NAME : DEYVANAI ARUMUGAM MATRIIX: JP/5523/08 NAME OF SUPERVISOR: PROFFESOR DR.

IBRAHIM JAAFAR

INTRODUCTION

Fejervarya limnocharis ( Gravenhorst,1829) or frog of rice paddies, inhabits in open country, clearings, agricultural land, road ditches, inner city parks and other habitats created or disturbed by humans. In the field it can be identified by the long toes on its hind legs, the intermittent raised skin ridges on its ventral surface, its white belly and its relatively small size. A vertebral stripe may or may not be present. It ranges from India and Sri Lanka, through Thailand and southern China to Japan and Taiwan, and down through Peninsular Malaysia, Singapore and the major Indonesian islands

Scientific name : Fejervarya limnocharis (Gravenhorst,1829) Common name : Common Grass Frog Duration of metamorphosis : 8 -10 weeks (Etkin et al.1968) The total length : 50-60 mm (Berry, 1975) Feeding habit : Food from plants and microorganisms found within the natural environment of their habitats. (Berry, 1965)

T HE IMPORTANCE OF
AMPHIBIAN STUDY

To use as source of food and protein for human (Ibrahim,1954) To create a balance ecosystem (Ibrahim,1954) To identify the reason of frog extinction around the world. This is due to the number of frog is declining nearly 168 species are believed to have gone extinct and at least 2,469 (43%) more have populations that are declining. (Stuart et al. 2004). To learn about ecological and evolutionary pattern of amphibians To understand the survival range of amphibians. To identify use of amphibians in conservation and human health. (Ibrahim,1954)

OBJECTIVE

To study the growth and development rate of Fejervarya limnocharis (Gravenhorst,1829) larvae.
To determine the survival rate of Fejevarya limnocharis (Gravenhorst,1829) larvae in lab conditions.

To find out the duration of Fejervarya limnocharis (Gravenhorst,1829) larvae growth to metamorphosis in lab conditions.
To identify the Fejervarya limnocharis (Gravenhorst.1829) larvaes metamorphosis stages by comparing with Gosner stages,1960.

A BOUT

GOSNER S STAGES

The description of anuran embryos and larvae is facilitated by the use of staging tables. The tables are indispensible to many studies involving live history of frog. Gosners proposed table is an extension and simplified of those already in exist. Gosners stages had simplified anuran embryo and larvae metamorphosis into 46xxv stages. At the end of Gosner stage 46xxv the metamorphosis of tadpole larvae will complete.
(Gosner,1960)

G OSNER
(a)

STAGES

Embryonic stage 1- stage 25 : Contain the embryonic series

(b)
(c) (d) (e) (f) (g) (h) (i) (j)

Stage 1 : Rotation of the embryo until the animal pole is uppermost

Stage 2 :Second polar body is expelled Stage 3:stages 5- Cell division Stage 6: Cell division less regular Stages 7,8,and 9: Differentiated by the size of the blastomers Stage 8 ,9 :Light hemisphere reduction Stage 10 :Beginning of gastrulation Stage 11-12 :Blastophore formation Stage 13 : Develops as tabular area on the dorsal surface

Stage 14: Elongation of the embryo and the elevation of two lateral ridges separated by neural groove.
Stage 15: Ciliary rotation of the embryo will active Stage 16: Forming of neural tube Stage 17:Development of tail bud Stage 18,19 and 20: Differentiation basis on the relative development of the external gills and tail. Stage 21 and 23: Full development of external gills and free swimming. Stage 23,24 and 24 : Development of operculum and disappearance of external gills, differentiation of oral disc and labial tooth. Stage 25: Presence of spiracle and starting of indi Stage 26&30: Development of hind limbs.

Stage 31: Differentiation of foot into paddle shape Stage 32-37 : The development and appearance of individual toes Stage 38-40 : Appearance of metatarsal and subarticular tubercles. Stage 40 : Drastic changes of metamorphosis Stage 41: Forelimbs skin become transparent

Stage 42-46 : Appearance of forelimbs

Stage 46: Metamorphosis complete. Stage 29-40 : Mouth parts unchanged

Stage 32: Pigmentary pattern become stabilized

M ATERIAL
(a) (b) (c) (d) (e) (f) (g) (h) (i) (j)

10 plastic aquaria (2.5 liter) 1500 ml dechlorinated water 1:150ml (each aquaria)

100 tadpoles (10 tadpoles in each sample)

Small net Petri dish Lactuva sativa (lettuce) Fish flakes-media aqua fish, made in Japan Temperature recorder Graph paper (for measurement) Camera (for recording purpose)

M ETHODS
(a)

A field trip to Byram estate, Nibong tebal for eggs collection on 30.11.2011, 5.30am.
The eggs were found in water puddles.

(b)

(c)

By using scoop, the eggs were transferred into aquarium and brought to lab.
The eggs were kept for 7 days in lab condition.

(d)

(e)

After seven days the eggs which are kept in lab conditions developed into tadpoles.
The tadpoles were in Gosner stage 20.

