Elevated Esterase Activity Research Articles

Insecticide Susceptibilities and Enzyme Activities of Four Stink Bug Populations in Mississippi, USA.

In Mississippi, the Pentatomidae complex infesting soybean is primarily composed of Euschistus servus, Nezara viridula, Chinavia hilaris, and Piezodorus guildinii. This study employed spray bioassays to evaluate the susceptibilities of these stink bugs to seven commonly used formulated insecticides: oxamyl, acephate, bifenthrin, λ-cyhalothrin, imidacloprid, thiamethoxam, and sulfoxaflor. Stinks bugs were collected from soybeans in Leland, MS, USA during 2022 and 2023, as well as from wild host plants in Clarksdale, MS. There was no significant difference in the susceptibility of C. hilaris to seven insecticides between two years, whereas P. guildinii showed slightly increased susceptibility to neonicotinoids in 2023. Among all four stink bug species, susceptibility in 2022 was ranked as P. guildinii ≤ C. hilaris ≈ N. viridula, while in 2023, it was ranked as P. guildinii ≤ C. hilaris ≤ E. Servus. Additionally, populations of E. servus and P. guildinii collected from Clarksdale exhibited high tolerance to pyrethroids and neonicotinoids. Moreover, populations of E. servus and P. guildinii from SIMRU-2022 and Clarksdale-2023 showed elevated esterase and cytochrome P450 activity, respectively. These findings from spray bioassays and enzyme activity analyses provide a baseline for monitoring insecticide resistance in Pentatomidae and can guide insecticide resistance management strategies for Mississippi soybean.

Biochemical and molecular characterization of neonicotinoids resistance in the tarnished plant bug, Lygus lineolaris

In the southern United States, neonicotinoids are commonly applied as foliar insecticides to control sucking insect pests, such as the tarnished plant bug (TPB, Lygus lineolaris). In this study, spraying bioassays were conducted to determine the toxicity of five neonicotinoids and sulfoxaflor to susceptible and late fall field-collected TPB adults from Mississippi Delta region. Compared to a susceptible population, the field-collected TPBs exhibited the highest resistance to imidacloprid (up to 19.5-fold), a moderate resistance to acetamiprid (9.43-fold), clothianidin (13.68-fold), thiamethoxam (7.88-fold) and the least resistance to thiacloprid (4.61-fold) and sulfoxaflor (1.82-fold), respectively. A synergist study demonstrated that piperonyl butoxide (PBO) significantly increased the toxicity of imidacloprid and thiamethoxam by 22.2- and 15.3-fold, respectively, while triphenyl phosphate (TPP) and diethyl maleate (DEM) only showed 2–3-fold synergism to both neonicotinoids. In the field-collected TPBs, activities of the three detoxification enzymes esterase, glutathione S-transferase (GST) and CYP450 monooxygenase (P450) were significantly increased by 3.43-, 1.48- and 2.70-fold, respectively, when compared to the susceptible population. Additionally, after 48 h exposure to imidacloprid or thiamethoxam, resistant TPB adults exhibited elevated esterase activities, decreased GST activities, and no significant changes in P450 activities. Further examinations revealed that the expression of certain esterase and P450 detoxification genes were significantly elevated in resistant TPBs. Overall, these results suggest that elevated esterase and P450s expression and enzyme activity are key mechanisms for metabolic resistance in TPBs to neonicotinoids. Our findings also provide valuable information for selection and adoption of neonicotinoid insecticides for resistance management of TPBs and minimizing toxic risk to foraging bees.

Duloxetine ameliorates valproic acid-induced hyperactivity, anxiety-like behavior, and social interaction deficits in zebrafish.

