Improved synthesis of the antifungal isobutyl o-coumarate catalyzed by the Aspergillus terreus type B feruloyl esterase

BACKGROUND Hydroxycinnamic acids and some of their derivatives are molecules with interesting biological activities; for instance, hydroxylated hydroxycinnamic esters have proved to have antifungal properties, and thus the generation of these molecules is of industrial importance. In this study, the direct esterification capacity of the pure recombinant type B feruloyl esterase from Aspergillus terreus (AtFAE B) was evaluated by its ability to catalyze the synthesis of isobutyl o-coumarate, an interesting antifungal molecule. A ternary solvent system (isooctane/isobutanol/water) was employed to improve the synthesis of isobutyl o-coumarate, assessing different substrate concentrations, enzyme load, water percentages and pH and temperature values. RESULTS AtFAE B showed the highest initial rate at 18% (v/v) isobutanol and 50 mM o-coumaric acid, 0.04 mg/ml of enzyme, 4% (v/v) water without buffer and 40C. AtFAE B half-lives at 30C, 40C and 50C were 16.5 h, 1.75 h and 3.5 min, respectively. Thus, we decided to evaluate the bioconversion yield at 30C, where the enzyme showed the highest operational stability. At this temperature, we obtained a yield of ~80% after only 8 h of reaction, using a 78:18:4 isooctane:isobutanol:water ternary solvent system, with 50 mM of o-coumaric acid.CONCLUSIONS Under these improved conditions, the productivity was 1.06 g isobutyl o-coumarate/L*h with a biocatalyst yield of 209.6 kg isobutyl o-coumarate/kg free AtFAE B, demonstrating the promising potential of AtFAE B to accept the non-canonical o-coumaric acid as the substrate and to achieve the synthesis of isobutyl o-coum

Caracterización, clasificación y usos de las enzimas lipasas en la producción industrial / Characterization, classification and uses of lipase enzymes in industrial production

Introducción: La bioquímica, como ciencia particular dentro de las ciencias médicas, ha tenido un gran desarrollo. Las enzimas lipasas se obtienen de organismos vivos que abundan en la naturaleza y han sido utilizadas en la producción de alimentos, jabones, detergentes, aceites y otros productos industriales. Actualmente se han logrado nuevas clasificaciones de estas, subdivididas en grupos y subgrupos. Se aprecia además interés de utilizarlas en la producción de biodiesel y en la biotecnología y genética médica. Objetivo: Recopilar las principales consideraciones teóricas actualizadas acerca la caracterización, clasificación y usos de las enzimas lipasas. Método: La búsqueda y análisis de la información se realizó desde el primero de septiembre al 23 de diciembre de 2019, con un total de 50 artículos publicados en las bases de datos PubMed, Hinari, SciELO y Medline, mediante el gestor de búsqueda y administrador de referencias EndNote. se utilizaron 42 citas seleccionadas para realizar la revisión, de ellas 38 de los últimos cinco años. Conclusiones: Las enzimas lipasas son proteínas que catalizan procesos biológicos. son activas en un amplio rango de sustrato, realizan reacciones de síntesis, hidrólisis o de intercambio de grupos. Poseen diversas actividades catalíticas, son menos costosas y menos contaminantes, se obtienen en gran cantidad, se producen de forma regular. Son estables y su proceso de producción es más factible y seguro. Se caracterizan por su capacidad de catalizar reacciones de acidólisis, alcohólisis, aminólisis, esterificación, interesterificación y transesterificación, entre otras características(AU)

Introduction: Biochemistry has experienced great development as a particular medical science. Lipase enzymes are obtained from living organisms which are abundant in nature, and have been used in the manufacture of foods, soap, detergents, oils and other industrial products. New classifications are now available of lipase enzymes, and they have been subdivided into groups and subgroups. An interest is also noticed in using them for biodiesel production and in biotechnology and medical genetics. Objective: Collect the main updated theoretical considerations about the characterization, classification and uses of lipase enzymes. Method: The search for and analysis of the information extended from 1 September to 23 December 2019, for a total 50 papers published in the databases PubMed, Hinari, SciELO and Medline, using the search engine and reference manager EndNote. Forty-two citations were selected for the review, 38 of which were from the last five years. Conclusions: Lipase enzymes are proteins that catalyze biological processes. They are active in a wide range of substrates, performing synthesis reactions, hydrolysis or group exchanges. They display a variety of catalytic activities, are less costly and less contaminating, are obtained in large quantities and are produced in a regular manner. They are stable and their production process is more feasible and safer. They are characterized by their ability to catalyze reactions of acidolysis, alcoholysis, aminolysis, esterification, interesterification and transesterification, among other characteristics(AU)

Effects of biopesticides in Tetragonisca angustula latreille (Hymenoptera: Meliponinae) pollinators / Efeitos de biopesticidas em polinizadores Tetragonisca angustula latreille (Hymenoptera: Meliponinae) / Efectos de los bioplaguicidas sobre los polinizadores Tetragonisca angustula latreille (Hymenoptera: Meliponinae)

Stingless bees Tetragonisca angustula (Latreille) (Hymenoptera: Meliponinae) are pollinators of native and cultivated plants and are therefore in contact with areas contaminated by pesticides. These native bees were evaluated for changes in gene expression of esterase isoenzymes (EST) and peptides after contamination by contact with growth regulators from insecticides Gallaxy® EC 100, Natuneem and Azamax after 48, 120, 168 hours, 30 and 60 days. EST-4 presented an increase in relative activity after contamination with Gallaxy® EC 100 at 6.2 × 10-2 g a.i./mL; Natuneem at 7.5 × 10-5 g a.i./mL; and Azamax at 1.2 × 10-3 g a.i/mL after 60 days, 48 h, and 60 days, respectively. Inhibition of the relative activity of EST-4 was detected after contamination by Natuneem at 1.5 × 10-5 g a.i./mL and Azamax at 1.2 × 10-3 g a.i./mL after 48 h and 30 days, respectively. The insecticide growth regulators promoted changes in protein synthesis of T. angustula adult workers resulting in an increase or decrease in the relative intensity of bands, and the appearance of new peptides when compared with controls. Changes in protein synthesis have been identified mainly after long period of contamination, 120 and 168 h with the IGRs Gallaxy® EC 100 (at 0.78 and 1.25 g a.i./mL), Azamax (at 1.2 × 10-3 and 6 × 10-3 g a.i./mL), and Natuneem (at 7.5 × 10-5 and 3 × 10-3 g a.i./mL), and at 60 days with Natuneem (at 1.5 × 10-5 g a.i./mL).(AU)

