The relationship between water quality parameters and response to density stress in Pacu fish (Piaractus brachypomus)

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International Research Journal of Applied and Basic Sciences 2013 Available online at www.irjabs.

1 International Research Journal of Applied and Basic Sciences 2013 Available online at ISSN X / Vol, 4 (6): Science Explorer Publications The relationship between water quality parameters and response to density stress in Pacu fish (Piaractus brachypomus) Majid Naserizadeh 1, Mohammad Ali Nematollahi *1, Seyed Vali Hosseini 1 1. Department of Fisheries, Faculty of Natural Resources, University of Tehran, Karaj, Iran. * Corresponding Author malahi@ut.ac.ir ABSTRACT:Backgrounds: Pacu, Piaractus brachypomus is an ornamental imported fish that is reared as an edible fish in some countries. There have been very few studies as food fish in Iran. Objectives: This study aimed to evaluate the relationships between water quality parameters and stress response indices in different densities in a relatively recirculation system. Methods: An experiment carried out in 3 recirculation treatments with 3 replications in 250-liter tanks with a filled volume of 200 litres water. An extra 100-liter tank prepared as biological filter. Tanks were stocked in 3 densities 50, 75 and 100 fingerling.m -3 with an average weight as 3.52±0.11 for duration of 3 months. Water replacement was 10% per day. Water quality parameters of total ammonia, nitrite and nitrate determined every 15 days and dissolved oxygen, alkalinity and electrical conductivity every day. Blood collected to determine plasma cortisol and glucose to evaluate the effect of water quality on stress response in different densities. Results: The results showed that in densities 50 and 75 fish/m 3, there was no significant effect on stress response indices of cortisol and glucose (P>0.05). But, alkalinity decreased nitrite and nitrate parameters increased in 100 fish/m 3 (P<0.05). Likewise, in this density stress response indices of cortisol and glucose significantly increased in third month of rearing. Conclusions: It suggests that density of 100 fish/ m 3 can be a chronic stressor for fish which raise cortisol response. Key Words: Pacu, Recirculation system, Water quality, Cortisol, Glucose. INTRODUCTION Nowadays, uses of recirculation systems due to their advantages to other rearing systems are increasing. Among them are increasing of production efficiency and reduction of rearing time in recirculation systems a very little amount of the water after circulating in system goes out of the system and the rest of that after refining come back to the system again (Timmons et al., 2002). But sometimes, low water exchange in such systems causes some problems such as depletion of dissolve oxygen and accumulation of metabolites producing by fish (Sanni and Forsberg, 1996). To avoid these problems and in order to keep water quality should be prevented than accumulation of the suspended organic matter and also ammonia, alkalinity, temperature and dissolved oxygen should be maintained at an acceptable level appropriate with cultural species because lack of water appropriate quality is one of most common and important reasons for fish stress that it can easily be avoided by taking measures (Schreck and Li, 1991). If reduction of physical and chemical quality parameters of water is more than those fish which can tolerate it, they are forced to spend energy to overcome this situation because stress is a process that needs energy and animals to overcome it use of their energy storage (Wedemeyer, 1997). Fish through releasing glucose during stress period improve the amount of energy demand for more efficiency of brain, gills and other organs until they be able to deal with difficult situations in a better condition and this causes further damage to the physiological system (Barton, 2002).Therefore, if stressors are in the environment for a long time, they will cause reduction in growth, incidence of various disease, failure in reproduction and etc. (Barton, 1997). Various species of fishes can have different responses to different stressors (Pottinger and Pickering, 1997). Methods of fish culture are also effective in their responses to releasing of glucose (Vijayan and Moon, 1992

