International Journal of Aquatic Biology (2014) 2(5): 275-285 E-ISSN: 2322-5270; P-ISSN: 2383-0956 Journal homepage: www.NPAJournals.com © 2014 NPAJournals. All rights reserved Original Article Effects of dietary administration of yarrow extract on growth performance and blood biochemical parameters of rainbow trout (Oncorhynchus mykiss) Mahmoud Nafisi Bahabadi1, Mahdi Banaee2*,1Marzieh Taghiyan1, Behzad Nematdoust Haghi2 1Aquaculture Department, Faculty of Agriculture and Natural Resources, Persian Gulf University, Bushehr, Iran. 2Aquaculture Department, Natural Resource Faculity, Behbahan Khatam Alanbia University of Technology, Behbahan, Iran. Article history: Received 4 June 2014 Accepted 24 August 2014 Available online 2 5 October 2014 Keywords: Medicinal plants Yarrow extract Rainbow trout Biochemical factors Growth performance Abstract: The present study was conducted to investigate the clinical effects and possible side effects of yarrow extract (Achillea millefolium L.) as feed additive on biochemical blood parameters and growth performance of rainbow trout (Oncorhynchus mykiss). Fishes were treated with 0 (control), 0.1, 0.5 and 1% of yarrow extract for 30 days. Plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), creatine kinase (CK), peroxidase activity, total complement and lysozyme activity, glucose, total protein, triglyceride and cholesterol were measured after 15 and 30 days of yarrow treatment. There were no significant changes in the lysozyme activity and glucose levels. Total protein and globulin levels were significantly higher in the fish fed with diets enriched with 1% yarrow extract on day 30. Triglyceride and cholesterol levels was significantly decreased in the fish fed with diets containing 0.5% and 1% yarrow extract on day 30 (P<0.05). LDH, CK and peroxidase activities in the fish fed with diets having 1% yarrow extract were significantly decreased at the end of the experiment (P<0.05). In contrast, a significant increase in AST, ALP and total complement activity was observed in the fish fed with 1% yarrow extract diet, on day 15 (P<0.05). The weight gain and specific growth rate increased and food conversion ratio decreased in in the fish fed 1% yarrow extract on day 30. Condition factor in the fish fed with yarrow extract was significantly higher than control group on 30 day. In conclusion, on the basis of these results, oral administration of yarrow extract up to 0.5% have not side effect on blood biochemical and clinical parameters of fishes. However, oral administration of 1% of yarrow extract caused cytotoxicity and modifications in blood biochemical parameters of fish. Introduction Compared with chemical drugs, the medicinal plants have effective agents and other compounds which accelerate gastrointestinal absorption process, improve the therapeutic effects and reduce side effects and drug toxicity (Platel et al., 2002). Although the results reported by laboratory animals evidence beneficial effects in the application of medicinal plants in treatment and prevention of disease in fish (Sivaram et al., 2004; Rao et al., 2006; Divyagnaneswari et al., 2007), the contradictory effects of some medicinal plants has decelerated using them for fish in a commercial scale. The lack * Corresponding author: Mahdi Banaee E-mail address: Mahdibanaee@yahoo.com of knowledge about the biological effects of compounds in medicinal plants' extracts on the individual characteristics of different fish species are the main problems in application of these compounds (Banaee, 2010). Therefore, study of the effects of herbal derivatives as a drug on the health of fish may increase our knowledge which can favour the use of certain compound for disease treatment (Banaee et al., 2011). Yarrow (Achillea sp.) has an important role in medicinal plants derived from its antioxidant (Yakhkeshi et al., 2012), analgesic, anti-bacterial (Mazandarani et al., 2007), anti-parasitic (Khalili et 276 International