Endocrine disruption and metabolic changes following exposure of Cyprinus carpio to diethyl phthalate

Diethyl phthalate (DEP) enter into aquatic environment from industries manufacturing cosmetics, plastic and many commercial products and can pose potential fish and human health hazard. This experiment evaluated effects of DEP in adult male (89 g) common carp (Cyprinus carpio) by exposing them to fractions of LC₅₀ (1/500-1/2.5) doses with every change of water for 28 days. Vitellogenin induction metabolic enzymes, somatic indices and bioaccumulation were studied on 7th, 14th, 21st and 28th day. The 96th hour LC₅₀ of DEP in fingerlings was found to be 48 mg/L. Compared to control, except increase (P < 0.01) in alkaline phosphatase activity (EC 3.1.3.1) and liver size, there was decrease (P < 0.01) in activity of acid phosphatase (EC 3.1.3.2), aspartate aminotransferase (EC 2.6.1.1), alanine aminotransferase (EC 2.6.1.2) and testiculosomatic index following exposure to 1, 5 and 20 ppm DEP. Significant (P < 0.01) dose dependant vitellogenin induction was observed with exposure of fish to 0.1, 1 and 5 ppm DEP. The bioaccumulation of DEP in testis, liver, brain, gills and more importantly in muscle tissues of fish increased significantly (P < 0.01) with increase of dose from 1 to 5 ppm. Significant interaction (P < 0.01) of dose and duration of exposure indicated that exposure period of a week to two was sufficient to bring about changes in quantifiable parameters studied. Fish exposed to 20 ppm DEP became lethargic and discolored during onset of the 4th week. This is the first report describing metabolic changes and vitellogenin induction following exposure of C. carpio to DEP dose that is as low as 1/500th fraction of LC₅₀.     

Acute toxicity of organophosphorous pesticide diazinon and its effects on behavior and some hematological parameters of fingerling European catfish (Silurus glanis L.)

Diazinon is commonly used for pest control in the agricultural fields surrounding freshwater reservoirs. So this study was conducted to determine the acute toxicity of this organophosphorous pesticide, contaminating aquatic ecosystems as a pollutant, and its effects on behavior, and some hematological parameters of fingerling European catfish, Silurus glanis. Diazinon was applied at concentrations of 1, 2, 4, 8, 16, 32, and 64 mg L⁻¹. The water temperature in the experimental units was kept at 16 ± 1 °C. The number of dead fishes significantly increased in response to diazinon concentrations 2-64 mg L⁻¹ (p < 0.05). With increasing diazinon concentrations, the fishes exposed duration 1 to 96 h significantly increased the number of dead fishes (p < 0.05 for each cases). The 1, 24, 48, 72, and 96 h LC₅₀ values (with 95% confidence limits) of diazinon for fingerling European catfish were estimated as 14.597 (12.985-16.340), 12.487 (11.079-14.471), 8.932 (7.907-10.348), 6.326 (no data because of p > 0.05), and 4.142 (no data because of p > 0.05) mg L⁻¹, respectively. Compared to the control specimens, fish after an acute exposure to diazinon was significantly lower erythrocyte, leukocyte, hemoglobin, hematocrit, MCV, MCH, and MCHC values (p < 0.05). In addition, it was also showed a significantly negative correlation between these hematological parameters and exposure times of diazinon (p < 0.01).     

Effect of fenvalerate on oxidative stress biomarkers in the brackish water prawn Penaeus monodon

The toxicity of fenvalerate to the prawn Penaeus monodon was evaluated using biomarkers of stress. In a preliminary bioassay test, P. monodon was exposed to a series of fenvalerate concentrations, which showed 4, 6.5 and 8.5 μg L⁻¹ to be sublethal, median lethal and lethal, respectively. Sublethal effect of fenvalerate was further evaluated in hepatopancreas, muscle and gills of prawns with reference to oxidative stress biomarkers. Significant induction of lipid peroxidation and glutathione-S-transferase activity was found in hepatopancreas, muscle and gills of prawns exposed to fenvalerate when compared to control (P < 0.001, P < 0.05 and P < 0.05). On the contrary, the activities of Superoxide dismutase, catalase, glutathione peroxidase, vitamin C, vitamin E and glutathione were found to be reduced in the experimental group of prawns when compared to control. The results suggest that the animals were under oxidative stress when exposed to sublethal concentration of fenvalerate.     

