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  • Study Of Toxic Effect Of Cypermethrin EC (10%) On Slender Rasbora, Rasbora Daniconius (Hamilton, 1882)

  • 1Department of Zoology, Sreekrishna College, Guruvayur, Kerala, India
    2University of Calicut, Thenhipalam, Kerala, India
    3Amritha Vishwa Vidyapeetham, Kollam, Kerala , India
     

Abstract

Cypermethrin, a type II synthetic pyrethroid insecticide is commonly utilised in agricultural and domestic applications. The excessive use of this can infiltrate freshwater ecosystems, impacting non-target species such as the slender rasbora Rasbora daniconius (Hamilton, 1882). This study evaluated experimentally the lethal and sublethal effects of cypermethrin. The LC50 value was determined as 3 ppm (96 hours) using probit analysis. The effects of a sublethal dose of cypermethrin 3 ppm after 10 and 15 days post-exposure showed significant protein depletion in the fish. Behavioural changes were also noted after treating the fish at sub-lethal concentrations.

Keywords

Cypermethrin, LD50, Sublethal dose, Protein Concentration, Rasbora daniconius

Introduction

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Cypermethrin is a pyrethroid insecticide widely used in homes, agriculture, and industrial settings for controlling various insect pests [1](Kakko et al., 2003). Cypermethrin exhibited stability against photochemical and microbial degradation and had very low toxicity to humans and animals. This insecticide has become more widely used than organochlorine, organophosphorus, and carbamate insecticides [2] (Coats et al., 1989)..  It is significantly utilized in both agricultural and domestic contexts. Currently, approximately 30% of pesticides used globally are synthetic pyrethroids [3] (Oda and Maddawy, 2012). The excessive and immoderate use of cypermethrin can lead to runoff into natural water bodies, posing significant risks to non-target aquatic organisms. Behavioural, haematological, and histological changes are critical effects of pesticide exposure at higher concentrations in fish. The toxic effects of cypermethrin pose a significant threat to aquatic ecosystems.

The slender rasbora, Rasbora daniconius (Hamilton, 1882), is one of the common fish species inhabiting the freshwater environments of paddy fields. The study investigates the acute toxicity of the pyrethroid cypermethrin on slender rasbora collected from paddy wetlands in the Pannissery area near Sreekrishna College, Guruvayur. Toxicity tests revealed that LC50 of cypermethrin for the fish to be 4 ppm using probit analysis.  Toxicity study at sublethal  concentration  of 2ppm for 10 days  and 15 days post-exposure to slender rasbora demonstrates the significant behavioural changes and a substantial decrease in protein content in the treated fish. The comprehensive data of protein concentration depletion due to pesticide toxicity in this species underscore the need for responsible pest management practices in both domestic and agricultural settings.

SUBJECTS AND METHODS:

Experimental set up (Test specimen, Collection method  and Insecticide formulation)

The fish used for the experiments were collected from the paddy wetlands in Pannissery near Sreekrishna College . They were collected using cast and drag nets. Twenty healthy freshwater fish, Rasbora daniconius, with an average length of 5-8 cm and a weight of 20 ± 2 g, were used for the study. The fish were cleaned using a 0.5% potassium permanganate solution to remove any contaminants. The fish were acclimatised for 7 days in an aquarium of 50 L capacity in dechlorinated aerated water at room temperature (26 ± 2 °C).

Study of Lethal dose - LD50

Cypermethrin (10% EC), manufactured by National Pesticides and Chemical, Mumbai, was purchased from the agricultural market in Thrissur, Kerala, India. A stock solution of cypermethrin was prepared in distilled water for experiment. Test series concentrations of 1, 2, 3, and 4 μg/l were prepared by serial dilution of the stock solution [4] (Neglur et al., 2020). The fish were  in pesticide-free water served as  control. The treated water in the aquaria was changed after 24 hours, and freshly prepared concentrations of cypermethrin were used for the experiment. The test was conducted for 96 hours, during which dead fish were removed. Probit analysis was employed to determine the median lethal concentration [5] (Finney, 1971).

