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European Journal of Biotechnology and Bioscience European Journal of Biotechnology and Bioscience Online ISSN: 2321-9122, Impact Factor: RJIF 5.44 www.biosciencejournals.com Volume 4; Issue 7; July 2016; Page No. 01-04 Thyroid and sex steroid hormone levels in the fresh water carps collected in wild from an aquatic body Raghavendra S Kulkarni, Pruthviraj CB Department of studies in Zoology, Gulbarga University, Kalaburgi, Karnataka, India Abstract The comparative studies on thyroid hormones triiodothyrinine (T3) ,thyroxin (T4),pituitary thyrotropin (TSH) and sex steroids (testosterone and estrogen) level has been estimated in four fresh water carp fishes, Labeo rohita, Catla catla, Cirrhana mrigala, Labeo fimbriatus collected from wild irrespective of sex before breeding period from a local aquatic body. The thyroxin (T4) is the predominant hormone than triiodothyrinine (T3) in all the carps studied with highest level in the carp fish, Cirrhana mrigala. The pituitary TSH level is uniformly same in all the carp fishes. Amongst the sex steroid hormones, testosterone is predominant hormone with higher levels in the carp fish, Catla catla than other three carp fishes. The estrogen level is less compared to testosterone in all the fishes with higher level in Catla catla compared to other carp fishes. Although all the carp fishes exhibited normal levels of the hormones studied, the predominant hormones are thyroxin (T4) and testosterone indicating that all the fishes belong to males. Keywords: Haematology, Indian carps, Labeo rohita, Catla catla, Cirrhana mrigala and Labeo fimbriatus 1. Introduction Hormones catalyze and control diverse metabolic processes. Hormones are not only catalysts for all metabolic process, they are living proteins that direct the life force into over basic biochemical and metabolic process. The hormones help transform store energy and activate other physiological process including their participation in reproductive activities. The regulatory actions of hormones are related to homeokinetic reactions of metabolism, salt and water balance etc. The study pertaining to role of different kinds of hormones in various physiological activities in fish has been reported. The study related to changes in the hormone level at different phases of reproductive cycle have been reviewed (Hontela and Stacey,1990., Degani Boker, 1992), very few Indian fishes have been worked on such aspects (Singh and Singh, 1987) Although the changing titers of gonadal hormones during different period of the reproductive cycle of the fish has been studied and reviewed by several investigators, (vide Malhotra et al, 1989 ), the studies related to the hormone levels of different species of fish or fishes belonging to the same family group are not available. Hence, in the present investigation locally available four freshwater carp fishes, Labeo rohita, Catla catla, Cirrhana mrigala, Labeo fimbriatus collected from wild irrespective of sex before breeding period from a local aquatic body ware used for the determination of hormonal level on a comparative basis and in relation to the particular aquatic body. 2. Materials and Methods The freshwater fishes were collected during the prebreeding (preparatory -March) phase. The fish were collected with the help of local fisherman from Khaja kotnoor reservoir near Gulbarga. The blood was collected from live fishes separately. The serum content of hormone such as thyrotropin (TSH), T3, T4, estrogen and testosterone were estimated by applying CLIA (Chemiluminescence Immunoassays) method during pre-breeding phase. The hormone assays were carried out at M/s Diagnostic Hormone Laboratory, Gulbarga. For the Chemiluminescence immunoassay of pituitary thyrotropin hormone (TSH,) thyroid hormones (T3 and T4) and steroid hormones (Estrogen, testosterone) the fish taken to the laboratory and each fish were sacrificed by cutting at the caudal region. The flowing blood was collected in to the test tube and is used for further processing. The amount of blood sample required for the estimation of each hormone is as follows: Pituitary hormones TSH Thyroid hormones Triidothyronine Thyroxine Sex steroid hormones Estrogen Testosterone 100 L 100 L 25 L 100 L 50 L 3. Observation The comparative serum concentration of hormones such as pituitary thyrotropin, thyroid and sex steroids in the carp fishes collected in wild irrespective of sex has been estimated during pre-breeding phase. The pituitary hormones such as thyrotropin, thyroid hormones such as triiodothyrinine (T3) and thyroxin (T4), gonadal hormones such as estrogen, and testosterone has been estimated separately. The comparative serum concentrations of these hormones are presented in the Tables-1, Figs.1 & 2. Thyroid hormone (T3 and T4): Triiodothyrinine (T3) and tetraiodothyronin (T4 – Thyroxin) level presented in the Table-1 and Fig.