Biological and biochemical parameters of Biomphalaria alexandrina snails exposed to the plants Datura stramonium and Sesbania sesban as water suspensions of their dry powder

Four plant species, as a dry powder of their leaves, were tested against Biomphalaria alexandrina snails, the intermediate host of Schistosoma mansoni. The bioassay tests revealed the plants Datura stramonium and Sesbania sesban to be more toxic to the snails than the other two ones. Therefore, they were tested against snails’ fecundity (M_x), reproduction rate (R_o) and their infection with S. mansoni miracidia. In addition, total protein concentration and the activities of the transaminases (AsT and AlT) and phosphatases (AcP and AkP) enzymes in hemolymph and tissues of snails treated with these plants were determined. As well, glucose concentration in snails’ hemolymph was evaluated.Exposure of snails for 4 weeks to LC₁₀ and LC₂₅ of the plants D. stramonium and S. sesban dry powder markly suppressed their M_x and R_o. The reduction rates of R_o for snails exposed to LC₂₅ of these plants were 62.1% and 76.4%, respectively. As well, a considerable reduction in the infection rates of snails exposed to these plants either during, pre- or post-miracidial exposure was recorded. Thus, infection rates of snails treated during miracidial exposure with LC₁₀ of D. stramonium and S. sesban were 41.7% and 52.2%, respectively, compared to 92.6% for control group (P < 0.01). These plants, also, reduced the duration of cercarial shedding and cercarial production/snail. So, snails exposed to LC₂₅ of these plants shed 372.8 and 223.2 cercariae/snail, respectively, compared to 766.3 cercariae/infected control snail (P < 0.01).The results, also, revealed that glucose and total protein concentrations in hemolymph of snails treated with LC₁₀ and LC₂₅ of these plants were decreased, meanwhile, the activities of the enzymes AsT, AlT, AcP and AkP were elevated (P < 0.01). However, the activity of AcP in tissues of treated snails was decreased compared to that of control ones. It is concluded that LC₂₅ of the plants D. stramonium and S. sesban negatively interferes with biological and physiological activities of B. alexandrina snails, consequently it could be effective in interrupting and minimizing the transmission of S. mansoni.     

Fenvalerate induced stress mitigation by dietary supplementation of multispecies probiotic mixture in a tropical freshwater fish, Labeo rohita (Hamilton)

Five experimental diets with various combinations of probiotics, namely T1 & T6 (basal feed (BF) without probiotics), T2 & T7 (BF + Bacillus subtilis + Lactococcus lactis), T3 & T8 (BF + L. lactis + Saccharomyces cerevisiae), T4 & T9 (BF + B. subtilis + S. cerevisiae) and T5 & T10 (BF + B. subtilis + L. lactis + S. cerevisiae) were fed to Labeo rohita fingerlings for 30 days. Treatment groups T1, T2, T3, T4 and T5 were exposed to Fenvalerate, at a concentration of 1.79 μg L⁻¹. The SOD and CAT activity was significantly affected (P < 0.01) in fenvalerate treated groups. However, the supplementation of the three-probiotic mixture at equal concentration showed markedly reduced activity. Similarly, RBC, Hb, NBT, total protein and albumin values were reduced significantly (P < 0.01) in the fenvalerate exposed fish as compared to the probiotic supplemented fish. Fenvalerate exposure also showed increased serum ALP, ACP and Bilirubin values (P < 0.01) in comparison to the non-exposed fish. Histological observations of the gills, kidney and liver showed tissue degeneration after fenvalerate exposure, which however showed marked recovery on the three-probiotic mixture supplementation. Therefore, these results indicate that a mixture of multi-species probiotic supplementation in equal concentration acts beneficially in mitigating the stressful effects of fenvalerate.     