2 and 1994). Thus, study of fish stress factors in various species and different conditions of fish culture need to have separate experiments. In the present study the relationship between water quality parameters and stress factors in the different densities of P. brachypomus in recirculation systems was examined. This species has a good potential for fish farming in recirculation systems. This study can help us in introducing and improving the condition of fish farming in recirculation systems. MATERIAL AND METHODS This experiment was done in the research section of Tange Tizab rainbow trout aquaculture center in Sepidan city (Fars, Iran) on Pacu fingerlings with primary weight of 3.52 ± 0.11 g. Experimental condition In the experiment three treatments of density was used. First treatment consisted of 10 fish, second treatment consisted of 15 fish and third treatment consisted of 20 fish (50, 75 and 100 fish per m 3 respectively). Treatments were done in triplicates. The water that existed in the each treatment reservoirs associated with each other recirculation and 10% of their water was changed daily. Water temperature in reservoirs was kept constant at 27 C by a heater that warmed the air in the experimental place and photoperiod was 16 L: 8 D. After an adaptation period for a week, the fish reared in such conditions for three months and during this period they were fed 3 to 5 percent of their weight daily with Fradaneh Company trout feed. Construction and commissioning of recirculation system In construction of recirculation systems 250-liter fiberglass tanks were used as fish rearing reservoirs liter plastic barrels were used as biofilter reservoirs and five-inch pieces of plastic pipe with a diameter of 2.5 cm were placed in the biofilter reservoirs as a media. Each system consisted of three tanks for fish rearing and one biofilter reservoir that connected to each other and the reservoirs water after circulating in system by a pump from the latest reservoir was transferred to biofilter reservoir. There was foam on top of the biofilter reservoirs in order to prevent the entrance of fish wastes and remaining foods into the reservoir. Measurement of Water quality parameters During experiment period, some of the water factors such as total ammonia, nitrite and nitrate once every 15 days before daily feeding were measured by a photometer instrument (GmBh and Tintometer: Model pc22). Moreover, other factors such as ph, EC and DO were measured by commercial kits (Darmstadt, Merck; Aquamerck, Australia) conductivity meter; tps; Aqua-CP instrument (WTW oxi33 oi). Measurement of stress factors During the experiment period, at the end of each month along with biometric operation, three fish from each treatment were randomly selected and blood sampling was done from caudal peduncle by 2-ml heparinized syringe (Mokhayer, 2006). Then the blood sample was centrifuged (3000 rpm, 7 minutes) and plasma was separated, after that cortisol and glucose were measured by photometric method and Pars Azmoon kits. RESULTS The results showed that the treatment containing 100 fish per m 3 had a positive effect on electrical conductivity and increased it but the treatment caused decreasing in Alkalinity and dissolved oxygen (P<0.05). The total ammonia increased in all the treatments from the beginning of the experiment until day 60 (P<0.05), then this factor started a downward trend in the treatments containing 75 and 100 fish per m 3 (P<0.05) and in the treatment containing 50 fish per m 3, the downward trend started from day 75 after the experiment (P<0.05) was started. But nitrite in all the experimental groups increased 2 weeks later after ammonia increasing until day 75 (P<0.05) and then nitrite decreased in the treatments containing 75 and 100 fish per m 3 (P<0.05) but the factor had no significant decrease during the experimental period in the treatment containing 50 fish per m 3 (P>0.05). Finally, nitrate in the treatment containing 100 fish per m 3 from day 15 and in the treatments containing 50 and 75 fish per m 3 from day 30 started an increasing trend (P<0.05) and the trend continued until the end of the experiment period.

3 Table 1. changes in the parameters of alkalinity, electrical conductivity and dissolved oxygen in the various densities of P. brachypomus in recirculation system. Treatments (fish per m 3 ) Factor A (50) B (75) C (100) Alkalinity 7.61±0.035b 7.57±0.036b 7.38±0.060a Electrical conductivity (EC) 539±21.93a 563±9.165ab 593±10.58b Dissolved oxygen (DO) 6.33±0.078b 6.18±0.053ab 6.03±0.225a Statistical significance is denoted by different letters. The results of stress factors measurement of fish blood showed that changes in water quality parameters in the treatments containing 50 and 75 fish per m3 had no effect on the cortisol and glucose indices in fish blood during the experiment. Table 2. changes in the parameters of total ammonia, nitrite and nitrate in the various densities of P. brachypomus in recirculation system. Treatments (fish per m 3 ) Factor Ammonia (TNT) (mg/l) Nitrite (NO2) (mg/l) Nitrate (NO3) (mg/l) Time (Day) A (50) B (75) C (100) ±0.012a 0.050±0.010a 0.050±0.017a ±0.012a 0.100±0.020a 0.107±0.029b ±0.026b 0.480±0.030b 1.090±0.026c ±0.088d 1.523±0.040e 1.850±0.020f ±0.070d 1.290±0.036d 1.483±0.035e ±0.026c 0.850±0.044c 1.230±0.050d ±0.000a 0.110±0.030a 0.120±0.026a ±0.021a 0.390±0.026b 0.440±0.044b ±0.017b 0.780±0.034c 0.920±0.095c ±0.035c 1.290±0.044d 1.710±0.062d ±0.040d 1.600±0.080e 2.190±0.071e ±0.053d 1.320±0.070d 1.767±0.070d ±0.023a 0.093±0.023a 0.200±0.044a ±0.265b 1.833±0.231b 2.033±0.153a ±0.265c 5.500±0.436c 8.300±0.265b ±0.458d 9.600±1.153d ±0.600c ±0.755e ±0.361e ±0.954d ±1.308f ±1.357f ±2.862e Statistical significance is denoted by different letters. In the treatment containing 100 fish per m 3, changes in water quality parameters had no effect on stress parameters until the end second month (P>0.05) but cortisol and consequently glucose in the treatment had a significant increase in sampling which was done in the third month (P< 0.05). From the relationship between water quality parameters and stress factors of fish blood it can be deduced that in this experiment reduction in alkalinity and increasing in nitrite and nitrate have been effective in increasing of stress factors of fish (P<0.05) and reduction of dissolved oxygen and increasing of electrical conductivity and total ammonia had no effect on stress factors separately (P>0.05).

