Toxicological evaluation of different pesticides in Tetragonisca angustula Latreille (Hymenoptera, Apidae)

The stingless bee Tetragonisca angustula is an important pollinator of different agricultural and native crops. This study evaluated changes in the relative activity of esterases and critical electrolyte concentration in brain cells after exposure to pesticides malathion and thiamethoxam. Lethal concentration 50% showed greater toxicity of thiamethoxam in relation to malathion. Esterases EST-3 and EST-4 (carboxylesterase) were partially inhibited after contamination by contact and ingestion of malathion and contamination by contact with thiamethoxam, suggesting participation of these esterases in the metabolization of these compounds. The lowest critical electrolyte concentration (CEC) was found after contamination by malathion ingestion (0.15 M), indicating changes in gene expression. The alterations observed in the intensity of EST-3 and EST-4 and the chromatin structure indicate that pesticides can act in gene expression and be used as biomarkers of contaminant residues. Furthermore, knowing the susceptibility of T. angustulabees to pesticides, it would be possible to use this species for biomonitoring environmental quality in preserved areas and agroecosystems.(AU)

Mechanisms of acaricide resistance in ticks

Background: In several countries, including Brazil, the livestock industry plays a key role in the country's economy. Brazil has the second largest bovine herd in the world and the biggest commercial herd. Ticks are an ongoing problem for both large operation cattle producers and small family farmers. Rhipicephalus microplus causes expressive losses in cattle breeding, since it occurs in important beef production zones like South America, Africa, and Oceania. Some of the negative consequences of tick infestation to cattle breeding are anemia, loss in milk and beef production, and transmission of Babesia bovis and B. bigemina. Significant losses are caused by the cattle tick (R. microplus) in several regions of the world, costing around US$ 3.3 billion per year to the Brazilian livestock industry alone. The tick control methods are mainly based on synthetic acaricides. However, the improvement of current tick control requires the identification of new molecular targets in tick physiology and development of molecule compounds to target important physiology pathways. The strategies proposed to address this issue are expand the knowledge about the molecules involved in the detoxification of chemicals to enhance the efficacy of the acaricides as well as to develop new compounds for chemical control. Review: Tick control is currently based on chemical acaricides; however, effective control and prevention of tick infestation remain distant goals. In recent decades, a progressive decrease in the efficiency of acaricides due to drug resistance has been observed. Acaricide resistance is an evolutionary adaptation, which implies the existence of behavioral and physiological mechanisms that allow the survival of resistant individuals. Four resistance mechanisms are described: behavioral resistance, reduced drug penetration, target site insensitivity and increased drug detoxification. Augmented drug detoxification may be due to increased activity of enzymes or transporters due to increased gene expression or mutations in some genes. Research focus on mechanisms of acaricide resistance in ticks characterized detoxification pathways based on (1) increased activity of enzymes (cytochrome p450, esterase and GST) which play a role in biochemically altering acaricides towards decreased toxicity and, (2) enhanced excretion of the modified less toxic compounds. To bypass the current problems, a better understanding of the biology, physiology, and molecular biology of the mechanisms of resistance to acaricides is fundamental to prolong their efficiency in controlling ticks. Moreover, identifying the genes and proteins associated with resistance can support in the development of more sensitive diagnostic methods to identify acaricide resistance, as well as improving control strategies. Discussion: In the last years, many researchers have been studying resistance mechanisms and important advances have been made which showed that, in several tick species, ABC transporters, esterases, P-450 cytochromes and glutathione-S-transferases participate in acaricide resistance. The characterization of the alterations in the targets in tick physiology and identification of new drugs with potential to tick control are crucial goals to increase tick control

Evaluation of the insecticidal activity of Solanum mammosum (L) fruit extract against Drosophila melanogaster

Solanum mammosum (L.) is known as a poisonous plant and has been extensively used for anti-cancer, antifungal, antibacterial properties. However, the effectiveness of Solanum mammosum against insects has not been fully reported yet. To assess the ability of Solanum mammosum fruit extract against insects, we used fruit fly (Drosophila melanogaster) as a model. In this study, our results showed that phytochemicals and many secondary metabolites were present in Solanum mammosum fruit extracts such as alkaloids, saponins, tannins, terpenoids, coumarins, cardiac glycosides, steroidstriterpenoids, flavonoids, and phenolic compounds. The total polyphenol and flavonoid content were also determined to be 275±1.89 mg GAE/g extract and 676±5.14 mg QE/g extract, respectively. Besides, Solanum mammosum fruit extract caused lethal to 2nd instar, reduced a pupae formation rate of Drosophila larvae. Interestingly, Solanum mammosum extract is more effective in the suppression of pupae eclosion than Ascend as a commercial pesticide. These phenomena might be associated with reduced accumulation of energy, including carbohydrates, lipids, and protein. In addition, the activities of esterases and phosphatases were inhibited in Drosophila by the intake of Solanum mammosum. The study concluded that Solanum mammosum has great potential as an insecticidal agent.

