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.

Cometabolic biodegradation of quizalofop-p-ethyl by Methylobacterium populi YC-XJ1 and identification of QPEH1 esterase

BACKGROUND: Quizalofop-p-ethyl (QPE), a unitary R configuration aromatic oxyphenoxypropionic acid ester (AOPP) herbicide, was widely used and had led to detrimental environmental effects. For finding the QPEdegrading bacteria and promoting the biodegradation of QPE, a series of studies were carried out. RESULTS: A QPE-degrading bacterial strain YC-XJ1 was isolated from desert soil and identified as Methylobacterium populi, which could degrade QPE with methanol by cometabolism. Ninety-seven percent of QPE (50 mg/L) could be degraded within 72 h under optimum biodegradation condition of 35°C and pH 8.0. The maximum degradation rate of QPE was 1.4 mg/L/h, and the strain YC-XJ1 exhibited some certain salinity tolerance. Two novel metabolites, 2-hydroxy-6-chloroquinoxaline and quinoxaline, were found by high-performance liquid chromatography/mass spectroscopy analysis. The metabolic pathway of QPE was predicted. The catalytic efficiency of strain YC-XJ1 toward different AOPPs herbicides in descending order was as follows: haloxyfop-pmethyl ≈ diclofop-methyl ≈ fluazifop-p-butyl N clodinafop-propargyl N cyhalofop-butyl N quizalofop-p-ethyl N fenoxaprop-p-ethyl N propaquizafop N quizalofop-p-tefuryl. The genome of strain YC-XJ1 was sequenced using a combination of PacBio RS II and Illumina platforms. According to the annotation result, one α/ß hydrolase gene was selected and named qpeh1, for which QPE-degrading function has obtained validation. Based on the phylogenetic analysis and multiple sequence alignment with other QPE-degrading esterases reported previously, the QPEH1 was clustered with esterase family V. CONCLUSION: M. populi YC-XJ1 could degrade QPE with a novel pathway, and the qpeh1 gene was identified as one of QPE-degrading esterase gene.

Partial purification and biochemical characterization of an alkaline esterase from Sorghumbicolor

Esterases are enzymes that present good potential for industrial applications since they catalyze the formation or cleavage of ester bonds in water-soluble substrates, and sorghumseeds can represent an alternative source of this enzyme. The extraction of esterase from sorghumseeds is an economical alternative to obtain an enzyme of great interest. Esterases may improve the quality or accelerate the maturation of cheeses, cured bacon and fermented sausages and may also resolve racemic mixtures. Recently, seed esterases have been the focus of much attention as biocatalysts. In some cases, these enzymes present advantages over animal and microbial lipases due to some quite interesting features such as specificity and low cost, being a great alternative for their commercial exploitation as industrial enzymes The esterase studied here was extracted from sorghumseeds and some of its biochemical properties determined using synthetic substrates (p-nitrophenyl butyrate, caprylate, laurate and palmitate). The enzyme presented optimum activity at pH 8.0 and was stable in all the pH ranges studied. The optimum temperature for its activity was 40ºC but it showed low stability at this temperature (40% relative activity). The values derived for Km and Vmax were 0.67mM and 125 U.mg-1, respectively, obtained using p-nitrophenyl butyrate as the substrate. The enzyme showed an increase in activity when K2HPO4was added to the reaction medium, but the ions Mn2+, CO+, Hg+and Fe2+strongly inhibited the enzyme activity. This enzyme showed a preference for the hydrolysis of short chain fatty acids. The characteristics of sorghumesterase are very similar to those of the microbial esterases used in detergent processing.

Perfil de suscetibilidade da população de Aedes aegypti (Diptera: Culicidae) da ilha de Santiago, Cabo Verde, a inseticidas / Susceptibility profile of Aedes aegypti from Santiago Island, Cape Verde, to insecticides

