Effect of Feeding with Different Dietary Protein Levels and Starvation onthe Health, Nonspecific Immune Parameters, Behavior andHistoarchitectures of Fantail Goldfish (Carassius auratus L.)ALshimaa AHM Khalil1, Walaa EL-Husseiny1, Azhar F Abdel Fattah2 and Wael AM Ghonimi3*1Department of Fish Diseases and Management, Faculty of Veterinary Medicine, Zagazig University, 44519 Zagazig, Egypt2Department of Veterinary public health, Faculty of Veterinary Medicine, Zagazig University, 44519 Zagazig, Egypt3Department of Histology and Cytology, Faculty of Veterinary Medicine, Zagazig University, 44519 Zagazig, Egypt*Corresponding author: Wael AM Ghonimi, Department of Histology and Cytology, Faculty of Veterinary Medicine, Zagazig University, 44519 Zagazig, Egypt, Tel:00201222498246; Fax: +2-055-2283683; E-mail: [email protected]
comRec date: Sep 02, 2015; Acc date: Dec 02, 2015; Pub date: Dec 04, 2015Copyright: © 2015 Khalil AA, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse, distribution, and reproduction in any medium, provided the original author and source are credited.AbstractThe present investigation was conducted to assess the effect of different dietary protein percentages andstarvation on the health, behavior, blood chemistry, immune response and histoarchitectures of fan tail gold fishCarassius auratus L.
Experiment was carried out using 80 Carassius auratus with an average body weight 18 ± 2 gwhich were divided into four groups in duplicate in which, group 1 feed with diet A; 28% Crude Protein (CP). Fish ingroup 2 was fed on diet B (17% CP) and fish in group 3 was feed on diet C (45% CP) and group 4 kept as fastedgroup. The experimental period was 8 weeks. The current study clarified that significant high growth rate, improvedwelfare; decrease aggressive behavior, improved biochemical serum parameters and immune response wereobserved in group feed with diet containing 45% crude protein. Starvation is directly affected health, immunity andwelfare as well as histoarchitectures of all selected organs. Histologically, there is no any significant changes on thehistoarchitectures of the all selected organs; liver, spleen, intestine, head kidney as well as muscle of groups 1, 2and 3 feed with diet A (28% CP), diet B (17% CP) and diet C (45% CP) respectively. Meanwhile, the fasting of group4 had the most effective changes on the histoarchitectures of all selected organs.Keywords: Dietary protein; Starvation; Health behavior;Histoarchitectures; Fantail goldfish; Carassius auratus LIntroductionOrnamental fish have growing importance at the present time.Cichlids, which are considered the most popular ornamental fish,constitute approximately 95% of all 4000 species and varieties .There are different factors that affect ornamental fish welfare,including for example the physical and chemical environment, feeding,social interaction and the occurrence of fish pathogens . Nutrientshave an important role in keeping the health condition, normalbehavior and in improvement the external appearance and color ofornamental fish . Protein is considered the biggest part of the cost ofthe unit of feed, while fat and carbohydrates are important to supplythe energy required by fish .The changes in the metabolic and immunological profiles are one ofthe tools that evaluate the fish performance and its ability to withstandthe different dietary conditions [4-6].Histological analysis of the digestive system is considered a goodindicator of the nutritional status of fish [7-9]. The intestine and liverare the most important organs in digestion and absorption of nutrientsfrom food, and therefore monitoring of these organs is considerednecessary .Starvation is one of the important causative of fish mortality innature and in aquaculture . Long-term starvation can cause severedeformity in vital organs  and even mortality of fish. Starvationalso exhibits a widespread histological degeneration in thehaemopoietic organs of fish which can bring about alterations in theircellular architecture. Haemopoietic organs (liver, head kidney andspleen) have been reported to be the most sensitive tissues to beaffected by starvation . Structural alterations/degeneration ofhaemopoietic organs impairs its functional capacity (haemopoiesis)which may prove even fatal for the survival of fish. Thushistopathological alterations in haemopoietic organs can be utilized astools in order to get a clear idea about the extent an organism isaffected at tissue or cellular level .The aim of our work is to investigate the effect of different dietaryprotein levels and starvation on fish growth performance, health,behavior as well as histoarchitectures of liver, spleen, intestines, headkidney and muscle of fantail goldfish (Carassius auratus ).Material and MethodsFish and aquariaA total number of 80 fantail goldfish (Carassius auratus L.) with anaverage body weight 18 ± 2 g were collected alive from Zagazig fishmarket at Sharkia province and transported alive immediately to thelaboratory in large plastic bag. Fish were kept in experimental aquariawith 60 liter capacity and kept for 10 days acclimatization periodbefore the beginning of the experiment. Water in the aquaria wasaerated permanently and the temperature was regulated by usingthermostatically controlled heaters. Water was completely changedfour times weekly.Veterinary Science & Technology Khalil, et al., J Veterinar Sci Technol 2016, 7:1 Article Open AccessJ Veterinar Sci TechnolISSN:2157-7579 JVST, an open