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)

Accelerated ageing test and enzymatic expression in the evaluation of kale seed quality / Teste de envelhecimento acelerado e expressão enzimática na avaliação da qualidade de sementes de couve

The demand for high-quality vegetable seeds and the production of vigorous seedlings has increased in recent years, as these characteristics are determining factors of production success. Vegetables are growing in national importance, and kale stands out as an important source of income for small farmers. The objective of this study was to adapt the traditional accelerated aging test methodology with a saturated NaCl solution of kale seeds and evaluate the enzymatic activity after the vigour test. Six batches of kale seeds were used, and the moisture content, weight of one thousand seeds, first germination count, germination, germination speed index, emergence, initial stand, and emergence velocity index were determined. In the accelerated ageing test, the seeds were submitted to the traditional accelerated ageing test method and the accelerated ageing test method with a saturated NaCl solution for ageing periods of 0, 24, 48, 72, and 96 hours. Electrophoretic analysis of superoxide dismutase (SOD), catalase (CAT), and alcohol dehydrogenase (ADH) isoenzymes was also performed. The accelerated ageing test, using the traditional method for 72 hours at 41 °C, is adequate for evaluating the physiological potential of kale seeds. The isoenzymatic analyses of SOD, CAT, and ADH demonstrate that the biochemical markers are efficient at differentiating kale seeds after accelerated ageing.(AU)

A demanda por sementes de hortaliças com alta qualidade e a obtenção de mudas vigorosas tem aumentado nos últimos anos, pois estas características constituem fatores determinantes do êxito da produção. As hortaliças têm uma importância nacional crescente, dentre elas se destaca a couve, por ser importante fonte de renda para pequenos agricultores. Objetivou-se adequar a metodologia do teste de envelhecimento acelerado tradicional e com solução saturada de NaCl em sementes de couve e avaliar a atividade enzimática após o teste de vigor. Foram utilizados seis lotes de sementes de couve e determinouse o grau de umidade, peso de mil sementes, primeira contagem de germinação, germinação, índice de velocidade de germinação, emergência, estande inicial e índice de velocidade de emergência. No teste de envelhecimento acelerado, as sementes foram submetidas ao método tradicional e com solução saturada de NaCl, por períodos de 0; 24; 48; 72 e 96 horas. Realizou-se também a análise eletroforética das isoenzimas superóxido dismutase (SOD), catalase (CAT) e álcool desidrogenase (ADH). O teste de envelhecimento acelerado, utilizando o método tradicional na combinação de 72 horas a 41 °C é adequado para avaliar o potencial fisiológico de sementes de couve. As análises isoenzimáticas superóxido dismutase (SOD), catalase (CAT) e álcool desidrogenase (ADH) demonstram que os marcadores bioquímicos são eficientes e promissores na diferenciação de sementes de couve após o envelhecimento acelerado.(AU)

Changes in serum adenosine deaminase and isoenzyme levels in addition to routine liver biochemical parameters in sheep with chronic fascioliasis / Alterações nos níveis séricos de adenosina desaminase e isoenzima em relação aos parâmetros bioquímicos de rotina do fígado de ovinos com fasciolíase crônica

This study assessed changes in the levels of adenosine deaminase (ADA) and its isoenzymes in addition to routine liver biochemical parameters in sheep with fascioliasis. The study was conducted on 35 Akkaraman sheep. Of these, 25 sheep were diagnosed with fascioliasis based on anamnesis and clinical signs, and had endoparasites based on parasitological examinations (Fasciola-infected group). The remaining 10 sheep that were sampled from a single healthy herd (same flock) different from the infected group did not have any clinical signs or endoparasites (control group). Total protein (TP), albumin (ALB), and globulin (GLB) levels gradually increased on days after treatment compared to the values measured before treatment; the increases were statistically significant on all days for TP levels but only on day 14 after treatment for GLB levels (P < 0.05). Although, the ALB levels did not increase significantly on days after treatment, the ALB level and ALB/GLB ratio on days 7 and 14 after treatment were still lower than the values of day 21 after treatment and control group (P < 0.05). Total bilirubin (T-Bil) and direct bilirubin (D-Bil) levels on days 14 and 21 were significantly lower than that of day 0 (before treatment) and day 7 after treatment (P < 0.05). These results indicated that the increase in adenosine deaminase (ADA) and ADA1 levels may be due to possible concomitant infection of Fasciola larvae (in the parenchyma) and adults (in the bile duct).

