J. Env. Bio-Sci., 2015: Vol.

29 (2):397-399
(397) ISSN 0973-6913 (Print), ISSN 0976-3384 (On Line)

REPRODUCTIVE HISTOLOGICAL ALTERATIONS INDUCED DUE TO MALATHION

EXPOSURE IN FEMALE ALBINO MICE
Sudhir Kumar Kataria* and Shefali
Department of Zoology, Maharshi Dayanand University, Rohtak, Haryana, 124001, India.
[Corresponding author E-mail*: kataria551@gmail.com]

Received: 16-09-2015 Accepted: 20-10-2015

The state of Haryana is an agrarian state which is adjacent to National Capital Territory, New Delhi and is also
known as rice bowl of the country to meet food requirement. Further to enhance food production pesticides are
inevitably used and may results in exposure to non target organisms including human beings. Malathion is an
organophosphorus insecticide widely used in public health, residential, and agricultural practices. In present study,
we investigated the histological alterations caused by orally administered dose of Malathion (761.5mg/kg b.w.
which is 50% of LD50), Cyclophosphamide (40mg/kg of b.w.) and distilled water. Malathion and Cyclophosphamide
exposure results in loss of primordial and primary follicles degenerated ovarian tissue, albuminous fluid in graafian
follicle and change in corpus luteum as compared to negative control which reveals the toxic effects of drug. So it
should be used with prescribed precautions in the interest of healthy sustenance of generations of non target
organisms.

Malathion is an organophosphorus insecticide used in public

health, residential, and agricultural since early as 1950. Over
100 food crops can be treated with Malathion and about half
of total applications in the United States (U.S.) are on alfalfa,
cotton, rice, sorghum, and wheat. In 2004, the Department of
Pesticide Regulation (DPR) reported that 492,307 pounds of
Malathion were used in California1. Malathion has a broad
range of use with target pests belonging to orders like mainly
dipterans, lepidoptera, hemiptera and coleoptera. Malathion
is a slightly toxic compound in EPA toxicity class III carrying
the signal word “CAUTION”.
The histological alterations in different organisms have been
reported in different non target animals other than mice
exposed to different organochlorine and organophosphate
pesticides has resulted in damaged tissues in terms of
anatomical changes2-19. In this study, we investigated the
reproductive histological alterations in gonads (ovary) caused
by Malathion in orally administered female mice as ovary is
an organ sensitive to external factors and has direct bearing
on the next generation.
MATERIAL AND METHODS
Swiss albino female mice (Mus musculus) were obtained from
disease free small animal house of the Lala Lajpat Rai
University of Veterinary Science, Hisar, Haryana, India. Ethical
clearance was taken for the use of mice as experimental
animal from the Institutional Animal Ethics Committee,
Maharshi Dayanand University, Rohtak, Haryana, India. Mice
were acclimatized to the laboratory conditions prior to the
beginning of the experiments.
At the time of dosing each mouse was between 20 - 25 g b.w.
and 8 to 12 weeks old and were divided into three main groups,
group I mice serve as negative control received only the
standard diet and distilled water, group II (positive control)
mice injected cyclophosphamide (40 mg/kg b.w.) in intra-
peritoneal. Group III (treatment group) mice received Malathion
(761.5 mg/kg b.w.). All mice were administered constant
volume of specified doses to study histological alteration in
ovary of mice. All mice were examined immediately after each
dose, approximately 1h for sign symptoms of toxicity and
sacrificed at 24h for histological changes. The female gonad
was removed from the mice and transferred to fresh 10%
neutral buffered formalin and left in it for 24 hrs. The tissue
was then removed carefully and kept under tap water for some
times. Dehydration was done in the ascending grades of
alcohol (50%, 70%, 90% and absolute alcohol) for 30 minutes
in each grade and transferred to xylene. The tissue was then
embedded in paraffinated wax, with the help of a rotary
microtome and thin sections of 5 -6 microns were cut,
spreaded, allowed to stand for a day and deparaffinized in