Sublethal toxicity study

For the acute toxicity study, the fish were maintained in aquaria up to 10 days and  15 days. For Control experiment the aquarium  was filled with dechlorinated, aerated  water at room temperature (26 ± 2 °C).  For experiment the aquarium was filled with water containing 3 μg/l cypermethrin concentration. The treated water was changed every 24 hours  and a fresh concentration of 3 μg/l was maintained throughout the period. The behaviour of the fish, i.e., swimming, opercular movement, etc., was recorded after 96 hours of observation. Protein concentration was assessed after  10 and 15 days post-exposure using a standard procedure [6]Lowri (1957).

Protein Concentration study

The fish were sacrificed periodically, and whole muscle tissues from both treated and untreated fish were homogenised using a mortar and pestle. Protein concentration in fish whole-body homogenates is typically determined by extracting protein using buffer (e.g., 0.1M KCl buffer, centrifuging to obtain a clear supernatant and analysing it with colorimetric method  of against a BSA standard curve, The  process involves precise weighing, homogenization at 4?C. Data obtained from both the control and experimental groups were analysed and presented graphically.

RESULTS:

Lethal study 

The LC50 of cypermethrin was determined using Probit analysis.  Mortality was observed for 24 hours , 48 hours and 96 hours of study. The observation of experiment is presented in Table 1. All fishes died when treated with  4 μg  cypermethrin after 96 hours of observations. The LD50 of cypermethrin is 3 μg concentration after 96 hours of observations.

Table 1. Observation (96 hours) of Mortality in Rasbora daniconius (n=10)  when treated with    different concentrations of (EC 10%) Cypermethrin

Experiment

Cypermethrin Concentration

Mortality Observed after 24 hours

Mortality Observed after 48 hours

Mortality Observed after 96 hours

1

1 μg/l

0

0

0

2

2  μg/l

0

0

2 died

3

3 μg/l

0

0

3 died

4

4  μg/l

0

0

10 died

Sublethal toxicity study

Exposure of slender rasbora to a sublethal dose of cypermethrin 3 μg/l for 10 days and 15 days resulted in significant  behavioural changes and biochemical alterations, particularly a depletion of protein concentration. This depletion indicates dose-response pressure or metabolic disturbances following pesticide exposure.

Behavioural Studty

Behavioural modifications were also observed along with the biochemical changes. The observations of behvariour  were presented in Table 4. The behaviours changes were recorded for  hyperactivity, loss of balance, rate of swimming, opercular activity and surfacing activity and  air gasping activity. Such changes contribute to the overall compromise of the fish's health and survival.

Table 2. Impact of Cypermethrin (10% EC) on the behaviour pattern of Rasbora daniconius

Parameter’s

Control

Sublethal dose 3 μg/l

Sublethal concentration

96 hours (3 days)

5  days

Hyperactivity

−

++

+++

Loss of balance

−

+

++

Rate of swimming

−

++

+++

Opercular activity

−

++

+++

Surfacing Activity

−

++

+++

Air gasping activity

−

+

++
  • indicate normal behaviour, + indicate increasing behaviour

Protein depleti

Statistical analysis of the protein depletion after 10 days and 15 days exposure to the slender rasbora was done using ‘t’ value. After 10 days protein concentration was depleted to 83% in test specimen while in 15 days it is 88.5 %. This strongly suggests that the toxicant has a severe adverse biochemical effect, likely due to: Protein degradation, Metabolic stress and Impaired protein synthesis.

Statistiical analysis (Table 3)

Table 3.  Changes in the Protein Concentration when exposure to different periods of sublethal toxicity 10 days and 15 days

Days of exposure

Protein Concentration

 

Control

Treated

10 days

3.08+.05

0.50+.03

% Changes

 

83.8

‘t’ value

75.9

15 days

3.74±.07

0.113±.005

% Changes

 

97%

‘t’ value

88.5

DISCUSSION :

The present study determines the LC50 value of cypermethrin to the Slender Rasbora, Rasbora daniconius is 3 μg/l. [7]Adhikari et al. (2005) reported the toxicity of cypermethrin in laboratory tests.  In aquatic invertebrates LD50  toxicity ranging from 0.01 to 5 μg/l.  In Table 4 the study of the effects of cypermethrin on various freshwater fishes were presented. 

