Effects of sodium fluoride on hepatic toxicity in adult mice and their suckling pups

In this investigation, we have evaluated the effect of sodium fluoride (NaF) on hepatic function in pregnant and lactating mice and their suckling pups. Experiments were carried out on female Wistar mice given 500 ppm sodium fluoride (226 ppm fluoride ion) in their drinking water from the 15th day of pregnancy until day 14 after delivery. All mice were sacrificed on day 14 after parturition. Our results showed a significant decrease in serum levels of total protein and albumin, a marked hypoglycaemia and a significant decline in serum cholesterol and triglyceride levels in fluoride-treated mice and their pups. Whereas globulin and biluribin levels in serum were not significantly changed by NaF treatment. On the other hand, serum transaminase activities (aspartate transaminase; alanine transaminase), which well known as markers of liver function, were elevated indicating hepatic cells’ damage after treatment with fluoride. Lipid peroxidation increased in NaF-treated mice and pups, as revealed by high liver malondialdehyde levels, while serum total antioxidant status showed a significant decline. These biochemical modifications in NaF-treated mice also correspond histologically with extensive ballooning, hepatocellular necrosis and infiltration of mononuclear cells. These effects were not observed in controls.     

Fluoride-induced brain damages in suckling mice

In order to reveal mechanisms of brain damages resulted from fluoride toxicity, we treated adult female mice of Swiss Albinos strain by 500 ppm NaF (226 ppm F⁻) in their drinking water from the 15th day of pregnancy until the day 14 after delivery. All mice were sacrificed on day 14 after parturition. During treatment, levels of thiobarbituric acid reactive substances (TBARS), the marker of lipid peroxidation extend, increased, while the activities of the antioxidant enzymes such as glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT), and the level of glutathione (tGSH) decreased significantly in both cerebellum and cerebrum compared with those of the control group. These results suggested that fluoride enhanced oxidative stress in the brain, thereby disturbing the antioxidant defense of nursing pups. In addition, acetylcholinesterase (AChE) activity in both cerebellum and cerebrum was inhibited after treatment with fluoride. In our previous studies we have found, a decrease in plasma thyroid hormone levels of nursing pups whose mothers were treated by NaF. In cerebellum of mice, migration of neurons from the external granular layer to the internal granular layer occurred postnatally and was dependent upon the presence of thyroid hormone. In hypothyroidism, several neurons failed to complete their migration and died. Key guidance signals to these migrating neurons were provided by laminin, an extracellular matrix protein fixed to the surface of astrocytes. In the present study we examined the expression and distribution of laminin in cerebellum of 14-day-old mice. Immunoreactive laminin was disappeared by postnatal day 14 in cerebellum parenchyma of control pups and was restricted to vasculature despite the continued presence of granular cells in the external granular layer. In contrast, in cerebellum of NaF treated pups, laminin was deposited in organised punctuate clusters in the molecular layer. These data indicated that the disruption of laminin distribution might play a major role in the profound derangement of neuronal migration observed in cerebellum of NaF treated pups.     

The in vivo effect of mirex on soluble hepatic enzymes in the rat

Adult male and female rats fed dietary mirex in concentrations of 10, 20, 30, 40, and 50 ppm for 4 weeks exhibited significant decreases in liver levels of lactic dehydrogenase, malic dehydrogenase, sorbitol dehydrogenase, glutamic oxaloacetic transaminase, and glutamic pyruvic transaminase as compared to control levels. Enzyme losses were cytoplasmic and decreased in magnitude with increased time of exposure and dietary mirex concentrations. Serum sorbitol dehydrogenase levels were increased above control levels during the first week of mirex exposure while hepatic sorbitol dehydrogenase levels were concomitantly decreased below the control levels. Hepatic glutamic oxaloacetic transaminase levels were significantly decreased during the first week for all dietary mirex concentrations as were the other enzymes at the 40 and 50 ppm mirex concentrations. The magnitude of the enzyme decreases in female rat tissue was consistently lower than enzyme decreases in the male rat tissue fed equivalent dietary mirex dosages. Liver weights for male rats increased significantly at all dietary mirex levels during week one. However, only animals receiving the 40 and 50 ppm dietary mirex levels showed significant liver weight increases at the end of week 4. Female rat liver weights were increased at the 30 and 40 ppm mirex dietary levels after 4 weeks. No significant differences in body weights were observed for any dietary level of mirex. Mirex hepatic residues appeared to be equal for both sexes. No gross histological alterations were observed following treatment.     

