IJERD – International Journal of Environmental and Rural Development (2014) 5-2

Research article
erd

Acute Toxicity of Tea Seed Cake on Climbing perch(Anabas testudineus (Bloch))
KITTIMA VANICHKUL*
Faculty of Agricultural Technology, Rajamangala University of Technology Thanyaburi,
Pathum Thani, Thailand
Email: kittima.va@gmail.com, kittima_v@exchange.rmutt.ac.th

VISITSAK HAMTHANON
Faculty of Agricultural Technology, Rajamangala University of Technology Thanyaburi,
Pathum Thani, Thailand

THAWORN SIRIPONG
Faculty of Agricultural Technology, Rajamangala University of Technology Thanyaburi,
Pathum Thani, Thailand
Received 15 November 2013Accepted 21 September 2014(*Corresponding Author)

Abstract Tea seed cake is common used in fish and shrimp farms in Thailand forcontrolling pond predators such as fish, shellfish or tadpole due to its applicable andaffordable price. Climbing perch (Anabas testudineus (Bloch)) is one of several problems
in aquaculture because it can lead to threatening diseases, the displacement of nativespecies, causing changes to the water quality. Moreover they may effect as destroy ordecrease of the productivity. So, acute toxicity of tea seed cake on climbing perch wasstudied. Statistic bioassay method (48-hr LC50) was a technique to determine the medianlethal concentration of the tea seed cake to kill climbing perch within 48 hours. The results
showed that limitation of the 48-hr LC50 concentration at 95% confidence of tea seed cakeon climbing perch (size of approximately 5±2 cm) was 33 (31.16-36.10) ppm. The slope
function was 1.361(0.88-2.11) ppm and safe concentration was 1.65 ppm. This result canbe used to apply for controlling of fish predator as fish which have accessory respiratoryorgans for farming application.
Keywords acute toxicity, tea seed cake, climbing perch

INTRODUCTION
Fish predator is one problem of aquaculture farming because it can lead to bring about disease, thedisplacement of native species, and causing changes quality of product. Similarly, it can cause of
decreasing of the production due to raise or mortality. Climbing perch (Anabas testudineus(Bloch)) often found in the shrimp pond or fish farming. It is an omnivorous species (Trung, 1999)
and consumes various kinds of food such as detritus, aquatic plants, crustaceans, worms, mollusksand insects. Thus, it has been described as carnivore or an insectivore and plays a role of predator(Yakupitiyage et al, 1998). In addition, it is a fast growing species which is able to live in deep orshallow water and can survive in low oxygen condition (Khatune-Jannat et al., 2012). It is also able
to migrate between ponds (Trieu and Long, n.d.). Therefore, large numbers of climbing perch inponds cause slow growth rate or decrease in average productivity. The farmers have tried toeradicate climbing perch with chemicals but many are harmful to the products and also users. Tea
seed cake is the residue of Camellia sp. seeds after oil extraction that contains 5.2-7.2% of saponin(Minsalan and Chiu, 1986) which is a toxin giving haemolysis of blood. Owing to the eradicableeffectiveness of the tea seed cake, many farmers apply it to eliminate predator fishes, shellfish or
tadpoles in the fish and shrimp ponds. The effective dosage of crude saponin for eradicating
predatory fishes depends on the proportion of body weight, pond’s salinity level (Terazaki et al.,Ⓒ ISERD126

