histopathological studies on hypophysis and ovary of fresh ... - wjpps

WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

Narayanaswamy et al.

World Journal of Pharmacy and Pharmaceutical Sciences

SJIF Impact Factor 2.786

Volume 3, Issue 8, 841-862.

Research Article

ISSN 2278 – 4357

HISTOPATHOLOGICAL STUDIES ON HYPOPHYSIS AND OVARY OF FRESH WATER FISH GLOSSOGOBIUS GIURIS (HAMILTON) *S.Y. Narayanaswamy, M. Ramachandra Mohan Department of Zoology, Dayananda Sagar College of Biological Sciences, Kumaraswamy layout Bangalore. Professor and Ex. Chairman, Department of Zoology, Jnanabharathi campus, Bangalore University, Bangalore.

Article Received on 16 May 2014, Revised on 20 June 2014, Accepted on 16 July 2014

ABSTRACT Histopathology of the Hypophysis and Ovary of fish G. giuris was observed during preparatory phase, on the treatment with sublethal concentrations of Malathion ( 0.05 , 0.25 and 0.5 ppm) for 24, 48, 72 and 96 hrs intervals, histology represents a useful tool to assess the

*Correspondence for Author

degree of pollution, particularly for sub-lethal and chronic effects. In

Dr. S.Y. Narayanaswamy

this paper,

Department of Zoology, Dayananda Sagar College of Biological Sciences,

propose a standardized tool for the assessment of

histological findings which can be applied to different organs. The results of the present study showed morphological changes in the cell

Kumaraswamy layout

structure. Investigation on Hypophysis and Ovary cells showed

Bangalore

degranulation and hypertrophy of cells and nuclei, after the fishes were exposed to Malathion treatment, these changes further intensified with

0.25 ppm treatment through the appearance of extensive intercellular spaces. In higher concentration of Malathion treatment with 0.5 ppm for 24 hrs, the cells showed various stages of degranulation which were distributed sparsely. A reduction in the number and diameter of the Hypophysis and Ovary cells, which indicates possible reduction in the synthesis of hormones. Keywords: Malathion , G.giuris, Ovary and Hypophysis. INTRODUCTION The use of various agrochemicals including chemical fertilizers to boost the agricultural production for high yielding varieties of crop was started in the middle of twentieth century.

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Simultaneously the use of pesticide has increased considerably effect on organs of the organism throughout the world to control the pests in agriculture sector. India is one of the developing countries with vast population making considerable use of chemical fertilizers. Pesticides and other chemicals are used for the better production of grains and vegetables. The quest for meeting rapidly growing food needs with insufficient attention to the environmental impact of agricultural policies and practices has resulted in large scale environmental degradation through soil erosion, deforestation, drought and desertification For centuries pesticides have been used in agriculture to enhance food production by eradicating unwanted insects and controlling disease vectors (1,Prakasam et. al 2001). Among pesticides organophosphorus compounds (OPS) are commonly used as insecticides (2.Storm et. al 2000). Malathion, a commonly used organophosphate is applied for mosquito control at concentrations designed to achieve a surface exposure of 5 1 g/cm2 ( 3.De Guise et. al 2004). Fish is one of the major sources of animal proteins since fish contain less connective tissue than other animals, it is easily digestable and also contain less fat which reduces the risk of human cardiovasicular disease. With so much benefits occurring from fish as a human food source, along with the agriculture fishing industry is gaining paramounting importance. Industrial agriculture has promoted extraordinary use of chemicals in the form of pesticides and insecticides ( 4.Gayathri, S., N. Latha, and M. R. Mohan (2013). The handling and disposal of hazardous chemicals from the moment of manufacture to their ultimate disposal has become a matter of great environmental concern. While the uses of pesticides/insecticides are bringing out desirable results in controlling the pests in the agriculture sector, it also produces some undesirable aspects in the ecosystem particularly on fishes in the aquatic ecosystem. Hence, it has become more important to study the effect of various and excessively used pesticides which find their way into stream of rivers which are used to fish culture and one of the common pesticides in the organophosphates.