NAAS Rating (2016)-4.20

Print to PDF without this message by purchasing novaPDF (http://www.novapdf.com/)

 REPRODUCTIVE HISTOLOGICAL ALTERATIONS INDUCED
xylene for 10 to 15 minutes and then hydrated by passing
through a descending alcoholic series and then in distilled
water and stained with Haris Haematoxylin solution and
counter–stained with eosin stain, mounted and observed under
microscope20.
RESULTS AND DISCUSSION
In present study, histological alterations caused by orally
administered Malathion in female mice ovary which is a
sensitive organ to external factors, which might induce
histopathological change as well as functional deficit. The
ovary of negative control mouse showed developing follicles
(primordial, primary and secondary follicles) corpus luteum
and graafian follicles were observed in the cortex of ovum.
Primordial follicles are composed of an oocyte surrounded
by a small number of granulosa cells(Fig.-1) . Our results
revealed that the Malathion treated mice showed a significant
loss of primordial and primary follicles and a significant
reduction of follicles(Fig.-2). The development and maturation
of ovarian follicles involved several stages, including primordial,
primary, secondary, and antral. With the loss of primordial
and primary follicles, all growing follicles (secondary and antral)
were eventually depleted due to the lack of the precursor follicle
populations for recruitment, with impairment to granulosa cells
Figure-1. : Section through the ovary of negative control
mice showing normal structure of graafian
follicles (GF), corpus luteum(CL) & germinal
epithelium.
by the malathion and resulted in anatomical changes in
ovarian follicles. On the other hand, damage induced by
cyclophosphamide were severe with degeneration in ovarian
tissue, albuminous fluid in graafian follicle and change in
corpus luteum was observed in CP treated mice (Fig.-3).
Print to PDF without this message by purchasing novaPDF (http://www.novapdf.com/)
(398)
Figure-2. : Section through the ovary of mice exposed
to 761.5mg/Kg b.w. malathion showing small
damage in epithelial lining, enlarged corpus
luteum (CL), small change in number of atretic
follicle.
Histopathology is the gold standard when defining toxicological
effects. Using biomarkers linked to distinct, defined cell types
and tissues may provide a direct link to histopathology without
its drawbacks and it also provides increased sensitivity and
specificity21. Histopathological biomarkers are closely related
to stress since many pollutants either toxic or non toxic have
to undergo metabolic activation in order to be able to culminate
cellular change in the affected organism. The mechanism of
action of several xenobiotics could initiate the formation of a
specific enzyme activity that causes changes in metabolism,
further leading to cellular intoxication and finally death22-23.
Reports are not available on ovarian damage due to exposure
to malathion toxicity in female mice, however, in mammals
and other groups it has been reported that administration of
malathion (33mg/kg/day) in rats cause atresia accompanied
by a decreased number of normal antral and growing follicles
in ovary, area of corpus luteum was enlarged due to dose
increments24. The study revealed that Malathion affects
numbers of primordial, primary and prenatal follicles. In another
study, exposure dependent alterations in ovary for both acute
and chronic exposures were reported in fish due to sub lethal
doses of Malathion where reduction in size of mature oocytes,
disruption and vacuolation in cytoplasm after acute exposure,
whereas chronic exposure resulted in complete loss of normal
configuration of ovary, necrosis, elongated ovarian follicles,
and fragmented ova with abnormal shape17. The notable
 (399) KATARIA AND SHEFALI