Comparative toxicity of fipronil, malathion, and thiamethoxam on the stingless bee Tetragonisca fiebrigi (Schwarz, 1938)

Stingless bees are important pollinators for various plant crops. We investigated the susceptibility of Tetragonisca fiebrigito sublethal concentrations of insecticides fipronil, malathion, and thiamethoxam (administered through contact and ingestion) by determining the LC50values after 24hoursof exposure and analyzing changes in the activity of esterase isoenzymes and the chromatin in brain cells. The LC50values showed that all three insecticides were highly toxic through contact and ingestion. Electrophoretic analysis revealed that the relative EST-4 (carboxylesterase) activity in T. fiebrigi was partially inhibited by malathion and fipronil ingestion. Moreover, the EST-4 band intensity was increased following high-concentration thiamethoxam (contact) exposure, indicating the increased relative activity of this isoenzyme to detoxify the compound. In the cytochemical analysis of brain cells, the critical electrolyte concentration (CEC) points for the control stingless bees and malathion ingestion-exposed and thiamethoxam-exposed (contact and ingestion) stingless bees were in the range of 0.20-0.30 M MgCl2, whereas that for malathion contact-exposed bees was 0.15 M MgCl2, indicating chromatin relaxation and suggesting an increase in gene expression. In conclusion, T. fiebrigistingless bees are susceptible to the insecticides tested, and the parameters analyzed may be used as biomarkers to detect the presence of these compounds.(AU)

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.

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)

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)

Genetic characterization of silkworm (Bombyx mori L. ) strains (Lepidoptera: Bombicidae) with different geographical originon the basis of isozyme markers

The aim of the present study was to assess genetics variations within and among strains with different geographical origin, belonging to Bulgarian germplasm bank of mulberry silkworm (Bombyx moriL.)and to establish their relationships using isoenzyme markers. Polyacrilamide gel electrophoresis (PAGE) was used to study the isoenzymes of nonspecific esterases (EST), malate dehydrogenase (MDH) and acid phosphatase (ACP) from haemolymph, phosphoglucomutase (PGM) and hexokinase (HK) from silk glands and alkaline phosphatase (ALP) from midgut of mulberry silkworm (Bombyx moriL.). Variability was evident in all of these enzyme systems among twelve strains from Bulgarian germplasm resources of B. mori.Total of nine lociwere detected. All of them (100%) were polymorphic. “Null” alleles in four lociwere determined. Intra-and inter-strain polymorphism was obtained. The degree of polymorphism ranged from 0% to 77.80%. Low levels of observed heterozygosity in comparison with the expected one have been calculated in almost all of strainsas well as deviations from Hardy-Weinberg equilibrium in almost all analyzed loci, resultingof excess of homozygotes. The value of FSTwas 0.4903. The dendrogram constructed with the values of genetic distance manifests that Romanian strain Cislau Tokay is formed one main clade while the rest strains studied (from Bulgaria, Japan, China, Vietnam, Spain, Syria and Egypt) are formed the other clade withsubclades. The genetic data of the tested strains from different geographical regions, would be used for identifying suitable parents for breeding programs with a view to yield improvement in this species with economic significance.

Diagnostic methods for identification of root-knot nematodes species from Brazil / Métodos diagnósticos usados na identificação de espécies do nematoide das galhas do Brasil

The accurate identification of root-knot nematode (RKN) species (Meloidogyne spp.) is essential for implementing management strategies. Methods based on the morphology of adults, isozymes phenotypes and DNA analysis can be used for the diagnosis of RKN. Traditionally, RKN species are identified by the analysis of the perineal patterns and esterase phenotypes. For both procedures, mature females are required. Over the last few decades, accurate and rapid molecular techniques have been validated for RKN diagnosis, including eggs, juveniles and adults as DNA sources. Here, we emphasized the methods used for diagnosis of RKN, including emerging molecular techniques, focusing on the major species reported in Brazil.(AU)