The effect of

organophosphorus pesticides on fishes has been studied by various workers 5. Sreenivasan and Swaminathan, 1967;6. Mohan, 1991; 7.Cook et al., (2005):8. Pugazhvendan et. al., 2009, Pesticidal effects on various body parts of fish have been studied by 9. Butter and Springer (1966). 10. Elsier (1970). 11.Bhattacharya et al., (1975), 12.Mukhopadhya et al., (1982), and 13. Akarte et.al , (1986)

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have shown the exposure of the fishes to pesticides leads to a large number disturbances in the physiological processes. Among the various organophosphorus pesticides. Malathion (0, 0 - dimethyl, S- (1, 2 dicarboxy ethyl phodophoro dithyoate] is considered to be an important pesticide in controlling the pests and insects 14. Kapur et al., (1975). 15.Sadhu

and Mukhopadhya

(1983), 16.Sahai (1989). 6.Mohan (1991), and 17. (2000),8 Pugazhvendan et al ., (2009) have reported

that malathion brings impairment of steriodogenesis and gonadal

functions.18. Shukla et al .(1984) working on Sarotherodon mossambicus have suggested that the lethal and sublethal concentrations of malathion caused significant reduction in gonado somatic index (GSI). Similar results are reported by 19. Saxena and Garge (1978) in Channa punctatus and 20. Kaur and Virk (1983) is Cyprinus carpio, and 6. Mohan (1991) in Glossogobius giuris. Considerable amount of research has been focused on the structure and functions of Teleostean pituitary ( 21. Sahai,1974; 6. Mohan ,1991; 22. Banergee et al., 1992). The comparative studies of fish pituitary have been reviewed by23. Green (1951) and 24. Prasad Rao (1969). The cellular components of the pituitary of teleost vary from species to species in relation to age and stage of maturation. (25. Belsare, 1962; 26. Van over breeke and Me Bride, 1967 and 27. Tan, 1972). In teleost pituitary, gonadotrophs control the reproductive activities ( 28.Ball and Baker, 1969; 29. Devlaming, 1974 and 30. Veda and Takashi, 1981). The gonadotrophs exhibit structural changes in relation to different periods of gonadal maturity as shown in H.fossilis by 31. Sundararaj, (1960); Mystus Sihgala and Ochoryn chus norka by 26. Von Overbreeke, 1967). Cyclic variations of gonadotropins have been reported in different periods of the year by 32 Singh (1970), and 33. Singh and Singh (1981). A clear correlation seems to exist between hormone synthesis, its release and www.wjpps.com

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cytological appearance of the cells. However, there is a paucity of information with regard to basic description and staining affinities of gonadotrophs in gobiid hypophysis. Histological study of hypophysis of the gobiid fish G. giuris during reproductive cycle has revealed that basophils exhibit changes in the ovary 34.(Rajalakshmi, 1966 and 35.Saxena

1980).

However a cytological change in the gonadotropin cells induced by various experimental sub lethal concentrations of Malathion treatment has not been

(C10H19O6PS2) reported so far. Therefore in the present investigation an attempt has been made to study the responses of Hypophysis cells and ovary to Malathion treatment and to compare the functional significance of these cells in relation to hypophysis during reproductive cycle. The pattern and distribution of prolactin secreting cells in rostral pans distalis (RPD) of the adenohypophysis have been studied in telcost. 36.(Boddingius, 1975 and 37.olivereace and Ball, 1964). Prolactin cells are mostly concerned with osmoregulation 28.Ball 1969, and 38. Schreibman et al., 1973) and hence considerable amount of work has been done for activation of these cells after transferring the fish to sea water 39.(Nagahama et al., 1973). 41. Singh and Singh (1982) suggested that malathion altered gonadotropin secretion in the hypophysis, which in turn significantly decreased the main activity during all phases of annual reproductive cycle in H. fossils. 42. Ghosh (1988) suggested that the pesticides impair the activity of hypothalamic nuclei and pituitary gland which in turn might affect the maturation of oocytes in Channa puctatus 19. (Saxena and Garg, 1978). Arrest of follicle development recrudescences and atresia of follicles were noted following treatment with