Este estudo teve como objetivo avaliar as alterações nos níveis de adenosina desaminase (ADA) e suas isoenzimas, além de parâmetros bioquímicos hepáticos de rotina em ovinos com fasciolíase. O estudo foi realizado em 35 ovinos s Akkaraman. Destes, 25 ovinos foram suspeitas de ter fasciolíase com base na anamnese e sinais clínicos, e diagnosticados com endoparasitas com base em exames parasitológicos (grupo de estudo). Os 10 ovinos restantes que foram amostrados no mesmo rebanho não apresentavam quaisquer sinais clínicos ou endoparasitas (grupo controle). Os níveis de proteína total (TP), albumina (ALB) e globulina (GLB) aumentaram gradualmente nos dias após o tratamento em comparação com os valores antes do tratamento; os aumentos foram estatisticamente significativos em todos os dias para os níveis de TP, mas apenas após o dia 14 de tratamento para os níveis de GLB (P < 0,05). Embora os níveis de ALB não tenham aumentado significativamente após os dias de tratamento, o nível de ALB e a razão ALB / GLB após o tratamento nos dias 7 e 14 ainda eram menores do que após o tratamento no dia 21 e os valores do grupo de controle (P < 0,05). Os níveis de bilirrubina total (T-Bil) e bilirrubina direta (D-Bil) nos dias 14 e 21 após o tratamento foram significativamente menores do que os valores do dia 7 antes do tratamento e após o tratamento (P < 0,05). Estes resultados indicam que o aumento nos níveis de adenosina desaminase (ADA) e ADA1 pode ser devido à possível infecção concomitante de larvas de Fasciola (no parênquima) e adultos (nos dutos).

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)

Tratamento da epilepsia idiopática canina - quadro clínico após interação medicamentosa / Treatment of canine idiopathic epilepsy - clinical picture after drug interaction

