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EFFECTS OF DIFFERENT SALINITY ON SURVIVAL RATE
OF MARBLE GOBY (Oxyeleotris marmoratus) LARVAE
001 PEY THENG
THIS DISSERTATION IS SUBMITTED IN PARTIAL FULFILLMENT OF THE
REQUIREMENTS FOR BACHELOR OF SCIENCE WITH HONOURS
AQUACULTURE PROGRAM
SCHOOL OF SCIENCE AND TECHNOLOGY
UNIVERSITI MALAYSIA SABAH
11
DECLARATION
I declare that this dissertation is the results of my own independent work, except where
otherwise stated.
March 2004
001 PEY THENG
HS2001-2243
iii
AUTHENTICATION
AUTHENTICATED BY MEMBERS OF DISSERTATION COMMITTEE
1. SUPERVISOR
(Associate Professor Dr. Shigeharu Senoo)
2. EXAMINER 1
(Mr. Kennedy Aaron Aguol)
3. EXAMINER 2
(Mrs. Rossita Shapawi)
4. DEAN
(Associate Professor Dr. Amran Ahmed)
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iv
ACKNOWLEDGEMENT
This dissertation has accumulated many debts of gratitude throughout its completion.
Thus I would like to extend my heartiest appreciation to a great number of people who
have contributed in my conquest of completing this dissertation. This dissertation would
not have been successful without their full commitment. Firstly, I would like to express
my sincere gratitude to Director of Borneo Marine Research Institute of Universiti
Malaysia Sabah, Prof Dr. Ridzwan Abdul Rahman for support in this study. Sincere
appreciation is also extended to Prof Madya Shigeharu Senoo who has been a great help
in constructive advices on my work, valuable suggestions, continuous encouragement at
various stages throughout the completion of dissertation, and providing me with a wide
range of references, also for his expert guidance and for his patience, without which this
work could not have been accomplished. I'm also greatly in debt for Mr. Kennedy Aaron
Agoul and Mr. Arun Prasad Baidya for their valuable advices, teaching and guidance. Last but not least, I would like to thank research assistants Ms. Sow Sok Hui, Ms. Nguang
Siew Ing, Mr. Ha Hou Chew and all staff of the hatchery of Borneo Marine Research
Institute, who had rendered assistance, support and cooperation in various ways-
V
ABSTRACT
Natural spawning of brood fish marble goby (Oxyeleotris marmoralus) was carried out in
960-liter square glass aquarium and 48 - 72 hours after spawning larvae were then
collected. Survivals of ten larvae in different salinities were determined at salinities from
0 to 10 (in 2 ppt increments) for 11 days in preliminary trial. Indication obtained when larvae in 10 ppt achieve greatest survival and health. Two experiments with higher
salinity were then done; from 0 to 28 ppt (0,10,20 and 28 ppt); and 0 to 25 ppt (0,10,20
and 25 ppt) in 40 days. One hundred larvae were used in 9-liter plastic aquarium with
aeration supplied. Desired salinities were increased 5 ppt within 1 hour. Water renewal,
survival rate and larval observation were conducted daily. Analyses of data revealed that
only survival in 10 ppt treatment was significant different between salinities (P<0.6 in
experiment I and P<0.3 in experiment II). No larvae could survive more than 12 days
after spawning in freshwater (0 ppt). Highest survival was achieved in 10 ppt. In 10 ppt
treatment, the greatest survival was obtained in one of the triplication with 33% of
survival rate (11% in mean) in experiment I; and 48% of survival rate (22.67% in mean) in experiment II. O. marmoratus larvae were able to survive and grow at salinities
ranging from 10 to 25 ppt. Analyses found no significant different (P>0.05) in the growth
rate for all treatments. The mortality occurred in the early larval stages 3-7 days after
spawning were much higher than those in the larval stages.