-1 & 2 for all the four carp fishes indicate that T4 concentration is higher than T3 in all the fish during prebreeding phase. The increased level of the hormone during prebreeding phase indicates energy requirement during the phase. The fish, Cirrhana mrigala has higher level of the hormone thyroxin (T4) compared to other carp fishes. The thyroxin (T4) may be the predominant 1 European Journal of Biotechnology and Bioscience hormone in all the fishes. The pituitary TSH level is uniformly same in all the fishes. Sex steroid hormones: Testosterone: Table-1and Fig.-2 shows serum concentration (mg/ml) for four fishes. In all the four carp fishes testosterone level is higher compared to estrogen hormone. The higher level of the hormone testosterone in all the fish during pre-breeding phase suggests its role in spermatogenesis. Estrogen: Table-1 and Fig.-2 shows serum level of estrogen concentration (pg/ml) values for all four fishes. The estrogen level is less compared to testosterone in all the carp fishes with higher level in catla catla compared to other fishes. Although all the carp fishes exhibited normal levels of the hormones studied, the predominant hormones are thyroxin (T4) and testosterone indicating that all the carp fishes caught may be belonging to males. The study also suggests that the fish, Cirrana mrigala is very active fish and Catla catla are more reproductive than other carp fishes. Table 1: Showing blood thyroid and gonadal hormones in four Indian carp fishes collected from a aquatic body Fish Species T3 T4 TSH Testosterone 1.17 ± 0.163 90.45±0.008 0.20±0.012 147.00±1.673 1 Labeo rohita (SE=0.007 ) (SE=0.003) (SE=0.004) (SE=0.683) 0.98 ± 0.008 79.45±0.014 0.01±0.000 335.83±0.752 2 Catla catla (SE=0.003) (SE=0.006) (SE=0.000) (SE=0.307) 1.16 ± 0.008 97.42±0.008 0.08±0.008 201.00±1.264 3 Cirrhana mrigala ( SE=0.003) (SE=0.003) (SE=0.003) (SE=0.516) 1.01 ± 0.009 85.96±0.005 0.28±0.008 196.83±1.169 4 Labeo fimbriatus (SE=0.004) (SE=0.002) (SE=0.003) (SE=0.477) Each value is expressed as mean ± SD, N = 6. All values are significant P = < 0.01 Estrogen 95.50±1.378 (SE=0.562) 121.16±1.169 (SE=0.477) 96.66±1.632 (SE=0.666) 101.16±0.983 (SE=0.401) Fig 1: Showing blood level of thyroid triiodothyrinine (T3) and pituitary thyrotropin (TSH) hormones in four Indian carp fishes collected from a aquatic body Fig 2: Showing blood level of thyroid- thyroxin (T4) and gonadal hormones (Testosterone and Estrogen) in four Indian carp fishes collected from a aquatic body 2 European Journal of Biotechnology and Bioscience 4. Discussion The plasma levels of various hormones produced from pituitary and other endocrine glands in relation to seasonal gonadal development and maturation have been reported in several species of fish which has proved to be valuable tool in determining the role of hormones in growth in the fish. These studies also provide information in understanding of endocrine control of physiology of fishes. Matty, (1985) has discussed the role of thyroid in teleost reproduction. Seasonal thyroid hormone levels have been measured in salmonids. In the trout Salmo gairdneri. There appears to be two peaks in plasma concentration (Osborn et al., 1978), while in Lake Ontario cohosalmon, there is but one peak of serum T4 and T3. It has been further suggested that two thyroid cycles occur in teleost fish, one concerned with temperature compensating mechanism and the other with reproduction. The thyroid hormones (T3 and T4) measured in the present study indicates that T4 level is higher than T3 level in all carp fishes. The fish Cirrhana Mrigala has still higher level compared to other carp fishes probably indicates that the fish is on constant movement, needs higher metabolic rate which may be the reason why the thyroid hormone found to have higher level compared to the other three fishes. The increase in the level of the hormone during prebreeding phase may be related to still higher metabolic rate during high temperature period in this are (Gulbarga where environmental temperature goes upto 46 degree or more) and it may be possible that the fishes regulate temperature compensatory mechanism as reported by Matty (1985), or it may be also for approaching active reproductive phase. A number of workers have reported biosynthesis of steroids from the gonads of variety of fishes and generally steroid hormones such as 17-estradiol, testosterone estimated in the present study in the carp fishes have been also