Syndromic autism spectrum disorders (ASDs) are characterized by impaired social communication and repetitive/stereotyped behaviors. Currently available therapeutic agents against ASD have limited efficacy. Thus, searching for novel and effective drugs ameliorating core symptoms, in particular social deficits, is of utmost importance. Duloxetine (DLX), an antidepressant that has been identified as an agonist mimetic for the cell adhesion molecule L1, exhibits beneficial functions in vitro and in vivo. Therefore, in this study, we focused on the rapid and persistent neuroprotective function of DLX following valproic acid (VPA)-triggered hyperactivity, anxiety-like behavior and social deficits in zebrafish. Embryonic exposure to VPA reduced survival in a dose- and time-dependent manner, delayed hatching, and also resulted in a significant number of malformed larvae. After initial dose-response experiments in zebrafish larvae, 10μM VPA exposure between 0.33 and 4.5 days post fertilization (dpf) was identified as an effective concentration that led to an early and persistent ASD-like phenotype in zebrafish. ASD-like elevated acetylcholine esterase (AChE) activity and reduced Akt-mTOR signaling was observed in zebrafish whole brain. Acute administration of DLX (4.5-6 dpf) reduced the VPA-induced ASD-like phenotype in zebrafish larvae. Additionally, such early-life acute DLX treatment had long-term effects in ameliorating social impairments, hyperactivity, and anxiety-like behaviors through adulthood. This was accompanied by reduced AChE activity and by normalized Akt-mTOR signaling. Overall, DLX treatment showed a long-term therapeutic effect on autistic-like behaviors, and alteration of AChE activity and Akt-mTOR signaling were identified as crucial in the VPA-induced ASD zebrafish model.

A non-contact device for fast screening of wound infections.

Screening for wound infection relies on the expertise of the provider. Clinical diagnosis of infections based on wound swab/biopsy results often takes a few days and may not assess the full wound. There is a need for a non-invasive tool that can quickly and accurately diagnose wound infection. Leukocyte esterase strips are used to identify various infectious diseases. However, it is not clear whether infected wounds also have elevated leukocyte esterase activities as compared with non-infected wounds. To achieve the objective, a device was developed to detect elevated leukocyte esterase activities in wounds by measuring wound exudates adsorbed onto wound dressings in 3minutes. The efficacy of the device in assessing leukocyte esterase activities across various chronic wounds was tested. Such measurements were unaffected by the type of underlying wound dressing. By correlating the device outputs with clinical adjudication of infection, we found that this device had high positive predictive values for diagnosing wound infection in a wide variety of chronic wounds. In addition, a positive device output increases the probability of detecting infected wounds, while the negative device output reduces the probability of detecting infected wounds. This rapid non-contact and disposable diagnostic tool may serve as a rapid and accurate indication of infection in the chronic wound.

Uji mikroplat aktivitas enzim esterase untuk mendeteksi resistensi Anopheles aconitus terhadap insektisida organofosfat

Non specific esterase are known to be important detoxification enzyme contributing to development of insecticides resistance in mosquitoes. Elevated esterase activity is one of the mechanisms of resistance to organophosphate, carbamate and pyrethroid insecticides in the mosquitoes. Such metabolic resistance can be detected by a microplate assay method. Mosquitoes vector resistance can occurs as a result of continuous exposure to the insecticide. Organophosphate insecticide has been used in the vector control program on Anopheles. aconitus, the malaria vector in Jepara Regency since 1983. The use of organophosphate and carbamate insecticides for five years for controlling Anopheles nigerrimus in Srilanka contribute to the selective resistance. This could happen to the population of An. aconitus in Jepara Regency. The objectives of this studies was to determine the potency of An. aconitus from Jepara Regency to be resistant to organophosphate insecticide related to esterase activity mechanism. The study methods used was biochemical assays (microplate assays )for elevated esterase. The level of esterases in larvae was determined using a and b naphthyl acetate, as the substrate andFast Blue B as the coupling reagent. The esterase activity was measured at 450 nm with Dytech ELISA plate reader. Microplate assay (Biochemical assays) on individual An. aconitus from Mlonggo II and Bangsri III subdistrics revealed that 12,9% and 28,6% population was resistant to organophosphate insecticide. The result showed that there was significant difference of elevated of esterase activity with both a and b naphthylacetate substrate hydrolysis, which appeared to be the major resistance mechanism in this multiple organophosphate resistant strain. Therefore the use of another insecticide group for vector control (An. aconitus) was suggested.

Feeding and digestion of the marine isopod Idotea emarginata challenged by poor food quality and microplastics