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pesticides such as fenithion or carboxyl in Channa puctatus 19. (Saxena and Garg, 1978), methyl parathion in Notopterus notopterus,43. (Kapur and Kulkarni, 1987) and malathion in Saratherodon mossambicus, by 18. Shukla et al., 1984. It is now established that pesticides affect on reproduction in all vertebrates but their mechanism of action is not clearly understood in many cases. In fishes, pesticides seem to retard gonadotropin secretion 44. (Singh and Singh, 1980 and 33. 1981) thereby causing regression of the gonads corresponding studies on other teleostean species are lacking. Hence, the present investigation, histopathological studies on the hypophysis (gonadotrophs and prolactin secreting cells) and the ovary of G. giuris has been made during preparatory phase under laboratory condition after exposing the fish to different concentration of malathion. MATERIAL AND METHODS The fresh water gobiid fish Glossogobius giuris (HAM) were randomly collected in and around Bangalore using cast and gillnets (10mm). The fishes were brought alive to the laboratory and were kept in 50 l aquaria containing aerated tap water and acclimitated in laboratory conditions for 15 days prior to using them in experiments. They were keep under natural photoperiod and room temperature of 26 ± 4 ◦C and were fed daily with earth worms. These fishes were treated with 0.1% potassium permanganate solution for 15 min to get rid of dermal infections. The large and sexually mature female fishes were used in this study. Females were identified externally by the presence of urinogenital papillae. The body length and weight of each fishes were recorded. The stock solution of 1 mg/L is prepared and the desired concentration are obtained by adopting the dilution technique outlined by 45.APHA (1995). The acclimitated fishes were divided into three experimental groups of six fishes each. The first three groups of fishes are placed in 0.05, 0.25 and 0.5 ppm of malathion for 24,48,72 and 96 hrs while fourth group kept in fresh water (control). Six fishes in each concentration in 10 l capacity glass trough. For all experiments the acclimitated fishes were starved for 24 hr prior to their exposure to malathion use in experiment and were not feed during the course of the experiment 46. (Dalela et al., 1978). The water was changed on alternate days and the concentration of pesticide was maintained. The pesticide treated fishes (24, 48, 72 and 96 hr) were sacrificed by decapitation and pituitary along with brain are fixed in different fixatives. Serial sagittal sections were cut 5 to 6 µ and stained for staining techniques as for the pituitary staining. Similarly, after studying www.wjpps.com

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the ovary in situ they were carefully extirpated, weighed and processed for further histological studies. The GSI was also calculated

RESULTS Prolactin secreting cells (PRL) of conrol fishes showed that the nuclei are either round or oval situated almost in the centre of the cells (diameter: 5.25µ). In few cells the nuclei of the granulated acidophils were pushed to one side of the cells. The cells were usually have central nuclei containing one or two nuclei. The cytoplasm exhibits homogenous granulation (Fig.1) and stains deeply.The Gonadotropin secreting cells (GTH) cells are comparatively large in shape with distinct nuclei and nucleoli. The staining reaction varies with the stage of granulation or degranulation in the cytoplasm, the nuclei diameter (6.23µ). The ovaries of G. giuri possess distinct ovocoel, each ovary enclosed in a thin layer of connective tissue and tunica albuginea, the large numbers of oocytes were distributed the GSI (5.63) and percentages of stag I oocytes (43.26%). The prolactin secreting cells of 0.05 ppm of malathion treatment for 24 hrs, the RPD exhibits cytomorphological changes. The cytoplasm is filled with granules, homogenously distributed and highly vascularised. The cells nuclei diameter slightly decreases when compared to the control (Control: 5.25µ treated5.23µ). Few vacuoles are also seen. The Gonadotrophin secreting cells (GTH) of fishes examined 24 hrs after malathion treatment with 0.05 ppm, the hypophysis contains both granulated and degranulated cells in the PPD region. Cells are vacuolated contain secretory substance with stain deeply. The nuclei are small with prominent nucleoli (nuclear diameter control:4.21, treated 3.13 µ).Treatment with 0.05 ppm of malathion for 24 hrs showed, the ovary consists of a thick germinal epithelium layer beneath the peritoneum. Few oocytes have ruptured follicles showing sign of disintegration. The GSI slightly lesser when compared to the control (GSI control: 5.63 treated 5.5). In most of the developing oocytes shows degeneration with an abundance of stage I and II oocytes.Treatment with 0.25 ppm of malathion for 24 hrs showed loosely arranged with less granular cells and also inter cellular spaces were predominant. The PRL cell and nuclear diameter decreases (2.15 µ) when compared to the control. Treatment with 0.25 ppm of malathion for 24 hrs showed, the cytoplasm of GTH cells showed degranulation with vesicular fluid. GTH cell nuclear diameter decreases to that of control (control:4.21, treated 2.98 µ) the nuclei appeared with dense nuclear membrane and prominent nucleoli. Few