In Table 4, the study of the effects of cypermethrin on various freshwater fish species was presented. From the data in Table 4, it is evident that cypermethrin impacts freshwater fish at both genetic and physiological levels. The table also discusses behavioural, biochemical, and histopathological changes in the affected fish. This indicates that cypermethrin exhibits high toxicity even at low concentrations. To better understand the toxic effects of cypermethrin on fish, it is recommended to examine behavioural changes in conjunction with biochemical parameters, such as protein concentration, and other metabolic functions.

Table 4.The toxic effects of Cypermethrin invarious fish species with reference

SI.No.

Model Organism

Changes Observed

Reference

1

Channa punctata

Genotoxicity and Oxidative stress in fish erythrocytes

Ansari and Kumar (1988) [8]

3

Oncorhynchus mykiss trout

Biochemical parameters

Atamanalp et al (2002) [9]

4

Rohu (Labeo rohita)

Haemato-biochemical alterations

Das and Mukherjee (2003) [10]

5

Rainbow Trout

Paramount effects on different haematological parameters

Cakmak(2003) [11]

6

Labeo rohita

Haematological changes

Adhikari et al (2004)

[12]

7

Cirrhinus mrigala

Protease activity and free amino acid levels have significantly increased

Prashanth and David (2006) [13]

8

Cirrhinus mrigala

significant changes in the ammonia, urea, and glutamine levels

Prashanth (2007) [14]

11

Clarias gariepinus

Histopathological effects

Ayoola and Ajani (2008) [15]

12

Heteropneustes fossilis

Disturbance to spermatogenic cells and follicular wall

Carriquiriborde et al. (2009) [16]

13

Clarias batrachus

Biochemical alterations

Begum G (2009) [17]

15

Labeo rohita

Hyper excitability, erratic swimming, loss of equilibrium and sinking to the bottom

Marigoudar et. al. (2009) [18]

16

Channa punctata

Cytogenetic and oxidative stress

Ansari et al (2011) [19]

17

Zebrafish (Danio rerio) embryos

Developmental toxicity

Shi et al., (2011) [20]

18

Catla catla

Alterations in anti-oxidative enzymes

Akinrotimi et al (2012) [21]

19

Clarias gariepinus

Haematological changes

Akinrotimi et al (2012) [21]

20

Catla catla

Biochemical and Haematological parameter changes

Kannan et al., (2014)

[22]

21

Clarius batrachus

Marked alterations in the level of LPO

Kumar et al.,(2014)

[23]

22

Jenynsia multidentate

Changes in acetylcholinesterase activity and erratic swimming

Bonansea et al., (2016) [24]

23

Anabas testudineus

Haematological changes

Velmurugan et al (2016) [25]

24

Danio rerio

Oedema of and yolk sac and pericardium

Xu C et al (2018) [26]

25

Heteropneustes fossils

Glutamate dehydrogenase (GDH) enzyme activity was elevated in the liver and gill tis- sues.

Sulthana RN (2021)

[26 [27]

26

Channa punctata

Gill toxicity

Dubey and Sharma (2022) [28]

27

Channa punctata

Alterations in the haematological parameter

Ghosh et al. (2022) [29]

28

Catla catla

DNA damage

Sharma and Jindal (2022) [30]

29

Mystus cavasius

Negative impact on the haemato-biochemical parameters and female gonad

Uddin et al (2022) [31]

30

Aplocheilus lineatus

Behavioural and Protein constituents

Sai Krishna et al (2024) [32]

Cypermethrin exposure causes abnormal swimming patterns, reduced opercular activity, and altered air-gasping behaviour in fish. Fish subjected to pesticide stress sometimes exhibit symptoms of dullness, loss of equilibrium, loss of appetite, and erratic swimming [33] (Kavitha and Rao, 2007). [34]Ural and Koprucu (2006) also reported that fish began breathing air at the surface, swimming on the water's surface, and displaying altered swimming behavior from the first day when exposed to sublethal concentrations.  [35]Kaviraj and Gupta (2014) observed that cypermethrin induces oxidative stress in fish, initially leading to frequent surfacing, increased opercular movement, and faster swimming activity. However, within 24 hours of treatment, the fish gradually displayed lethargy in their movements.