Anti-oxidative and immuno-hematological status of Tilapia (Oreochromis mossambicus) during acute toxicity test of endosulfan

Endosulfan is an organochlorine pesticide widely used in agriculture and hence finds its way into natural water bodies, thus affecting aquatic life. The purpose of this study was to determine LC₅₀ of endosulfan (99%; α:β ratio of 7:3) in Tilapia, Oreochromis mossambicus and study its effect on anti-oxidative enzymes (superoxide dismutase, glutathione-S-transferase and catalase), immuno-hematological profile (RBC, WBC, Hb, serum protein, albumin-A, globulin-G, A/G ratio, phygocytic activity as indicated by nitroblue tetrazolium reduction, serum cortisol and serum lipid peroxidation) and neurotransmitter acetylcholine esterase enzyme activity. The LC₅₀ value at 96 h and 95% confidence limit for tilapia (46.78 g) was estimated as 3.6 μg/L. Activities of anti-oxidative enzymes, immuno-hematological profile, blood glucose and neurotransmitter activity was significantly influenced (P < 0.01) in dose dependent manner. This was reflected in the behavior of fish that was altered from normal during acute toxicity.     

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.     

Acute toxicity and histopathological effects of sublethal fenitrothion on Nile tilapia, Oreochromis niloticus

Organophosphothionate insecticide fenitrothion is known as potential toxic pollutant contaminating aquatic ecosystems. The effects of fenitrothion were studied to determine the 96 h LC₅₀ value on Nile tilapia (Oreochromis niloticus) and investigate histopathological responses of fish exposed to sublethal fenitrothion concentrations. Data obtained from the fenitrothion acute toxicity tests were evaluated using the Probit Analysis Statistical Method. The 96 h LC₅₀ value and 95% confidence limit for Nile tilapia (58.70 ± 6.97 g) was estimated as 0.84 (0.68-1.15) mg/L. Behavioral changes were observed closely during the acute toxicity test. The bioassay experiments were repeated three times and static test method was used. Some fish exposed to 96 h 0.1, 0.5 mg/L fenitrothion concentrations showed histopathological alterations in the gills, liver, kidney, brain and testes. Severely deformations were observed at 0.5 mg/L fenitrothion on the gills lamella such as hyperemia, epithelial hyperplasia, fusion and telangiectasis, in the liver tissue such as cloudy swelling, hydropic degenerations and lipid infiltration. In addition hyperemia and hemorrhage observed in kidney tissue and hyperemia was determined in brain tissue.     

One-tenth dose of LC50 of 4-tert-butylphenol causes endocrine disruption and metabolic changes in Cyprinus carpio

Of the huge annual worldwide production (500,000 MT in 1997) of alkylphenol polyethoxylates (APEs) that are widely used as nonionic surfactants and anti-oxidants in variety of products, 60% ends up in water bodies. They undergo biodegradation to form octyl-, butyl-, and nonyl-phenols. This experiment evaluated effects of 4-tert-butyl phenol (4-TBP) in Cyprinus carpio, a projected candidate species in sewage fed fisheries. The 96th h LC₅₀ of 4-TBP was found to be 6.9 mg/L. Fishes were treated with 1/10th (0.69 mg/L), 1/5th (1.38 mg/L), and 1/3rd (2.3 mg/L) dose of LC₅₀. Whereas there was significant (P < 0.01) decrease in alkaline phosphatase [EC 3.1.3.1] and aspartate aminotransferase [EC 2.6.1.1] activity; alanine aminotranferase [EC 2.6.1.2] and acid phosphatase [3.1.3.2] (except decrease at 1/10th dose of LC₅₀) activity, vitellogenin production in muscle and hepatic- and reno-somatic indices were increased compared to control. With all the dose levels tested, testicular-somatic index (testis size) was reduced (P < 0.01) and histo-architectural changes in testicular and liver tissue were found even in group given 1/3rd dose of LC₅₀.     