Journal of Aquatic Biology (2014) 2(5): 275-285 al., 2011), anti-fungal (Ayatollahi Mousavi et al., 1996), anti-inflammatory (Benedek et al., 2007), anti-hypertensive and lipid-lowering (Asgary et al., 2000), anticonvulsant and antispasmodic (Lemmens-Gruber et al., 2006), antitumor (Csupor- Löffler et al., 2009), anti-ulcer skin (Cavalcanti et al., 2006), and antiarrhythmic (Asgary et al., 2000) properties. The antibacterial properties of yarrow extract are attributed to flavonoids, saponins, and terpens (Table 1) (Mothana et al., 2009). Moreover, the administration of medicinal plants with antibiotic properties can change intestinal bacterial flora, increase growth and improve carcass quality (Tekeli et al., 2008). Therefore, the present study was carried out to determine whether yarrow (Achillea millefolium L.) would influence on the blood biochemical parameters, growth performance and immune parameters of rainbow trout. Materials and Methods Dried yarrow extract preparation: Yarrow powder was extracted with hydro-alcoholic mixture (ethanol 70% and water in 1:1 proportion) at room temperature by cold maceration method (Asgary et al., 2003). Then, this extract was filtered through Whatman filter paper. Finally, the extract was concentrated in an incubator at 50°C obtaining the dry extract. Animal treatments: One hundred and twenty rainbow trout (with average weight and length of Flavonoid Essential oil Cyaroside Tricyclene n-heptanol Cosmosiin α-pinene 3-octanone Luteolin β-ocimene δ-3-carene Apigenin γ-terpinene α-terpinene Centaureidin Terpinolene p-cymene Quercetin Linalool Pinocarvone 3´-methoxy luteolin Trans-thujone Borneol Luteolin 7-O-glucoside Allo-ocimene α-copaene 5-hydroxy 3´,4´,6´,7´-tetra methoxy flavone Terpin-1-ol Geranyl acetate Camphor Longifolene Salvigenin (Z)-tagetone α-gurjunene Galangin Isoborneol α-guaiene Eupatilin α-terpineol n-pentadecane α-himachalene α-cadinene γ-gurjunene γ-eudesmol β-chamigrene α-eudesmol γ-muurolene Bornyl acetate β-selinene Neryl acetate Viridiflorene α-cyclogeraniol Cis-pinocarveol β-calacorene Isobornylformate (Z)-cubenol α-terpinyl acetate β-bisabolol Caryophyllene alcohol Phytol (E)-cinnamaldehyde Nootkatin Patchouli alcohol Occidol acetate α-bisabolene oxide A Abietadiene (E,E)-farnesol n-heneicosane Chamazulene Methyl octadecanoate Table 1. Phytochemical compounds isolated from the dried yarrow extract adopted from Saeidnia et al. (2011) and Ebrahimi et al. (2012). 277 Nafisi Bahabadi et al/ Effects of yarrow extract on rainbow trout 146.9 ± 18.1 g; 25.9 ± 2.6 cm, respectively) were randomly introduced into 4 treatments, 3 replicates with 10 fish in the 12 fibreglass tanks (200 L) with semi-closed water recirculating systems for two weeks to acclimate to the laboratory conditions (15 ± 2°C; pH: 7.4 ± 0.2; photoperiod: 16L: 8D; 20% water exchange rate/day) prior to experiments. During acclimation and experimental periods, fish were fed with prepared pellets according to commercial formulations at 2% of their body weight twice a day (Hinshaw, 1999). The enriched diet was prepared with formulated fish feed obtained from Chineh Company, Karaj, Iran (Table 2) and yarrow extract according to the method developed by Banaee et al. (2011). Each supplemented diet was mixed with distilled water until obtaining a homogenous mixture. This mixture was passed through a meat grinder, producing extruded string shapes, which were dried in an oven at 55°C for 12 hrs and then broken to produce pellets with approximate 10 mm length. The prepared pellets were packed and stored at -20°C in a freezer until be used. The control diet was prepared using the same process, although no supplement was added. In day of 15 and 30 of the experiment, 12 fish from each treatment were randomly sampled and anesthetized with clove powder solution (200 mg/L). Then, the blood of fish from each experimental and control groups were taken by puncturing of the caudal vessels with a syringe