Egypt J. Med. Lab. Sci., Sept. 2007; 16(2): 99 -1131110-5593

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THE EFFECT OF AN ORGANOPHOSPHOROUS INSECTICIDE ON THE HEPATIC, RENAL AND PULMONARY TISSUES OF MICE FETUSES

Fawzy I. Amer, Iman Zakaria, Hamza A. El-Shabaka and Ihab Ashour

Department of Zoology, Faculty of Science, Ain Shams University

ABSTRACT

Introduction: The present study deals with the effects of the organophosphorous insecticide curacron on the hepatic, renal and pulmonary tissues of mice fetuses. Materials and Methods: Fourty pregnant females were divided into four groups, 10 individuals each. The 1st group served as control group. The individuals of the 2nd and 3rd groups were given oral dose of curacron 14.4 and 28.8 mg/kg, respectively on day 7 of gestation. The 4th group was given 14.4 mg/kg for 8 successive days from day 7 to day 14 of gestation. At the 18th day of pregnancy, females from both the control and treated groups were sacrifi ced and samples of hepatic, renal and pulmonary tissues were taken from the fetuses.Results: The liver of the 18-days-old fetuses of all experimental groups showed different phases of venous congestion, disorganization of the hepatic lobules, fatty degeneration and necrosis. The kidney of curacron-treated groups showed venous congestion, cloudy swelling, and necrosis of the epithelial cells lining the kidney tubules. Moreover, the lungs of the different treated groups showed dilation and congestion of the blood vessels, thickening of the interalveolar septa and scattered cell debris in the lumens of the alveoli.Conclusion: Consequently, cumulative doses of curacon obviously affect hepatic, renal and pulmonary fetal tissues and extreme caution should be considered by pregnant women to avoid its hazardous effects on their fetuses.Key Words: Organophosphorous, Insecticides, Curacron, Mice fetus liver, Kidney and Lung.

Corresponding Author: Dr. Fawzy I. Amer, Department of Zoology, Faculty of Science, Ain Shams University, Abassia, Cairo, Egypt.

INTRODUCTION

Pesticides have contributed to dramatic in-creases in crop yields, and in the quantity and variety of the diet. They have helped to limit the spread of certain diseases, but can cause injury to human health as well as to the envi-ronment (Mansour, 2004). The major predic-tors of health risk from pesticide exposure are quantity and toxicity of pesticides reaching end-users, fi eld workers and persons with ca-sual and indirect exposures to fi eld and food residues, drift and contaminated groundwater (Richter and Safi , 1997). On the other hand exposure to very low doses of common pesti-cides, at levels currently assumed to be safe and within dose ranges that have been mea-sured in people, caused signifi cant injury to mice embryos tested in vitro during early stag-es of development (Greenlee, et al. 2004).

Coulombe et al. (1986) studied the pulmo-nary toxicity of the organophosphorous insec-ticide fenitrothion following a single exposure of rats to the fi eld formulation, at the site of an aerial spraying. Rat lungs examined under light and electron microscopy showed a few signs of toxic lung injury. The alveolar toxic reaction was limited to small and discrete foci and was entirely reversible within a period of 2 months. Enan et al. (1986) studied the effect of 1/10 LD50 of profenofos on the liver and kidney of rats and reported that, congested blood vessels, nodular cellular infi ltration and dense fi brosis around the blood vessels ap-peared 1,2 and 3 weeks, respectively following exposure. The kidney tissue showed hyalini-zation affecting some glomeruli and the be-ginning of degeneration in the related tubules

Original Article

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followed by dilation of the proximal and distal tubules. Moreover, El Nahas et al. (1988) in-vestigated the cytogenetic effects of curacron on mice somatic cells and reported that cu-racron signifi cantly increased the frequency of chromosome aberrations and also inhib-ited the mitotic activity. Meanwhile, El Sebae et al. (1988) declared that profenofos showed maximum effect on protein synthesis in rab-bit liver and muscle tissues at a dose of 0.2 microgram/ml.

Moreover, El-Swak (1990) found that histopathological examination revealed widespread congestion and degenera-tive changes in the organs and tissues of hens after a single oral dose of profenofos (25mg/kg). Saeed et al. (1995) stated that profenofos caused different symptoms of tox-icity and revealed some biochemical changes especially in the enzymes activity of the liver following two sublethal doses of profeno-fos to pregnant female Swiss mice during organogenesis. On the same line, Ibrahim and Sayed (1998) reported that there was a signifi cant decrease in the mean number of live fetuses and the body weight of fetuses of all treated mice following the same doses of profenofos used by the former authors.

