Chronic stress induced duration dependent alterations in immune system and their reversibility in rats.

The objective was to find out whether severity of stress effects on immunity increases with duration of exposure and recovery depends on duration of exposure. Adult male rats (n = 30) were subjected to restraint (1 h) followed by forced swimming exercise (15 min) after a gap of 4 h daily for 2, 4 and 8 weeks and allowed to recover for 6 weeks after each exposure period. Exposure of rats to stress resulted in duration dependent significant decreases in leukocyte count, phagocytic indices of neutrophils, number of bone marrow stem cells and serum levels of IL-12 and increases in apoptotic index of peripheral blood mononuclear cells and serum levels of IL-10. The alterations in counts of neutrophils, total immunoglobulin content, phagocytic index, apoptotic index of peripheral blood mononuclear cells and serum levels of IL-10 returned to control levels in recovery group rats of 2 and 4 weeks exposure but not in that of 8 weeks exposure. However, alterations in number and apoptotic index of bone marrow stem cells returned to control levels in 2, 4 and 8 weeks stress recovery groups. The results for the first time reveal that increase in duration of exposure results in more severe damage in immune system and that shorter the exposure period, faster the recovery. In addition, in vitro study for the first time showed that corticosterone causes apoptosis of peripheral blood mononuclear cells and bone marrow stem cells in dose dependent manner. Hence death of leukocytes and their stem cells is the major cause of stress induced immune dysfunction.

Chronic stress effects and their reversibility on the Fallopian tubes and uterus in rats.

The durational effects of chronic stress on the Fallopian tubes and uterus were studied by exposing rats to stressors in the form of restraint (1h) and forced swimming (15min) daily for 4, 8 or 12 weeks. One group of stressed rats from each time period was then maintained without exposure to stressors for a further 4 weeks to assess their ability to recover from stress. All time periods of stress exposure resulted in decreased weight of the body and Fallopian tubes; however, the relative weight of the uterus and serum concentrations of oestradiol and insulin increased significantly. The antioxidant potential was decreased with increased malondialdehyde concentrations in the Fallopian tubes following all durations of exposure and after 4 and 8 weeks of stress exposure in the uterus. Interestingly, rats stressed for 12 weeks showed an increase in serum testosterone concentration and antioxidant enzyme activities with a decrease in malondialdehyde concentration in the uterus. The antioxidant enzyme activities and malondialdehyde concentration in the Fallopian tubes of all recovery group rats were similar to stressed rats. However, in the uterus these parameters were similar to controls in recovery group rats after 4 weeks or 8 weeks of exposure, but after 12 weeks of stress exposure these parameters did not return to control levels following the recovery period. These results reveal, for the first time, that chronic stress elicits an irreversible decrease in antioxidant defence in the Fallopian tubes irrespective of exposure duration, whereas the uterus develops reversible oxidative stress under short-term exposure but increased antioxidant potential with endometrial proliferation following long-term exposure.

The protective effect of the Vacha rhizome extract on chronic stress-induced immunodeficiency in rat.

CONTEXT: Chronic stress is an inevitable factor in the modern day society which affects cell mediated as well as humoral immunity. There is a need to prevent stress effects with traditionally used herbs. OBJECTIVE: The present study was undertaken to investigate the immunoprotective effect of Vacha (Acorus calamus L. Acoraceae) rhizome under stressful condition. MATERIALS AND METHODS: Soxlet extraction of Vacha rhizome was performed with increasing polarity of solvents, i.e., petroleum ether to ethanol. The extract was concentrated by distilling off the solvent in flash evaporator and dried in desiccators. The benzene extract was found to have anti-stress property in our earlier studies and hence it was used in the present experiment. Extract was administered every day for 4 weeks orally to adult female rats prior to exposure to stress, restraint (1 h) and forced swimming exercise (15 min). RESULTS: Vacha rhizome extract significantly prevented the stress induced reduction in total and differential leukocytes count, immunoglobulin content, bone marrow cellularity and viability, lymphocytes counts in lymphoid organs, islands of white pulp of spleen (ED50 = 10 mg, p < 0.001) and a significant increase in circulating immune complexes and apoptotic index of lymphoid organs (ED50 = 10 mg, p < 0.001) compared to controls. DISCUSSION AND CONCLUSION: The present study clearly indicates that Vacha extract not only prevents stress-induced suppression of immunity and structural involution of lymphoid organs, but also boosts immunity in normal rats. Therefore, it is suggested that Vacha extract administration maintains normal immunity despite the body experiencing stress.