4 Figure 1. changers cortisol in fishes blood in the various treatments and times (* indicate significant different with previous sampling). Figure 2. Changes in glucose in fish blood in the various treatments and times (* indicate significant different with previous sampling). DISCUSSION Stress in fish is a physiological change that occurs when fish expose to physiological, chemical and biological changes (Wedemeyer et al., 1990), and in this regard, water quality is one of the most important factors for fish farming that it is possible through its proper management reduced the time when fish expose to stress condition in recirculation systems (Wedemeyer, 1996) because environment life and condition of fish farming effect on content and blood metabolites (Bullis, 1993). The main feature of the stress response in fishes is in the activation of hypothalamic pituitary internal (HPI) axis that leading to increasing in cortisol secretion (Bernier and Peter, 2000), this part is a general response that during that cortisol activates the energy resourses and also help to ion balance keeping (Sattari, 2002) but if the

5 amounts of cortisol in the blood plasma increase chronically, instead of adaptive role it will have destructive effects on fish (Maule et al., 1988; VanWeed and Komen, 1998; Haddy and Pankhurst, 1999). Cortisol secretion increases in result of a variety of stressful stimuli such as lack of water having an appropriate quality, manipulation, closed condition, pollutants and water acidification (Barton and Peter, 1980). The factors that control the cortisol secretion are complex, in addition to ACTH, a variety of hormones such as -MSH, growth Hormone, thyroxine, Angiotensin, arginine vasotosyn, catecholimines, ANP, and ureotensin can produce steroids and a range of various factors have an interaction to increase in cortisol (Sattari, 2002). The factors produced by the immune system also modify cortisol secretion that this act is done directly or indirectly through influence on different components of HPI axis. In response to stress due to poor water quality, hypothalamus stimulates the pituitary in order to release ACTH and ACTH that reaches to anterior part of kidney through blood circulation and there, ACTH stimulates inter renal cells in order to cortisol production and other glucocorticoids hormones (Mommsen et al., 1999). Also parasympathetic nervous system stimulates the chromafin tissue in the anterior part of kidney in order to production of catecholamine hormones such as adrenaline (Evans, 1998). Both of the groups of mentioned hormones cause cardiovascular changes such as increasing in blood flow and blood pressure and increasing in oxygen consumption and also chemical changes like increasing in glucose level (Barton and Iwama, 1991). These changes are adaptive and fish can compensate the effects and also cause increasing in possibility of fish survival, Nevertheless, if the stress is severe and chronic finally a proper adaptation will not occur with the new condition and physiological condition of fish will decline (Wedemeyer, 1996). Due to the increased use of recirculation systems for fish farming and due to such systems are prone to decreasing in quality of water physicochemical factors and also according to the fact that if fish energy waste in response to poor water quality, there will be economic losses, so it is important to understand the relationship between water quality factors and fish stress indices, therefore, studies in this field are growing (Huntingford et al., 2006; Sinha et al., 2012). In the present study, although all the water factors that were measured affected by density, just decrease in alkalinity and increase in nitrite and nitrate were similar to increase in stress indices of cortisol and glucose and decreasing in dissolved oxygen and increase in electrical conductivity and total ammonia separately had no effect on stress indices while some studies in this regard showed a non-similar results (Abbas and Authman, 2009; Ravindrababu and Neeraja, 2012), this could be due to different condition of farming, specie, size and experimented fish age in various studies (Bullis, 1993). Thus, according to this study, in order to understand the