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Embryonic and Larval Development of Rohu-Mrigal Hybrid"

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Embryonic and Larval Development of Rohu-Mrigal Hybrid Proc. Indian Acad. Sci. (Anlm. Sci.), Vol. 90, Number 4, July 1981, pp. 417-426. © Printed in India. Embryonic and larval development of rohu-mrigal hybrid" Y BASAVARAJU and T J VARGHESE University of Agricultural Sciences, College of Fisheries, Mangalore 575002, India MS received 9 October 1980; revised 13 April 1981 Abstract. A detailed study of the embryonic and larval development of the major carp hybrid, rohu-mrigal, has been made and compared with the parents. The development of the hybrid was normal and more or less similar to that of the parental species, with only a few minor differences. The hybrid larvae exhibited intermediate range between the parental species in morphological features, such as nature of lips, presence of barbels and the shape of the caudal spot. Keywords. Development; rohu-mrigal hybrid. 1. Introduction Hybridisation as a tool for improving natural stocks of plants and animals has been successfully applied to produce high yielding varieties in agriculture and animal husbandry. But its application in aquaculture is relatively a recent inno­ vation. After the development of the technique of hypophysation, a number of carp hybrids have been produced in India (Chaudhuri 1959, 1973; Varghese and Sukumaran 1971; Naseem Hamsa 1971; Naseem Hamsa and Alikunhi 1971; Konda Reddy 1977; Varghese and Shantaram 1979). But detailed evaluation of these hybrids from aquaculture point of view has been done only in a few cases. Therefore, a detailed study in development, taxonomic characters and growth of two major carp hybrids, rohu-rnrigal and mrigal-rohu, was taken up at the College of Fisheries, Mangslore. The present account deals with the embryonic and larval development of the hybrid, rohu-rnrigal, 2. Materials and methods The hybrid, rohu-mrigal was produced by crossing male rohu (Labeo rohita) with female mrigal (Cirrhinus mrigala) employing the. technique of hypophysation. The method of dry stripping was followed. Wh12n the injected female mrigal was found freely oozing, the eggs were stripped into a clean enamel tray. Immediately '" Formed part of the M.F.Sc. Thesis submitted by the senior author to the University of Agricultural Sciences, Bangalore, 417 418 Y Basavaraju and T J Varghese after this, male rohu was taken out and the milt was pressed on to the eggs. The sex products were then mixed thoroughly by gentle shaking of the tray. Excess of milt and slime were removed after a coupl of minutes by repeated washing of eggs with pond water. The fertilized eggs were kept in basins with sufficient water for water hardening and development. Following the same technique pure breeds of rohu and mrigal were also produced. Developing eggs of all the three breeds were transferred to separate hatching hapas, tied in cement cisterns, with appro­ priate labels. Before that, a sample of developing eggs were brought to the labo­ ratory for studying the development. The hatchlings, after yolk-sac absorption, were transferred to well-prepared nurseries. Samples from these were used for subsequent observations. The eggs were observed under a light source binocular microscope of 3·2 x 5 and 3·2 x 10 magnifications.. The observations were continuous from the time of fertilization till the completion of segmentation and thereafter at 30 min intervals till hatching. Mter hatching, observations were made at 6, 12, 24, 48 and 72 hr. Post-larval development was studied at intervals of 24 hr till the 7th day and thereafter on 10th, 12th, 15th, 20th and 25th days. For this, specimens freshly preserved in 4% formalin were used. Camera-lucida drawings were made under a magnification of 3·2 x 5 till the 7th day and thereafter, using 5 x or 10 x magnifications. The specimens were, however observed under higher magnifica­ tions, whenever necessary to study minute details. The developmental details of the rohu-mrigal hybrid only are described in this paper as these details of rohu and mrigal have already been published by Chakrabarty and Murty (1972). Table 1. Embryonic development of rohu-mrigal hybrid Range of water temperature: 26'0 to 30° c. Hours after Progress in embryonic development fertilization 00·40 Blastodisc formation 00·45 2-celled stage (first cleavage) 00·50 <kelled stage 00·57 8-celled stage 01'05 16-celled stage 01-45 Morula stage 03·30 Half, yolk invasion completed 05·55 Yolk-plug stage 08'10 Differentiation of embryo into cephalic and caudal ends 08-25 Comma stage, no myotomes 09-35 Appearance of optic cup; embryo with 8 mrotomes 10·30 Appearance of Kuptfer's vesicle; embryo with 16 myotomes; elongation of yolk mass 11-30 Disappearance of Kuptfer's vesicle; embryo shows slight movements 13'10 Vigorous twitching movement; 2 otoliths in-otic vesicle 16·55 Hatching started s *4_ Development of rohu-mrigal hybrid 419 3. Results 3 .1. Embryonic development The fully swollen eggs measured between 4·25 and 5·31 mm in diameter, with an average of 4·83 mm. The details of embryonic development of the hybrid are presented in table 1 and figure l(a) to (m). 