Background: The large number of diseases demand perennial development of the pharmaceutical industry. The drugtesting phase is essential to make them available safely. Awareness of pharmacological properties, adverse effects and drug interactions is required. Drug interactions are common in veterinary medicine and should be avoided. At times, epileptic seizures require polydrug therapy, predisposing patients to drug interactions. The interaction between carbamazepine and phenobarbital reported in the literature is an example. The aim of this paper is to report a clinical picture of drug interaction in the treatment of idiopathic epilepsy. Case: A 1-year-old Border Collie male dog, was admitted at the Veterinary Hospital of the Federal University of Lavras in post-ictal. The tutor reported that a year ago the animal had epileptic seizures and clusters with intervals of 21 to 25 days. Despite the continued use of previously prescribed phenobarbital (7.4 mg/kg, v.o., BID, until new recommendations) and carbamazepine (7.5 mg/kg, v.o., BID, until new recommendations), seizure control was not achieved. The physical examination indicated, tachypnea, ptialism, mydriasis, intense fatigue, and alienation from the environment. The patient did not respond to the threat-reflex test. Blood count, hepatic and renal blood chemistry, serum electrolyte (potassium, sodium, calcium and phosphorus), and phenobarbital dosages were requested. Based on the animal's history, breed characteristics, and alterations in the physical examination associated with normal results in complementary exams, idiopathic epilepsy was diagnosed. After analyzing the case, it was observed that the inefficiency in the control of seizures was possibly due to the drug interaction between phenobarbital and carbamazepine. Carbamazepine and phenobarbital reciprocally reduce their half-lives. To confirm the raised hypothesis, the serum concentration of carbamazepine was gradually reduced through weaning from its dose administered to the patient. Serial dosage of the concentration of phenobarbital in the bloodstream was performed. As a result, the serum phenobarbital, previously dosed at a concentration of 13.3 mg/dL with concomitant administration of carbamazepine, increased to 22 mg/dL 40 days after the beginning of weaning from carbamazepine (T0), and then to 36 mg/dL 100 days after T0. There was an increase in the concentration of phenobarbital in the bloodstream while the serum concentration of carbamazepine declined. The patient spaced out his seizures to every 50 to 60 days with phenobarbital monotherapy at a dose of 6 mg/kg. Discussion: Efficient control of clusters, such as the reduction of seizures by 50%, was only possible due to the meticulous perception of the possible interaction reported in medicine. Carbamazepine and phenobarbital are P450 isoenzyme inducers. The concomitant administration of both drugs potentiated the action of isoenzymes in the hepatic microsomal system, which led to an accelerated metabolic processing of the drugs. After weaning from carbamazepine, that is, reducing the action of carbamazepine on the isoenzymes of the P450 enzyme system, the concentration of phenobarbital normalized at 36 mg/ dL. Such concentration is within the reference range reported in the literature: 25 mg/dL to 35 mg/dL of serum phenobarbital for treatment efficacy. Therefore, the control of convulsive crises was achieved. The increase in the concentration of phenobarbital due only to weaning from carbamazepine, even after decreasing the daily dose of barbiturate prescribed to the animal, contributed to evidence of the interaction of these drugs. It is noted that prior knowledge of pharmacological properties, careful study of the patient's history, and the cooperation of the tutor were essential for the therapeutic success and practice of evidence-based veterinary medicine.

Population structure and phylogenetic relationships in Brassica rapa L. subspecies by using isozyme markers / Estrutura populacional e relações filogenéticas em subespécies de Brassica rapa L. utilizando marcadores isoenzimáticos

The present study aimed to assess population structure and phylogenetic relationships of nine subspecies of Brassica rapa L. represented with thirty-five accessions cover a wide range of species distribution area using isozyme analysis in order to select more diverse accessions as supplementary resources that can be utilized for improvement of B. napus. Enzyme analysis resulted in detecting 14 putative polymorphic loci with 27 alleles. Mean allele frequency 0.04 (rare alleles) was observed in Cat4A and Cat4B in sub species Oleifera accession CR 2204/79 and in subspecies trilocularis accessions CR 2215/88 and CR 2244/88. The highest genetic diversity measures were observed in subspecies dichotoma, accession CR 1585/96 (the highest average of observed (H0) and expected heterozygosity (He), and number of alleles per locus (Ae)). These observations make this accession valuable genetic resource to be included in breeding programs for the improvement of oilseed B. napus. The average fixation index (F) is significantly higher than zero for the analysis accessions indicating a significant deficiency of heteozygosity. The divergence among subspecies indicated very great genetic differentiation (FST = 0.8972) which means that about 90% of genetic diversity is distributed among subspecies, while 10% of the diversity is distributed within subspecies. This coincides with low value of gene flow (Nm = 0.0287). B. rapa ssp. oleifera (turnip rape) and B. rapa ssp. trilocularis (sarson) were grouped under one cluster which coincides with the morphological classification.(AU)

