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Study on the Effect of Carbofuran in Male Reproductive Organ of Swiss Albino Mice

P. Das
Department of Genetic Engineering and Biotechnology, Rajshahi University, Rajshahi, Bangladesh

M. H. Morshed
Department of Chemistry, Khulna University of Engineering & Technology, Khulna-9203, Bangladesh

A. K. Roy*
Department of Genetic Engineering and Biotechnology, Rajshahi University, Rajshahi, Bangladesh

Abstract/ Executive Summary:

The carbamate pesticide carbofuran which is widely used in fruit, vegetable and crop fields in the world as well as in Bangladesh. The aim of this research work is to assess the toxicant consequences of carbofuran in male reproductive organ through histopathological observation of testis of Swiss albino mice. The experimental mice were grouped into T1, T2, T3 and T4 four groups and the different doses such as 0.5 ppm for T1 group, 1.0 ppm for T2, 1.5 ppm T3 and 2.0 ppm for last group T4 carbofuran were applied to each mice of the respective group. Each of the mice of every group was subsequently sacrificed after 15 days, 25 days and 35 days. The histological slides of treated group of mice were compared with that of control mice and significant changes or abnormalities were observed in the treated mice like cytoplasmic vacuoles in the lethargic epithelium, declension of the cells in the testicular tissue with several cytoplasmic vacuoles, lumen with the loss of spermatid cells. Irregular seminiferous tubules separated from each other with complete degeneration, abnormal seminiferous tubules separated from each other with disintegration and degeneration of spermatogenic cells forming cytoplasmic vacuoles and lumen with loss of spermatid cells, widely separated irregular seminiferous tubules with complete degeneration of spermatogenic cells forming larger lumen with no germ cells. More visible abnormalities were found with the increase of dose and with long term exposure. The result of the study indicates a direct effect on male reproductive organ and may reduce fertility.

Keywords: Pesticide; Reproductive; Histopathology; Fertility; Male Sterility

Location: Department of Genetic Engineering and Biotechnology, Rajshahi University, Rajshahi, Bangladesh

Start Date:

End Date:

Program Area: Pest Management

Commodity: Pesticide

Objectives:

Carbamate insecticide self-poisoning to vary markedly according to the carbamate ingested although the case fatality varied according to the concentration and formulation of the insecticide (Lamba et al., 2016). The industrialization of the agricultural sector has increased the chemical burden on natural ecosystems. Pesticides are agrochemicals used in agricultural lands, public health programs and urban green areas in order to protect plants and humans from various diseases (Stamati et al., 2016). The purpose of the present work was to find out what this could lead to or how it may affect our health or the environment and to develop an animal model for evaluation of the developmental and reproductive effects of early pesticide exposures relevant to human exposures.

Methodology:

The analytical grade carbofuran was collected from alpha agro limited Bangladesh (Purity: 99.99%) and utilized for this experiment. The model animal of the experiments Swiss albino mice (male) collected from the animal rearing house of ICDDRB, Dhaka, Bangladesh at the age of 3 weeks with an average 95gm body weight. Steel cages (47×37×23 cm) were used for rearing and conducting the whole experiment. Suggested especial type of food from ICDDRB was supplied to mice with the standard dose. Freshwater was provided with food wherever necessary. As histopathological tools, microtome machine, paraffin, beaker, blade, spirit lamp, alcohol (100, 95, 90, 70, and 50%), hematoxylin, eosin, Bouin’s fluid, Canada balsam, Mayer's albumin were utilized. The mice (16 nos) were grouped into 5 such as control (4 mice), Treatment T1 (3mice, dose 0.5 ppm), Treatment T2 (3 mice, dose 1.0 ppm), Treatment 3 (3 mice, dose 1.5 ppm) and Treatment 4 (3 mice, dose 2.0 ppm). They were kept in separate cages. Mice of the control group were given only suggested standard food while treatment groups were given carbofuran supplements at a different dose with standard food (Aziz et al., 2007; Pant et al., 1995). Each of the mice of every group was subsequently sacrificed after 15 days, 25 days and 35days. These mice were first anesthetized by diethyl ether and then sacrificed to collect the organs. The collected organs were immediately placed into saline water for washing the debris where they were sectioned into small pieces.

Then these tissues were preserved into Bouin’s fluid for 18 hours. After that period these tissues were washed in the running water to eliminate the trace amount of Bouin’s fluid. Finally, these tissues were preserved into 70% alcohol. The preserved tissues were first taken from the preserving jar. Dehydration is achieved in a series of gradually increasing concentrations of alcohol to reduce some of the shrinkage occurring in the tissue. Dealcoholization took place by xylene. Then the tissues were taken for infiltration which facilitates the penetration of wax to the available spaces inside the tissue. The tissues were then taken into a paraffin bath for roasting. As soon as the tissue is thoroughly filtered with paraffin, it is ready for embedding or block making. After block making, trimming of the block was done. The cutting of the ribbon was done by placing it in a microtome machine. The ribbon was cut at 5 microns by a microtome machine. Then these ribbons were taken into Mayer’s albumin solution water. After this step, the ribbon is placed onto the dry slide. These slides were left for at least 24hours to be dried. This slide was then stained with hematoxylin and then washed with running water. After this process, staining with eosin took place (Gurr, 1962). Motic advanced system biological microscope was utilized to identify the disorder and abnormalities in the cellular structure of replication organ tissue of mice with the help of Motic image J.01 software.

Source of Information: Journal of Engineering Science 10(2), 2019, 85-90

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Remarkable changes were observed in seminiferous tubules of the treated mice organs with tested systemic pesticides, especially with the highest dose T4 in the present study. Degenerations of spermatogenic cells or Sertoli cell fibrosis result in the loss of sperms. The effect also includes necrosis and disintegration of spermatogenic cells, formation of cytoplasmic vacuoles and lumen having no spermatids in the seminiferous tubules. This condition leads to decreased number of sperm counts and abnormal sperms. In summary, carbofuran has adversely affected the reproductive organ of the test animal. The present results indicate that exposure to carbofuran has direct effects on mice testis and the decrease of spermatids may reduce fertility. Carbofuran residue remains inside the food which is being ingested by humans. The laboratory animal utilized in this research protocol showed the strong lethal effect of the used carbamate pesticide carbofuran on the male gamete generating organ. The effect results in decreased reproductively by having a direct effect on germ cell production.

Knowledge Management: Journal