Chinese Journal of Oceanology and Limnology   2017, Vol. 35 issue(1): 109-114     PDF       
http://dx.doi.org/10.1007/s00343-016-5173-3
Institute of Oceanology, Chinese Academy of Sciences
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Article Information

Md. SHAHJAHAN, Md. Farajul KABIR, Kizar Ahmed SUMON, Lipi Rani BHOWMIK, Harunur RASHID
Toxicity of organophosphorus pesticide sumithion on larval stages of stinging catfish Heteropneustes fossilis
Chinese Journal of Oceanology and Limnology, 35(1): 109-114
http://dx.doi.org/10.1007/s00343-016-5173-3

Article History

Received Jun. 5, 2015
accepted for publication Nov. 10, 2015
accepted in principle Dec. 20, 2015
Toxicity of organophosphorus pesticide sumithion on larval stages of stinging catfish Heteropneustes fossilis
Md. SHAHJAHAN1,2, Md. Farajul KABIR1, Kizar Ahmed SUMON1,3, Lipi Rani BHOWMIK1, Harunur RASHID1,4        
1 Department of Fisheries Management, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh;
2 Sado Marine Biological Station, Faculty of Science, Niigata University, Sado, Niigata 952-2135, Japan;
3 Department of Aquatic Ecology and Water Quality Management, Wageningen University, Wageningen University and Research Centre, P. O. Box 47, 6700, The Netherlands;
4 Science and Math Program, Asian University for Women, Chittagong 4000, Bangladesh
ABSTRACT: Sumithion is widely used to control brittle in paddy fields and tiger bug in fish larval rearing ponds. The objective of this study was to elucidate the toxic effects of sumithion on larval stages of stinging catfish Heteropneustes fossilis. Larvae were exposed to two concentrations (150 and 250 μg/L) of sumithion with one control in three replicates of each. Larvae samples were collected at 20- and 24-h intervals followed by observation under a digital microscope. Exposures of stinging catfish larvae to sumithion produced deformities including irregular head shape, lordosis, yolk sac edema, body arcuation, tissue ulceration, etc. The mortality rates of larvae were significantly increased in response to increase in sumithion concentrations. Furthermore, around 30% of the total adult stinging catfish reared in sumithiontreated aquaculture ponds were found to be deformed permanently. These findings highlight that exposure of stinging catfish to sumithion at the critical and sensitive stages in their life cycle may significantly reduce the number of returning adults. Therefore, the use of sumithion for crop protection needs to be considered carefully and alternatives to sumithion should to be developed for controlling aquatic insects in aqua-ponds during larval rearing.
Key words: environment     organophosphorus pesticide     fish     toxicity    
1 INTRODUCTION

In Bangladesh, numerous types of pesticides and herbicides are used for crop protection in agriculture. Over 98% of sprayed insecticides and 95% of herbicides reach non-target species, air, water, bottom sediments and food (Miller, 2004). Contamination of water by pesticides, either directly or indirectly can lead to fish kills that reduce fish productivity. Pesticides can affect fishes in a direct way through alterations of normal behavioral patterns, of physiology (e.g. sensorial, hormonal, neurological and metabolic systems), and of normal reproductive behavior (Clotfelter et al., 2004 ; Scott and Sloman, 2004). There are also some indirect toxic effects of pesticides on fishes through decreases in the fishes' food sources (algae and plankton), through changing food habits and through deterioration in the quality of aquatic habitats (Rahman et al., 2012 ; Cochard et al., 2014).

Sumithion, O, O Dimethyl O-(3-methyl-4- nitrophenyl), is an organophosphate insecticide that is widely used in aquaculture ponds to eradicate aquatic insects (mainly tiger bug) prior to release of larvae. It is also effective against a wide range of pests, i. e. penetrating, chewing and sucking insect pests, on cereals, cotton, orchard fruits, rice, vegetables, and forests. It may also be used as a spray for fly, mosquito, and cockroach protection for farms and public health programs. Sumithion is considered somewhat toxic to fish (Thomson, 1989). The 96-h LC50 was 1.7 mg/L for brook trout and 3.8 mg/L for bluegill sunfish (Meister, 1994), while the 48-h LC50 value for carp ranged between 2.0-4.1 mg/L (Worthing, 1987).

The toxic effects of pesticides can be observed by developmental, physiological and biochemical alterations in organisms (Salam et al., 2015 ; Sharmin et al., 2015). Embryonic and larval fishes are the most sensitive stages in their life cycles and are very sensitive to environmental pollutants (Marimuthu et al., 2013). These biomarkers can measure the interaction between environmental xenobiotic and biological effects. Inhibition and induction of these biomarkers are a good approach to measure potential impacts of pollutants on aquatic organisms (Miren et al., 2000).

