Thyroid hormone modulation of ovarian recrudescence of air-breathing catfish Clarias gariepinus

In the present study, thiourea-induced thyroid hormone depletion and thyroxine (T₄) ‘overdose’ were used as a strategy to understand the influence of thyroid hormones on ovarian recrudescence of juvenile (3-months-old), immature (8-months-old) and adult (1-year-old) air-breathing catfish, Clarias gariepinus. Thiourea-induced thyroid hormone depletion in juvenile catfish impaired ovarian development, but no significant effect was observed in immature catfish and during late stage of ovarian recrudescence of mature catfish. T₄ treatment in females undergoing late stages of ovarian recrudescence induced rapid oocyte growth by promoting its early entry into maturational phase as evident from the presence of more number of vitellogenic and post-vitellogenic follicles, decrease in aromatse immunoreactivity and reduced estradiol–17β levels. Hence, thyroid hormones have an important role to play during early stages of ovarian development and vitellogenesis of catfish and also indicating that thyroid has a stage dependent effect on ovary.     

Thyroxine-induced alterations in the testis and seminal vesicles of air-breathing catfish, Clarias gariepinus

Effect of experimentally induced thyroxine overdose on the testis and seminal vesicles was studied in the air-breathing catfish, Clarias gariepinus during the preparatory and the pre-spawning phase. The present study revealed a marked reduction in testosterone level in serum, testis and seminal vesicles (SV). Histological examination showed a considerable reduction in the number of spermatozoa/spermatids in the seminiferous tubular lumen as well as depletion of fluid in the loculi of SV. SDS-PAGE analysis of SV fluid proteins demonstrated a significant decrease in the level of a ~27 kDa protein in thyroxine treated fishes. Evidences are presented here to indicate that thyroid hormone plays a role in regulating testis and SV function in catfish. T.N. Jacob and J.P. Pandey contributed equally     

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.     

Vitamin C deficiency in the catfish Clarias gariepinus

Regardless of the absence of spinal deformities, vitamin C deprivation in adult-sized Clarias gariepinus challenged the physical integrity and survival of fish, inducing changes that greatly resembled those of fingerlings of other species. Vitamin C was presumed to be a dietary facilitator, which when deprived in the diet of C. gariepinus , would obstruct intestinal absorption of iron and/or impair its release from the reticulo-endothelial stores. This impaired erythrocyte synthesis led into anaemia (as indicated by lowered levels of red blood cells, haemoglobin and haematocrit). Leukopenia noted in fish on a scorbutic (lacking vitamin C) diet suggested functional depression of phagocytic engulfment with increased susceptibility to the deteriorating action of pathogenic agents. Changes in histology included hypertrophy, oedematous separation and telangiectasia of gill lamellae. Shrunken glomeruli, sloughed Bowman's capsules and tubular dilation and distortion were also shown. Shrunken hepatocytes and the occurrence of yellowish ceroid pigments were the concrete histopathological signs in liver. Vitamin C is thus suggested as a potent antioxidant that offers protection against oxidative damage to various fish tissues.     

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.     

Effect of microalga-based diet on oxidative stress enzymes of African catfish,Clarias gariepinus

Here an indigenously isolated microalgal strain Ascochloris spp. cultivated in synthetic medium was evaluated as an aquaculture feed supplement. The daily dietary supplement includes microalgal feed (AF) and commercial diet feed (CF) (as control), respectively. These diets were fed separately to the juvenile Clarias gariepinus fishes (n = 4) under controlled conditions for an experimental period of 100 days. The protein, glycogen and lipid contents in the muscle extracts were found to be marginally higher in fishes that were fed with CF than AF diet. Similarly, CF fishes showed significantly higher glutathione-s-transferase, catalase, superoxide dismutase, and lipid peroxidase activities, except glutathione content. Zero mortality of the fishes with no significant difference in the overall body mass with the two dietary supplements strongly suggests that algal biomass could supplement the requisite nutrients for their metabolic activities. This preliminary investigation helps in exploring algal biomass as a potential alternative feed additive in the aquaculture industry.     

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.     

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.     

Effect of extenders on sperm cryopreservation of African catfish, Clarias gariepinus (Burchell)

The effect of extenders was studied on the cryopreservation of sperm from African catfish, Clarias gariepinus (Burchell). The following six basic extenders were tested: fructose, glucose, sucrose, NaCl, KCl solutions and the artificial seminal plasma of the African catfish. Each of these extenders was tested both with and without buffer systems (i.e. NaHCO₃-CO₂ and Tris-HCl) by using 10% dimethyl-sulphoxide (DMSO) as a cryoprotectant. The two-step freezing was carried out in a programmable freezer by using the following freezing rates: (1) 4 °C min^–1 between 3 and –4 °C; (2) and 11 °C min^–1 between –4 and –80 °C. The best post-thaw motility (25%) was achieved with 333 mmol L^–1 fructose solution and NaHCO₃ buffer. The fertilization experiments were carried out with unbuffered fructose and glucose extenders using various amounts of sperm and two fertilization methods: (1) dry and (2) wet. The best fertilization rates were achieved with 75 μL of sperm and wet fertilization with glucose extender, or 100 μL of sperm and dry fertilization in case of fructose – both methods fertilized 96% of all eggs.     