Ingestion of microplastics can impair nutrition of marine invertebrates. In a laboratory study, we tested whether microplastics affect ingestion rates and gastrointestinal enzyme activities in the marine isopod Idotea emarginata. Isopods were fed for eight days with one out of four different food formulations: natural food (the brown alga Fucus vesiculosus) or synthetic diet consisting of freeze-dried algal powder embedded in agarose, both, with or without microplastic particles (fluorescent polymethyl methacrylate, 10–100 μm) at a concentration of 40 items per mg of food. The isopods accepted both types of food but consumed significantly more (average 3.1-fold) of the agar based synthetic food. I. emarginata responded to the reduced content of digestible organic matter in the synthetic food by a compensatory adjustment of the ingestion rates. Addition of microplastics had no effect on ingestion rates in natural food whereas the feeding rates for synthetic food varied in response to microplastics. Similarly, activity patterns of digestive enzymes, particularly those of esterases, changed significantly in the treatment with synthetic food. Isopods fed with synthetic food alone showed elevated esterase activities in the gut while those isopods fed with synthetic food and microplastics showed elevated esterase activities in the midgut gland but not in the gut. Apparently, not the exposure to microplastic alone, but the combined effects of reduced nutrient availability and microplastic ingestion caused considerable biochemical reactions in the digestive organs of the isopods.

Lyso-DGTS lipid isolated from microalgae enhances PON1 activities in vitro and in vivo, increases PON1 penetration into macrophages and decreases cellular lipid accumulation.

High-density lipoprotein (HDL) plays an important role in preventing atherosclerosis. The antioxidant effect of HDL is mostly associated with paraoxonase 1 (PON1) activity. Increasing PON1 activity using nutrients might improve HDL function and quality and thus, decrease atherosclerotic risk. We previously isolated and identified a novel active compound, lyso-DGTS (C20:5,0) from Nannochloropsis sp. ethanol extract. In the present study, its effect on PON1 activities was examined and the mechanism by which the compound affects PON1 activity was explored. Lyso-DGTS elevated recombinant PON1 (rePON1) lactonase and esterase activities in a dose- and time-responsive manner, and further stabilized and preserved rePON1 lactonase activity. Incubation of lyso-DGTS with human serum for 4 h at 37 °C also increased PON1 lactonase activity in a dose-responsive manner. Using tryptophan-fluorescence-quenching assay, lyso-DGTS was found to interact with rePON1 spontaneously with negative free energy (ΔG = -22.87 kJmol-1 at 25 °C). Thermodynamic parameters and molecular modeling calculations showed that the main interaction of lyso-DGTS with the enzyme is through a hydrogen bond with supporting van der Waals interactions. Furthermore, lyso-DGTS significantly increased rePON1 influx into macrophages and prevented lipid accumulation in macrophages stimulated with oxidized low-density lipid dose-dependently. In vivo supplementation of lyso-DGTS to the circulation of mice fed a high-fat diet via osmotic mini-pumps implanted subcutaneously significantly increased serum PON1 lactonase activity and decreased serum glucose concentrations to the level of mice fed a normal diet. Our findings suggest a beneficial effect of lyso-DGTS on increasing PON1 activity and thus, improving HDL quality and atherosclerotic risk factors. © 2018 BioFactors, 44(3):299-310, 2018.

Quantification of GST and esterase activities in pyrethrin-resistant mosquitoes using pyrethroid-like fluorescent substrates

ABSTRACTBotanical pyrethrins and synthetic pyrethroids are highly potent and environmentally safe insecticides that are used to control a wide range of disease vector and pest arthropods. Unfortunately, resistance to these insecticides has been demonstrated in numerous medically important mosquito species. In this study, adult Culex pipiens sensu lato were captured in agricultural and urban locations in Fresno County, California, and subsequently exposed to a commercial formulation of pyrethrin insecticide by ultra-low-volume spraying. Following insecticide exposure, two pyrethroid-like, fluorescent substrates (4-methyl-2-oxo-2H-chromen-6-yl, cis-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropanecarboxylate (cis-DCVC) and 4-methyl-2-oxo-2H-chromen-6-yl, cis-3-((Z)-2-chloro-3,3,3-trifluoroprop-1-enyl)-2,2-dimethylcyclopropanecarboxylate (cis-TFMCVC)) and 1-chloro-2,4-dinitrobenzene (CDNB) were used to measure esterase and glutathione S-transferase (GST) activities in surviving mosquitoes. Elevated esterase activity (2.5-fold) was found in surviving urban mosquitoes at 12-h post-pyrethrin exposure (in comparison to non-insecticide-exposed control mosquitoes) when cis-TFMCVC was used as a substrate. Additionally, when CDNB was used as a substrate, 2.8-fold higher GST activity was found. A simple assay was established using our pyrethroid-like, fluorescent substrates that was able to detect low-level esterase activities in homogenates made from individual mosquitoes. The cis-TFMCVC-based assay suggested that esterase activity plays a role in pyrethrin resistance in urban mosquitoes in California.

