ISSN - 1811-8976

JOURNAL OF

SUBTROPICAL

AGRICULTURAL RESEARCH

AND DEVELOPMENT

J. Subtrop. Agric. Res Dev. 2(2): 83-89, September 2004

PESTICIDAL PROPERTIES OF SOME INDIGENOUS PLANT EXTRACTS ON Penaeus

monodon (PENAEIDAE)

M. S. RAHMAN', A.S.M. SAIFULLAH^ S.M.A. JABBER^ and Y.S.A. KHAN'

ABSTRACT Alcoholic extracts of three plants, viz., Sapium indicum, Euphorbiaceae (Willd.), Abrus precatorius, Leguminosae (Linn.) and Baningionia acutangula, Lecythidaceae (Gaertn.) were tested on Penaeus monodon (Penaeidae) under laboratory conditions. P. monodon was found highly susceptible when exposed to the extract of the fruits of S.indicum (Willd.). No toxic response was observed for the other two extracts when the shrimp species was exposed to a test dose level of 3000 mg 1 ' for 48 hours. The LD50 value of the water-soluble absolute alcoholic extract of the fruits of S.indicum after 12 hrs P. monodon was found 788.86mg 1 '. The x2 value was found significant at 1% level of significance. The present study reveals that the water-soluble absolute alcoholic extract of the fruits of S. indicum may be useful as pesticide, especially in the shrimp culture ponds, remaining the targeted species unharmed.

Key words: Sapium indicum (Willd.), Abrus precatorius (Linn.), Barringtonia acutangula (Gaertn.), Penaeus monodon, Piscicide, Toxicity

INTRODUCTION

Bangladesh has a vast and diversified fishery potential in its fresh, brackish and marine waters. The country has more inland fish species than all of Europe (Minkin et. al, 1997). Pollution resulting from the extensive use of pesticides and fertilizers not only threatens the survival of fish resources but also greatly reduces the amenity of the fishing communities ( lUCN, 1993). However, alternative means to reduce the use of chemicals in aquaculture have been proposed by several authors (Lightner et al, 1992, Keachum, 1994 and Lin, 1995). The amount of information on chemical use in coastal aquaculture and its significance to human health, environmental protection, and sustainable development of the sector has increased over the last decade (GESAMP, 1997).

About 22 species of fish have so far been identified to prey on shrimp, Penaeus monodon (Siddiquee, 1984 and Naser et al, 1992) and various methods are used to control undesirable fish in shrimp ponds (GESAMP, 1997). Though a number of plants have been tested scientifically on some freshwater fish species (Latifa et al, 1992, 1997; Latifa and Begum, 1993 and Nasiruddin et al, 1997) for exploring piscicidal effectsof plant extracts. But tests on marine species are quite rare except Sin (1989).

Sapium indicum (Willd.), Abrus precatorius (Linn.) and Barringtonia acutangula (Gaertn.) are common in Bangladesh (Yusuf et al, 1994) and are well known for their insecticidal properties. The present work was therefore undertaken to assess the effects of the fiiiits of S. indicum (Willd.), seeds of A. precatorius (Linn.) and bark of 6. acutangula (Gaerm.) under laboratory conditions.

MATERIALS AND METHODS

Extraction of test substances from the fruits of 5. indicum (Willd.) Fresh fruits (10kg) were collected from Nayarhat village of Chittagong district. The fruits were

chopped into small pieces and dried. The dried substance was pulverized into fine granules with the help of a power-driven grinder. The granular matter was extracted with lOL of petroleum ether (40" -(><fC) for three days. The crude extract was filtered and evaporated through a rotary vacuum evaporator at 50 + 5° C. The crude extract thus obtained was collected in a conical flask (250 ml). Solvent i f any

'institute of Marine Sciences, University of Chittagong, Chittagong-4331, Bangladesh, 'Mawlana Bhashani Science and Technology University, Santosh, Tangail-1902, Bangladesh.

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M. S. Rahman, A S M . Saifullah, S.M.A. Jabber and Y.S.A. Khan

remaining in the crude extract was removed with the help of an exhaust pump. The solvent free exfract was then stored in a refrigerator.

The residue remained on the filter paper was dried and extracted again with alcohol (9L) for three days. Then the solvent free crude extract was obtained by following the same procedure as illustrated earlier. The crude extract was then subjected to the serial fractionation as illustrated by Sharma and Simlot (1971). The final fi"actions (water soluble and alcohol soluble extracts) were collected into two separate flasks (250 ml and 50 ml). The extracts were dried in an oven for six hours at temperature 30 ± 5" C and stored in a refrigerator. ^

Extraction of test substances from the seeds of ^. precatorius (Linn.) Freshly dried seeds (5kg) of the plant were collected from a local market (Terry Bazar,

Chittagong). The same methodologies were applied for obtaining petroleum ether (40''-60''C) extract as well as water-soluble and alcohol soluble extracts. A l l the extracts were stored in a refrigerator.

Extraction of test substances from the bark of B. acutangula (Gaertn.) Fresh bark (10kg) of the plant was collected from the creek of Wazedia village of Chittagong

district. The same procedure was followed for obtaining different extracts as before. The extracts were stored in a refrigerator.

