The effect of cadmium on oogenesis in Xenopus laevis

Reproductive toxicity studies have historically centered on post-fertilization events. A thorough assessment of reproductive hazards to an organism should include all aspects of its life cycle. Cadmium is a teratogenic and carcinogenic heavy metal that occurs naturally in the environment but is also released anthropogenically. The effect of cadmium administration on oocyte development in Xenopus laevis was studied. Adult female Xenopus were injected in the dorsal lymph sac with cadmium chloride (CdCl2) at doses of 0.5, 0.75, 1.0, 3.0 or 5.0 mg/kg every other day for 21 days. Significant adverse effects of Cd on oocyte development were observed. The percentage of oocytes at all stages of oogenesis was decreased while the population of atretic oocytes increased dramatically (P < 0.0001). Numerous oocytes exhibited a speckled or mottled appearance and the incidence of completely atretic oocyte follicles increased. The observations indicate that Cd has the potential to significantly disrupt oogenesis and that examination of developing gametes may be a useful parameter for assessing the influence of environmental contaminants on reproductive capacity.     

Cypermethrin induced histological changes in gonadotrophic cells, liver, gonads, plasma levels of estradiol-17beta and 11-ketotestosterone, and sperm motility in Heteropneustes fossilis (Bloch)

The aim of the present investigation is to assess the impact of cypermethrin on reproductive physiology in catfish, Heteropneustes fossilis during prespawning phase. Results indicate that there is a decrease in the size of gonadotrophic cells with less granulation, pycnosis in the liver, presence of immature oocytes and atretic follicles in the ovaries and gross condensation of spermatogenic cells in testes after cypermethrin exposure at sublethal concentration. The gonado-somatic index (GSI), plasma levels of estradiol-17beta (E2) and 11-ketotestosterone (11-KT) also decreases. The motility of sperm cells is dependent on the dilution (2000 times) and duration of motility is recorded 2min maximally at 90s after post-activation. The dose 0.1 and 0.01ppm is sublethal, while 1ppm is lethal on sperm motility. Results indicate that cypermethrin causes inhibition of reproduction by acting at the hypothalamo-hypophyseal-gonadal axis as is manifest from the histological observations of gonadotrophs along with disruption of follicular wall and spermatogenic cells. Obviously such changes are responsible for decreasing the steroid hormone levels which result in decreasing scale and duration of sperm motility after 45d exposure of cypermethrin in this species.     

Metabolic effects of diazinon on the European eel

Abstract

Eels were exposed to a sublethal diazinon concentration of 0.042 mg/L for exposure times of 6, 24, 48, 72 and 96 hours. Biochemical analyses of blood composition, such as plasma glucose, total plasma cholesterol and triglycerides, plasma lactate, plasma urea and uric acid, showed significant differences between treated and control animals. Plasma glucose and lactate increased after 6 hours exposure to the insecticide. Plasma cholesterol and triglycerides content decreased during 96 hours treatment. Urea levels increased at 72 hours while uric acid content decreased significantly at 24, 72 and 96 hours exposure to the pesticide.

The observed effects of diazinon on eel metabolism suggested that the treated fish was faced with a serious metabolic crisis, and the fish looked for alternative methods of metabolism to overcome the toxic stress.     

Acute toxicity, uptake and clearance of diazinon by the European eel, Anguilla anguilla (L.).

The toxicity, accumulation, and elimination of diazinon were investigated for the european eel, Anguilla anguilla. The 24, 48, 72 and 96-h median lethal concentrations (LC50) were 0.16, 0.11, 0.09 and 0.08 mg/L, respectively. Fish exposed to sublethal concentration (0.042 mg/L) accumulated diazinon in liver and muscle tissues. Bioconcentration factors (BCF) of diazinon were 1850 in liver, and 775 in muscle over the 96-h exposure period. Upon removal from diazinon containing water the contaminated fish rapidly eliminated diazinon. The excretion rate constants of this insecticide were 0.108 h-1 for liver and 0.016 h-1 for muscle. Diazinon half-lives were 16.6 and 33.2 hours for liver and muscle, respectively.     