Abelhas sem ferrão Tetragonisca angustula (Latreille) (Hymenoptera: Meliponinae) são polinizadores de plantas nativas e cultivadas e, portanto, estão em contato com áreas contaminadas por biopesticidas. Essas abelhas nativas foram avaliadas quanto a alterações na expressão gênica de isoenzimas esterases (EST) e peptídeos após contaminação por contato com reguladores de crescimento de inseticidas Gallaxy® EC 100, Natuneem e Azamax após 48, 120, 168 horas, 30 e 60 dias. A EST-4 apresentou um aumento na atividade relativa após a contaminação com Gallaxy® 100 EC em 6,2 × 10-2 g i.a./mL, Natuneem em 7,5 × 10-5 g i.a./mL e Azamax em 1,2 × 10-3 g i.a./mL após 60 dias, 48 h e 60 dias, respectivamente. A inibição da atividade relativa de EST-4 foi detectada após contaminação pelo Natuneem a 1,5 × 10-5 g i.a./mL e Azamax a 1,2 × 10-3 g i.a./mL após 48 he 30 dias, respectivamente. Os reguladores de crescimento de inseticidas promoveram alterações na síntese protéica de trabalhadores adultos de T. angustula, resultando em um aumento ou diminuição da intensidade relativa das bandas e no aparecimento de novos peptídeos em comparação com os controles. Alterações na síntese de proteínas foram identificadas principalmente após um longo período de contaminação, 120 e 168 h com o IGRs Gallaxy® EC 100 (0,78 e 1,25 g i.a./mL), Azamax (1,2 × 10-3 e 6 × 10-3 g i.a./mL) e Natuneem (7,5 × 10-5 e 3 × 10-3 g i.a./mL) e 60 dias com Natuneem (1,5 × 10-5 g i.a./mL).(AU)

Las abejas sin aguijón Tetragonisca angustula (Latreille) (Hymenoptera: Meliponinae) son polinizadores de plantas nativas y cultivadas y, por lo tanto, están en contacto con áreas contaminadas por bioplaguicidas. Estas abejas nativas fueron evaluadas para detectar cambios en la expresión génica de isoenzimas esterasa (EST) y péptidos después de la contaminación por contacto con los reguladores del crecimiento insecticidas Gallaxy® EC 100, Natuneem y Azamax después de 48, 120, 168 horas, 30 y 60 días. EST-4 mostró un aumento en la actividad relativa después de la contaminación con Gallaxy® 100 EC a 6.2 × 10-2 g i.a./mL, Natuneem a 7.5 × 10-5 g i.a./mL y Azamax a 1.2 × 10-3 g i.a./mL después de 60 días, 48 hy 60 días, respectivamente. La inhibición de la actividad relativa de EST-4 se detectó después de la contaminación por Natuneem a 1.5 × 10-5 g i.a./mL y Azamax a 1.2 × 10-3 g i.a./mL después de 48 hy 30 días. respectivamente. Los insecticidas reguladores del crecimiento promovieron cambios en la síntesis de proteínas de trabajadores adultos de T. angustula, resultando en un aumento o disminución de la intensidad relativa de las bandas y en la aparición de nuevos péptidos en relación a los controles. Los cambios en la síntesis de proteínas se identificaron principalmente después de un largo período de contaminación, 120 y 168 h con IGRs Gallaxy® EC 100 (0.78 y 1.25 g i.a./mL), Azamax (1.2 × 10-3 y 6 × 10-3 g i.a./mL) y Natuneem (7.5 × 10-5 y 3 × 10-3 g i.a./mL) y 60 días con Natuneem (1.5 × 10-5 g i.a./mL).(AU)

Determination of insecticides' lethal concentrations and metabolic enzyme levels in Triatoma dimidiata / Determinación de concentraciones letales y niveles de enzimas metabólicas de insecticidas en Triatoma dimidiata

Abstract: Objective: The feasibility of the use of WHO impregnated paper and biochemical assays to determine lethal concentrations (LC50 and LC99) and insecticide metabolic enzyme levels of Triatoma dimidiata. Materials and methods: LC50 and LC99 were calculated with WHO papers impregnated at different concentrations of malathion, propoxur and deltamethrin; the percentage of insensitive acetylcholinesterase (iAChE); and the levels of esterases, glutathione S-transferases, and monooxygenases in laboratory nymphs of the first stage (5 to 7 days), were undertaken using the WHO biochemical assays. Results: Respectively the LC50 and LC99 µg/cm2 obtained for malathion were 43.83 and 114.38, propoxur 4.71 and 19.29, and deltamethrin 5.80 and 40.46. A 30% of the population had an iAChE, and only a few individuals had high P450 and β-eterase levels. Conclusion: Impregnated papers and biochemical tests developed by WHO for other insects, proved to be feasible methods in monitoring insecticide resistance and metabolic enzymes involved in T. dimidiata.

Resumen: Objetivo: La factibilidad de usar los papeles impregnados y ensayos bioquímicos según la OMS para determinar concentraciones letales (CL50 y CL99) y niveles enzimáticos en la resistencia a insecticidas en Triatoma dimidiata. Material y métodos: Se calcularon la CL50 y CL99 con papeles impregnados según la OMS a diferentes concentraciones de malatión, propoxur y deltametrina; el porcentaje de acetilcolinesterasa insensible (iAChE); y los niveles de esterasas, glutatión S-transferasas, y monooxigenasas en ninfas de laboratorio del estadio I (5-7 días) se determinaron usando los ensayos bioquímicos según la OMS. Resultados: Se obtuvieron las CL50 y CL99 µg / cm2 respectivamente para malatión 43.83 y 114.38, propoxur 4.71 y 19.29, y deltametrina 5.80 y 40.46. Un 30% de las chinches tuvo iAChE, y sólo pocos individuos tuvieron niveles superiores de P450 y β-eterasas. Conclusión: Los papeles impregnados y ensayos bioquímicos que describe la OMS para otros insectos demostraron ser métodos factibles para monitorear la resistencia a insecticidas y las enzimas metabólicas involucradas en T. dimidiata.

Bioprospection of proteases from Halobacillus andaensis for bioactive peptide production from fish muscle protein

BACKGROUND: Biologically active peptides produced from fish wastes are gaining attention because their health benefits. Proteases produced by halophilic microorganisms are considered as a source of active enzymes in high salt systems like fish residues. Hence, the aim of this study was the bioprospection of halophilic microorganisms for the production of proteases to prove their application for peptide production. RESULTS: Halophilic microorganisms were isolated from saline soils of Mexico and Bolivia. An enzymatic screening was carried out for the detection of lipases, esterases, pHB depolymerases, chitinases, and proteases. Most of the strains were able to produce lipases, esterases, and proteases, and larger hydrolysis halos were detected for protease activity. Halobacillus andaensis was selected to be studied for proteolytic activity production; the microorganism was able to grow on gelatin, yeast extract, skim milk, casein, peptone, fish muscle (Cyprinus carpio), and soy flour as protein sources, and among these sources, fish muscle protein was the best inducer of proteolytic activity, achieving a protease production of 571 U/mL. The extracellular protease was active at 50°C, pH 8, and 1.4 M NaCl and was inhibited by phenylmethylsulfonyl fluoride. The proteolytic activity of H. andaensis was used to hydrolyze fish muscle protein for peptide production. The peptides obtained showed a MW of 5.3 kDa and a radical scavenging ability of 10 to 30% on 2,2-diphenyl-1-picrylhydrazyl and 2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) and a ferric reducing ability of plasma. Conclusion: The use of noncommercial extracellular protease produced by H. andaensis for biologically active peptide production using fish muscle as the protein source presents a great opportunity for high-value peptide production.

