M. suseela et al Adv.Res.J.Bios.Mol.Tech.

, 2015,Vol I:Issue 1; (5-11)

Available on online www.arjbsmt.com

Advance Research Journal of Biological Sciences and Molecular Techniques

---------------------------------------------------------------------------------------------------------------------

Adv.Res.J.Bios.Mol.Tech., 2015,Vol I:Issue 1; (5-11) (ISSN:2395-7689)

---------------------------------------------------------------------------------------------------------------------

ESTIMATION OF TOXICITY EVALUATION (LD50) OF LAMBDA CYHALOTHRIN, A

SYNTHETIC PESTICIDE ON ALBINO MICE

*M.Suseela1, K.Gokul1, K. Jayantha Rao2 and P. Jacob Doss2

1
Department of Zoology, Vikrama Simhapuri University, Nellore
1
Department of Zoology, Rayalaseema University, Kurnool
2
Department of Zoology, Sri Venkateswara University, Tirupati
* Corresponding author; Email:drsuseelameesala@gmail.com

ABSTRACT

Pesticides are designed to control pests. They all are toxic to some level; otherwise they
would not kill pests. They can also be toxic to non-target organisms such as plants, animals or
humans. Exposure to a sufficient amount of almost any pesticide can affect a personeither
through illness, eye exposure or skin sensitivity. Excess of any compound will be harmful to life
and consider leader toxicity studies Pesticides are use for welfare of human beings but by the
time, they will challenge us by showing their toxicity. Pesticide pollution brings about sudden &
drastic changes. The biological effects of pesticides and their mode of action are obtained from
toxicity studies. The present study aim to find out the short term exposure effect of Lambda
cyhalothrin on Albino mice with various doses 8,12,16,20,24,28,36,40,48,52 mg/kg bw
respectively. By the study of toxicity evaluation by the Probit Analysis method of Finney, LD50
of Lambda cyhalothrin has been found to be 24mg/kg bw with the exposure period of 48hrs.

Key Words: Lambda cyhalothrin, LD50, Toxicity Evaluation, Probit Analysis method

---------------------------------------------------------------------------------------------------------------------

INTRODUCTION

All substances are poisons; there is none which is not a poison. The right dose differentiates
a poison from a remedy.
- Paracelsus.

5
M. suseela et al Adv.Res.J.Bios.Mol.Tech., 2015,Vol I:Issue 1; (5-11)
In the thirst of modernization and industrialization man has contributed pollution to the life and
ecology of plants, animals and microbes. Increased demand for food and fiber has lead to the
chemicalization of agriculture and we have reached on such a stage, that modern agriculture is
dependent on high yielding varieties, which can only be grown under the influence of fertilizers
and pesticides (Chauhan, 2009). Among the various pollutants, chemical pollution of the
environment occupies a pivotal place. Due to lack of highly selective pesticidal action, threat to
non-target forms imposed by various pesticidal uses in agriculture needs to be monitored and this
needs thorough study of individual pesticidal formulations on the biology of non target
organisms (Jacob doss et al, 2006). Pest induced crop loss is on the rise despite increasing usage
of pesticides (Bikramjit Sinha and Indranil Biswas, 2009).

Excess of any compound will be harmful to life and consider leader toxicity studies
Pesticides are use for welfare of human beings but by the time, they will challenge us by
showing their toxicity. Pesticide pollution brings about sudden & drastic changes. Pesticides are
toxic compounds to all living organisms however effects vary with species to species but
excessive use of these pesticides create many problems to all of us. These days, synthetic
chemical pesticides are in practice because of their active and best results. But their excessive
use causes serious damage to eco system-terrestrial as well as aquatic and consequently the flora
and fauna of surrounding (Paliwal et al., 2009). Now a days synthetic pyrethroids have become
an economically and environment friendly group of insecticides. The persistence and continuous
application of these synthetic pyrethroids may create a problem directly or indirectly in the
higher tropical level of the ecosystem.

The biological effects of pesticides and their mode of action are obtained from toxicity
studies (Ambika and Selvisabhanayakam, 2012). Toxicity refer to the ability of a poison to
produce adverse effects. Toxicity is usually divided into 2 types, acute or chronic, based on the
number of exposures to a poison and the time it takes for toxic symptoms to develop. Acute
toxicity is due to short-term exposure and happens within a relatively short period of time,
whereas chronic exposure is due to repeated or long-term exposure and happens over a longer
period. The acute toxicity of a chemical refers to its ability to do systemic damage as a result of a
one-time exposure to relatively large amounts of chemical. The commonly used term to describe
acute toxicity is LD50. LD means Lethal Dose (deadly amount) and the subscript 50 means that
the dose was acutely lethal for 50 percent of the animals to whom the chemical was administered
under controlled laboratory conditions.

