Determination of genotoxicity of Fenaminosulf by Allium and Comet tests

Genotoxic effects of Fenaminosulf, fungicide and micro-biocide, were examined by using mitotic index (MI), mitotic phase, and Comet assay on the root meristem cells of Allium cepa. In the Allium root growth inhibition test, EC₅₀ value was firstly determined as 25 ppm and, 12.5 (0.5 × EC₅₀), 25 (EC₅₀) and 50 (2 × EC₅₀) ppm concentrations of Fenaminosulf were introduced to onion tuber roots. Distilled water was used as a negative control. All obtained data were subjected to statistical analyses by using SPSS 15.0 for Windows software. For comparison purposes, Duncan multiple range tests by using one-way analysis of variance (ANOVA) were employed and p < 0.05 was accepted as significant value. While MI (except in 24 h at 12.5 and 50 ppm) and prophase index increased, metaphase, anaphase and telophase indexes decreased in all concentrations compared to control at each exposure time. A significant increase in DNA damage was also observed at the concentration of 25 ppm in 24 h, 25 and 50 ppm in 96 h by Comet assay.     

Cytogenetic effects of commercially formulated atrazine on the somatic cells of Allium cepa and Vicia faba

In the present study cytogenetic effects of atrazine herbicide, were examined on the root meristem cells of Allium cepa and Vicia faba. Test concentrations were chosen by calculating EC₅₀ values of formulated atrazine against both the test systems which determined to be 30 mg l⁻¹ for A. cepa and 35 mg l⁻¹ for V. faba, respectively. For cytogenetic effects root meristem cells of A. cepa were exposed to 15, 30 or 60 mg l⁻¹ whereas V. faba to 17.5, 35 or 70 mg l⁻¹ for 4 or 24 h. Roots exposed for 4 or 24 h, after sampling, were left in water for 24 h recovery and sampled at 24 h post-exposure. A set of onion bulbs or seedlings of V. faba exposed to DMSO (0.3%) was run parallel for negative control. Treatment of atrazine significantly and dose-dependently inhibited the mitotic index (MI) and induced micronucleus formation (MN) chromosome aberrations (CA) and mitotic aberrations (MA) in both the test systems at 4 or 24 h. Root meristem cells examined at 24 h post-exposure also revealed significant (p < 0.001) frequencies of MN, CA or MA despite considerable decline. Chromosome breaks and fragments were found to be major CA whereas C-metaphase, chromosome bridges and laggards were prevalent MA. Results of our study, indicate that atrazine may produce genotoxic effects in plants. Further, both the plant bioassays found to be sensitive indicators for the genotoxicity assessment as the outcome of majority of in vivo/in vitro mammalian tests are comparable.     

Determination of genotoxic effects of chlorfenvinphos and fenbuconazole in Allium cepa root cells by mitotic activity, chromosome aberration, DNA content, and comet assay

Genotoxic effects of Chlorfenvinphos and fenbuconazole were examined by using mitotic index, mitotic phase, chromosomal abnormalities, 2C DNA content and Comet assay on the root meristem cells of Allium cepa. The roots were treated with 10, 20, 40, 60, 80, and 100 ppm concentrations for 24 and 48 h. The results indicated that Chlorfenvinphos and fenbuconazole significantly decreased the mitotic index in all treatments when compared with their controls. The percentages of mitotic phases have changed. Chlorfenvinphos and fenbuconazole significantly increased the abnormal cell frequency at all concentrations and treatment periods when compared with their controls. Different abnormal mitotic figures were observed in all mitotic phases. Among these abnormalities were stickiness, anaphase bridges, c-mitosis, laggards, and micronucleus. These pesticides remarkably depressed the 2C DNA content in the root meristems of A. cepa. The genotoxicity of chlorfenvinphos and fenbuconazole in A. cepa root cells was analyzed using comet assay, which allows the detection of single strand breaks. In all concentrations, chlorfenvinphos and fenbuconazole induced a significant increase in DNA damage. Additionally, it was also researched to determine if there is a relation between the amount of DNA and the DNA damage and a regression analyses was conducted. When the data that was accumulated via comet analysis from A. cepa root tip cells that are treated with type chlorfenvinphos and fenbuconazole, was compared to the data that was acquired as the result of the measurement of 2C DNA amount, a relation with negative correlation was found, (respectively, r = −0.80 and r = −0.82). This relation factor is statistically important and strong (p < 0.05).     

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.     

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.     