The acute toxicity of this pesticide at sublethal concentrations was examined through observation of behavioural and biochemical changes. After 10 and 15 days of cypermethrin exposure, protein concentrations in the muscle tissue of the fish decreased by more than 80%. The sublethal effects of cypermethrin on protein metabolism suggest that the reduction in protein content in the experimental fish may be attributed to several factors. The effect of cypermethrin on protein concentration has been studied by[36] Sambasiva Rao (1999), [37]Li et al. (2005) and [38]Thenmozhi et al. (2011). These reductions in protein levels may be due to increased energy demands and the breakdown of biomolecules as the fish cope with pesticide-induced metabolic stress [39] (Kumari et al., 2024).

CONCLUSION:

The effects of cypermethrin on the slender rasbora, Rasbora daniconius cause acute toxicity in the fish and lead to changes in behavioral and protein aspects. There was a higher degree of decrease in the level of protein content in the whole-body extract. The present research  highlights the harmful effect caused by cypermethrin to the fish populations and emphasizes the need for careful regulation and monitoring to adjudicate its  effective use on aquatic ecosystem.

ACKNOWLDGEMENTS:

Authors hereby acknowledge the Department of Zoology, Sreekrishna College, Guruvayur, for the lab facilities provided.

Author’s statements

 Authors      clearly states that there are no potential conflicts of interest in the article. No funding agency resources are available for this study. While handling the specimens, ethical procedures and standards are maintained.

Authors’ Contributions-

Brinesh R prepared the manuscript, conceived the study, Dilsha Parvin and Devika K researched the literature, and contributed in the lab work.

Ethical Issues

There were no ethical issues and concerns in the experiment and result in my original research paper.

Conflict of Interest

Author declares that there is no conflict of interest while doing the experiment and preparing the manuscript.

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  46. Prashanth M S, David M: Changes in nitrogen metabolism of the freshwater fish Cirrhinusm kigala following exposure to Cypermethrin. Jour of Basic Clin Physiol and Pharmac , 2006; 17,63–70.
  47. Prashanth M S :  Cypermethrin-induced protein metabolism in the freshwater fish Cirrhinus mrigala (Hamaliton). Journal of  Bas Clini Physiol and Pharmacol, 2007; 18:49–64.P arma MJ,
  48. Borges, et. al. : Changes in hematological and serum biochemical values in jundiá Rhamdiaquelenduetosub-lethal toxicity of cypermethrin.Chemosphere, 2007; 69, 920–926.
  49. Ayoola SO, Ajani E K : Histopathological effect of cypermethrin on juvenile African catfish (Clarias gariepinus). World Journal of Biol Res , 2008;1:1–14.
  50. Carriquiriborde P, Díaz J, López C, Ronco A E, Somoza GM : Effects of cypermethrin chronic exposure and water temperature on survival, growth, sex differentiation, and gonadal developmental stages of Odontes thesbonariensis (Teleostei). Chemosphere, 2009;76, 374–380. https://doi.org/10.1016/j.chemosphere.2009.03.039.
  51. Marigoudar, SR, Ahmed, RN, David, M :  Cypermethrin-induced in vivo inhibition of the acetyl cholin esterase activity in functionally different tissues of the freshwater teleost, Labeo rohita (Hamilton). Toxicol Environ Chemistry, 2009; 91, 1175–1182.
  52. Ansari RA, Rahman S, Kaur M, Anjum S, Raisuddin S : Invivo cytogenetic and oxidative stress-inducing effects of cypermethrin in freshwater fish,   Channa punctata Bloch, 2011; Ecotoxicology, Environment Safety, 74,150–156.
  53. Shi X, Gu A, Ji G, Li Y, Di J, Jin J, Wang X  Developmental toxicity of cypermethrin in embryo-larval stages of zebrafish. Chemosphere, 2011; 85:1010–1016. https://doi.org/10.1016/j.chemosphere.2011.07.024.
  54. Akinrotimi OA, Gabriel UU, Ariweriokuma SV:  Haematotoxicity of cypermethrin to African catfish Clarias gariepinus under laboratory conditions. Environment Science, 2012; 1,13–19.
  55. Kannan M, Muthusamy P, Venkatachalam U : Response of synthetic pyrethroid cypermethrin (10% EC) induced stress in biochemical and hematological parameters of Indian major carp Catla catla (Hamilton,1822).W Jour of  Pharma Res, 2014; 3,1976–1996.
  56. KumarA, Sharma B, Pandey RS:   λ-Cyhalothrin and cypermethrin induce stress in the freshwater muddy fish, Clarias batrchus.  Toxico. Environ. Chem,2014; 96,136–149.
  57. Bonansea, RI, Wunderlin DA, Amé MV:  Behavioral swimming effects and acetylcholinesterase activity changes in Jenynsia multidentata exposed to chlorpyrifos and cypermethrin individually  and in  mixtures . Ecotoxicol and Enviro Saft,2016; 129:311–319