IJERD – International Journal of Environmental and Rural Development (2014) 5-21980) and fish species. In order to provide a guidance of using the tea seed cake, the study of its
acute toxicity on climbing perch (A. testudineus (Bloch)) is useful for farming application.
OBJECTIVE
An objective of the study was to determine the median lethal concentration of tea seed cake thatwould result in the eradication of climbing perch (A. testudineus (Bloch)) at 48 hours.
METHODOLOGY
Animal preparation: One thousand climbing perch (A. testudineus (Bloch)) (size ofapproximately 5 ± 2 cm) were transported from climbing perch farm in central Thailand tolaboratory. They were reared in 500 liter fiber glass tanks that was contained treated fresh waterand added aeration for 7 days to acclimatize before bioassay testing. Food did not add over for 24hr
before the experiment and throughout testing period.
Chemical preparation: Power tea seed cake that contains saponin 12% as active ingredient is usedin the experiment. The tea seed cake solution was prepared in distilled water to dilute concentration
for range finding test and definitive test. Each tea seed cake concentration was added one time at
the beginning of each test.
Acute toxicity of tea seed cake to climbing perch: Static bioassay method was used to assess thetea seed cake concentration that caused 50% death of climbing perch within 48 hr test period (48-hrLC50) which consist range finding test and definitive test (Rand, 2003).
Range finding test was the determination of the range concentration from the lowest that 100%of test animals dead to the highest that 100% of test animals survived. Variation of the tea seedcake concentration was added at 8 liter of water into 10 liter glass tanks which stocked 10 random
selected test animals, and then each treatment was run three replications. Fish were observed andrecorded the number of mortalities for 48 hr when they did not have operculum movement and did
response while touching with a glass rod. The result of concentration in the first step was used todetermine the further concentration in the next step. The experiment for the definitive test wasdetermination of concentration range from the result of previous step on logarithmic scale. The
behavioral response and mortality of climbing perch were recorded within 48 hr. Dead fish wasimmediately removed after recorded.
Temperature, pH and dissolved oxygen were determined with YSI 550A and Sartorius Basic
pH Meter PB-11 before and after toxicity test.
Data analysis: 48-hr LC50 values and slope function were calculated at 95% confidence limits
according to Litchfield and Wilcoxon (1949). Data were plotted on logarithmic-probability paper.
RESULTS AND DISCUSSION
No mortalities were observed in controls and concentration of 19 ppm concentration of tea seedcake during the 48 hr test period. However, mortalities started within 18 hr in the concentration of
25.69 and 34.74 ppm. Meanwhile, the 47 ppm caused mortality of test fish approximately
10.00±1.00% after 6 hr exposure and 100% after 48 hr in all replications. The mortalities wereobserved increasingly in the high concentration during almost the end of experiment (Table 1). Inaddition, the 33 ppm (31.16-36.10) was calculated as the 48-hr LC50 with 95% of confidence limit
for climbing perch (A. (Bloch)) of size approximately 3-7 cm in the tea seed cake (Table 2). The
result showed high concentration when it was compared with general concentration for usage infresh or low saline water at 25-30 ppm (Hongkanarat et al., 2011). This is probably due to thetolerance levels of climbing perch in the hard conditions that is an ability to survive in a wide rangeof chemicals and temperatures owing to its accessory respiratory organs such as a labyrinth organ.
The slope function and safe concentration showed that 1.361 (0.88-2.11) and 1.65 ppm,
respectively.
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IJERD – International Journal of Environmental and Rural Development (2014) 5-2

Table 1 Accumulative mortality percentage of climbing perch (A. testudineus (Bloch))(mean±SD) after tea seed cake exposure
Duration ofexposure to tea
seed cake (hr.)
06
12
18
24
30
36
42
48

% Accumulative mortality
0 ppm

19 ppm

25.69 ppm

34.74 ppm

47 ppm

0.00±0.00
0.00±0.00
0.00±0.00
0.00±0.00
0.00±0.00
0.00±0.00
0.00±0.00
0.00±0.00
0.00±0.00

0.00±0.00
0.00±0.00
0.00±0.00
0.00±0.00
0.00±0.00
0.00±0.00
0.00±0.00
0.00±0.00
0.00±0.00

0.00±0.00
0.00±0.00
0.00±0.00
3.33±0.58
10.00±0.58
13.33±0.58
16.67±0.58
20.00±0.58
23.33±0.58

