Green nano-emulsion intervention for water-soluble glyphosate isopropylamine (IPA) formulations in controlling Eleusine indica (E. indica)

This article describes the development of environmentally friendly nano-emulsion system for water-soluble herbicide application. Pseudoternary phase diagrams were established in the emulsion system of fatty acid methyl esters (FAMEs)/alkylpolyglucosides (APG) and/or 3-(3-hydroxypropyl)-heptamethyltrisiloxane (organosilicone)/water encompassed with 41% (w/w) glyphosate isopropylamine (IPA) as herbicide active. Pre-formulations were selected from isotropic (L) region in the phase diagrams and their emulsion system characteristics were determined. The microemulsion systems were chosen and then dispersed into water using low-energy stirring method (200 rpm for 5 min). Oil-in-water (O/W) nano-emulsions were formed with particle sizes of diameter less than 200 nm. The nano-emulsion systems showed significantly lower surface tension than a commercial formulation (Roundup®). In the biological application study, treatments of nano-emulsion formulations and Roundup® were applied on narrow-leaved weed Eleusine indica. Multiple doses of glyphosate IPA of the treatments were applied for the construction of dose-response curves for determination of effective dose (ED₅₀). The nano-emulsion formulation showed lower ED₅₀ was 0.40 kg a.e./ha in controlling the weed than Roundup® was 0.48 kg a.e./ha. This finding suggested that the possibility of using nano-emulsion system to increase penetration and uptake of glyphosate IPA.     

Multiple effects of a naturally occurring proline to threonine substitution within acetolactate synthase in two herbicide-resistant populations of Lactuca serriola

Two populations of Lactuca serriola L. with resistance to acetolactate synthase (ALS)-inhibiting herbicides were discovered in wheat fields at two locations more than 25 km apart in South Australia. Both resistant populations carried a single base change within a highly conserved coding region of the ALS gene that coded for a single amino acid modification within ALS. The modification of proline 197 to threonine resulted in an enzyme that was highly resistant (>200-fold) to inhibition by sulfonylurea herbicides and moderately resistant to triazolopyrimidine and imidazolinone herbicides. The herbicide-resistant ALS was also less sensitive to inhibition by the branched-chain amino acids valine and leucine. In addition, the resistant enzyme had a lower K_m for pyruvate. However, extractable ALS activity was similar between resistant and susceptible plants. The substitution of threonine for proline 197 within ALS has multiple impacts on ALS enzyme activity in L. serriola that may influence the frequency of this resistant allele in the environment.     

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.     

Bio-potency of serine proteinase inhibitors from Acacia senegal seeds on digestive proteinases, larval growth and development of Helicoverpa armigera (Hübner)

Proteinase inhibitors (AsPIs) with high activity against serine proteinases were purified from seeds of the tree legume, Acacia senegal by ammonium sulfate precipitation followed by DEAE-Sephadex A-25 column and evaluated against Helicoverpa armigera larvae by in vitro and in vivo methods. The molecular weight of AsPIs was found to be approximately 19.58 ± 1.00 and 21.23 ± 1.00 kDa for PI and 18.16 ± 1.00 kDa for PII on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The AsPIs (5 μg/ml) inhibited approximately 70% of midgut trypsin and 61% of elastase-like chymotrypsin. In vitro studies showed that AsPIs have remarkable inhibitory activity towards total gut proteolytic enzymes followed by trypsin and chymotrypsin. The IC₅₀ of AsPIs for midgut trypsin was 0.1 μg/ml and for chymotrypsin was 2.0 μg/ml. The inhibition of gut proteinase enzymes was of the non-competitive type. In larval feeding studies, AsPIs were found to retard growth and development of H. armigera and also affects the fecundity of the pest. The results advocate the use of AsPIs in transgenic technology to develop plant resistance to H. armigera.     