A identificação acurada de espécies do nematoide das galhas (NG) (Meloidogyne spp.) é essencial para a implementação de estratégias de manejo. Métodos baseados na morfologia de adultos, fenótipos de isoenzimas e análise de DNA podem ser usados para a diagnose do NG. Tradicionalmente, as espécies de NG são identificadas pela análise do padrão perineal e fenótipos de esterase. Em ambos os procedimentos, fêmeas maduras são necessárias. Nas últimas décadas, técnicas moleculares acuradas e rápidas têm sido validadas para a diagnose de NG, incluindo ovos, juvenis e adultos como fontes de DNA. Aqui, nós destacamos os métodos usados para a diagnose de NG, incluindo técnicas moleculares emergentes, focando nas principais espécies encontradas no Brasil.(AU)

Morphological, biochemical, and molecular characterization of Meloidogyne spp. populations from Brazilian soybean production regions / Caracterização morfológica, bioquímica e molecular de populações de Meloidogyne spp. de regiões brasileiras produtoras de soja

Soybean is a commodity of great economic importance worldwide, particularly in Brazil, worlds second largest producer. Nematodes, especially those of the Meloidogyne genus, severely limit productivity. Identification of nematode species is important for effective soybean management. Here, 26 populations of root-knot nematode (Meloidogyne spp.) from 15 municipalities in the states of Bahia, Mato Grosso, Goias, and Minas Gerais were characterized based on the morphology of the female perineal region, esterase profile, and identification based on amplification of specific regions of the population genome. Among the Meloidogyne spp. populations obtained, M. incognita and M. javanica, were identified. No mixed populations were present in the samples. Diagnosis based on molecular analysis was shown to be reliable and the fastest for characterization of nematode populations compared to other methods analyzed.

A soja é uma commodity de grande importância econômica em todo mundo, especialmente no Brasil, segundo maior produtor mundial. Os nematoides, em especial os do gênero Meloidogyne, causam grandes limitações na produtividade. A identificação das espécies de nematoides é uma informação importante para o manejo adequado. Neste trabalho, 26 populações do nematoide das galhas (Meloidogyne spp.), provenientes de municípios do estado da Bahia, Mato Grosso, Goiás e de Minas Gerais, foram caracterizados com base na morfologia da região perineal das fêmeas de Meloidogyne spp., seu perfil de esterase e identificação baseada em amplificação de regiões específicas do genoma dessas populações. Entre as populações de Meloidogyne spp. obtidas, identificou-se M. incognita e M. javanica. Não houve presença de populações mistas nas amostras analisadas. O diagnóstico baseado em análise molecular se mostrou mais rápido e confiável comparado ás outras análises.

Morphological, biochemical, and molecular characterization of Meloidogyne spp. populations from Brazilian soybean production regions / Caracterização morfológica, bioquímica e molecular de populações de Meloidogyne spp. de regiões brasileiras produtoras de soja

Soybean is a commodity of great economic importance worldwide, particularly in Brazil, worlds second largest producer. Nematodes, especially those of the Meloidogyne genus, severely limit productivity. Identification of nematode species is important for effective soybean management. Here, 26 populations of root-knot nematode (Meloidogyne spp.) from 15 municipalities in the states of Bahia, Mato Grosso, Goias, and Minas Gerais were characterized based on the morphology of the female perineal region, esterase profile, and identification based on amplification of specific regions of the population genome. Among the Meloidogyne spp. populations obtained, M. incognita and M. javanica, were identified. No mixed populations were present in the samples. Diagnosis based on molecular analysis was shown to be reliable and the fastest for characterization of nematode populations compared to other methods analyzed.(AU)

A soja é uma commodity de grande importância econômica em todo mundo, especialmente no Brasil, segundo maior produtor mundial. Os nematoides, em especial os do gênero Meloidogyne, causam grandes limitações na produtividade. A identificação das espécies de nematoides é uma informação importante para o manejo adequado. Neste trabalho, 26 populações do nematoide das galhas (Meloidogyne spp.), provenientes de municípios do estado da Bahia, Mato Grosso, Goiás e de Minas Gerais, foram caracterizados com base na morfologia da região perineal das fêmeas de Meloidogyne spp., seu perfil de esterase e identificação baseada em amplificação de regiões específicas do genoma dessas populações. Entre as populações de Meloidogyne spp. obtidas, identificou-se M. incognita e M. javanica. Não houve presença de populações mistas nas amostras analisadas. O diagnóstico baseado em análise molecular se mostrou mais rápido e confiável comparado ás outras análises.(AU)