Acute toxicity is measured as the amount or concentration of a toxicant. LD50 and LC50
values are useful in comparing the toxicities of different active ingredients and different
formulations containing the same active ingredient. The lower the LD50 or LC50 of a pesticide
product, the greater its toxicity to humans and animals. Pesticides with a high LD50 are the least
toxic to humans if used according to the directions on the product label. The application of
LD50value has gained wide acceptance among toxicologists and if generally the most reliable test
accessing potential hazards of Aquatic and terrestial life (Brugngs and Mount, 1978).

The LD50 values are differ from species to species for the same toxicants due to the mode
of action and response of the animals. The result is expressed as the lethal dose (LD) in case of
terrestrial and lethal concentration in case of aquatic organisms. Since some members of
population may prove to be excessively susceptible and others may prove to be very resistant to

6
M. suseela et al Adv.Res.J.Bios.Mol.Tech., 2015,Vol I:Issue 1; (5-11)
the dose or the concentration of the toxicant that affects 50% of the population under
consideration is expressed as LD50 or LC50 values, which is statistically calculated on the basis of
the observed percentage of mortality at different concentrations of the pesticides (Venkata
Ratnamma and Nagaraju, 2013). The toxicity of pesticide measured in several ways but
generally human toxicity is estimated based on the test results on rats and other animal models
(Jaya Raj et al., 2013). The LD50 values of new drugs are determined by the various route of
administration (intravenous, intraperitonial, subcutaneous and oral). Result of LD50 study may
affect by the some factors which are species, age, sex, amount of food, social environment,
route of exposure (oral, dermal, inhalation) and physical environment such as temperature and
humidity (Deora paramveer et al., 2010). In view of this, an attempt has been made in the
present study to find out the LD50 values for the albino mice when exposed to a synthetic
pyrethroid Lambda cyhalothrin.

MATERIAL AND METHODS

Chemical Substance and Experimental Animal

Lambda cyhalothrin, a synthetic pyrethroid has been considered for this toxicology study.
The toxicological studies have been done on albino mice which are closely related to human
beings and easy to handle in laboratory. Albino mice of 305g were selected from an inbred
colony for the experimentation. The mice were maintained at 255c temperature. They were
provided food pellets and clean water for survival.

Determination of LD50

The LD50 was determined by log-dose/probit regression live method (Finney, 1971). Ten
sets containing 10 mice each were taken and ten serially diluted doses were given orally with the
help of gavage tube. The doses were 8,12,16,20,24,28,36,40,48,52 mg/kg bw for Lambda
cyhalothrin. The mice were observed for 48hrs and then mortality was noted for further
calculation. A graph has been plotted between empirical probit and log dose and then LD50 has
been calculated with the help of regression live and computerized calculation.

Results and Discussions

In the present investigation the mortality rates of mice were determined by giving the
Lambda cyhalothrin orally. Significantly to identify the lethality of Lambda cyhalothrin in
different concentrations/doses ranging from 8 mg/kg body weight were used. The mortality
showed in the percentage at different doses after introducing Lambda cyhalothrin to the
mammalian model albino mice.

The data was computed according to Probit Analysis Method (Finney, 1971) and LD50
value was determined. The animals are exposed to different concentrations of Lambda
cyhalothrin, showed no mortality up to 8 mg/kg body weight, 10 percent mortality at 12 mg/kg
bw, 20 percent mortality at 16 mg/kg bw, 30 percent mortality at 20 mg/kg bw, 50 percent
mortality at 24 mg/kg bw, 70 percent mortality at 28 mg/kg bw, 80 percent mortality at 36 mg/kg
bw,90 percent mortality at 40 mg/kg bw and 100 percent mortality at 48 mg/kg bw and 52 mg/kg
bw observed (Table 1.1).

7
M. suseela et al Adv.Res.J.Bios.Mol.Tech., 2015,Vol I:Issue 1; (5-11)

The computation of Percent Mortality against different Log concentrations of the

pesticide yielded a typical sigmoid curve (Figure 1.1). The LD50 value obtained from the
sigmoid curve is 24 mg/kg bw for 48 hours. The Probit Mortality of the albino mice were
calculated from Percent Mortality. When the Probit Morality was plotted against Log
concentrations of the pesticide, a straight line was obtained (Figure 1.2). The LD50value
obtained from this straight line graph is 24 mg/kg bw. LD50 of Lambda cyhalothrin was noted as
24mg/kg bw.

