Acute toxicity of percutaneously absorbed malathion, an

organophosphate on Bufo sp. larvae

Joemark Narsico
Natural Sciences and Mathematics Division
University of the Philippines Visayas Cebu College
joemark_narsico@yahoo.com.ph

Abstract

Malathion is commonly used as an insecticide. It is effective in the

inhibition of the enzyme acetylcholinesterase (Navarro et al 2004). Acute
toxicity of malathion was tested using Bufo sp. larvae. One, five, ten fifteen
and twenty ppm concentrations of malathion were prepared through serial
dilution. Observations where recorded for eleven days. The curving of the
tail, head bending, cannibalism and death were exhibited by the Bufo sp.
larvae exposed to malathion concentrations.

Introduction

Malathion diethyl [(dimethylthoxyphosphinothioyl)thio] butanedioate is an

organophosphate, which is widely used as an insecticide because of relatively low
human toxicity. It is present in natural water sources that have been exposed to urban
and agricultural runoff. It is the only organophosphate insecticide that may be applied by
planes to control mosquito populations, and so it also enters water from the air. It is also
widely used in residential landscaping, pubic recreational areasand public health pest
control systems (Relyea et al 2004). However malathion when absorbed or ingested into
the human body readily results to the metabolism of melaoxon which is substantially
toxic. Histopathological study on albino rats conducted by Sesal and colleagues (2008)
showed that erythrocyte cells have defects, degenerative cell size and surface shape in
rats administered with malathion. It was observed that these histopathological defects
were related to the dose increments of malathion on rats.

Amphibian embryos and tadpoles are sensitive at a great number of environmental

pollutants dispersed in the aquatic environment such as pesticides, heavy metals and
polychlorinated biphenyls. Thus, they have been used as a testing system of
environmental contaminants. Amphibian decline have been occurring for decades and
some of these declines have been related to environmental contaminants. Webb and
Crain showed that Xenopus laevis tadpoles exposed to malathion displayed bent tails,
unusual swimming behavior and a higher mortality rate compared to their control group.

Objective

To determine the effects of malathion to Bufo sp. larvae

Materials and Methods

The Bufo sp. larvae were collected from the pond of D’ Family Park, Talamban, Cebu
City. Samples were collected only once for all the treatments. Tadpoles collected were
similar in size and developmental stage. The collected tadpoles were then acclimatized
to distilled water and tap water.

Concentrations of malathion were prepared by serial dilution of technical grade

malathion (570 g/L malathion, 80 g/L emulsifier and 350 g/L xyelene). The prepared
concentrations were 1, 5, 10, 15 and 20 ppm. Distilled water was used to dilute the
malathion. The volume used was 300 mL and it these set-ups were placed In different 5
L jars.

The tadpoles were then placed in the jar. They were not fed. This experiment lasted until
mortality of the control group was observed, which was after 11 days.

The tadpoles were observed for the occurrence of mortality, unusual swimming behavior
and physical malformations every twenty four hours. Dead animals were removed from
the set-up.

Results and Discussion

Tail bending was exhibited in all specimens from day one to the eleventh day. However
there was a high incidence of tail bending from the 15 and 20 ppm. Specimens exhibited
tail bending even on day one when exposed to these higher concentrations. It was
recorded that all set-ups contained a larvae that exhibited tail bending. We also
observed head bending of the larva exposed to malathion concentrations.

A B

Figure 1. Head bending and tail bending of Bufo sp. tadpoles (10x) (A) Head
bending (B) tail bending

A variety of biochemical parameters should be considered as we study the effects of

malathion to the development of the Bufo sp. Malathion is known to inhibit the activity of
acetyl, butylcholinesterase and aliesterase (Rosenbaum et al 1988). These enzymes are
important during cell division. This means that cellular protein synthesis and the
redistribution of these proteins is prevented, thus impeding the normal development of
the larvae. The observations presented were also similar to the observations gathered
by Fordham et al (2000). Fordham presented that the development of malathion
exposed tadpoles were significantly delayed compared to the control group. Malathion
may decrease thyroid function in tadpoles.

Figure 2. Tail curvature. Twenty and fifteen ppm exhibited the highest degree of
tail curvature in the first half of the experiment, however the high malathion
concentration had the highest case of mortality. On the second half of the
experiment no observations recorded since the entire specimen exposed to
higher concentrations died.

Abnormal swimming behavior was also observed among larvae exposed to malathion
concentrations. We also observed that the incidence of abnormal behavior was greatest
at 15 and 20 ppm. Abnormal swimming behavior includes hyperactive symptoms, loss of
balance and motionlessness. Sayim (2007) showed that these are signs of toxicity. The
tadpoles exposed to malathion concentration did not exhibit the maintenance to
equilibrium posture when swimming. Cannibalism was also observed in the experimental
groups although this may be mainly attributed to the starvation of the specimen.

Similar behaviors were also exhibited by the larvae exposed to carbaryl, carbofuran and
dimethoate in amphibian development, which were reported in other studies (Sayim
2007). Sayim further added that these are expected results because malathion and the
other toxins mentioned were neurotoxin which induce abnormal behavioral changes.
When the mentioned abnormal behavior is being exhibited by the tadpole this increases
the risk of tadpole for predation. These symptoms may lead to attraction of predators
and may eventually contribute to the declining population of the amphibians.

Contaminants such as the malathion eventually lead to the death in the Bufo sp. larvae
exposed to malathion concentrations. At day six all of the tadpoles exposed to 15 and 20
ppm died. During the ninth day of experimentation all of the tadpoles exposed to the
malathion concentrations died.
Figure 3. Number of survivors in each set-up from day 6 to day 12.The 20 ppm and
the 15 ppm set-ups has the highest mortality rate on the initial duration of
the experiment.

Prior to the final experimentation, we performed a test run. During the test run we
observed that the mechanical stress and chemical stress due to the change in
environment could have been the cause why mortality was very high during the test run.
But based on the abnormalities and malformations of the embryo we can deduce that
the malathion could have been the cause for the death and abnormal characteristics
exhibited by the tadpole exposed to malathion concentrations.

The amphibian is easily affected to the changes of its environment because it uses much
of its skin for its respiratory function. Polluntants and chemical contaminants could easily
induce stress, abnormal behavior and even death to the amphibian. Amphibians such as
the Bufo sp. are very prone to developing mutations which causes malformed tails and
heads and unusual swimming pattern (Sayim 2007).

Chemical contaminants change the quantity and quality of their food and habitat. From
the observation presented we can deduce that chemical exposure particularly the
malathion could be considered as a contributing factor in the decline of amphibian
populations around the world.

Conclusion

Based on the results obtained, we found that the exposure of Bufo sp. larvae to
malathion could induce physical malformation such as a bent head and tail, unusual
behavior and death of the tadpole exposed to malathion concentrations. However the
concentrations of malathion that resulted to the effects mentioned were generally higher
compared to the reports published on malathion concentration in wetlands.

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