Combining the essential oil of Piper aduncum L. with commercial insecticides / Combinação do óleo essencial de Piper aduncum L. com inseticidas comerciais

The use of synergists is important in minimizing the amount of chemical insecticide required for insect control. Their use can contribute to reducing environmental contamination and preserving beneficial insects. To further investigate a promising alternative to the synergist piperonyl butoxide (PBO), we compared the synergistic effects of PBO and Piper aduncum L. essential oil (PAEO) when combined with several insecticides (cypermethrin, zeta-cypermethrin, permethrin, and esfenvalerate) on the larvae of Spodoptera frugiperda (J. E. Smith, 1797). Initially, we determined the lethal doses and concentrations (LD50 and LC50) for S. frugiperda larvae subjected to separate treatments with PAEO and with each commercial insecticide. Subsequently, in order to evaluate the synergistic effect, combinations of sublethal doses or sublethal concentrations of the essential oil (½ and » of the LD50 or LC50, respectively) were prepared with sublethal doses or sublethal concentrations of the insecticides (below the LD40 or LC40). To confirm the evidence of the synergistic efficacy of the PAEO, the same reduced concentrations and doses of the insecticides that were previously used in combinations with the oil were also combined with PBO at a ratio of 10:1 (PBO:Insecticide). Through the relationship between the CL50 and DL50 of the insecticides taken separately and in their synergistic combinations with the PAEO and PBO, synergism factors (SF) were calculated for the various combinations. With residual contact, there was a significant enhancement of the commercial insecticides formulated with cypermethrin (SF = 73.03), zeta-cypermethrin (SF = 16.51), and permethrin (SF = 8.46-17.22) when combined with the PAEO; by contrast, with topical application there was a significant enhancement only for zeta-cypermethrin (SF = 0.40-4.26), permethrin (SF = 2.10-4.79), and esfenvalerate (SF = 3.80) when combined with the essential oil.[...]

A importância da utilização de sinergistas está relacionada à minimização da quantidade de inseticida químico necessária para o controle de insetos, podendo contribuir com a diminuição da contaminação ambiental e preservação de insetos benéficos. Na busca de uma alternativa ao butóxido de piperonila (PBO), o estudo comparou os efeitos do PBO e do óleo essencial de Piper aduncum L. combinados com os inseticidas cipermetrina, permetrina e esfenvalerato, quanto ao efeito sinérgico e homogeneidade de resposta de larvas de Spodoptera frugiperda (J. E. Smith, 1797). Inicialmente foram determinadas as doses e concentrações letais (DL50 e CL50) para larvas de S. frugiperda submetidas ao tratamento com o OEPA assim como para cada inseticida comercial considerados de forma isolada. Posteriormente, para avaliação do efeito sinérgico, foram realizadas combinações das doses e concentrações sub-letais com o óleo essencial (metade e um quarto da DL50 ou CL50, respectivamente) com as doses e concentrações sub-letais dos inseticidas comerciais (abaixo das DL40 ou CL40, respectivamente). Para complementar a comprovação da eficácia sinérgica do OEPA, foram utilizados como tratamentos adicionais as mesmas sub-concentrações e sub-doses dos inseticidas utilizadas anteriormente nas combinações com o óleo, passando a ser combinadas com o PBO na proporção de 10:1 (PBO: Inseticida). Por meio da relação das CL50 e DL50 dos inseticidas tomados isoladamente e de suas respectivas combinações sinérgicas com o OEPA e o PBO, foram obtidos os fatores de sinergismo (FS) para comparação entre si. Por contato residual foi evidenciada significativa potencialização dos inseticidas comerciais formulados com cipermetrina (FS= 73,03), zeta-cipermetrina (FS= 16,51) e permetrina (FS= 8,46-17,22), quando combinados com o OEPA.[...]