Nile tilapia (Oreochromis niloticus), liver morphology,CYP1A activity and thyroid hormones after Endosulfan dietary exposure

Endosulfan is a worldwide used insecticide suspected to be highly toxic to aquatic organisms, including fish. Most of the available studies have focused in water exposures, although this pollutant can be transferred through food chain. Therefore, in the present study, the effects of Endosulfan on tilapia (Oreochromis niloticus), when administered through the diet. Fish were fed 21 days with diets containing 1 and 0.5 μg g⁻¹ of Endosulfan, after which qualitative histological liver analysis showed that Endosulfan induced hepatocyte destruction, vessel endothelium rupture and increased melanomacrophages aggregates. To test lower environmentally relevant doses of Endosulfan could induce hepatic damage, as well as other negative effects, such as altered phase I metabolism and plasma thyroid hormone levels. Hence, tilapia were orally exposed to 0.1 and 0.001 μg g⁻¹ for 35 days. Low environmentally realistic doses of Endosulfan were still able to induce liver histopathological damage such as increased hepatocyte vacuolization and increased eosinophil granular cell aggregates. Liver cytochrome P450 1A activity, evaluated through ethoxyresorufin-o-deethylase (EROD), was enhanced in tilapia exposed to 0.001 μg g⁻¹, whereas the highest dose had no measurable effects in this enzyme activity. Fish exposed to 0.1 μg g⁻¹ of Endosulfan had depressed T4 plasma levels. Overall, the results of the present study further demonstrate the toxic effects of Endosulfan in tilapia when administered in the diet at environmentally relevant concentrations, which indicates that in the field food chain transfer may also be an importance source of this pollutant.     

Dose-dependent sub-chronic toxicity of diethyl phthalate in female Swiss mice

The experiment was conducted to study the after effects of administering DEP at different doses to female Swiss mice for a period of 90 days. Group I mice were fed on normal diet and water ad libitum. Group II mice were maintained on normal diet mixed with corn oil at 8.25 mg/kg of the diet/day as oil control. Group III, IV and V mice were given diethyl phthalate dissolved in corn oil mixed with the diet at 10, 25 and 50 mg/kg of the diet/day, which is approximately equal to 1.25, 3.125 and 6.25 mg/kg body weight/day. A significant dose dependent increase was observed in serum acid phosphatase (ACP) whereas, serum and liver triglycerides levels showed a significant increase only in the high-dose treated group. Significant dose-dependent increase in serum aspartate and alanine aminotransferase (AST and ALT) and liver glycogen was observed. Serum lactate dehydrogenase (LDH) was significantly increased only in 25 and 50 ppm DEP-treated mice. Liver cholesterol was significantly increased in all the treated groups. Liver histology by light microscopy showed intracellular vacuolations in all the treated groups which was much more evident in the 25 and 50 ppm DEP-treated mice while hepatocellular degeneration and hypertrophy of the hepatocytes was evident in 50 ppm DEP-treated mice. Proliferation of mitochondria and peroxisomes was evident in the electron micrographs of the 10 ppm DEP-treated mice while 25 and 50 ppm DEP-treated mice showed increase in lipid droplets and severe mitochondrial proliferation.     