V1
KADAR KEMANI)IRIAN BAGI LARVA IKAN KETUTU
(Oxyeleotris marmoratus) DI BAWAH SALINITI YANG BERBEZA
ABSTRAK
Peneluran semulajadi bagi induk ikan ketutu (Oxyeleotris marmoralus) telah dijalankan
dalam akuarium segiempat berisipadu 960-liter dan anak ikan berumur lingkungan 48 -72 jam kemudian dikumpul. Kadar kemandirian untuk 10 anak ikan ditentukan dalam
kemasinan dari 0- 10 ppt (dengan kenaikan 2 ppt) selama 11 hari sebagai percubaan kajian. Keputusan kajian percubaan menunjukkan anak ikan dalam 10 ppt hidup paling lama dengan keadaan baik. Seterusnya 2 kajian dengan saliniti yang lebih tinggi
dijalankan iaitu 0- 28 ppt (0,10,20 dan 28 ppt); serta 0- 25 ppt (0,10,20 dan 25 ppt)
selama 40 hari. Seratus anak ikan diletakkan dalam akuarium berisipadu 9-liter dengan
kelengkapan pengudaraan. Kemasinan yang diingini dinaikkan 5 ppt dalam I jam.
Penukaran air, kadar kemandiran dan pemerhatian anak ikan dijalankan setiap hari.
Analisis data hanya menunjukkan 10 ppt mempunyai perbezaan bererti antara saliniti (P<0.6 bagi experiment I dan P<0.3 bagi experiment II). Tidak ada larva yang boleh
bertahan hidup lebih daripada 12 hari selepas menetas dalam air tawar (0 ppt). Kadar
kehidupan paling tinggi dicapai pada 10 ppt. Dalam rawatan 10 ppt, kadar kemandirian
tertinggi dicapai di salah satu daripada tiga replikasi dengan 33% kadar kehidupan
(dengan purata 11%) dalam kajian I; dan 48% kadar kemandirian (dengan purata 22.67%)
dalam kajian H. Anak ikan O. marmoratus berkebolehan hidup dan membesar dalam
lingkungan air masin 10 - 25 ppt. Analisis menunjukkan tiada perbezaan bererti (P>0.05)
dalam kadar pertumbuhan bagi semua rawatan. Kadar kematian berlaku pada peringkat 3
-7 hari selepas menetas adalah lebih tinggi daripada peringkat yang lain.
vii
LIST OF CONTENTS
DECLARATION
AUTHENTICATION
ACKNOWLEDGEMENT
ABSTRACT
ABSTRAK
LIST OF CONTENTS
LIST OF TABLES
LIST OF FIGURES
LIST OF PHOTOS
LIST OF ABBREVIATIONS
CHAPTER 1 INTRODUCTION
1.1 Malaysia Aquaculture Scenario
1.2 Marble Goby (O. marmoratus) 1.3 Objectives
CHAPTER 2 LITERATURE REVIEW
2.1 Spawning
2.2 Egg Hatching
2.3 Early Larvae Development
2.4 Mortality
2.5 Food Supply
2.6 Behavioral Changes
2.7 Salinity Effects
CHAPTER 3 MATERIALS AND METHODS
3.1 Brood Fish Selection
3.2 Hormone Treatment
3.3 Preparation of Spawning Aquarium
3.4 Fish Spawning
Page
ii
111
IV
V
vi
vii
ix
X
xi
xii
I
I
3
6
7
8
9
10
11
11
13
14
16
16
17
17
18
viii
3.5 Egg Developments and Hatching
3.6 Experimental System
3.61 Preliminary Trial
3.62 Experiment
3.7 Analysis Data
CHAPTER 4 RESULTS
4.1 Survival Rate in Preliminary Trial
4.2 Survival Rate in Experiment I
4.3 Survival Rate in Experiment II
4.4 Larvae Measurements in Experiment II
CHAPTER 5 DISCUSSION
5.1 Preliminary Trial
5.2 Comparison Survival in Experiment I and II
5.21 Freshwater -0 ppt Treatment
5.22 10 ppt Treatmant
5.23 20 ppt Treatmant
5.24 25 ppt Treatmant
5.25 28 ppt Treatmant
5.3 Peak Mortality
5.4 Feeds and Feeding Regime
5.5 Analyses Data
CHAPTER 6 CONCLUSION
REFERENCES
APPENDIX -1
19
20
20
21
23
24
24
26
29
32
33
33
34
34
36
37
37
38
38
38
39
40
41
45
ix
LIST OF TABLES
Table No. Page
4.1 Water quality during preliminary trial 24
4.2 Water quality during experiment I 26
4.3 Water quality during experiment II 29
APPENDIX -1
Table No. Page
1.1 Brood measurement, fertilization rate, hatching rate and total hatched 45
larvae in the preliminary trial and experiment 1.2 Larvae measurement for preliminary trial and experiment used 46
1.3 Comparison of changes in survival rate (preliminary trial) 47
1.4 Comparison of larvae survival for every treatment in experiment I 48
1.5 Mean survival for different salinities in experiment I 49