Combining the essential oil of Piper aduncum L. with commercial insecticides / Combinação do óleo essencial de Piper aduncum L. com inseticidas comerciais

The use of synergists is important in minimizing the amount of chemical insecticide required for insect control. Their use can contribute to reducing environmental contamination and preserving beneficial insects. To further investigate a promising alternative to the synergist piperonyl butoxide (PBO), we compared the synergistic effects of PBO and Piper aduncum L. essential oil (PAEO) when combined with several insecticides (cypermethrin, zeta-cypermethrin, permethrin, and esfenvalerate) on the larvae of Spodoptera frugiperda (J. E. Smith, 1797). Initially, we determined the lethal doses and concentrations (LD50 and LC50) for S. frugiperda larvae subjected to separate treatments with PAEO and with each commercial insecticide. Subsequently, in order to evaluate the synergistic effect, combinations of sublethal doses or sublethal concentrations of the essential oil (½ and » of the LD50 or LC50, respectively) were prepared with sublethal doses or sublethal concentrations of the insecticides (below the LD40 or LC40). To confirm the evidence of the synergistic efficacy of the PAEO, the same reduced concentrations and doses of the insecticides that were previously used in combinations with the oil were also combined with PBO at a ratio of 10:1 (PBO:Insecticide). Through the relationship between the CL50 and DL50 of the insecticides taken separately and in their synergistic combinations with the PAEO and PBO, synergism factors (SF) were calculated for the various combinations. With residual contact, there was a significant enhancement of the commercial insecticides formulated with cypermethrin (SF = 73.03), zeta-cypermethrin (SF = 16.51), and permethrin (SF = 8.46-17.22) when combined with the PAEO; by contrast, with topical application there was a significant enhancement only for zeta-cypermethrin (SF = 0.40-4.26), permethrin (SF = 2.10-4.79), and esfenvalerate (SF = 3.80) when combined with the essential oil.[...](AU)

A importância da utilização de sinergistas está relacionada à minimização da quantidade de inseticida químico necessária para o controle de insetos, podendo contribuir com a diminuição da contaminação ambiental e preservação de insetos benéficos. Na busca de uma alternativa ao butóxido de piperonila (PBO), o estudo comparou os efeitos do PBO e do óleo essencial de Piper aduncum L. combinados com os inseticidas cipermetrina, permetrina e esfenvalerato, quanto ao efeito sinérgico e homogeneidade de resposta de larvas de Spodoptera frugiperda (J. E. Smith, 1797). Inicialmente foram determinadas as doses e concentrações letais (DL50 e CL50) para larvas de S. frugiperda submetidas ao tratamento com o OEPA assim como para cada inseticida comercial considerados de forma isolada. Posteriormente, para avaliação do efeito sinérgico, foram realizadas combinações das doses e concentrações sub-letais com o óleo essencial (metade e um quarto da DL50 ou CL50, respectivamente) com as doses e concentrações sub-letais dos inseticidas comerciais (abaixo das DL40 ou CL40, respectivamente). Para complementar a comprovação da eficácia sinérgica do OEPA, foram utilizados como tratamentos adicionais as mesmas sub-concentrações e sub-doses dos inseticidas utilizadas anteriormente nas combinações com o óleo, passando a ser combinadas com o PBO na proporção de 10:1 (PBO: Inseticida). Por meio da relação das CL50 e DL50 dos inseticidas tomados isoladamente e de suas respectivas combinações sinérgicas com o OEPA e o PBO, foram obtidos os fatores de sinergismo (FS) para comparação entre si. Por contato residual foi evidenciada significativa potencialização dos inseticidas comerciais formulados com cipermetrina (FS= 73,03), zeta-cipermetrina (FS= 16,51) e permetrina (FS= 8,46-17,22), quando combinados com o OEPA.[...](AU)

Detection of carboxylesterase and esterase activity in culturable gut bacterial flora isolated from diamondback moth, Plutella xylostella (Linnaeus), from India and its possible role in indoxacarb degradation

Diamondback moth (DBM), Plutella xylostella (Linnaeus), is a notorious pest of brassica crops worldwide and is resistant to all groups of insecticides. The insect system harbors diverse groups of microbiota, which in turn helps in enzymatic degradation of xenobiotic-like insecticides. The present study aimed to determine the diversity of gut microflora in DBM, quantify esterase activity and elucidate their possible role in degradation of indoxacarb. We screened 11 geographic populations of DBM in India and analyzed them for bacterial diversity. The culturable gut bacterial flora underwent molecular characterization with 16S rRNA. We obtained 25 bacterial isolates from larvae (n = 13) and adults (n = 12) of DBM. In larval gut isolates, gammaproteobacteria was the most abundant (76%), followed by bacilli (15.4%). Molecular characterization placed adult gut bacterial strains into three major classes based on abundance: gammaproteobacteria (66%), bacilli (16.7%) and flavobacteria (16.7%). Esterase activity from 19 gut bacterial isolates ranged from 0.072 to 2.32 µmol/min/mg protein. Esterase bands were observed in 15 bacterial strains and the banding pattern differed in Bacillus cereus KC985225 and Pantoea agglomerans KC985229. The bands were characterized as carboxylesterase with profenofos used as an inhibitor. Minimal media study showed that B. cereus degraded indoxacarb up to 20%, so it could use indoxacarb for metabolism and growth. Furthermore, esterase activity was greater with minimal media than control media: 1.87 versus 0.26 µmol/min/mg protein. Apart from the insect esterases, bacterial carboxylesterase may aid in the degradation of insecticides in DBM.(AU)

