its short-term storage

Semen of the African catfish, Clarias gariepinus (Burchell, 1822), was investigated with respect to its cellular composition, sperm cell density, maturation grade, motility and fertility. Storage conditions were tested, whereby sperm viability was assessed by measurement of the motility after activation and by fertility tests. Testicular semen differed in its composition, i.e. the sperm density and numbers of spermatids, according to the maturity grade of the testis. Two semen types could be distinguished: semen type I was characterized by high sperm densities and low numbers of spermatids and semen type II had lower sperm densities and higher numbers of spermatids. Two semen types did not differ in motility and fertility (when adjusted for differences in sperm density). During storage, the sperm viability was influenced by the sodium concentration of the storage medium, temperature, membrane stabilizers as bovine serum albumen (BSA) or hen egg yolk, antibiotics and oxygen. Semen viability was maintained best when it was diluted at a ratio of 1:5 in storage solution (150 mmol L^?1 NaCl, 2.5 mmol L^?1 KCl, 1 mmol L^?1 CaCl₂, 1 mmol L^?1 MgSO₄, 20 mmol L^?1 Tris (pH 8.5) and 0.5% BSA or 0.5% hen egg yolk) and stored at 4 °C. Oxygen gassing and addition of antibiotics (1 mg mL^?1 gentamycine sulphate) to the storage solution affected the two semen types in different ways. Antibiotics had no effect on type I semen, but had a positive effect on type II semen. Oxygen gassing had a positive effect on type I semen but a negative effect on type II semen.