Effect of short term exposure of fenvalerate on biochemical and haematological responses in Labeo rohita (Hamilton) fingerlings

Experiment was carried out to determine the median lethal concentration (LC₅₀) of fenvalerate to Labeo rohita fingerlings. After determining the LC₅₀ value of fenvalerate, a sub-lethal concentration (1/3rd of LC₅₀) of fenvalerate was exposed for 15 days. Significant alterations in SOD (P < 0.05) activity of liver and gill was observed due to fenvalerate. Catalase activity in gills of fishes was also affected significantly (P < 0.05). WBC, NBT and Hct values were reduced significantly in fenvalerate exposed fishes as compared to control group, whereas blood glucose level showed higher values in fenvalerate exposed group. Serum total protein and albumin were also reduced significantly as a result of fenvalerate exposure. Significant increase in the serum GOT, serum GPT, creatinine, triglyceride and serum ACP was noticed after 15 days of fenvalerate exposure. Results indicated that short term exposure of fenvalerate can induce biochemical and haematological alterations causing stress to L. rohita fingerlings.     

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.     

A juvenile hormone analog, pyriproxifen, affects some biochemical components in the hemolymph and fat bodies of Eurygaster integriceps Puton (Hemiptera: Scutelleridae)

Endocrine system has a critical role during the developmental stages of insects by synthesis of several regulatory hormones. One of these hormones is juvenile hormone that several insecticides have been driven based on its biochemical structure e.g. pyriproxifen. Due to various disadvantages of fenitrothione spraying, this study was carried out finding the possible usage of pyriproxifen to control the destructive population of Eurygaster integriceps. After bioassay treatments to acquire the appropriate concentrations, the treatment repeated to find possible changes in the biochemical compounds of hemolymph and reserved macromolecules in fat bodies. Results showed significant discrepancies in amount of biochemical components of the hemolymph and the reserved macromolecules in E. integriceps after pyriproxifen treatment. In hemolymph, activity level of alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase as enzymatic components and uric acid as non-enzymatic one increased but acid phosphatase, lactate dehydrogenase, protein, trehalose and lipid showed adverse results. In fat bodies, the amount of all measured reserves including glycogen, lipid and protein decreased and showed significant differences. These kinds of changes have been supported by several studies due to using insecticides. These negative effects on overall physiology of the Sunn pest by depleting the essential compounds cause sensitivity to fungal infections and several shortages for normal development and reproduction of insect. Also, the adaptability of pyriproxifen to increase the effect of entomopathogenic fungus, Beauveria bassiana, should be considered to initiate a new pest management program to decrease the production loss made by E. integriceps in wheat fields.     

Resistance selection and biochemical characterization of spinosad resistance in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

A Helicoverpa armigera population was collected from Shandong province, China. After 15 generations of selection in the laboratory, the H. armigera strain developed more than 20-fold resistance to spinosad. At LD₅₀ level, no significant cross-resistance was found between spinosad and chlorpyrifos, methomyl, avermectin and chlorfenapyr except for fenvalerate with a low cross-resistance of 2.4-fold. However, LD₉₉ values of fenvalerate against the parental and resistant strains were not different significantly. After inhibitors were used, spinosad resistance could be partially suppressed by piperonylbutoxide (PBO) and triphenylphosphate (TPP), but not by diethylmaleate (DEM). Activities of p-nitroanisole O-demethylase (ODM) developed to 8.26-fold compared with the parental strain, but no obvious changes were found in activities of carboxyl esterase (CarE) and glutathione-S-transferase (GST). The results indicated that resistance to spinosad in the cotton bollworm might be associated with an increase in cytochrome P450 monooxygenase.     

Pythium rot of chingensai ( Brassica campestris L. chinensis group) caused by Pythium ultimum var. ultimum and Pythium aphanidermatum

Severe rot was found at the base of leaves and stems of chingensai (Brassica campestris L. chinensis group) in Okayama Prefecture in 2000. The causal fungi were morphologically identified as Pythium ultimum Trow var. ultimum and P. aphanidermatum (Edson) Fitzpatrick. This is the first report of rot caused by Pythium species on chingensai. We named this disease Pythium rot of chingensai.     