Stinging catfish are native to Bangladesh, India, Pakistan, Nepal, Sri Lanka, Myanmar, Thailand, the Indus Basin and Laos including the Andaman Islands (Talwar and Jhingran, 1992) and are commercially important in Bangladesh. Although their main habitats are ponds, ditches, beels, swamps and marshes, they also occur in muddy rivers and slightly brackish water. Stinging catfish are capable of breeding in ponds and ditches during the onset of the monsoon when sufficient rainwater accumulates. Most of the catfish farms in Bangladesh are located in and around agriculture fields, or else their source of water is continuously in contact with the paddy ecosystem in which a lot of pesticides, herbicides and insecticides are used routinely. To the best of our knowledge, no earlier studies have been conducted to understand the toxicity of sumithion in fishes commonly found in the paddy ecosystem. In view of the aspects mentioned above, this study was aimed at assessing the toxic effects of sumithion on larvae of stinging catfish. The outcomes reported in this study will be useful for the management of the paddy ecosystem and improving the guidelines for sumithion use in rice fields and larval rearing ponds.

2 MATERIAL AND METHOD 2.1 Animals and chemicals

Healthy broods of stinging catfish were collected from a fish farm. They were reared and acclimatized in the ponds of the Faculty of Fisheries, Bangladesh Agricultural University until breeding was induced. The average body weights of male and female broods were 18.0 g and 38.8 g, respectively. Sumithion (60E/C) was collected from a local retail pesticide shop. The experimental procedures followed the guidance approved by the Animal Care and Use Committee of Bangladesh Agricultural University, Mymensingh, Bangladesh.

2.2 Induced breeding and fertilization of eggs

Five male and five female fishes were selected for breeding. Matured males were identified by a slightly pointed genital papilla, and females by a swollen abdomen and a reddish swollen vent. The maturity of each female was confirmed by slightly pressing the ventral side of the fish for oozing of eggs. Male and female broods were placed in separate tanks for about 24 h with a gentle shower of water for inducing the breeding condition. Both male and female fishes were artificially induced by intra-muscular injection with HCG hormone with a dose of 500 and 3 333 IU/kg body weight, respectively. After injection, the male and female broods were placed in separate spawning tanks. After 12 h of HCG hormone administration, when ovulation seemed to be complete, the broods were taken out from the spawning tank and placed in a blanket for stripping. Female fishes were stripped by gentle pressure on the abdomen to collect the eggs in a plastic bowl. For milt collection, testes were cut into small pieces using a scalpel and mixed with small amount of water, which was then added to the eggs placed in the plastic bowl. Milt and eggs were mixed thoroughly by using clean and a soft poultry feather until fertilization. Immediately after fertilization, the eggs were released into a previously prepared experimental aquarium.

2.3 Experimental setup, sampling and microscopic observation

The stock solution was prepared by dissolving a measured amount of sumithion in distilled water. Two concentrations of sumithion (150 and 250 μg/L) solutions were prepared in two separate aquaria each with 40 L water. The control group was exposed to dechlorinated tap water. Each treatment and control was set up in three replicates.

Twenty-four hours after fertilization, when hatching of eggs occurred, the larvae samples were collected at 12- and 24-h interval until yolk sac absorption to observe the toxicity of sumithion. Five larvae from each treatment were collected in distilled water for immediate microscopic observation

Developmental stages of stinging catfish treated with sumithion as well as a control batch were observed using a digital microscope (Olympus CX 41). Their images were photographed using a microscope camera (Magnus Analytics, Model- MIPS) connected between the microscope and a computer. The developmental stages of stinging catfish were determined according to Puvaneswari et al.(2009).

2.4 Collection of sumithion-affected deformed adult fish

As a part of this experiment, the effect of sumithion in the larval rearing pond was investigated during the adulthood of stinging catfish. Adult fish were collected from a local farm (Sarnalata Agro Fisheries Ltd., Fulbaria, Mymensingh), where the ponds were treated with sumithion before larval rearing.

2.5 Statistical analysis

The data on mortality rate presented in this study are the average of three replicates±standard deviation (SD). Data were analyzed by ANOVA followed by Tukey's post hoc test to assess statistically significant differences among the different treatments. Statistical significance was set at P<0.05. Statistical analysis was performed using SPSS Version 14.0 for Windows (SPSS Inc., Chicago, IL).

3 RESULT 3.1 Toxicity of sumithion on larval stages of stinging catfish

In the present study, larval malformation induced by sumithion was recorded in the treatment groups. Several malformations of larvae characterized by irregular head shape, lordosis, yolk sac edema, body arcuation, tissue ulceration, etc., resulted due to exposure in sumithion (Fig. 1a -h).