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 a-and b-esterase patterns of four economically important table-wine grape cultivars (Italia, Rubi, Benitaka and Brasil) of Vitis vinifera. The a- and b-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 a- and b-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.(AU)

O presente estudo caracterizou o efeito de diferentes fungicidas comumente aplicados como medidas de controle de pragas sobre padrões de - e -esterases de quatro importantes cultivares de uva de mesa (Itália, Rubi, Benitaka e Brasil) de Vitis vinifera. Os padrões de - e -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 - e -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.(AU)

Caracterização de Malassezia spp isoladas de conduto auditivo de cães e gatos e avaliação dos fatores virulência

O gênero Malassezia compreende 17 espécies de leveduras lipofílicas e lipodependentes, componentes naturais da microbiota da pele dos animais homeotérmicos, incluindo humanos, e conhecido agente oportunista em dermatites e otites. A maior parte da casuística de otite fúngica em cães é atribuída a M. pachydermatis, mas espécies lipodependentes também podem ser agentes desta afecção, sendo possível que haja subdiagnósticos. Na rotina clínica veterinária, é comum o uso do exame direto de amostras otológicas como ferramenta auxiliar para direcionar uma melhor conduta terapêutica. Tendo esses pontos em vista, o trabalho objetiva realizar isolamento de Malassezia spp. a partir de amostras de otite de cães e gatos, identificando-as à nível de espécie, observar a presença da Malassezia em infeções mistas ou puras, verificar in vitro a produção dos principais fatores de virulência inerentes à patogenicidade e correlacionar a produção dos fatores de virulência com a natureza da infecção. Foram utilizadas 170 amostras oriundas dos condutos auditivos externos de 94 cães e 11 gatos com otite clínica, encaminhadas ao Laboratório de Leveduras Patogênicas e Ambientais, onde foram semeadas nos meios Agar Dixon Modificado e Agar Sabouraud Dextrose 2% com Cloranfenicol. Foram realizados testes clássicos para identificação fenotípica, como preconizado por Kurtzman et al. (2011) e análise proteômica por Maldi-TOF e bioquímico por VITEK2 ® para confirmação da identificação. Para análise da virulência foram testadas as produções de proteases, fosfolipase, lipases, esterases e hemolisina. Dos 105 animais, 63% (66) animais possuiam Malassezia sp. nas suas amostras, sendo uma lipodependente, e 37% (39) não tinham a levedura. O valor preditivo positivo e negativo do exame direto foram 75,58 e 67,19%, respectivamente. A sensibilidade do exame em relação ao isolamento foi de 75,58% e a especificidade de 67,19%. Não houve diferença estatística entre a recuperação das leveduras no diagnóstico em Agar Sabouraud Dextrose 2% e Agar Dixon Modificado. A identificação por espetrofotometria de massa teve 82% de acurácia na identificação dos isolados e as restantes não tiveram scores de identificação, enquanto o VITEK ® , identificou erroneamente todas as amostras. Para os testes enzimáticos, 100% (96) das amostras foram positivas para a produção de proteases e lipases. Apenas uma amostra não foi produtora de fosfolipase e esterase. 89% (86) amostras foram positivas para a produção de hemolisina. Conclui-se que a utilização do Agar Dixon Modificado é importante na rotina laboratorial de diagnóstico microbiologico de otites canina e felina, para ampliar a gama de espécies de Malassezia diagnosticadas. O exame direto mostrou ser um bom indicador para a presença da levedura, mas não necessáriamente para indicar seu envolvimento na infecção. Para além, percebe-se que essas leveduras in vitro possuem alta atividade enzimática, podendo fazer parte da patogênese da otite microbiana, mesmo quando não é o principal agente envolvido, como nas otites mistas, mesmo as que possuem contagens baixas no isolamento.