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.     

Growth and physiological responses of freshwater green alga Selenastrum capricornutum to allelochemical ethyl 2-methyl acetoacetate (EMA) under different initial algal densities

Most natural algicides including macrophytic allelochemicals are known to selectively inhibit algal growth. The investigations on the modes of action about the species-specific algicides are little. In this study, the effects of allelochemical ethyl 2-methyl acetoacetate (EMA) identified from reed (Phragmites communis) on the growth, physiological, and biochemical processes of green alga Selenastrum capricornutum were investigated. The results showed that EMA had multiple effects on the growth of S. capricornutum under different initial algal densities (IADs). The algal growth was inhibited by EMA at low IADs, but stimulated at high IADs. Further, the potential modes of action of EMA on S. capricornutum were explored from ultrastructure, metabolic activity, reactive oxygen species level, and lipid peroxidation to trace the microenvironment changes in the algal cells. Damage in cell structure occurred at low IAD, but cells were well developed with increased metabolic activity at high IAD. The reactive oxygen species (ROS) levels were increased under both conditions. The increase of ROS level was acute at low IAD but slow at high IAD. EMA caused significant lipid peroxidation, i.e. oxidative damage on membrane lipids at low IAD but not at high IAD. Based on these results, the initial algal density is considered an important factor to influence algal growth and physiological and biochemical responses to EMA, the effects of EMA on S. capricornutum may be “hormesis-like”, and different ROS increase ratio may be directly related with different responses of S. capricornutum to EMA.     

Induction of an antioxidant enzyme system and other oxidative stress markers associated with compatible and incompatible interactions between chickpea (Cicer arietinum L.) and Fusarium oxysporum f. sp.ciceris

To ascertain if active oxygen species play a role in fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceris, the degree of lipid peroxidation (malondialdehyde formation) and the activity levels of diamine oxidase (DAO), an apoplastic H₂O₂-forming oxidase, and several antioxidant enzymes, namely ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR), guaiacol-dependent peroxidase (GPX) and superoxide dismutase (SOD), were determined spectrophotometrically in roots and stems of ‘WR315’ (resistant) and ‘JG62’ (susceptible) chickpea cultivars inoculated with the highly virulent race 5 of the pathogen. Moreover, APX, CAT, GPX and SOD were also analysed in roots and stems by gel electrophoresis and activity staining; and the protein levels of APX and SOD in roots were determined by Western blotting. In roots, infection by the pathogen increased lipid peroxidation and CAT and SOD activities, although such responses occurred earlier in the incompatible compared with the compatible interactions. APX, GPX and GR activities were also increased in infected roots, but only in the compatible interaction. In stems, infection by the pathogen increased lipid peroxidation and APX, CAT, SOD and GPX activities only in the compatible interaction, and DAO activity only in the incompatible one. In general, electrophoregrams agreed with the activity levels determined spectrophotometrically and did not reveal any differences in isoenzyme patterns between cultivars or between infected and non-infected plants. Further, Western blots revealed an increase in the root protein levels of APX in the compatible interaction and in those of SOD in both compatible and incompatible interactions. In conclusion, whereas enhanced DAO activity in stems, and earlier increases in lipid peroxidation and CAT and SOD activities in roots, can be associated with resistance to fusarium wilt in chickpea, the induction of the latter three parameters in roots and stems along with that of APX, GR (only in roots) and GPX (only in stems) activities are rather more associated with the establishment of the compatible interaction.     

Effects of three esca-associated fungi on Vitis vinifera L.: I. Characterization of secondary metabolites in culture media and host responses to the pathogens in calli

Phaeomoniella chlamydospora (Pch) and Togninia minima (Tmi) produced scytalone, isosclerone and pullulans in liquid cultures, as well as in calli. Secondary metabolites and host defense compounds were shown to occur in calli of Vitis vinifera cv. Italia and cv. Matilde infected by the esca-associated fungi Pch, Tmi and Fomitiporia mediterranea (Fme). Calli of both cvs. were grown as dual cultures with Pch, Tmi and Fme. The fungi grew well in the presence of calli of both cvs., but callus growth was reduced. Accumulation and changes of total phenolics and recurring phenolics, and of two phytotoxic pentaketides and pullulans were analyzed by HPLC. On comparing results for cv. Italia and cv. Matilde, it can be seen that concentrations of phenolics are strongly related to the cv. The paper discusses the possible relationship between melanin biosynthesis in Pch and Tmi, which utilize pentaketide metabolites as intermediates and their pathogenicity related to phytotoxity of scytalone and isosclerone.     