3 .2. Larval development (figure 2a) The just hatched larva is transparent and devoid of mouth. Eyes are pigmentless, The yolk-sac has two portions, anterior short bulbous portion and posterior elongated portion. Myotomes, 27 + 10. Total length 4·20 mm. 6 hr old hatchling (figure 2b) The constriction between the bulbous portion and the elongated portion of the yolk-sac disappears. Two auditory concretions are clearly seen, the posterior one being slightly larger. Total length 4·44 mm. 12 hr old hatchling (figure 2c) Eyes slightly pigmented at the centre. The bulbous portion of yolk-sac is reduced in height. Myotome.j Z? + 14. Pectoral fin bud appears. Notochord is seen as a spotted line. Total length 4·82 mm. 24 hr old hatchling (figure 2d) Eyes become more pigmented, particularly at the centre. Mouth appears as a slit. Yolk-sac gets elongated. A reddish hue is seen near the opercular region. Total length 5'11 mm, 48 hr old hatchling(figure 2e) Mouth opens. Air-bladder is seen. Pectoral fin is more prominent. Intestine is visible. Gill arches are seen as lines. Ventral fin-fold starts from about the middle region of air-bladder. Dorsal fin-fold starts immediately after the air­ bladder. Caudal is nearly round with five striations. A few chromatophores appear on the head. Total length 5·44 mm. 72 hr old hatchling (figure 2f) Yolk-sac almost completely absorbed. Mouth well-developed. Number of chromatophores on the head region increases. Operculum appears covering the gill-arches. Air-bladder elliptical in shape. Notochord clearly seen with the posterior end slightly upturned. Eight striations are present on the caudal fin. A few chromatophores are seen around the tip of the notochord, Myotomes, 27 + 14. Total len~h 5· 77llUll. 420 Y Basavaraju and T J Varghese a b c Imm d e f 9 J k .I -'"'' m Figure 1. Embryonic development of rohu-mrigal hybrid. Fertilized egg­ (a) blastodisc formation, (b) 2-celJed stage, (c) 4-celJed stage, (d) 8-celled stage, (e) 16-celJed stage, (f) morula stage, (g) half yolk invasion, (h) yolk-plug stage, Embryo, (i) elongation of yolk mass, (j) comma stage without somites, (k) appear­ ance of optic cups, (1) appearance of Kupffer's vesicle, (m) embryo before hatching. Development of rohu-mrigal hybrid 421 L. hnm ... hun Figure 2. Pro-larval development of rohu-mrigal hybrid. Pro-larvae (a) just hatched, (b) 6 hr after hatching, (c) 12 hr after hatching, (d) 24 hr after hatching, (e) 48 lir after hatching, (0 72 hr after hatching. 3.3. Post-larval development 4th day after hatching (figure 30) Lips well-developed, the lower one being thicker and slightly shorter. Ventral fin-fold originates from about 3/4th the length of the air-bladder from the anterior end. Dorsal fin fold starts a little behind the air bladder. No constriction is seen in the air-bladder. Total length 6 mID, 422 Y Basavaraju and T J Varghese (gl20lhday aftel: hatching,(hl2Sth .day after hatching. Figure 3. Post-larval development of rohu-mrigal hybrid,. Post-larvae-(a) 4th day after hatching, (b) 5th day after hatching, (c) 6th day after hatching, (d) 7th day after hatching, (e) 10th day after hatching, (f) 15th day after hatching. ~) 20th day after hatching, (h) 25th day after hatching, Development of rohu-mrigal hybrid 423 5th day after hatching (figure 3b) The upper jaw is slightly longer than the lower jaw. Differentiation of dorsal fin has started, two pigmented striations appear. A small patch of dark pigments is seen below the tip of the notochord. Caudal with 12 rays, three rays on either side being unbranched. A small notch appears on the posterior margin of the caudal, indicating the commencement of bifurcation. Total length 6·55 mm. 6th day after hatching (figure 3c) Pigmentation has increased on the body and head. Air bladder is covered by chromatophores, Dorsal fin with six striations. Dorsal fin fold still persists. Anal, separated from the ventra! fin-fold, no pigmentation seen. Caudal with 16 rays. Fresh fry looks more yellow-golden on the anterio-ventral side, while it is pale-yellow on the abdominal region. Total length 6·74 mm. 7th day after hatching (figure 3d) A constriction in the air-bladder appears resulting in an anterior globular part and a posterior elongated part. Ventral fin-bud appears. Dorsal fin is separated from the fin-fold, with eight slightly branched rays. Upper and lower jaws are more or less equal in length. Tip of the notochord clearly upturned. Caudal fork is marc clear, the lower lobe being slightly longer.
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