The Development of Gatul Fish (Poecilia sp.

) Embryo from a Spring in State University of Malang Area

Farichah1), Dwi Listyorini2)
1, 2)

Departement of Biology Faculty of Mathematics and Natural sciences State University of Malang, Jl. Surabaya 6 Malang 65145 Indonesia

ABSTRACT
Gatul fish is widely known as guppies. According to its morphology Gatul fish is belongs to Genus Poecilia of Familia Poeciliidae. This fish bears the embryos and deliver it as larva. Gatul fish is broadly used as bioindicator for pollution and to eliminate malaria problem in many South American countries. Yet, in Indonesia the biological study upon Gatul fish is very limited. This research was aimed to study the development of Gatul fish embryo. Embryos from the ovary of gravid females from a spring in State University of Malang area were served as subject of this research. Different stages of embryos were examined to study its development. This current study showed that Gatul fish we had studied is a Livebearer. Females develop only one ovary. In that ovary ovum and embryos develop together in different stages. The division type of embryo is meroblastic, embryonic body develops at the animal pole. At the later development the body of embryo embraces the yolk sack. Yolk is absorbed into the body of newborn soon after delivery and yolk sack becomes abdominal wall. The number developing embryos in one ovary range 13 to 22. The size of embryos range from 2,8 to 3 mm in diameters or 6 mm in length from snout to tail. Keywords: Gatul fish, embryo development, Poecilia sp.

INTRODUCTION Gatul fish is a member of Family Poeciliidae. These fish live in river, pond, swamp, and lake in almost all regions in Indonesia. Gatul fish can also survive a polluted environment. Thats why it can be used as bioindicator. Beside, South American societies use this fish to eliminate the problem of malaria because this fish eat mosquito larva, but less utilized by Indonesian society. For this reason the South American countries exported Guppies tropical and sub tropical countries such as: Spain, southern France, Italy, the Balkan Peninsula, southern Romania, Ukraine, Caucasus, and Asia 1). As a member of Poecilidae Gatul fish is Livebearer. Fertilization is internal, male has external copulatory organ named gonopodium. It has a fast reproductive rate with a marked number of ovum found in the ovary which ranged from 15 to 20. The embryos develop in the ovary and delivered through cloaca 2). In our preliminary study we found that the embryos of Gatul fish developed in ovary in different stages. The number embryos ranged from 15 to 22. Based on that finding we examined the development of Gatul fish embryo from adult females living in State University of Malang area. MATERIALS AND METHODS Materials (Animal) Embryos from gravid female of Gatul fish (Poecilia sp) living in Spring in State University of Malang area.

Email: 1) farichahrozi@yahoo.com, 2) listyorini.um@malang.ac.id

Methods Gravid females of Gatul fish were captured using fishing nets and maintained in small aquarium for further examination. Captured fish then decapitated. To open the abdomen dissected the ventral site starting from cloaca to the anterior using fine scissors. Opened abdomen was examined using dissecting microscope. Ovary then isolated and placed in a petri dish containing spring water. The thin connective tissue of the ovary carefully teared to expose the embryos using fine pinsets. The embryos then examined under dissecting microscop to determine its stage. During the examination the embryos are soaked in spring water in Petri dish. The determination of embryonic development was refered to Medaka fish (Oryzias latipes) development 3).

RESULTS AND DISCUSSION

Fig. 1: Reproductive system of female Gatul fish. (A) The ovary in the abdominal cavity; circle of green dots indicate the ovary. (B & D) Position of reproduction system related to digestive system. (B-D) green bracket indicate the ovary, black arrows digestive system, blue arrows indicate the liver, orange bracket indicate oviduct, green arrows indicate cloaca, white arrows indicate ovum has vitellogenesis, red arrows showed ovum that has not yet done vitellogenesis, orange arrows indicate mesovarium , purple arrows indicate the embryo, the yellow arrows indicate ovary wrappers. (C) isolated reproductive system. Yellow line = 5 mm.

Female reproductive organs of fish Gatul composed of ovary and oviduct. There is only one ovary developed (Fig. 1A-D). The ovary is surrounded by the digestive system (Fig. 1A). This organ is bound by connective tissue (mesovarium) to other organs (Fig. 1C). The ovary might
Email: 1) farichahrozi@yahoo.com, 2) listyorini.um@malang.ac.id

contain only developing ovum or developing ovum and embryos (Fig. 1D; 2F-I; 3B & D). Ovary and oviduct leads to the cloaca (Fig 1B & C). Each ovary is wrapped with transparent thin connective tissue. In the ovary of mature female we found embryos at different stages of development as had been found in mature Poeciliopsis retropinna and Heterorandria farmosa as well 4). The number of embryos produces by Gatul fish a bit different compared to the other member of Poeciliidae in general.. Each embryo in the ovary is covered by a transparent membrane which originated from follicles sack. Ovum and embryos size varies between 2.7 mm - 3 mm. Individual females deliver its larvae developed in the ovary as of other Livebearer such as Guppy fish (Poecilia reticulata) (2). Females have the ability to store sperm, so the sperm can fertilize the developed ovum. Early adults produce 10 to 15 embryos, while the older produces 18 to 22 embryos. Member is a bit different from the number of embryos produced by other 2).

Fig. 2: The ovary of gravid female. (A) Embryo and ovum in the ovary; green arrow indicates the embryo, the red arrow indicates the egg, (B) - (I) embryos in the ovary; blue arrows indicate heart, purple arrows indicate eyes, black arrows indicate vertebra, green arrows indicate an ovum that has not yet done vitellogenesis, red arrows indicate the ovum that has done vitellogenesis, pink arrows show blood vessels, green brackets indicates the early stage of embryos.