Inhibitory effects of Schiff analogs of salicylidene aniline on phenoloxidase from Pieris rapae L. (Lepidoptera: Pieridae)

In order to gain insight into the development of insecticides with novel modes of action, the effects of salicylidene aniline (a), salicylidene-4-chloroaniline (b), salicylidene-4-bromoaniline (c), and salicylidene-4-nitroaniline (d) on partially purified phenoloxidase (PO) from Pieris rapae L. were investigated. The results showed that the 4 compounds could inhibit PO activity, and the inhibitor concentrations leading to a loss of 50% activity (IC₅₀) were estimated to be 0.025 mmol L⁻¹, 0.732 mmol L⁻¹, 0.471 mmol L⁻¹, and 0.675 mmol L⁻¹, respectively. Meanwhile, all the inhibitors showed reversible competitive inhibition, except (d), which showed reversible mixed inhibition. The K_I values were determined as 0.106 mmol L⁻¹, 10.059 mmol L⁻¹, 8.390 mmol L⁻¹, and 20.198 mmol L⁻¹ for the four compounds, respectively. The UV-vis spectra of (a) and (d) in the presence of copper ions and the enzyme showed that (a) could directly chelate the copper ions of PO; however, (d) could neither chelate the additional copper ions nor the copper ions of PO.     

Cloning of the acetylcholinesterase 1 gene and identification of point mutations putatively associated with carbofuran resistance in Nilaparvata lugens

Molecular mechanisms of carbofuran resistance in the brown planthopper, Nilaparvata lugens Stål, were investigated. A carbofuran-resistant strain (CAS) showed approximately 45.5- and 15.1-fold resistance compared with a susceptible strain (SUS) and a non-selected field strain (FM), respectively. Activities of the esterase and mixed-function oxidase were approximately 2.8- and 1.6-fold higher, respectively, in the CAS strain than in the SUS strain, suggesting that these enzymes play a minor role in carbofuran resistance. Interestingly, the insensitivity of acetylcholinesterase (AChE) to carbofuran was approximately 5.5- and 3.7-fold higher in the CAS strain compared to the SUS and FM strains, respectively, indicating that AChE insensitivity is associated with carbofuran resistance. Western blot analysis identified two kinds of AChEs, of which the type-1 AChE (encoded from Nlace1, which is paralogous to the Drosophila AChE gene) was determined to be the major catalytic AChE in N. lugens. The open reading frame of Nlace1 is composed of 1989 bp (approximately 74 kD) and revealed 52.5% and 24.3% amino acid sequence identities to those of Nephotettix cincticeps and Drosophila melanogaster, respectively. Screening of point mutations identified four amino acid substitutions (G119A, F/Y330S, F331H and H332L) in the CAS strain that likely contribute to AChE insensitivity. The frequencies of these mutations were well correlated with resistance levels, confirming that they are associated with reduced sensitivity to carbofuran in N. lugens. These point mutations can be useful as genetic markers for monitoring resistance levels in field populations of N. lugens.     

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.     

In vitro inhibition of Sclerotinia sclerotiorum by mixtures of azoxystrobin, SHAM, and thiram

The necrotrophic fungal phytopathogen Sclerotinia sclerotiorum (Lib.) de Bary has a broad host range and frequently causes destructive diseases. The extensive use of common fungicides to control these diseases has selected for resistance in populations of S. sclerotiorum. In this study, 105 isolates of S. sclerotiorum from different geographical regions in Jiangsu Province of China were characterized for baseline sensitivity to azoxystrobin, and the average EC₅₀ value was 0.2932 μg/mL for mycelial growth. Of the mixtures of the fungicides thiram and azoxystrobin that were tested using an in vitro mycelial growth assay, the 1:4 ratio provided the greatest inhibition of S. sclerotiorum. When tested against nine isolates, the 1:4 mixture resulted in a mean synergy ratio of 2.31, indicating synergistic inhibition. Mycelial respiration was inhibited for about 2 h by azoxystrobin alone but for 48 h by the mixture of thiram and azoxystrobin. Salicylhydroxamic acid (SHAM, a known inhibitor of alternative respiration) also increased the inhibition of mycelial growth and respiration caused by azoxystrobin. These results suggest the need for further study of effects of combinations of azoxystrobin with thiram or SHAM in planta to evaluate their potential for management of diseases caused by S. sclerotiorum.     