Em Cabo Verde, arquipélago situado na Costa Ocidental Africana, os primeiros casos de dengue ocorreram em 2009, com a notificação de mais de 21.000 casos, a maioria desses registrados na Ilha de Santiago. O mosquito Aedes aegypti foi identificado como vetor, e ações para seu controle, usando os inseticidas temephos (larvicida) e a deltametrina (adulticida), têm sido implementadas. Objetiva-se com esse trabalho avaliar o atual status de suscetibilidade a inseticidas e caracterizar os mecanismos de resistência nessa população. Amostras de A. aegypti da ilha de Santiago foram coletadas através de armadilhas de oviposição, para o estabelecimento de uma população a ser analisada. Foram realizados bioensaios do tipo dose diagnóstica, usando garrafas impregnadas com doses únicas dos adulticidas malathion (organofosforado), deltametrina (piretróide) e cipermetrina (piretróide), e bioensaios do tipo dose resposta, usando múltiplas concentrações dos inseticidas temephos (organofosforado), Bacillus thuringiensis sorovariedade israelensis (bactéria entomopatogênica) e diflubenzuron (inibidor de síntese de quitina). Para a investigação dos mecanismos de resistências, foram realizados testes bioquímicos com substratos específicos para quantificar a atividade das enzimas glutationa S-transferases, esterases (alfa, beta e PNPA) e oxidases de função mista, ligadas a detoxificação de xenobióticos, e a taxa de inibição da acetilcolinesterase ligada a insensibilidade do sítio alvo...

Cape Verde, an archipelago located on the West African Coast, recorded the first cases of dengue in 2009 in an epidemic with more than 21,000 reportedcases. The worst affected area was Santiago Island...

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).

Endophytic fungi producing of esterases: evaluation in vitro of the enzymatic activity using pH indicator

A sensitive and efficient colorimetric method was optimized for detection of esterase enzymes produced by endophytic fungi for development of High-Throughput Screening (HTS). The fungi were isolated and obtained previously from plant species of Cerrado and Atlantic Forest located in areas of environmental preservation in the State of Sao Paulo / Brazil, as part of the project "Chemical and biological prospecting endophytic fungi associated to plant species of Cerrado and Atlantic Forest". The compounds ethyl butyrate, ethyl acetate and methyl propionate were used as standards esters which were hydrolyzed by extracellular enzyme from endophytic fungi (EC. 3.1.1.1 -carboxylesterases) for production of carboxylic acids. Thus, the reduction of the pH increases the protonated indicator concentration (bromothymol blue), changing the color of the reaction medium (from blue to yellow), that can be observed and measured by spectrophotometry at 616 nm. The methodology with acid-base indicator was performed on 13 microorganisms, aiming Periconia atropurpurea asapotential source of esterase for biotransformation of short chain esters. The results also evidenced that this methodology showed to be efficient, fast, cheap, having low consumption of reagents and easy development, and can be applied to screen carboxylic-ester hydrolases in a large number of microorganisms.

Estado de la susceptibilidad de poblaciones naturales / Susceptibility of natural populations of dengue vector to insecticides in Colombia

Introducción. La resistencia fisiológica de las poblaciones naturales de Aedes aegypti a los insecticidas de uso en salud pública, disminuye la eficacia del control químico utilizado para interrumpir la transmisión de dengue durante los brotes y epidemias. Objetivo. Determinar el estado de la susceptibilidad al carbamato propoxur, a los organofosforados adulticidas malatión y fenitrotión y al larvicida temefos, de 13 poblaciones naturales de Ae. aegypti en ocho municipios: una de Bucaramanga y dos de Sabana de Torres en el departamento de Santander; dos de Girardot y dos de La Mesa en Cundinamarca; dos de Villavicencio y dos de Puerto López en el Meta; una de San José del Guaviare, en Guaviare, y una de Florencia en Caquetá. Materiales y métodos. Las pruebas de susceptibilidad consistieron en bioensayos estandarizados de la Organización Mundial de la Salud (OMS), 1981, y botellas de los Centers for Disease Control and Prevention (CDC), 1998. Se aplicaron pruebas colorimétricas para determinar los niveles de enzimas como posibles responsables de la reducción de la susceptibilidad a insecticidas organofosforados y al carbamato propoxur. Resultados. Todas las poblaciones presentaron susceptibilidad a malatión y propoxur. Para el temefos, solo cuatro de las trece localidades evaluadas registraron susceptibilidad. Con respecto a los posibles mecanismos de desintoxicación enzimática, en siete de las once poblaciones naturales se registraron valores superiores al punto de corte para esterasas inespecíficas, solamente en Bucaramanga se presentó un incremento en las enzimas monooxigenasas del citocromo P450, pero en ninguna población se registró acetilcolinestarasa modificada. Conclusiones. La susceptibilidad generalizada a los organofosforados adulticidas evaluados, indica que el malatión, el insecticida más utilizado en Colombia para interrumpir la transmisión del dengue, sigue siendo eficaz. Las particularidades de los resultados de la resistencia fisiológica a insecticidas de las comunas de un mismo municipio, evidenciaron un fenómeno localizado de manera específica en espacio y tiempo.