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vacuoles were also present in the cytoplasm.Treatment with 0.25 ppm of malathion for 24 hrs of ovary exhibit more number of immature oocytes, these are spherical or ovoid and are found close to ovigerous lamellae. Ovarian growth was inhibited significantly (GSI: control: 4.83 µ) characterized by the presence of III and IV stages of oocytes.The fishes exposed to 0.5 ppm of malathion, the PRL cells showed the decrease in diameter (5.19 µ) few small vacuoles were found. The cells showed number of degranulated cells.When fishes kept in 0.5 ppm of malathion, the GTH cells exhibited degranulation and decrease in size of the cell and nuclei (cell: 5.43 nuclei: 2.23 µ). Slightly increases the number of vacuoles.The ovaries of 0.5 ppm malathion, oocytes showed that number of stage IV, V and VI oocytes simultaneously decreases in number and size of stage VI oocytes (GSI: 4.93). Prolactin secreting cells (PRL) cells fishes exposed to 0.05 ppm of malathion for 48 hrs, PRL cells showed that decrease in diameter (5.17µ) few small vacuoles were found and showed degranulated cells.In fishes examined 48 hrs after malathion treatment of 0.05 ppm the GTH cells exhibited vacuolization and degranulation containing distinct nuclei, which stained feebly. The nuclear diameter decreases (3.02µ) when compared to that of control. The ovaries of 0.05 ppm malathion for 48 hrs treatment, the oocytes showed decrease in number and size of stage IV and V oocytes. Few growing oocytes and atretic oocytes were also observed. 48 hrs of 0.25 ppm malathion treatment brought about significant nuclear hypertrophy (2.11µ) and degranulation of the prolactin cells in the RPD. Degranulation appears in the cytoplasmic region. When fishes were treated with 0.25 ppm of malathion for 48 hrs showed degranulation and vacuolization. These changes suggested hypersecretory of the gonadotrophin cells. The fishes treated with 0.25 ppm of malathion for 48 hrs, oocytes were vitellogenic and few yolky oocytes. Atretic oocytes formed a significant proportion of ovary. Decrease in the number and size of stage IV and V oocytes (GSI: 3.23). After malathion treatment with 0.5 ppm for 48 hrs, the PRL cells cytoplasm were undergone degranulation (Fig.13) which resulted vacuolization and inter cellular spaces. Gonadotrophin secreting cells (GTH) of Malathion treatment of 0.5 ppm for 48 hrs containing granulated and degranulated cells, number of vacuoles increases, cell and nuclear diameter lesser (3.05µ) when compared to that of control.Exposure of fresh water fish G. giuris to 0.5 ppm of malathion for 48 hrs, exhibited large number of vitellogenic oocytes and few yolky oocytes. Atretic oocytes formed and few degenerating oocytes were observed. Prolactin secreting cells (PRL) after 0.05ppm malathion treatment for 72 hrs, there was reduction in the cellular and nuclear diameter . The nuclei stained deeply as a result of accumulated of neurosecretory granules.