Estudio preliminar sobre el comportamiento de Aedes albopictus de la region central de Venezuela a insecticidas químicos / Preliminary study on the behavior of Aedes albopictus from the central region of Venezuela to chemical insecticides

Aedes albopictus es un competente vector del virus chikungunya, zika y de la mayor parte de los virus de encefalitis equina, capaz de transmitir al menos 24 arbovirus, susceptible a la infeccion (vertical y horizontalmente) por los serotipos del virus dengue. El objetivo fue realizar un estudio preliminar sobre el comportamiento frente a insecticidas organosinteticos de Ae. albopictus de diferentes localidades de tres estados centrales del pais, durante el periodo 2012-2014. Se realizaron pruebas biologicas con botellas tratadas con insecticidas, segun el metodo del CDC e igualmente evaluacion bioquimica para identificar los mecanismos de detoxificacion enzimaticos in vitro y mecanismos de resistencia in vivo con el sinergista butoxido de piperonilo (PB). Todas las cepas evaluadas presentaron sensibilidad al DDT (200 µg/mL) y cuatro de ellas a lambdacialotrina (6,25 µg/mL), solo una, Santos Michelena (SM) presento valores de sobrevivencia. Tambien esto fue observado con: malation (100 µg/mL) con una mortalidad en 30 minutos de 42,6% para Carabobo; 46,2% Distrito Capital (DC); 69,6% Zamora y 75,5% Mario Briceno Iragorry (MBI), solo resulto sensible SM. En cuanto a fenitrotion se encontraron valores de sobrevivencia con mortalidades en 30 minutos de 72,2%; para Carabobo, 30,7% DC y 45,8% MBI; solo expresaron sensibilidad Zamora y SM. En las cepas Carabobo y DC se observo el efecto sinergico FS = 3 y FS = 1,3, respectivamente. Se presume que los valores de sobrevivencia encontrados para insecticidas organofosforados pudiera asociarse con el incremento de esterasas alfa (a) y beta (b), en menor medida por la acetilcolinesterasa insensible (AChei) y las oxidasas con la sobrevivencia a lambdacialotrina en la cepa SM, lo cual podria comprobarse en estudios futuros a realizarse sobre resistencia a insecticidas. Los resultados encontrados en el presente trabajo, constituyen la base para su inicio a corto plazo en Ae. albopictus de Venezuela y la region, a fin de contribuir en la eficacia de las medidas de control quimico de esta especie de importancia medica.

Aedes albopictus is a competent vector of chikungunya virus, zika and most of the equine virus, capable of transmitting at least 24 arboviruses, susceptible to infection (vertically and horizontally) by serotypes of the dengue virus encephalitis. The objective was to conduct a preliminary study on the behavior against Ae. albopictus organosintetic insecticides from different locations in three central states of the country during the period 2012-2014. Bioassays with insecticide-treated bottles were performed according to the method of the CDC and also biochemical evaluation to identify the mechanisms of detoxification enzyme in vitro and in vivo mechanisms of resistance to the synergist piperonyl butoxide (PB). All strains tested showed sensitivity to DDT (200 µg/mL) and four of them lambdacyhalothrin (6.25 µg/mL), only one, Santos Michelena (SM) presented values of survival. This was also observed: malathion (100 µg/mL) with a mortality time (30 min) of 42.6% for Carabobo; 46.2% Capital District (DC); Zamora 69.6% and 75.5% Mario Briceno Iragorry (MBI) was sensitive only SM. As fenitrothion survival values encountered in the mortalities time (30 min) 72.2%; for Carabobo, 30.7% and 45.8% MBI DC; only expressed sensitivity Zamora and SM. In DC strains Carabobo and the synergistic effect was observed FS = 3 and FS = 1.3, respectively.It is presumed that the survival values found for organophosphate insecticides may be associated with increased esterase alpha (a) and beta (b), to a lesser extent insensitive acetylcholinesterase (ACHEI) and oxidases with survival to lambdacialotrine in strain SM , which could be verified in future studies to be carried out on insecticides resistance. The results found in the present work, constitute the basis for its short-term start in Ae. albopictus from Venezuela and the region, in order to contribute to the effectiveness of chemical control measures of this species of medical importance.

Estado de la resistencia a insecticidas organosintéticos de Aedes aegypti de Coro, estado Falcon, Venezuela / Status of resistance to organo-synthetic insecticides by an Aedes aegypti strain from Coro, Falcon state, Venezuela

Aedes aegypti es una de las especies más estudiadas en salud pública por ser vector primario en el ser humano del virus dengue, fiebre amarilla, chikungunya y zika. Se evaluó la respuesta a los insecticidas organofosforados malatión, fenitrotión, pirimifos-metil al carbamato propoxur y al piretroide lambdacialotrina en una cepa de campo denominada Coro, conformada por un pool de material (inmaduro) colectado en barriadas con alta casuística de dengue ubicadas en municipio Miranda, estado Falcón, Venezuela; en comparación con la cepa susceptible Rockefeller. Se realizaron los bioensayos en botellas tratadas con insecticidas, según el método del CDC e igualmente se identificaron los mecanismos de resistencia in vitro a través de pruebas bioquímicas. La cepa evaluada resultó resistente a malatión (45 min), fenitrotión (75 min) pirimifos-metil (75 min) con respecto a la cepa susceptible con 30, 45 y 45 min.respectivamente, con porcentajes de mortalidad en el tiempo umbral de 89,2% malatión; 55,7% fenitrotión; 56,6%, pirimifos-metil. Se encontró susceptibilidad al carbamato propoxur y al piretroide lambdacialotrina. En cuanto a las pruebas bioquímicas se encontraron valores elevados para esterasas alfa (α) y beta (β) con diferencias estadísticamente significativas (P<0,05) al comparar con la cepa susceptible. La acetilcolinesterasa inhibida (Achei) a pesar de encontrar diferencias significativas con la cepa Rockefeller los valores están por debajo de la misma. La resistencia de la cepa Coro a los insecticidas organofosforados parece asociarse con el incremento de las esterasas alfa (α) y beta (β). Los resultados obtenidos representan un aporte valioso para monitorear la resistencia de Ae. aegypti a nivel local dentro de un programa de manejo integrado de vectores.

Aedes aegypti is one of the most studied species in public health as the primary vector in humans the dengue, chicunguya, zyca virus and yellow fever. Response to organophosphorus insecticides malathion, fenitrothion, methyl-pirimiphos and carbamate propoxur was evaluated and the pyrethroid lambda-field strain called Coro, made up of a pool of material (immature) collected in neighborhoods with high casuistry of dengue Coro located in Miranda municipality, Falcón state, Venezuela; compared with the susceptible Rockfeller. Bioassays in insecticide-treated bottles were performed according to the method of the CDC and also the mechanisms of resistance in vitro were identified by biochemical tests. The resistant strain was evaluated to malathion (45 min), fenitrothion (75 min) methyl-pirimiphos (75 min) compared to the susceptible strain with 30, 45 and 45 min respectively, with mortality rates of 89 time threshold, 2% malathion, fenitrothion 55.7%, 56.6%, pirimiphos-methyl. Susceptibility to propoxur carbamate and pyrethroid lambda was found. As for the high values for biochemical tests esterases alpha (α) and beta (β) with statistically significant differences (P< 0.05) compared with the susceptible strain were found. Inhibited aceltilcholinesterase (Achei) despite significant differences with the Rockefeller strain values are below the same acetylcholinesterase. Falcon resistance to organophosphate insecticides strain appears to be associated with increased alpha esterases (α) and beta (β). The results represent a valuable contribution to monitor the resistance of Ae. aegypti locally within a program of integrated vector management.