Endosulfan and diazinon toxicity to the freshwater rotifer Brachionus calyciflorus.

The acute toxicity of endosulfan and diazinon to the freshwater rotifer Brachionus calyciflorus was determined after 24 hours exposure to these toxicants. The mean 24 hr-LC50 values were 5.15 and 29.22 mg/L for endosulfan and diazinon respectively. Based on these results, four sublethal concentrations were chosen to determined the median lethal time (LT50) at each concentration of toxicant tested. We also used a control with the solvent (acetone). The concentration tested were 1/5, 1/4, 1/2 and 2/3th LC50 (24hr) for both pesticides. We found a decrease in the median lethal time (LT50) with increasing pesticide concentrations. The LT50 values ranged from 6.49 to 3.48 days after endosulfan treatment, and from 6.96 to 2.49 days after diazinon exposure. No effects on survival were observed in control animals exposed to the solvent.     

Ultrastructural changes in the respiratory lamellae of the catfish, Heteropneustes fossilis after sublethal exposure to malathion

Transmission electron microscopy study of the gills of Heteropneustes fossilis, exposed to 4 mg/liter of malathion (1/3 of LC50) for 24, 48, 72, and 96 h showed significant changes in its ultrastructures. Exposure to the pesticide after 24 h caused a slightly disarrayed condition in the double layered epithelial structure. Lymphatic spaces became more apparent, and a few chloride cells appeared which protruded toward the peripheral margin of the secondary lamellae. Chloride cells were exposed to the exterior by an apical pit. Pinocytosis was observed with marginal folds (MF) originating from the pillar and epithelial cells. Some vascular constrictions were also seen in the capillaries with erythrocytes. After 48 h exposure, the outer epithelial cells were stretched into a thin boundary wall and lymphatic spaces were engorged with plasma exudate. Chloride cells transversed the whole epithelium of the lamella and came into direct contact with lymphoid space and exterior to epithelial lining. Basement membrane of the capillaries became thicker. After 72 h a distorted lamellar epithelium ruptured in a few places allowing many spheroid bodies and some chloride cells come out. Marginal folds of pillar cells migrated into vascular spaces. Basement membrane of capillaries became thicker and blood channels were constricted causing vascular stasis. No erythrocytes were visible. Blood channels were filled with leukocytes and amoebocytes. After 96 h exposure to malathion narrowing of lymphatic spaces, proliferation of epithelial cells and development of pinocytotic vesicles from marginal folds of pillar cell flanges were observed. Only marginal blood channels maintained normal configuration. Vascular stasis due to thickening of the basal lamina were still evident in centrally located blood channels filled with leukocytes. Vascular stasis would likely cause a decrease in respiratory efficiency. This study has revealed that the gills of H. fossilis were affected by a sublethal dose of malathion. The ultrastructural damages to the gills were observed as early as at 24 h exposure, but the most severe damage occurred at 72 h exposure. However, signs of gill structure regeneration were seen in malathion-exposed fish after 96 h.     

Sublethal effects of diazinon on the structure of the testis of bluegill,Lepomis macrochirus: a microscopic analysis

A histopathological analysis of the testis exposed to diazinon, an organophosphorus pesticide was conducted. The testis from the control fish exhibited a normal histological structure with a round, oval or somewhat rectangular shaped with closely associated seminiferous tubules. The seminiferous tubules were surrounded by a layer of connective tissue with myoid cells, Leydig cells, blood vessels, lymph vessels and nerves. Upon exposure for 24 h, the overall structure of the testis looked disrupted and more often the sperm cells appeared to be somewhat larger, but spermatid were smaller than that of the control group. The 48 h exposure caused more dense and closely associated seminiferous tubules. The 72 h exposure induced dilation of the seminiferous tubules and their lumen. Ninety-six hours exposure induced more loose and disrupted structures. In this group (96 h), there was a significant reduction in both lumen and seminiferous tubule size in comparison with the control, 24, 48, and 72 h groups. After 2 weeks of exposure hardly any lumen was seen, which imparted more dense and compacted structures. The biometric analysis gave a highly significant increase in the diameter of the seminiferous lumen. After 48 h of exposure a significant reduction was seen in diameter in comparison with that of the control group. Seventy-two hour exposure indicated an increase in diameter, whereas 96 h exhibited another decrease. One week caused the diameter to increase whereas 2 weeks of exposure caused them to decrease again. The increase in diameter of the seminiferous tabule may be caused by weakening of the surrounding connective tissue and myoid cells, due to exposure to diazinon. The change in the diameters of the seminiferous tabules was very irregular. No apparent co-relation was found between the size of the fish, body weight and weight of the testis after they have been subjected to different exposure periods. There was significant change in the germ cells' diameters. As the germ cells are the essential first step in the process of spermatogenesis, their size reduction may hinder the production of viable spermatozoa, endangering the population dynamics. In order to find out the pathways of this pesticide effects, we propose to study the histopathology of pituitary gland a vital organ of the reproductive axis.     