Purification and partial characterization of a glutathione S-transferase from the two-spotted spider mite, Tetranychus urticae

Glutathione S-transferases (GSTs) are known to catalyze conjugations by facilitating the nucleophilic attack of the sulfhydryl group of endogenous reduced glutathione on electrophilic centers of a vast range of xenobiotic compounds, including insecticides and acaricides. Elevated levels of GSTs in the two-spotted spider mite, Tetranychus urticae Koch, have recently been associated with resistance to acaricides such as abamectin [Pestic. Biochem. Physiol. 72 (2002) 111]. GSTs from acaricide susceptible and resistant strains of T. urticae were purified by glutathione-agarose affinity chromatography and characterized by their Michaelis-Menten kinetics towards artificial substrates, i.e., 1-chloro-2,4-dinitrobenzene and monochlorobimane. The inhibitory potential of azocyclotin, dicumarol, and plumbagin was low (IC₅₀ values > 100 μM), whereas ethacrynic acid was much more effective, exhibiting an IC₅₀ value of 4.5 μM. GST activity is highest in 2-4-day-old female adults and dropped considerably with progressing age. Furthermore, molecular characteristics were determined for the first time of a GST from T. urticae, such as molecular weight (SDS-PAGE) and N-terminal amino acid sequencing (Edman degradation). Glutathione-agarose affinity purified GST from T. urticae strain WI has a molecular weight of 22.1 kDa. N-terminal amino acid sequencing revealed a homogeneity of ≈50% to insect GSTs closely related to insect class I GSTs (similar to mammalian Delta class GSTs).     

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.     

Resistance development in rice blast disease caused by Magnaporthe grisea to tricyclazole

Sensitivity to tricyclazole of 129 single-conidial isolates of rice blast fungus, Magnaporthe grisea, was determined. EC₅₀ values ranged from 0.06 to 1.12 mg/L with an average value of 0.46 ± 0.09 mg/L according to the detached leaf segment tests. No significant difference of sensitivity was observed between isolates from Guangdong and Jiangsu where decreased efficacy was reported, and from two other provinces where tricyclazole provided excellent disease control. In seedling tests, tricyclazole could control the most tolerant isolate GY-6 successfully with a efficacy of 81.5% at the concentration of 40 mg/L. Sensitivities of GY-6 and DY-2, the most sensitive isolate, to tricyclazole were both unstable in sub-cultured single-conidial offspring isolates, with respective mean EC₅₀ values of 5.40 ± 0.97 and 4.50 ± 0.88 mg/L calculated from seedling tests. There was no amino acid difference between them in the coding sequences of 1,3,6,8-tetrahydroxynaphthalene reductase and 1,3,8-trihydroxynaphthalene reductase. These results suggested that the decreased control reported in Guangdong and Jiangsu could not be attributed to the occurrence of resistance. When continuously “inoculated-reisolated-reinoculated” under the selection of tricyclazole in vivo, sensitivity of DY-2 decreased 10-fold after 20 generations, although the sensitivity of GY-6 did not shift significantly.     

In vitro and in vivo effects of some pesticides on carbonic anhydrase enzyme from rainbow trout (Oncorhynchus mykiss) gills

Here we investigated the in vitro and in vivo effects of the pesticides, deltamethrin, diazinon, propoxur and cypermethrin, on the activity of rainbow trout (rt) gill carbonic anhydrase (CA). The enzyme was purified from rainbow trout gills using Sepharose 4B-aniline-sulfanilamide affinity chromatography method. The overall purification was approx. 214-fold. SDS-polyacrylamide gel electrophoresis showed a single band corresponding to a molecular weight of approx. 29 kDa. The four pesticides dose-dependently inhibited in vitro CA activity. IC₅₀ values for deltamethrin, diazinon, propoxur and cypermethrin were 0.137, 0.267, 0.420 and 0.460 μM, respectively. In vitro results showed that pesticides inhibit rtCA activity with rank order of deltamethrin > diazinon > propoxur > cypermethrin. Besides, in vivo studies of deltamethrin were performed on CA activity of rainbow trout gill. rtCA was significantly inhibited at three concentrations (0.25, 1.0 and 2.5 μg/L) at 24 and 48 h.     