O presente estudo teve como objetivo avaliar a estrutura populacional e as relações filogenéticas de nove subespécies de Brassica rapa L. representadas com 35 acessos, cobrindo uma ampla gama de áreas de distribuição de espécies usando análise isoenzimática, a fim de selecionar acessos mais diversos como recursos suplementares que podem ser utilizados para melhoria de B. napus. A análise enzimática resultou na detecção de 14 loci polimórficos putativos com 27 alelos. A frequência média de 0,04 alelo (alelos raros) foi observada em Cat4A e Cat4B, nas subespécies Oleifera CR 2204/79 e nas subespécies trilocularis CR 2215/88 e CR 2244/88. As maiores medidas de diversidade genética foram observadas na subespécie dicotômica CR 1585/96 (a média mais alta observada (H0) e heterozigosidade esperada (He) e número de alelos por locus (Ae). Essas observações tornam esse acesso um valioso recurso genético a ser incluído em programas de melhoramento de oleaginosas B. napus. O índice médio de fixação (F) é significativamente maior que 0 para os acessos à análise, indicando uma deficiência significativa de heterozigose. A divergência entre as subespécies indicou uma grande diferenciação genética (FST = 0,8972), o que significa que cerca de 90% da diversidade genética é distribuída entre as subespécies, enquanto 10% da diversidade é distribuída nas subespécies. Isso coincide com o baixo valor do fluxo gênico (Nm = 0,0287). B. rapa ssp. oleifera (nabo) e B. rapa ssp. trilocularis (sarson) foram agrupados conforme a classificação morfológica.(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.

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)

Supplanting resistance of the Mi gene by root-knot nematode in industrial tomato in the Cerrado in Goiás State of Brazil / Suplantação de resistência do gene Mi por nematoide-das-galhas em tomate industrial no Cerrado de Goiás, Brasil

Problems with root-knot nematodes, caused by genus Meloidogyne, can be controlled through the introduction of resistance genes in commercial varieties of tomatoes. However, plants of the hybrid ‘Heinz 9992 for industrial processing, carrying the Mi gene, were reported with their roots severely infected in experimental cultivation in Goiás State of Brazil with symptoms typical of galls caused by Meloidogyne spp. After dissecting the galls present in the root samples, the females of the nematodes were removed and afterwards, performed the analysis of the perineal pattern and the biochemical characterization by electrophoresis of isoenzymes, besides the pathogenicity test. The pathogen analyzed was identified as Meloidogyne incognita. The pathogenicity test confirmed the supplanting resistance of the tomato hybrid with the Mi gene by this virulent population of M. incognita.(AU)

Problemas causados pelos nematoides-das-galhas, pertencentes ao gênero Meloidogyne podem ser controlados com a introdução de genes de resistência em cultivares comerciais de tomateiro. No entanto, plantas do híbrido ‘Heinz 9992 adequadas ao processamento industrial, portadoras do gene Mi, foram encontradas com suas raízes severamente infectadas, em um cultivo experimental no município de Morrinhos (GO) com sintomas típicos de galhas causadas por Meloidogyne spp. Após dissecar as galhas presentes nas amostras de raízes, as fêmeas do nematoide foram retiradas para a análise do padrão perineal e a caracterização bioquímica por eletroforese de isoenzimas, além do teste de patogenicidade. O patógeno em estudo foi identificado como Meloidogyne incognita. O teste de patogenicidade confirmou que ocorreu a suplantação de resistência do híbrido de tomateiro, portador do gene Mi, por esta população virulenta de M. incognita.(AU)

Aplicação de silício no solo: efeito na expressão enzimática de sementes oriundas de plantas de arroz sob estresse salino / Application of silicon to the soil: effect on enzymatic expression in rice seed produced under salt stress

A utilização de silício pode acarretar em aumento da capacidade biológica da planta em resistir às condições adversas do meio ambiente. Diante disso, o presente trabalho teve por objetivo avaliar a expressão isoenzimática de sementes oriundas de plantas cultivadas com aplicação de cinza da casca de arroz, via solo, sob estresse salino. O delineamento utilizado foi em blocos casualizados, em esquema fatorial A x B (Fator A- Cinco doses de cinza da casca de arroz aplicada via solo: 0; 500; 1000; 1500 e 2000 kg ha-¹; Fator B- Três concentrações salinas: 0, 4 e 8 mM), com quatro repetições. Foram realizados os testes de primeira contagem de germinação e germinação, e expressão dos sistemas isoenzimáticos glutamato oxaloacetato transaminase, álcool desidrogenase, glutamato desidrogenase e sorbitol. Variações nos quatro sistemas eletroforéticos analisados podem estar associados ao estresse salino causado durante o crescimento e desenvolvimento de plantas de arroz. O sistema isoenzimático glutamato oxaloacetato transaminase é uma ferramenta complementar à avaliação do potencial fisiológico de sementes de arroz.