Esterase isozymes patterns of grape vine (Vitis vinifera L) are altered in response to fungicide exposure / Padrões de isozimas Esterases de videira (Vitis vinifera L) alterados em resposta à exposição a fungicidas

Current analysis characterizes the effect of different fungicides often applied for pest control on α−and ß-esterase patterns of four economically important table-wine grape cultivars (Italia, Rubi, Benitaka and Brasil) of Vitis vinifera. The α- and ß-esterase patterns in bud leaves of the cultivars were assessed by native PAGE analysis. Cabrio Top® compound inhibited EST-2, EST-5, EST-6, EST-7, EST-8, EST-9 and EST-10 carboxylesterases, whereas EST-4, EST-11, EST-12, EST-13, EST-14 acetylesterases and EST-16 carboxylesterase were detected as weakly stained bands. Carboxylesterases and acetylesterases were also detected as weakly stained bands when exposed to fungicides Orthocide 500®, Positron Duo® and Folicur PM®. No changes in α- and ß-esterase patterns were reported when the vines were exposed to the fungicides Rovral SC®, Kumulus DF®, Curzate M®, Score® or Cuprogarb 500®. The evidence of functional changes in carboxylesterase and acetylesterase levels in current study is a warning to grape producers on the dangers inherent in the indiscriminate use of potent and modern fungicides extensively used in agriculture. The inhibition effect of fungicides on esterase isozyme molecules seems to be independent of the fungicide chemical.

O presente estudo caracterizou o efeito de diferentes fungicidas comumente aplicados como medidas de controle de pragas sobre padrões de α- e ß-esterases de quatro importantes cultivares de uva de mesa (Itália, Rubi, Benitaka e Brasil) de Vitis vinifera. Os padrões de α- e ß-esterases de brotos foliares das cultivares foram avaliados por PAGE. O composto Cabrio Top® inibiu as carboxilesterases EST-2, EST-5, EST-6, EST-7, EST-8, EST-9 e EST-10, enquanto as acetilesterases EST-4, EST-11, EST-12, EST-13, EST-14 e a carboxilesterase EST-16 foram detectadas como bandas fracamente coradas. As carboxilesterases e acetilesterases também foram detectadas como bandas fracamente coradas quando expostas aos fungicidas Orthocide 500®, Positron Duo® e Folicur PM®. Não foram observadas alterações nos padrões de α- e ß-esterases quando as videiras foram expostas aos fungicidas Rovral SC®, Kumulus DF®, Curzate M®, Score® ou Cuprogarb 500®. A evidência de alterações em nível funcional em carboxilesterases e acetilesterases, apresentada neste estudo, pode servir como um alerta aos produtores de uva dos perigos inerentes ao uso indiscriminado de fungicidas potentes e modernos amplamente utilizados hoje na agricultura. O efeito dos fungicidas sobre as enzimas esterases parece ser independente do grupo químico ao qual pertence o fungicida.

EVALUATION OF LEUKOCYTE ESTERASE REAGENT STRIPS TEST IN THE DIAGNOSIS OF SPONTANEOUS BACTERIAL PERITONITIS IN CHILDREN WITH CIRRHOSIS / Avaliação de tiras reagentes de teste de esterase de leucócitos no diagnóstico de peritonite bacteriana espontânea em crianças com cirrose

BackgroundSpontaneous bacterial peritonitis is defined as an ascetic fluid infection without an evident intra-abdominal surgically treatable source. Spontaneous bacterial peritonitis is one of the severe complications in patients with cirrhosis and ascites. Without early antibiotic treatment, this complication is associated with high mortality rate; therefore, early diagnosis and treatment of spontaneous bacterial peritonitis is necessary for survival. Leukocyte esterase reagent can rapidly diagnose the spontaneous bacterial peritonitis.ObjectiveThis study aimed to find out the diagnostic accuracy of leukocyte esterase dipstick test for the diagnosis of spontaneous bacterial peritonitis.MethodsA single centered hospital-based cross-sectional study was conducted during July 2013 to August 2014 on children with cirrhotic liver disease and ascites who were admitted in the Department of Pediatric Gastroenterology in Nemazee Hospital affiliated to Shiraz University of Medical Sciences (Iran). All patients underwent abdominal paracentesis, and the ascitic fluid was processed for cell count, leukocyte esterase reagent strip test (Combiscreen SL10) and culture. Spontaneous bacterial peritonitis was defined as having a polymorphonuclear count (PMN ≥250/m3) in ascitic fluid. Sensitivity, specificity, positive predictive value and negative predictive value of leukocyte esterase test were calculated according to the formula.ResultsTotally, 150 ascitic fluid sample of cirrhotic male patients (53.2%) and their mean age (4.33±1.88 years) were analyzed. Biliary atresia (n=44, 29.4%) and idiopathic neonatal hepatitis (n=29, 19.3%) were the most frequent etiology of cirrhosis. Also, abdominal pain (68.6%) and distension (64%) were the most common presenting complaint. Of all cases, 41patients (27.35%) were diagnosed to have spontaneous bacterial peritonitis (PMN ≥250/mm3). Sensitivity and specificity of leukocyte esterase reagent test according to PMNs ≥250mm3 were 87.80% and 91.74%, also on ascitic fluid culture results were 88.23% and 77.44%. Positive predictive value and negative predictive value of this test in PMNs ≥250mm3 were 80% and 95.23% and in cases with positive culture 33.33% and 98.09% were obtained, respectively. Efficiency of leukocyte esterase reagent test in diagnosing spontaneous bacterial peritonitis, according to PMNs ≥250mm3 and culture results were 90.66% and 78.66%.ConclusionThe leukocyte esterase strip test may be used as rapid test for diagnosis of spontaneous bacterial peritonitis due to its high diagnostic validity.