Evaluation of molluscicidal activities of Arisaema tubers extracts on the snail Oncomelania hupensis

Four extracts of Arisaema erubescens tubers by acetic acetal (AAE), benzinum (BZE), n-butanol (NBE) and chloroform (CFE) were obtained to evaluate their molluscicidal activities against the snail Oncomlania hupensis. The responses of choline esterase (ChE), alkaline phosphatase (ALP), esterase (EST), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) to the extracts (NBE) were also investigated. In the four extracts (AAE, BZE, NBE and CFE), NBE showed the highest toxicity on the snails after 48 h exposure. NBE also showed the time- and concentration-dependent effect, for example, the LC₉₀ values of the NBE were decreased from 365.5 mg/L (24 h) to 36.4 mg/L (96 h). At the end of exposure to NBE (LC₅₀ concentration), the activities of ChE and ALP in snail tissues (cephalopodium and liver) decreased significantly. Isozyme electrophoresis profiles indicated that responses of isozymes (EST, SOD and GSH-Px) to NBE were more intense in liver than in cephalopodium. After 72 h exposure to NBE, the EST activity in snail liver decreased and some enzyme bands (EST1 and EST4) disappeared. But the activities of SOD 1 and GSH 2 in liver increased after 48 h exposure. The results indicated that NBE was the highest toxic component in the four extracts. The decline of the detoxification ability and the oxidative damage in snail tissues might be the main reason for the molluscicidal activities.     

Effects of trichlorfon and sodium dodecyl sulphate on antioxidant defense system and acetylcholinesterase of Tilapia nilotica in vitro

The effects of organophosphorus insecticide trichlorfon, surfactant sodium dodecyl sulphate (SDS), and the mixture of trichlorfon and SDS on the antioxidant defense system and acetylcholinesterase (AChE) in Tilapia nilotica were assessed in vitro. Various concentrations of trichlorfon (0, 0.0001, 0.001, 0.01, 0.1 and 1 g/L) and SDS (0, 0.0625, 0.125, 0.25, 0.5, 1 g/L) were incubated with homogenate of liver and muscle, respectively, at 25 °C for 0, 30, 60 and 90 min. Two concentrations of mixture of trichlorfon and SDS (0.0001 g/L trichlorfon + 0.5 g/L SDS, 0.1 g/L trichlorfon + 0.5 g/L SDS) and 0.0001 g/L trichlorfon, 0.1 g/L trichlorfon, 0.5 g/L SDS and control, were incubated simultaneously with homogenate of liver and muscle, respectively, at 25 °C for 60 min. After incubation, the content of reduced-glutathione (GSH) and the activity of superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST) in homogenate of liver were determined, and the activities of AChE in homogenate of muscle were also measured.Treatment with trichlorfon caused a significant concentration-dependent and time-related inhibition of AChE activity at all treatment concentrations and times since trichlorfon is a cholinesterase inhibitor. For the same trichlorfon treatment, an apparent decrease in GSH content was found in concentration of 0.01, 0.1, 1 g/L, whereas no significant alteration in antioxidant enzyme activity were found at all experiment concentrations and times, which might indicate that antioxidant enzymes have not involved in the metabolism of trichlorfon. The depletion of GSH might indicate that ROS could be involved in the toxic effects of trichlorfon. Exposure of SDS can inhibit activities of AChE, GST and CAT at concentrations of 0.5 and/or 1 g/L, which could be due to the denaturing process of SDS to the enzymes. For the mixture exposure of trichlorfon and SDS, the effect of the mixture of 0.0001 g/L trichlorfon and 0.5 g/L SDS on inhibition of AChE shows synergistic other than simple additive of trichlorfon and SDS. The combined effects of chemicals and detergents deserve to be particularly noted. It should be noted that the toxicity experiments were made in tissue homogenates instead of whole organisms. The responses against the toxic compounds will not be the same in both systems.     