Expression of fungal acetyl xylan esterase in Arabidopsis thaliana improves saccharification of stem lignocellulose.

Cell wall hemicelluloses and pectins are O-acetylated at specific positions, but the significance of these substitutions is poorly understood. Using a transgenic approach, we investigated how reducing the extent of O-acetylation in xylan affects cell wall chemistry, plant performance and the recalcitrance of lignocellulose to saccharification. The Aspergillus niger acetyl xylan esterase AnAXE1 was expressed in Arabidopsis under the control of either the constitutively expressed 35S CAMV promoter or a woody-tissue-specific GT43B aspen promoter, and the protein was targeted to the apoplast by its native signal peptide, resulting in elevated acetyl esterase activity in soluble and wall-bound protein extracts and reduced xylan acetylation. No significant alterations in cell wall composition were observed in the transgenic lines, but their xylans were more easily digested by a β-1,4-endoxylanase, and more readily extracted by hot water, acids or alkali. Enzymatic saccharification of lignocellulose after hot water and alkali pretreatments produced up to 20% more reducing sugars in several lines. Fermentation by Trametes versicolor of tissue hydrolysates from the line with a 30% reduction in acetyl content yielded ~70% more ethanol compared with wild type. Plants expressing 35S:AnAXE1 and pGT43B:AnAXE1 developed normally and showed increased resistance to the biotrophic pathogen Hyaloperonospora arabidopsidis, probably due to constitutive activation of defence pathways. However, unintended changes in xyloglucan and pectin acetylation were only observed in 35S:AnAXE1-expressing plants. This study demonstrates that postsynthetic xylan deacetylation in woody tissues is a promising strategy for optimizing lignocellulosic biomass for biofuel production.

Genetic Characterization of Esterase Activity Variants Associated with an Esterase Gene Amplification in a Strain of Culex pipiens from California.

In the Culex pipiens complex, a common mechanism of insecticide resistance is amplification of esterase genes leading to overproduction of detoxifying esterase enzymes. A number of electrophoretic esterase alleles have been identified, and in field populations individuals with the same esterase electromorph can exhibit a wide range of esterase enzyme activities. We isolated and characterized esterase activity variants associated with the esterase B1 electromorph from a field strain. A mating scheme was used to isolate chromosomes with esterase genes from the strain into 45 families. Twenty-six of the families received esterase genes from the field strain that conferred elevated esterase activity. Mean esterase activities in these families ranged from 43 to 695nmoles α-naphthyl acetate hydrolyzed/min/mg protein. Variance components indicated that genetic variance (i.e., genetic differences among families) accounted for 77% of the total variation in esterase activity. A comparison of mean esterase activities indicated that there were at least 11 different esterase activity variants contributing to the observed genetic variation in esterase activity among the 26 families. The relevance of these results to understanding the dynamics of amplified esterase genes in populations is discussed.

GENDER PECULIARITIES OF EXTERASE BLOOD NEUTROPHILE ACTIVITY IN PATIENTS WITH CHRONIC VIRAL HEPATITIS C

Aim. To study the changes in functional activity of blood neutrophiles in dynamics of treatment among patients with chronic viral hepatitis C (CVHC) depending on gender peculiarities. Materials and methods. 361 patients with CVHC were examined and treated. The disease had different degrees of activity. Exterase activity of neutrophiles was determined in dynamics – prior to and after treatment. The study was performed using R.P. Nartsissov’s technique. The results were calculated with Kaplow’s semiquantitative method. Results. Among CVHC patients with minimum degree of activity prior to treatment, men and women had tension of alphanaphthylbutyratesterase (BE) activity but alphanaphthylacetatesterase (AE) activity remained within the norm. After the basic therapy, BE activity was significantly inhibited, AE was intact. Against the background of complex therapy, men had inhibition of activity of both esterases. Women had an inhibited BE activity and elevated AE activity. Among CVHC patients with low degree of activity, prior to treatment esterase activity was decreased in men and elevated in women (AE), but BE – on the contrary, increased in men and decreased in women. After the basic therapy, men had an elevated esterase activity, but women – normalization of both esterases. After complex therapy, esterase activity completely normalized in both men and women. Application of basic and complex therapy leads to full normalization of esterases in women and AE in men. Conclusion. In CVHC patients with different degrees of activity there are differences in esterase activity of neutrophiles depending on gender peculiarities.