Introduction. Physiological resistance of natural population of Aedes aegypti to insecticides contribute to the decreased efficacy of chemical control as a main control strategy during dengue outbreaks. Objective. The susceptibility status of Ae. aegypti was assessed for the carbamate propoxur, the adulticide malathion and the larvicide temephos on 13 natural populations of Ae. aegypti immature forms were taken from 8 Colombian localities. These included the following: Bucaramanga (1), Sabana de Torres (2), Girardot (2), La Mesa (2), Villavicencio (2), Puerto López (2), San José del Guaviare (1) and Florencia (1). Materials and methods. Susceptibility tests mainly consisted of the standardized bioassay outlined by WHO (1981) and CDC bottles (1998). Colorimetric tests were undertaken to determine enzyme levels possibly responsible for the reduction of susceptibility to organophosphate and carbamate insecticides. Results. All specimens demonstrated susceptibility to malathion and propoxur insecticides. Four of the 13 populations revealed susceptibility to the temephos larvicide. Seven of 11 populations showed a limited increase in values for nonspecific esterase enzymes. The Bucaramanga population was the only one which showed an increase in the cytochrome P450 monooxygenases enzymes. Neither population was found with modified acetilcolinesterase. Conclusions. The widespread susceptibility to organophosphates used as adulticides indicated that malathion, the most used insecticide in Colombia, remains effective in interrupting the transmission of dengue. Physiological resistance to insecticides occurring in communities of a single township proved to be a localized phenomenon.

Impact of environmental temperatures on resistance to organophosphate insecticides in Aedes aegypti from Trinidad / Repercusión de las temperaturas ambientales sobre la resistencia de Aedes aegypti a los insecticidas organofosforados en Trinidad

OBJECTIVE: To examine the effects of increasing larval rearing temperatures on the resistance status of Trinidadian populations of Aedes aegypti to organophosphate (OP) insecticides. METHODS: In 2007-2008, bioassays and biochemical assays were conducted on A. aegypti larvae collected in 2006 from eight geographically distinct areas in Trinidad (Trinidad and Tobago). Larval populations were reared at four temperatures (28 ± 2ºC, 32ºC, 34ºC, and 36ºC) prior to bioassays with OP insecticides (fenthion, malathion, and temephos) and biochemical assays for esterase enzymes. RESULTS: Most larval populations reared at 28 ± 2ºC were susceptible to fenthion (>98% mortality) but resistant to malathion and temephos (< 80% mortality). A positive association was found between resistance to OP insecticides and increased activities of α- and β-esterases in larval populations reared at 28 ± 2ºC. Although larval populations reared at higher temperatures showed variations in resistance to OPs, there was a general increase in susceptibility. However, increases or decreases in activity levels of enzymes did not always correspond with an increase or decrease in the proportion of resistant individuals reared at higher temperatures. CONCLUSIONS: Although global warming may cause an increase in dengue transmission, based on the current results, the use of insecticides for dengue prevention and control may yet be effective if temperatures increase as projected.