Effects of the organophosphate pesticide Folidol 600® on the freshwater fish, Nile Tilapia (Oreochromis niloticus)

Nile Tilapia (Oreochromis niloticus) juveniles were exposed to different concentrations of Folidol 600® in static toxicity tests. The 24, 48, 72 and 96 h LC₅₀ values of Folidol 600® to O. niloticus were 17.82, 8.91, 4.00 and 2.70 mg L⁻¹, respectively. The values of hematological parameters increased, and inhibition of cholinesterases activity (AChE, BChE and PChE) in plasma of fish exposed to the higher concentrations of pesticide reached 94%. Furthermore, the exposure of Tilapia to Folidol 600® caused an increase of 4%, 20% and 38.4% in oxygen consumption at 0.1, 0.5 and 1.0 mg L⁻¹, respectively. However, exposure to 2.5, 5.0 and 10 mg L⁻¹ caused a decrease of 33.6%, 35.2% and 42.4% in oxygen consumption relative to the control. The ammonium excretion of fish exposed to 0.0, 0.1, 0.5, 1.0, 2.5, 5.0 and 10.0 mg Folidol 600®/L was 0.12, 0.18, 0.30, 0.33, 0.37, 0.36 and 0.33 μg/g/min, i.e., 50%, 150%, 175%, 208%, 200% and 175% increase, respectively, relative to the control.     

Investigation of acute toxicity of (2,4-dichlorophenoxy)acetic acid (2,4-D) herbicide on crayfish (Astacus leptodactylus Esch. 1823)

The acute 96 h LC₅₀ of (2,4-dichlorophenoxy)acetic acid (2,4-D), a widely used agricultural herbicide, was determined on crayfish (Astacus leptodactylus Esch. 1823). Crayfish of 23.5 ± 1.49 g mean weight and 9.6 ± 0.21 cm mean length were selected for the bioassay experiments. The experiments were repeated three times, in 10 L tap water. The data obtained were statistically evaluated by the use of the E.P.A computer program based on Finney’s probit analysis method and the 96 h LC₅₀ value for crayfish was calculated to be 32.6 mg/L in a static bioassay test system. 95% lower and upper confidence limits for the LC₅₀ were 15.10-327.16. In conclusion, 2,4-D is highly toxic to crayfish, a non-target organism in the ecosystem. Water temperature was 23 ± 1 °C. Behavioral changes of crayfish were recorded for all herbicide concentrations.     

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.     

Ultra-structural changes in the integument induced by the use of three of six forms of allylisothiocyanate tested against Plutella xylostella and Pieris rapae larvae

The insecticidal activity of four forms of Hong Jing (HJ) allylisothiocyanate (AITC), AITC + cypermethrin (HJ_A, HJ_B, and HJ_C) with ratio of (1:1, 4:1, and 2:1), pure AITC (HJ_D), and two forms of Hong Du (HD) AITC, AITC + chlorpyrifos (HD_A and HD_B) with ratio of (2:1 and 2:1), respectively, were studied on the major cruciferous insect larvae Plutella xylostella (L.) and Pieris rapae (L.) by combining both spraying and dipping methods. The P. rapae was more susceptible than P. xylostella larvae. The LC₅₀ values 72 h after treatment of AITC forms (HJ_B, HJ_A, HJ_C, HJ_D, HD_B, and HD_A) on the P. rapae were; 0.07, 0.08, 0.16, 0.83, 0.26, 1.08 gL⁻¹, and 0.69, 0.26, 5.45, 0.93, 3.01, 5.98 gL⁻¹ on the P. xylostella, respectively. The toxicity of some of the AITC forms was very close to or better than that of the commercial contact insecticides such as chlorpyrifos (LC₅₀ = 0.03 and 0.04 gL⁻¹ on P. rapae and P. xylostella, respectively), and cypermethrin (0.65 and 0.78 gL⁻¹, respectively, against P. rapae and P. xylostella). The ultrastructural studies on the integument of the third larval instar of P. xylostella treated by sub-lethal concentration (LC₂₀) of HJ_B, HJ_D, and HD_B were carried out by using transmission electron microscope. The more pronounced alterations in the hypodermis and mitochondria cells. They exhibited changes in all treated samples. The hypodermis was almost completely destroyed, and the mitochondria exhibited morphological alterations, represented by enlargement, matrix rarefaction and vacuolization of the mitochondria matrix, quantity of cristae reduced, and density electron matrix lessened. These AITC forms have potential as contact insecticides, and the ultra structural observations confirm the insecticidal efficiency of different AITC forms on P. rapae and P. xylostella.     