Toxicological and metabolic parameters of the teleost fish (Leporinus obtusidens) in response to commercial herbicides containing clomazone and propanil

Pesticides, such as herbicides can affect the metabolic and toxicological parameters on fish. For this reason, an experiment was carried out with the objective of to evaluate the effects of commercial formulations of clomazone and propanil herbicides on acetylcholinesterase (AChE), thiobarbituric acid-reactive substances (TBARS), catalase (CAT) and metabolic parameters in teleost fish (Leporinus obtusidens). Fish were exposed during 90 days to field measured concentration of the herbicides clomazone and propanil (376 and 1644 μg/L, respectively) on rice paddy water. Specific AChE activity in the brain and muscle decreased and TBARS levels decreased in brain, muscle and liver tissues. Liver catalase decreased after exposure to both herbicides. Metabolic parameters in the liver and white muscle showed different changes after exposure to both herbicides. In summary, the results showed that clomazone and propanil affects toxicological and metabolic parameters of piavas. These results suggest that environmentally relevant herbicides concentrations are toxic to Leporinus obtusidens.     

Growth, photosynthetic pigments and photosynthetic activity during seedling stage of cowpea (Vigna unguiculata) in response to UV-B and dimethoate

UV-B (0.4 W m⁻²) irradiation and dimethoate (100 and 200 ppm) treatments, singly and in combination, declined the growth, photosynthetic pigment contents and photosynthesis (O₂ evolution and CO₂-fixation) of cowpea (Vigna unguiculata). Contrary to this, low concentration of dimethoate (50 ppm) caused stimulation on these parameters, while together with UV-B it showed inhibitory effects. Carotenoids (Car) showed varied responses. It was found that carbon-fixation (¹⁴CO₂) was more sensitive to both the stresses than photosynthetic oxygen evolution. Photosynthetic electron transport activity was reduced by both the stresses, however, 50 ppm dimethoate besides inhibiting photosystem II (PSII) and whole chain activity, showed slight stimulation in photosystem I (PSI) activity. The individual effect of two stresses on PSII activity was probably due to interruption of electron flow at oxidation side of PSII which extended to its reaction center following simultaneous exposure. A similar trend was also noticed in case of CO₂ liberation (measured as ¹⁴CO₂ release) in light and dark. Results suggest that dimethoate (100 and 200 ppm) and UV-B alone caused heavy damage on pigments and photosynthetic activity of cowpea, leading to the significant inhibition in growth. Further, the interactive effects of both the stresses got intensified. However, low concentration (50 ppm) of dimethoate showed stimulation, but in combination, it slightly recovered from the damaging effect, caused by UV-B.     

Insecticide resistance status of the codling moth Cydia pomonella (Lepidoptera: Tortricidae) from Greece

The codling moth Cydia pomonella L. is controlled mostly with chemical insecticides in Greece and control failures have been reported. However, there are no insecticide resistance studies in the country as yet. We examined the insecticide resistance status of 33 and 38 populations of fifth-instar non-diapausing and diapausing larvae, respectively by applying bioassays, biochemical and DNA diagnostics. Diagnostic concentrations of azinphos-methyl, phosalone, deltamethrin, thiacloprid, fenoxycarb, tebufenozide, methoxyfenozide and diflubenzuron were used in bioassays. Almost all populations showed reduced susceptibility to at least one insecticide and approximately half of them to all insecticides examined compared to a laboratory susceptible strain used as reference. However, only one out of six populations tested showed reduced susceptibility in ovicidal tests with fenoxycarb. Cross-resistances were observed among most insecticides, except from the pairs fenoxycarb–phosalone and thiacloprid–phosalone, in non-diapausing larvae. The more obvious biochemical marker associated with the reduced susceptibility observed in both larval instars was elevated cytochrome P₄₅₀ polysubstrate monooxygenases activity, followed by elevated glutathione-S-transferase activity and reduced carboxylesterases activity. Neither sodium channel nor AChE known resistance mutations were found in any of the approximately 1000 individuals of each larval instar screened with diagnostic PCR. Actions for Integrated Resistance Management and application of alternative control methods are discussed.     

