Bechan Lal

Changes in tissue lipid levels in the freshwater catfishClarias batrachus associated with the reproductive cycle

The levels of free fatty acid (FFA), monoglyceride (MG), diglyceride (DG), triglyceride (TG), phospholipid (PL), free cholesterol (CF) and esterified cholesterol (CE) were measured in the liver, plasma, ovary, abdominal fat and muscle during different phases of the annual reproductive cycle in femaleClarias batrachus. During the preparatory phase, hepatic lipogenic activity predominated over mobilization and consumption. In the prespawning season, an increased hepatic lipogenic activity was maintained, but lipids were transferred from the liver to the ovary. In the spawning phase, the diminished food intake, and enhanced caloric demand for spawning behaviour and activity limited hepatic lipogenic activity, and TG lypolysis was increased as was the production of more FFA. Maximum accumulation of vitellogenin, as reflected by maximum rise in ovarian PL titre was characteristic of this phase. Marked reductions in ovarian lipid occurred during the postspawning phase. In the resting phase, there was a recovery of lipogenic activity, but PL synthesis was still inhibited. In contrast to other investigated teleosts, there were extremely high level of FFA in liver, plasma, ovary and muscle throughout the annual reproductive cycle inC. batrachus. FFA appears to be the main lipid metabolite which had a very high turnover. As evidenced by the high TG content, abdominal fat seems to be the main fat depot, not the liver and muscle.

Effect of endosulfan (thiodan) on vitellogenesis and its modulation by different hormones in the vitellogenic catfish Clarias batrachus

This programme was planned in order to study the reproductive dysfunction caused by endosulfan in the fish Clarias batrachus with special reference to vitellogenesis. Vitellogenin was measured by radioimmunoassay. Endosulfan caused a drastic reduction in the plasma vitellogenin which was partially reversed by estradiol-17 beta (E2) in intact and ovariectomized fish. Hormones in combination were more effective than any single hormone in overcoming the impact of endosulfan. Triiodothyronine (T3), ovine luteinizing hormone releasing hormone (oLHRH) were effective in minimizing the effect of endosulfan whereas cortisol (F) and ovine somatotropic hormone (oStH) did not reduce the effect of endosulfan. Amongst the combined hormone treatments, E2 + oLHRH and E2 + T3 were the most potent in overcoming the effect of endosulfan on vitellogenesis.

Pesticides induced changes in circulating thyroid hormones in the freshwater catfish Clarias batrachus

1. Effects of endosulfan, malathion and carbaryl on circulating T4, T3 and T3/T4 ratio in the freshwater catfish, C. batrachus have been studied during reproductively active phases, i.e. vitellogenic and post-vitellogenic phases of its annual reproductive cycle. 2. Endosulfan provoked a significant increase in T4 and a decrease in T3 as well as the T3/T4 ratio. 3. Malathion decreased T3 and T3/T4 ratio without altering the level of T4 in circulation. 4. Carbaryl caused a decline in the level of T4 and an elevation in T3 and T3/T4 ratio.

Effect of endosulfan on thyroid physiology in the freshwater catfish, Clarias batrachus

Effects of endosulfan on thyroid physiology have been studied in the female freshwater catfish. Clarias batrachus during the pre-spawning and spawning phases of its annual reproductive cycle. Effects of endosulfan varied with the length (96 h and 16 days) of exposure, reproductive status of the fish and organ. The 96-h endosulfan exposure significantly increased the level of thyroxine (T4) in serum and pharyngeal thyroid follicles concurrent with induction of peroxidase activity. However, triiodothyronine (T3) level and T3/T4 ratio decreased in serum and pharyngeal thyroid gland. No change was noticed in any of these parameters in the anterior kidney but in the posterior kidney endosulfan reduced T3 and T3/T4 ratio without affecting T4 levels and peroxidase activity. Sixteen days of endosulfan treatment also had a similar impact, except that it did not influence the studied parameters in pharyngeal thyroid.