Persistent sub-lethal chlorine exposure elicits the temperature induced stress responses in Cyprinus carpio early fingerlings

Thermal effluents discharged through cooling systems of nuclear power plants often contain chlorine (used to control bio-fouling), which may affect the metabolic status of fishes. In order to evaluate the hypothesis, we tested the effect of high temperature and a persistent sub-lethal chlorine exposure on stress responses in Cyprinus carpio advanced fingerlings. Fishes were acclimated to four different temperatures (26, 31, 33, and 36 °C) and maintained for 30 days in two different groups. Subsequently, one of the groups was exposed to persistent chlorine (0.1 mg L₋₁) for another 28 days and was compared with their respective temperature controls (without chlorine exposure). Sub-lethal doses of pollutants and increasing temperatures with in the tolerance range may not always register any morphological changes Therefore, we studied organ specific biochemical pathways viz. aspartate amino transferase, alanine amino transferase (enzymes of protein metabolism) in liver and muscle; fructose 1,6 diphosphatase (gluconeogenic pathway), in liver; pyruvate kinase, malate dehydrogenase, and lactate dehydrogenase (glycolytic pathway) in muscle; glucose-6-phosphate dehydrogenase (pentose phosphate pathway) in liver; alkaline phosphatase (phosphorus metabolism) in intestine, liver, and muscle; acetylcholine esterase (neurotransmitting enzyme) in brain, and adenosine triphosphate (for membrane transport) in gills at two different acclimation periods (14 and 28 days). The results indicate that C. carpio fingerlings demonstrated metabolic readjustments with increasing temperatures, in order to cope with energy demand of the cell. However, exposure to chlorine at higher temperatures affected protein metabolism, gluconeogenic pathway and subsequently glycolytic pathway, leading to an energy-limited condition. In addition, alteration of membrane transport and neurotransmission might be an early indication of cellular damage. Overall results indicate that persistent sub-lethal chlorine exposure elicits temperature induced stress response in C. carpio early fingerlings.     

The toxic effects of pyrethroid deltamethrin on the common carp (Cyprinus carpio L.) embryos and larvae

Deltamethrin, a synthetic pyrethroid pesticide contaminating aquatic ecosystems as a pollutant, was investigated in the present study for toxic effects on embryos and larvae of common carp, Cyprinus carpio as a model. The control and five test experiments were repeated five times. The water temperature in the experimental units was kept at 24 ± 1 °C. The number of dead embryos significantly increased in response to deltamethrin concentrations 0.005, 0.05, 0.5, 5, 25, and 50 μg L⁻¹ (p<0.05 for each cases). Dose-response decreases in hatching success were recorded as 75.2, 64.6, 47.4, 26.0, 14.4, and 9.0%, respectively. The lowest concentration of deltamethrin (0.005 μg L⁻¹) produced a significantly decrease in number of dead larvae compared to control group (p<0.05). With increasing deltamethrin concentrations, the larvae exposed duration 1-48 h significantly increased the number of dead larvae (p<0.05 for each cases). The 48 h LC₅₀ values (with 95% confidence limits) of deltamethrin for common carp embryos and larvae were estimated as 0.213 (0.103-0.404) and 0.074 (0.011-0.260) μg L⁻¹, respectively. The results provide evidence that deltamethrin pollution may have an adverse effect on the reproduction and development of carp, which should be considered when this chemical is used in agricultural areas near aquatic ecosystems.     

Physiological effect of chitinase purified from Bacillus subtilis against the tobacco cutworm Spodoptera litura Fab.