and the blood was stored in sterilized glass vials containing the anticoagulant 1% EDTA at 4°C. The blood was centrifuged for 15 min at 4000 rpm. The contents of digestive tract, nutritional status (gastro-intestine- somatic index) and clinical symptoms such as changes in the size of gall-bladder and liver (hepato- somatic index) as well as feeding behavior of treated fish were evaluated. Biochemical Analysis: Biochemical parameters of plasma including glucose, total protein, albumin, globulin, cholesterol and triglyceride; aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), alkaline phosphatase (ALP) and creatine kinase (CK) were analyzed using the biochemical diagnostic kit (Pars- Azemon Company, Karaj, Iran). Alternative complement activity (ACH50) was evaluated based on Yano (1992) using rabbit red blood cells (RaRBC). In this method, the absorbance of the haemolysate was measured at 414 nm against distilled water to acquire the 100% lysis value. The volume of plasma producing 50% haemolysis (ACH50) was determined. Lysozyme activity was determined by the lysis of bacterial cells of Micrococcus lysodeikticus (Lange et al., 2001). The decrease in turbidity at 570 nm after 0.5 min and 4.5 min was recorded by spectrophotometer. Lysozyme of sample was calibrated using a standard curve determined with hen’s egg white lysozyme (Sigma) in PBS. The total peroxidase activity was determined according to Cuesta et al. (2007) with modification by measuring the optical density of 3,3′,5,5′– tetramethylbenzidine hydrochloride oxidation products during particular intervals of time at 450 nm. All biochemical parameters were measured in duplicates by UV/Vis Spectrophotometer (model Unico 2100). Growth performance: The growth parameters including weight gain percentage, specific growth rate, and feed conversion ratio and condition factor were calculated using the following formulas, on day15 and 30 day. Weight gain (%) = Final weight − Initial weight Initial weight × 100 Specific growth rate (SGR%) = [(Ln (final body weight)-Ln (initial body weight))/(Experimental periods)]×100 Nutrient composition Reference diet Dry material 92 Metabolize energy (Kcal/g) 350 Crud protein 40 Ether extract (lipid) 10.5 Ash 7.9 Crude fiber 5.8 Carbohydrate 27 Table 2. Nutrient composition of the commercial diets (Chineh Company, Iran). 278 International Journal of Aquatic Biology (2014) 2(5): 275-285 Feed conversion ratio (FCR) = Feed intake (g) Wet weight gain (g) Condition factor (CF) = Weight (g) (Length (cm))3 × 100 Statistical analysis: A significant difference in the biochemical parameters of specimens treated with the different concentrations of A. millefolium extracts was examined using one-way ANOVA. All the data were examined for normality (Kolmogorov- Smirnov test). The means were compared by Duncan’s test and a P<0.05 was considered statistically significant. Statistical analyses were performed using SPSS (IBM, Release 19) software. Data are presented as mean ± SD. Results The hepato-somatic index (IH) was significantly decreased in fish fed with a diet containing 1% yarrow extract. No significant changes were observed during the test in spleen-somatic index (IS) and gastro-intestine-somatic index (IG) in the fish fed with diets containing yarrow extract when compared with control group. The contents of the digestive system of fish as satiety index indicated that there were no significant difference between amounts of feed was fed by experimental fish and control group (Fig.1a, b, c). Significant changes in the WG and SGR was observed in the fish fed with a diet containing 1% yarrow extract during the experiment. The FCR was significantly decreased in fish with diets having 1% yarrow extract. The CF in the fish fed with yarrow extract was significantly higher than the control group at the end of experiment (Table 3). There was no significant difference in glucose levels between fish fed yarrow extract and control group