ContextoA peritonite bacteriana espontânea é definida como uma infecção do fluido ascítico sem evidente origem intra-abdominal cirurgicamente tratável. A peritonite bacteriana espontânea é uma das complicações graves em pacientes com cirrose e ascite. Sem tratamento antibiótico precoce, esta complicação é associada com alta taxa de mortalidade. Portanto, o diagnóstico precoce e tratamento de peritonite bacteriana espontânea são necessários para a sobrevivência. O reagente de esterase de leucócitos pode rapidamente diagnosticar a peritonite bacteriana espontânea.ObjetivoEste estudo teve como objetivo descobrir a acurácia diagnóstica do teste com tiras de esterase de leucócitos para o diagnóstico de peritonite bacteriana espontânea.MétodosUm estudo transversal hospitalar unicêntrico foi realizado entre julho de 2013 e agosto de 2014 em crianças com cirrose hepática e ascite que foram admitidas no Departamento de Gastroenterologia Pediátrica no Hospital de Nemazee afiliado à Universidade de Ciencias Médicas de Shiraz (Irã). Todos os pacientes foram submetidos a paracentese abdominal, e o líquido ascítico foi processado para contagem de células, teste de tira de reagente de esterase de leucócitos (Combiscreen SL10) e cultura. Peritonite bacteriana espontânea foi definida como tendo uma contagem de polimorfonucleares (PMN ≥250/m3) no líquido ascítico. Sensibilidade, especificidade, valor preditivo positivo negativo do teste de esterase de leucócitos foram calculados de acordo com a fórmula.ResultadosForam analisados um total de 150 amostras de líquido ascítico de pacientes cirróticos; (53,2%) eram do sexo masculino e sua média de idade (4,33±1,88 anos). A atresia biliar (n=44, 29,4%) e hepatite neonatal idiopática (n=29, 19,3%) foram as etiologias mais frequentes de cirrose. Além disso, dor abdominal (68,6%) e distensão (64%) foram as queixas mais comuns de apresentação. De todos os casos, 41 (27,35%) foram diagnosticados com peritonite bacteriana espontânea (PMN ≥250/mm3). A sensibilidade e especificidade do teste de reagente de esterase de leucócitos segundo PMN ≥250mm3 foi de 87,80% e 91,74% e, para os resultados de cultura de líquido ascítico, de 88,23% e 77,44%. Valor preditivo positivo e valor preditivo negativo do teste em PMN ≥250mm3 foi de 80% e 95,23% e em casos com cultura positiva 33,33% e 98,09%, respectivamente. A eficiência do teste de reagente esterase de leucócitos no diagnóstico de peritonite bacteriana espontânea, de acordo com resultados de ≥250mm3 e cultura PMN, foi de 90,66% e 78,66%.ConclusãoO teste de tiras de esterase de leucócitos pode ser usado como um teste rápido para diagnóstico de peritonite bacteriana espontânea, devido a sua alta validade diagnóstica.

Effects of piperonyl butoxide on the toxicity of the organophosphate temephos and the role of esterases in the insecticide resistance of Aedes aegypti

Introduction The effects of piperonyl butoxide (PBO) on the toxicity of the organophosphate temephos (TE) and the role of esterases in the resistance of Aedes aegypti to this insecticide were evaluated. Methods A. aegypti L4 larvae susceptible and resistant to TE were pre-treated with PBO solutions in acetone at concentrations of 0.125, 0.25, 0.5, 1, and 2% for 24h and subsequently exposed to a diagnostic concentration of 0.02mg/L aqueous TE solution. The esterase activity of the larvae extracts pre-treated with varying PBO concentrations and exposed to TE for three time periods was determined. Results At concentrations of 0.25, 0.5, 1, and 2%, PBO showed a significant synergistic effect with TE toxicity. High levels of esterase activity were associated with the survival of A. aegypti L4 larvae exposed to TE only. Conclusions The results of the biochemical assays suggest that PBO has a significant inhibitory effect on the total esterase activity in A. aegypti larvae. .

Low malathion concentrations influence metabolism in Chironomus sancticaroli (Diptera, Chironomidae) in acute and chronic toxicity tests

Low malathion concentrations influence metabolism in Chironomus sancticaroli (Diptera, Chironomidae) in acute and chronic toxicity tests. Organophosphate compounds are used in agro-systems, and in programs to control pathogen vectors. Because they are continuously applied, organophosphates often reach water sources and may have an impact on aquatic life. The effects of acute and chronic exposure to the organophosphate insecticide malathion on the midge Chironomus sancticaroli are evaluated. To that end, three biochemical biomarkers, acetylcholinesterase (AChE), alpha (EST-α) and beta (EST-β) esterase were used. Acute bioassays with five concentrations of malathion, and chronic bioassays with two concentrations of malathion were carried out. In the acute exposure test, AChE, EST-α and EST-β activities declined by 66, 40 and 37%, respectively, at 0.251 µg L-1 and more than 80% at 1.37, 1.96 and 2.51 µg L-1. In chronic exposure tests, AChE and EST-α activities declined by 28 and 15% at 0.251 µg L-1. Results of the present study show that low concentrations of malathion can influence larval metabolism, indicating high toxicity for Chironomus sancticaroli and environmental risk associated with the use of organophosphates.

LEAF-CUTTING ANTS Acromyrmex niger SMITH, 1858(HYMENOPTERA; FORMICIDAE) USED AS BIOINDICATORS OF AGROTOXICS RESIDUES / Utilización de hormigas cortadoras Acromyrmex niger Smith, 1858 (Hymenoptera; Formicidae) como bioindicadores de residuos de agrotóxicos

Despite the condition of leaf-cutting ant pests in agroecosystems, it is undeniable the benefits they can bring in certain situations or environments. The leaf-cutting ants of the genus Acromyrmex attack mainly leaves of vegetables and fruit trees exposing not only to the agrochemicals used for their control as well as to those used for the control of other pests. Due to the bioindicator potential of environmental quality of the ants and their frequent exposure to agrochemicals such as organophosphates, neonicotinoids and growth regulators insecticide used for pest control, it is necessary to study the sublethal effects that these pesticides may cause. The electrophoresis technique was used to study the activity of esterase isozymes involved in the metabolism of xenobiotics of A. niger , combined with changes in the expression of isozymes after contamination by pesticides. A. niger showed eight regions of esterase activity, which were called EST-1, EST-2, EST-3, EST-4, EST-5, EST-6, EST-7 and EST-8 according to the electrophoretic mobility. As the specificity to ± and b-naphthyl acetate substrates, the Est- 7 and Est-8 may be ified as ±-esterase and the others as ±b esterases. EST-5 is considered an enzyme of the type cholinesterase II and the others are of the type carboxilesterase. The electrophoretic analysis showed partial inhibition to all esterases subjected to the contact with Malathion organophosforate at the concentrations 1 x 10-3 % and 5 x 10-3 %, which may be considered as a biomarker for the presence of residues of this insecticide in the environment. The regression analysis for sublethal effects of the tested pesticides demonstrated correlation between dose and mortality only for Thiametoxam neonicotinoid pesticide.

A pesar de la condición de plaga de las hormigas cortadoras en agroecosistemas, no se pueden negar los beneficios que ellas pueden traer en determinadas situaciones o en determinados ambientes. Las hormigas cortadoras del género Acromyrmex atacan principalmente las hojas de hortalizas y hojas de plantas fructíferas, exponiéndose no solo a los agroquímicos utilizados para su control sino también a aquellos utilizados en el control de otras plagas. Debido al potencial bioindica-dor de la calidad ambiental de las hormigas y su frecuente exposición a los agrotóxicos como organofosforados, neonicotinoides y reguladores de crecimiento, utilizados en el control de plagas, es necesario el estudio de los efectos subletales que estos agrotóxicos pueden causar. La técnica de electroforesis fue utilizada para evaluar la actividad de las isoenzimas esterasas involucradas en el metabolismo de xenobióticos de A. niger , asociada a las alteraciones en la expresión de las isoenzimas después de la contaminación con pesticidas. Acromyrmex Niger demostró ocho regiones de actividad esterasa, las cuales fueron denominadas EST-1, EST-2, EST-3, EST-4, EST-5, EST-6, EST-7 e EST-8 de acuerdo con la movilidad electroforética. En cuanto a la especificación de los substratos ± y b-nafitl acetato, las EST-7 e EST-8 son clasificadas como ±- esterasa y las demás ±b esterasas. EST-5 es considerada una enzima del tipo colinesterasa II y las demás son carboxilesterasas. El análisis electroforético presentó inhibición parcial para todas las esterasas sometidas al contacto con malathion en las concentraciones 1 x 10-3 % e 5 x 10-3 %, que puede ser considerado un biomarcador para la presencia de residuos de este insecticida en el ambiente. El análisis de regresión para el efecto subletal de los pesticidas evaluados demostró correlación entre la dosis y la causa de muerte solo para el pesticida neonicotinoide thiametoxam.