OBJETIVO: Examinar los efectos del aumento de las temperaturas de desarrollo larvario sobre el estado de resistencia a los insecticidas organofosforados de las poblaciones de Aedes aegypti en Trinidad. MÉTODOS: En 2007 y 2008 se llevaron a cabo ensayos biológicos y bioquímicos en larvas de A. aegypti recogidas en el 2006 de ocho áreas geográficamente separadas en Trinidad (Trinidad y Tabago). Las poblaciones larvarias se desarrollaron en cuatro temperaturas (28 ± 2 ºC, 32 ºC, 34 ºC y 36 ºC) antes de los ensayos biológicos con insecticidas organofosforados (fentión, malatión y temefós) y los análisis bioquímicos para las enzimas de esterasa. RESULTADOS: La mayoría de las poblaciones larvarias que se desarrollaron a 28 ± 2 ºC fueron susceptibles al fentión (mortalidad > 98%) pero resistentes al malatión y al temefós (mortalidad < 80%). Se encontró una asociación positiva entre la resistencia a los insecticidas organofosforados y la mayor actividad de αy β-esterasas en las poblaciones larvarias que se desarrollaron a 28 ± 2 ºC. Aunque las poblaciones larvarias que se desarrollaron a temperaturas mayores mostraron variaciones en la resistencia a los organofosforados, hubo un aumento general de la sensibilidad. Sin embargo, los aumentos o las disminuciones en los niveles de actividad de las enzimas no siempre se correspondieron con un aumento o disminución en la proporción de individuos resistentes desarrollados a las temperaturas más altas. CONCLUSIONES: Aunque el recalentamiento del planeta puede causar un aumento de la transmisión del dengue, según los resultados de este estudio el uso de insecticidas para la prevención y el control del dengue todavía puede ser eficaz si las temperaturas aumentan según lo proyectado.

Determinación in vivo del papel de las enzimas esterasas y glutation transferasa en la resistencia a piretroides en Aedes aegypti (Diptera: Culicidae) / Determination in vivo of the role of esterase and gluthation transferase enzymes in pyrethroid resistance of Aedes aegypti (Diptera: Cullicidae)

Se realizó un estudio in vivo a través del uso de 2 sinergistas, el trifenil fosfato (TFF) inhibidor específico de esterasas y el ácido etacrínico (AE), inhibidor específico de la enzima glutation transferasa (GST), para determinar si estas enzimas eran responsables de la resistencia a piretroides en Aedes aegypti. Para el trabajo se utilizaron 2 cepas de Aedes aegypti resistentes a insecticidas, una cepa que fue seleccionada con temefos por 6 generaciones de selección (SAN-F6) y otra con deltametrina por 12 generaciones de selección con este insecticida (SAN-F12), ambas resultaron ser resistentes a insecticidas piretroides. Se demostró a través del uso de los sinergistas TFF y AE que las enzimas esterasas y GST son responsables de la resistencia a los piretroides en estas cepas. Esos resultados demuestran la existencia de un fenómeno de resistencia cruzada y multirresistencia, lo cual debe tenerse en cuenta en las estrategias de uso de insecticidas para el control de este vector.

An in vivo study of two synergists, that is, Triphenil phosphate -specific esterase inhibitor- and ethacrynic acid specific gluthation transferase inhibitor- was performed to determine if these enzymes were responsible for pyrethroid resistance of Aedes aegypti. To this end, two insecticide resistant Aedes aegypti strains were used, one strain selected with temephos by six selection generations (SAN-F6) and the other strain with delmamethrin by 12 selection generations (SAN-F12), being both strains resistant to pyrethroid insecticices. Through the use of TPP and EA synergists, it was proved that esterase and gluthation-s-transferase (GST) enzymes were responsible for pryrethroid resistance of these strains. These results showed the existence of cross-resistance and multidrug resistance, which should be taken into account for insecticide use strategies aimed at vector control.

Susceptibilidad y mecanismos de resistencia a insecticidas en Anopheles albimanus del sur de la Península de Yucatán, México / Susceptibility and inseticide resistance mechanisms in Anopheles albimanus from the southern Yucatan Peninsula, Mexico

OBJETIVOS: Diagnosticar la resistencia a insecticidas y sus mecanismos en Anopheles albimanus del sur de la Península de Yucatán (PY), México. MATERIAL Y MÉTODOS: La F1 de An. albimanus colectados durante noviembre-diciembre de 2005 en seis localidades de los municipios Othón P. Blanco en Quintana Roo y Calakmul en Campeche, fue sometida a pruebas de susceptibilidad con deltametrina, DDT, pirimifos metil y bendiocarb, y a pruebas bioquímicas para calcular los niveles de las enzimas involucradas en la resistencia. RESULTADOS: An. albimanus fue resistente al DDT y a deltametrina en las seis localidades con niveles elevados de glutatión S-transferasas (GSTs), monooxigenasas y esterasas, y a pirimifos metil en La Unión con una alta frecuencia de la acetilcolinesterasa (AChE) alterada. CONCLUSION: Las poblaciones de An. albimanus colectadas al sur de la PY son resistentes al DDT y deltametrina, y en La Unión además al pirimifos metil, con mecanismos basados en la AChE alterada para el pirimifos metil, GST para DDT, y monooxigenasas y esterasas para piretroides. Los resultados del presente estudio tienen importantes consecuencias prácticas para el control químico de An. albimanus en el sur de la PY.