during experimental period. Although, oral administration of 1% yarrow extract resulted in a significant increase in total protein levels, a significant decrease in the albumin levels was observed in plasma of fish fed with 0.1% yarrow extract on day 30. Supplementing feed with 0.1 and 1% yarrow extract resulted in significantly increased in globulin levels on day 30 (P<0.05). Triglyceride levels in the plasma of the fish fed with diets having 0.5% and 1% yarrow extract showed a significant decrease during the test. Cholesterol levels Figure 1. Effects of different concentrations of yarrow extract as feed additive on (a) percentage ratio of spleen to total weight as spleen-somatic index (IS), (b) liver weight to total weight as hepato-somatic index (IH) and (c) gastrointestinal tract weight to total weight (IG) as a satiety index or gastro-intestine-somatic index. Significant differences between treatment and control groups were represented by alphabets (one-way ANOVA, P<0.05). Values represent mean ± S.D. 279 Nafisi Bahabadi et al/ Effects of yarrow extract on rainbow trout significantly decreased in plasma of fish fed with diets having 0.5% and 1% yarrow extract on day 30 of the test (P<0.05), (Table 4). There was a significant increase in the total complement activity in plasma of fish fed with food enriched with 1% of yarrow extract on day 30 of trial (P<0.05). No significant changes were observed in the ACH50 activity on day 15 (Table 4). The activity of AST significantly decreased in plasma of fish fed with 0.1% and 0.5% yarrow extract on day 30 of experiment (P<0.05). However, AST activity in plasma of the fish fed with diets having 1% yarrow extract significantly increased on day 15 of test (P<0.05). The activity of ALT significantly increased in plasma of fish fed with 0.5 and 0.1% yarrow extract on 15 and 30 days when compared with control group, respectively. LDH activity in plasma of the fish fed with diets enriched with 0.1% and 1% yarrow extract was significantly lower than control group on day 30 of the experiment. The activity of ALP significantly increased in plasma of fish fed with 1% yarrow extract on day 15 of experiment. The activity of CPK significantly decreased in plasma of fish fed with 1% yarrow extract during experimental period (P< 0.05), (Table 5). A significant decrease in the peroxidase activity was observed in plasma of fish fed with 0.5% and 1% yarrow extract on day 15 (P< 0.05). Also, there was no significant change in peroxidase activity on day 30 of test. No significant changes in lysozyme activity in plasma of fish fed with yarrow extract supplement were observed when compared with control group during experimental periods (P> 0.05), (Table 5). Discussion In this study, rainbow trout were fed with diets enriched with 0.1%, 0.5%, and 1% yarrow extract for 30 days. During the experimental periods, no mortality or changes in the appetite of the fish were observed. Platel et al. (2002) evidenced the favorable effect of medicinal plants on digestion and a stimulating effect on bile secretion and the activity of pancreatic enzymes. Moreover, adding plants extracts to the diet can affect the fish ability to find Growth Performance YE% of g/kg Experimental periods 15th day 30th day Weight gain (%) 0.0 27.45±7.45a 56.63±8.09a 0.1% 35.80±11.85ab 70.81±18.77b 0.5% 28.33±6.53a 67.23±11.43ab 1% 39.57±8.74b 69.79±9.36b Specific growth rate (%/day) 0.0 1.61±0.38a 1.49±0.17a 0.1% 2.02±0.59ab 1.77±0.37b 0.5% 1.66±0.33a 1.71±0.23ab 1% 2.21±0.42b 1.76±0.18b Food conversion rate 0.0 1.73±0.54b 1.59±0.24b 0.1% 1.40±0.62ab 1.33±0.37ab 0.5% 1.63±0.34ab 1.35±0.27ab 1% 1.18±0.33a 1.28±0.17a Condition factor 0.0 0.85±0.06a 0.89±0.06a 0.1% 0.92±0.07a 1.00±0.09b 0.5% 0.93±0.13a 0.98±0.06b 1% 0.93±0.0.9a 0.98±0.08b Effects of different concentrations of yarrow extract on growth index were analyzed using a one-way ANOVA. Significant differences between treatment and control groups were represented by alphabets (P<0.05). Values represent mean ± SD. Table 3. Growth performance of rainbow trout fed with yarrow extract as a supplement. 