The silicon utilization may result in the increased biological ability of the plants to withstand the harsh conditions of the environment. In this sense, the study aimed to evaluate the effect of the application of rice husk ash to the soil in rice plants under salt stress on isoenzymes expression. The experimental design was a randomized block in factorial A x B (Factor A- Five doses of rice husk ash applied to soil: 0.0, 500, 1000, 1500, and 2000 kg ha-¹, Factor B- Three salt concentrations: 0, 4, and 8 mM), with four replications. Tests were the first count of germination and germination, and expression of isozyme systems glutamate oxaloacetate transaminase, alcohol dehydrogenase, glutamate dehydrogenase, and sorbitol. Variations in the all analyzed electrophoretic systems may be associated with salt stress caused during the growth and development of rice plants. The isoenzyme system glutamate oxaloacetate transaminase is a complementary tool to assess the physiological rice seed quality.

Aplicação de silício no solo: efeito na expressão enzimática de sementes oriundas de plantas de arroz sob estresse salino / Application of silicon to the soil: effect on enzymatic expression in rice seed produced under salt stress

A utilização de silício pode acarretar em aumento da capacidade biológica da planta em resistir às condições adversas do meio ambiente. Diante disso, o presente trabalho teve por objetivo avaliar a expressão isoenzimática de sementes oriundas de plantas cultivadas com aplicação de cinza da casca de arroz, via solo, sob estresse salino. O delineamento utilizado foi em blocos casualizados, em esquema fatorial A x B (Fator A- Cinco doses de cinza da casca de arroz aplicada via solo: 0; 500; 1000; 1500 e 2000 kg ha-¹; Fator B- Três concentrações salinas: 0, 4 e 8 mM), com quatro repetições. Foram realizados os testes de primeira contagem de germinação e germinação, e expressão dos sistemas isoenzimáticos glutamato oxaloacetato transaminase, álcool desidrogenase, glutamato desidrogenase e sorbitol. Variações nos quatro sistemas eletroforéticos analisados podem estar associados ao estresse salino causado durante o crescimento e desenvolvimento de plantas de arroz. O sistema isoenzimático glutamato oxaloacetato transaminase é uma ferramenta complementar à avaliação do potencial fisiológico de sementes de arroz.(AU)

The silicon utilization may result in the increased biological ability of the plants to withstand the harsh conditions of the environment. In this sense, the study aimed to evaluate the effect of the application of rice husk ash to the soil in rice plants under salt stress on isoenzymes expression. The experimental design was a randomized block in factorial A x B (Factor A- Five doses of rice husk ash applied to soil: 0.0, 500, 1000, 1500, and 2000 kg ha-¹, Factor B- Three salt concentrations: 0, 4, and 8 mM), with four replications. Tests were the first count of germination and germination, and expression of isozyme systems glutamate oxaloacetate transaminase, alcohol dehydrogenase, glutamate dehydrogenase, and sorbitol. Variations in the all analyzed electrophoretic systems may be associated with salt stress caused during the growth and development of rice plants. The isoenzyme system glutamate oxaloacetate transaminase is a complementary tool to assess the physiological rice seed quality.(AU)

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)

New human case reports of cutaneous leishmaniasis by Leishmania (Viannia) naiffi in the Amazon region, Brazil / Novos relatos de casos de leishmaniose cutânea por Leishmania (Viannia) naiffi na região amazônica, Brasil