Abdel Hadi and Abedin (1997) studied the histopathological changes in the liver and kidney tissues of mothers and newborn rats, Rattus rattus, after treatment with Dimethoate or Nuvacron insecticides. Results showed marked degenerative changes including lymphocytic infi ltrations and nuclear pleomorphism. Moreover, high doses of the insecticides induced tissue vacuolization, haemorrhage and hyperplasia of Kupffer cells in the liver. In addition, swelling of Bowman's capsules and tubular degeneration in the kidney were found. Finally, they suggested that the insecticides seem to affect newborn through the transmission of their chemicals through mother's placenta. Fahmy and Abdalla (1998) indicated that the two pest control agents buprofezin and petroleum oil are relatively much safer compounds than profenofos. Chromosomal aberration induced by profenofos in somatic and germ cells of male mice were highly signifi cant than by the other two agents.

Recently, Mahadevaswami and Kaliwal (2003) investigated the effect of dimethoate, an organophosphorous insecticide and pro-gesterone on implantation in virgin pregnant albino mice. They stated that treatment with dimethoate for 7 days (from days 1 to 7) or with dimethoate for 7 days followed by progester-one for 8 days (up to day 15) totally abolished implantation, with a 100% pre-implantation loss. Munidasa et al. (2004) studied the out-comes and predictors of mortality in patients with acute organophosphates (OP) poisoning requiring intensive therapy over a period of 40 months. They reported that mortality following (OP) poisoning remains high, of the 71 pa-tients studied 36 had died, despite adequate respiratory support and intensive care. Farag et al. (2007) studied the effect of organophos-phate insecticide dimethoate at three dosage levels on male reproduction in mice. Brain and skeletal muscle acetylcholinesterase ac-tivities were inhibited both in the middle and high dose groups. Dimethoate was associat-ed with a decreased number of implantations and live fetuses and an increased number of dead and early resorptions at high dose-treated group. Sochaski et al. (2007) inves-tigated the blood fenitrothion concentrations in the rat dam and foetus and tissue esterase activity following in-utero exposure to the or-ganophosphate insecticide fenitrothion. They reported that fenitrothion concentrations in maternal and foetal blood were virtually identical. The authors added that esterase inhibition occurred at a fenitrothion dose of 5mg/Kg, that has not previously associated with reproductive toxicity, suggesting that such inhibition should be considered as the critical effect in risk assessment for this pesticide.

Thus, a need has sprung for an accu-rate and critical examination of the possible teratogenic impacts of many insecticides related to different chemical groups. The purpose of the present study is to evalu-ate the teratogenic effects of curacron on the hepatic, renal and pulmonary tissues of 18-days-old CD1 mice fetuses.

MATERIAL AND METHODS

Adult males and females of CD-1 Swiss al-bino mice with an average body weight of 25-30g were obtained from the breeding unit of

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poorly developed. The hepatic lobules are composed of irregular branched and intercon-nected hepatic strands of 1-3 cells thick radiat-ing from the central vein to the lobule periphery (Figures 1 and 2).

The hepatocytes are more or less polygo-nal in shape with distinct boundaries posses-ing spherical nuclei (Figure 2). The blood si-nusoids are irregularly dilated vessels, lined with a discontinuous layer of endothelial cells intervened by large phagocytic Kupffer's cells (Figure 2). As the liver in the late embry-onic stages acts as a hemopoietic organ, a considerable number of different stages of erythroblasts and few megakaryocytes are found in between the hepatic cells (Figure 2).

Theodor Bilharz Research Institute, Imbaba-Cairo, Egypt. Three adult females in pro-es-trous stage were housed together with an adult male in the same cage for overnight. Vaginal smears were then examined in the next morn-ing and the presence of spermatozoa was considered as an indication of pregnancy and the date was considered as 0 day of gestation.

The insecticide used in the present in-vestigation was the organophosphorous in-secticide, curacron (Profenofos), obtained from Ciba-Geigy agrochemicals division, Dokki, Cairo.Desired concentrations were prepared using distilled water as a diluent.

Fourty pregnant females were used in the present study. They were divided into four groups of 10 individuals each. The fi rst group (GI) was used as control. The pregnant fe-males of the second group (GII) were given, each, a single oral dose of 14.4 mg/kg body weight of curacron at day 7 of gestation. The pregnant mice of the third group (GIII) were given, each, a single oral dose of 28.8 mg/kg body weight of curacron at day 7 of gestation, while the pregnant females of the forth group (GIV) were given, each, a daily oral dose of 14.4 mg/kg body weight of curacron for 8 suc-cessive days, from day 7 to day 14 of gestation.

At the 18th day of pregnancy females from both the control and the treated groups were sacrifi ced. The uteri were removed and fe-tuses were fi xed in aqueous Bouin’s solution for at least 24 hours and then preserved in 70% ethyl alcohol. The fetuses were dehy-drated in ascending grades of ethyl alco-hol, cleared in terpineol and embedded in paraffi n wax. Serial transverse and sagit-tal sections 5 microns thick of fetuses were cut, mounted and stained with haematoxylin and eosin for general histological studies.