The effects of deltamethrin on some serum biochemical parameters in mice

Sixty white male mice were used in this study. Three groups, each comprising 20 mice were established. The control group (Group 1) was provided ad libitum pellet feed. On the other hand, the experimental groups, namely, Groups 2 and 3 were given pellet feed containing deltamethrin throughout the day, so that the animals were administered doses of 7.5 and 30 mg/kg/body weight/day, respectively. Blood samples were collected from all groups on the 15th, 30th, 45th, and 60th days of the experiment for measurement of serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), alcaline phosphatase (ALP), amylase and cholinesterase activities, and levels of glucose, urea, creatinine, triglyceride, cholesterol, calcium, phosphor, sodium, potassium, and chloride. According to data obtained, on the 15th day of the study, compared to the control group, statistically significant increase in urea, cholesterol, ALP, cholinesterase, calcium, and potassium levels/activities, and significant decrease in SGOT activity and chloride levels in Group 2, and significant increase in cholesterol, ALP, cholinesterase, calcium, potassium and chloride levels/activities in Group 3, and significant decrease in SGOT activity in Group 3 were determined. On the 30th day of the study, in comparison to the control group, triglyceride and cholesterol levels and ALP activity were determined to be significantly increased in the Group 2, whereas SGOT activity were significantly reduced in Group 2, and triglyceride and cholesterol levels were demonstrated to be significantly increased and SGOT activity was significantly reduced in Group 3. On the 45th day of the study, compared to the controls, significant increase in cholesterol and sodium levels, and significant decrease in triglyceride levels and ALP activity in Group 2, significant increase in creatinine, cholesterol and sodium levels and cholinesterase activity, and significant decrease in glucose, SGPT, ALP and phosphor levels/activities were detected in Group 3. On the 60th day, in other words, in the last period of the study, ALP activity, and triglyceride, calcium, and phosphor levels were determined to be significantly increased and significant decrease in amylase activity in Group 2. Furthermore, significant increase in triglyceride, ALP, amylase, calcium, phosphor, and potassium levels/activities, and significant decrease in glucose and creatinine levels were observed in Group 3. However no correlation was determined to exist between the changes were found to be statistically significant, and the administration dose and duration of deltamethrin in all periods and groups.     

Chlorpyrifos induced alterations in the levels of hydrogen peroxide,nitrate and nitrite in rat brain and liver

Organophosphate pesticides are among the most widely used synthetic chemicals for controlling a wide variety of pests. Chlorpyrifos (CPF) is among the leading organophosphate (OP) pesticides used extensively throughout the world including India. Besides being potent anticholinesterase compounds, these OP pesticides are known to generate oxidative stress. Present study was carried out to see the level of reactive oxygen species (ROS), hydrogen peroxide (H₂O₂), nitrate (NO₃⁻) and nitrite (NO₂⁻) in different parts of rat brain and liver after giving different doses of CPF intramuscularly for 3 days or for 1 month. Results of the present study clearly revealed that levels of H₂O₂, NO₃⁻ and NO₂⁻ were increased significantly in all the three parts of rat brain i.e., fore-, mid- and hind-brain as well as in liver of rats given for 3 days or 1 month due to exposure of different doses of CPF. The study clearly demonstrated that CPF generated oxidative stress in all the three brain regions as well as liver of rats and the ROS were accumulated. Accumulation of ROS in all the regions of brain and other tissues may disturb the normal physiological functions aggravating the toxicity symptoms of CPF.     

Adrenalectomy and the adaptive liver response in mirex-treated rats

Mirex, an organochlorine compound, was administered as a single oral dose (100 mg/kg body wt) to both intact and adrenalectomized juvenile male Sprague-Dawley rats. Both mirex-treated intact and adrenalectomized animals (dosed 24 hr postsurgery) exhibited significant increases in liver weight to body weight ratios compared to controls. However, the liver weight to body weight ratios in mirex-treated intact animals were significantly greater than those observed in mirextreated adrenalectomized animals. Significant increases were observed in liver weight to body weight ratios in adrenalectomized animals treated with mirex 4 days after surgery. However, the 96-hr mortality in mirex-treated adrenalectomized animals increased from 20% (mirex dose given 1 day postadrenalectomy) to 56% (mirex dose given 4 days postadrenalectomy). Mirex treatment of intact and adrenalectomized animals had no significant effect upon either serum or hepatic activities of glutamic oxalacetic transminase, glutamic pyruvic transaminase, sorbitol dehydrogenase, or protein concentrations. Bromsulfophthalein clearance also was not affected by mirex treatment in adrenalectomized animals. Serum glucose concentrations were significantly decreased in both intact and adrenalectomized animals by mirex treatment. Daily corticosterone supplements to adrenalectomized animals restored liver hypertrophy and serum glucose concentrations to levels observed in mirex-treated intact animals. These results suggest that mirex-induced liver enlargement may be mediated by corticosterone.     