access journalVolume 7 Issue 1 1000278Fish diets and feedingDiets with different protein levels were prepared in fish researchunit, Faculty of Veterinary Medicine, Zagazige University, Egypt. Thechemical analysis of feed stuffs used in the experimental diets is shownin Table 1. The diets included different crude protein levels asfollowing: Diet (A) in which, contains 28% crude protein; diet B,containing 17% crude protein, diet C, containing 45% crude proteinand group D acts as a fasting group. The fish in groups A, B, C were feddiets 2 times daily (09:00 AM and 03:00 PM) at rate of 3% of bodyweight for 8 weeks. Periodical evaluation of growth was done each 15days. The chemical composition of the experimental diets are shown inTable 2. The growth Performance Parameters carried out by evaluationof average body weight which calculated by dividing the total weight offish by the number of fish in each group. Body gain, body gain percent,specific growth rate % and daily gain rate were determined accordingto Pouomonge and Ombredane .Ingredient Nutrient (% as fed basis)DM CP EE CF Ash NFE(calculated)Yellow corn 88.80 8.75 3.60 2.10 1.20 73.15Wheat flour 89.00 12.80 2.50 1.60 1.60 70.50Soybean meal 90.00 43.70 1.80 6.10 6.50 31.90Fish meal 94.60 63.40 8.70 0.7 20.50 1.30Poultry by-productmeal92.60 60.30 12.70 2.10 14.70 2.80Table 1: Chemical analysis of feed stuffs used in the experimental diets.(DM= Dry matter, CP=Crude protein, EE=Ether extract, CF=Crudefiber and NFE (calculated)=Nitrogen free extract). DM, CP, EE andAsh were chemically analysed according to procedures of AOAC(2000).*Calculated according to tables of NRC (1993).Ingredients Experimental dietsCP % in dietsA B CYellow corn 39.00 50.00 14.00Wheat flour 12.00 23.00 6.00Soybean meal 16.00 7.00 22.00Fish meal 13.00 6.00 27.00Poultry by-product meal 13.00 5.00 27.00Vegetable oil 5.50 7.50 2.50Vitamins and Minerals mixture* 1.50 1.50 1.50DM, % 84.14 81.57 88.19CP, % 28.02 17.20 45.01EE, % 10.04 10.89 9.16CF, % 2.38 2.02 2.50Ash, % 6.30 3.40 11.25NFE, % 42.63 55.24 22.60DE, Kcal/ kg diet** 2919.40 2915.49 2944.55Table 2: Chemical composition of the experimental diets. *Vitamin andMineral mixture (alfakema):- Each 1 kg contains:-Vit. A 580000 I.U,vit.D3 8600 I.U, vit.E. 720 mg, vit. K3 142 mg, vit C 0.1 mg, vit B1 58mg, vit B2 34 mg, vit. B6 34 mg , vit.B12 58 mg , Folic acid 86 mg,Pantothenic acid 8 mg , Manganese sulfate 65 mg , Zinc methionine3000 mg , Iron sulfate 2000 mg , Copper sulfate 3400 mg , Cobaltsulfate 572 mg , Sodium selenite 25 mg, Calcium iodide 25 mg,Calcium carbonate (Carrier substance) till 1000 gm. **Digestibleenergy calculation based on values of protein 3.5 kcal/gm, fat 8.1kcal/gm, NFE 2.5 kcal/gm (Santiago et al. 1982). (DM=Dry matter,CP=Crude protein, EE=Ether extract, CF=Crude fiber andNFE=Nitrogen free extract).Experimental designFish are divided into four groups in duplicate in which, group 1 feedwith diet A. Fish in group 2 was fed on diet B and fish in group 3 wasfed on diet C. while fish in group 4 kept as fasting group. Theexperimental period was 8 weeks.Behavioral observationCarassius auratus identified by short plastic strips applied in dorsalfin of fish, behavior recorded in the period between 09:00 Am till 03:00Pm for 8 weeks by using focal sample technique for 15 sec. intervalsduring one hour daily. Visually by using a note book for recordingbehavior, a stop watch, multipurpose counter and video cameraaccording to Altuman . The following behaviors were recordedaccording to Stephan :Feeding: Frequency and duration (Sec.) spent in feeding.Swimming: Frequency and duration (Sec.) spent in swimming.Aggression: Frequency and duration (Sec.) spent in attacking eachother.Rest: Frequency and duration (Sec.) in which fish completelyimmobile and rest on the bottom of their aquaria.Arousal: Frequency and duration (Sec.) in which fish has alocomotors activity.Fish coming to surface of aquaria: Frequency and duration (Sec.) inwhich fish hanging around the top of aquaria.Blood sampleAt the end of the experimental periods, blood samples werecollected from caudal vessels and centrifuged at 3000 rpm for 10minutes for obtaining serum samples and stored in freezer at -0°C thenanalyzed 24 hrs post collection. Blood samples should be collected 17 hafter the final feeding for the plasma glucose assay according to Chenget al. .Biochemical analysisBiochemical analysis for glucose which was determinedcolorimetrically according to Trinder , cholesterol , totalprotein  and creatinine  were determined.Citation: Khalil AAHM, Husseiny WE, Fattah AFA, Ghonimi WAM (2016) Effect of Feeding with Different Dietary Protein Levels and Starvation onthe Health, Nonspecific Immune Parameters, Behavior and Histoarchitectures of Fantail Goldfish (Carassius auratus L.). J Veterinar SciTechnol 7: 278. doi:10.4172/2157-7579.1000278Page 2 of 12J Veterinar Sci TechnolISSN:2157-7579 JVST, an open access journalVolume 7 Issue 1 1000278Immunological assessmentImmunological response of fish was evaluated throughdetermination of serum lysozyme levels  and IgM .Statistical analysisData were collected, organized and analyzed using one-way analysisof variance (ANOVA) through the general linear models (GLM)procedure of the Statistical Package for Social Sciences version 21.0(SPSS for Windows 21.0, Inc., Chicago, IL, USA). The comparison ofmeans was carried out with Duncan’s multiple range tests (DMRT).Results were recorded as mean ± standard deviation (SD). The value ofP