An extracellular chitinase was purified from Bacillus subtilis. The lethal concentration (LC₅₀) was determined by using chitinase in first, second, and third instars of Spodoptera litura Fab. Chitinase showed the highest insecticidal activity at 6 μM concentration within 48 h. The nutritional indices were also significantly affected by the 6 μM concentration (P < 0.05). Food consumption, efficiency of conversion of ingested and digested food, relative growth rate, and consumption values declined significantly while approximate digestibility was increased. Our study indicates that treatment of host plant leaves with the chitinase can regulate (reduce) larval growth and weight, and enhance the mortality. This may serve as an effective biocide and alternative to Bt toxin.     

Inhibition of Na-K-ATPase in different tissues of freshwater fish Channa punctatus (Bloch) exposed to monocrotophos

Freshwater fish, Channa punctatus, commonly known as the snakehead fish, was exposed to two sublethal concentrations (0.96 and 1.86 mg/L) (selected on the basis of 1/20 and 1/10 of 96 h LC₅₀ value) of monocrotophos for two exposure periods (15 and 60 days). Effects of monocrotophos on Na⁺, K⁺-ATPase in liver, kidney, muscle, intestine, brain, heart and gills were determined. Results indicate that Na⁺, K⁺-ATPase activity in tissues decreased as concentration of monocrotophos and exposure period increased. Monocrotophos induced significant inhibitory effects on the Na⁺, K⁺-ATPase activity of C. punctatus, ranging from gills (70%) > Kidney (63%) > brain (57%) > intestine (52%) > liver (50%) > muscle (47%) > heart (44%) inhibition at a sublethal concentration of 0.96 mg/L. Significant inhibition was detected in Na⁺, K⁺-ATPase activity, ranging from gills (90%) > heart (78%) > kidney (78%) > muscle (74%) > intestine (71%) > brain (67%) > liver (63%) at sublethal concentration of 1.86 mg/L. After subacute exposure (15 days) only gills and brain showed significant inhibition after higher concentration (1.86 mg/L). However, it is evident that exposure duration is more important than dose in the inhibition of the activity of enzyme. At lower concentration initial stimulation of the activity of Na⁺, K⁺-ATPase activity was also noticed. It is suggested that the inhibition of the ATPase by monocrotophos blocked the active transport system of the gill epithelial as well as chloride cells, glomerular and epithelial cells of the tubules and thus altered the osmoregulatory mechanism of the fish. In fact, the impairment of the activity of enzymes which carry out key physiological roles could cause alterations of the physiology of the whole organism.     

Effects of dietary pyridoxine on growth and biochemical responses of Labeo rohita fingerlings exposed to endosulfan

A sixty-day experiment was carried out to study the effect of dietary pyridoxine (PN) on growth performance, RNA/DNA ratio and some biochemical parameters of Labeo rohita fingerlings exposed to sub-lethal dose of endosulfan (1/10th of 96 h static non-renewal LC₅₀ = 0.2 ppb) to assess the role of pyridoxine in ameliorating the negative effects of endosulfan. Two hundred seventy fingerlings (6.5 ± 0.26 g) were randomly distributed into six treatments in triplicates (15 fish/tank). Five iso-nitrogenous (35.45-35.75% crude protein) purified diets were prepared with graded levels of pyridoxine. Six treatment groups were T₀ (10 mg PN + without endosulfan), T₁ (0 mg PN + endosulfan), T₂ (10 mg PN + endosulfan), T₃ (50 mg PN + endosulfan), T₄ (100 mg PN + endosulfan) and T₅ (200 mg PN + endosulfan). Weight gain (%), specific growth rate (SGR), tissue glycogen, and protease activity were significantly (P < 0.05) higher in pyridoxine fed groups compared to their non-pyridoxine fed counterpart. Protease activity was positively correlated (R² = 0.931) with (%) weight gain. Glucose-6-phosphate dehydrogenase (G6PDH) activity was significantly (P < 0.05) higher in non-pyridoxine fed group and decreased in pyridoxine fed counterparts. There were no significant (P > 0.05) effect of dietary pyridoxine on feed conversion ratio (FCR), protein efficiency ratio (PER), survival, gastro-somatic index (GSI), hepato-somatic index (HSI) and liver and muscle DNA levels of L. rohita fingerlings. RNA levels, both in liver and muscle, increased significantly (P < 0.05) in pyridoxine fed groups. A positive correlation was observed between growth and RNA levels, both in liver (R² = 0.91) and muscle (R² = 0.88). RNA/DNA ratio showed a third order polynomial relationship with dietary pyridoxine, both in liver (Y = −0.014x³ + 0.1613x² − 0.5333x + 0.7933, R² = 0.987) and muscle (Y = −0.0407x³ + 0.4763x² − 1.6358x + 2.4667, R² = 0.9345). The overall results obtained in present study indicated that dietary pyridoxine supplementation at 100 or 200 mg PN/kg diet ameliorates the negative effects of endosulfan and restores optimal growth of L. rohita fingerlings.     