The stages of embryonic development in different ovaries are presented in Fig. 3A-D. Fig. 3A&C shows ovaries which contain embryos in close developmental stages, while Fig. 3B&D show ovaries which contain developing both ovum and embryos. Fig. 3A,2-4 show the embryos that already have heart and eye but the body and eyes pigment have not clearly seen yet. Beside, there are more advance development which already developed eye pigment and dorsal aorta (Fig. 3A5).

Email: 1) farichahrozi@yahoo.com, 2) listyorini.um@malang.ac.id

Fig. 3: Developmental stage of the embryo from different ovaries, there is an ovary contains embryos in different of development embryos in one ovary; green arrows indicate the heart, blue arrows indicate the eyes, (A2) The body of the embryo is not yet visible. (B) - (D) embryos other ovaries; green arrows indicate an ovum that has not yet done vitellogenesis, red arrows vitellogenesis ovum, white arrows indicate vertebrae, orange the arrow indicates yolk, blue arrows indicate eyes, yellow arrows indicate brain, black arrows indicate blood vessels.

Fig 3B1-2 shows embryos that the vertebrae, pigment bodies, brain, and eyes have perfectly formed. In the one ovary there is also an ovum that has not been vitellogenesis (Fig 3B-C). Embryo has a size of a large yolk sack before birth.Yolk is absorbed into the body of the embryo (Fig. 3D) and the embryo soon became active kids swimming fish (Fig. 3D1).

Email: 1) farichahrozi@yahoo.com, 2) listyorini.um@malang.ac.id

In fig B-D there are embryos in advance development. Each embryos display a mature morphology, 1.e. head and body had been developed, eyes had become functional one with distinct pupil (black) and choroids layer (metallic). The heart has begun to beat; body pigment had been developed from head area up to tail. Soon after removal of the embryonic sack later (folicle sack) the embryos are actively swimming and its yolk immediately absorbed (Fig. 3D2). Embryonic development of Gatul fish from zygote to hatch is passing 3 stages, i.e cleavage of the embryo, development of the embryo, and free embryo respectively as refer to Peter 5). We have not yet found a complete set development stages comprising detail of each stage.

Fig. 4: The development stage of Gatul fish embryo. (A-C) early stage embryo; red arrow shows the animal pole (blastodisc). (D-Q) Developing embryos; green arrows indicate the embryo's body, green brackets indicate the body segment, white arrows indicate the embryo's body, red arrows indicate the eyes, black arrows indicate blood vessels, green arrows indicate the heart rate, purple arrows indicate heart going, Arrow indicates yolk brown, blue brackets indicate the brain, arrows showed pigment blue body, orange arrows show the vertebrae, pink arrows indicate the tail fin. (R-S) Newly hatched embryos. Green bars = 1 mm.

In this research we found a zygote (Fig. 4A), early cleavage embryos (Fig. 4B) which shows that the cleavage is meroblastic 6) occurs on animal pole as of Medaka fish (Oryzias latipes) 3) Embryo developed from those actively dividing cell at the animal pole, while yolk and yolk

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sack developed from vegetal pole. In the next stage we found the embryos that already have distinc head shape and developing eyes. At this moment this embryo has not yet formed blood vessels (Fig. 4D - E). Later, blood vessels were found in the dorsal part of the embryo's body and on the ventral side relative to the developing head (Fig. 4F). Body of the embryo's is still transparent and flattened (Fig. 4F). The next stage of development body tube was formed and became elongated; the eyes begin to form as the optic vesicle and yet there are pigment (Fig. 4G-I), dorsal blood vessels are developing (Fig. 4H); ventral blood vessel would have formed the heart and started beating (Fig. 4I-K). The optic vesicle then developed more pigment and became optic cup (Fig. 4K) and then develop into perfect eyes with metallic pigment on choroid layer, pupils is black and can be distinguished from other parts (Fig. 4L). Dorsal blood vessel and those emerged from heart the developing branched to form peripheral vessel which ventrally grow to cover yolk sack (Fig. 4M-O). Furthermore body pigment formed at the head area extends into the tail (Fig. 4N-S), eye and vertebrae almost perfect, yolk sack is shrink (Fig. 4O-S). Embryos which has reached a perfect structure hatch from follicles (embryonal sack) (Fig. 4R&S), then go out (delivered) through oviduct and cloaca. The yolk is absorbed into the body of newborn soon after delivery. This newborn is actively swimming. So far we could not yet have explanation on how those embryos are moving during the delivery. Beside, a complete set of developmental stage of Gatul fish embryo comprising the detail of each stage has not yet accomplished. However, we can show that the Gatul fish embryos development is attractive. REFERENCES
1) 2)

Hanks, H. 1971. The Pictorial Encyclopedia of Fishes. USA: The Hamlyn Pub. Eddy, S., Underhill, J.C. 1978. How to Know the Freshwater Fishes (Third edition). USA: McGraw-Hill Companies. 3) Rugh, R. 1971. A guide to Vertebrate Development (6th edition). Minneapolis: Burgess Publishing Company. 4) Bone, Q., Marshall, N.B & Blaxter, J.H.C. 1995. Biology of Fishes (Second edition). London & Oban: Blackie Academic and Professional. 5) Peter, B., Moyle & Joseph J. S. 1982. Fishes. USA: University of California. 6) Gilbert, S.F. 2006. Developmental Biology (8th Edition). USA: Sinauer Associates, Inc. Soenderland, Massacuet.

Email: 1) farichahrozi@yahoo.com, 2) listyorini.um@malang.ac.id