OBJECTIVE: To diagnose susceptibility levels and insecticide resistance mechanisms in Anopheles albimanus from the southern Yucatan Peninsula (YP), Mexico. MATERIAL AND METHODS: F1 generation of An. albimanus females, collected from November to December 2005 in six villages in the Othon P. Blanco municipality in Quintana Roo and the Calakmul municipality in Campeche, were exposed to deltamethrin, DDT, pirimiphos-methyl and bendiocarb in susceptibility tests, as well as to biochemical assays in order to calculate the enzyme levels related to insecticide resistance. RESULTS: High levels of DDT and deltamethrin resistance were found in An. albimanus collected from the six villages, and a high resistance to pirimiphos-methyl was found in those from La Union, Quintana Roo. Biochemical assays showed high levels of glutathione S-transferase (GST), cytochrome P450 and esterases (with pNPA substrate) in all villages. The frequency of An. albimanus with altered acetylcholinesterase (AChE) was high in La Union (33 percent). CONCLUSIONS: The An. albimanus populations collected in the south of the YP are resistant to DDT and deltamethrin, whereas resistance to pirimiphos-methyl was significant only in those collected from La Union. The mechanisms explaining this resistance are based on high concentrations of GST, cytochrome P450 and esterasas, the former being responsible for DDT metabolism and the others for pyrethroid metabolism. The altered AChE was the mechanism correlated to pirimiphos-methyl resistance in La Union. The results of the present study have important practical consequences for the chemical control of An. albimanus in the south of the YP.

Characterization of esterases in tetragonisca angustula and tetragona clavipes (Hymenoptera Meliponinae)

Esterases form a large, diverse group of enzymes with wide, overlapping substrate specificities and patternsof inhibition. These enzymes occur in a large variety of isoforms encoded by distinct gene loci with ahigh genetic variability and temporal differences in expression that make them appropriate for studyingpopulation structure. In this work, we investigated the substrate specificity and pattern of esterase expressionin body parts of the stingless bees Tetragonisca angustula and Tetragona clavipes. Fourteen hives of T.angustula were collected in Cianorte and T. clavipes were collected from three colonies in Maringá, twocities in the southern Brazilian state of Paraná. The esterase electrophoretic patterns were determinedusing polyacrylamide gels. Seven bands of esterase activity were detected in T. clavipes (EST-1 to EST-7)and two bands in T. angustula (EST-1 and EST-2). There was variation in the tissue esterase activity of T.clavipes, with EST-6 occurring in the abdomen of workers and EST-7 occurring in cephalic/thoracic extracts.The differences in the number of esterase bands and substrate specificity were attributed to the number ofesterase loci involved in each species, and/or variation in substrates specificities. The variation seen hereshould be useful for determining the role of esterases in intermediate metabolism in the Trigonini, as wellas to use esterases as a genetic marker for this stingless bee.

Evidence for a natural hybrid of peacock bass (Cichla monoculus vs Cichla temensis) based on esterase electrophoretic patterns

Esterase (Est) and esterase-D (Est-D) electrophoretic patterns identified by starch gel electrophoresis of skeletal muscle protein extracts of 184 specimens of three species of peacock bass, locally known as tucunares (Cichla monoculus, C. temensis and Cichla sp), plus four specimens of a supposed hybrid (C. monoculus vs C. temensis), collected from the Central Amazon, were examined to determine if they could aid in identifying a supposed hybrid between C. monoculus and C. temensis. Six zones of electrophoretic activity were found with these enzyme systems. The Est enzyme showed one zone of activity, formed by bands 1, 2 and 3, plus three zones of activity, presumably controlled by Est-1, 2 and 3 loci. The Est-D enzyme had two zones of activity, presumably controlled by Est-D1 and Est-D2 loci. Cichla monoculus and C. temensis shared band 2 and alleles Est-1(1), Est-2(1), Est-3(2), and Est-D1(1), and therefore these were useless for identifying hybrids between the two species. However, a probable hybrid pattern of bands 1, 2, and 3, presumably generated by a combination of pattern 12 from C. monoculus with pattern 23 from C. temensis, resulting from a possible cross between these two species, was detected. Although the Est-D2 locus cannot be considered an ideal diagnostic marker for identifying the supposed hybrid (C. monoculus vs C. temensis), as it is polymorphic, it proved to be useful for determining the origin of the hybrid, i.e., which parental species were involved in the hybridization process