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     

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.     

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.     

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.     

絮团浓度对革胡子鲇零换水养殖效果的影响

为研究絮团浓度对革胡子鲇零换水养殖效果的影响,在不额外添加有机碳源（只利用饲料中的碳）的革胡子鲇（）养殖系统中,设置了平均絮团质量浓度为561.18 mg/L和780.41 mg/L两个处理组,比较了两实验组的水质、菌群结构、鱼生长及氮利用效率。结果表明,两种浓度絮团条件下,总氨氮（total ammonia nitrogen,TAN）和亚硝酸氮（NO₂^--N）能分别维持1.84 mg/L和1.79 mg/L以下。两处理组间pH、溶解氧（dissolved oxygen,DO）、TAN、NO₂^--N、氮素利用效率及主要生长指标无显著差异（-N）浓度（822.0 mg/L）明显高于低浓度絮团组（623.33 mg/L）。高通量测序分析菌群结构结果表明,两组间门水平的菌群组成种类及优势度无显著性差异（<0.05）。两处理组中的革胡子鲇存活率分别达到（91.11±1.53）%和（94.44±2.08）%,饲料系数为（1.41±0.18）和（1.27±0.26）,特殊生长率为（2.13±0.04）%/d和（2.19±0.08）%/d,均无显著差异（>0.05）。两实验组饲料氮的利用率分别达到了72.17%和71.34%。综合以上结果认为,仅利用饲料中的碳既能维持革胡子鲇的零换水养殖且能取得较高的氮素利用效率,两种絮团浓度对革胡子鲇的生长无显著影响,高浓度絮团组中的硝化作用更明显。     

Preliminary studies on the renaturation of denatured catfish (Clarias gariepinus) glutathione transferase

Purified juvenile catfish (Clarias gariepinus) glutathione transferase (cgGST) was denatured in vitro and renatured in the absence and presence of different concentrations of endogenous or xenobiotic model substrates. Protein transitions during unfolding and refolding were monitored by activity measurement as well as changes in protein conformation using UV difference spectra at 230 nm. Gdn-HCl at 0.22 M caused 50 % inactivation of the enzyme and at 1.1 M, the enzyme was completely unfolded. Refolding of cgGST main isozyme was not completely reversible at higher concentrations of Gdn-HCl and is dependent on protein concentration. An enzyme concentration of 30 μg/ml yielded 40 % percentage residual activity in the presence of glutathione (GSH), regardless of the concentration that was present as opposed to 30 % obtained in its absence. The xenobiotic model substrate, lindane, appears to have no effect on the refolding of the enzyme. In summary, our results show that GSH assists in the refolding of cgGST in a concentration-independent manner and may be involved in the same function in vivo whereas the xenobiotic model substrate does not.     

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.     

Role of cathepsins B and D in proteolysis of yolk in the catfish Clarias gariepinus

Fish Physiology and Biochemistry - Yolk processing pathways vary in the oocytes of benthophil and pelagophil teleosts. The present study investigated the yolk processing pattern in the oocytes of...     

Steroids and steroid glucuronides in the ovarian fluid of the African catfish,Clarias gariepinus,between ovulation and oviposition

As part of a series of experiments concerning a possible pheromonal function of steroids and steroid glucuronides excreted by the sex organs of the African catfish,Clarias gariepinus, qualitative and quantitative studies, using GCMS, were carried out to examine the presence of the steroids, that can be synthesized by the ovary during oocyte maturation and ovulation, and of the corresponding steroid glucuronides, in the fluid surrounding the eggs in the ovarian cavity shortly after ovulation.Full mass spectra were obtained of 5-pregnane-3,17-diol-20-one, 5-pregnane-3,17,20-triol, 5-pregnane-3,6,17-triol-20-one, 5-pregnane-3,6,17,20-tetrol, 5-androstane-3,17-diol and 5-androstane-3,17-diol-11-one. After selected ion monitoring the following steroids could be detected by the presence of at least two characteristic ions at the expected retention time: 5-pregnane-3, 17,20-triol, etiocholanolone, 5-dihydrotestosterone, 5-androstane-3,11-diol-17-one, testosterone and estradiol. After treatment with -glucuronidase the following steroids could be determined in a similar way: 5-pregnane-3,17-diol-20-one, 5-pregnane-3,17,20-triol, 5-pregnane-3,17,20-triol, 5-pregnane-3,6,17-triol-20-one, 5-pregnane,3,6,17,20-tetrol, 5-androstane-3,17-diol, etiocholanolone, 5-dihydrotestosterone, testosterone and estradiol.The free steroids 5-pregnane-3,6,17,20-tetrol and 5-pregnane-3,6,17-triol-20-one and the steroid glucuronides of testosterone, 5-dihydrotestosterone and estradiol appeared to be the most abundant of these compounds. The results indicate that very polar steroids and steroid glucuronides, synthesized in the ovary, can be excreted via the ovarian fluid shortly before and during oviposition, and possibly function as sex attractants, inducing reproductive behaviour in male conspecifics.