Long-term chronic stress exposure induces PCO phenotype in rat.

Thus far the effects of chronic stress on the ovary were studied for shorter durations. However, responses of the ovary may vary with durations of exposure to stress. Hence, we investigated the responses of the ovary following exposure to different durations of chronic stress. Exposure of rats to restraint (1 h) and after a gap of 4 h to forced swimming (15 min) daily for 4 or 8 weeks resulted in significant decreases in the activities of the ovarian antioxidant enzymes, 3ß-hydroxysteroid dehydrogenase and percentage of healthy granulosa cells with concomitant increases in the number of atretic follicles, the percentage of apoptotic granulosa cells and ovarian malondialdehyde concentration. However, the response of the ovary to similar stress regime for 12 weeks was paradoxical as there were increases in the activities of ovarian antioxidant enzymes and 3ß-hydroxysteroid dehydrogenase, the number of healthy antral follicles, and decreases in ovarian malondialdehyde concentration and percentage of apoptotic granulosa cells. These changes were accompanied by hyperglycaemia and an increase in the serum levels of insulin, testosterone and oestradiol. In addition the cystic follicles were found in the ovaries of these rats. However, the number of oestrous cycles and active corpora lutea showed significant decrease in all the durations of stress exposure. The results demonstrate a differential response of ovary to short- and long-term exposure to chronic stress.

Chronic intermittent stress-induced alterations in the spermatogenesis and antioxidant status of the testis are irreversible in albino rat.

The study was undertaken to find out whether or not chronic stress-induced alterations in spermatogenesis are accompanied by oxidative damage in the testis and reversibility of these effects. Adult male rats (n = 10) were subjected to restraint for 1 h and later after a gap of 4 h to forced swimming exercise for 15 min daily for 60 days and controls (n = 5) were maintained without disturbance. After treatment period, controls and 5 rats in stress group were killed and remaining rats in stress group were maintained without any treatment for 4 months and then autopsied to find out whether effects are reversible or not. The body and testicular weight, total sperm count, and mean number of type A spermatogonia, mid-pachytene spermatocytes, stage 7 spermatids, and elongated spermatids (cellular association in stage VII of spermatogenesis) showed a significant decrease whereas the abnormal sperm count and germ cell apoptosis were increased in stressed and recovery group rats compared to controls. Activities of testicular SOD, CAT, GPx, and GST were significantly decreased whereas MDA levels were significantly increased in stressed rats compared to controls. The SOD, GST, and CAT activities of recovery groups were significantly lower than controls, whereas MDA levels and GPx activity of these rats did not differ from controls. The results, for the first time, reveal that stress-induced loss of germ cells leading to decrease in sperm count may be due to oxidative damage caused by chronic stress and majority of these changes are not reversible.

Chronic stress-induced oxidative damage and hyperlipidemia are accompanied by atherosclerotic development in rats.