Biological, biochemical, and molecular parameters of Helisoma duryi snails exposed to the pesticides Malathion and Deltamethrin

Aquatic environments get contaminated with pesticides residues that result from the application of pesticides in agricultural practices. The present study aimed to evaluate the pesticides Malathion and Deltamethrin on biological and biochemical parameters of Helisoma duryi snails. The results showed that LC₁₀ of the two pesticides caused considerable reduction in survival rates and egg production of treated snails. Glucose concentration in hemolymph of the exposed snails showed a significant increase. On the other hand, albumin in hemolymph and glycogen contents as well as the activities of enzymes in tissues of snails including lactate dehydrogenase (LDH), succinate dehydrogenase (SDH), arginase, and ornithine aminotransferase (OAT) were significantly inhibited. Meanwhile, the activity of AMP deaminase of the exposed snails was significantly increased in the hemolymph in response to treatment. The electrophoretic pattern of total protein showed differences in number and molecular weights of protein bands. DNA concentration was investigated by measuring the intensity of the genomic bands and its showed its increase in the treated snails. It was concluded that the residues of Malathion and Deltamethrin pesticides in aquatic environments have toxic effects on non-target organism, e.g. H. duryi snails.     

Induction of hypericins and hyperforin in Hypericum perforatum L. in response to biotic and chemical elicitors

Hypericum perforatum L. produces hyperforins, a family of antimicrobial acylphloroglucinols; and hypericins, a family of phototoxic anthraquinones exhibiting anti-microbial, anti-viral, and anti-herbivore properties in vitro. To determine whether these secondary metabolites are part of the specific plant defense systems that are mediated by methyl jasmonate or salicylic acid, we used meristem cultures to assess the effects of exposure to exogenous application of these chemical elicitors. Levels of hypericins in plant tissue increased in response to both elicitor treatments; total hypericin levels increased as much as 3.3 times control levels when treated with 200 μ methyl jasmonate for 14 days. Increased hyperforin concentrations were detected when plantlets were treated with 1 m salicylic acid or 50 μ methyl jasmonate. For assessing responses to a biotic elicitor, greenhouse-grown plant materials were inoculated with the plant pathogen, Colletotrichum gloeosporioides. Levels of hypericins increased twice as much as the control when inoculated with 1 × 10⁴ spores per ml; higher doses of spores overwhelmed the plant defenses. The elevation of hypericins and hyperforin in response to chemical and biotic elicitors suggests that these secondary metabolites are components in the inducible plant defense responses of H. perforatum.     

In vitro and in vivo effects of some pesticides on glucose-6-phosphate dehydrogenase enzyme activity from rainbow trout (Oncorhynchus mykiss) erythrocytes

We investigated the in vitro and in vivo effects of some pesticides on rainbow trout erythrocyte glucose-6-phosphate dehydrogenase enzyme. The enzyme was purified 1691-fold with a specific activity of 16.235 U/mg protein and a yield of 63%. Cypermethrin, and propoxur inhibited glucose-6-phosphate dehydrogenase enzyme in vitro and deltamethrin inhibited both in vivo and in vitro. The obtained IC₅₀ values for deltamethrin, cypermethrin, and propoxur were 0.63, 1.02, and 12 mM, respectively. The activity of the control was determined as 5.17 ± 0.62 U/g Hb in in vivo studies. The enzyme activities of the groups treated with 0.25 g/L deltamethrin were measured at 6, 12, 24, 48, and 72 h, and found to be 4.32 ± 0.47, 3.57 ± 0.39, 3.47 ± 0.45, 2.86 ± 0.37, and 2.31 ± 0.32 U/g Hb. In vivo experiments showed that deltamethrin significantly inhibited the G6PD enzyme activity after the 48th h (p < 0.05).     