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Some of the prolactin cells showed degranulation.Gonadotrophin secreting cells(GTH),the fishes exposure to 0.05 ppm malathion for 72 hrs, resulted the decreases in diameter. Degranulation were predominantly observed in the cytoplasm.Treatment with 0.05 ppm for the 72 hrs of malathion, the ovaries exhibited atretic mature oocytes. The ovary showed gradually regressed in size (GSI:2.37).After malathion treatment for 72 hrs of 0.25 ppm, the PRL cells reduces in the cellular and nuclear diameter and there was enhancement in the vacuolization.When fishes are treated with 0.25 ppm malathion for 72 hrs gonadotrophs became smaller containing both granulated and degranulated cells.Treatment with 0.25 ppm for 72 hrs of malathion the ovaries exhibited shrunken follicles and lost its shape and became atresia. The oocytes were loosely arranged, decrease the size and number of oocytes (GSI: 2.37 µ). Prolactin secreting cells (PRL) after 72 hrs of treatment with 0.5 ppm malathion, showed degranulation and granulation. The mean and nuclear diameter decreases (2.04µ) when compared to that of control. Gonadotrophin secreting cells(GTH) of the fresh water fish G. giuris exposed to 0.5 ppm for 72 hrs, the GTH cells exhibited vacuolization and degranulation containing distinct nuclei, which stained feebly with cAHP. Exposure of fishes to 0.5 ppm malathion for 72 hrs, the ovary weight decreased (GSI : 2.08 µ) when compared to that of control. Histology of ovary revealed atresia of yolky oocytes and decreases in the number and size of stage I and II oocytes Prolactin secreting cells (PRL) of fishes exposed to 0.05 ppm malathion for 96 hrs, cells were spherical and brought about significant nuclear hypertrophy (2.34µ) and degranulation (fig.19) in the PRL cells of RPD. Nuclei and chromatin were distinct. Gonadotrophin secreting cells (GTH) of Fishes were treatd with 0.05 ppm malathion for 96 hrs GTH cells were moderately granulated indicating an inhibition of hormone release. Blood capillaries were also found towards the peripheral region of the gland. Treatment with 0.05 ppm malathion for 96 hrs, ovaries encased by a thin layer of peritoneum. The germinal epithelium became very thick. The ovaries were in the immature condition having a large number of stage II, III and fewer stage of I oocytes. In the fishes treated with 0.25 ppm of malathion for 96 hrs, there was reduction in the nuclear diameters (1.98µ). The average number of degranulated cells had increased towards the peripheral region of the PPD. When fishes were treated with 0.25 ppm malathion for 96 hrs, GTH cells showed granulation. Intercellular spaces and vacuoles were found. www.wjpps.com

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Plate - 1 Fig 1-6: Photomicrograph paraffin sections of pituitary gland and ovary of fresh water fish Glossogobius giuris Fig.1: T.S of control of prepatory phase Rostral pars distalis indicating Granulated cells (GC) and vacuolated cells (V). CAHP x 500 Fig.2: Section of control Proximal pars distalis showing Granulated (GC) and Nucleoli (NI).CAHP x 500 Fig.3: Section of control ovary indicating stage I & II oocytes . . Ehrlich’s haematoxylin and eosin x 200. Fig.4: Section of Rostral pars distalis,0.05 ppm malathion for 24 hrs exhibiting Granulated cells (GC) and vascularised cells (arrows). CAHP x 500 Fig.5: Section of Proximal pars distalis,0.05 ppm malathion for 24 hrs showing Granulated cells(GC) and Blood vessels (BV). CAHP x500 Fig.6: Section of ovary,0.05 ppm malathion for 24 hrs indicating stage I and II oocytes. Ehrlich’s haematoxylin and eosin x 200.

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Plate-2 Fig 7-12: Photomicrograph paraffin sections of pituitary gland and ovary of fresh water fish Glossogobius giuris Fig. 7: Section of Rostral pars distalis,0.05ppm malathion 48 hrs exihibiting Vacuoles (V) and degranulated cells (DGC). CAHP x 500 Fig. 8: Section of Proximal pars distalis 0.05 ppm malathion for 48 hrs showing degranulated Cells(DGC) and vacuoles (V). CAHP x 500 Fig. 9: Section of ovary, 0.05ppm malathion for 48 hrs showing IV and V stage oocytes Ehrich’s haematoxylin and eosin x 200 Fig. 10: Section of Rostral pars distalis,0.5 ppm malathion for 48 hrs indicating degranulated cells (DGC) and vacuoles (V). CAHP x 500 Fig. 11: Section of Proximal pars distalis,0.5 ppm malathion for 48 hrs showing vacuolization (V) degranulated cells (DGC) . CAHP x500 Fig. 12: Section of ovary,0.5 ppm malathion for48 hrs indicating IV and Atretic follicles(AF) Ehrlich’s haematoxylin and eosin x 200.