Few cases of human cutaneous leishmaniasis (CL) caused by Leishmania naiffi were described in the medical literature. The aim of this study was to report and analyze new cases of L. naiffi in the period between the years 1992 to 2011. The strains were characterized by isoenzyme analysis. All patients assisted had small lesions; ranging from 1.0x1.0 mm and 13.5x11.5 mm. The lesions observed were widely distributed: 55.5% on the lower limb, 5.5% in the abdominal area, 16.6% on upper limb and 22.2% in upper limb and back. Seventy-two percent of patients had ulcerated lesions. Clinical course of the disease varied from 1 to 10 months. According to gender, most infected individuals were men (83.3%). The patients came from Amazonas (10), Pará (01) and Rondônia (01), north States of Brazil. Five patients were infected in experimental stations of the National Institute of Amazonian Research (INPA). Although the results of this study were similar to other reports in the literature, some of the patients had more of one skin lesion. It is also reported the first case of CL caused by L. naiffi in the State of Rondônia and identified an area of disease transmission in the experimental station of INPA.(AU)

Poucos casos de leishmaniose cutânea humana (LC) causada por Leishmania naiffi foram descritos na literatura médica. Assim, o objetivo deste estudo foi relatar e analisar novos casos de infecção por L. naiffi no período compreendido entre os anos de 1992 a 2011. As cepas foram caracterizadas por análise de isoenzimas. Todos os pacientes atendidos apresentavam lesões pequenas; variando entre 1,0x1,0 mm e 13,5x11,5 mm. As lesões observadas foram amplamente distribuídas: 55,5% no membro inferior, 5,5% na área abdominal, 16,6% no membro superior e 22,2% no membro superior e para trás. Setenta e dois por cento dos pacientes tiveram lesões ulceradas. O curso clínico da doença variou de 1 a 10 meses. De acordo com o sexo, a maioria dos indivíduos infectados eram homens (83,3%). Os pacientes vieram de Amazonas (10), Pará (01) e Rondônia (01), estados do norte do Brasil. Cinco pacientes foram infectados em estações experimentais do Instituto Nacional de Pesquisas da Amazônia (INPA). Embora os resultados encontrados neste estudo fossem semelhantes às outras descrições na literatura, alguns dos pacientes apresentavam mais de uma lesão cutânea. Também é relatado o primeiro caso de LC causada por L. naiffi no Estado de Rondônia e identificada uma área de transmissão da doença na estação experimental do INPA.(AU)

New human case reports of cutaneous leishmaniasis by Leishmania (Viannia) naiffi in the Amazon region, Brazil / Novos relatos de casos de leishmaniose cutânea por Leishmania (Viannia) naiffi na região amazônica, Brasil

Few cases of human cutaneous leishmaniasis (CL) caused by Leishmania naiffi were described in the medical literature. The aim of this study was to report and analyze new cases of L. naiffi in the period between the years 1992 to 2011. The strains were characterized by isoenzyme analysis. All patients assisted had small lesions; ranging from 1.0x1.0 mm and 13.5x11.5 mm. The lesions observed were widely distributed: 55.5% on the lower limb, 5.5% in the abdominal area, 16.6% on upper limb and 22.2% in upper limb and back. Seventy-two percent of patients had ulcerated lesions. Clinical course of the disease varied from 1 to 10 months. According to gender, most infected individuals were men (83.3%). The patients came from Amazonas (10), Pará (01) and Rondônia (01), north States of Brazil. Five patients were infected in experimental stations of the National Institute of Amazonian Research (INPA). Although the results of this study were similar to other reports in the literature, some of the patients had more of one skin lesion. It is also reported the first case of CL caused by L. naiffi in the State of Rondônia and identified an area of disease transmission in the experimental station of INPA.