280 International Journal of Aquatic Biology (2014) 2(5): 275-285 food by stimulating their sense of smell and encourage them to eat more (Adams, 2005). Our results indicate that feeding of fish with 1% yarrow extract significantly improved growth performance. Some compounds in medicinal plants extracts including bioflavonoids can positively induce effects on growth performance and general health of fish (Yilmaz et al., 2006). Similar results are found in cichlid, Cryptoheros nigrofasciatus, (Cek et al. 2007); red seabream, Pagrus major (Ji et al. 2007); and rainbow trout (Bohlouli Oskoii et al., 2012) which were fed with diets supplemented with medicinal plants extracts. Although, the results indicate that administration period of yarrow extract had a significant effect on glucose levels in plasma, hypoglycemic effect of yarrow on rats with diabetes mellitus indicates this plant's effects on against in regulating blood sugar in these animals (Sadeghi et al., 2009). These results may be attributed to the hypoglycemic activity of Biochemical Parameters YE% of g/kg Experimental periods 30th day 15th day Glucose (mg/dL) 0.0 % 74.64±26.41a 89.58±4.42a 0.1% 64.53±11.46a 83.46±7.41a 0.5% 73.32±22.68a 87.20±8.30a 1% 62.59±13.17a 83.08±5.56a Total protein (g/dL) 0.0 % 4.54±0.55a 4.77±0.87a 0.1% 4.31±0.42a 4.52±0.59a 0.5% 4.32±0.48a 4.45±0.41a 1% 4.84±0.47b 4.37±0.91a Albumin (g/dL) 0.0 % 3.00±0.40b 2.84±0.18a 0.1% 2.36±0.56a 2.89±0.09a 0.5% 3.00±0.51b 2.88±0.48a 1% 2.77±0.62ab 2.88±0.11a Globulin (g/dL) 0.0 % 1.53±0.36a 1.93±0.82a 0.1% 1.95±0.67b 1.63±0.61a 0.5% 1.32±0.35a 1.56±0.39a 1% 2.07±0.43b 1.49±0.90a Cholesterol (mg/dL) 0.0 % 222.11±41.41b 220.77±29.07ab 0.1% 226.75±34.86b 258.18±52.80b 0.5% 182.22±36.49a 178.16±42.49a 1% 168.43±38.39a 191.25±49.81a Triglyceride (mg/dL) 0.0 % 167.38±39.68b 166.97±33.09b 0.1% 125.01±38.83a 169.77±12.43b 0.5% 104.04±40.61a 129.60±17.07a 1% 114.29±28.02a 126.02±26.34a ACH50 (U/mL) 0.0 % 311.89±9.05a 321.44±7.86a 0.1% 321.89±43.96a 315.67±11.05a 0.5% 344.45±19.18a 314.34±10.06a 1% 344.45±46.22a 366.00±30.56b Effects of different concentrations of yarrow extract on biochemical parameters. Significant differences between treatment and control groups were represented by alphabets (one-way ANOVA, P<0.05). Values represent mean ± SD. Table 4. Plasma biochemical parameters in rainbow trout fed with yarrow extract as a supplement. 281 Nafisi Bahabadi et al/ Effects of yarrow extract on rainbow trout herbal extracts to increase the level of plasma insulin and improvement of peripheral metabolism of glucose (Awad, 2010) or may be attributed to the activation of glycogen synthesis and healthy hepatic function (Ji et al., 2007). Decreased glucose levels in the blood of rainbow trout fed with silymarin extract (Banaee et al., 2011) and African catfish fed with onion and garlic extracts are reported (Al-Salahy, 2002). Albumin and globulin make up most of the protein within the body and are measured in the total protein of the plasma. Total protein, albumin and globulin tests are used to monitor the course of diseases in immune disorders, liver dysfunction and impaired kidney activity. At the end of the test, the levels of albumin in plasma of the fish fed with diets containing 0.1% yarrow significantly decreased. The 0.1% of yarrow extract was able to significantly increase the total protein levels on day 30. Oral administration of 0.1 and 1 % yarrow extract significantly increased the globulin levels in plasma of fish on day 30. Although an increased level of total protein, albumin and globulin in plasma of the fish fed with diets enriched with Echinacea purpurea and Silybum marianum was reported by Bohlouli Oskoii et al. (2012) and Banaee et al. (2011), no significant changes are reported in the levels of albumin and total protein in plasma of fish fed with