Cadmium effect on the ovarian structure in earthworm Dendrobaena veneta (Rosa)

Cadmium effects on the ovary structure and oocytes were tested in earthworms Dendrobaena veneta exposed to 10 and 50 mg Cd kg(-1) in soil after 10 and 20 days of the experiment. In both experimental doses cadmium caused damage to the structure of the ovary but the effects were different in each group. At 10 mg Cd kg(-1) concentration in soil, young stages of oocytes and trophocytes were most sensitive to cadmium deleterious effects whereas somatic cells in the ovarian stroma were only slightly affected. Cadmium. at a concentration of 50 mg Cd kg(-1) in soil caused most damage in the somatic cells leading to the occurrence of unnaturally swollen elements and desmosomes destruction. At both experimental concentrations cadmium induced degenerative changes in cell nuclei and an increase in number of cell organelles (RER and Golgi complex elements) in the cytoplasm of oocytes and trophocytes. These also proved to be more active. No ultrastructural changes were manifested in oogonia. In both experimental groups degenerative changes occurred as early as after 10 days of Cd exposure.     

In-vivo interaction of some organophosphorus insecticides with different biochemical targets in white rats

A comparative study between five organophosphorus insecticides: Leptophos, EPN, Cyanofenphos, Chlorpyrifos and Diazinon, was carried out for acute oral toxicity to white rats and for their in vivo interaction at 1/10th of LD50 doses with the activity of six serum enzymes after 4 wks from oral administration. Leptophos, Chlorpyrifos and diazinon exerted significant inhibition particularly to glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), glutamyltransferase (GT) and lactate dehydrogenase (LDH). Adding ascorbic acid in the diet at 0.5% enhanced the acute oral toxicity of leptophos, chlorpyrifos and diazinon. For all the compounds, presence of ascorbic acid protected a number of the monitored serum enzymes from being inhibited except for leptophos. Ascorbic acid caused hypoglycemia with sublethal doses of leptophos, chlorpyrifos, and diazinon. The synergist piperonyl butoxide alone at 750 mg/kg dose inhibited the activity of the six serum enzymes. Presence of ascorbic acid in the diet intensified the inhibitory effect of piperonyl butoxide to all enzymes except for GOT.     

Linkage of biomarkers along levels of biological complexity in juvenile and adult diazinon fed terrestrial isopod (Porcellio scaber, Isopoda, Crustacea)

In parallel laboratory experiments, we determined the effect of a typical representative of organophosphorous pesticides, diazinon, on AChE activity, lipid, protein and glycogen content, weight change, feeding activity and mortality of juvenile and adult terrestrial isopods Porcellio scaber (Isopoda, Crustacea). Organophosphorous pesticides (OP) are among the most extensively used pesticides, which have replaced organochlorine pesticides. OPs inhibit the enzyme acetylcholinesterase (AChE), resulting in neurotoxicity. They have more widespread effects on non-target organisms than do organochlorine pesticides. The aim of this study was to link effect of diazinon on target enzyme to energy reserves and to integrated biomarker responses in juvenile and adult P. scaber. The non-observed effect concentration (NOEC) for AChE activity after diazinon exposure in two weeks toxicity study with isopods was below 5 microg/g diazinon. There was a good agreement between concentrations at which AChE and survival were affected (10 microg/g diazinon in juveniles, 100 microg/g diazinon in adults). We revealed a link among AChE activity, protein content and mortality. Glycogen and lipid content, feeding activity and weight change were not affected in two weeks diazinon exposure up to 100 microg/g diazinon. Juveniles were affected at concentrations that were an order of magnitude lower than those provoking similar effects on adults. Recommendations are made for future toxicity studies with terrestrial isopods.     