Estudio de la resistencia en una cepa de Culex quinquefasciatus, procedente de Medellín, Colombia

Para conocer el estado de la resistencia en una cepa de Culex quinquefasciatus, procedente de una localidad de la ciudad de Medellín, Colombia, se determinaron los niveles de susceptibilidad y/o resistencia a 5 insecticidas organofosforados (malatión, metil-pirimifos, clorpirifos, temefos y fentión), 4 piretroides (cipermetrina, deltamemetrina, permetrina y lambdacialotrina) y un carbamato (propoxur). Se observó resistencia a todos los insecticidas organofosforados, aunque con valores relativamente menores para metil-pirimifos y fentión. No se encontró resistencia a los piretroides lambdacialotrina y cipermetrina, ni al carbamato propoxur, insecticidas que pueden ser muy útiles para el control de mosquitos de Colombia. Se demostró mediante el uso del sinergista piperonil butóxico que las oxidasas de función múltiple desempeñaron una función importante en la resistencia a los insecticidas organofosforados y piretroides. El uso del S.S.S. tributil fosfotritiado reveló que la sobreproducción de esterasas inespecíficas constituyó un mecanismo de resistencia para los insecticidas organofosforados, excepto metil-pirimifos y para los piretroides excepto lambdacialotrina. Este resultado debe tenerse en cuenta en las estrategias que se vayan a usar para el control de Culex quinquefasciatus de Colombia. Estos 2 mecanismos de resistencia no son responsables de la resistencia al carbamato propoxur. El análisis electroforético reveló la presencia de las esterasas B1, A6 y B6, que presumimos tienen una función importante en la resistencia.

The levels of susceptibility and/or resistance to 5 organophosphate insecticides (malathion, methyl-pyrimifos, clorpirifos, temephos and fenthion), 4 pyrethroids (cypermethrin, deltamethrin, permethrin and lambda-cyhalothrin), and a carbamate (propoxur) were deternmined in order to know the state of resistance in a strain of Culex quinquefasciatus from a locality of the city of Medellín, Colombia. Resistance to all organophosphate insecticeides, though with relatively lower values for methyloirimifos and fenthion, was observed. No resistance to lambda-cyhalothrin and cypermethrin or to propoxur was found. These insecticides may be useful for the control of mosquitoes in Colombia. It was demonstrated by using the piperonil butoxide sinergist that the oxidases of multiple function played an important role in the resistance to organophosphate insecticides and pyrethroids. The utilization of S.S.S. tributyl phosphotritiate revealed that the superproduction of unspecific esterases was a mechanism of resistance to organophosphate insecticides, except methyl-pirimifos and for perythroids, exceptlambda-cyhalothrin. This result should be taken into consideration for the strategies to be used to control Culex quinquefasciatus in Colombia. These two mechanism of resistance are not responsible for the resistance to propoxur. The electrophoretic analysis showed the presence of esterases B1, A6 and B6, which seem to have an important function in resistance.

Contribución al estudio de pruebas diagnósticas: sobre sus índices fijos y valores predictivos / Contribution to study of diagnostic tests: about it fixed indexes and predictive values

Las pruebas diagnósticas son una herramienta común y útil en la práctica clínica, cuya eficiencia se mide a través de su sensibilidad, específicidad y valores predictivos, los cuales se calculan después de comparar los resultados de la prueba con un patrón de referencia. Aunque las guías básicas para medir validez de una prueba son bien conocidas, se usó como modelo una publicación sobre el comportamiento de tres pruebas diagnósticas en 990 niños con infección del tracto urinario, para examinar las implicaciones que tienen algunas de las exigencias de tales guías en relación con los llamados índices fijos. Después de estudiar los índices fijos del grupo total y de cada una de las tres categorías clínicas de la enfermedad, las variaciones de éstos plantean la interrogante sobre cuáles índices usar para el cálculo de los valores predictivos de las pruebas. Se muestra que la fuente de distorsiones o mal uso de los valores predictivos en la medición de eficiencia de pruebas diagnósticas dependen del espectro de la enfermedad, teniendo presente la historia natural de ésta, además de las estrategias usadas en la referencia de pacientes al momento del estudio de la prueba y de los objetivos definidos para su uso clínico posterior

Monitoring resistance gene frequencies in Malaysian Culex quinquefasciatus Say adults using rapid non-specific esterase enzyme microassays.