Metabolic resistance mechanisms to phosalone in the common pistachio psyllid, Agonoscena pistaciae (Hem.: Psyllidae)

The common pistachio psyllid, Agonoscena pistaciae, is the most damaging pest of pistachio in Iran, and is generally controlled by insecticides belonging to various classes especially, phosalone. The toxicity of phosalone in nine populations of the pest was assayed using the residual contact vial and insect-dip methods. The bioassay results showed significant discrepancy in susceptibility to phosalone among the populations. Resistance ratio of the populations to the susceptible population ranged from 3.3 to 11.3. The synergistic effects of TPP, PBO and DEM were evaluated on the susceptible and the most resistant population to determine the involvement of esterases, mixed function oxidases and glutathione S-transferases in resistance mechanisms, respectively. The level of resistance to phosalone in the resistant population was suppressed by TPP, PBO and DEM, suggesting that the resistance to phosalone is mainly caused by esterase detoxification. Biochemical enzyme assays revealed that esterase, glutathione S-transferase and cytochrome P450 monooxygenase activities in the resistant population was higher than that in the susceptible. Glutathione-S-transferases play a minor role in the resistance of the pest to phosalone.     

Impact of sugarcane yellow leaf virus (ScYLV) on the carbohydrate status of sugarcane: Comparison of virus-free plants with symptomatic and asymptomatic virus-infected plants

The carbohydrate concentration of old and young sugarcane plants, either virus-free or infected by sugarcane yellow leaf virus, was compared. Internode lengths and fresh weights of symptomatic plants were reduced, but not the sucrose concentrations. Starch and sucrose concentration in green leaf tops of mature (18 months old) was the highest in asymptomatic plants followed by symptomatic, and then by virus-free plants. Carbohydrates increased during daytime by 5–20%. The carbohydrate concentration in leaves of young, 6 months old plants was much lower than in mature plants and it increased to 500% during daytime. Asymptomatic leaves had a higher level of carbohydrates, especially starch, from late afternoon until the end of the night, suggesting a reduction of assimilate export.     

Histochemical studies on the accumulation of reactive oxygen species (O2 and H2O2) in the incompatible and compatible interaction of wheat—Puccinia striiformis f. sp. tritici

The generation and accumulation of reactive oxygen species (ROS), superoxide anion (O₂⁻) and hydrogen peroxide (H₂O₂), were studied in the interaction between wheat cv. ‘Suwon 11’ and two races of Puccinia striiformis f. sp. tritici (avirulent and virulent). Generation of O₂⁻ and H₂O₂ was analyzed histochemically using nitroblue tetrazolium (NBT) and 3,3-diamino-benzidine (DAB), respectively. At the pre-penetration stage during appressorium formation both stripe rust races induced H₂O₂ accumulation in guard cells. In the incompatible interaction, a rapid increase of O₂⁻ and H₂O₂ generation at infection sites was detected. The percentage of infection sites showing NBT and DAB staining was 36.1% and 40.0%, respectively, 12 h after inoculation (hai). At extended incubation time until 24 hai, percentage of infection sites showing H₂O₂ accumulation further increased, whereas those exhibiting O₂⁻ accumulation declined. The early infection stage from 12 to 24 hai coincided with primary haustoria formation in mesophyll cells. In contrast, in the compatible interaction, O₂⁻ and H₂O₂ generation could not be detected in most of the infection sites. In the incompatible interaction, intensive DAB staining was also determined in mesophyll cells, especially in cell walls, surrounding the infected cells 16–24 hai; thereafter, these cells contained fluorescing compounds and underwent hypersensitive response (HR). The number of necrotic host cells surrounding the infection sites increased continuously from 20 to 96 hai. It might be concluded that H₂O₂ accumulation during the early infection stage is associated with the occurrence of hypersensitive cell death and that resistance response is leading to arrest the avirulent race of the obligate stripe rust pathogen. In the compatible interaction at 96 hai, H₂O₂ accumulation was observed in mesophyll cells surrounding the rust lesion.