Journal of Agricultural Science and Technology B 2 (2012) 192-197

Earlier title: Journal of Agricultural Science and Technology, ISSN 1939-1250 D DAVID PUBLISHING

Survival and Growth of and longifilis Larvae Fed with Freshwater Zooplankton

Hyppolite Agadjihouèdé1, Antoine Chikou1, Clément Agossou Bonou2 and Philippe Adédjobi Lalèyè1 1. Laboratory of Hydrobiology and , Faculty of the Agronomic Sciences, University of Abomey-Calavi (UAC), Cotonou, Benin 2. Laboratory of Applied Biology Research, Polytechnic School of Abomey-Calavi, University of Abomey-Calavi (UAC), Cotonou, Benin

Received: July 6, 2011 / Published: February 20, 2012.

Abstract: The suitability of the freshwater zooplankton was investigated in two African larvae, Clarias gariepinus and Heterobranchus longifilis, at the end of yolk sac resorption. This live food was compared to Artemia nauplii, another live food. With those two regimens, four lots of larvae were constituted with two replicates: Cg zoo, larvae of C. gariepinus fed with freshwater zooplankton; Cg art, larvae of C. gariepinus fed with artemia; Hl zoo: larvae of H. longifilis fed with freshwater zooplankton and Hl art: larvae of H. longifilis fed with Artemia. After a 10 day-experiment, survival rates ranged from 97% to 98% and 78.5% to 81.5% were obtained in Cg zoo, Cg art, Hl zoo and Hl art, respectively. Final mean weight were 11 ± 0.003, 13 ± 0.003, 67 ± 0.001 and 89 ± 0.005 mg for Cg zoo, cg art, Hl zoo and Hl art, respectively with specific growth rate which respectively were 11.28 ± 0.3%, 14.3 ± 0.2% 14.29 ± 0.22% and 17.2 ± 0.5% per day. Those results show that freshwater zooplankton proved suitable for first feeding of C. gariepinus and H. longifilis larvae and could constitute a valuable alternative for larval rearing.

Key words: Clarias gariepinus, Heterobranchus longifilis, Artemia, freshwater zooplankton, feeding, survival and growth.

1. Introduction from endogenous to exogenous feeding, with organ formation and working bound to this function [7]. It Actually, the availability of juveniles constitutes a is considered a critical stage in many fish species in major obstacle in the development of the commercial their life history [8]. The major difficulty in larvae of African Clariid catfish such as rearing encountered lies principally in the choice of Clarias gariepinus [1-3] and Heterobranchus good-quality that was acceptable to the species longifilis [4, 5]. After mastery of reproduction of concerned. According to the literature, Artemia these species which constituted at long time main constitutes an excellent starting food in larviculture limiting factor, embryonic breeding of them remains of freshwater fish [9-12]. But due to escalation in the not mastered [6]. Now, the mastery of larviculture of coast and not availability of Artemia in our a fish species is the second indispensable link developing countries especially in the rural fish allowing to obtain its juveniles in continuous. Indeed, farming, research of substitutes which will be easily the larval period is extremely important in accessible to lesser cost is necessary. ontogenesis because it corresponds to the passage This work compares the suitability of the freshwater

Corresponding author: Hyppolite Agadjihouèdé, assistant zooplankton and the Artemia for feeding C. of research, research fields: hydrobiology and aquaculture. gariepinus and H. longifilis larvae. E-mail: [email protected].