identified in number of teleosts (Vide, Fostier et al., 1983) during different phases of reproductive cycle. The ovarian steroid hormones in females includes 17-estradiol, testosterone and progesterone along with other androgens and estrogens (Colombo et al., 1982; Fostier et al., 1983). Testicular steroid hormones in males include testosterone, estradiol 17 and progesterone along with other androgen and estrogens (Vide Fostier, et al., 1983). The steroid hormones such as estrogen and testosterone have been also detected and quantitatively determined by RIA using coat a method in carp fishes in the present investigation during pre-breeding phase. Correlation between seasonal changes in plasma levels of gonadal steroids and gonad condition have been well documented in a number of freshwater fish species (Van der Merwe et al., 1987). Steroid hormone levels particularly estradiol-17 and testosterone levels were low in postspawning whle they gradually increased during preparatory and prespawning phases in both the sexes. In the present study although testosterone level is higher than estrogen, they are at elevating since all the fishes are at prebreeding phase. Similar study has been also reported on the seasonal variation of steroid hormone level in an intertidal resting fish, plain fish midshipman (a deep water teleost fish) the plasma levels of testosterone and estradiol-17 were determined and found very low throughout the year and peaked during ovaries underwent seasonal recrudescence (Sisneros et al., 2004). Estrogen synthesis, mainly 17-estradiol and / or estrone has been found in most teleosts examined (Fostier et al., 1983). The estradiol-17 is secreted from the gonads in both male and female fish. In general estradiol is responsible for stimulating vitellogenesis and is also secreted by male gonads. Estradiol has been reported to stimulate vitellogenesis in teleosts (Campbell and Idler, 1976; De Valming et al., 1980; Smith and Haley, 1988). There is decrease in the level of estradiol during spawning phase compared to preparatory and prespawning phase it may possibly indicate that a rapid utilization of the hormone in stimulating vitellogenesis or completion of vitellogenesis as the fish undergoes spawning activity. In the fish Oreochrosmis mossambicus (Cornish, 1998) reported that during resting season there are generally low levels of steroid hormones and during ovarian recrudescence, there is an increase in trophic and steroid hormones and suggests that this relates to vitellogenesis. The plasma testosterone levels in carp fishes in the present study found to be higher during preebreeding phase indicates its role in gonadal growth particularly for spermatogenesis The testosterone levels are higher in Cirrhana mrigala than other fishes corresponds to its requirement for spermatogenesis . The general increase in testosterone during prebreeding phase in this study indicate the commencement of spermatogenesis within the testis, such increase level of testosterone in both male and female fish has been reported in the fish, O. mossambicus (Cornish, 1998) and suggested its need for spermatogenesis in males and growth of gonads in females. Earlier studies also show that the androgen production from the ovaries has been reported in fish species in vitro studies (Fostier et al., 1983). In the grey mullet, Mugil cephalus, the ovarian production of the ketotestosterone increases with the development of vitellogenesis and then decreases after spawning (Azoury and Eckskin, 1980). 5. Conclusions The blood hormone level studied in four species of fishes such as Labeo rohita, Cutla cutla, Cirrhana mrigala and Labeo fimbriatus indicates that the values are more or less normal in comparison to other fishes reported. Thus indicating all the four types of fishes is healthy and regulating normal endocrine activity. There is variation of hormone values between the fish species studied and this could be the species specificity that should be considered in accepting study specimens and evaluating results. 6. Acknowledgement The author R.S. Kulkarni is grateful to University Grants Commission, New Delhi for selecting Emeritus fellowship (2014-15 batch) No.F-6-6/2014-15/EMERITUS-2014-15GEN-3514 (SA-II) Dt.30-04-2015.we are also thankful to The Chairman of the Department and to Gulbarga University for the facilities to carry out this work and also to M/S. Pooja Diagnostic Laboratory, Gulbarga. 7. References 1. Azoury R, Eckstein B. Steroid production in the ovary of the grey Mullet Mugil cephalus during stages of egg ripening. Gen. Comp. Endocrinol., 1980; 42:244-250. 2. Campbell CM, Idler DR. 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