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Plate-3 Fig 13-18: Photomicrograph paraffin sections of pituitary gland and ovary of fresh water fish Glossogobius giuris Fig. 13: Section of Rostral pars distalis,0.05 ppm malathion 72 hrs exihibiting granulated cells (DGC) and intercellular spaces (arrows). CAHP x 500 Fig.14: Section of Proximal pars distalis 0.05 ppm malathion for 72 hrs showing Vacuolated (V) cells and degranulated (DGC) cells. CAHPx500 Fig. 15: Section of ovary, 0.05 ppm malathion for 72 hrs showing yolky oocytes (YO) and Immature oocytes(IM). Ehrich’s haematoxylin and eosin x 200. Fig. 16: Section of Rostral pars distalis,0.5 ppm malathion for 72 hrs indicating Vacuoles(V) and degranulated cells (DGC) . CAHP x 500 Fig. 17: Section of Proximal pars distalis,0.5 ppm malathion for 72 hrs showing Vacuoles(V) and degranulated cells (DGC) . CAHP x500 Fig. 18: Section of ovary,0.5 ppm malathion for 72 hrs indicating atretic oocytes (arrows). Ehrich’s haematoxylin and eosin x 200.

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Plate-4 Fig 19-24: Photomicrograph paraffin sections of pituitary gland and ovary of fresh water fish Glossogobius giuris Fig. 19: Section of Rostral pars distalis,0.05 ppm malathion 96 hrs exhibiting degranulated (DGC) and Vacuoles(V). CAHP x 500. Fig.20: Section of Proximal pars distalis 0.05 ppm malathion for 96 hrs showing Vacuolization (V) and degranulated (DGC) cells. CAHPx500. Fig. 21: Section of ovary, 0.05 ppm malathion for 96 hrs showing yolky oocytes (arrows) and I and II stage oocytes. Ehrlich’s haematoxylin and eosin x 200. Fig. 22: Section of Rostral pars distalis,0.5 ppm malathion for 96 hrs indicating Nucleoli (NI) degranulated cells (DGC) . CAHP x 500. Fig. 23: Section of Proximal pars distalis,0.5 ppm malathion for 96 hrs exihibiting granulated ( GC) cells and Blood vessels. CAHP x 500. Fig. 24: Section of ovary,0.5 ppm malathion for 96 hrs indicating, Yolky oocytes (YO) I, II and III stage oocytes. Ehrlich’s haematoxylin and eosin x 200 www.wjpps.com

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The ovaries of 0.25 ppm malathion solution treatement for 96 hrs, the oocytes showed degeneration of stage II and stage III and also presence of atretic follicles of stage IV and few of stage V oocytes. On treatment of fish with 0.5 ppm malathion for 96 hrs, PRL cells possessed distinct nuclei with darkly staining chromatin and nucleoli. The cells did not exhibit definite boundaries and appeared closely packed together. It also brought about significant nuclear hypertrophy (1.84µ) and degranulation in the RPD, which indicating an inhibition of hormone release. Exposure of fishes to 0.5 ppm for 96 hrs, GTH cells showed indistinct cells boundaries, cells were smaller (4.68µ) spherical or ovoid with nuclear membrane with deeply stained. Along with granulated cells and degranulated cells appeared to have increased. The ovarian growth was inhibited with malathion treatment 0.5 for 96 hrs. Some of the stages II and III oocytes exhibited degeneration. The GSI (control:5.63, treated 2.00) was less when compared to control. Oocytes were characterized by atretic changes like cytoplasmic liquification, swelling of the follicular wall and break down of nuclear membrane. DISCUSSION The morphology and anatomy of the teleostean pituitary have been well documented in many species 47.(Pickford and Atz, 1957; 48. Sundararaj, 1959 and 37. Oliverau and Bal, 1964). The histophysiology of the hypophysis of a few teleost have also been studied in detail ( 49. Leatherland, 1972; 50. Kaur and Vollrath, 1974; 51. Baker et al ., 1974 and 52. Bage et al , 1974). Eight distinct types of hormone cells were observed in the adenohypophysis of G. giuris, based on their difference in size, staining intensity and their stage of secretory activity. These results shown a correlation between the changes in the ovaries and in pituitary of G. giuris . Simultaneous changes in the basophils of PPD and acidophils of RPD, and ovary during various phases of reproductive cycle were observed. The teleostean adenohypophysis secrets a variety of hormones viz., prolactin, growth hormone, gonadotropin, thyrotropin, adrenocorticotropin and melanophore stimulating hormones (47. Pickford and Atz, 1957 ; 28. Ball, 1969 ). In the present investigations of G. giuris, found that prolactin and gonadotropin cells show marked cytological changes. ln the hypophysis of G. giuris granulated basophils (GTH) remained low when the ovarian weight had increased. Based on this presumed that the granulated basophils are responsible for the growth and development of oocytes in the ovary. However, when the development of new oocytes

in progress maximum

degranulations and vacuolization occurs in the basophils. In Tilapia 53. (Hyder, 1970b) the www.wjpps.com