Food consumption, utilization, and detoxification enzyme activity of the rice leaffolder larvae after treatment with Dysoxylum triterpenes

The toxicity and physiological (enzyme and nutritional indices) effect of Dysoxylum triterpenes 3β,24,25-trihydroxycycloartane and beddomei lactone were evaluated on the rice leaffolder Cnaphalocrocis medinalis (Guenée). The LC₅₀ [6.66 ppm (SD = 0.31), 5.79 ppm (SD = 0.33) for 3β,24,25-DHCL and BL, respectively] and LC ₉₀ [14.63 ppm (SD = 0.36), 13.49 ppm (SD = 0.27) for 3β,24,25-DHCL and BL, respectively] were identified by probit analysis. Fourth instars were exposed to various concentrations (1.5, 3, 6, and 12 ppm) of Dysoxylum triterpenes. Results showed that treated larvae exhibited reduced food consumption and enzyme activity. Food consumption, digestion, relative consumption rate, efficiency of conversion of ingested food, efficiency of conversion of digested food, and relative growth rate values declined significantly but the approximate digestibility of treated larvae was significantly higher as a result of treatment (in particular 6 and 12 ppm). Likewise, the gut enzymes acid phosphatases, alkaline phosphatases, and adenosine triphosphatases were significantly inhibited by the Dysoxylum triterpenes. The high biological activity of these triterpenes from Dysoxylum sp. could be used as an active principle during the preparation of botanical insecticides for insect pest like rice leaffolder.     

Combined cytogenetic and ultrastructural effects of substituted urea herbicides and synthetic pyrethroid insecticide on the root meristem cells of Allium cepa

Combined cytogenetic and ultrastructural effects of substituted urea herbicides-isoproturon (ISO) or diuron (DIU) and a synthetic pyrethroid insecticide-deltamethrin (DEL) were examined in the root meristem cells of Allium cepa. For cytogenetic analysis root meristem cells were exposed to the mixtures of ISO (25 or 50 ppm) or DIU (20 or 40 ppm) and DEL (0.25 or 0.5 ppm) for 6 h and analyzed at 24 or 48 h post-exposure whereas for ultrastructural studies roots were exposed for 6 or 24 h to similar concentrations of combinations and examined. Both the combinations, ISO + DEL or DIU + DEL, were found to induce chromosomal breaks and variety of mitotic aberrations at 24 and 48 h post-exposure. The combinations containing higher concentration of DEL (0.5 ppm) induced statistically significant (p < 0.001) frequencies of aberrations than that of the combinations containing low concentration of DEL (0.25 ppm). Chromosome aberrations in all the treatment groups were less frequent than that of mitotic aberrations. Electron microscopic study revealed drastic alterations in the membranous organelles like concentric arrangement of endoplasmic reticulum, crescented or circular structure of Golgi complex dictyosomes and swollen mitochondria. Further, the combination of DIU + DEL appeared to be more toxic than that of ISO + DEL. Present findings suggest that the coexistence of ISO or DIU and DEL in plants synergies the toxicity inducing drastic ultrastructural alterations which are different from its independent effects reported earlier.     

Effects of different treatment methods of the fungicide jinggangmycin on reproduction and vitellogenin gene (Nlvg) expression in the brown planthopper Nilaparvata lugens Stål (Hemiptera:Delphacidae)

Jinggangmycin is an antibiotic fungicide against the rice sheath blight, Rhizoctonia solani, in China. Previous investigations have shown that jinggangmycin leaf spray stimulated the fecundity of Nilaparvata lugens Stål (Hemiptera:Delphacidae), but the effects of different application methods and concentrations of jinggangmycin on the reproduction of N. lugens have not been investigated. Here, we investigated three treatment methods (foliar and stem sprays and topical treatment) and four concentrations (100, 200, 400 and 800 ppm) of jinggangmycin on the reproduction of adult female N. lugens. The results showed that leaf spray significantly stimulated the fecundity of N. lugens and increased the vitellin content in female ovaries and the gene expression level of vitellogenin (Nlvg), but there was no significant effect on reproduction found for stem spray. In four concentrations of jinggangmycin, leaf sprays at concentrations of 100, 200 and 400 ppm and topical treatments of 100 and 200 ppm significantly increased the number of eggs laid, the vitellin content and the gene expression level of Nlvg compared to the control. Therefore, we suggest that stem spray at high concentration should be applied (if possible) when jinggangmycin is used against rice sheath blight to facilitate harmonious control of rice pests.     

Sensitivity of the mitotic cells of barley (Hordeum vulgare L.) to insecticides on various stages of cell cycle

In this study, the genotoxic effects of insecticides in different phases of cell cycle were investigated in the mitotic cells of barley (Hordeum vulgare L.). The seeds of H. vulgare L. Var. Karan 4 were treated with different concentrations (0.05%, 0.1% and 0.5%) of insecticides alphamethrin (AM) and monocrotophos (MP) for 6 h after synchronization of cells in G₁, S and G₂ phase of cell cycle with the help of various presoaking durations (7 h, 17 h and 27 h.). Positive and negative control was run parallel in the form of ethyl methane sulphonate (EMS) and distilled water, respectively. The data indicate that higher dose of alphamethrin and monocrotophos produce toxicity, chromosomal aberrations and mitotic aberrations in Hordeum vulgare L. The present study indicates the genotoxic potential of the insecticides AM and MP and it also showed that S-phase of cell cycle was more sensitive compared to the G₁ and G₂ phases.     