The genus Malassezia currently comprises 17 species of lipophilic and lipid- dependent yeasts. Considered a normal part of the skin microbiota of warm-blooded vertebrates, including humans, these may also become opportunistic agents in dermatitis and otitis. Fungal otitis in dogs have been associated with M. pachydermatis, but lipid- dependent species are also commonly found, indicating a possibility of frequent misdiagnosis. In the veterinary clinical routine, microscopy of otological samples is used as an auxiliary tool towards the implementation of an adequate drug therapy. The aim of this work was the isolation of Malassezia spp. from otitis samples from dogs and cats, their respective identification at the species level, observe the presence of Malassezia in pure or co-infections, in vitro detection the main virulence factors inherent to the genus pathogenicity being produced, and the establishment of a correlation between the detected virulence factors being produced and the nature of the infection. A total of 170 samples from the external ear canal of 94 dogs and 11 cats with clinical otitis were sent to the Laboratory of Pathogenic and Environmental Yeasts, where they were inoculated in Modified Dixon Agar and Sabouraud Dextrose 2% Agar supplemented with chloramphenicol. Classical methods allowed for the phenotypic identification, as recommended by Kurtzman et al. (2011) further corroborated through proteomic analysis by MALDI-TOF MS and biochemical analysis by VITEK2 ® . For virulence analysis, protease, phospholipase, lipase, esterase and hemolysin production were tested. Of the 105 animals, 63% (66) individuals were positive for Malassezia sp., as one lipid dependent, and 37% (39) were negative for the genus. The positive and negative predictive values of the direct exam were 75.58% and 67.19%, respectively. The sensitivity of the isolation test was 75.58% and the specificity 67.19%. There was no significant difference between yeast recovery in Sabouraud Dextrose 2% Agar and Modified Dixon Agar. The identification by mass spectrophotometry had an 82% accuracy in the identification of the isolates and remaining isolates did not reach the identification score, whereas VITEK® misidentify all the samples. For the enzymatic tests, 100% (96) of the samples were positive for the production of proteases and lipases. Only one sample did not produce phospholipase nor esterase. 89% (86) samples were hemolysin producers. It was possible to conclude that the use of Modified Dixon Agar is important in the laboratory routine for microbiological diagnosis of canine and feline otitis, extending the range of Malassezia species diagnosed. Direct examination proved to be a good indicator of the presence of yeast, but not enlightening regarding its involvement in the infection. In addition, these yeasts have high enzymatic activity in vitro and may be part of the pathogenesis of microbial otitis, even when it is not the main pathological agent, as in co-infections otitis even with low isolation counts. Key words: malasseziosis, microbiological diagnosis, fungal infection

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.(AU)

Comparative electrophoretic profile of proteins and esterases in healthy silkworm larvae (Bombyx mori Lineu, 1758) and infected with nucleopolyhedrovirus / Perfil eletroforético comparativo de proteínas e esterases em lagartas de bicho da seda (Bombyx mori Lineu, 1758) saudáveis e infectadas por nucleopoliedrovírus

Total proteins and esterases from silk gland extracts of Bombyx mori silkworm were characterized and compared with electrophoretic profiles of prepared extracts with silkworm glands infected with nucleopolyhedrovirus (BmNPV). SDS-PAGE (7%) gels was used for the total proteins, and 10% native PAGE for esterases. In the silk glands extracts of healthy silkworms, it was observed seven protein zones with molecular weight varying between 10 kDa (P1) and 60 kDa (P7). In the infected silkworms, a new zone named P8 (90 kDa) was also detected. Esterases activity at 5th instar larvae underwent changes after the infection with BmNPV, since there was a reduction (EST-6 and EST-7) and an increase (EST-8) in the intensity of the regions of esterases activity, and specificity of EST-9 to -naphthyl acetate. Those alterations observed in the expression of genes after the infection with the nucleoplyhedrovirus can be used as markers to detect infections in B. mori.(AU)

Nesse estudo foram identificadas e comparadas as alteraçُes das proteيnas totais e esterases em extratos de glândula sericيgena de lagartas de Bombyx mori sadias e infectadas por nucleopoliedrovيrus (BmNPV), empregando eletroforese SDS-PAGE com géis a 7% para proteيnas totais, e PAGE a 10% para esterases. Nos extratos de glândulas sericيgenas de lagartas saudلveis, foram observadas sete regiُes proteicas com peso molecular que variam entre 10 kDa (P1) a 60 kDa (P7). Naquelas infectadas pelo BmNPV, uma nova regiمo denominada de P8 (90 kDa) foi detectada. A atividade das esterases no 5؛ instar larval sofreu alteraçُes apَs a infecçمo pelo BmNPV porque houve reduçمo (EST-6 e EST-7) e aumento (EST-8) na intensidade das regiُes de atividade de esterases; e especificidade da EST-9 com o substrato -naftil acetato. Essas alteraçُes na expressمo gênica, apَs a infecçمo pelo nucleopoliedrovيrus, poderمo ser utilizadas como marcadores para detecçمo da infecçمo em B. mori.(AU)