Reference

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  2. Oda S S , El-Maddawy ZK : Protective effect of vitamin E and selenium combination on deltamethrin-induced reproductive toxicity in male rats. Experimental Toxicololgy and Pathology, 2012;64,813–815.
  3. Ullah, SMZ AA : V Sci and Univ, 2015; Nexus_ MH20141213011243, Nexus academic publishers.com, 3,40–57.
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  49. Ayoola SO, Ajani E K : Histopathological effect of cypermethrin on juvenile African catfish (Clarias gariepinus). World Journal of Biol Res , 2008;1:1–14.
  50. Carriquiriborde P, Díaz J, López C, Ronco A E, Somoza GM : Effects of cypermethrin chronic exposure and water temperature on survival, growth, sex differentiation, and gonadal developmental stages of Odontes thesbonariensis (Teleostei). Chemosphere, 2009;76, 374–380. https://doi.org/10.1016/j.chemosphere.2009.03.039.
  51. Marigoudar, SR, Ahmed, RN, David, M :  Cypermethrin-induced in vivo inhibition of the acetyl cholin esterase activity in functionally different tissues of the freshwater teleost, Labeo rohita (Hamilton). Toxicol Environ Chemistry, 2009; 91, 1175–1182.
  52. Ansari RA, Rahman S, Kaur M, Anjum S, Raisuddin S : Invivo cytogenetic and oxidative stress-inducing effects of cypermethrin in freshwater fish,   Channa punctata Bloch, 2011; Ecotoxicology, Environment Safety, 74,150–156.
  53. Shi X, Gu A, Ji G, Li Y, Di J, Jin J, Wang X  Developmental toxicity of cypermethrin in embryo-larval stages of zebrafish. Chemosphere, 2011; 85:1010–1016. https://doi.org/10.1016/j.chemosphere.2011.07.024.
  54. Akinrotimi OA, Gabriel UU, Ariweriokuma SV:  Haematotoxicity of cypermethrin to African catfish Clarias gariepinus under laboratory conditions. Environment Science, 2012; 1,13–19.
  55. Kannan M, Muthusamy P, Venkatachalam U : Response of synthetic pyrethroid cypermethrin (10% EC) induced stress in biochemical and hematological parameters of Indian major carp Catla catla (Hamilton,1822).W Jour of  Pharma Res, 2014; 3,1976–1996.
  56. KumarA, Sharma B, Pandey RS:   λ-Cyhalothrin and cypermethrin induce stress in the freshwater muddy fish, Clarias batrchus.  Toxico. Environ. Chem,2014; 96,136–149.
  57. Bonansea, RI, Wunderlin DA, Amé MV:  Behavioral swimming effects and acetylcholinesterase activity changes in Jenynsia multidentata exposed to chlorpyrifos and cypermethrin individually  and in  mixtures . Ecotoxicol and Enviro Saft,2016; 129:311–319

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R. Brinesh
Corresponding author

Department of Zoology, Sreekrishna College, Guruvayur, Kerala, India

Photo
Dilsha Parvin
Co-author

University of Calicut, Thenhipalam, Kerala, India

Photo
K. Devika
Co-author

Amritha Vishwa Vidyapeetham, Kollam, Kerala , India

R. Brinesh, Dilsha Parvin, K. Devika, Study Of Toxic Effect Of Cypermethrin EC (10%) On Slender Rasbora, Rasbora Daniconius (Hamilton, 1882), Int. J. of Pharm. Sci., 2026, Vol 4, Issue 8, 880-889. https://doi.org/10.5281/zenodo.21812758

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