Physiological and molecular mechanisms of glyphosate tolerance in an in vitro selected cotton mutant

We have selected an upland cotton (Gossypium hirsutum L.) cell line (R1098) that is highly tolerant to glyphosate. This cell line was developed by in vitro selection with gradually increasing glyphosate concentrations, and its mechanisms conferring glyphosate tolerance were studied. Based on a whole-plant dose–response bioassay, R1098 plants were tolerant to glyphosate at a concentration of 1500 g ae ha⁻¹ glyphosate (1.5× the recommended field rate) whereas the control plants (Coker 312) were unable to survive at 150 g ae ha⁻¹ glyphosate. Coker 312 accumulated 13.1 times more shikimate in leaves at 5 days after glyphosate treatment (1500 g ae ha⁻¹) than that of R1098. Two distinct cDNAs for 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), EPSPS-1 and EPSPS-2, were isolated from R1098. Both cDNAs were 97.7% identical within the common protein-coding region and the predicted sequences of the mature proteins were greater than 83% identical with EPSPS proteins from other known higher plants. In comparison to the glyphosate-susceptible cotton Coker 312, sequence analysis of the EPSPS-1 gene indicated that R1098 has an alanine insertion at nucleotide position 1216 resulting in frameshift. It leads to two copy functional EPSPS genes in R1098. There was no difference between R1098 and Coker 312 in EPSPS mRNA levels before glyphosate treatment. However, its treatment caused a 2–4 times increase in the basal EPSPS mRNA level in R1098.     

Comparative molluscicidal action of extract of Ginko biloba sarcotesta,arecoline and niclosamide on snail hosts of Schistosoma japonicum

In search for new local plant molluscicides for the control of the vectors of schistosomiasis, we compared the molluscicidal action of the extract of Ginkgo biloba sarcotesta by benzinum (EGSB) to that of arecoline (ARE) and niclosamide (NIC) against Oncomelania hupensis snails. NIC showed the highest toxicity on snails with 24 h LC₅₀ vales of 0.12 mg/L and LC₉₀ of 0.98 mg/L, while the LC₅₀ and LC₉₀ of EGSB were much lower than that of ARE. Sublethal in vivo 24 h exposure to 40% and 80% LC₅₀ of NIC, EGSB and ARE altered the activities of different enzymes in different body tissues of snails. EGSB could significantly inhibit Choline esterase (ChE), Alanine aminotransferase (ALT), Alkaline phosphatase (ALP) and Malic dehydrogenase (MDH) activities both in the cephalopodium and liver. ARE could significantly cause a reduction in ChE, ALP activities in the cephalopodium and ChE, ALT, ALP, Succinodehydrogenase (SDH), MDH activities in the liver. NIC significantly altered activities of ChE, ALT, ALP, SDH, and MDH in the cephalopodium and ChE, ALT, ALP, SDH activities in the liver. All molluscicides could not affect Lactate dehydrogenase (LDH) activity in the cephalopodium and the liver. Maximum inhibition of ALT and MDH activities was found in the cephalopodium and liver of snails treated with 80% of 24 h LC₅₀ of EGSB. However, NIC and ARE caused maximum reduction in ALP and SDH activities, respectively. The results indicated that molluscicidal action of EGSB was different to that of ARE and NIC in some extent.     

Oxygen consumption and ammonia excretion of the freshwater crab Trichodactylus borellianus exposed to chlorpyrifos and endosulfan insecticides

The effects of lethal and sublethal concentrations of chlorpyrifos and endosulfan on oxygen consumption and ammonia excretion rate of the crab Trichodactylus borellianus were evaluated. Oxygen consumption and energy expenditure had significant effect in relation to exposure times. Regarding endosulfan, a significant difference in consumption among times of exposure was registered in 625 μg L⁻¹. Moreover, at the highest concentration, energy expenditure rate was observed stabilized during 1–3 h. A significant increase in ammonia excretion was evidenced in 150 and 300 μg L⁻¹ of chlorpyrifos. The O:N ratio showed a decrease in chlorpyrifos and in 2500 μg L⁻¹ of endosulfan. This indicated a shift towards protein primary metabolism. An increment in the O:N ratio was observed in the lower endosulfan solutions. The relation oxygen:nitrogen showed a shift towards lipid and carbohydrate primary metabolism. This work indicated the complexity of the metabolism in the freshwater crab affected by xenobiotic elements.     