(f)

(g) Each aquarium filled with 1500ml of dechlorinated water. Ratio 1:150ml to prevent space competition among larvae. (must use dechlorinated water to avoid mortality in tadpoles)
(h) 100 tadpoles in Gosner stage 20 were transferred into 10 aquaria . Now we have 10 tadpoles in each aquarium. (i) All the aquaria were labeled from sample 1 to sample 10. The amount of water and surrounding temperature were labeled in each sample. (j) The sample 1 to 10 were placed in laboratory under temperature range between 25C-27C (k) The development and survival rate of tadpoles were observed and recorded in chart. (l) The development stage were compared with Gosners stages. (m) Each identified stages were recorded as Gosner stage 1-30

(n) As tadpoles reached Gosner stage 21 the total length were recorded ( head to tail).
(o) Two big sizes tadpoles and two small sizes tadpoles were choose. (p) Four tadpoles from each sample were taken out and placed in Petri dish. (q) The Petri dish with four tadpoles were placed on a graph paper. (r) Observer need to wait till the tadpoles settle on bottom of Petri dish. (s) After tadpoles settled the total length were measured and recorded in a chart. (t) This step will repeat until tadpoles reach Gosner stage 46xxv. (u) Each measurement will use for further research.

The survival rate of tadpoles Observation record Stages identification using Gosner table (1960)
Day 1 starts from the day, tadpoles(GS 20) transferred into ten samples.
Days 1 3 5 7 9 11 13 15 17 19 21 Survival Rate (%) 100 100 100 100 100 79 79 79 79 69 69 Gosner stage Days Survival rate (%) Gosner stage

GS 20
GS 20 GS 20 GS 21 GS 21 GS 21 GS 22 GS 22 GS 23 GS 23 GS 24 GS 24 GS 25 GS 25 GS 25 GS 26

32
34 36 38 40 42 44 46 48 50 52 54 (31.1.2012)

52
52 51 47 46 46 46 45 45 45 45 45

GS 26
GS 26 GS 27 GS 27 GS 27 GS 28 GS 28 GS 28 GS 28 GS 29 GS 29 GS 29

23
25 27

61
61 61

28
30

60
59

*Gosner stage identified by observe the biggest tadpoles in each sample

The growth rate of tadpoles Observation record

.
Number of tadpoles
Days 25 (01.01.2012) Big Small 28 (04.01.2012) Big Small 31 (06.01.2012) Big Small 34 (09.01.2012) Big Small 37 ( 12.01.2012) Big Small 40 (16.01.2012) Big Small

Length (mean) Head to tail

19.9mm 9.3mm 20.1mm 9.6mm 21.9mm 11.95mm 23.9mm 13.5mm 25.0mm 14.5mm 25.7mm 15.5mm

Standard deviation

2.19 1.97 2.33 2.11 3.10 2.45 3.21 2.74 3.67 2.88 3.87 3.01

G OSNER

STAGES

FEEDING HABIT

E XPECTED RESULT

The growth and development of tadpoles still need another 3-4 weeks.
After 3-4 weeks the tadpoles will complete its metamorphosis cycle.

The research and observation will continue until tadpoles reach complete metamorphosis (GS 46 xxv).
Obtained data and result will use for final conclusion.

Thank you

R EFFERENCE

Berry, P. Y. (1965). The diet of some Singapore anura (Amphibia). Proc.2001. soc. London,144: 163-174. Berry, P. Y. (1975). The Amphibian Fauna of Peninsular Malaysia. Tropical Press, Kuala Lumpur. Callery, E. M. & Elinson, R. P. (2000). Opercular development and ontogenic re-organization in a diet developing frog. Dev Genes Evol, 210: 377-381. Etkin,W. & Gilbert,I.L. (1968). Metamorphosis: A Problem in Development Biology. New York. Fabrezi, M. (2011). Heterochrony in Growth and Developmemt in Anurans from the Chaco of South America. Evol Biol, 38: 390-411. Gosner, K. L. (1960). A simplified table for staging anuran embryos and larvae with notes in identification. Herpetologica 16: 183-190.

Ibrahim, J. (2004). Aspects of Biology and Ecology of Two Sympatric Frogs Species in Malaysia. Kedah:Yusran Publishing House.
Ibrahim, J. (2010). Biodiversiti Amfibia di Semenanjung Malaysia Warisan Alamiah yang Amat Berharga. Pulau Pinang: Universiti Sains Malaysia. Rollins-Smith, L.A. & Cohen, N.(1996). Metamorphosis: an immunologically unique period in the life cycle of the frog. Academia Press, San diego,Califf, 626-646. Stuart, S. N., Chanson, J. S. & Cox, N. A.(2004). Status and trend of amphibians declines and extinctions worldwide. Science 306: 1783-86. Saidapur, S. K. (2001). Behavioral ecology of anurans tadpoles : The Indian Scenario. Proc.Inidan Natri Sci Acad.(PINSA) 6: 311-322. Williamson, I. & Bull, C. M. (1989). Life history variation in a population of the frog Ranidella signifera: Egg size and early development. Copeia, 349-356.