1.6 Analyses oneway ANOVA and Post Hoc Tests for survival rate 50
in experiment I
1.7 Comparison of larvae survival for every treatment in experiment II 52
1.8 Mean survival for different salinities in experiment 11 53
1.9 Analyses oneway ANOVA and Post Hoc Tests for survival rate 54
1.10 Comparisons growth in different salinities for experiment II at 43 d AS 56
1.11 Analyses oneway ANOVA and Post Hoc Tests for larvae 57
measurement in experiment II at 43 d AS
x
LIST OF FIGURES
Figure No. Page
1.1 Comparison of 0. marmorulus production in Malaysia from 2
year 1988-1997
4.1 Comparison of changes in survival rate (preliminary trial) 25
4.2 Mean survival for different salinities in experiment 1 28
4.3 Mean survival for different salinities in experiment 11 31
4.4 Comparisons growth in difTerent salinities for experiment 11 at 43 d AS 32
xi
LIST OF PHOTOS
Photo No. Page
1.1 Marble Goby (Oxyeleotris marmoratus) with color in alcohol dark 3
brown above and pale brown below; body with a series of large,
dark blotches; fins with dark bands or dusky
3.1 Selected brood fish were given intramuscular injection 17
3.2 Spawning of O. marmoratus 18
A. 960-liter circular aquarium with filter system B. Substrate positioned as net house inside aquarium
3.3 48-72 hours hatching larvae with slightly pigmented eyes 19
3.4 Materials used for larvae rearing in preliminary trial 20
3.5 Materials used in experiment 21
A. 9- liter plastic aquarium with green water B. Control aeration were placed aside in aquarium
3.6 Brachionus sp. (<200 gm) feed as initial food for larvae 22
3.7 Management of water quality 23
A. Bottom cleaning by using siphoned tube
B. Collected and separated larvae by beaker after siphoned 5.1 Fungus was found attacked to alive larvae 35
xii
LIST OF ABBREVIATIONS
°C degree centigrade
% percentage
BW Body weight
cm centimeter d AF days after fertilization
d AS days after spawning
DO dissolved oxygen
E East
g gram
GDP Gross Domestic Product
H hour
HCG Human chorionic gonadotropin
IU International Unit
km kilometer
km2 kilometer square
1 liter
ml milliliter
mm millimeter
mt metric ton
N North
ppt part per thousand
Sig. Significance
sp. species
SPSS Statistical Package for Social Science
TL Total length
UMS Universiti Malaysia Sabah
1
CHAPTER 1
INTRODUCTION
1.1 Malaysia Aquaculture Scenario
Malaysia is blessed with numerous natural resources. Situated in the heart of South-East
Asia, between 1 °00' N to 6°30' N latitude and between 100 °E to 120 °E longitudes; it
lies at the crossroads of the major air and sea links within the region as well as the world.
It has a total area of about 329,758 km2 and 4,400 km of shorelines and has about 23
million populations in this country. The climate is generally warm throughout the year
with daily temperatures ranging around 30 °C. The total annual rainfall averages around
200 cm. The monsoon season weather months in the West Coast are between April to
August and from October to March in the East Coast. The abundance of sunshine and
rainfall account suitable for fish to spawn which rivers and swamps are naturally covered
by forests (Ang, 1990).
The fisheries sector in Malaysia plays an important role in providing fish as a
source of food and protein (Department of Fisheries Malaysia, 2000). In 2000, it
contributed about 1.6% to the National Gross Domestic Product (GDP) and provided
2
direct employment to 81,994 fishermen and 21,774 fish culturists. During the same
period, aquaculture sector recorded a production of 167,894 tons, which constituted about
11.6% of the total fish production. However, freshwater aquaculture production was
50,688 tons, contributing 30.2% of the total aquaculture production in Malaysia.