diets enriched with onion and garlic extract (Al- Salahy, 2002; Naeiji et al., 2013).. Quercetin is the most important flavonoid in yarrow (Saednia et al., 2009) that prevents the biosynthesis of cholesterol by inhibiting the activity of fatty acid synthesis (Yamamoto and Oue, 2006). Therefore, the Biochemical Parameters YE% of g/kg Experimental periods 15th day 30th day AST (U/L) 0.0 % 87.27±12.65a 87.49±32.49b 0.1% 97.49±13.94a 63.23±16.61a 0.5% 96.75±16.18a 63.15±10.65a 1% 121.89±36.22b 69.91±13.59ab ALT (U/L) 0.0 % 12.86±3.67a 17.27±2.65a 0.1% 12.57±4.39a 23.60±8.39b 0.5% 19.11±5.52b 17.64±4.44a 1% 12.79±3.87a 18.08±4.45a LDH (U/L) 0.0 % 808.89±103.17a 735.60±237.85b 0.1% 884.44±169.41a 523.05±203.21a 0.5% 882.22±116.76a 700.57±251.85ab 1% 890.37±205.86a 493.96±94.42a CPK (U/L) 0.0 % 177.31±23.81b 207.27±27.47b 0.1% 167.11±12.29ab 199.15±27.53b 0.5% 160.34±31.61ab 199.99±37.76b 1% 150.85±28.84a 151.19±31.27a ALP (U/L) 0.0 % 18.89±5.59a 25.93±4.45a 0.1% 25.12±9.63a 23.69±12.64a 0.5% 34.41±20.40ab 21.95±4.12a 1% 45.44±25.92b 20.22±11.65a Peroxidase (U/mL) 0.0 % 138.11±10.78b 139.44±10.49a 0.1% 129.67±11.68ab 126.11±11.21a 0.5% 120.00±10.10a 119.56±9.41a 1% 120.44±9.38a 163.44±86.12a Lysozyme (U/mL) 0.0 % 123.68±8.38a 120.67±10.83a 0.1% 124.78±9.13a 125.11±11.95a 0.5% 128.22±11.10a 126.00±14.01a 1% 129.44±10.10a 124.22±18.46a Effects of different concentrations of yarrow extract on biochemical parameters (Enzyme activity). Significant differences between treatment and control groups were represented by alphabets (one- way ANOVA, P<0.05). Values represent mean ± SD. Table 5. Plasma enzymatic activities in plasma of rainbow trout fed with yarrow extract as supplement. 282 International Journal of Aquatic Biology (2014) 2(5): 275-285 decreased cholesterol levels in the blood of fish fed with 0.5 and 1% yarrow extract at the end of the test, and decreased levels of triglycerides in plasma of the fish fed with diets having 0.5% and 1% yarrow extract during the test may be attributed to the influence of quercetine on the synthesis of cholesterol. The influence of long term use of yarrow in decreasing triglyceride, total cholesterol and low- density lipoprotein (LDL) and increasing the high density lipoprotein (HDL) in the blood of human volunteers verifies this issue (Asgary et al., 2000). The cholesterol synthesized in liver transfers to other tissues of the body through LDL, but HDL transports the cholesterol of peripheral tissues to liver. So, the increased excretion of cholesterol through bile (Asgary et al., 2000) decreases the cholesterol level in blood of the fish fed with yarrow. Decreases in cholesterol and triglyceride levels are also reported in blood of rainbow trout and catfish respectively fed with silymarin extract (Banaee et al., 2011) and onion and garlic extracts (Al-Salahy, 2002). The complement system is an essential and effective part of the innate immune system. It can rapidly distinguish and opsonize bacteria for phagocytosis by specialized phagocytes or destroy them directly by membrane disorder (Ahmadi et al., 2014). The increase in the complement activity (ACH50) observed in plasma of fish fed with diets enriched with 1% of yarrow extract on day 15 may help to identify and eliminate bacterial agents by phagocytosis. Increases in the total complement activity were reported in fish fed with a diet enriched with Punica granatum, Chrysanthemum cinerariaefolium and Zanthoxylum schinifolium extracts (Harikrishnan et al., 2010), S. marianum (Ahmadi et al., 2012) and Nasturtium nasturtium extracts (Asadi et al., 2012). AST, ALT, LDH, ALP and CK are found in various tissues. When diseases or injuries affect these tissues, the cells are destroyed and these enzymes are released into plasma (Banaee et al., 2011). Although, oral administration of 0.1 and 0.5% yarrow extract resulted in a significant decrease in the activity of AST on day 30, a significant increase in AST activity in plasma of fish