Histopathology of gills in mosquitofish,Gambusia affinis after long-term exposure to sublethal concentrations of malathion

The histopathological effects of malathion, an organophosphate pesticide, on the gill tissues in mosquitofish, Gambusia affinis, were determined by light microscope. The fish were exposed to sublethal concentrations (0.01 and 0.02 mg/L) of malathion for 10, 20 and 30 days. Histological examination of the gills treated with malathion showed a variety of histopathological effects. The gill lesions included necrosis and desquamation of secondary lamellar epithelium, lifting up of epithelium, intraepithelial oedema, fusion of adjacent secondary lamellae, haemorrhage at primary lamellae, disorganization and rupture in seconder lamellae, hypertrophy and hyperplasia of epithelial cells. These alterations were time- and dose-dependent.     

Rapid behavioural and morphological responses of hydropsychid larvae (Trichoptera, Hydropsychidae) to sublethal cadmium exposure

Larvae of Hydropsyche contubernalis and H. siltalai were exposed to sublethal cadmium concentrations of 0, 0.012, 0.16 and 10 mg/litre(-1) for 72 h. Linear logit models revealed a significant increase in the frequency and degree of damage of the anal papillae of both species with increasing Cd concentration. Hydropsyche contubernalis showed a stronger and earlier anal papillae response under Cd exposure than H. siltalai, whereas in the latter species darkening of the ventral sides of the abdomen was also observed. The lowest Cd level altered the competition behaviour of Hydropsyche contubernalis larvae after only 24 h exposure, yet no visual signs of morphological damage were detected. The exposure intruder larvae spent significantly less time trying to enter the nets of resident larvae than did unexposed intruders. In addition, both the exposed intruders and exposed residents pursued different behavioural tactics during the encounters compared to their unexposed counterparts.     

Evaluation of a Daphnia magna renewal life‐cycle test method with diazinon

Abstract

Acute and chronic toxicity tests with diazinon (diethyl 2‐isopropyl‐6‐methyl‐4‐pyrimidinyl phosphorothionate) were conducted on Daphnia magna. The 24‐hr static LC50 was 0.86 μL.L^‐1. The sublethal effects of 0.05, 0.1, 0.5, 0.75 and 1.0 ngL^‐1 of diazinon concentrations on the survival, reproduction and growth of D. magna were monitored for 21 days. The algae Nannochloris oculata (5 x 10⁵ cellsmL^‐1) was used to feed the daphnids. The parameters used to determined the effect of the pesticide on D. magna were: mean total young per female; mean brood size; days to first brood; intrinsic rate of natural increase (r); growth; and survival. Reproduction as well as survival was significantly reduced at diazinon concentrations of 0.10 ngL^‐1 and higher. The intrinsic rate of natural increase (r) decreased with increasing concentrations of diazinon. Growth, as measured by body length, was depressed significantly at 0.05 ngL^‐1 of diazinon and higher concentrations. The maximum acceptable toxicant concentration (MATC) was calculated. The chronic data was used to formulate an acute/chronic ratio.     

Malathion-induced sublethal toxicity on the intestine of cricket frog (Fejervarya limnocharis)

The effect of malathion [diethyl(dimethoxythiophosphorylthio)succinate] at sublethal concentration (0.006 ppm) on intestinal parameters of cricket frog (Fejervarya limnocharis) was studied for 24 hrs to 240 hrs of exposure and remarkable histopathological alterations were observed. The study on intestinal parameters revealed acute pathological conditions in the intestinal wall. The toxic effect became evident as the cytoplasm of the cells disintegrated and the cells became empty and vacuolated. The cell membranes were also ruptured. Degenerative changes of the absorptive surface (villi) of the intestine in the different periods of exposure were pronounced. Severe atrophic nature (necrotic mucosa) of the intestine began from 48 hrs onwards to 96 hrs of exposure.     

