Trigonella Foenum Graecum) and Liquorice (Glycyrrhiza Glabra) on Growth Performance, Antioxidant Activities and Stress Resistance in Oreochromis Niloticus Sarah A
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BENHA VETERINARY Basha MEDICAL et al.,2018 JOURNAL, BVMJ -V34(3):OL. 34, 355 N-O37. 3:1 355-370, AUGUST 2018 A SPECIAL ISSUE FOR THE 6TH SCIENTIFIC INTERNATIONAL CONFERENCE BENHA UNIVERSITY FACULTY OF VETERINARY MEDICINE Dietary effect of fenugreek (Trigonella foenum graecum) and liquorice (Glycyrrhiza glabra) on growth performance, antioxidant activities and stress resistance in Oreochromis niloticus Sarah A. Basha1, Eman A. Abd El-Gawad2, Ashraf A. Abd EL latif2 and Amany A. Abbass2* 1Directorate of Veterinary Medicine Gharbiya Governorate, Egypt. 2Department of Aquatic Animal Diseases and Management, Faculty of Veterinary Medicine, Benha University, Egypt. *Correspondence to: Amany A. Abbass. Email: [email protected] A B S T R A C T This work aimed to investigate the effect of dietary fenugreek seeds and liquorice roots on growth performance, immune status, antioxidant and stress resistance in Nile tilapia (Oreochromis niloticus). For this, five groups of fish each in duplicate were used, the 1st 2 groups were fed daily on experimental diets 1% and 2% (wt/wt) fenugreek; the 2nd 2 groups were received 1% and 2 % (wt/wt) liquorice and the remaining group was fed on control diet for 90 days. Growth performance (ABW, WG, WGR, SGR, L and LGR) and intestino-somatic index of fish fed 2% fenugreek supplemented diet were significantly increased (P < 0.05) compared to control. Hematological parameters (RBCs, WBCs, PCV, hemoglobin concentration and DLC) revealed significant increase in groups fed 2 % fenugreek and 2% liquorice. Serum total protein and globulin were significantly increased in groups received 1 and 2 % liquorice supplemented diets. Liver enzyme (ALT) in fish fed 1 and 2 % liquorice diets was lower than control. Blood cortisol and glucose showed significant decrease with 1 % fenugreek. Liquorice treated groups exhibited significant decrease in lactate dehydrogenase (LDH). Hepatic antioxidant enzymes [(SOD), (CAT), and (GSH)] showed significant increase with 2 % dietary fenugreek and 2% liquorice, while Malondialdehyde (MDA) was significantly decreased compared to control. Post feeding, sub-groups of fish were exposed to 6 days starvation and hypoxia stress and the results showed that, cortisol and glucose levels were significantly decreased in groups fed 2 % fenugreek and liquorice treated diets and the antioxidant enzymes SOD, CAT and GSH response were characterized by significant improvement with lower oxidative stress (MDA) level compared to control. Results suggested that dietary supplementation of fenugreek seeds and liquorice roots enhance growth performance and improve resistance against starvation and hypoxia in Oreochromis niloticus. Keywords: Growth, Fenugreek, Liquorice, Immunity, tilapia fry, stress. (http://www.bvmj.bu.edu.eg) (BVMJ-34(3): 355-370, 2018) 355 Basha et al.,2018 BVMJ-34(3): 355-371 1. Introduction Glabridin is a polyphenolic flavonoid Aquaculture is an important source for bioactive compound from liquorice has seafood than fisheries. Fish consumption is immunostimulants properties (Zore et al., reported to increase further in countries such 2008). This study aimed to evaluate the effects as Africa, Asia, America, and European of dietary fenugreek seeds and liquorice roots regions during 2010–2030 (Kobayashi et al., on the growth performance, Immuno- 2015). There are many management practices haematological parameters, antioxidant such as overcrowding, sudden changes in activities and resistance against starvation and temperature, poor water quality, low oxygen hypoxia stress in Oreochromis niloticus fry. level, bad handling and starvation which lead 2. Materials and methods to physiological disturbance, immune 2.1. Fish suppression and increase susceptibility to Nile tilapia, O. niloticus fry with an infectious diseases (Cabello, 2006; Quesada et average body weight 2±0.4 g was obtained al., 2013). The main method to control from fish farm at Kafr El Sheik province, diseases is antibiotics but it has disadvantages Egypt. Fish were stocked in well prepared such as accumulation of residues in fish tissue fiberglass tanks (750L) for 2 weeks or in the environment and drug resistant acclimation period and fed twice daily on pathogen (Santos and Ramos, 2016). basal diet (42 % crude protein). The health Although, vaccination is a good method of conditions of fish were examined for any disease prevention, but it is too expensive and diseases as described by Austin and Austin unpractical for wide use in fish farms (1989). The water temperature was adjusted to (Harikrishnan et al., 2011). Therefore, there is 25±2°C and dissolved oxygen was maintained a great attention for use the whole herbal at 5mg/L plants or its extracts as an alternative method due to its cheap price, provide better growth 2.2 Herbal plants for fish and protection against disease (Bahi et Herbal plants fenugreek (Trigonella al., 2017) and also as appetite stimulation, foenum graecum) seed sand liquorice