Genetic Analysis of Izoenzymes Polymorphisms in Silkworm (Bombyx mori L. ) Strains / Análise genética de polimorfismos bioquímicos em linhagens do bicho da seda (Bombyx mori L. )

This work carried out to evaluates the polymorhism in the silkworm of different lineages using the isoenzymes electrophoresis to detect biochemical markers and to investigate the genetics of populations for those lineages. They were used as samples individual extracts of silk glands of second day old larvas of the fifth instar, originating from seven Japanese lineages and eight pure Chinese lineages maintained by the Cocamar-Cooperativa Agroindustrial de Maringل. The isozymes acid phosphatase (ACP), alkaline phosphatase (AKP) and carbonic anhydrase (CA) they were submitted to the electrophoresis in starch gels 14%. The esterases (EST) were analyzed in polyacrylamide gels to 10% and stained with and â-naphtyl acetate. The total of 21 loci was detected, and 04 (19.05%) they are polymorphic, Est-11, Acp, Akp, Ca. The fixation index (Fis) for the analyzed isozymes it was 0.0751, indicating excess of homozygotes. The value of Fst (0.6128) it shows that the lineages are well differentiated. The dendrogram obtained with the values of genetic distance didn"t separate the Chinese and Japanese lineages analyzed totally. That preliminary evaluation of the lineages of B. mori shows that they present genetic material that it can be used in breeding programs that have the purpose of producing hybrid for silk production.(AU)

Este trabalho teve como objetivo avaliar o polimorfismo em lagartas do bicho-da-seda de diferentes linhagens utilizando a eletroforese de isoenzimas para detectar marcadores bioquيmicos e investigar a genética de populaçُes para essas linhagens. Foram utilizados como amostras extratos individuais de glândulas sericيgenas de lagartas do segundo dia da quinta idade, de sete linhagens japonesas e oito linhagens chinesas puras mantidas pela Cocamar-Cooperativa Agroindustrial de Maringل. As isozimas fosfatase لcida (ACP), fosfatase alcalina (AKP) e anidrase carbônica (CA) foram avaliadas por meio de eletroforese em géis de amido de milho a 14%. As esterases (EST) foram analisadas por meio de eletroforese vertical em géis de poliacrilamida a 10% e coloraçمo com ل e â-naftil acetato. Foram observados 21 locos, dentre os quais quatro (19.05%) sمo polimَrficos, Est-11, Acp, Akp, Ca. O يndice de fixaçمo (Fis) para as isozimas analisadas foi 0.0751, indicando excesso de homozigotos. O valor de Fst (0.6128) permite sugerir que as linhagens estمo bem diferenciadas. O dendograma obtido a partir dos valores de distância genética nمo separou totalmente as linhagens chinesas e japonesas analisadas. Essa avaliaçمo preliminar das linhagens de B. mori mostra que elas apresentam material genético que pode ser utilizado em programas de cruzamentos que tenham a finalidade de produzir hيbridos para prara produção de seda.(AU)

Variability in the production of extracellular enzymes by entomopathogenic fungi grown on different substrates

Entomopathogenic fungi are important controllers of pest-insects populations in agricultural production systems and in natural environment. These fungi have enzymatic machinery which involve since the recognition and adherence of spores in their hosts culminating with infection and death of these insects. The main objective of this study was to analyzed extracellular enzyme production of the fungi strains Beauveria bassiana, Metarhizium anisopliae and Paecilomyces sp when cultured on substrates. These fungi were grown in minimal media containing specific substrates for the analysis of different enzymes such as amylases, cellulases, esterases, lipases, proteases (gelatin and caseinase), pectinases and cuticles of Musca domestica larvae and adults. All the assays were performed with and without the presence of dextrose in the culture media. The quantification of enzyme activity was performed by the ratio of halo / colony (H/C) and the results subjected to variance analysis level of 5% (ANOVA) followed by post-Tukey test. All strains were positive for lipase and also they showed a high significant enzyme production for gelatin at concentrations of 4 and 1%. B. bassiana and Paecilomyces sp. were positive for amylase, pectinase and caseinase, and only Paecilomyces sp. showed cellulase activity.