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.     

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.     

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.     

山东省部分市县麦田杂草麦家公Lithospermum arvense对苯磺隆的抗药性

采用温室盆栽法和培养皿法测定了山东省部分市县冬小麦田杂草麦家公Lithospermum arvense L.对苯磺隆的抗药性水平,以及其抗药性生物型乙酰乳酸合成酶(ALS)对苯磺隆的敏感性。温室盆栽结果显示,供试杂草对苯磺隆产生了不同程度的抗药性,其中胶州麦家公生物型抗性水平最高,抗性倍数为12.8倍;培养皿法测定结果也显示胶州麦家公生物型抗性水平最高,但抗性倍数为3.89倍。交互抗性测定结果表明,胶州抗性麦家公生物型对其他ALS抑制剂噻吩磺隆和苄嘧磺隆已产生不同程度的交互抗性,其中对噻吩磺隆的抗性倍数达到3.11倍。离体条件下,与敏感生物型ALS活力的抑制中浓度(IC50)相比较,胶州抗性麦家公生物型的IC50值是敏感麦家公的 2.65倍。表明ALS敏感性降低可能是山东部分市县麦家公对苯磺隆产生抗药性的重要原因之一。     

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.     

Brusatol isolated from Brucea javanica (L.) Merr. induces apoptotic death of insect cell lines

Brucea javanica (L.) Merr. is a medicine plant distributed widely throughout Asia where its bitter fruits have been used traditionally in medicine for treating various ailments and controlling some pests. In recent years, concerns over the potential impact of synthetic pesticides on human health and environment have now become more pressing to develop environmentally friendly pesticides. In this paper, brusatol, a quassinoid, was isolated from the fruit of B. javanica, and identified using X-ray crystallographic analysis. Results showed that brusatol has potent contact toxicity (LD₅₀, 2.91 μg/larva, 72 h) and anfieedant activity (AFC₅₀, 17.4 mg/L, 48 h) against the third-instar larvae of Spodoptera exigua. Brusatol demonstrated cytotoxic effects to the tested insect cell lines, IOZCAS-Spex-II and Sf21, in a time- and dose-dependent manner. After brusatol treatment, apoptotic cell death with the DNA fragmentation, activation of caspase-3 and release of cytochrome c was preliminarily observed in both IOZCAS-Spex-II and Sf21. These results indicated the existence of apoptotic death with the mitochondrial-dependent pathway induced by brusatol in Sf21 and IOZCAS-Spex-II cell lines. Our studies will provide important knowledge to understand mechanisms of action of brusatol and to develop brusatol and its derivatives as insecticides.     

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.     

Insecticide resistance in the tropical bedbug Cimex hemipterus

Insecticide resistance in the bedbug Cimex hemipterus was investigated using 4211 bedbugs collected from three districts of Sri Lanka. Insecticide bioassays were carried out with discriminating dosages of deltamethrin, permethrin, DDT, malathion, and propoxur. Activity levels of insecticide metabolizing enzymes and the insecticide target site acetylcholinesterase were monitored using biochemical assays. Percentage survivals after DDT, malathion, and propoxur exposure were 41-88%, 18-64%, and 11-41%, respectively. For deltamethrin and permethrin, KT₅₀/KT₉₀ (time to knock-down 50%/90% of the population) values were 0.5-24/1.0-58 and 1.3-10/2.5-47 h, respectively. Both elevated esterase and malathion carboxylesterase mechanisms were present in bedbug populations. Monooxygenase levels were heterogeneous. Organophosphate and carbamate target site acetylcholinesterase, was insensitive in 29-44% of the populations. High DDT resistance was probably due to glutathione S-transferases. Malathion carboxylesterases are mainly responsible for high malathion resistance. High tolerance to both DDT and pyrethroids suggests the presence of ‘kdr’ type resistance mechanism in one population.