Although stress-induced hyperlipidemia and increased oxidative stress have been reported and implicated in etiology of atherosclerosis, experimental evidence for stress-induced atherosclerotic development concomitant with these alterations is lacking. In this study, exposure of adult male albino Wistar rats (Rattus norvegicus) to restraint for 1 h and after a gap of 4 h to forced swimming for 15 min every day for 2, 4, or 24 weeks resulted in a duration of exposure-dependent hyperlipidemia as shown by significant increases in concentrations of blood cholesterol, low-density lipoproteins, and triglycerides and decrease in high-density lipoprotein concomitant with increased oxidative stress as indicated by decrease in hepatic antioxidant enzyme activities and increase in lipid peroxidation in the liver, kidney, and heart. These alterations were accompanied by development of fibrous layer, formation of foam cells, reduction in elastic fibers, and accumulation of Oil-Red-O-positive lipid droplets in the intima of thoracic aorta following 24 weeks of stress exposure, but not after 4 weeks. The study demonstrates for the first time that (i) chronic stress-induced hyperlipidemia and oxidative damage are coupled with atherosclerotic development in rats fed with normal diet and (ii) chronic stress effects prevail even after the cessation of stress exposure as indicated by high concentration of blood cholesterol and reduced hepatic superoxide dismutase activity 20 weeks after 2 or 4 weeks of stress. This study exemplifies long-term allostatic regulation leading to a pathological state, with long-term hyperlipidemia and oxidative damage from chronic stress resulting in atherosclerosis.

Effect of methomyl formulation, a carbamate pesticide on ovarian follicular development and fertility in albino mice.

Administration of 5 mg methomyl (40%) kg(-1) b.wt. mouse(-1) (equal to 50% of LD50 dose), every day for 90 days to adult female mice resulted in a significant decrease in body weight, relative weight of the ovary, uterus and fallopian tube; mean number of small, preantral, antral and pre-ovulatory follicles and fertility compared to controls. On the other hand, total duration of the estrous cycle was significantly increased compared to controls. One month after the cessation of the treatment (a commercial methomyl preparation-lannate) the effect on estrous cycle and organ weight was not restored. Treatment of 2.5 mg or 1 mg lannate kg(-1) b. wt., although did not alter duration of the estrous cycle; relative weight of the ovary, uterus, and fallopian tube and fertility, caused a significant decrease in mean number of small follicles compared to controls. All the groups of mice treated with lannate showed loss in body weight (15.15% in 1 mg, 6.61% in 2 mg and 12.16% in 5 mg treated groups) whereas controls showed a gain in body weight (20.02%) during the period of experimentation. The results indicate that 5 mg lannate kg(-1) b. wt causes loss of follicles and infertility, whereas lower dosages (2.5 and 1 mg) reduce the number of small follicles which might shorten reproductive life span of mice.

Chronic stress and carbohydrate metabolism: persistent changes and slow return to normalcy in male albino rats.

The present study tested the hypothesis that long-term repeated exposure to stressors results in irreversible changes in carbohydrate metabolism. Groups of adult male rats (five per group) were restrained for 1 h and 4 h later were forced to swim for 15 min everyday for 2, 4, or 24 weeks; five rats were autopsied after each interval. Groups of five rats exposed to stress for 2 or 4 weeks were maintained without further treatment (recovery groups) for up to 24 weeks. The fasting blood glucose concentration, measured at weekly intervals, was significantly higher in the stressed rats than in controls throughout the experiment, except in the 24th week, whereas that of the recovery groups was significantly higher than controls only up to the 8th week after the end of stress exposure and then reached normalcy. The blood concentrations of glucose, lactate, and pyruvate were significantly higher in the 2 and 4 weeks stress groups than in controls, whereas, except for lactate, in rats stressed for 24 weeks these values did not significantly differ from those in controls. These changes were accompanied by increased gluconeogenesis and glycogenolysis as shown by alterations in activities of hepatic carbohydrate metabolizing enzymes and unaltered blood insulin concentrations in rats stressed for 2, 4, and 24 weeks. Furthermore, the blood insulin levels did not significantly vary among controls and the 2, 4, and 24 weeks stress groups. The results reveal that though hyperglycemia induced by long-term stress exposure is reversible, it persists for a prolonged period, even after the termination of stress exposure, before reaching normalcy. Prevalence of hyperglycemia for a prolonged period through increased activities of hepatic enzymes in stressed rats exemplifies allostasis.

Progesterone prevents corticosterone mediated inhibition of estrous behaviour in rats.