3. Konar, S.K. (1970). J. Inland. Fish. Soc. India., 2: 5 1.

4. Natarajan, G.M. (1981). Pest. Biochem. Physiol., 21, 194.
5. Girija, M. (1987). In: Ph/D. thesis submitted to S.V. Univ., Tirupathi,
A.P., India.
6. Anita, S. T. (1994). In: Ph.D. thesis submitted to Nagarjuna
Univ., A.P., India.
7. Dudhat, I.N. (1996). J Aqu. Bio.,11(1&2)67.
8. Ramana Kumari, G. V. (1999). In: M.Phil. dissertation submitted
to Nagarjuna Univ., Guntur, India.
9. Karuppasamy (2000). Bull. Pure and Appl. Sci, 19A (2):109.
10. Ramachandra, M. M. (2000). Polln. Res., 19(1): 73.
11. Tilak, K. S. and Yacobu, K. (2002). J.Ecotoxicol. Environ.
Monit., 12(1): 09.
Figure-3. : Section through the ovary of mice exposed 12. Sabina, T.l., Przebirowska, D.O., Latuszynska, J., Rodak, M.T.
to 40mg/Kg b.w. cyclophosphamide showing and Maj, A.H. (2003). Ann. Agric. Environ. Med., 10:101.
enlarged corpus luteum (CL), degeneration in 13. Tilak, K. S., Veeraiah, K. and Koteswara Rao, D. (2005). J.
ovarian tissue, albuminous fluid in graafian Environ. Biol., 26 (2 suppl): 341.
14. Tilak, K. S., Veeraiah, K. and Thathaji, P. B. (2007). J. Ecotoxicol.
follicle.
Environ. Monit.,17(2): 129.
microscopic changes in oocytes at different stages of 15. Al-Attar, A.M .(2010). J. Biomed. and Biotech.,1.
development and in the nucleus of the immature oocyte of 16. Ksheerasagar R. L., Hiremath M.B. and Kaliwal B. B. (2011). J.
the catfish Heteropneustes fossilis25. Malathion affects number Sci and Tech., 1(5): 43.
of primordial, primary and prenatal follicles, but it increases 17. Magar, R.S. and Bias, U.E. (2013). Int Res J of Env S., 2(3), 59.
atretic follicles and corpus luteum. It is reported that the 18. Slimen, S., Saloua el, F. and Najoua, G. (2014). Iran J. Basic
treatment of female mice with CP (120mg/kg/bw) resulted in Med Sci., 17(7):522.
severe ovarian damage26. 19. Omran, O.M., Omer, O.H. (2015). Pathol. Res. Pract., (6):462.
20. Luna, LG. (1968). In: Manual of histologic staining methods of the
In conclusion, it is clear that the pesticide exposure causes armed forces institute of pathology. 3rd Ed. New York: McGraw-Hill;
hazardous effects at various levels to non-target organisms, 21. Kilty, C.G., Keenan, J. and Shaw, M. (2007). Expert Opin Drug
including humans. The state of Haryana being agrarian state Saf., (2):207.
needs to take specific steps to educate farmers about its ill 22. Velkova-Jordanoska, L. (2002). In: MSc thesis submitted to

effects so that extent of toxicity be minimized to its Univ. St.Kiril and Methodius, Skopje, R. Macedonia (Mc).
23. Roganovic-Zafirova, D., Jordanova, M., Panov, S. and Velkova-
inhabitants.
J ordanos ka, l. (2003). In: Hepatic capillariasis in the
ACKNOWLEDGEMENTS Mediterranean barbell (Barbus meridionalis petenyi Heck.)

Authors are grateful to the Department of Zoology, M.D. from Lake Ohrid. Folia Veterinaria, 47(1):35.
24. Koc, N.D., Kayhan, F.E., Sesa, C. and Muslu, M.N. (2009). Pak.
University, Rohtak for providing necessary facilities for this
J. Bio. Sci., 12 (6):498.
research work.
25. Dutta, H.M., Nath, A., Adhikari, S., Roy, P.K., Singh, N.K. and
Datta Munshi, J.S. (1994). Hydrobiologia., 294 (3): 215.
REFERENCES 26. Jiang, Y., Zhao, J., Qi, H.J., Li, X.L., Zhang, S.R., Song, D.W., Yu,
C.Y, and Gao, J.G. (2013). J. Zhejiang. Univ. Sci. B., (4):318.
1. California Department of Pesticide Regulation’s Pesticide Use
Database. (2004).
2. Lowe, J.I. (1966). In: Proc. 19th Annual Conf. S.E. Assoc. Game
and Fish Comrn.

Print to PDF without this message by purchasing novaPDF (http://www.novapdf.com/)