In 1997, freshwater fishes contributed the second highest after Mollusc
production. Of this, Marble Goby (Oxyeleotris marmoratus) contributed only 39 tones on
freshwater fishes (Fig 1.1). Although the production had increased, but the production of
0. marmoratus is still very low.
Marble Goby Production in Malaysia from Year 1991-1997
1991 1992 1993 1994
Year
1995 1996 1997
Fig 1.1: Comparison of 0. marmoratus production in Malaysia from year 1991-1997
Source: Department of Fisheries, 1997
1.2 Marble Goby (0. marmoratus)
Over the last several decades, people's increasing dependence on cultured O.
marmoratus, as opposed to wild fish, as a source of dietary protein has resulted in an
increasing level of scientific effort being directed toward propagation techniques
(Donaldson et al., 1983). That are indication that fish farmers wish to culture O.
marmoratus but their seed supply is inadequate. Presently, heavy mortality in the early
stages is one of the stumbling blocs especially after the yolk sac is completely absorbed
and the larvae switch to active natural feeding. Although artificial seed production can be
achieved now, the techniques have yet to be established (Senoo el al., 1994a).
Photo 1.1: Marble Goby (Oxyeleolris marmoralus) in light dark brown above and pale brown below; body with a series of large, dark blotches; fins with dark bands
or dusky
4
Presently, the natural resource of 0. marmoratus fisheries have be exhausted in
West Malaysia (Senoo et al., 1994a). Thus, fish middlemen in the Federal capital of
Kuala Lumpur are importing this fish from Indonesia, Thailand, Cambodia, and East
Malaysia. In Sabah, wild marble goby is harvested from its natural habitat mostly in
paddy field and freshwater reservoirs. In Peninsular Malaysia, it is cultured in ponds and
former mining pools either in through monoculture or polyculture with tilapias. Cage
culture in lakes and rivers has also been successful (Cheah et al, 1994).
Although local fish farmers success culture 0. marmoratus in ponds but there are
still some attempts. These attempts, however, are on a very limited scale. This is
primarily due to inadequate supply of seed for stocking in the ponds; the seed are not
readily available in great numbers from natural sources. Thus, in order to enhance the
culture of 0. marmoratus, which could be justifiable economically, it is necessary to
increase seed availability through controlled breeding (Tan and Lam, 1973).
However, even 0. marmoratus seeds have been produced artificially, the mass
seed production has remained a major constraint. This is mainly a result of low hatching
and larval survival rate (Senoo et. al., 1994b). Early and late hatching resulted in high
mortality. The optimum hatching stage was considered just before and after the
embryonic eyes had become slightly pigmented. All embryonic heads were located at the
basal part of the egg and this was taken as normal development for 0. marmoralus.
5
Synchronistic of food production and first feeding is critical for larval survival.
Starvation may account for high mortality and the ability to endure fasting is of major
importance for survival. Heavy mortality usually occurs early in the life history of fish if
suitable food is scarce, especially when the yolk sac is exhausted and the larvae must
begin active feeding. Instead of the mouth size is suitable feed with Brachionus sp.,
however the freshwater Brachionus sp. is hard to culture under the artificial condition.
In Thailand, Tavarutmaneegul et al. (1988) successfully produced the marble
goby seeds under a semi-natural breeding and rearing method. The newly developed
larvae were first fed with a combination of chicken-egg slurry and live rotifers
(Brachionus sp. ) and then older larvae were fed with live Moina sp., chironomid larvae,
and thrash fish. However, no information is available concerning culturing technique and
the survival rate. In Malaysia, the seeds were produced in an intensive artificial rearing
method (Senoo et al., 1994a and 1994b), but the survival rate was only 10.1% during the
first 5- 70 days after fertilization (Senoo et al., 1994a).
Koumans (1953) reported the occurrence of 0. marmoratus in estuaries as well as
rivers. It may occur in brackish habitat within its range, but all of the well-documented
locality records appear to be from freshwater. Studies of the behavior and ecology of
various stages of fish development related to the causes of mortality may provide much
needed information. The aquaculture of fish depends on an adequate knowledge of the
behavior of the early stages that cause the greatest technical difficulties in culture.