RESULTS

THE LIVER

Control groupExamination of the liver of 18-days-old con-

trol fetus (group I) revealed that it is formed of homogenous mass of parenchyma cells arranged in hepatic lobules separated by in-terlobular connective tissue septa which are

Figs. 1 and 2: Photomicrographs of sections of the liver of 18-days-old control fetus (Group I).

Fig. 1: Showing the hepatic lobules that can be only distinguished by their central veins, hepatocytes and blood sinusoids. X 100

Fig. 2: Showing the central vein with its intact endothelial lining (arrow head), the blood sinusoid and Kupffer's cells (arrows) protruding into the cavity of the blood sinusoid. X 400

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THE EFFECT OF AN ORGANOPHOSPHOROUS INSECTICIDE ...

Treated GroupsExamination of the liver of 18-days-

old fetus maternally treated with 14.4 mg/kg body weight of curacron at day 7 of gestation, G (II) revealed no remarkable changes as compared with that of the con-trol (Figures 3 and 4). Erythroblasts have normal appearance, however, some are ag-gregated inside the blood sinusoids (Figures 3 and 4). In addition, a marked increase in the number of megakaryocytes with their characteristic, lobulated, voluminous nuclei are more frequently encountered (Figure 4).

Examination of the liver of 18-days- old fetus maternally treated with 28.8 mg/kg body weight of curacron at day 7 of gestation, G (III) showed that, the majority of the liver cells have no remarkable changes and their vesicular prominent nuclei appear normal. However some hepatocytes showed histopathological alterations including both fatty degeneration and necrotic hepatocytes (Figure 5). Degeneration was reported by

the presence of numerous vacuoles in the cytoplasm of these cells. Moreover, the nu-clei of these cells are either pyknotized or karyolysed (Figure 5). Sinusoids are dilated and some of them are congested. These sinusoids contain stagnant blood with in-tact and hemolysed blood cells (Figure 5).

Examination of the liver of 18-days-old fetuses maternally treated with repeated oral doses (14.4 mg/kg) of curacron for 8 successive days starting from day 7 to day 14 of gestation, G (IV) revealed severe histopathological alterations (Figures 6 and 7). The most striking effects of the repeated doses of curacron were the congestion of blood vessels and sinusoids

Figs. 3 and 4 : Photomicrographs of sections of the liver of 18-days-old fetus maternally treated with 14.4 mg/kg of curacron (Group II).

Fig. 3 : Showing normal architecture of the liver tissue. X 400

Fig. 4: Showing the normal appearance of hepatocytes, blood sinusoids, normal erythroblasts and megakaryocytes. X 400

Fig. 5: Photomicrograph of a section of the liver of 18-days-old fetus maternally treated with 28.8 mg/kg of curacron (Group. III), showing congestion of sinusoids (asterisks) and some hepatocytes exhibit vacuolar degeneration. Pyknotised (arrow head), karyolysed (thick arrow) and karyorrhexis (arrow) of some liver cells are found. X 400

as well as the appearance of scattered focal areas of infl ammatory lymphatic and eosinophilic cell infi ltration in the hepatic tissue (Figure 6). Most of the central veins were congested and contained stagnant intact and hemolysed blood cells (Figure 6).

The liver of this group showed marked loss of the lobular architecture and disorganiza-tion of the hepatic strands. Histopathological alterations including both fatty degeneration and necrosis were observed (Figure 6). These degenerated hepatocytes possessed pykno-tized, karyorrhexed or karyolysed nuclei.

Blood sinusoids were markedly dilated with marked aggregation of erythroblasts and in-crease in the number of the megakaryocytes (Figure 7) was encountered. Hence the repeat-

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represents a renal lobe which consists of the uriniferous tubules and stromal tissue. The latter is formed of a network of connective tissue (Figure 8).

The uriniferous tubule is composed of the nephron which is formed of the Malpighian corpuscle, the proximal convoluted tubule, the descending and the ascending limbs of Hen-le's loop and the distal convoluted tubule. Each corpuscle consists mainly of a tuft of blood capillaries, or glomerulus and a Bowman's capsule which is a double walled cup formed of two layers, an outer parietal layer and an inner visceral layer separated by a clear, dis-tinct capsular space (urinary space) (Figure 9).

The proximal convoluted tubules ap-pear rounded, and is lined by a single layer of short columnar cells with indistinct cell boundaries, and spherical nuclei. The free ends of these cells have well-developed brush borders that almost fi ll most of the lumen (Figure 9).The distal convoluted tu-bules are lined with simple cuboidal epithe-lial cells that possess distinct cell boundaries and a granular cytoplasm, and conspicuous spherical centrally located nuclei (Figure 9).