Analysing deltamethrin susceptibility and pyrethroid esterase activity variations in sylvatic and domestic Triatoma infestans at the embryonic stage

The aim of the present work was to study the deltamethrin susceptibility of eggs from Triatoma infestans populations and the contribution of pyrethroid esterases to deltamethrin degradation. Insects were collected from sylvatic areas, including Veinte de Octubre and Kirus-Mayu (Bolivia) and from domiciliary areas, including El Palmar (Bolivia) and La Pista (Argentina). Deltamethrin susceptibility was determined by dose-response bioassays. Serial dilutions of deltamethrin (0.0005-1 mg/mL) were topically applied to 12-day-old eggs. Samples from El Palmar had the highest lethal dose ratio (LDR) value (44.90) compared to the susceptible reference strain (NFS), whereas the Veinte de Octubre samples had the lowest value (0.50). Pyrethroid esterases were evaluated using 7-coumaryl permethrate (7-CP) on individually homogenised eggs from each population and from NFS. The El Palmar and La Pista samples contained 40.11 and 36.64 pmol/min/mg protein, respectively, and these values were statistically similar to NFS (34.92 pmol/min/mg protein) and different from Kirus-Mayu and Veinte de Octubre (27.49 and 22.69 pmol/min/mg protein, respectively). The toxicological data indicate that the domestic populations were resistant to deltamethrin, but no statistical contribution of 7-CP esterases was observed. The sylvatic populations had similar LDR values to NFS, but lower 7-CP esterase activities. Moreover, this is the first study of the pyrethroid esterases on T. infestans eggs employing a specific substrate (7-CP).

Malation vs. Aedes aegypti (Linnaeus) (Diptera: Culicidae) de diferentes regiones de Venezuela / Malathion vs. Aedes aegypti (Linnaeus, Diptera: Culicidae) from different regions of Venezuela

En Venezuela, el malation ha sido ampliamente usado en forma continua en programas de control de aedes aegypti. Por tal motivo se realizó un estudio en mosquitos provenientes de zonas urbanas con alta casuística de dengue de los estados: Amazonas, Aragua, Bolívar, Lara, Mérida y Zulia, para determinar el status de susceptibilidad en este vector al malatión, en comparación con la cepa susceptible referencial, Rockefeller (Rock). Se hicieron bioensayos en botellas tratadas con el insecticida malatión evaluando la dosis diagnóstica 100ug/mL en 30 minutos y ensayos bioquímicos en microplacas para determinar mecanismos metabólicos asociados al status frente al insecticida. Los resultados de los bioensayos mostraron que existe susceptibilidad a malatión, lo cual fue confirmado por pruebas bioquímicas. Sin embargo, se encontraron diferencias significativas entre todas las cepas evaluadas con valores de P<0,005 para esterasas alfa (α), esterasas beta (β) y acetilcolinesterasa normal (Ache) y acetilcolinesterasa inhibida (Achei). La prueba de comparación de medias de Bonferroni encontró similitud entre la cepa Rock, mazonas y Lara para esterasas α y β. Se encontró similitud de la cepa Rock con las cepas de Bolívar y Zulia para las pruebas con Ache y Achei. Este estudio concluye que el malatión mostró su potencial de uso en el control del vector del dengue de las localidades evaluadas.

In Venezuela, malathion has been widely used continuously in control programs for Aedes aegypti. Therefore, a study in mosquitoes from urban areas with high dengue casuistry in the states of Amazonas, Aragua, Bolivar, Lara, Merida and Zulia was conducted to determine the status of this vector susceptibility to malathion, compared with the reference susceptible strain, Rockefeller. Bioassays were done on bottles treated with the insecticide malathion, 100ug/mL evaluating the diagnostic doses in 30 minutes and biochemical assays in microplates were performed to determine metabolic mechanisms associated with status against insecticide. The bioassay results showed that there is malathion susceptibility, which was confirmed by biochemical tests. However, significant differences were found among all strains assessed values of P<0.005 for esterases alpha (α), beta esterases (β) and standard acetylcholinesterase (AChe) and inhibited acetylcholinesterase (Achei). The mean comparison test of Bonferroni showed similarity between the strains Rock, Amazon and Lara for esterases α and β. Similarity was found between the strains Rock, Bolivar and Zulia for the Ache and Achei tests. This study concludes that malathion showed its potential use in controlling the dengue vector in the locations evaluated.

Effects of perinatal exposure of lithium on neuro-behaviour of developing mice offspring.

Lithium (Li) was given to female Swiss-Webster strain mice at the doses of 15 and 30 mg/kg body weight in their drinking water. Treatment started from the first day of pregnancy until the postnatal day fifteen of delivery. Thereafter, the dams were switched to plain tap water. All offspring were subjected to various tests. The rate of body weight gain was relatively slower in Li exposed pups. Furthermore, the opening of eyes and appearance of body hairs in Li exposed pups were also slower as compared to the controls. The sensory motor reflexes in Li exposed pups were found to be affected in a dose-dependent manner. Significant relative changes were also noticed in the levels of acid and alkaline phosphatases in the liver, and acetylcholinesterase in the brain tissues of the Li exposed developing offspring in a dose-dependent manner. ‘Locomotor Activity Test’ was performed in the male offspring only which showed a significant suppressive effect on most of the elements of this test due to Li exposure. The present Li effects in the offspring are possibly via in utero action and/or via mother’s milk.

Isozymic variations in specific and nonspecific esterase and its thermostability in silkworm, Bombyx mori L.

Esterase isozymic variations were documented in the haemolymph of developed multivoltine and bivoltine silkworm breeds during unfavorable seed crop seasons of May – September using α- and β- napthylacetate separately to identify specific and nonspecific esterase having thermotolerant potentiality. Variations existed in the isozyme pattern with three bands (Est-2, 3 and 4) in pure Nistari race and other developed multivoltine and bivoltine breeds. Est-2 and Est-3 were non-specific esterases as they were observed when both α- and β-napthylacetate was used as substrates separately. Est- 4 band was observed only with α-napthylacetate as substrate and was therefore confirmed to be specific α-esterase band in the haemolymph of silkworm, Bombyx mori L. Zymograms showed that the non-specific esterase band (Est-3) with Rf of 0.43 and specific α-esterase band (Est-4) with Rf of 0.32 predominately withstood a temperature of 70 ± 2oC for a duration of 10 min and were confirmed as thermostable esterases in haemolymph of silkworm, Bombyx mori L. This also categorized the presence of thermostable esterases in developed multivoltine and bivoltine breeds of silkworm, even though the qualitative activity was more in the former than the latter. The qualitative presence of thermostable esterases and their activity could be adopted as an indicative biochemical marker in relation to thermotolerance in silkworm.