6
Therefore, it is important to know the condition and behavior of 0. marmoralus larvae in
different salinity, which could improve survival rate of cultured larvae.
1.3 Objectives
In order to overcome some of the constrains of 0. marmoratus seed production,
the following objectives were outlined for this study:
1. Determination of optimum salinity for a higher survival rate of larvae.
2. Behavioral observation of larvae under different salinity.
7
CHAPTER 2
LITERATURE REVIEW
The Eleotridae or sleepers comprise some genera and 150 species of mostly tropical,
marine, brackish, and freshwater fishes. Two genera and four species occur in the fresh
waters of western Borneo. Oxyeleotris is unusual among eleotrid genera in being almost
entirely restricted to fresh water species of the genus occur in Southeast Asia and others
in the Austrian region. Three species known from western in Borneo are Oxyeleotris
urophthalmus, O. urophthalmoides and O. marmoratus (Bleeker, 1874).
Instead of freshwater eleotrid, 0. marmoratus can grows to more than 50cm in
total length (TL) and 2 kg in weight (BW). 0. marmoratus, which is also known as
Marble goby, is found in Cambodia, Fiji, Indonesia, Malaysia, the Philippines, Singapore,
Thailand and Vietnam (Cheah et al., 1994). It is much relished as a delicacy in this part
of the world where it is known as "Sand Goby" in English, "Ikan Ketutu" or "Ikan
Hantu" in Malay and "Soon-Hock" in the Chinese Fukien and Cantonese dialect, and
nowadays it is also called "Bamboo Fish" (Senoo et al., 1994a).
8
Respectively 0. marmoratus is a popular freshwater fish of high market value in
Southeast Asia. A table-sized live fish weighing lkg commands an auction price ranging
from RM60/ kg to RM80/ kg in Malaysia, prices being higher during the festive season. It
has white fine-grained sweet flesh without small bones. Consumer can get live fresh 0.
marmoratus in the market. It has become a popular dish in the Asian region from China
to Indonesia. In Malaysia, a dish of 0. marmoratus is much more expensive than other
freshwater or seawater fish, such as Sea Bass (Lates calcarifer) and Red Snappers
(Lutjanus argentimaculatus). Besides being a food fish, 0. marmoratus is also popularly
kept as an ornamental fish among the Chinese.
2.1 Spawning
The availability of a good and healthy brood fish is a pre-requisite for any successful
artificial seed production programme of fish. 0. marmoratus can mature all year round
under proper rearing conditions in Malaysia. Under good weather condition and water
temperature between 27 - 32 °C, this fish can spawn naturally in ponds or tanks. Thus, the
brood fish should be reared separately in ponds or tanks for the planned egg collection in
the seed production (Senoo et al. 1994b).
Induced fish breeding programme using hormones is increasingly being practiced
in aquaculture. Human chorionic gonadotropin (HCG) is a hormone secreted by human
placenta during pregnancy that has often been used successfully. The first success of
induced spawning of 0. marmoratus in Malaysia was reported by Senoo el al. (1992) and
9
since then, several technical improvements have been achieved (Senoo et al., 1993).
However, natural spawning is easier if compare with induced breeding under artificial
condition.
2.2 Egg Hatching
According to Senoo et al. (1994b), there are not many developmental differences in eye
pigmentation or yolk sac absorption between unhatched embryos and hatched larvae of
the same time. The differences in hatching time were not dependent on the stage of
embryonic development. In all development stages, the embryonic head was located at
the basal part of the eggs.
During hatching, larvae broke free of the egg by pushing its tail against the side of
the egg membrane. Once a suture had been made, the tail pushes through the opening.
Then, by trashing movements of the tails, the larvae freed itself from the egg, which was
anchored down by its polar threads. The larvae were unable to free themselves completely
from the eggs membranes; they swam around for sometime with the eggs membrane
covering the head and trunk like a massive hood, and died within 1-2 hours (Tan and
Lam, 1973).