Stress induced by application of electric foot shocks (300 microA/shock, five shocks per episode, 4 episodes at 1800, 1830, 1900 and 1930 hrs on the proestrus day) to rats at the time of pre-ovulatory progesterone secretion, abolished lordosis and resulted in maximum rejection co-efficient, whereas treatment with a CRF receptor antagonist (alpha-helical CRF9-41) or metapirone, an inhibitor of corticosterone synthesis, prior to application of the electric foot shocks, resulted in normal lordosis and a significant reduction in rejection coefficient. Further, administration of a single dose of corticosterone (40 microg) at 1800 hrs of proestrus caused inhibition of lordosis and resulted in maximum rejection co-efficient. On the other hand, corticosterone + progesterone treatment at 1800 hrs of proestrus resulted in normal lordosis and a significant reduction in rejection coefficient. The facts that stress induced inhibition of lordosis is prevented by CRF receptor antagonist or metapirone and that corticosterone inhibits lordosis indicate that stress induced inhibition of lordosis is mediated by corticosterone. Further, normal display of lordosis by rats treated with corticosterone + progesterone in contrast to its absence in corticosterone alone treated rats suggests that impaired progesterone secretion due to action of corticosterone leads to inhibition of lordosis.

Cypermethrin reversibly alters sperm count without altering fertility in mice.

Administration of cypermethrin (CYP), orally by gavage (3 doses: 1.38, 2.76, and 5.52 mg/kg body weight) to mice either for 6 (D1) or 12 (D2) weeks caused a significant reduction in epididymal spermatozoa count and an increase in abnormal spermatozoa count when compared to controls. These counts returned to normal levels 6 weeks after cessation of 1.38 or 2.76 mg/kg body weight (BW) treatment either after D1 or D2. In 5.52 mg/kg BW treated mice the counts returned to normal levels following D1 but not after D2. Mice in all the treatment groups showed normal fertility. Weight of the litter born to mice mated with CYP treated (all three doses) males either in D1 or D2 was significantly lower than controls whereas gestation period and litter size did not significantly vary from controls. This is the first report revealing that CYP as low as 1.38 mg/kg BW adversely affects spermatogenesis and that the effect is reversible up to 2.76 mg/kg BW/kg BW exposure for 3 months. The results further reveal that despite reduction in sperm count and increase in proportion of abnormal spermatozoa, normal fertility is possible. Hence, in reproductive toxicity evaluation of pesticides, fertility test alone is misleading.

Concentration of cations in different parts of male reproductive system and their influence on in vitro sperm motility in lizard, Mabuya carinata Schneider.

Concentrations of mono- and divalent cations in the testis, three different regions of the epididymis and the vas deferens of lizard, M. carinata were determined. The concentrations of Na+, K+ and Ca2+ increased gradually from testis to vas deferens. However, no significant variation in the levels of Mg2+ was observed between testis and anterior epididymis, and between posterior epididymis and vas deferens. Further, in an in vitro sperm motility study, wherein different concentrations of the cations were tested, the maximum forward progressive motility of posterior epididymal spermatozoa was observed following treatment with 140 mMNaCl, 25 mM KCl, and 50 mM CaCl2 whereas, treatment with magnesium brought about a decrease in sperm motility. The results indicate that Na+, K+, and Ca2+ have stimulatory effect at an optimum dose, whereas Mg2+ has an inhibitory effect even at very low concentration, on sperm motility in the lizard M carinata.

Isolation and characterization of "Reprotoxin", a novel protein complex from Daboia russelii snake venom.