Evaluation of propolis effects on some biochemical parameters in rats treated with sodium fluoride

The present study in which 42 female rats, each weighing 200−250 g, were used covered a period of 21 days. The animals were divided into six groups. The first group served as the control group, whereas Group 2 was administered propolis at a dose of 200 mg/kg/bw in drinking water for 21 days. Group 3 was first provided with normal drinking water for a period of 14 days, and was subsequently administered propolis at a dose of 200 mg/kg/bw in drinking water for 7 days. Group 4 was first given normal drinking water for 14 days, and was secondly administered 100 ppm fluoride as a sodium fluoride in drinking water for 7 days. Group 5 was first administered propolis alone at a dose of 200 mg/kg/bw in drinking water for 14 days, and was secondly administered 100 ppm fluoride in association with 200 mg/kg/bw propolis for 7 days. Finally, Group 6 was first provided with normal drinking water for 14 days, and was secondly administered 100 ppm fluoride in association 200 mg/kg/bw propolis for a period of 7 days. At the end of the 21st day, blood samples were collected from the heart of each animal into both heparinised tubes and tubes without anticoagulants. Glucose, triglyceride, cholesterol, total protein, and uric acid levels, and aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) activities in the serum, as well as malondialdehyde (MDA) levels, glutathione peroxidase (GSH-Px) in the plasma, erythrocyte superoxide dismutase (SOD) and catalase (CAT) activities were measured. When compared to the control group, statistical differences were determined to exist with respect to oxidative stress parameters which involved increase in MDA levels in Groups 4−6, decrease in SOD activity in Groups 4 and 6, increase in CAT activity in Groups 5 and 6, and decrease in GSH-Px activity in Groups 4 and 6. Furthermore, in comparison to the control group, significant differences were observed with respect to certain serum biochemical parameters, including decrease in glucose levels in Groups 5 and 6, decrease in triglyceride levels in Groups 2 and 4, decrease in cholesterol levels in Groups 2 and 5, decrease in the total protein level of Groups 4−6, decrease in the ALT activity of Groups 5 and 6, increase in the AST activity of Group 4, decrease in the ALP activity of Groups 2−6 and increase in the uric acid level of Group 2. In the groups that were administered propolis in association with fluoride, improvement was observed in some oxidative stress parameters and certain other biochemical parameters. Changes determined in the oxidative stress parameters (especially MDA and SOD) were indicative of the anti-radical activity of propolis on the free radicals generated by sodium fluoride. However, the values not drawing completely close to those of the control group can be explained with propolis not being able to completely eliminate the free radicals and the other adverse effects generated by fluoride.     

Fenthion induced-oxidative stress in the liver of adult rats and their progeny: Alleviation by Artemisia campestris

Fenthion (FEN) is an organophosphate insecticide used in both agricultural and urban areas throughout the world including Tunisia. Recent investigations have proved the crucial role of natural antioxidants to prevent the damage caused by toxic compounds. In this study, we investigated the role of Artemisia campestris (Ac) leaf powder in protection against oxidative damage and hepatotoxicity induced by fenthion in female rats and their pups. Female Wistar rats were divided into four groups: group I served as controls which received standard diet, group II received orally FEN 551 ppm, group III received both 551 ppm of FEN and experimental diet (5% Artemisia) and group IV received experimental diet (5% Artemisia). Oral administration 551 ppm of FEN by drinking water to adult rats caused hepatotoxicity as monitored by the increase in the levels of hepatic markers enzymes (transaminases and lactate dehydrogenase), total cholesterol (TC) and triglycerides (TG), as well as hepatic malondialdehyde (MDA) levels thus causing a drastic alteration in antioxidant defence system. Particularly, the activities of catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) and the level of reduced glutathione (GSH) increased by FEN. These biochemical alterations were accompanied by histological changes marked by leucocytes infiltration, sinusoidal dilatation (moderate peliosis), granuloma inflammatory disorders and necrosis in hepatocytes of dams. While, slight leucocytes infiltration was shown in pups. Treatment with Ac prevented the liver damage induced by FEN, as revealed by inhibition of hepatic lipid peroxidation accompanied by an improvement of liver histopathological changes, CAT and GPx activities except GSH and SOD which were not modified. It could be concluded that A. campestris is promising a protective agent against hepatotoxicity during the exposure to fenthion.     