relationships between water quality parameters and stress factors in various species in different conditions separate experiments are needed. REFERENCES Abbas HHH, Authman MMN Effect of accumulated on some physiological parameters and oxidative stress indicators in tilapia fish (Oreochromis spp). American- Eurasian J. Agric & Environ. Sci 5 (2), Barton BA, Iwama GK Physiological changes in fish from stress in aquaculture with emphasis on the response and effects of corticosteroids. Annual Review of Fish Disease 1, Barton BA, Peter RE, Paulencu CR Plasma cortisol levels of fingerling rainbow trout (Oncorhynchus mykiss) at rest and subjected to handling, confinement, transport, and stocking. Journal of Fish. Aquat. Sci 37, Barton BA Stress in finfish: Past, present, and future a historical perspective. In Iwama GK, Pickering AD, Sumpter JP, Schreck CB (eds). Fish stress and health in aquaculture. Cambridge University Press, Cambridge. pp Barton BA Stress in fish: a diversity of responses with particular reference to changes in circulating corticosteriods. Integ. Comp. Biol 42, Bernier NJ, Peter RE The hypothalamic pituitary interrenal axis and the control of food intake in teleost fish. Journal of Comparative biochemistry and physiology 129, Bullis RA Clinical Pathology of Temperate Freshwater and Estuaries Fish. In: Fish Medicine, Stoskopf, Evans DH The physiology of fishes, second Ed., CRC Press, New York, 519 pp. Haddy JA, Pankhurst NW Stress-induced changes in concentrations of plasma sex steroids in black bream. Journal of Fish. Biol 55, Huntingford FA, Adams C, Braithwaite VA, Kadri S, Pottinger TS, Sandoe P, Turbull JF Current issues in fish welfare. Journal of fish Biology 68, Maule AG, Tripp RA, Kaattari SL, Schreck CB Stress alters immune function and disease resistance in Chinook salmon (Oncorhynchus tshawytscha). Journal of Endocrinol 120, Mokhayer B Disease of culture fishes. Tehran university,428 pp. (In Persian). Mommsen TP, Vijayan MM, Moon TW Cortisol in teleosts: dynamics, mechanisms of action, and metabolic regulation. 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6 Ravindrababu G, Neeraja P Histological changes in certain tissues of fish on ambient ammonia stress and post ammonia state. Journal of Advanced biological research 2, Sanni S, Forsberg OI Modeling ph and carbon dioxide in single-pass seawater aquaculture systems. Aquaculture Engineering 22(1-2), Sattari M Ichthyology 1 (Anatomy and Physiology). Naghsh Mehr Press. Tehran, 659 p. (In Persian). Schreck CB, Li HW Performance capacity of fish: stress and water quality. In:Brune DE, Tomasso, JR. (Eds.), Aquaculture and Water Quality. Advances in World Aquaculture, vol. 3. Pub.Wld. Aquacult. Soc, pp Sinha AK, Dirix M, Chan LP, Liew HJ, Kumar V, Blust R, De Boeck G Expression pattern of potential biomarker genes related to growth, ion regulation and stress in response to ammonia exposure, food deprivation and exercise in common carp (Cyprinus carpio). Aquat toxicol 122, Timmons MB, Ebeling JM, Wheaton FW, Summerfelt ST, Vinci BJ Recirculating Aquaculture Systems. NRAC Publication no , Cayuga Aqua Ventures, Ithaca, NY, 650 pp. Turnbull JF Current issues in fish welfare. Journal of Fish Biology 68, VanWeerd JH, Komen J The effects of chronic stress on growth in fish: a critical appraisal. Comp. Biochem. Physiol 120, Vijayan MM, Moon TW Acute handling stress alters hepatic glycogen metabolism in fooddeprived rainbow trout (Oncorhynchus mykiss). Can. J. Fish. Aquat. Sci 49, Vijayan MM, Moon TW The stress response and the plasma disappearance of corticosteroid and glucose in a marine teleost, the sea raven. Can. J. Zool 72, Wedemeyer GA, Barton B, McLeay DJ Stress and acclimation. In: Schreck CB, Moyle PB (eds). Methods for Fish Biology. Bethesda: American Fisheries Society, p Wedemeyer GA Physiology of fish in intensive culture systems, Chapman and Hall, New York, 302 pp. Wedemeyer GA Effects of rearing conditions on the health and physiological quality of fish in intensive culture. In: Iwama GK, Pickering AD, Sumpter JP, Schreck CB. (eds.), Fish Stress and Health in Aquaculture. Soc. Exp. Biol. Semin. Ser. 62. Cambridge University Press, UK, pp

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