The pyrethroid resistance status and mechanisms in Aedes aegypti from the Guerrero state, Mexico

Dengue is one of the most important vector-borne diseases worldwide and is a public health problem in Mexico. Most programs in dengue endemic countries rely on insecticides for Aedes control. In Mexico, pyrethroid insecticides (mainly permethrin and deltamethrin) have been extensively used over a decade as adulticides and represented a strong selection for insecticide resistance for dengue vectors in several parts of the country. We studied the type, frequency and distribution of insecticide resistance mechanisms in Aedes aegypti from six municipalities in the state of Guerrero selected on the basis of historically intense chemical control and a high risk for dengue transmission. Ae. aegypti eggs were collected from October 2009 to January 2010 using ovitraps. F1 adults, emerged from these collections, were exposed to permethrin, deltamethrin and DDT in WHO diagnostic tests and showed high resistance levels to both pyrethroids and DDT. This was consistent with the presence of increased metabolic enzyme activities and target site insensitivity due to kdr mutations. Biochemical assays showed elevated esterase and glutathione S-transferase activities in the six municipalities. The V1016I kdr mutation on the IIS6 domain of the sodium channel gene was present in an overall frequency of 0.80. A second mutation, F1534C on the IIIS6 domain of the same gene was also detected, being the first report of this mutation in Guerrero. The multiple resistance mechanisms present in Ae. aegypti from Guerrero state represent a warning for the efficacy of the pyrethroid usage and consequently for the success of the dengue control program.

Variation of acephate susceptibility and correlation with esterase and glutathione S-transferase activities in field populations of the tarnished plant bug, Lygus lineolaris

The tarnished plant bug (TPB) has increasingly become an economically important pest of cotton. Heavy dependence on insecticides, particularly organophosphates and pyrethroids, for TPB control facilitated resistance development to multiple classes of insecticides. To better understand resistance and explore ways to monitor resistance in field populations, this study examined acephate susceptibility and the activities of two major detoxification enzymes in nine field populations collected in the Delta region of Mississippi and Arkansas in 2010. Two Arkansas populations from Reed and Backgate had 3.5- and 4.3-fold resistance to acephate, as compared to a susceptible laboratory strain. Extensive planting of cotton and heavy chemical sprays is a major driving force for resistance development to acephate in Mid-south cotton growing areas. Reduced susceptibility to acephate was highly correlated with elevated esterase activities. The acephate-resistant populations from Backgate, Lula, and Reed consistently had higher (up to 5.3-fold) esterase activities than susceptible populations. Regression analysis of LC50s with kinetic esterase activities revealed a significant polynomial quadratic relationship with R2 up to 0.89. Glutathione S-transferase (GST) also had elevated activity in most populations, but the variations of GST activities were not significantly correlated with changes of acephate susceptibility. Finally, examination of esterase and GST inhibitors indicated that suppression rates (up to 70%) by two esterase inhibitors in 2010 were slightly lower than those detected in 2006, and ethacrynic acid (EA) inhibited GST effectively in both years. Two other GST inhibitors (sulfobromophthalein and diethyl maleate) displayed significantly lower suppression rates in 2010 than those detected in 2006, suggesting a potential genetic shift in pest populations and a necessity of continued monitoring for insecticide resistance with both bioassay and biochemical approaches. Results indicated that using major detoxification enzyme activities for resistance monitoring may provide insight into acephate resistance in field populations of TPB.

Comparative evaluation of the protective potentials of human paraoxonase 1 and 3 against CCl 4-induced liver injury

We previously reported that electroporation mediated hPON1 or hPON3 gene delivery could protect against CCl 4-induced liver injury. However, substantial evidence supported that the in vivo physiological functions of hPON1 and hPON3 were distinct. To compare the protective efficacies of hPON1 and hPON3 against liver injury, recombinant adenovirus AdPON1 and AdPON3, which were capable of expressing hPON1 and hPON3 respectively, were intravenously injected into mice before they were given CCl 4. Adenovirus mediated expression of hPON1 and hPON3 were demonstrated by elevated serum esterase activity, hepatic lactonase activity, and hPON1/hPON3 mRNA expression in liver. Serum transaminase assay, histological observation and TUNEL analysis revealed that the extent of liver injury and hepatocyte apoptosis in AdPON1 or AdPON3 treated mice was significantly ameliorated in comparison with control. Meanwhile, overexpression of hPON1 and hPON3 reduced the hepatic oxidative stress and strengthen the total antioxidant capabilities in liver through affecting the hepatic malondialdehyde (MDA), glutathione (GSH) and total antioxidant capability (T-AOC) levels, regardless of the exposure to CCl 4 or corn oil. Administration of AdPON1 or AdPON3 also suppressed inflammatory response by decreasing TNF-α and IL-1β levels in CCl 4 mice. In this study, hPON1 exhibited a slightly higher efficacy than hPON3 in alleviating liver injury, but the difference between them were not significant.

