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Acute Toxicity of Organophosphate and Synthetic Pyrethroid Pesticides to Juveniles of the Penaeid Shrimps, Metapenaeus monoceros.

Byline: Nafisa Shoaib and Pirzada Jamal Ahmed Siddiqui


The present study examined the acute toxicity of organophosphate pesticides (methyl parathion, chlorpyrifos) and synthetic pyrethroid pesticides (fenvalerate, fenpropathrin) to shrimp juveniles (Metapenaeus monoceros Fabricius, 1798). The 24 h LC50 for penaeid shrimp juvenile was 0.1, 1.3, 0.65 and 0.26 ppb for chlorpyrifos, methyl parathion, fenvalerate and fenpropathrin, respectively. This high sensitivity of juveniles to pesticides is alarming as it may have implications on the fishery industries which play a significant role in the national economy and towards the food security of the country.

Key words: Pesticides, methyl parathion, fenvalerate, shrimp juveniles, ecosystem.


The indiscriminate use of agrochemicals to protect crops from insect pest has increased from past decades. Pesticides could contaminate land and water from production sites and storage tanks, run-offs from fields. Pesticides are washed into surface waters and because of its aquatic distribution, it affects a wide range of non-target organism like invertebrates, mammals, birds and fishes, especially those inhabiting the marine environment (Burkepile et al., 2000; Selvakumar et al., 2005). Organophosphate (OP), carbamate and synthetic pyrethroid (SP) pesticides are mostly used in agriculture to control pests (Kumar et al., 2010) due to their non-persistent nature in the environment. Although these pesticides rapidly degrade their high acute toxicity to some non-target species has been demonstrated in many laboratory tests (Abdullah et al., 1994; Olima et al., 1997; Phyu et al., 2004, 2005; Sial et al., 2009).

Laboratory studies have shown that pesticides can be acutely toxic to estuarine organisms including crustaceans (Goodman et al., 1988; Randall et al., 1979; Ringwood, 1993; Bhavan and Geraldine, 1997, 2001; Suryavanshi et al., 2009; Shoaib et al., 2012a). The penaeid shrimp, Metapenaeus monoceros (Fabricius) is one of the economically and nutritionally important shrimp species that inhabits the mangrove swamps in coastal areas of Pakistan. Metapenaeus monoceros are locally named as 'jaira' attaining size of 190 mm. Shrimps are active animals so the symptoms of insecticidal stress are easily detectable. In line with other decapod crustaceans, shrimps also have drastic effect of pesticides (Babu et al., 1987; Sanders and Cope 1966; Bhavan and Geraldine, 1997, 2001; Suryavanshi et al., 2009; Tu et al., 2012).

Acute toxicity bioassays are a convenient tool used extensively to assess the toxicity of physiologically active substances and also to evaluate the potential of chemical contamination on commercially and ecologically important species (Ahsanullah and Arnott, 1978). Penaeid shrimps (Metapenaeus monoceros Fabricius, 1798) were selected for bioassay experiments. The objectives of the present study was to assess and compare the acute toxicity of OP and SP pesticides to shrimp acting individually.


Preparation of chemicals

Pesticides, methyl parathion 5% EC, fenpropathrin 20% EC, fenvalerate 20% EC, chlorpyrifos 40% EC were procured from Pakistan Agricultural Research Center. Stock solution of 100 ppm and appropriate working concentrations were prepared in filtered seawater.


The shrimps (Metapenaeus monoceros) juveniles were collected from Sandspit backwaters (mangrove area) using handnet. The sandspit backwater provide habitat for wide variety of vertebrate and invertebrate species, and is considered an important spawning ground. The adjacent areas comprise of a mangrove forest which provides a highly productive environment conducive for sustaining the diversity in the area. From the sandspit the shrimps were transported in clean aerated seawater to the laboratory ensuring minimum stress and acclimatized in the laboratory for one to two weeks (Ahsanullah, 1976; Krishnakumar et al., 1987) in glass aquaria (90cm length x 30cm width x 32 cm width) containing clean and aerated seawater at room temperature (281C), with salinity 30 ppt, pH 7.57, photoperiod 16 h light and 8 h dark were maintained throughout the acclimatization period. Shrimps measuring 2.381cm in length, and 1111 mg in weight were use in this study.