Treated groupsExamination of the cortical region of the

kidney of 18-days-old fetus maternally treated with 14.4 mg/kg body weight of curacron at day 7 of G (II) gestation, revealed various histopathological changes. Most of the renal corpuscles appeared more or less normal. However, some glomeruli have lost their normal circular shape and converted into shrunken, abnormally cellular and relatively vascular structures having few red blood cells and leaving rather wide urinary space (Figures

ed doses of curacron seemed to accelerate the formation of erythroblasts and megakaryo-cytes. Most of the endothelial cells lining the sinusoids were markedly eroded and ruptured. Kupffer's cells became hypertrophied with enlarged and faintly stained nuclei (Figure 7).

THE KIDNEY

Control groupExamination of the kidney of 18-days-

old control fetus G(I) revealed that it is differentiated into an outer cortex and an inner medulla, which is formed of conical pyramids. The cortex shows radial striations, the medullary rays are extending from the base of the pyramids without reaching the renal capsule. Each medullary pyramid with the corresponding part of the cortex

Figs. 6 and 7: Photomicrographs of sections of the liver of 18-days-old fetus maternally treated with 14.4 mg/kg of curacron (Group IV).

Fig. 6 Showing congestion of the central vein, the surrounding tissue is infi ltrated with many infl ammatory cells. The hepatocytes exhibit severe fatty degeneration. X 250

Fig. 7: Showing the increased number of encountered megakaryocytes and erythroblasts. Necrotic areas (asterisk) are scattered all over the tissue. X 400

Fig. 8: Showing the renal capsule (arrow), outer cortical and inner medullary regions and the medullary rays. X 40

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10 and 11). The lining cells of the proximal and distal convoluted tubules exhibited cloudy swelling. They became swollen and projected inwards attaining a conical shape. The lumina of these tubules became relatively small. Some cells showed advanced degenerative changes and their nuclei were pyknotized or karyolysed (Figures 10 and 11).

Examination of the cortical region of the kidney of 18-days-old fetus maternally treated with 28.8 mg/kg body weight of curacron at day 7 of G (III) gestation, revealed distinct pathological lesions in the urineferous tubules. The changes which occurred in the glomeruli were manifested by their hypertrophy probably due to their congestion and the mild proliferation of their constituent cells. In addition some other glomeruli appeared shrunken with wide urinary

spaces (Figure 12). The lining cells of the convoluted tubules exhibited cloudy swelling, hydropic degeneration and cellular necrosis. The outer borders of the cells lining the convoluted tubules were deteriorated, their lumina contained faintly stained casts and the brush borders of the proximal convoluted tubules were destructed (Figure 12).

Examination of the cortical region of the kidney of 18-days-old fetuses maternally treated with repeated oral doses (14.4 mg/kg) of curacron for 8 successive days starting from day 7 to day 14 of gestation G (IV), revealed severe histopathological alterations. The glomeruli exhibited clear features of damage represented by congestion of their capillaries which were packed with red blood corpuscles (Figure 13). Vacuolation of the cytoplasm of the cells lining the convoluted tubules was obvious and the nuclei of some deteriorated cells showed obvious signs of pyknosis (Figures 13 and 14). Some of the lining cells of the convoluted tubules and collecting tubules showed features of necrosis. These cells were detached in the lumina of the tubules and their nuclei were either pyknotized or completely disappeared (Figures 13 and 14). Intertubular extensive hemorrhagic areas were obviously noticed among the tubules (Figures 13 and 14).

The medullary region of the kidney of this group showed, marked degenerative changes in the descending and ascending

Figs. 8 and 9: Photomicrographs of sections of the kidney of 18-days-old control fetus (Group I).

Fig. 9: Showing a part of the cortical region containing, a renal corpuscle and both proximal and distal tubules. X 250

Figs. 10 and 11: Photomicrographs of sections through the cortical region of the kidney of 18-days-old fetus maternally treated with 14.4 mg/kg of curacron (Group II) .

Fig. 10: showing, cloudy swelling of the epithelial lining of the proximal and distal convoluted tubules (arrow heads), shrunking of the glomerular tuft (asterisk) and widening of the urinary space (arrow). X 250

Fig. 11: Showing that the glomeruli contain few red blood corpuscles (asterisk) and the lining cells of both the proximal and distal tubules exhibit different stages of necrosis and pyknotized (arrows) or karyolysed (arrow head) nuclei. X 400

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limbs of Henle's loop as well as the collect-ing tubules. Such changes included cloudy swelling, hydropic degeneration and cel-lular necrosis. The lumina of the collect-ing tubules contained faintly stained casts. The intertubular connective tissue con-tained small hemorrhagic areas (Figure 15).