Collection of shrimp and seawater Shrimp species, Penaeus monodon was collected from coastal fishermen of Cox's Bazar and

kept in a fiberglass container (15L) containing seawater (P" 7.2 and salinity 33 ppt.). They were gradually acclimatized in seawater (P" 6.8 and salinity 20 ppt.) for 24 hours. They were then observed for two days and then only the healthy and normal ones were subjected to the subsequent experiment. Seawater was collected from Cox's Bazar seacoast, the Bay of Bengal. The seawater was kept in a fiber wood tank (3 tons) and was allowed to settle the particulate or other suspended matters for 48 hours. Then the seawater was transferred to another tank (3 tons) with a siphon. Aeration was done to increase dissolve oxygen (DO) of the stocked water.

Measurement of physical and chemical data Salinity, temperature, DO and P" of the seawater were measured with the help of salinity

meter, thermometer, Winkler's method (APHA, 1981) and digital P" meter respectively. Total length of the shrimp was measured by using a millimeter scale.

Toxicity test on Penaeus monodon In the present experiment, only the water-soluble alcoholic extracts of the three plant samples

were used to evaluate their toxic effects on P. monodon under laboratory conditions. Toxicity test of the extracts was conducted by following the Static Bioassay Method (APHA, 1981). For each of the extracts, five concentrations were prepared for the explorative test. Definitive test was then carried out for the effective test substance(s). In the definitive test, five specimens of P. monodon (total length 29 ± 2 mm) were immersed in each glass jar (lOL, dia 13.5cm) for each concentration of the effective test substance with the seawater (P" 6.8 and salinity 20 ppt). Five concenttations were made for each of the effective test substance while two replicate glass jars (10 specimens) was used for each of them. A control group was also maintained. Normal food was given to the shrimp during the experimental time. Water parameters (temperature, DO, P" and salinity) were measured before and after each experiment.

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Pesticidal properties of some indigenous plant extracts on Penaeiis monodon (penaeidae)

Statistical analyses A l l the data derived from the experiment were analyzed statistically. Statistical analyses were

done on the Probit Analysis, regression line equation (Finney, 1971), -test and F-test (Gupta and Kapoor, 1994). The calculated values o f the tests were compared with The Tables o f the Statistics for

(n-1) degrees o f freedom at 1% level o f significance (Fisher and Yates, 1963).

RESULTS AND DISCUSSION

From the present study it was found that the water-soluble alcoholic extract o f the fruits o f Sapium indicum (Wil ld . ) was the effective extract in k i l l ing s h r i n ^ while the other two extracts caused no mortality o f shrimp even at the highest test dose level (3000 mg 1 ') for 48 hours exposure. F rom the probit mortality analysis it was found that the 12-hr L D 5 0 value o f the water-soluble alcoholic extract o f the fruits o f S. indicum (Wil ld . ) for P. monodon was 788.86 mg 1 ' with the fiducial limits o f 788.86 ± 1.28 (mg 1 ') at 1% significance level. Regression line for the probit mortality data has been shown in the Figure 1. The regression equation for the probit mortality data was found as Y = - 9.823 + 5.117X where X and Y stand for the log concentration o f the test substance and the probit mortality o f P. monodon respectively. The value for the percentage mortality o f shrimp was found highly significant at 1% level. The two-way analysis o f variance at 1% level o f significance showed that variation between treatments was insignificant while variation between replications was significant.

Information on toxicity o f pesticides, in general, is abundant but not much seems to be known for the mortality of shrimp, P. monodon. B y the present time some works have been carried out with different form of extracts o f S. indicum and B. acutangula to find out the ichthyotoxicity, which are mainly concerned with freshwater hardy fish species. The studies showed that the 24-hr LD50 value o f the seed karael o f S. indicum for Heteropneustes fossilis ranged from 381.646 ppm to 805.061 ppm (Chowdhury, 1996) and 18.62ppm for the dry fruits (Kha l i l , 1984); the 8-hr LD50 value o f the fresh root bark of 5. acutangula (Gaertn.) for Heteropneustes fossilis (Bloch) was found 245.47 ppm whereas it was 186.20 ppm for the dry root bark (Akter, 1994).

From the present study it was revealed that P. monodon is capable o f tolerating a large range o f toxicity than H. fossilis when exposed to the water-soluble alcoholic extract o f the fiiaits o f 5. indicum (Wil ld . ) . For eradicating weed and predatory fishes from the culture ponds, extractive product o f S. indicum (Wil ld.) may be used safely remaining the targeted species (P. monodon) unharmed as there is an evidence that some o f the common predatory fishes in the coastal shrimp ghers/ponds are less tolerable to the extractive products o f Denis eliptica and Thea sinensis i n the range o f 0.05 to 10.00 ppm (Sin, 1989).