In snake venoms, non-covalent protein-protein interaction leads to protein complexes with synergistic and, at times, distinct pharmacological activities. Here we describe a new protein complex containing phospholipaseA(2) (PLA(2)), protease, and a trypsin inhibitor. It is isolated from the venom of Daboia russelii by gel permeation chromatography, on a Sephadex G-75 column. This 44.6 kDa complex exhibits only phospholipase A(2) activity. In the presence of 8M urea it is well resolved into protease (29.1 kDa), PLA(2) (13 kDa), and trypsin inhibitor (6.5 kDa) peaks. The complex showed an LD(50) of 5.06 mg/kg body weight in mice. It inhibited the frequency of spontaneous release of neurotransmitter in hippocampal neurons. It also caused peritoneal bleeding, and edema in the mouse foot pads. Interestingly, the complex caused degeneration of both the germ cells and the mouse Leydig cells of mouse testis. A significant reduction in both the diameter of the seminiferous tubules and height of the seminiferous epithelia were observed following intraperitoneal injection of the sub-lethal dose (3 mg/kg body weight). This effect of the toxin is supported by the increase in the activities of acid and alkaline phosphatases and the nitric oxide content in the testes, and a decrease in the ATPase activity. Because of its potent organ atrophic effects on the reproductive organs, the toxin is named "Reprotoxin". This is the first report demonstrating toxicity to the reproductive system by a toxin isolated from snake venom.

Progesterone restores stress induced inhibition of estrous behavior in rat.

Exposure to a stressor (mild electrical shocks to foot, five times per episode, at 1800, 1830, 1900 and 1930 hrs of proestrus) coinciding with period of pre-ovulatory progesterone secretion in rats abolished estrous behavior as shown by the absence of lordosis response and a significant increase in rejection quotient compared to controls. These rats did not show spermatozoa in the vaginal smear next day morning in contrast to their presence in controls. On the other hand, rats treated with progesterone (a single injection, 500 microg in 0.1 ml olive oil at 1800 hr of proestrus) prior to exposure to stressor showed normal estrous behavior, as shown by significantly lower rejection quotient than rats exposed to stress alone, lordosis quotient similar to controls and presence of spermatozoa in the vaginal smear next day. The results, albeit indirectly, to the best of our knowledge, first time indicate that stress induced impaired steroidogenesis leads to suppression of estrous behavior.

Testis of the lizard Mabuya carinata: a light microscopic and ultrastructural seasonal study.

Histomorphology and ultrastructure of the testis during breeding and nonbreeding phases of the reproductive cycle of the lizard Mabuya carinata are studied. Observations of the ultrastructural features of the testis during breeding and nonbreeding phases of the reproductive cycle reveal a prenuptial type of spermatogenesis and a clearcut discontinuous spermatogenic cycle. Seminiferous tubules are enlarged and there is active spermatogenesis as shown by the presence of all the stages of spermatogenesis (spermatogonia to spermatids) and spermatozoa during the breeding phase (November). During the nonbreeding phase (April) only spermatogonia and Sertoli cells are seen in the shrunken seminiferous tubules. Leydig cells and Sertoli cells show distinct changes in the morphological appearance with hypertrophy of the cells in breeding phase and atrophy of the cells in the nonbreeding phase of the reproductive cycle. The present study suggests that Sertoli cells and Leydig cells functions are synchronous in the lizard M. carinata.

A study on physicochemical parameters of an aquaculture body in Mysore city, Karnataka, India.

Monthly changes in water quality parameters (physicochemical) of a rain fed lake (Bilikere) in Mysore city, were investigated for two calendar years (2002 and 2003) to assess the suitability of this lake for pisciculture. Although there were monthly fluctuations in water temperature, total suspended solids (TSS), dissolved oxygen (DO), nitrite and ammonia, they were within the desirable limits. On the other hand, total alkalinity and hydrogen sulphide throughout the study period and pH for a major part, were higher than the desirable limits. Other parameters viz; turbidity, biological oxygen demand (BOD), phosphate, and nitrate in a few months were higher than the desirable limits for waters used for fish culture. The high levels of these factors are due to the entry of agricultural run off and occasional flow of sewage into the lake. In addition dense algal growth was noticed at times of the year which is caused by surge in nutrients level whenever there was a rainfall. Since, the lake has a great aquacultural potential, it is suggested that control of nutrient load that enters the lake occasionally, might help the lake to continue its mesotrophic status.

Ultrastructural study of the epididymis and the vas deferens and electrophoretic profile of their luminal fluid proteins in the lizard Mabuya carinata.