Caracterización parcial de la actividad de esterasas en una cepa de Aedes aegypti resistente a temefos / Partial characterization of esterase activity in a temephos-resistant Aedes aegypti strain

Introducción: las enzimas esterasas han sido identificadas como mecanismo de resistencia a temefos en Aedes aegypti de Cuba, larvicida más utilizado en el mundo. Objetivo: caracterizar parcialmente la actividad de esterasas en larvas expuestas y no expuestas a dosis subletales de temefos en una cepa de Aedes aegypti resistente a este insecticida. Métodos: se utilizó una cepa de Aedes aegypti de referencia susceptible (Rockefeller) y otra resistente a temefos (SANtemF11). Se expusieron las larvas de la cepa SANtemF11 a la concentración letal 90 (CL90) de temefos (1 ppm), 10 % de larvas sobrevivientes a las 24 h (SANtem [24 h]) se transfirieron a agua limpia y sin exposición a insecticidas por otras 24 h (SANtem [48 h]). Se caracterizó de modo parcial, en estas larvas, la actividad de esterasas a través de ensayos bioquímicos y electroforesis en gel de poliacrilamida. Se estimó por duodecil sulfato de sodio (SDS-PAGE) el peso molecular de la esterasa (Est. A4). Resultados: la actividad de esterasas en la cepa SANtemF11 resultó significativamente mayor que en Rockefeller. Se observó una disminución significativa de la actividad de esterasas en las larvas sobrevivientes (SANtemF11 [24 h]), la cual se recuperó 24 h después sin exposición a temefos. En el zimograma se observó que en 10 % de las larvas sobrevivientes a temefos, solo apareció incrementada la banda de esterasa A4, en comparación con las observadas en SANtemF11. El peso molecular estimado de la esterasa A4 fue de 58 kDa. Conclusiones: la presencia de una banda específica de esterasa (58 kDa), en las larvas sobrevivientes a la selección con temefos, confirma su papel en la resistencia a este insecticida. Diagnosticar la función de las esterasas en la resistencia a temefos, a través de ensayos bioquímicos, no debe realizarse en larvas expuestas a dosis subletales de este insecticida, para evitar falsos negativos.

Introduction: the esterase enzymes have been defined as the mechanism of resistance to temephos in Aeges aegypti in Cuba, which is the most used larvacide worldwide. Objective: to partially characterize the activity of esterases in exposed and non-exposed larvae at sublethal doses of temephos in an Aedes aegypti strain that is resistant to this product. Methods: a susceptible reference Aedes aegypti strain (Rockefeller) and another temephos-resistant strain (SANtemFII) were used. The larvae from SANtemF11 strain were exposed to lethal concentration 90 (LC90) of temephos (1 ppm); 10 % of the surviving larvae after 24 hours (SANtem[24 h] was moved to clean water, with no exposure to insecticide for 24 hours (SANtem [48 h]). The activity of esterases was partially characterized in these larvae through biochemical assays and gel-polyacrylamide electrophoresis. The molecular weight of esterase A 4 (ESt. A4) was estimated with the support of sodium duodecyl sulophate (SDS-PAGE). Results: the activity of esterases in SANtemF11 was significantly higher than in Rockefeller strain. Significant reduction of the activity of esterases in surviving larvae was observed (SANtemF11 [24 h], but it increased 24 h later without exposure to temephos. The zymogram showed that 10% of larvae that survived from temephos action, just the esterase A4 band increased if compared with those of SAntemF11. The estimated molecular weight of esterase A4 was 58 kDa. Conclusions: the presence of a specific band of esterase (58 kDa) in surviving larvae confirmed the role of these enzymes in insecticidal resistance. The diagnosis of the function of the esterases in resistance to temephos through biochemical tests should not be made in larvae exposed to sublethal doses of this insecticide, in order to avoid false negatives.

Influence of Rivastigmine transdermal on butyrylcholinesterase levels in patients with Alzheimers disease / A influência da rivastigmina transdérmica nos níveis de butirilcolinesterase em pacientes com doença de Alzheimer

Cholinesterase inhibitors (ChE-Is) are among the main drugs approved for the treatment of Alzheimer's disease (AD). Rivastigmine in the form of a transdermal patch is an alternative delivery method, and can give greater treatment compliance. Objectives: To conduct a preliminary assessment of the neurocognitive and biological effects of oral and transdermal Rivastigmine in patients with AD and to identify a potential biological marker and demonstrate a possible relationship between esterase levels and behavioral scores of AD patients. Methods: Forty patients with AD were treated with cholinesterase inhibitors (ChE-Is), evaluated using the MMSE and NPI, and simultaneously sampled to determine their serum levels of AChE and BuChE for 180 days. Results: The differences obtained between oral and transdermal forms, as assessed by the MMSE and NPI scores of the AD patients, were not significant at the three time points examined (0, 90, and 180 days). However, serum BuChE levels of the transdermal group differed significantly (p<0.0004) compared with those of the oral group at 90 days. Conclusion: Use of a transdermal ChE-I, rivastigmine tartrate significantly reduced BuChE levels in the AD patients studied.

Os inibidores das colinesterases estão entre as principais drogas aprovadas para tratamento da doença de Alzheimer (DA). Rivastigmina na forma de adesivo transdérmico é um método alternativo de liberação e pode fornecer uma maior aderância ao tratamento. Objetivos: Conduzir uma abordagem preliminar dos efeitos neurocognitivos e biológicos da rivastigmina oral e transdérmica em pacientes com DA e identificar um potencial marcador biológico e demonstrar uma possível relação entre níveis de esterases e escores de comportamento de pacientes com DA. Métodos: Quarenta pacientes com DA com inibidores de colinesterases foram avaliados usando o MEEM e o INP e colhidas amostras para determinar seus níveis séricos de AChE e BuChE por 180 dias. Resultados: As diferenças obtidas entre as formas oral e transdérmica, avaliadas pelo MEEM e INP não diferiram em três ocasiões (0, 90 e 180 dias). Todavia, os níveis de BuChE no grupo transdérmico diferiu significativamente (p<0.0004) comparados ao grupo de administração oral em 90 dias. Conclusão: O uso do tartarato de rivastigmina, forma transdérmica reduziu significativamente os níveis de BuChE nos pacientes estudados com DA.