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.     

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).     

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.     

Ecophysiological category based toxicological responses in metabolism of earthworms: Impact of a pyrethroidal insecticide

Effects of alphamethrin (0.04 ppm, 0.08 ppm, 0.16 ppm, 0.32 ppm) on some metabolic dehydrogenases and proteins for 1, 2, 3, 4, 8 and 16 days using three ecologically different earthworm species (Perionyx sansibaricus, Lampito mauritii and Metaphire posthuma) were studied. The first significant effect was on 2nd/3rd day and maximum response was achieved on 16th day of exposure of alphamethrin at all concentrations. Similarly, maximum effect was obtained at 0.32 ppm alphamethrin at different exposure periods. It showed a dose- and duration-dependent inhibitory effects of a pyrethroid on enzymes and proteins of earthworms. There were approximately 47%, 43% and 41% declines in specific activities of cytoplasmic malate dehydrogenase (cMDH) of P. sansibaricus, L. mauritii and M. posthuma, respectively, in response to 0.32 ppm exposure of alphamethrin for 16 days. In case of similar treatment, specific activities of mitochondrial malate dehydrogenase (mMDH) showed 57%, 51% and 45% reductions in epigeic (P. sansibaricus), anecic (L. mauritii) and endogeic (M. posthuma) earthworms, respectively. Exposure of alphamethrin (0.32 ppm) also decreased the specific activities of lactate dehydrogenase (LDH) by 47% (P. sansibaricus), 35% (L. mauritii) and 39% (M. posthuma) in different types of earthworms. The reductions in specific activities of a Krebs cycle enzyme (mMDH) were greater than that of a glycolytic enzyme (LDH). The protein contents declined significantly in the earthworms exposed to alphamethrin. The decreases of 41%, 36% and 30% were obtained in cytoplasmic proteins of P. sansibaricus, L. mauritii and M. posthuma, respectively. However, the mitochondrial proteins showed reductions of 45% in P. sansibaricus, 46% in L. mauritii and 38% in M. posthuma in response to alphamethrin (0.32 ppm) intoxication for 16 days. The decrease in the protein content reflected an inhibitory effect of alphamethrin on protein turnover. The most pronounced effect of alphamethrin was on metabolic enzymes and proteins of P. sansibaricus and least effect was found on M. posthuma. This clearly showed that surface dwelling species of earthworms (epigeic) are more vulnerable to toxic chemical than deep burrowing species (endogeic). Thus a possible ecophysiological link exists between the toxicological responses and ecological categories at metabolic level in tropical earthworms.     

Effect of waterborne benomyl on the hematological and antioxidant parameters of the Nile tilapia, Oreochromis niloticus

The present study was conducted to determine the 96 h-LC₅₀ of benomyl to the Nile tilapia, Oreochromis niloticus and to investigate the biochemical or hematological indices of blood and the alterations in the antioxidant enzymes of this fish in response to sublethal concentrations of benomyl. Fish weighing 71.61 ± 12.05 g were used in this study; they were subjected to fasting for 4 weeks before treatment. An aqueous solution of benomyl (0, 0.5, 1, 2, 4, 8, and 16 mg L⁻¹) was administered for 96 h to determine the LC₅₀. The 96 h-LC₅₀ value of benomyl was 4.39 (3.23-5.60) mg L⁻¹ in the present study. For 5 weeks, the aqueous solution of benomyl (0, 100, 200, and 400 μg L⁻¹) was administered to investigate its effect on the hematological parameters and antioxidant enzymes. The predominant hematological findings in fish exposed to benomyl were as follows: no significant change in the Hb (g dL⁻¹) level, MCV (μm³), MCH (pg) and MCHC (%) as compared to the control. Benomyl exposure led to greater increases in the GPT, GOT (Karmen-unit), LDH (Wroblewski unit), total cholesterol, Fe, and Ca (mg dL⁻¹) values, whereas the levels of ALP (KA unit), total protein, triglyceride, albumin, and Mg (mg dL⁻¹) did not increase. Benomyl increased the in vivo HSI (%), GST (nmol min⁻¹ mg protein⁻¹), and SOD (U mg protein⁻¹) values in the fish livers in the test group, unlike those in the control group for 5 weeks. At concentrations higher than 100 μg L⁻¹, benomyl affected the GST and SOD levels of Nile tilapia in a dose- and time-dependent manner. The present findings suggest that the in vivo hepatotoxicity associated with benomyl may, in part, result from the hematological index, and antioxidants may provide limited protection against benomyl toxicity.     