Survival and Growth of Clarias gariepinus and Heterobranchus longifilis 193 Larvae Fed with Freshwater Zooplankton 2. Materials and Methods artificial seawater, newly hatched Artemia nauplii were separated from the hatching debris by 2.1 Source of Fish Larvae interrupting the air supply in the hatching vessel. Then, Larvae of the African catfish C. gariepinus and H. nauplii were siphoned out to a fine-mesh (100 µm) longifilis were obtained by artificial reproduction of harvesting box. The Artemia nauplii were washed and captive broodstock at the Training and Research in fed to the larvae. Fish Farming, Unit of Hydrobiology and Aquaculture 2.4 Experiment Plan Laboratory, University of Abomey-Calavi, Benin. Two females per species (around 300 g body weight Two days after post-hatching, larvae were per female) were hormonally induced to spawn using transferred in small circular baskets (Ф = 18 cm) Ovaprim (0.5 mL kg-1 body weight). The eggs directly placed in aquaria (60  30  30 cm) and harvested 10 h after, were fertiled with laitance of rearing during 10 days [15]. The initial density is 100 males (around 500 g body weights per male). Those larvae (Initial mean weight = 3 ± 0.05 mg per larvae fertile eggs were incubated in hatcheries with flowing for C. gariepinus; and 2 ± 0.01 mg per larvae for H. water using the procedure of Viveen et al. [13]. longifilis) per baskets, then 200 larvae per aquarium. Hatching occurred 27 h after incubation. After According to species and feed, four lots were hatching, the larvae were separated into the different constituted with two replicates: feeding trials with two replicates. Cg zoo: larvae of C. gariepinus feeding with freshwater zooplankton; 2.2 Mass Production of Freshwater Zooplankton Cg art: larvae of C. gariepinus feeding with artemia; Freshwater zooplankton (rotifers, cladocerans and Hl zoo: larvae of H. longifilis feeding with copepods) were mass-produced in rectangular concrete freshwater zooplankton; tanks (2  2  1 m) with manure fertilization method. Hl art: larvae of H. longifilis feeding with artemia. Each tank was half-filled using 2000 liters of water Larvae were fed in excess four times a day at 8 h to (clean water (3/5) + fishponds water (2/5) filtrates 20 h (08.00, 12.00, 16.00 and 20.00). 11th day after under plankton net of 50 µm for inoculating feeding with live food, larvae were weaned with phytoplankton) and adding chicken droppings (doses = artificial dry diet. 0.6 g L-1 attached in sack) [14]. Then the zooplankton 2.5 Water Quality and Mortality Management was inoculated. After 5-7 days of culture the freshwater zooplankton were harvested with the plankton net (55 To maintain good water quality conditions, excess µm) and thoroughly washed with tap water prior to feed and faeces were collected (siphoning) each feeding. The zooplankton used was constituted of morning before feeding; dead larvae were removed rotifers (78%), cladocerans (9%) and copepods (13%). twice daily and counted. Temperature, dissolved oxygen and pH were monitored once per day at 07.00 2.3 Production of Artemia Salina am using an Oxythermometer (DO-100 Oxygen Gauge, Artemia salina is crustacean species which leaves in Voltcraft, Hirschau, Germany; precision: 0.01 mg/L et seawater. Artemia cysts (INVE Aquaculture Nutrition, 0.1 °C) and pH meter (pH Scan 10, Oakton, Eutech High HUFA 430 µm, Hoogveld 91, B-9200 Instruments, Vernon Hills, USA), respectively. Water Dendermonde, Belgium) were incubated and hatched temperature, dissolved oxygen and pH were similar in under optimal conditions according to the all feeding lots. Mean (± SE) rearing conditions (pooled manufacturer’s protocol. 24 h after incubation in samples) were as follows: water temperature = 27.5 ±

194 Survival and Growth of Clarias gariepinus and Heterobranchus longifilis Larvae Fed with Freshwater Zooplankton

0.2 (26.1-28.8 °C); dissolved oxygen = 6.2 ± 0.1 the P < 0.05 level of significance. All statistical (6.0-6.5 mg L-1); pH = 6.7 ± 0.04 (6.2-6.7). The analysis was performed using Statview software photoperiod for this indoor experiment was set at 12 L: (Version 1992-1998, SAS Institute Inc). 12 D cycle (light period from 08.00 to 20.00 hours). 3. Results 2.6 Larvae Growth and Survival Rates Survival and growth of C. gariepinus and H. Larvae were periodically sampled to assess growth longifilis are presented in Table 1 and Fig. 1. High rates. Individual body weight was measured using an survival rates are found with Cg art (98 %) and Cg electronic balance (Sartorius, 0.001 mg and 0.01 mg zoo (97 %) with no significant difference (P > 0.05). sensitivity). Each 2 day, random and representative Hl zoo has the lowest survival. Final body weight is samples of twenty larvae were taken from each important Hl art (89 ± 0.005 mg) and Hl zoo (67 ± replicate for this measure. 0.001 mg) with statistical differences between Hl art At the beginning and the end of the experiment, and the other groups. The weak final body weights are larvae total length was measured using a millimeter obtained in Cg zoo (11 ± 0.003 mg) and Cg art (13 ± paper covered by transparent paper. 0.003 mg). For this parameter, no significant The specific growth rate (SGR), survival rate (S) difference is found between Cg zoo and Cg art. and variation coefficient (CV) was calculated using Between Cg zoo and Cg art, the SGR was statically the following formula: different. It is the same between Hl zoo and Hl art. SGR = 100 [(ln final weight – ln initial weight) / The highest SGR are obtained in Hl art (17.2 ± 0.5 % Number of experimental days] (1) day-1) and the lowest is in Cg zoo (11.28 ± 0.3 % day-1). S = (100  final number / initial number) (2) According to the mean weight gain, significant CV = (100  Standar Error / Mean) (3) differences are found between Cg zoo and Cg art and between Hl zoo and Hl art. 2.7 Data Analysis At the beginning to 8th day of larvae rearing, the Growth and survival data are presented as mean body weight curve of Cg zoo and Cg art are similar values (± SE). Results were examined by Chi care test (Fig. 2). It was the same of Hl zoo and Hl art body and analysis of variance (ANOVA 1) [16], weight curve at the first four days of rearing (Fig. 3). respectively. When ANOVA indicated significant After the 8th day at C. gariepinus and 4th day at H. treatment differences, a fisher Post Least Standard longifilis, the curves of Cg art and Hl art are Deviation test [17] was applied to compare means at respectively below of Cg zoo and Hl zoo curves.