Carbaryl-induced thyroid dysfunction in the freshwater catfish Clarias batrachus

Freshwater female catfish, Clarias batrachus, were exposed to sublethal concentrations of carbaryl (12 mg/liter for 96 hr and 5 mg/liter for 16 days) during the prespawning and spawning phases of their annual reproductive cycle. The effects on the levels of thyroxine (T4) and triiodothyronine (T3), the T3/T4 ratio, and the extrathyroidal conversion of T4 to T3 were studied in serum, pharyngeal thyroid, and anterior and posterior kidney. Carbaryl exposure for 96 hr suppressed T4 but elevated T3 and the T3/T4 ratio in serum during both phases of experimentation. However, 16 days exposure caused a decline in T4 as well as in T3 and in the T3/T4 ratio. In the pharyngeal thyroid, carbaryl increased T4 as well as peroxidase activity and decreased T3 and the T3/T4 ratio. Its treatment for 96 hr did not elicit any change in the parameters studied in the anterior kidney, whereas in the posterior kidney it suppressed the levels of T4 and T3, peroxidase activity, and the T3/T4 ratio. However, carbaryl exposure for 16 days provoked increases in T4 and peroxidase activity but decreases in T3 and the T3/T4 ratio in anterior kidney. In posterior kidney, it did not change T4 and peroxidase activity, but decreased T3 and the T3/T4 ratio. It also stimulated extrathyroidal conversion of T4 to T3.

Malathion exposure induces the endocrine disruption and growth retardation in the catfish, Clarias batrachus (Linn.)

Many hormones are known for their role in the regulation of metabolic activities and somatic growth in fishes. The present study deals with the effects of malathion (an organophosphorous pesticide) on the levels of metabolic hormones that are responsible for promotion of somatic and ovarian growth of the freshwater catfish, Clarias batrachus. Malathion treatment for thirty days drastically reduced the food intake and body weight of fish. These fish also exhibited a great avoidance to food. Exposure of catfish to malathion reduced the levels of thyroxine (T(4)), triiodothyronine (T(3)), growth hormone (GH), insulin like growth factor-I (IGF-I), testosterone (T) and estradiol-17β (E(2)) in a dose dependent manner during all the studied reproductive phases, in general, except that malathion increased the level of GH during the quiescence phase. Significant reduction in muscle and hepatic protein content also occurred in the malathion-treated fish. Malathion exposure induced lipolysis too in the liver and muscle. The results thus support that malathion treatment disrupts the endocrine functions and the olfactory sensation responsible for food intake and gustatory feeding behavior, which ultimately leads to retardation of fish growth.

Seasonality in expression and distribution of nitric oxide synthase isoforms in the testis of the catfish, Clarias batrachus: role of nitric oxide in testosterone production.

Nitric oxide (NO) is a well-recognized versatile signaling molecule. It is produced by catalytic action of nitric oxide synthase (NOS) on L-arginine in a variety of animal tissues. Existence of different isoforms of NOS has been shown in mammalian testis, but report on their presence in the testis of ectothermic vertebrates is non-existent. This study demonstrates the differential expressions of two isoforms of nitric oxide synthase (neuronal-nNOS and inducible-iNOS) like molecules in different cell types in the testis of seasonally breeding catfish, Clarias batrachus through immunohistochemistry. Positive immunoprecipitation of nNOS and iNOS like molecules were detected in germ cells as well as interstitial cells only in the recrudescing and fully mature fish. The immunoreactions differed in intensity and varied with changing reproductive status. Treatment of adult male fish with NO donor, sodium nitroprusside, and a NOS inhibitor, N-nitro-L-arginine methyl ester (L-NAME) increased and decreased the total nitrate and nitrite concentration in the testis, respectively. Sodium nitroprusside and L-NAME also induced simultaneous decline and rise in the testicular testosterone level, respectively. These findings, thus, suggest that NOS isoforms are expressed variedly in different cell types in the testis of reproductively active fish. This investigation also suggests that NO inhibits testosterone production in the testis.