Methylparathion- and carbofuran-induced mitochondrial dysfunction and oxidative stress in Helicoverpa armigera (Noctuidae: Lepidoptera)

The cotton bollworm, Helicoverpa armigera is a polyphagous pest of several crops in Asia, Africa, and the Mediterranean Europe. Organophosphate and carbamate insecticides are used on a large-scale to control Helicoverpa. Therefore, we studied the effect of methylparathion and carbofuran, an organophosphate and carbamate insecticide, respectively, on oxidative phosphorylation and oxidative stress in H. armigera larvae to gain an understanding of the different target sites of these insecticides. It was observed that state III and state IV respiration, respiratory control index (RCI), and P/O ratios were inhibited in a dose-dependent manner by methylparathion and carbofuran under in vitro and in vivo conditions. Methylparathion and carbofuran inhibited complex II by ∼45% and 30%, respectively. Lipid peroxidation, H₂O₂ content, and lactate dehydrogenase (LDH) activity increased and glutathione reductase (GR) activity decreased in a time- and dose-dependent manner in insecticide-fed larvae. However, catalase activity was not affected in insecticide-fed larvae. Larval growth decreased by ∼64% and 67% in larvae fed on diets with 100 μM of methylparathion and carbofuran. The results suggested that both the insecticides impede the mitochondrial respiratory functions and induced lipid peroxidation, H₂O₂, and LDH leak, leading to oxidative stress in cells, which contribute to deleterious effects of these insecticides on the growth of H. armigera larvae, along with their neurotoxic effects.     

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.     

Biological and biochemical responses of Biomphalaria alexandrina to some extracts of the plants Solanum siniacum and Artemisia judaica L.

The molluscicidal activity of cold water, boiled water, methanol, ethanol, acetone and chloroform extracts of Solanum siniacum and Artemisia judaica L. plants against Biomphalaria alexandrina snails was carried out. The tests revealed plant’s ethanol extract was more toxic to the snails than the other tested extracts. Therefore, it was tested against snails’ fecundity (M_x), reproduction rate (R₀) and their infection with Schistosoma mansoni miracidia. In addition, biochemical parameters and the activities of some enzymes in tissues of snails treated with the two tested plants were determined. As well, glucose concentration in snails’ hemolymph was evaluated. Exposure of B. alexandrina snails to plant’s ethanol extract led to a significant reduction in their survival and snails’ fecundity, reproduction rate. In addition, it caused a considerable reduction in the infectivity of S. mansoni miracidia to the snails. Also, it caused a reduction in number of cercariae per snail during the patent period and in the period of cercarial shedding. The results revealed that the glucose concentration in hemolymph and Lactate level in soft tissues of treated snails were increased (P < 0.001) while glycogen, total protein, the lipid content and the pyruvate level in snail’s tissues decreased (P < 0.001). The activities of lactic dehydrogenase (LDH), pyruvatekinase (PK) and cytochrome oxidase (CY) enzymes in homogenate of snail’s tissues were reduced (P < 0.001) in response to treatment with the two tested plants while protease (PR) activity increased (P < 0.001). It is concluded that the application of LC₂₅ of ethanol extract of S. siniacum and A. judaica L. may be helpful in snail control as it interferes with the snails’ biology and physiology.     

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.     

Binary combination of carbohydrates and amino acids as snail attractant in pellets containing molluscicides against the snail Lymnaea acuminata

Snail control is one of the important methods in the campaign to reduce the incidence of fascioliasis and schistosomiasis. In order to achieve this objective, the method of bait formulation containing an attractant and a molluscicide is an appropriate approach to lure the target snail population to the molluscicide. In the present study snail attractant pellets (SAP) were prepared from binary combination of carbohydrates (10 mM) and amino acids (20 mM) in 2% agar solution. These were tested on Lymnaea acuminata, an intermediate host of the digenean trematodes Fasciola hepatica and Fasciola gigantica. The behavioral responses of snails to these binary combination were examined. The fraction of snails that was in contact with the SAP at different times was used as a measure of attraction. Among all the binary combination of carbohydrates; (sucrose + starch)—72.9%, binary combination of amino acids; (proline + serine)—48.0% and binary combination of carbohydrates and amino acids; (sucrose + serine)—69.5%, emerged as the strongest attractant pellets. Toxicity of these SAP containing different concentrations of molluscicides were used as bait against the snail, L. acuminata. Thymol containing SAP emerged as the strongest bait formulation (96 h LC₅₀ 0.540%, 0.318% and 0.305%) against L. acuminata.     

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.