10
2.3 Early Larvae Development
At the time of yolk sac absorption, Glossogobius olivaceus larvae smaller than 0.
marmoratus larvae fed on Brachionus spp., but 0. marmoratus larvae did not, in spite of
having well-developed eyes, jaws, and intestinal tracts. This might be due to the smaller
mobility of 0. marmoratus. In many marine fish larvae, the eyes, jaws, and intestinal
tracts are functional just before yolk sac absorption and larvae have small mouths, but
their locomotory organs are not well developed. Therefore, the diets for the first feeding
of larvae are limited to small-plankton which are slow in locomotion (Senoo et al.,
1994b).
In newly hatched larvae at 49 h AF, the mouth and anus were not formed, and the
eyes were not pigmented. Most of the larvae hatched during 2-3d AF; newly hatched
larvae lay on the tank bottom and gradually exhibited the "swim up, sink down" behavior.
On 3d AF, the mouth was opened, and the pectoral fins were formed. On 4d AF, the
eyes were deeply pigmented, and the lower jaw and intestinal tracts began to move. At
this time, food supply is necessary (Senoo et al., 1994b).
During seed production of 0. marmoratus, deformed juveniles were found at a
very extremely low survival rate. The seeds had a deformation at the mouth, operculum,
caudal peduncle, pelvic fin, and occipital region. The inability of the fragile larvae to
consume enough live organisms in their early stages was attributed to their morphological
deformations due to poor conditions of the brood fish or as a result of artificial spawning.
11
These deformations were considered to be disadvantageous in feeding, and eventually
resulted in poor growth and survival (Senoo et al., 1994b).
2.4 Mortality
Very high percentages (90%) of fertilization and hatching were obtained although the
larvae did not survive for more than a few days. The problem of larvae mortality has also
been encountered in the induced breeding of other fishes, for example the grey mullet,
Mugil cephalus. It is possible that the 0. marmoratus larvae died from an attack by
ciliates, which was seen to occur frequently, especially when there was a concentration of
ciliates on dead larvae nearby (Tan and Lam, 1973). Mass mortality of larval and juvenile
fish might also occur if the food supply is inadequate (Houde, 1978).
2.5 Food Supply
The food supply during larval stage is an important factor to achieve high growth and
survival rates. In different species, the sequential food in early stages differs. In the early
larval stage, the freshwater fish are generally given Brachionus sp, Anemia or Moina as
an initial diet. At the later stage the larvae or juveniles are fed minced fish, shellfish and
shrimp or on artificial pellet. It is obvious that fish of different species require different
feeding techniques (Amornsakun et al., 2003).
41
REFERENCES
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Ang, K. J., 1990. Status of Aquaculture in Malaysia. Aquaculture in Malaysia, 265-279.
Baylon, J. C., Failaman, A. N. and Vengano, E. L., 2001. Effect of salinity on survival and
metamorphosis from zoea to megalopa of the mud crab Scylla serrala forskal
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of Western Borneo (Kalimantan Barat, Indonesia). California Academy of
Sciences, San Francisco, 166-167.
Bleeker, P., 1877. Oxyeleotris marmorata. In: Inger, R. F. and Chin, P. K. (editor) The
Fresh-water Fishes of North Borneo. Natural History Publications (Borneo), Kota
Kinabalu, Malaysia, 175.
Cheah, S. H., Senoo, S., Lam, S. Y. and Ang, K. J., 1994. Aquaculture of a high-value
freshwater fish in [Malaysia] : The Marble or Sand Goby (Oxyeleotris
marmoratus, Bleeker). Aquabyte Section, 22-25.
Department of Fisheries, 1999. Annual Fisheries Statistic 1997, Volume 1. Department of
Fisheries, Malaysia, 217.
42
Donaldson, E. M. and Hunter. G. A. 1983. Induced final maturation, ovulation and
spermiation in cultured fish. In fish Physiology (Hoar, W. S., Randall, D. J. and
Danaldson, E. M., eds. ). New York : Academic Press. Vol. IX (B), 351-403.
Forsgren, E., 1997. Mate sampling in a population of Sand goby. Animal Behaviour 53,
267-276.
Huang, W. B., Chiu, T. S. and Shih, C. T., 2000. Effects of salinity of egg and early larval
characteristics of the black porgy Acanthopargus schlegeli. The Israeli Journal of Aquaculture-Bamidgeh 52 (2), 61-69.
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