THE LUNG Control groupExamination of the lungs of 18-days-old

control fetus (G I) revealed that the pulmo-nary tissue consists of the bronchial tubes, respiratory bronchioles and alveolar ducts which lead into many blind alveolar sacs. The pulmonary alveoli are lined with simple squamous epithelium and are surround-ed by a network of pulmonary capillaries (Figure 16). The alveoli are separated from each other by interalveolar septa which consist of three main types of cells, the endothelial cells of the blood capillaries, the squamous alveolar epithelial cells and the connective tissue cells or septal cells (Figure 17). The endothelial cells lining the blood capillaries are elongated with intensely stained elongated nuclei. The squamous al-veolar epithelial cells are characterized by their intensely stained cytoplasm and elon-gated faintly stained nuclei. The connective tissue cells are irregular, oval or cuboidal in shape and possess faintly stained cytoplasm and large oval or rounded nuclei (Figure 17).

Treated groupsExamination of the lung of 18-days-old

fetuses maternally treated with 14.4 mg/kg

Fig. 12: Photomicrograph of a section through the cortical region of the kidney of 18-days-old fetus maternally treated with 28.8 mg/kg of curacron (Group. III) showing, cloudy swelling of the epithelial lining of the proximal and distal convoluted tubules (arrows), casts inside the lumina of most of the tubules (arrow heads) and hypertrophied congested glomerular tufts (asterisk). X 250

Fig. 13: Showing, degeneration of the cytoplasm and deterioration of the nuclei of the lining cells of the proximal and distal convoluted tubules (arrows), intertubular hemorrhagic areas (asterisks), faintly stained casts inside the lumina of most tubules (arrow head). X 250

Fig. 14: Showing, lobulated and congested glomerular tufts (arrow heads), intertubular hemorrhagic area (asterisk), faintly stained casts in the lumina of some tubules (arrow) and vacuolation of the cytoplasm of the cells lining the convoluted tubules (thick arrow). X 250

Figs.13-15: Photomicrographs of sections through the cortical region of the kidney of 18-days-old fetus maternally treated with 14.4 mg/kg of curacron (Group. IV).

Fig. 15: Showing degenerative changes in the descending and ascending limbs of Henle's loop as well as the collecting tubules. Many of these cells show necrosis and pyknotized nuclei (thick arrows). Notice the presence of small intertubular hemorrhagic areas (arrow heads). The lumina of the tubules contain few faintly stained casts (long arrows). X 250

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THE EFFECT OF AN ORGANOPHOSPHOROUS INSECTICIDE ...

body weight of curacron at day 7 of gesta-tion G(II), revealed variable histopathological changes including congestion of pulmonary blood vessels and capillaries (Figures 18 and 19), thickening of the interalveolar septa sep-arating the pulmonary alveoli with infi ltrating lymphocytic or infl ammatory cells (Figures 19 and 20). The cells lining the respiratory bronchioles possessed vacuolated cyto-plasm (Figure 19). In addition, the epithelial lining of the respiratory bronchioles and pul-monary alveoli was partly shed and few cells were detached in their lumina (Figure 20).

Examination of the lung of 18-days-old fe-tus maternally treated with 28.8 mg/kg body weight of curacron at day 7 of gestation G (III), revealed remarkable histopathological changes as compared with the control group. Marked pathological lesions were observed in the pulmonary tissues which included dila-tion and congestion of the pulmonary blood

vessels and capillaries (Figures 21 and 22). The lining epithelium of the terminal bron-chioles and respiratory bronchioles was severely destructed and many cells could be seen detached in their lumina (Figure 21). In addition, the cells lining the respira-tory bronchioles possessed vacuolated cy-toplasm and pyknotic nuclei. The alveolar walls were slightly thickened and the interal-veolar septa contained large number of infi l-trated lymphocytes as well as red blood cells.

Examination of the lung of 18-days-old fe-tuses maternally treated with repeated oral doses (14.4 mg/kg) of curacron for 8 suc-cessive days starting from day 7 to day 14 of gestation G(IV), revealed severe histo-pathological alterations. The blood vessels and capillaries were highly congested and their lumina were fi lled with blood cells (Fig-ures 23 and 24). The mucosal layer of the respiratory bronchiole was thickened, folded

Figs. 16 and 17: Photomicrographs of sections of the lung of 18-days-old control fetus (Group I)

Fig. 16: Showing the respiratory bronchioles lined with cuboidal epithelium (arrow), the alveolar sacs, alveoli, and the interalveolar septa. X 250

Fig. 17: Showing the alveolar wall composed of three types of cells; the squamous alveolar epithelial cells (P1), the connective tissue cells or septal cells (P2) and the endothelial cells of the blood capillaries (P3). X 400

Fig. 18: showing dilated and congested blood vessel with stagnant and hemolysed blood (large asterisk), thickening of the interalveolar septa (small asterisks) and detached cells in the respiratory bronchioles (arrows). X 100

Fig. 19: Showing congested blood vessel (asterisk),the lining cells of the respiratory bronchioles show vacuolation (arrows) and thickening of the interalveolar septa (large asterisk). X 250

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and contained darkly stained cells with py-knotized nuclei. In addition, the lining epithe-lium of some bronchioles and alveolar ducts was destructed and numerous cells were detached in their lumina. The interalveolar walls were abnormally thickened due to the extensive development of the epithelial cells (hyperplasia). These interalveolar walls con-tained large number of lymphocytic infl am-matory cells and red blood cells (Figures 24 and 25). The cells of the interalveolar septa possessed highly vacuolated eosinophilic cy-toplasm and pyknotised nuclei (Figure 25).