The light microscopy, histochemical and TEM studies of the epididymis and the vas deferens revealed the presence of PAS positive secretory granules in the epithelial cells lining the lumen of these organs. One dimensional SDS gel electrophoretic pattern of luminal fluid proteins and the total protein content of the testis, three regions of the epididymis and the vas deferens of the lizard, Mabuya carinata were studied during breeding and nonbreeding season of the reproductive cycle. During breeding season, 25 protein bands in the testicular luminal fluid, 26 in the anterior epididymal luminal fluid and 28 in the middle and posterior epididymal luminal fluid were found. Ten new protein bands appeared in the anterior epididymal region whereas five new protein bands appeared in the middle region of the epididymis indicating regional difference in protein secretions of the epididymis. Vas deferens luminal fluid showed the highest number of protein bands (32) and the highest total protein content (9.07 mg/ml) compared to the testis and the epididymis. Four new protein bands appeared in the vas deferens. Number of protein bands in the luminal fluids of testis, epididymis and the vas deferens were significantly reduced during nonbreeding season compared to those of the breeding season. Consistent with the decrease in the number of protein bands, there was a significant reduction in the total protein concentration in all the tissue samples during nonbreeding season. The results indicate seasonal differences in number of proteins secreted and quantity of proteins in the luminal fluid of male reproductive tract of M. carinata. This is the first study in reptiles revealing appearance of new proteins in epididymis, and vas deferens by conducting simultaneous electrophoretic profile of testicular, epididymal and vas deferens luminal contents.

Influence of stress on gonadotrophin induced testicular recrudescence in the lizard Mabuya Carinata.

Administration (ip) of FSH (10 IU/0.1 ml distilled water (dw)/lizard/alternate days/30 days) to adult male lizards, Mabuya carinata, during the early recrudescence phase of the reproductive cycle caused activation of spermatogenic and steroidogenic activity of the testis, as shown by a significant increase in mean number of spermatogonia, primary spermatocytes and spermatids, and serum levels of testosterone, as compared to initial controls. In addition, there were abundant spermatozoa in the lumen of the seminiferous tubules. Interestingly, administration of a similar dosage of FSH to lizards exposed to stressors (handling, chasing, and noise randomly applied, five times a day for 30 days) resulted in a significant increase in mean number of spermatogonia and primary spermatocytes over initial control values, whereas the number of secondary spermatocytes and spermatids and serum levels of testosterone did not significantly differ from those of initial controls, and were significantly lower than FSH treated normal lizards. Further, spermatozoa were infrequently found in the seminiferous tubules of these lizards. Treatment controls (receiving 0.1 ml dw/lizard/alternate days for 30 days) did not show significant variation in mean number of spermatogonia, spermatocytes and spermatids, and serum levels of testosterone from initial controls. Another group of lizards was exposed to stressors and did not receive FSH. These lizards showed a significant decrease in mean number of secondary spermatocytes compared to treatment controls and all other parameters did not significantly differ from those of both control groups. The results reveal that gonadotrophin-induced spermatogonial proliferation occurs under stressful conditions, whereas progress of spermatogenesis beyond primary spermatocyte stage is impaired due to inhibition (under stress) of gonadotrophin induced steroidogenic activity in M. carinata.

Light microscopical and ultrastructural studies on the vas deferens of the lizard Mabuya carinata.

Adult male lizards (Mabuya carinata) were studied during breeding and non breeding seasons to determine the regional and seasonal differences if any in the vas deferens and to compare ultrastructural features of luminal epithelial cells with those of endotherms. The vas deferens of the lizard is a convoluted tube extending from the epididymis to the hemipenis passing over the kidney. Based on morphometric data of luminal diameter and epithelial cell height three distinct regions viz; proximal, middle and distal regions were identified in the vas deferens. The epithelium is surrounded by a thin layer of lamina propria, many layers of circular smooth muscle fibers and an outer layer of visceral pleuro peritoneum. Based on cell and nuclear morphology and ultrastructure, five different cell types viz; principal cell, basal cell, mitochondria rich cell, halo cell and narrow cell were identified in the epithelium during both breeding and non breeding season. Principal cells and basal cells were more abundant in both seasons. The types of luminal epithelial cells of vas deferens of M. carinata and their ultrastructural features are similar to those of mammals. Further, vas deferens of M. carinata differs from mammals in having only circular smooth muscles in contrast to circular and longitudinal muscles of mammalian vas deferens. To the best of our knowledge this is the first report describing cell types of vas deferens, their ultrastructure and ultrastructural seasonal variations in reptiles.