Table 1 Survival and growth data for C. gariepinus and H. longifilis larvae feeding with freshwater zooplankton and artemia nauplii. Parameters Cg zoo Cg art Hl zoo Hl art Initial larvae number 200 200 200 200 Death number 6 4 43 37 Final larvae number 194 196 157 163 Deathly rate (%) 3a 2a 21.5b 18.5b Survival (%) 97a 98a 78.5b 81.5b Initial larvae mean weight (mg) 3 ± 0.05 3 ± 0.05 2 ± 0.01 2 ± 0.01 Final larvae mean weight (mg) 11 ± 0.003a 13 ± 0.003a 67 ±0.001b 89 ± 0.005c Mean weight gain per day (mg/day) 9 ± 0.004a 11 ± 0.007b 6.5 ± 0.003c 8 ± 0.005a Specific Growth Rate (SGR) (%/day) 11.28 ± 0.3a 14.3 ± 0.2b 14.29 ± 0.22b 17.2 ± 0.5c Values followed by the same superscript are not significantly different (P > 0.05) from others in the same line.

Survival and Growth of Clarias gariepinus and Heterobranchus longifilis 195 Larvae Fed with Freshwater Zooplankton

100 20 90 b 18 c 80 16 70 14 bb b 60 12 a 50 10 Final body weight (mg) 40 8 SGR (%/day) 30 6 20 4 a b 10 2 0 0

35 120 a 30 100 a a ab b 25 80 b 20 b 60 b

CV (%) 15 Survival (%) 40 10

20 5

0 0 Cg zoo Cg art Hl zoo Hl art Cg zoo Cg a rt Hl zoo Hl a rt Fig. 1 Effect of freshwater zooplankton and Artemia on the survival and growth of C. gariepinus and H. longifilis larvae reared during 10 days.

Fig. 2 Body weight evolution of C. gariepinus larvae feeding with freshwater zooplankton and artemia.

4. Discussion nauplii [19]. Final body weights obtained in this Under the present experiment conditions, the experiment are more important than those reported by survival and growth performances of the both species Awaïss et al. [3] on C. gariepinus larvae which are 8.7 of African catfish feeding with zooplankton approach ± 0.5. This difference could be due to live-food those obtained with artemia naupplii. The survivals feeding period which is short (7 days). H. longifilis are high and superior at 78.3%. It’s testified live food larvae feeding with zooplankton have reached more suitability on the larvae survival. This result is the important body weight in short period comparing C. same as in gudgeon (Gobio gobio) larvae [18], C. gariepinus larvae. Otémé and Gilles [16] reported gariepinus larvae fed with zooplankton (Brachionus more less body weight of same species. In 10 days H. calyciflorus) and H. longifilis larvae fed with artemia longifilis larvae fed with freshwater zooplankton

196 Survival and Growth of Clarias gariepinus and Heterobranchus longifilis Larvae Fed with Freshwater Zooplankton

Fig. 3 Body weight evolution of H. longifilis larvae feeding with freshwater zooplankton and artemia. has 67 ± 0.001 mg (mean body weight). According to encountered lies in its good digestibility. Live-foods Otémé and Gilles [2], this body weight obtained in can contribute to the own digestion until 80% due to present experiment with freshwater zooplankton is the proteases which they contain. The digestibility of sufficient for weaning H. longifilis larvae. In spite of rotifers proteins by Cyprinidae larvae ranged from initial body weight of H. longifilis larvae is 2 ± 0.01 89% to 94%, while for artemia it’s situated between mg versus 3 ± 0.05 mg for C. gariepinus larvae; final 83% and 89%. Nevertheless, African catfish larvae body weight of H. longifilis larvae is largely superior such as C. gariepinus larvae use better the proteins of of the one of C. gariepinus larvae (Table 1). What the artemia than those of the rotifers; what could shows that H. longifilis has more quickly growth than justify the difference obtained at the level of the C. gariepinus [20, 21]. Another reason which performances of survival and growth. contributed at this difference may be the mortality of 5. Conclusion larvae. Indeed, the mortality of C. gariepinus larvae was significantly lower than H. longifilis larvae. Thus, At the end of present experiment, it’s clear that C. C. gariepinus larvae had a smaller space per gariepinus larvae and H. longifilis larvae fed with individual than H. longifilis larvae. It is well known freshwater zooplankton approach the survival and that fish density is related to fish growth. growth performances of larvae of same species fed The growth rate obtained here at both species is with artemia nauplii. Then, zooplankton could similar to those obtained in C. gariepinus, Clarias substitute artemia in larviculture of African catfish. macrocephalus and Labeo parvus when comparing But, the research on the species of zooplankton, the larvae fed a dry diets and live foods (Artemia nauplii nutritional composition of zooplankton specie and the and freshwater zooplankton). rationment are necessary in to improve the The good survival and growth performances of efficiency of freshwater zooplankton. those larvae fed with freshwater zooplankton and Acknowledgments artemia nauplii obtained in the present experiment results in digestibility and of good nutrient quality of The authors thank Celia S. Fiogbé, Edwige S. both prey. According to Lauff and Hofer [22], a Houéhou and Karen Savi for their contributions to this principal interest of live-foods in larviculture work, particularly with data collection.

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