The graphical representation of Percent Mortality versus Log concentration and Probit
Mortality versus Log concentration of Lambda cyhalothrin (Table 1.1) showed a typical sigmoid
curve (Figure 1.1) and a straight line (Figure 1.2) respectively which are in agreement with the
principle of Probit Analysis (Finney, 1971).

In the present investigation, the obtained LD50 value is 24 mg/kg bw. This value is in
agreement with LD50 value reported previously by Mosbah Rachid et al., (2010). Accordingly
to these references the rats LD50 value was found to be 18-32 mg/kg bw.

Every pesticide may vary greatly in its toxicity and persistence. Since the evaluation of
toxicity of a test chemical is a sensitive phenomenon, which can be influenced by several factors
such as size (Jayantha Rao, 1982), nutritional status (Pal and Kushwah, 1981; Das and Garg,
1981), species specificity (Gouda et al., 1981; Jacob et al., 2006; Janardhan et al., 1987),
animal weight (Pickering et al., 1962), its developmental stage, time of exposure and
temperature (Macek et al., 1969). Thus, various factors influence the LD50values. These factors
are manifold and dependent upon the given set of experimental conditions (Russell and
Overstreet, 1987). The LD50 values can also significantly vary between animals of the same
basic strain obtained from different suppliers (Russell and Overstreet, 1987), according to the
purity of the chemical (HO et al., 1983) and the sex difference (Overstreet et al., 1979). It has
also been observed that the LD50 differs between water deprived and non-deprived animals
(Russell et al., 1986).

Various factors influence the LD50 values. These factors are manifold and dependent upon
the given set of experimental conditions (Russell and Overstreet, 1987). Various factors like
vehicle, isomers of Lambda cyhalothrin, percentage of corn oil and the environmental conditions
influences the lethal dosage of Lambda cyhalothrin. The LD50 values significantly vary from
species to species (Pickering et al., 1962), capacity of the species to tolerate the pesticide
(Chambers and Yarbrough, 1974), the LD50 values can also significantly vary between animals
of the same basic strain obtained from different suppliers (Russell and Overstreet, 1987),
according to the purity of the chemical (Ho and Hoskins, 1986) and to the sex differences
(Overstreet et al., 1979). The differences in LD50 value between water-deprived and non-
deprived were also observed. So the differences of LD50 obtained in the present investigation
from the reported LD50 values by other investigators may be due to one (or) more factors as
listed above.

8
M. suseela et al Adv.Res.J.Bios.Mol.Tech., 2015,Vol I:Issue 1; (5-11)
Table 1.1: Mortality of albino mice exposed to different concentrations of Lambda
cyhalothrin at 48 hours. (Mortality was expressed both in percent and probit kill)

S.No. Concentration Log Number of animals Percent Probit

mg/kg body concentration Kill Kill
weight Exposed Dead
1. 8 0.903 10 0 0 -
2. 12 1.079 10 1 10 3.71
3. 16 1.204 10 2 20 4.16
4. 20 1.301 10 3 30 4.47
5. 24 1.380 10 5 50 5.00
6. 28 1.477 10 7 70 5.52
7. 36 1.556 10 8 80 5.84
8. 40 1.602 10 9 90 6.28
9. 48 1.681 10 10 100 8.09
10. 52 1.716 10 10 100 8.09

Fig 1.1: Sigmoid curve showing mortality of albino mice against log concentration of
Lambda cyhalothrin

9
M. suseela et al Adv.Res.J.Bios.Mol.Tech., 2015,Vol I:Issue 1; (5-11)
Fig 1.2: Probit regression line showing mortality of albino mice against log concentration
of Lambda cyhalothrin

References

Ambika A and Selvisabhanayakam (2012). Toxicity studies on the effect of latex on the adult
male insect, Odontopus varicornis (Heteroptera: Pyrrhocoridae). International Journal of
Toxicology and Applied Pharmacology 2012; 2(3): 42-44.
Bikramjit Sinha and Indranil Biswas (2009). Potential of Bio-pesticides in Indian Agriculture
Vis--vis Rural Development. Council of Scientific and Industrial Research (CSIR)
National Institute of science Technology and Development studies (NISTADS).
Brugngs W A and Mount DI (1978). Intoduction to a discussion on the use of aquatic toxicity
test for evaluation of the effects of toxic substances. In estimating the hazard of chemical
substances to aquatic life. ASTMST, (1978) PP 657.
Chambers J.E and J.D. Yarbrough (1974). Parathion and methyl parathion toxicity to
insecticide resistant and susceptible mosquito fish, Gambusia affinis. Bulletin of
Environmental Contamination Toxicology., 11: 315.
Chauhan R.S (2009). Beware of pesticides in food chain. Home> 2009 Issues> August 16,
2009.
Das N and A Garg (1981). Effect of endosulfan in female rat grown on low protein and high
protein cereal diet. Pestic. Biochem. Physiol., 5(1): 90-98.
Doera Paramveer S, Mishra Chanchalk, Mavani Paresh, Asha Rani, Shrivastava B and
Rajesh Kumar Nema (2010). Effective alternative methods of LD50 help to save
number of experimental animals. J.Chem.Pharm.Res., 2010, 2(6); 450-453.
Finney D.J (1971). Probit Analysis, Third Edition, Cambridge University Press, London.303 pp
Gouda R.K, N.K Triphathy and C.C Dass (1981). Toxicity of dimecron, sevin and lindane to
Anabas Scandens and Heteropneustes fossilis. Comp. Physiol. Ecol., 6(3): 170-172.