The shrimps were fed ad libitum to avoid cannibalism. Seawater in the aquaria was replenished everyday in order to maintain the water quality. The organisms were not fed at all during the experiments.


Standard bioassay methods (APHA, 1971) were followed to evaluate toxicity of pesticide using static bioassay system (Doudoroff et al., 1951). Bioassays to evaluate LC50 were carried out in glass jars (20.5cm length x 13.5cm width) of 2 liters capacity for shrimp juveniles. All glassware was acid cleaned prior to the tests. Initially all test organisms were treated with wide range of pesticide concentration in filtered seawater to evaluate the concentration at which mortality around 50% occurs. For each concentration of test pesticide ten shrimp juveniles were exposed in groups. The experiment was repeated with five or more concentrations of different pesticides for test organism. The different concentrations of pesticides ranged between 0.1-8 ppb. The tests and controls for each experiment were in triplicate and the controls had only seawater.

The other experimental conditions, such as, temperature (281C), Salinity 30 ppt, pH 7.57, photoperiod 16 h light and 8 h dark were maintained throughout the experiment. Acute toxicity measured as mortality of organisms exposed to each pesticides was estimated by determination of the 24 h LC50 (the concentration of the pesticides which kills 50% of the test animals after 24 h exposure). Organisms were considered dead if they did not exhibit any internal or external movement and it laid immobile. The LC50 values were determined by using computer programme, Biostat 2009 based on Finney Method 1952 (Probit analysis).


The results obtained in experiments, where shrimps were tested against organophosphates and synthetic pyrethroid pesticides, show that shrimp were sensitive to all pesticides tested (Table I). The rate of mortality (%) was directly proportional to the concentration of pesticides. The variability in the degree of sensitivity is reflected by the lethal concentration values of pesticides, at which 50% mortality occurs (Fig. 1). The 24 h LC50 was 0.1, 1.3, 0.65 and 0.26 ppb for chlorpyrifos, methyl parathion, fenvalerate and fenpropathrin respectively. Among OP, shrimp juvenile are considered as more sensitive to chlorpyrifos than methyl parathion where as among SP, shrimp juvenile are more sensitive to fenpropathrin than fenvalerate.

Shrimp appear to be highly sensitive to pesticides and have low LC50 values, which may be due to the fact that both OP and SPs are particularly produced to target insects (Ankley and Collyard, 1995; Flemer et al., 1997; Pesando, 2003) which also include crustaceans. As OP and SP pesticides are non-persistent in nature and readily degradable, therefore acute toxicity test of 24h LC50 were considered in the present study.

Juveniles used in the present study, were highly sensitive as expected in line with findings of some previous studies, in other organisms (Hart et al., 1991; Hall and Burns, 2002; Boateng et al., 2006) for example, copepods, (Andrew et al.,1995), brine shrimp (Sanchez-Fortun et al., 1995), fish (Bansal et al., 1980; Shoaib et al., 2012b, 2013), and mysids (Conklin and Rao, 1978; Goodman et al., 1988). Generally in bioassay juveniles are employed for toxicity test to predict environmental risk. Juveniles are more sensitive to environmental impacts than the adult (Warren et al., 1995; Fisher et al., 1999; Kefford et al., 2004). The reason is that juveniles have higher surface area to volume ratios than adults and have faster uptake kinetics of the chemical (Kefford et al., 2004). The metabolic capacity in juvenile shrimp may be different from that in adults (Sucahyo et al., 2008).

Table I.-Toxicity of organophosphate pesticides and synthetic pyrethroid pesticides on Metapenaeus monoceros after 24 h of treatment showing LC50.

###Pesticides###No. of shrimps###Concentration tested (ppb)###LC50 (ppb)###Intercept###- square###p-level

Methyl parathion###150###1.0-8###1.30.06###14###0.94###0.82




The LC50 values found in the present work can be compared with those reported earlier for peneaid shrimp. LC50 values obtained by other authors for effects of different pesticides on various shrimp species are presented in Table II. The comparison of LC50 values for different organisms provides only a rough indication of differences in specific tolerance as a number of factors influence the bioassay results, such as, temperature (Macek et al., 1969) and degree of susceptibility of test organisms (Macek and McAllister, 1970). A wide range of pesticides have been found to increase the toxicity at higher temperature (Macek et al., 1969; Muirhead-Thomson, 1971). For any species, sensitivity to a given pesticide varies with age, sex, nutritional background, health, stress and the environment (Sanchez-Fortun et al., 1995).