The effect of malathion and γ-BHC on the lipid metabolism in relation to reproduction in the tropical teleost, Clarias batrachus

In the present study, female Clarias batrachus were exposed to two sublethal concentrations of gamma-BHC (2 and 8 microg litre(-1)) and malathion (1 and 4 microl litre(-1)) for 4 weeks during different phases of their reproductive cycle. Impact of these pesticides on free fatty acids, monoglycerides, diglycerides, triglycerides, phospholipids, free and esterified cholesterol in the liver, plasma, ovary and muscle was assessed. During the pre-vitellogenic and regressed phases, they suppressed the levels of fatty acids and glycerides in the liver, but had no effect on their levels in the plasma, ovary and muscle. However, in the vitellogenic phase, fatty acids in the liver and plasma were increased, but were decreased in the ovary and muscle. Glycerides were also decreased in the studied tissues. In the post-vitellogenic phase, these pesticides increased the levels of fatty acids and glycerides in the liver and ovary, but decreased their levels in the plasma. Both pesticides decreased hepatic phospholipids in the pre-vitellogenic phase, while in the vitellogenic phase only gamma-BHC reduced its levels in the liver. These pesticides also restricted their mobilisation to the ovary. Cholesterol biosynthesis seemed unaffected, but the hydrolysis of esterified cholesterol to free cholesterol was adversely affected during the period of estrogen biosynthesis.

Impact of pesticides on lipid metabolism in the freshwater catfish, Clarias batrachus, during the vitellogenic phase of its annual reproductive cycle

Specimens of either sex of the freshwater catfish Clarias batrachus were exposed to safe and sublethal concentrations of an organochlorine, gamma-BHC (2 and 8 ppm), and an organophosphorus compound, malathion (1 and 4 ppm), for 4 weeks during the vitellogenic phase of their annual reproductive cycle. The effects on total lipid and its various fractions, viz., free fatty-acids, monoglycerides, diglycerides, triglycerides, phospholipids, free cholesterol, and esterified cholesterol, were studied in the liver, plasma, gonads, and muscle. Except for elevated liver lipid in the male in response to malathion, no significant change in total lipid could be observed following pesticide exposure. However, various lipid fractions responded differently to two concentrations of the pesticides. Both pesticides affected the metabolism of nonpolar and less polar lipids alike. Malathion inhibited only mobilization of hepatic phospholipid to gonads but not its hepatic biosynthesis, whereas gamma-BHC reduced its synthesis in the liver as well. These pesticides seemed to restrict the conversion of esterified cholesterol into free cholesterol without affecting the biosynthesis of cholesterol as such. In both sexes, esterification of free fatty acids to acyl glycerides and their mobilization from liver to gonads seemed to be restricted as a result of pesticides action.

Nitric oxide: An autocrine regulator of Leydig cell steroidogenesis in the Asian catfish, Clarias batrachus.

Nitric oxide has been recognized as an important inter- and intra-cellular modulator of testicular steroidogenesis in higher vertebrates with conflicting results. Moreover, its role in regulation of testicular steroidogenesis in ectothermic vertebrates is non-existent. The present study was, therefore, undertaken to examine whether Leydig cells of a freshwater catfish, Clarias batrachus produce nitric oxide (NO), if so, does it regulate its steroidogenic activity? The purified Leydig cells were stained histochemically for NADPH-diaphorase (NADPH-d) activity, and immunocytochemically for neuronal and inducible nitric oxide synthase (nNOS and iNOS) like molecules. Leydig cells were also incubated with NOS inhibitor, N-nitro-l-arginine methyl ester (l-NAME), and NO donor, sodium nitroprusside (SNP). NO and testosterone released in incubation medium were analyzed. A distinct positive NAPDH-d staining was observed in Leydig cells. These cells also exhibited immunoprecipitation of variable intensity with nNOS and iNOS antibodies. Further, l-NAME treatment caused significant suppression in NO production and elevation in testosterone secretion by Leydig cells. On the contrary, exposure of Leydig cells to SNP resulted in increased NO production with concomitant decline in testosterone level. Thus, the present study reports NO production by Leydig cells in fish for the first time, which appears to inhibit its own androgen production.