Agrobacterium induces plant cell death in wheat (Triticum aestivum L.)

The symptoms of gall or hairy root do not occur in the interactions between wheat (Triticum aestivum L.) and other monocotyledonous plants, with Agrobacterium tumefaciens or Agrobacterium rhizogenes. However, both bacteria colonized wheat root surfaces at similar levels (2.0 × 10⁷ colony forming U g⁻¹ root) and grew without inhibition in suspension with intact or wounded wheat embryos or root segments present. Suspension-cultured wheat embryo cells grown in 7.4 m M O₂ displayed 23% cell death after 1 h exposure to Agrobacterium cells, while the extent of cell death with 2.1 m M O₂ averaged 8%. Cultured wheat embryo and root cells rapidly produced hydrogen peroxide (H₂O₂) when contacted with A. tumefaciens or A. rhizogenes. Production of H₂O₂ was lower at 2.1 m M O₂ than 7.4 mM O₂. Browning and autofluorescence of epidermal cells of callus derived from wheat embryos and wheat roots was observed after inoculation with Agrobacterium. An increase in ferulic acid was detected in the walls of roots exposed to Agrobacterium. However, neither lignin nor callose was detected by diagnostic staining methods. These findings suggest that Agrobacterium induced a resistance-like response in wheat that may reduce the efficacy of transformation and limit the normal symptom formation.     

Biochemical,physiological and morfological responses in common carp (Cyprinus carpio L.) after long-term exposure to terbutryn in real environmental concentration

The effects of terbutryn at concentrations of 0.02 (reported concentration in Czech rivers), 0.2, and 2.0 μg l^?1 were assessed in one-year-old common carp (Cyprinus carpio L.) exposed for 90 days. Influence on biometric parameters, hematology, biochemistry, histology, and oxidative stress was investigated. Exposure to 0.02, 0.2 and 2.0 μg l^?1 showed significant differences oxidative stress biomarkers compared to controls but exposure to 0.2 and 2.0 μg l^?1 significantly affected biochemical and hematological profiles. Long-term exposure of terbutryn in carp resulted in slight alterations in internal organs and increased reactive oxygen species formation, resulting in oxidative damage to lipids and proteins and inhibition of antioxidant capacities.     

Metabolism and toxicity of metribuzin in mouse liver

Metribuzin was hepatotoxic in mice when administered intraperitoneally (ip) at sublethal doses of 150 to 250 mg/kg. Four dose-dependent abnormalities were evident. Histopathological examination revealed a fulminant centrilobular hepatic necrosis. The serum glutamic-pyruvic transaminase (GPT) activity was elevated. The liver glutathione (GSH) content was almost completely depleted. There was extensive covalent binding of radiocarbon from [carbonyl-¹⁴C]metribuzin to liver proteins and also high blood levels of metribuzin fragments. Each of these four effects of metribuzin on the liver or blood was alleviated or blocked in mice pretreated with piperonyl butoxide (PB), which inhibits the cytochrome P-450-dependent monooxygenase. PB also reduced the lethality of metribuzin by three-fold. In contrast, pretreatment with diethyl maleate to suppress the liver GSH content increased the lethality of metribuzin by twofold. The hepatotoxicity and acute lethality of metribuzin were probably due to reactive intermediates which are normally detoxified by GSH conjugation. The principal urinary metabolites of metribuzin in mice and rats are mercapturic acids, which arise via metribuzin sulfoxide or deaminometribuzin sulfoxide reacting with GSH. Sulfoxidation therefore appears to activate metribuzin to an electrophilic metabolite which, in the absence of GSH, binds to tissue proteins producing hepatotoxicity.     