0.00±0.00
0.00±0.00
0.00±0.00
30.00±0.58
36.67±1.15
40.00±1.15
40.00±0.58
43.33±0.58
43.33±0.58

0.00±0.00
10.00±1.00
16.67±0.58
36.67±1.15
83.33±1.15
96.67±1.53
96.67±0.00
96.67±0.00
100.00±0.00

Table 2 48-hr LC50 values and slope function with lower and upper limit of 95% of tea seedcake on climbing perch (A. testudineus (Bloch))
48-hr LC50(ppm)

slope function

33 (31.16-36.10)

1.361(0.88-2.11)

In addition to the mortality, exposed climbing perch to the tea seed cake also showed increasein excretion and released more mucus in high concentration treatment. The nearly died fish has an
opaque body color compared with normal fish and slow response or movement. For those ofmortality experiencing, no movement of fin and operculum at all and they were observed at thebottom of the ponds. The response is in agree with El-Murr et al. (2014) that is likely due toreflection increase of oxidative stress that showed revealed normocytic normochromic anemia andsignificantly increases level of SOD and CAT activities and gene expression. The damaged ofgeneral tissues in various organs was also detected with high doses of tea seed cake (El-Murr et al.,2014). According to Boyd (1990), saponin compound in tea seed cake could destroy erythrocytes
and was highly toxic to fish. Also saponin acts chiefly with lowering the surface tension between
water and gills of fish and induces haemolysis, so preventing oxygen uptake by fish and leading toslow dead by oxygen deprivation (Lamba, 1970). Moreover, Roy and Munshi (1989) reported thatthe perch A. testudineus that were exposed for 24 hr with 2-5 mg/l saponin showed increasing of
20% oxygen uptake and increased its number of erythrocyte, amount of hemoglobin andhematocrit.
Temperature and pH in all treatments are not different before and after test. Meanwhile
dissolved oxygen showed optimum level before the test and decreasing later after the test (Table 3).
Nevertheless, these parameters are not likely a cause of climbing perch mortality because the fish
can thrive well in deficient dissolved oxygen waters. Moreover, it has better ability of breathing
due to its special organs and tolerance of adverse environmental conditions (Trieu and Long, n.d.).
For this reason, the fish mortality in the experiment is more likely caused by the tea seed caketoxicity.
Table 3 Temperature, pH and dissolved oxygen levels before and after toxicity test
Concentrationof tea seedcake (ppm)
0.00
19.00
25.69
34.74
47.00

Temperature (๐C)

pH

Dissolved oxygen (mg/l)

BeforeAfter

BeforeAfter

BeforeAfter

28.1±0.00
27.9±0.00
27.9±0.00
27.8±0.00
27.9±0.00

28.3±0.10
28.3±0.06
28.3±0.06
28.2±0.06
28.2±0.06

7.7±0.00
7.7±0.00
7.6±0.00
7.7±0.00
7.7±0.00

7.5±0.00
7.5±0.00
7.5±0.00
7.5±0.00
7.6±0.00

5.08±0.29
5.01±0.25
4.95±0.09
5.33±0.27
5.07±0.14

2.69±0.16
2.48±0.09
2.21±0.06
2.17±0.08
1.27±0.19

However, different fish species showed different tolerances. The species as highest mortality
in 24 hr of tea seed cake exposure was observed in Puntius gonionotus, followed by CyprinusⒸ ISERD128