antimicrobial, immune stimulation and stress (Glycyrrhiza glabra) roots were purchased resistant in fish (Chakraborty and Hancz., from local hypermarket in El-Gharbiya 2011). province, Egypt. The dried seeds of fenugreek Fenugreek seeds have antioxidant, and liquorice roots were grinded to fine immuno modulatory effect, anti-carcinogenic, powder by electrical blinder and preserved at o anti-ulcer due to their bioactive contents of 4 C in refrigerator till use. flavonoids such as: apigenin, kaempferol and 2.3. Preparation of experimental diets quercetin and saponins including diosgenin The experimental diets were prepared and yamogenin. (Olaiya and Soetan, 2014). from the following ingredients according to Enhancement of growth performance and fish NRC, (2011) as follow; rice bran 125 g , immune system after dietary administration of wheat bran 23 g, fish meal 300 g, yellow corn fenugreek seeds was recorded by Bahi et al., 150 g, soybean 350 g, glutean meal 25 g, (2017) and Guardiola et al., (2018). vegetable oil 25 g and vitamin premix 3g . Liquorice roots possess a considerable Five diets were prepared; first diet (basal diet number of properties including those that are 0%) was kept as control, second and third anti-inflammatory, anti-viral, and diets were incorporated with 1 and 2 % antimicrobial (Asl and Hosseinzadeh, 2008). 356 Dietary effect of fenugreek (Trigonella foenum graecum) and liquorice (Glycyrrhiza glabra) on growth performance, antioxidant activities and stress resistance in Oreochromis niloticus (wt/wt) fenugreek. The last two diets were hour using an oxygen meter to 3.5 mg/l and incorporated with 1 and 2 % (wt/wt) liquorice fish showed signs of hypoxia as surfacing with respectively. A Suitable amount of water rapid movement of operculum. containing the adequate amount of either 2.5. Blood and tissue sampling fenugreek seed powder or liquorice root power Blood and tissue samples were taken was mixed with all ingredients forming moist on day 90 post feeding, and after exposure to dough and pelleted. The obtained pellets were starvation stress for 6 days and to hypoxia allowed to dry at room temperature for 48 stress (stress experiment). days then packed in clean dry tightly closed Blood samples were withdrawn from the plastic containers and kept at 4ºC until use. caudal blood vessels of anesthetized fish using 2.4. Experimental design syringe moisten with EDTA anticoagulant for hematological parameters assay, and another The experiment was designed to include two trials: amount without anticoagulant to separate 2.4.1 Feeding experiment serum and stored at –80°C until used Liver tissues were carefully taken from Fish was divided into 5 groups in dissected fish and kept in cold phosphate fiberglass tanks (750 L capacity) in duplicate. buffer saline (pH 7.2) and then stored at –80°C The control group was fed a basal diet (0%) until used. and other four groups received basal diets 2.6. Determination of growth performance and incorporated with fenugreek at (1 and 2%) and biosomatic indices liquorice at (1 and 2%). All experimental After 90 days feeding, ten fish per groups were fed twice daily at 15 % of their replicate (20 fish per group) were weighted body weight. Each two weeks, fish weighted and length of each fish was measured for to adjust the feeding rate and the rate evaluating growth parameters including final nd decreased to 10% in the 2 month and to 7% body weight , weight gain (WG), weight gain rd in the 3 month of experiment. The water rate (WGR), length gain rate (LGR), feed temperature was adjusted to 25±2 °C and the conversion ratio (FCR), and feed efficiency oxygen level was maintained at 5mg/L. All ratio (FER). Moreover, specific growth rate tanks were siphoned to remove excreta and (SGR) was determined according to the two third of water was exchanged daily. Dead formula, SGR= [(Ln (Wf) _ Ln (Wi) /number fish was also recorded. of days] ×100. Where, Wf: final weight ; Wi: 2.4.2. Stress experiment Initial weight At the end of feeding experiment, two Hepato-somatic (HSI) and Spleno- sub-groups from all treated fish and control somatic (SSI) according to Yun et al., (2012), were distributed into well prepared glass and Intestino-somatic index (ISI) according to aquaria (30×40×90 cm) in duplicate (n=20 Zhang et al., (2014) were estimated. fish). The first sub-group was exposed to 2.5. Immuno-haematological parameters starvation by holding the experimental diets assay for six days. Total erythrocyte (RBCs) and leucocyte The second sub-group was subjected to number (WBCs) were counted as Kanaev, hypoxia stress by stopping the aeration with (1985). Hemoglobin concentration and measuring dissolved oxygen conc. (DO) every differential leukocytes count (DLC) were 357 Basha et al.,2018 BVMJ-34(3): 355-370 detected by Stoskopf, (1993); Packed cell differences between groups using the volume (PCV %) and blood indices (mean statistical package for the social sciences corpuscular volume (MCV), mean corpuscular (SPSS) software (Version 17.0). A value of P hemoglobin (MCH) and mean corpuscular < 0.05 was considered significant. hemoglobin concentration (MCHC) were estimated as described by Dacie and Lewis, 3.