Role of sucrose in the development of Fusarium wilt in lupine embryo axes

The response of embryo axes of Lupinus luteus L. cv. Polo to invasion by Fusarium oxysporum Schlecht f.sp. lupini was studied 84, 96 and 108 h after inoculation and under various trophic conditions. Four variants were compared: these included inoculated embryo axes cultured with or without 60 mM sucrose (+Si and −Si) and non-inoculated embryo axes cultured with or without 60 mM sucrose (+Sn and −Sn). The exogenous sucrose caused an accumulation of sucrose, glucose and fructose in +Sn embryo axes. Pathogen invasion generally resulted in a sharp decline in soluble carbohydrate content of +Si embryo axes. Symptoms of Fusarium wilt included growth inhibition of the embryo axes and a decrease in their fresh weight, which was particularly strong in −Si axes. While the addition of exogenous sucrose to the medium 24 h prior to inoculation significantly alleviated the disease, its application 48 h after inoculation did not restrict disease development. In +Si embryo axes, the activity of peroxidases (EC 1.11.1.7), covalently and ionically bound to the cell walls, increased considerably, mainly up to 96 h after inoculation. This increase was significantly higher than that seen in +Sn embryo axes. These results are in contrast to those of an earlier study [Morkunas I, Marczak Ł, Stachowiak J, Stobiecki M. Sucrose-stimulated accumulation of isoflavonoids as a defense response of lupine to Fusarium oxysporum. Plant Physiol Biochem 2005;43:363–73], which showed a pronounced decrease in the flavonoid glycoside content of +Si tissues. Furthermore, starting 96 h after inoculation, a decrease in phenylalanine ammonialyase activity (PAL, EC 4.3.1.5) was observed in inoculated tissues, in comparison to non-inoculated tissues. The decrease was especially pronounced in +Si tissues. However, a significant increase in the free isoflavone aglycones content of +Si embryo exes, i.e. genistein, 2′-hydroxygenistein, wighteone and luteone. Simultaneously, a strong increase in the activity of β-glucosidase (EC 3.2.1.21), hydrolyzing isoflavone glycosides, was observed in inoculated tissues. The activity of these enzymes was much higher in −Si embryo axes than in +Si embryo axes. The results suggest that soluble carbohydrates may play a role in the restriction of Fusarium wilt development, as they may be related both to intensified lignification and to the antimicrobial role of free aglycones, which may be products of the hydrolysis of isoflavone glycosides generated prior to the invasion stage.     

Toxic effects of acephate on Paramecium caudatum with special emphasis on morphology,behaviour, and generation time

The continuous increase in the number of new chemicals as well as the discharges of solid and liquid wastes triggered the need for simple and inexpensive bioassays for routine testing. In recent years, there has been increasing development of methods (particularly rapid tests) for testing environmental samples. This paper describes the quick toxic evaluation of an organophosphorus insecticide, acephate (O,S-dimethyl acetylphosphoramidothioate) on Paramecium caudatum for acute and sub-acute toxicity studies with reference to morphology, behaviour, and its generation time. The lethal concentrations for 10 min and 2 h were determined by probit method, as 500 mg L⁻¹ and 300 mg L⁻¹, respectively. Higher concentrations of 10 min exposure caused cell lysis with disintegration of cell membrane and precipitation of protoplasm. Combination of conventional light microscopy and computerized video tracking systems were used to study the locomotor behaviour of paramecia. The test organism was under stress and exhibited an initial increase and subsequent decrease in the swimming speed when exposed to 1/4, 1/2, 3/4, and LC₅₀ concentrations for 10 min (125, 250, 375, and 500 mg L⁻¹, respectively). Similar changes were also noticed when paramecia were exposed to LC₅₀ for 2 h. In a separate set of experiments, the number of generations and generation time in 24 h was evaluated with respect to the different sub-lethal concentrations (30, 60, 120, and 240 mg L⁻¹). The number of generations decreased and generation time extended significantly in a concentration dependent manner. The results indicate that the Paramecium toxicity assay could be used as a complimentary system to rapidly elucidate the cytotoxic potential of insecticides. The major advantages associated with these tests are: they are inexpensive, simple, user-friendly, space saving, and seem to be attractive alternatives to conventional bioassays.