Variability due to differences in sensitivity between sexual and asexual species, as well as among intra-strains and clones of the same species has been reported for aquatic invertebrates used in ecotoxicological studies (Baird et al., 1990; Moller et al., 1996). It is however evident that the toxicity differs from species to species (Pickering et al., 1962; Boateng et al., 2006; Shoaib et al., 2012a) and in some cases from place to place, which may be due to differences in bioassay techniques and purity of pesticides used, the differences found could also result from differences in tolerance to the exposure between species or populations of the same species (Chambers and Yarbrough, 1974). Different pesticides or even different salts of same pesticide have variable effect on same organisms (Tooby et al., 1975; Babu et al., 1987; Shoaib et al., 2012a).

This is also true for the toxicity of same group of pesticide to the same organism e.g. peneaid shrimp when exposed to OP pesticide; phorate and methyl parathion (Butler, 1964), when exposed to organochlorine (OC) pesticide; DDT and heptachlor (Chin, 1961; Butler, 1963), when exposed to OC pesticide; BHC and lindane (Schimmel et al., 1977). Together with previous studies, the present results support the use of M. monoceros as a sensitive bioindicator of pesticide contamination in the coastal environment.

Table II.-Effects of pesticides on various shrimp species in response to exposure showing lethal concentration (LC50) at which 50% mortality occurs.

Pesticide###Shrimp###LC50 (ppb)###Exposure time###Reference


Endosulfan###Litopenaeus stylirostris###230###48 h###Reyes et al. (2002)

DDT###Litopenaeus stylirostris###10790###48 h###Reyes et al. (2002)

DDT###Penaeus vannamei###8.7###48 h###Jose et al. (1996)

Chlordane###Penaeus vannamei###63.2###48 h###Jose et al. (1996)

Lorsban###Penaeus vannamei###4.8###48 h###Jose et al. (1996)

Lindane###Penaeus vannamei###3.9###48 h###Jose et al. (1996)


Methyl parathion###Litopenaeus stylirostris###38###48 h###Reyes et al. (2002)

Chlorpyrifos###Litopenaeus stylirostris###2260###48 h###Reyes et al. (2002)

Malathion###Litopenaeus stylirostris###34190###48 h###Reyes et al. (2002)

Fenitrothion###Penaeus japonicus###1###24 h###Kobayashi et al. (1990)

Fenitrothion###Penaeus japonicus###1.9###24 h###Lignot et al. (1997)


Permethrin###Palaemonetes pugio###0.10###24 h###DeLorenzo et al. (2006)

Permethrin###Litopenaeus stylirostris###290###48 h###Reyes et al. (2002)

The 24 h LC50 of shrimp juvenile was very low for both OP (chlorpyrifos, methyl parathion) and SP (fenvalerate and fenpropathrin) pesticides showing sensitivity of these organisms. Since Pakistan is an agricultural country and these pesticides are used in agricultural land. There is a paucity of data on the presence of pesticide in the coastal areas of Pakistan. Qasim et al. (1993) reported presence of low levels (pg g-1) of some pesticides in the bottom sediments of Korangi and Kodairo Creeks. Bano and Siddiqui (1991) reported slightly higher values of some of the OC compounds in sediment samples, from three locations along the Karachi coast (Korangi Creek, Manora channel and Hawks Bay) during low tide. Pesticides have lethal effect on shrimp and exposure to these chemicals may have adverse effects on growth, reproductive failure and/or death.

According to PEPA, the National Environmental Quality Standards (NEQS) relating to municipal and liquid industrial effluents for pesticide is 150 ppb (The Gazette of Pakistan, 1993). However, in our result the value of LC50 when exposed to pesticides is recorded as 0.1 to 1.3 ppb for shrimp, which is quite low. Serious ecological degradation may arise due to their potential to cause adverse effects on human and wildlife populations. The shrimp fishery industry is an important section of our national economy because of foreign exchange earned and employment produced from it. Deterioration in the quality of aquatic environment affects the shrimp fish industry as well as the aquaculture in coastal waters.


The authors would like to thank Dr Shahida Akhtar, ex. Director Pakistan Agriculture Research Centre for her guidance and valuable advice. The financial assistance provided by the Centre of Excellence in Marine Biology, University of Karachi is well appreciated.


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