Figs. 18-20: Photomicrographs of sections of the lung of 18-days-old fetus maternally treated with 14.4 mg/kg of curacron (Group II).

Fig. 20: Showing that the alveoli contain some hemolysed blood cells (asterisk), the interalveolar walls are thickened with many lymphocytes (arrows) and detached cells inside the alveolar lumina (arrow heads). X 250

Figs. 21 and 22: Photomicrographs of sections of the lung of 18-days-old fetus maternally treated with 28.8 mg/kg of curacron (Group. III).

Fig. 21: Showing detached cells at the inner border of the respiratory tubules (arrows), congested blood vessels (asterisks) and thickened interalveolar septa (arrow heads). X 100

Fig. 22: Showing thickened interalveolar septa with many lymphatic infl ammatory cells (arrow), the alveolar sacs are fi lled with hemolysed blood cells and hemorrhage (asterisks). X 250

Fig. 23: Showing highly congested blood vessels with hemolysed blood cells (asterisks), with many infi ltrating lymphocytes, the walls of the respiratory bronchioles are highly destructed and their lining epithelia are folded. Note hyperplasia of the interalveolar septa with an abnormally increased number of lymphocytes (arrows). X 100

Fig. 24: Showing hyperplasia of the interalveolar septa with an abnormally increased number of lymphocytes (asterisks) and the highly folded inner border of the respiratory bronchioles with some detached cells (arrows). X 250

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LIST OF ABBREVIATIONS

AL : AlveoliAL.D : Alveolar ductAL.S : Alveolar SacAS.L : Ascending limb of Henle's loopBL.S : Blood sinusoidBL.V : Blood VesselBO.CP : Bowman's capsuleCO.T : Collecting tubuleCV : Central veinCX : CortexDS.L : Descending limb of Henle's loopDT.T : Distal convoluted tubuleEB : ErythroblastGL : GlomerulusHC : HepatocyteIAL.SP : Interalviolar SeptumIPR : InterparietalMD : MedullaMD.R : Medullary rayMG : MegakaryocyteN : NucleusPX.T : Proximal convoluted tubuleR.B : Respiratory bronchioleUR.S : Urinary space

DISCUSSION

Organophosphates (OPs) are some of the most us eful and diverse classes of in-secticides in use for almost fi ve decades. Although they continue to be extremely useful in agricultural pest control through-out the world, yet, their extensive use has led to numerous poisonings of non-target

Fig. 25: Showing thickening of the interalveolar septa with large number of lymphocytes (asterisks), vacuolation of the cells of the interalveolar walls (arrows) with many pyknotized nuclei (arrow heads) and many alveolar lumina containing detached cells (thick arrows). X 400

Figs. 23-25: Photomicrographs of sections of the lung of 18-days-old fetus maternally treated with 14.4 mg/kg of curacron (Group IV).

species. OPs ability to poison insects is the result of their anticholinesterase activity (El-Masry et al. 1985 ; Goh et al. 1990 ; Seno et al. 1998 ; Gomes et al. 1999 ; Farag et al. 2003). There is, however, a growing body of literature indicating that organophosphorous compounds may induce oxidative stress lead-ing to generation of free radicals and altera-tions in antioxidants and scavengers of oxy-gen free radicals (OFRs) (Akhgari et al. 2003).

Meanwhile, organophosphates have been shown to cross the placental barrier (Vil-leneuve et al. 1972) and thus could poten-tially affect the developing fetus as inves-tigated by many authors (Abdel Hadi and Abedin, 1997 ; Farag, et al. 2000 ; Bonfanti et al. 2004 ; Sochaski, et al. 2007).

Saad et al. (1992) studied the effect of drift-ed profenofos residues on clover, on the New Zealand white male rabbits, where profenofos residues in dorsal muscles and liver were de-termined after heating. In the present study, the liver tissues of 18-days-old fetuses mater-nally treated with different doses of curacron revealed some histopathological changes. The results were dose-dependant which agrees with Abdel Hadi and Abedin (1997). Single low dose administration (group II) of the present study revealed only slight histo-pathological changes including aggregation of non-nucleated red blood cells inside the blood sinusoids and an increase in the number of the megakaryocytes. This contradicts with the results that have been observed in rats by Chung et al. (2002) who reported that at low-er doses of fl upyrazofos there were no signs of either maternal toxicity or embryotoxicity.