Effect of Basudin, Selecron and the phytoalkaloid Colchicine (pesticides) on biological and molecular parameters of Biomphalaria alexandrina snails

The results showed that survival rates of Biomphalaria alexandrina snails, reproductive potential and hatchability of eggs were evaluated post exposure to Basudin, Selecron and Colchicine. As well, DNA and RNA changes in the cells of ovotestis-digestive gland complex of treated snails were estimated. The current molluscicide Bayluscide was used as a reference compound.The pesticide Selecron proved to be more toxic to B. alexandrina snails than Basudin and Colchicine. Juvenile snails were dead post 3 weeks of exposure to the sublethal concentration LC₀ of either Selecron or Basudin, while 26.75% of snails still alive at Bayluscide treatment. In addition, exposure of adult snails to LC₀ of Selecron for 24 h/week for 4 weeks markedly reduced their reproductive rate (R₀) by 89.9%. Moreover, snails’ eggs failed to hatch post 24 h of exposure to LC₉₀ of either Selecron, Basudin or Bayluscide. Electrophoretic analysis indicated a decrease in the molecular weight of intact DNA in the ovotestis-digestive gland complex of snails treated with 250 ppm of Colchicine and LC₂₅ of Selecron, as it scored 1.2 and 76 bp, respectively, compared to 166.46 bp for control group, while the vice versa was recorded for RNA intensity. It was concluded that the tested pesticides have deleterious effects on snails’ reproductive rate, their eggs and the intensities of DNA and RNA in their ovotestis-digestive gland complex. Therefore, it is expected that reaching of such pesticides to snails’ habitats in water courses during plant pests control could minimize the population density of the snails intermediate hosts of schistosomiasis, hence probably interrupt and reduce the transmission of this parasite.     

The effects of Artemisia annua L. and Achillea millefolium L. crude leaf extracts on the toxicity, development, feeding efficiency and chemical activities of small cabbage Pieris rapae L. (Lepidoptera: Pieridae)

The biological effects of two important medicinal plants, Artemisia annua L. and Achillea millefolium (L.) (viz, mortality, growth, and feeding indices as well as enzyme and non-enzymatic activities) were studied on small white Pieris rapae L a deleterious pest of cruciferous plants under controlled conditions (16:8 h L:D at 25 ± 1 °C and 65 ± 5% RH). The LC₅₀ and LC₂₅ values were 9.387% and 3.645% for A. annua L. and 4.19% and 1.69% for A. millefolium (L.), respectively. At the lowest concentration (0.625%), the deterrency was 29.826% and 44.185% for A. annua L. and A. millefolium (L.), respectively. Feeding indices were variously affected with changes in a number of parameters and an increase in larval and pupal duration. The activity level of alkaline phosphatase increased sharply while alanin and aspartate aminotransferases showed a sharp decrease. For non-enzymatic compounds, the amount of glucose and uric acid increased, but total protein and cholesterol decreased. These results indicate that these two medicinal plants might possess potential secondary metabolites that may be useful for controlling potential insect pests.