Beta-endorphin disrupts seasonal and FSH-induced ovarian recrudescence in the lizard Mabuya carinata.

Administration (ip) of an opioid peptide, beta-endorphin (beta-EP) (0.1, 0.5, or 1 microg beta-EP/day/lizard for 30 days) during seasonal recrudescence phase of the ovarian cycle inhibited ovarian recrudescence as shown by the absence of vitellogenic follicles in the ovary in contrast to their presence in treatment controls in the lizard Mabuya carinata. In the germinal bed, treatment of 0.1 microg beta-EP did not affect primordial follicles, whereas their mean number was significantly lower in lizards treated with 0.5 or 1 microg beta-EP compared to those of treatment controls. There was also suppression of oviductal development as shown by a significantly lower relative weight of the oviduct and regressed oviductal glands in lizards treated with all the dosages of beta-EP compared to treatment controls. In another experiment, administration of FSH (10 IU FSH/alternate day/lizard for 30 days) during the regression phase of the ovarian cycle induced development of vitellogenic follicles, whereas the treatment controls showed only previtellogenic follicles. In addition, there was a significant increase in the ovarian and oviductal weights compared to initial and treatment controls. However, simultaneous administration of similar dosage of FSH and beta-EP (0.5 microg/day/lizard) did not induce ovarian recrudescence as shown by the absence of vitellogenic follicles in the ovary and significantly lower weight of the ovary and the oviduct and the mean number of oogonia, oocytes, and primordial follicles compared to those of FSH-treated lizards. The results indicate that beta-EP inhibits seasonal as well as FSH-induced ovarian recrudescence. Inhibitory effect of beta-EP on follicular development despite FSH administration implies its effect at the ovarian level in M. carinata. While adversely affecting the ovarian follicular development, beta-EP did not affect the adrenal gland as there was no significant variation in the mean nuclear diameter of the adrenocortical cells of treatment controls and beta-EP-treated lizards. Furthermore, administration of beta-EP caused a significant decrease in the mean number of islands of white pulp of the spleen indicating its adverse effect on immunity.

Stress inhibits seasonal and FSH-induced ovarian recrudescence in the lizard,Mabuya carinata.

Stressors (handling, chasing, and noise) applied randomly five times per day for one month to lizards during the recrudescence phase of the ovarian cycle caused a significant reduction in mean number of oocytes and primordial follicles when compared to those of controls. Further, vitellogenic follicles were absent in the ovary of lizards subjected to stressors. Administration of bovine FSH during post-breeding regression phase of the ovarian cycle induced ovarian recrudescence as shown by significant increases in the mean number of oogonia, oocytes, and primordial follicles compared to controls, as well as vitellogenic growth of follicles. However, lizards treated with FSH and exposed to stressors did not exhibit ovarian recrudescence. Furthermore, FSH administration during the post-breeding regression phase caused a significant increase in serum levels of estradiol compared to controls, which was accompanied by significant increases in the relative weight of the liver and oviduct, as well as vitellogenic growth of follicles. Despite administration of FSH to lizards subjected to stressors, there was neither any increase in serum levels of estradiol and weight of the liver nor vitellogenic growth of follicles. The results indicate that repeated application of stressors inhibits vitellogenic growth of follicles by suppression of steroidogenic activity in M. carinata. This is the first report revealing that the ovary does not respond to gonadotrophin treatment under stressful conditions in reptiles.