The ability to identify the occurrence of different resistance genotypes in field populations of mosquito is considered important for the purpose of optimising chemical control operations. The recent development of rapid microassays of enzymes responsible for resistance has provided a means for rapidly assessing the genetic background of target mosquito populations. This concept is the topic of investigation in this study. Non-specific esterase activity, which is responsible for the resistance to organophosphates in Malaysian Culex quinquefasciatus Say adults, was determined in 3 field populations from Kuala Lumpur City using rapid enzyme assay. The optical density results were used to estimate the genotypic frequencies of the populations. Subsequently, time-dependent changes in the various frequencies were determined. Such techniques allowed rapid assessment of resistance genotypes for decision-making and its possible use in insect control merits further investigation.

Estudo das junçöes mioneurais de cobaia através das técnicas de esterase inespecífica e azul de metileno / Study of myoneural junctions of guinea-pig through non-specific esterase and methylene blue techniques

As junçoes mioneurais e o calibre das fibras musculares de músculos da parede abdominal de cobaia foram medidas após serem corados com esterase inespecífica e Azul de Metileno. Demonstrou-se que existe correlaçao direta entre diâmetro da fibra muscular e dimensao da sua junçao mioneural. Nao existe, entretanto, correspondência precisa entre os valores obtidos nos dois métodos utilizados. Este último aspecto deixa claro que, embora importante para o estudo da morfologia das junçoes mioneurais, o método para demonstraçao da esterase inespecífica nao se presta para estudos morfométricos ou onde se objetiva estudar os axonios pré-terminais, já que ele provoca acentuada retraçao do tecido e nao marca os axonios.

Relación entre la actividad de esterasas y la resistencia a malatión en una cepa de Tribolium Castaneum / Relation activity from enterase and t.castaneum malathion resistant

Se estudiaron comparativamente las esterasas de las cepas de Tribolium castaneum ULP (suceptible) y ML (resistente a malatión). La actividad de esterasas que hidrolizan acetato de * y ß naftol presentes en homogenatos de T. castaneum fue mayor en la cepa susceptible ML, mientras que la actividad frente al acetato de tiofenilo fue similar en ambas cepas. La distribución subcelular de la actividad esterástica frente al PTA mostró diferencias en ambas cepas. En los homogenatos de insectos de la cepa susceptible se observó que a mayor velocidad de centrifugación el sobrenadante resultó considerablemente más activo en la hidrólisis de acetato de tiofenilo. Se supone que ese resultado se debe a la presencia de un inhibidor endógeno no dializable, de peso molecular tal que puede ser parcialmente precipitado a 10.000g y en mayor medida a 100.000g. Los zimogramas de esterasas no inhibibles por eserina separadas por electroforesis en gel de poliacrilamida y reveladas con * y ß naftilacetato, mostraron, en todos los casos, una única banda que fue clasificada como aril o acetil esterasa. Los resultados descriptos encuadran en la teoría de la aliesterasa mutada, postulada para casos de resistencia al malatión, descriptos en otas especies de insectos

ESD, ACP and GLO frequencies in southern Brazil, with a noton heterozygosity levels

Os polimorfismos genéticos da fosfatase ácida, esterase D e glioxalase I foram investigadas em uma amostra de 363 indivíduos adultos brancos habitantes do Estado do Rio Grande do Sul. As freqüências gênicas observadas foram: ACP*A: 0,27; ACP*C: 0,03; ESD*2: 0,13; GLO*1: 0,42. Um alelo raro (ESD*3) foi encontrado em um indivíduo e seu filho. Utilizando-se dados deste e de estudos prévios calculou-se, para a populaçäo de Porto Alegre, o grau de heterozigosidade médio para respectivamente 13 e 20 sistemas eletroforéticos. Os níveis observados säo da mesma ordem de magnitude dos encontrados na América do Norte, apesar do grau mais elevado de mistura racial observado entre brasileiros