Línea básica de susceptibilidad y efecto sinérgico a los insecticidas malation y pirmifos-metil en adultos de Culex spp. Say, 1823 (Díptera: Culicidae) del municipio Mario Briceño Iragorry del estado Aragua, Venezuela / Baseline susceptibility and synergistic effect to the insecticides malathion and pirmiphos-methyl in adults of Culex spp. Say, 1823 (Diptera: Culicidae) Mario Briceño Iragorry municipality, Aragua state, Venezuela

Se evaluó la respuesta a los insecticidas organofosforados malatión y pirimifos-metil en poblaciones de campo de adultos Culex spp. del Municipio Mario Briceño Iragorry del Estado Aragua. Se realizaron los bioensayos en botellas tratadas con insecticidas. La determinación del tiempo-mortalidad, permitió obtener la línea base de susceptibilidad de los insecticidas evaluados. Se definió la dosis diagnóstica como la menor dosis que mata el mayor porcentaje de los insectos expuestos. El número de sobrevivientes a los 60 minutos de exposición a dichas dosis fue considerado como criterio de resistencia definiéndose dicho tiempo, como el umbral de resistencia. Los mosquitos resultaron resistentes a malatión a las concentraciones 0,1; 1 y 5 μg/mL con valores de TL100= 90; 90 y 75 minutos respectivamente y susceptibles a pirimifos-metil a las concentraciones 1 y 5 μg/mL con valores TL100= 30 y 15 minutos respectivamente, a la concentración 0,1 μg/mL se obtuvo TL100= 75 lo cual no se estimó por ser una dosis baja. Las concentraciones 5 μg/mL de malatión, 0, μg/ml de pirimifos-metil se sugieren como dosis referenciales. Se identificaron mecanismos de resistencia con el sinergista butóxido de piperonilo (PB) a las concentraciones de malatión 1 y 5 μg/mL con FS = 2 y 2,5 respectivamente. Las enzimas de multifunción oxidasa (MFO) juegan un papel importante en la resistencia al insecticida malation. Los resultados obtenidos, aportan información básica para futuros programas de control de Culex spp., si fuese necesario, debido a su importancia como plaga de ambientes turísticos y como vector de Filariasis (bancroftiasis) y Fiebre del Nilo.

We evaluated the response to the organophosphorus insecticides malathion and pirimiphos-methyl in field populations of adult Culex spp. Mario Briceño Iragorry Municipality Aragua state. Bioassays were carried out using the bottles treated with insecticides. The time-mortality determination allowed us to obtain the baseline susceptibility of the insecticides evaluated. Diagnostic dose was defined as the lowest dose that kills the highest percentage of exposed insects. Survivors after 60 minutes of exposure to these doses was considered as resistance criterion defining this time, as the threshold of resistance. The mosquitoes were resistant to malathion at concentrations 0.1; 1 and 5 μg/mL with values of TL100= 90; 90 and 75 minutes respectively and susceptible to pirimiphos-methyl at concentrations 1 and 5 μg/mL with values of TL100= 30 and 15 minutes respectively, was obtained at the concentration 0.1 μg /mL TL100= 75 which was not considered to be a low dose. The concentrations 5 μg /mL of malathion, 0.1 μg / ml of pirimiphos-methyl are suggested as a reference dose. Resistance mechanisms were identified with the synergist piperonyl butoxide (PB) at concentrations of malathion 1 and 5 μg/mL with FS = 2 and 2.5 respectively. Multifunction oxidase enzymes (MFO) play an important role in resistance to the insecticide malathion. The obtained results provide basic information for future monitoring programs of Culex spp, if necessary, due to its importance as a pest of tourist environments and as a vector of Filariasis (bacroftiosis) and West Nile Fever.

Resistencia a insecticidas organofosforados en Aedes aegypti (Diptera: Culicidae) de Santiago de Cuba, 1997-2009 / Resistance to organophosphorus insecticides found in Aedes aegypti (Diptera: Culicidae) from Santiago de Cuba, 1997-2009

INTRODUCCIÓN: la resistencia a insecticidas organofosforados en Santiago de Cuba fue diagnosticada en Aedes aegypti (Linnaeus, 1762) en 1997, y alguno de ellos se han continuado utilizando hasta la fecha, de ahí la necesidad de conocer cómo ha variado la resistencia desde entonces, hasta fechas más recientes, año 2009. OBJETIVO: evaluar la resistencia a insecticidas organofosforados en larvas de Santiago de Cuba, colectadas en 2009 y su variación con respecto a 1997. Determinar la frecuencia en que aparece el mecanismo de resistencia, basado en la alta actividad de esterasas y su clasificación. MÉTODOS: se evaluó la resistencia a los insecticidas organofosforados, malation, pirimifos metil, fenitrotion, fention, temefos y clorpirifos en larvas mediante la metodología recomendada por la Organización Mundial de la Salud. El mecanismo de esterasas se determinó a través de ensayos bioquímicos y electroforesis en gel de poliacrilamida. RESULTADOS: las larvas de la cepa de Santiago de Cuba resultaron susceptibles a malation, pirimifos metil y fenitrotion y no hubo variación con los resultados obtenidos en una cepa de Santiago de Cuba de 1997, se observó moderada resistencia a fention y alta a temefos y clorpirifos. Al comparar estos resultados con los obtenidos en 1997, se observó un incremento de la resistencia a los tres insecticidas en el período 1997-2009. En la cepa de Santiago de Cuba de 2009 se demostró que las esterasas se encontraron con una alta actividad a una frecuencia de 0,7. Se observó la presencia de una esterasa tipo B amplificada, con un valor de movilidad relativa de 0,95 cm, la cual no se encontró en la cepa susceptible de referencia. CONCLUSIONES: la resistencia a insecticidas y sus mecanismos es un fenómeno sumamente variable, aun en la misma especie sometida a distintas intensidades de aplicación de insecticidas, de ahí que su monitoreo constante de forma local y en el tiempo sea una necesidad para un programa de control de vectores.

INTRODUCTION: resistance to organophosphorus insecticides was diagnosed in Aedes aegypti (Linnaeus, 1762) from Santiago de Cuba in 1997 and some of them are still used up to date; hence the need of ascertaining how the insecticidal resistance has changed in recent times, particularly in 2009. OBJECTIVE: to evaluate the resistance to organophosporus insecticides in larvae from Santiago de Cuba collected in 2009, and its variation in comparison with that observed in 1997; and to determine the frequency of occurrence of resistance mechanisms on the basis of high esterase activity and its classification. METHODS: resistance to organophosphorus insecticides such as malathion, pirimiphos, methyl, phenitrotion, phention, temephos and clorpiriphos in larvae by using the WHO recommended methodology. The esterase mechanism was identified through biochemical assays and polyacrylamide gel electrophoresis. RESULTS: larvae from the Santiago de Cuba strain were susceptible to malathion, pirimiphos, methyl and phenitrothion; there was no variation with the results achieved in a Santiago de Cuba strain in 1997, moderate resistance to phenthion and high resistance to temephos and chlorpiriphos were observed. When comparing these results with those of 1997, it was noted that resistance to the three insecticides increased in the 1997-2009 period. In the Santiago de Cuba strain 2009, it was shown that esterase activity was very high at a rate of 0,7. The presence of an amplified type B esterase with relative mobility of 0.95 cm was detected, which did not exist in the reference strain. CONCLUSIONS: resistance to insecticides and its mechanisms are highly variable, even in the same species subjected to various intensities in the insecticidal use, therefore, it is necessary to constantly monitor both aspects at local level in the course of time, with a view to an effective vector control program.