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FSH like hormone seemed to be during gametogenesis. 49. Sundararaj (1959) stressed that the degranulations of these basophils of H. fossilis during preparatory period is probably associated with the release of large quantities of FSH and traces of LH for the development of the smaller oocytes situated towards the periphery. However,54. Rai (1966a) reported that the intense degranulation and vacuolization of cyanophils coincided with the onset of vitellogenesis and ovulation in Tor tor (Barbus). In G. giuris the increase in the ovarian weight may be due to the rapid development of oogonia and their subsequent transformation to stage I oocytes. In the present study on G. giuris it has been observed that the rapid release of hormones from the degranulated basophils was associated with the maturation of gonads and spawning behaviour of the fish. Similar reports have been given by 56. Raizada (1973) in Rasbora daniconius. In G. giuris several basophil cells showed extensive degranulation and became chromophobic. This indicated the release

of gonadotropin secreting cells in the

hypophysis.The GTH cells exhibited maximum vacuolization. This coincided with increase in the number of atretic follicles and oogonial proliferations in the ovary. This phenomenon suggests that lower content of gonadotropin in the basophils, with a simultaneous increase in the circulation is responsible for the development of oocytes in G. giuris. 54.Rai (1966a) who worked on Tor tor (barbus) also reported that extensive depletion of cyanophilis were accompanied by an increase in the number of atretic follicles and oogonial proliferations in the ovaries.35. Saxena (1980) suggested that the resumption of the degranualtion and degeneration process in the cytoplasm of the basophils take place during the late post spawning period in G. giuris. Similar degenerating changes have been described in the pituitary of Salmo gairdnerii 57.(Robertson and Wexler, 1962a) . The decrease in the amount of gonadotropin in the hypophysis of the spent fish resulted in the formation of corpora atretica. During the preparatory period, the prolactin cells in the RPD of G. giuris (maintained in freshwater) showed spherical shape and contained a mass of homogenous secretory material in the cytoplasm. Similar observations have been made on the prolactin cells of H. fossilis 51.(Baker et al ., 1974) and C. batrachus 58.(Jayashree and. Srinivasachar. 1980). In 24-96 hrs treated fish prolactin cells showed degranulation and vacuolization in the cytoplasm and conspicuous intercellular spaces. These cellular disturbances in the RPD may be directly related to the increase in concentration of malathion. This suggests an acute stress

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response, since prolactin is known to be released under stress condition. Similar observations have been made by 40.Jagadeesh and Sahai, (1986) in M. vittatus .59. 1988 in Anabas testudineus) and H. fossilis. 60.Monica (2005) in Rasbora daniconius,61. Karuppasamy (2000) in Channa punctatus. 62.Singh and Singh (1983) have shown that during preparatory period gonadotropic contents of pituitary gland decreased significantly when exposed to pesticides. The histological changes in the gonadotrophs of malathion exposed to G. giuris are comparable to the changes described in the gonadotrophs of teleost H. fossilis, 63(Jagadeesh and Sahai, 1989). In the present study, after 24 to 96 hrs of treatment with malathion the pituitary cells showed degranulations and hypotrophy of cells and nuclei. The cells depicted deformed shapes and vacuolization in the cytoplasm along with intercellular spaces, which might be due to the cytoplasmic disintegrations. Further, these intercellular spaces increased with the increase in the concentration of malathion. Such abnormalities in the PPD have been recorded in other fishes also 64(Vajpai and Mathur 1986 ; 65&59 Jagadeesh and Sahai, 1986 and 1988). 66 Ram and Sathyanesan (1983) also have made similar observations in C. punctatus following the mercuric chloride treatment. These changes may be the result of an altered or impaired synthesis of gonadotropin in response to the malathion. Thus in G. giuris, exposed to malathion for 24 to 96 hrs, inhibition of ovarian growth and comparable changes in the pituitary gland cells were noticed indicating a possible impairment of pituitary gonadal axis. During preparatory period the most important indication of the ovarian damage due to exposure to malathion is the reduction in the weight of the ovaries. Similar observations have been made in a few species of teleost in relation to different concentration of pesticides 19(Saxena and Garg, 1978 ; 23 Kaur Virk, 1983 and18 Shukla et al ., 1984). In G. giuris it was observed that the malathion greatly affect the secondary growth of primary oocytes due to the impairment of vitellogenesis. Such abnormalities in the ovary have also been recorded in other fishes also 16(Sahai 1989 ; 43Koppar and Kulkarni 1987, 67 Singh and Singh 1987, 68.Ghosh et al., 1999). Further 69 Saxena and Agarwal (1986) have shown that cadmium chloride blocked all the oogonal activity at the vitellogenic growth phase in Clarias batrachus and suggested that it might be due to the retardation of oocyte proliferation's, growth of oocytes and increase in the number of atretic follicles. The present study indicated that malathion caused similar nature of ovarian structure during preparatory period. This may be due to changes in synthesis and release of gonadotropin as a result of malathion treatment. 66 Ram and Sathyanesan (1983) pointed out the inhibition of gonadal growth and