The ftxiits o f S. indicum contain saponin (Chopra et al., 1992). The lethal concentration o f saponin for fish and crustaceans remains in a ratio o f as widely as 1:50 (Tang, 1961). F rom this calculation it is estimated that the lethal concentration o f the water soluble alcoholic extract o f the fruits o f S. indicum (Wil ld.) for fish should be 15.8 m g . l ' which is similar to the work done by K h a l i l (1984). The fruits o f the S. indicum was found to be more toxic than the spruce bark extract as the 96-hr LD50 values of the later were recorded as 415 and 205 m g . l ' for the larval and adult pink shrimp, Pandalus borealis respectively (Buchanon et al., 1976).

P. monodon when exposed to different concentrations o f water-soluble alcoholic extract o f the fruits o f S. indicum, normal behaviour was observed for the first one hour. Abnormal behaviour was observed within two hours. The shrimp at times were found to come up to the surface water following the direction o f water movement resulting from continuous aeration. This phenomenon was observed to continue for the first four hours o f observation. Wi th in six hours, the shrimp was found drawing upwards, but to the half o f the previous distance. Wi th in eight hours, body colour was seen to change into whitish and the test species were found to loose their locomotory activity. After ten hours, they

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M. S. Rahman, A.S.M. Saifullah, S.M.A Jabber and Y.S.A. Khan

were found to be non-motile, lying at the base of glass jar where water movement was comparatively lower than the middle or upper layer o f water column. Wi th in 12 hours, deshelling was observed in some species along with a pale body colour that was associated with the higher concentrations (>1000 mg.r') o f the test substance.

7 T 1

Fig 1: Regression line for determining the 12-hr LD50 value o f the water-soluble absolute alcoholic extract o f the fruits o f Sapium indicum (Wil ld . ) for penaeus monodon

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Pesticidal properties of some indigenous plant extracts on Penaeus monodon (penaeidae) - :

The toxicant o f the fnaits extract o f S. indicum (Wil ld . ) seemed to effect the neuromuscular coordination, as the shrimp was found to loss their locomotory power and settled on the bottom prior to death. The shrimp became immobil ized within six to eight hours o f observation. Similar neurotoxic effect o f the plant, Terminalia bark sap was observed in the h i l l stream fish, Garra lamta by Ojha et al. (1989).

The principal poisonous constituents o f the seed of Abrus precatorius (Liiui . ) are abrin (Dymock et al, 1892) and abrulin (Khan et al, 1961). In the present study, the percentage occurance o f the toxic principles in the seeds of A. precatorius (Linn.) was probably low as there was no remarkable change in the test species of shrimp. The underlying reason(s) might be o f the quality o f the seeds used and/or environmental conditions as these are known to affect the chemical characteristics o f the seeds (Khanefa / . , 1970a).

The seed powder o f B. acutangula at a concentration o f 20 ppm was also reported to k i l l a wide variety of fish (Chakraborty et al. 1972). But in the present study, bark o f B. acutangula (Gaertn.), which was reported to contain 16% tannin (CSIR, 1948), was foimd to k i l l no shrimp species at the highest test dose of 3000 mg 1"'.

Through several investigations it is comprehended that toxic chemicals o f plant origin are the best type o f poison for k i l l ing fish from the culture ponds in regards to synthetic and long persisting pesticides as the toxic chemicals o f plant origin are selective as well as they have shorter retention time in the environment due to their rapid biodegradability. For instances, toxic effect o f dry root bark extract and seed powder of B. acutangula lasts in the envirorunent for five days (Akter, 1994) and two days (Chakraborty et al.. 1972) respectively after application. In this regard, fruits o f 5. indicum (Wil ld . ) may by far, an ideal piscicide o f plant origin.

In the present study water temperature before and after the application o f the extracts were 29±l ' 'C and 28± l ' 'C respectively, which was mainly due to the cooling effect resulting from continuous aeration. A rise in temperature was observed by others (Parvin, 1986; Ahsan, 1989; Akter, 1994 and Khal i l , 1984), as they followed no aeration. They also found a decrease in P " value o f water during the experimental hours. The same phenomenon was observed during the present experiment where P " value was found to decrease within the range of 0.1 to 0.3 after each experiment. This might be due to the excretion of metabolic wastes by the shrimp species as well as decomposition o f the supplied food material. Salinity was found 19.8 ppt and 20 ppt before and after the present investigation that was probably due to the evaporation of water. Dissolve oxygen was found to increase in the range of 0.19 to 0.23 mg l ' from the initial concentration of 8.59 mg P ' which might be due to continuous aeration o f water during the experimental hours.

Natural plant resource is a viable source of organic pesticides as they are friendly to environment for not posing and threat to any of its components. To be too friendly to our environment, dependence on and use of environmentally harmful agents especially synthetic pesticides, may be reduced by using namral plant-based products through exploring them in a sustainable manner. In this regard, fruits o f S. indicum (Wil ld . ) may be used to reduce or combat the use o f synthetic pesticides for eradicating predatory fishes from culture or nursery ponds.

ACKNOWLEDGMENT

The authors express their gratitude to M r . M d . A b u l Khair , Deputy Director and M r . G o l a m Mustafa, P.S.O., Fishery Research Institute, Cox ' s Bazar, Bangladesh for their assistance during the present work.

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