fed with enriched with 1% yarrow extract on day 15. Treatment with 0.5 % yarrow extract resulted in a significant increase in AST activity on day 15. Our results show that ALP activity significantly increased in the plasma of fish fed with enrich diet with 1% yarrow extract when compared with the control group at day 15, no significant changes were observed in ALP activity at day 30. LDH measurement is used to detect tissue disorders and as an aid in the diagnosis of tissue damage. The LDH activity also significantly decreased after the oral administration of 0.1 and 1% yarrow extract on day 30. A significant decrease were observed in the CK activity in plasma of common carp fed with enrich diet with 1% marshmallow flower extract when compared with the control group during experimental period. The most important of the biological mechanisms of flavonoids have been attributed to their antioxidant properties (Oteiza et al., 2005). Due to anti-radical and anti-oxidant properties of the yarrow extract, its administration might prevent lipid peroxidation of cell membranes and inhibit the release of foresaid enzymes into the plasma. Therefore, yarrow may be useful in treatment and prevention of diseases caused by oxidative stress derived from its rich content in flavonoids. A low concentration of silymarin (Banaee et al., 2011) and garlic (Al-Salahy, 2002; Naeiji et al. 2013) in diet of fish has been evidenced to regulate the plasma activities of AST, ALT, ALP, CK and LDH. More hydrophilic flavonoids can interact at the membrane surface through hydrogen bonding; can act to reduce the access of oxidants and per-oxidants and protect the structure and function of cellular membranes (Oteiza et al., 2005). Lysozymes are a family of enzymes with antibacterial activity characterized by the ability to damage the cell wall of bacteria (Ahmadi et al., 2014). Our results showed that fish fed with diets containing yarrow extract had no effects on lysozyme activity. Studies of Sivaram et al. (2004) showed that no significant changes were observed in serum lysozyme activity of cultured grouper, Epinephelus tauvina, fed with diets enriched with 283 Nafisi Bahabadi et al/ Effects of yarrow extract on rainbow trout Ocimum sanctum, Withania somnifera and Myristica fragrans extracts. Peroxidases play an important role in defense system against extracellular bacterial and parasitic pathogens. Myeloperoxidase and eosinophil peroxidase are important active peroxidases in immune system of fish and found in the cytoplasmic granules of neutrophils and eosinophil, respectively. If the leukocytes are stimulated, peroxidase activity will increase in plasma (Ahmadi et al., 2014). The results revealed that the administration of yarrow extract to fish significantly decrease the peroxidase activity. Antibacterial compounds that exist in the yarrow extract may be prevented stimulation of leukocytes. Ahmadi et al. (2012) and Asadi et al. (2012) found that the administration of S. marianum and N. nasturtium extract had no effects on peroxidase activity of fish. Since the change in blood biochemical parameters is a useful clinical tool to predict and monitor the health in organisms, these factors can also be used for determining drug safety. Preclinical research on the biochemical factors and growth performance as pharmacology indices of yarrow revealed that this herbal medicine may be useful by providing benefit in the treatment of some disease of aquatic animals. Therefore, in clinical studies should be focused on the medical effects of yarrow. Acknowledgements Special thanks are extended to research assistant of Faculty of Natural Resources, Persian Gulf University for their financial support. The authors are also grateful to the Dr. Antoni Sureda for their cooperation and guidance throughout the writing manuscript. Reference Adams C. (2005). Nutrition-based health. Feed internat. 2: 25-28. 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