The hepatic tissues of fetuses of Group III and Group IV showed various histopathologi-cal alterations including loss of the lobular ar-chitecture and disorganization of the hepatic strands, fatty degeneration and necrosis of the hepatocytes that possessed many py-knotized, karyolyzed or karyorhexed nuclei. Blood sinusoids were dilated and congested and their endothelial lining was markedly eroded and ruptured. Congestion of the cen-tral veins, invasion of many infl ammatory cells and hypertrophy of kupffer cells were recorded besides the marked increase in the number of the encountered megakaryocytes

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and erythroblosts generally. These results are in agreement with those obtained by Bay-oumi et al. (1979) who reported that diazinon, an organophosphorous insecticide, caused fatty degeneration and pyknosis or karyor-rhexis of the hepatocytes after long term ad-ministration. In addition, Mikhail et al. (1979) observed liver necrosis of mid-zonal type and fatty changes after two repeated intraperito-neal injections of 1/2 the LD50 of dursban to albino rats. In this respect, fatty degenera-tion, necrosis and vacuolation of hepatocytes in fi shes were reported by Desai et al. (1984).

The degenerative changes of the hepatic tissues observed in the present study, includ-ing lymphocytic infi ltrations, nuclear pleomor-phism, vacuolization, hemorrhage and hyper-plasia of Kupffer cells, confi rm the results of El Elaimy et al. (1995) and Abdel Hadi and Abedin (1997). Moreover, Karim (1998) ob-served that liver of rats fetuses maternally treated with fl ufenoxuron showed vacuolation of hepatocytes and dense focal cellular infi l-tration with lymphocytes and polymorph cells.

On the other hand, it is worth mention-ing that the congestion of the central veins and other blood sinusoids may be attributed to heart failure as a result of the adminis-tration of organophosphorous insecticides as reported by Yen et al. (2000) who sug-gested that patients with acute respiratory failure, induced by severe organophospho-rous poisoning, showed symptoms of heart failure. In addition, Haschek and Rousseaux (1991) explained that heart failure produces changes in different organs due to the ex-cessive blood in venous system which in-creases the blood pressure in the veins and capillaries, thus exerting a pressure on the neighboring structures. Meanwhile, arterial blood pressure is reduced and the organs receive a diminished blood supply leading to malnutrition, hypoxia and the accumula-tion of excretory and metabolic products.

In the present study the kidney of 18-days-old fetuses maternally treated with curacron (groups II,III and IV) showed several histopathological changes. The cortical region showed shrinkage of the glomerular tufts and widening of the urinary space. Dramatical changes were noticed in groups III and IV

respectively, including hypertrophied and congested glomerular tufts, degeneration and vacuolation of the cytoplasm of the cells lining the convoluted tubules. Moreover, the nuclei of some deteriorated cells showed clear signs of pyknosis and intertubular extensive hemorrhagic areas were also observed.

The damaging effects of the organophos-phorous insecticides on the kidney were no-ticed by many investigators. In this respect, Mikhail et al. (1979) reported that the acute poisoning with dursban caused cloudy swell-ing of the convoluted tubules of the treated mice. Zaleska Freljan et al. (1983) noticed vacuolar degeneration of the lining of the proximal convoluted tubules in mice given bro-mfenvinphos alone and in combination with methoxychlor once a day for 6 weeks. Cura-cron caused degeneration followed by dilation of the proximal and distal convoluted tubules which contained hyaline casts as reveald by Enan et al. (1986) who studied the effect of 1/10 LD50 of curacron on 60 white male rats.

In the present study the golmerular tufts showed marked alterations including shrink-age, hypertrophy, lobulation and/or conges-tion. Also areas of intertubular hemorrhage were obvious in high and accumulative dos-es. Similar results were reported by Abdel Rahman and Zaki (1992) who noticed de-formation of the structure of the glomeruli of the cortical region in mice treated with mala-thion or sevin. Abdel Hadi and Abedin (1997) stated that mothers and newborn rats after treatment with dimethoate showed swelling of Bowman's capsules and tubular degenera-tion. Moreover, the authors suggested that the insecticides seem to affect newborn through the transmission of their chemicals through the mother's placenta. On the other hand, many teratogenic effects that have been ob-served in the kidney of rat fetuses maternally treated with fl ufenoxuron during organogen-esis by Karim (1998) included massive de-generation and necrosis of the epithelial cells lining the renal tubules. Moreover, congested glomeruli and intertubular capillaries and atro-phied lobulated glomeruli were also observed.