Esterase-based metabolic resistance to insecticides in heliothine and spodopteran pests

Elevated esterase activities and increased band intensities of multiple esterase isozymes after electrophoresis are commonly associated with resistance to organophosphate, pyrethroid and carbamate insecticides in various heliothine and spodopteran pests. One possible explanation for this involves a ‘master regulator’ mutation in a more general chemical stress response. An association between elevated esterase activities and isozyme intensities has also been reported for resistance to the Cry1Ac toxin of Helicoverpa armigera. The basis for this is unclear albeit some involvement of esterases could be mediated by the toxin's affinity for N-acetyl galactosamine glycans on certain gut-expressed esterases in this species.

Pyrethroid resistance and esterase activity in three strains of the cotton bollworm, Helicoverpa armigera (Hübner)

Microplate assay technique for estimation of esterase activity in a single insect was used in combination with dose mortality bioassays to detect pyrethroid resistance in three strains of Helicoverpa armigera and to study the frequency of pyrethroid resistant individuals within the population of the same strain at two larval stages, third and fifth instar. The third and fifth instar larvae of the field strains i.e., Nagpur strain and Delhi strain that displayed high degree of resistance towards deltamethrin, had higher esterase activity compared to a susceptible laboratory strain. The frequency distribution of individuals with elevated esterase activity above 1.00 absorbance unit was correlated with the resistance level of the strains. The frequency of resistant individuals in the third instar larvae of Nagpur strain and Delhi strain were 29% and 23%, respectively compared to 4% in the susceptible strain. The resistant individuals in the resistant strains have markedly increased in the fifth instar larvae with a frequency distribution of 63% and 90% in Delhi strain and Nagpur strain, respectively, while only 14% of individuals was found to have elevated esterase activity in the susceptible strain. The results demonstrated the role of esterase in pyrethroid resistance in H. armigera. Microplate assay proved to be a rapid and reliable biochemical technique for monitoring of pyrethroid resistance in H. armigera.

Susceptibility of <I>Culex pipiens</I> (Diptera: Culicidae) Field Populations in Cyprus to Conventional Organic Insecticides, <I>Bacillus thuringiensis</I> subsp. <I>israelensis</I>, and Methoprene

Culex pipiens pipiens L. populations on Cyprus were sampled over a 6-yr period from 2002 to 2008 to evaluate the status of insecticide resistance toward the insecticides temephos, chlorpyrifos, and permethrin and to study susceptibility levels toward the recently introduced bacterial insecticide Bacillus thuringiensis subsp. israelensis De Barjac and the juvenile hormone analog, methoprene. Susceptibility to the three conventional chemical insecticides varied between different collections, with most collections showing moderate or low resistance. The 2004 Akrotiri collection had the highest temephos resistance ratio, 167-fold at the LC95, although later sampling showed that the population returned to susceptibility after treatments stopped. Chlorpyrifos resistance was generally higher than temephos resistance. Four collections showed high resistance, and the resistance ratios of two collections were notably high with resistance ratios of 110- and 248-fold at the LC95. Three collections showed high permethrin resistance (22.5-, 23.9-, and 86.3-fold). The frequency of elevated esterase activity in populations was estimated using a filter paper test, and frequencies varied from 0.9 to 65% among collections. The levels of temephos resistance and the frequency of elevated esterases in this survey were generally lower than in earlier reports, suggesting a decline in temephos resistance. Dose-response values for B. thuringiensis subsp. israelensis covered an approximate eight-fold range, but no resistance was detected. Methoprene values showed a 4.7-fold and 16-fold range at the LC50 and LC95, respectively. Two populations showed significant resistance ratios at the LC95. These data are discussed in relation to the changes in larval control practices underway in Cyprus.