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cytomorphological changes in the pituitary gonadotrophs of C. punctatus after exposure to mercury. This suggests the impairment of pituitary - gonadal axis without any obvious sign of gonadal degeneration. In G. giuris observed that the rate of oocyte degeneration in the ovary was more in higher concentration of malathion (0.05 to 0.5 ppm) when exposed for 24 to 96 hrs. SUMMARY AND CONCLUSION Glossogobius giuris (HAM) is a bottom dweller, distributed widely in the freshwater ponds and tanks in and around Bangalore. The present study deals with the morphological and structural changes in the hypophysis and the ovary of G. giuris in response to annual reproductive cycle, exposure of the fishes to sublethal concentration of malathion. The annual reproductive cycle of the gobiid fish G. giuris is divided into preparatory phase (April to May), pre-spawning phase (June-August), spawning phase (September to December) and post-spawning phase (January to February). The histomorpholgical aspects of hypophyseal-ovarian system of G. giuris were studied in different stages of reproductive cycle by using different staining techniques. The hypophysis is of the leptobasal type, and is broadly composed of a nervous part ( neurohypophysis)

and

glandular

part(

adenohypophysis).

Histologically,

the

adenohypophysis is divided into pro-(rostral pars distalis - RPD), meso-(proximal pars disalis - PPD) and meta - (pars intermedia - PI) adenohypophysis. Though the hypophysis is divided into three different regions, no distinct boundaries can be seen between them. Therefore, a slight mixing up of the various cell types of adjoining regions is prevalent during sexual maturity and spawning periods. The effect of malathion on the hypophyseal-ovarian axis of G. giuris has also been studied for short-term exposure (24, 48, 72 and 96 hrs to sub lethal concentration, of 0.05, 0.25 and 0.5 ppm) for histological studies. The results indicate that, malathion is capable of inhibiting ovarian development and inducing cytomorphological changes, like cytoplasmic liquification in the oocytes, deformities in nucleoli and formation of atretic oocytes. A low dose of malathion brings about a significant reduction only in the gonadosomatic index (GSI) but not in the number of gonadotrophs indicating a possible reduction in the release of

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gonadotrophins. A higher concentration of malathion reduces the GSI and also the number of gonadotrophs in the PPD, implying a possible reduction both in the synthesis and release of gonadotropins. It is inferred that, the malathion exercises an inhibitory role of the gonadal hormones, via., brain-pituitary complex. REFERENCES 1.Prakasama.A., S. Sethupathy and S. Lalitha. Plasma and RBCs antioxidant status in occupational male pesticide sprayers. Clin. Chim. Acta., 2001; 310: 107-112 2.Strom. J.E., K.R.Karl. J.Doull. Occupational exposure limits for 30 organophosphate pesticides based on inhibition of red blood cell acetylcholinesterase. Toxicol., 2000;150: 1-29. 3.Deguise. S.J. Maratea and C. Perkins. Malathion immuno toxicity in the americal Lobster (Homarus Americans) upon experimental exposure. Aquatic Toxicol. 2004; 66(4): 419425. 4.Gayathri.S., N. Latha, and M. R. Mohan. Impact of Physico-chemical Characteristics on Phytoplankton Diversity of Nalligudda Lake, Bangalore."Journal of Academia and Industrial Research (JAIR) 2.6 (2013): 349-352. 5.Sreenivasan.

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plasma proftiles of three sex hormones during different

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