10
M. suseela et al Adv.Res.J.Bios.Mol.Tech., 2015,Vol I:Issue 1; (5-11)
Ho I.K and B Hoskins (1986). Biochemical and pharmacological aspects of neurotoxicity from
and tolerance to organophosphate cholinesterase inhibitors. In: Hand book of toxicology.
Eds. T.J. Haley, and W.O. Berndt. Hemisphere publishing corp. Washington.
Ho I.K, S.P Sivam and B Hoskins (1983). Acute toxicity of DFP, Tabun, Sarin and Soman in
rats: Lethality in relation to cholinergic and GABAergic enzyme activities. Fed. Proc.,
42: 656 (Abs).
Jacobdoss P, Nagarjuna A, Suhasini K, Savithri Y, Dayanand and Rajeswar Rao M (2006).
Impact of monocrotophos on Albino Rat Neural nitric oxide synthatase activity invivo. J.
Natcon., 18(2): 305-310 (2006).
Janardhan A, A B Rao and P Sisodia (1987). Sub-chronic toxicity of methyl benzimidazole
carbamate in rats. Bulletin of Environmental Contamination and Toxicology.,38(5): 890-
8.
Jaya Raj, Mohineesh, Tiranth D. Dogra, Monika Pahuja and Anupuma Raina (2013).
Determination of Median Lethal dose of combination of Endosulfan and cypermethrin in
Wistar Rats. Toxicology International. Jan-Apr, 2013/Vol-20/Issue1.
Jayantha Rao K (1982). Effect of a systemic pesticide, Phosphomidon on some aspects of
metabolism in fresh water fish, Tilapia mossambica (Peters). Ph.D. Thesis, Sri
Venkateswara University, Tirupati, India.
Macek K.J, C. Hutchinson and O.B. Cope (1969). Bulletin of Environmental and
Contamination Toxicology., 4: 174.
Mosbah Rachid, Boulakoud Mohamed Salab and Yousef Ibrahim Mokhtar (2010). Effects
of Lambda cyhalothrin on haematological parameters and testicular functions in male rat.
Endocrine abstracts (2010), 22 P371.
Overstreet D.H, R.W. Russell, H.C. Helps and M. Messenger (1979). Selective breeding for
sensitivity to the anticholinersterase DFP. Psychopharmacology., 65: 15-20.
Pal A.K and H.S. Kushwah (1981). A. Preliminary study on protective role of protein against
endosulfan exposure. Ind. J. Biophys. Biochem., 8: 4-10.
Paliwal A, Gujar R.K, Sharma H.N (2009). Analysisof liver enzymes in albino rat under stress
of Lambda cyhalothrin and Nuvan toxicity. Toxeminat-1, Feb 22 2009, Biology and
Medicine. Vol. (2): 70-73.
Pickering Q.H, C.S. Henderson and A.E. Lemke (1962). Toxicity of organophosphorous
insecticides to different species of warm water fishes. Trans. A.M. Fish. Soc., 91: 175-
184.
Russell R W, R A Booth, S D Lauretz, C A Smith and D J Jenden, (1986). Behavioral
neurochemical physiological effects of repeated exposures to subsymptomatic levels of
the anticholinesterase, soman. Neurobehavioral. Toxicology and Teratology., 8: 675-685.
Russell R.W and D.H Overstreet (1987). Mechanisms underlying sensitivity to
organophosphorous anticholinesterase compounds. Progress in Neurobiology., 28: 97-
128.
Venkata Rathnamma V and Nagaraju B (2013). Acute toxicity of Chhlorantraniliprole to
Fresh water fish Ctenopharingodon idella (Valenciennes, 1844). Innovare Journal of Life
Science. Issue 2, 2013, 17-20.

11