Insecticide resistance and detoxifying enzyme activity in the principal bancroftian filariasis vector, Culex quinquefasciatus, in northeastern India

The insecticide resistance status of Culex quinquefasciatus Say (Diptera: Culicidae) to DDT and deltamethrin across army cantonments and neighbouring villages in northeastern India was investigated. In India, DDT is still the insecticide of choice for public health programmes. In military stations, pyrethroids, especially deltamethrins, are used for insecticide-treated nets (ITNs). Recent information on the levels of resistance to DDT and deltamethrin in mosquito populations of northeastern India is scare. Continued monitoring of insecticide resistance status, identification of the underlying mechanisms of resistance in local mosquito populations and the establishment of a baseline data bank of this information are of prime importance. Insecticide susceptibility assays were performed on wild-caught adult female Cx. quinquefasciatus mosquitoes to the discriminating doses recommended by the World Health Organisation (WHO) to DDT (4%) and deltamethrin (0.05%). Across all study sites, mortality as a result of DDT varied from 11.9 to 50.0%, as compared with 91.2% in the susceptible laboratory strain (S-Lab), indicating that Cx. quinquefasciatus is resistant to DDT. The species was found to be 100% susceptible to deltamethrin in all study sites except Benganajuli and Rikamari. Knock-down times (KDT) in response to deltamethrin varied significantly between study sites (P < 0.01) from 8.3 to 17.8 min for KDT(50) and 37.4 to 69.5 min for KDT(90). All populations exceeded the threshold level of alpha-esterase, beta-esterase and glutathion S-transferase (GST) established for the S-Lab susceptible strain, and all populations had 100% elevated esterase and GST activity, except Missamari and Solmara. Beta-esterase activity in Field Unit II (96.9%) was less than in any of the other populations. Benganajuli had the highest activity level for all the enzymes tested. There was a significant correlation between all enzyme activity levels and insecticide resistance phenotype by populations (P < 0.05). The results presented here provide the first report and baseline information of the insecticide resistance status of Cx. quinquefasciatus in northeastern India, and associated information about biochemical mechanisms that are essential for monitoring the development of insecticide resistance in the area.

The Susceptibilities of Oxya chinensis (Orthoptera: Acridoidea) to Malathion and Comparison of the Esterase Properties from Three Collected Populations in Tianjin Area, China

The susceptibilities of Oxya chinensis to malathion were studied in three populations collected from outskirt of Tianjin, China, using bioassays and biochemical analysis. Populations were chosen to represent different exposure to insecticides: BDG (Beidagang; low exposure), BD (Baodi; high exposure previously but low exposure now), and JN (Jinnan; high exposure). The results showed that the LD 50 values of BD and JN populations were 3.95- and 12.02-fold and 3.64- and 10.07- fold higher than that of BDG population in females and males, respectively. The LD 50 values in females were higher than those in males. The results of biochemical analysis indicated that the esterase (EST) activities in JN population were higher than those in BD and BDG populations. They showed that when α-NA, α-NB, and β-NA were used as substrates, females' EST activities of JN population were 1.11-, 1.30-, and 1.14-fold and 1.39-, 1.59-, and 1.54-fold higher than those of BD and BDG populations, respectively. When α-NA, α-NB, and β-NA were used as substrates, males' EST activities of JN population were 1.13-, 1.12-, and 1.00-fold and 1.20-, 1.14-, and 1.07-fold higher than those of BD and BDG populations, respectively. The results also showed that the specific activities of the females were higher than those of the males in the BD and JN populations, whereas the specific activities of the males were higher than those of the females in the BDG population. The results of bioassay were consistent with those of biochemical analysis. Thus, it was inferred that the elevated ESTs activities might play an important role in conferring the differences of susceptibility of O. chinensis to malathion in the three collected populations. Enzyme kinetic studies indicated that the K m and V max values were different among the three collected populations and between the females and the males. The observed changes in the kinetic parameters might be explained by differential expression patterns of isozymes so that the insect esterases have different affinities and maximum velocities toward the same substrate.

Enzymatic activities ofCandida tropicalisisolated from hospitalized patients

Secretion of hydrolytic enzymes is considered a virulence factor in Candida spp. Extracellular enzymatic activities in 29 clinical isolates of Candida tropicalis were analyzed by plate assays. C. tropicalis, similar to Candida albicans, showed elevated hemolytic and esterase activities. However, unlike C. albicans, low aspartyl protease and very low phospholipase activities were detected in C. tropicalis isolates.