IJERD – International Journal of Environmental and Rural Development (2014) 5-2

carpio, Gambusia sp. and Clarias sp., whereas tea seed cake toxicity ended after 12 hr(Chiayvareesajja et al., 1997). Terazaki et al. (1980) reported that the toxicity of saponin weakened
with time. In agree with, Hongkanarat et al. (2011) reported that 48-hr LC50 of tea seed cake on
Pacific white shrimp (Litopenaeus vannamei) post larvae 10 (PL10) was 416.14 ppm at salinity of30 ppt that showed less toxicity to shrimp larvae. The results of this experiment showed that tea
seed cake could be use to control climbing perch in farms. Further study is suggested to study moreon the toxicity to other aquatic species for farming application.
CONCLUSION
The study indicates that tea seed cake is a natural product that can be used for eradication ofclimbing perch (A. testudineus (Bloch)) in aquaculture farming. The solution is very applicable
control of fish having accessory respiratory organs. Moreover, it is environmental friendly usedwhen apply it following the guideline and thus, giving sustainable utilization.
AKNOWLEDGEMENTS
The authors would like to thank Faculty of Agricultural Technology, Rajamangala University ofTechnology Thanyaburi which was supported some budget in this experiment.
REFERENCES
Boyd, C.L. 1990. Water quality in ponds for aquaculture. Birmingham Publishing Co., Birmingham, AL,
482.
Chiayvareesajja, S., Rittibhonbhun, N., Hongpromyart, M. and P. Wiriyachitra. 1997. Toxicity of the Thai
piscicidal plant, Maesa ramentacea, to freshwater fishes in ponds. Aquaculture, 158, 229-234.
El-Murr, A., Ali, H.A. and Eldeen, N.A.M.N. 2014. Molecular, biochemical and histological effects of tea
seed cake on different organs of Oreochromis niloticus. Global Veterinaria, 13 (5), 711-719.
Rand, G.M. 2003. Fundamentals of aquatic toxicology: Effects environmental fate and risk assessment.
Second Edition. Taylor & Francis Ltd., USA., 1120.
Khatune-Jannat, M., Rahman, M.M., Bashar, A., Hasan, N., Ahamed, F. and Hossain, Y. 2012. Effects ofstocking density on survival, growth and production of Thai climbing perch (Anabas testudineus) underFed Ponds. Sains Malaysiana, 41 (10), 1205-1210.
Lamba, S.S. 1970. Indian piscicidal plants. Econ. Bot., 24, 134-136.
Hongkanarat, L., Limsuwan C. and Chuchird, N. 2011. Acute toxicity of tea seed cake on Nile Tilapia(Oreochromis niloticus), giant seaperch (Lates calcarifer) and Pacific white shrimp (Litopenaeusvannamei), Proceedings of 49th Kasetsart University Annual Conference: Fisheries, Bangkok, Thailand,
49-56.
Litchfield, J.T. and Wilcoxon, F. 1949. A simplified method of evaluating dose-effect experiments. J.
Pharmacol. Exp. Ther., 96 (2), 99-113.
Minsalan, C.O. and Chiu, Y.N. 1986. Effects of tea seed cake on selective elimination of finfish in shrimp
ponds. In; J.L. Maclean, L.B. Dizon and L.V. Hosillos (Editors), The First Asian Fisheries Forum. AsianFisheries Society, Manila, Philippines, 79-82.
Roy, P.K. and Munshi, J.P. 1989. Effect of saponin extracts on oxygen uptake and hematology ofanbreathing climbing perch, Anabas testudineus (Bloch). J. Freshwater Biol., 1, 167-172.
Terazaki, M., Thambuppa, P. and Nakayama, Y. 1980. Eradication of predatory fishes in shrimp farms inutilization of Thai tea seed. Aquaculture, 19, 235-242.
Trieu, N.V. and Long, D.N. 2001. Seed production technology of climbing perch (Anabas testudineus): A
study on the larval rearing. Institute for Aquaculture and Fisheries Sciences, College of Agriculture, Can
Tho University.
Trung, N.M. 1999. Some biological characteristics and reproduction of climbing perch (Anabas testudineus
Block). Master thesis in Nha Trang Fisheries University. Vietnam.
Yakupitiyage, A., Bundit, J., and Guttman, H. 1998. Culture of climbing perch (Anabas testudineus): Areview. AIT Aqua outreach. Working Paper, New Series No. T-8, 9.

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