Figure 1 Malformations (irregular head shape, lordosis, yolk sac edema, body arcuation, tissue ulceration etc.) induced by sumithion a. anterior part of 6 h hatchling from control treatment; b. posterior part of 6 h hatchling from control treatment; c. irregular head shape and lordosis at 6 h after hatching exposed to 150 μg/L; d. yolk sac edema and body arcuation at 6 h after hatching exposed to 250 μg/L; e. notochord abnormality (body curvature) lordosis and edema at 24 h after hatching exposed to 150 μg/L; f. deformed notochord and edema at 36 h after hatching exposed to 150 μg/L; g. deformed posterior part of body at 36 h after hatching exposed to 250 μg/L; h. tissue ulceration at 84 h after hatching exposed to 150 μg/L.
3.2 Mortality rates of stinging catfish larvae exposed to sumithion

Mortality rates (%) of larvae exposed to sumithion concentrations were estimated at 152 h after exposure and 120 h after fertilization. The mortality rates were 32.0%±2.4%, 57.8%±4.1% and 62.67%±7.6% in control, 150 μg/L and 250 μg/L of sumithion, respectively (Fig. 2).

Figure 2 Mortality rate (%) of larvae exposed to 0 μg/L (control), 150 μg/L and 250 μg/L concentrations of sumithion Values accompanied by different letters are statistically different (P<0.05).
3.3 Deformity in adult stinging catfish in sumithiontreated ponds

We found that 30% of the total adult stinging catfish populations were deformed in ponds treated with sumithion during larval rearing. Different types of physical deformities (mainly deformed notochord) were recorded in adult stinging catfish (Fig. 3).

Figure 3 Deformities induced by sumithion in adults a. normal (non-deformed) adult from the sumithion- treated pond; b-d. deformed adults from the sumithion-treated ponds.
4 DISCUSSION

In the present study, it has been found that sumithion has significant effects on developmental stages of stinging catfish. Developmental abnormalities of larvae, such as irregular head shape, lordosis, yolk sac edema, body arcuation and tissue ulceration were found due to exposure to sumithion. Generally, yolk sac edema has been recognized to be the feature of vertebrate embryo/larvae-toxicity (Hamm and Hinton, 2000). Edema was observed in zebrafish exposed to 2, 3, 7, 8-tetrachlorodibenzo- p - dioxin (Henry et al., 1997) and in medaka fish exposed to thiobencarb (Villalobos et al., 2000). There is some evidence of spinal deformity in medaka exposed to the organophosphorus pesticides diazinon (Hamm and Hinton, 2000) and fenitrothion (Hiraoka, 1989). Several abnormalities, including a wavy notochord, a curving trunk, somite defects, shorter trunk axis, a larger yolk sac, a shorter yolk sac extension and less melanin pigmentation were observed in zebrafish larvae exposed to cartap (Zhou et al., 2009) and bifenthrin (Jin et al., 2009). Interestingly, different types of physical deformities (mostly deformed notochords) were observed in adult stinging catfish collected from ponds treated with sumithion during larval rearing (Fig. 3), indicating long-term effects of sumithion on this fish. It has been reported that the high toxicity of pesticides is greatly related to the nervous system (Oh et al., 1991). Therefore, deformities of stinging catfish larvae in the present study might be due to malfunctioning of the central nervous system by sumithion toxicity.

The mortality rates (%) of stinging catfish larvae exposed to sumithion increased with increasing concentrations of sumithion. This finding is strongly supported by earlier findings in salmon embryos (Lower and Moore, 2003) and common carp larvae (Aydin and Koprucu, 2005) exposed to organophosphorus pesticide diazinon. Takimoto et al.(1984) also reported different degrees of mortality in Oryzias latipes embryos exposed to sub-lethal concentrations of the organophosphate fenitrothion. It was noticed that movements in the larvae exposed to sumithion decreased, indicating the adverse effects of sumithion on activities of stinging catfish larvae. A variety of neurotoxic contaminants, ranging from pesticides to metals, may cause abnormal swimming behavior or conciliation swimming ability of fish and other aquatic animals (Christensen et al., 2005 ; Werner and Moran, 2008 ; Jin et al., 2009). A decrease in swimming speed after exposure to organophosphates and carbamates has been noticed in rainbow trout (Brewer et al., 2001), seabass (Almeida et al., 2010) and tilapia (Pessoa et al., 2011). Hence, it is clear that increased mortality and slow movements of stinging catfish larvae in the present study were due to sumithion toxicity.

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