In the present investigation, the exami-nation of the lung of 18-days-old fetuses group II showed congestion of pulmonary

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blood vessels and capillaries and thicken-ing of the interalveolar septa with infi ltrated lymphocytic or infl ammatory cells. Marked pathological lesions were observed in group III which included vacuolated cytoplasm, py-knotic nuclei as well as destructed epithelium of the respiratory bronchioles with detached cells in their lumina were observed. In group IV the lung of 18-days-old fetuses exhibited highly congested blood vessels and capillar-ies, hyperplasia of the interalveoler walls with large number of lymphatic infl ammatory cells.

The results observed in the present study confi rm the results of some investigators who reported that the administration of organo-phosphorous insecticides caused pulmonary edema, thickening and vacuolization of the al-veolar cells, marked morphologic alterations in the pulmonary parenchyma, necrosis in Clara cells, hypertrophy and hyperplasia of type II alveolar epithelial cells and Collagen deposi-tion in the interalveolar septa (Durham and Imamura, 1988 ; Durham and Gijbels, 1989).

In addition, Pongrac et al. (1989) stated that cytomorphological changes occurred to the bronchial epithelial cells of a patient who inhaled aerosol organophosphate insecticide "Ekalux 25". The authors added that bronchi-al epithelial cells and macrophages showed degenerative changes, squamous meta-plasia and hypertropic cells of the bronchial epithelium were detected by the examina-tion of the patient's sputum. Moreover, con-gestion of pulmonary tissues and interstitial edema were reported by Lainee et al. (1991) who attributed this edema to the increase in permeability of the pulmonary capillaries.

On the other hand, the results of the pres-ent investigation contradict with those of Coulombe et al. (1986) who could not detect any permanent changes in the alveolar area of the rat lung after a single exposure to the organophosphorous insecticide fenitrothion.

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THE EFFECT OF AN ORGANOPHOSPHOROUS INSECTICIDE …

كبد وكلي ورئة أجنة الفئران عليأحد المبيدات الحشريةتأثيـر

ريھـاب عاشــوإ - حـمزة أحمـد الشـبكة -يـمان زكرياإ - فـوزي إبـراھـيم عامـر

جامعة عين شمس-كلية العلوم - قسم علم الحيوان

ملخص البحث

علي كبد ) همن مجموعه المركبات العضوية الفوسفوري(لدراسة تأثيرات أحد المبيدات الحشرية وھو الكوراكرون تناولت ھذة ا

اتخذت , فئران في كل مجموعة١٠, وقد تم تقسيم األمھات الحوامل إلى أربع مجموعات. وكلي ورئة أجنة الفئران البيضاء

اد المجموعتين الثانية والثالثة في اليوم السابع من الحمل عن طريق الفم المجموعة األولى كمجموعه ضابطة بينما تم معالجه أفر

أما المجموعة الرابعة فقد تم معالجتھا , كجم من وزن الجسم من الكوراكرون علي التوالي/ مج٢٨.٨كجم و / مجم١٤.٤بجرعات

ليوم السابع وحتى اليوم الرابع عشر من كجم من وزن الجسم من الكوراكرون لمده ثمانية أيام متتالية من ا/ مجم١٤.٤بجرعات

.الحمل

تم قتل األمھات الحوامل لكل من المجموعات المعالجة والمجموعة الضابطه في اليوم الثامن عشر من الحمل وتم استخراج

.و أعداد أعضاء الدراسة للفحص النسيجى اجنتھا

أوضحت الدراسات النسيجية لكبد االجنه في اليوم الثامن عشر من الحمل للمجموعات المعالجة المختلفه وجود تغيرات مرضيه

نسيجية مختلفة كان اشھرھا فقدان الترتيب المعروف للخاليا الكبديه التي ظھرت منتفخة باالضافه إلى التحلل الدھني مع مراحل

أما في الكلية فقد أوضح , رت االورده المركزية والجييبات متسعة وممتلئة بتجمعات دمويةمختلفة من تحلل انويتھا كما ظھ

الفحص المجھري لنسيج الكلية الجنه األمھات المعالجة بالمبيد تقلص واحتقان الكبب الكلوية مع تحلل الخاليا المبطنه لالنيبيبات

كما اظھر الفحص المجھري لنسيج الرئة , ي المناطق بين االنيبيباتالبوليه وبعض االنيبيبات المجمعه مع وجود تجمعات دموية ف

و تغلظا في جدران الحواجز بين الحويصالت مع , الدمويةهفي اجنه الفئران لألمھات المعالجة بالمبيد احتقان واتساع أالوعي

.نفسيةوجود خاليا منفصلة في تجويف الحويصالت الھوائية باالضافه إلى تدمير جدر الشعيرات الت

مما سبق يتضح أن الجرعات التراكمية للكوراكون لھا تاثير ضار على كبد و كلى ورئة اجنة الفئران و يجب على اإلناث

. الحوامل الحرص الشديد لتجنب تلك اآلثار الضارة على أجنتھا

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