Malathion-induced sublethal toxicity on the intestine of cricket frog (Fejervarya limnocharis)

The effect of malathion [diethyl(dimethoxythiophosphorylthio)succinate] at sublethal concentration (0.006 ppm) on intestinal parameters of cricket frog (Fejervarya limnocharis) was studied for 24 hrs to 240 hrs of exposure and remarkable histopathological alterations were observed. The study on intestinal parameters revealed acute pathological conditions in the intestinal wall. The toxic effect became evident as the cytoplasm of the cells disintegrated and the cells became empty and vacuolated. The cell membranes were also ruptured. Degenerative changes of the absorptive surface (villi) of the intestine in the different periods of exposure were pronounced. Severe atrophic nature (necrotic mucosa) of the intestine began from 48 hrs onwards to 96 hrs of exposure.     

The risks posed by deltamethrin drift to hedgerow butterflies

The susceptibilities of Pieris rapae and P. brassicae (Lepidoptera: Pieridae) to topical treatment with deltamethrin were determined. LD(50)s continued to fall for an extended period after treatment and did not reach a clear end-point for P. rapae. This indicated a failure to excrete or metabolise the active ingredient, which may have continued to exert toxic effects throughout the insect's life cycle. Weight loss by P. brassicae larvae was detected at sublethal rates of exposure and treated insects exhibited feeding inhibition and produced smaller pupae and adults. Residual exposure bioassays over 72 h for P. brassicae exposed to deltamethrin deposits on cabbage leaves, detected toxic effects at rates as low as 1/520th of field application rate. LD(50)s again fell for an extended period after exposure. A model predicting mortality at given levels of drift indicated high levels of short-term risk to larvae exposed to rates of drift deposition recorded in the field. The potential of buffer zones to reduce the toxicological impact of spray drift in the field is discussed.     

Effect of commercial formulation of four organophosphorus insecticides on the ovaries of a freshwater teleost, Mystus vittatus (bloch)--a histological and histochemical study

The chronic effect of an exposure to a sublethal concentration of commercial formulation of 4 organophosphorus insecticides such as birlane (chlorfenvinphos), gardona (tetrachlorvinphos), phosdrin (mevinphos) and malathion on midvitellogenic ovaries of Mystus vittatus was investigated using several histological and histochemical techniques. The loss of stage II and III oocytes accompanied by a significant decline in gonadosomatic index was recorded. The vitellogenesis in treated fishes was ceased as the oocytes did not advance to further stages as evidenced by the absence of lipid yolks and the lack of a PAS positive follicular layer. The RNA content of remaining stage I oocytes of exposed fishes resembled that of the stage I oocytes of the control fishes. The absence of delta 5-3 beta-hydroxysteroid dehydrogenase suggested the lack of steroidogenesis. Reduced glucose-6-phosphate dehydrogenase activity was also observed. The significance of these observations has been discussed in light of available literature.     

Histopathological lesions in the body organs of cat-fish (Heteropneustes fossilis) following mercury intoxication

Light microscopic study of different body organs of cat-fish following exposure to HgCl2 0.2 mg/L in water for 30 days revealed that focal degeneration of liver cells and disorganization of hepatic cords occurred at places. Furthermore, centrilobular atrophy and compensatory hypertrophy of some hepatic cells were also observed. In the kidneys disintegration of renal epithelium along with displacement of nuclei, shrinkage of glomeruli, breakdown of Bowman's capsule and heavy infiltration by inflammatory cells were observed. The histopathological changes noted in the intestine included degeneration of lining epithelium, and diminution of goblet cells. Microscopic section of ovaries exhibited reduction of ooplasm leading to formation of atypical oocytes. An increase in the occurrence of atretic oocytes and interfolicular spaces was also discernible. No histopathological lesions could be detected in testes of male fish probably because of the difference in the maturity of the control and experimental groups. The pathomorphological alterations in relation to mercury toxicity in spleen were associated with the disorganization of the splenic cords resulting in the displacement of lymphatic tissue cells within the substance of splenic pulp. Marked depletion of the red pulp was noticeable.