Poucos casos de leishmaniose cutânea humana (LC) causada por Leishmania naiffi foram descritos na literatura médica. Assim, o objetivo deste estudo foi relatar e analisar novos casos de infecção por L. naiffi no período compreendido entre os anos de 1992 a 2011. As cepas foram caracterizadas por análise de isoenzimas. Todos os pacientes atendidos apresentavam lesões pequenas; variando entre 1,0x1,0 mm e 13,5x11,5 mm. As lesões observadas foram amplamente distribuídas: 55,5% no membro inferior, 5,5% na área abdominal, 16,6% no membro superior e 22,2% no membro superior e para trás. Setenta e dois por cento dos pacientes tiveram lesões ulceradas. O curso clínico da doença variou de 1 a 10 meses. De acordo com o sexo, a maioria dos indivíduos infectados eram homens (83,3%). Os pacientes vieram de Amazonas (10), Pará (01) e Rondônia (01), estados do norte do Brasil. Cinco pacientes foram infectados em estações experimentais do Instituto Nacional de Pesquisas da Amazônia (INPA). Embora os resultados encontrados neste estudo fossem semelhantes às outras descrições na literatura, alguns dos pacientes apresentavam mais de uma lesão cutânea. Também é relatado o primeiro caso de LC causada por L. naiffi no Estado de Rondônia e identificada uma área de transmissão da doença na estação experimental do INPA.

Uso de dipirona como analgésico no pós-operatório de cães / The use of dipyrone as an analgesic in the post-operative of dogs

A dipirona é uma das drogas mais utilizadas no tratamento da dor de pequenos animais, especialmente de cães. Seu efeito analgésico é atribuído a seus dois principais metabólitos ativos (MAA e AA), os quais inibem as isoenzimas COX-1 e 2, além de bloquearem a síntese central e periférica de PGE2 e a ativação dos sistemas opioide e canabinoide. Seu uso em cães parece ser seguro, não havendo relatos de efeitos adversos importantes nessa espécie. Embora vários estudos acessando os efeitos analgésicos da dipirona em humanos e roedores estejam disponíveis, poucos trabalhos sobre seu uso em cães foram publicados. O objetivo deste trabalho foi realizar uma revisão de literatura sobre o uso de dipirona como analgésico no pós-operatório de cães. Os resultados da busca permitem concluir que o tratamento com dipirona promove analgesia pós-cirúrgica quando a dor é de intensidade leve a moderada, mas que, quando combinada com outras drogas analgésicas, seu efeito tende a ser mais evidenciado.(AU)

Dipyrone is one of the drugs most commonly used in the treatment of small animal pain, especially dogs. Its analgesic effect is attributed to its two main active metabolites (MAA and AA), which inhibit the COX-1 and 2 isoenzymes, besides blocking the central and peripheral synthesis of PGE2 and the activation of the opioid and cannabinoid systems. Its use in dogs appears to be safe and there are no reports of significant adverse effects in this species. Although several studies addressing the analgesic effects of dipyrone in humans and rodents are available, few papers on its use in dogs have been published. The aim of this work was to perform a literature review on the use of dipyrone as an analgesic in the postoperative period of dogs. The results of the search endorse the conclusion that dipyrone treatment promotes post-surgical analgesia when the pain is of mild to moderate intensity, but when combined with other analgesic drugs, its effect tends to be more evident.(AU)

Uso de dipirona como analgésico no pós-operatório de cães / The use of dipyrone as an analgesic in the post-operative of dogs

A dipirona é uma das drogas mais utilizadas no tratamento da dor de pequenos animais, especialmente de cães. Seu efeito analgésico é atribuído a seus dois principais metabólitos ativos (MAA e AA), os quais inibem as isoenzimas COX-1 e 2, além de bloquearem a síntese central e periférica de PGE2 e a ativação dos sistemas opioide e canabinoide. Seu uso em cães parece ser seguro, não havendo relatos de efeitos adversos importantes nessa espécie. Embora vários estudos acessando os efeitos analgésicos da dipirona em humanos e roedores estejam disponíveis, poucos trabalhos sobre seu uso em cães foram publicados. O objetivo deste trabalho foi realizar uma revisão de literatura sobre o uso de dipirona como analgésico no pós-operatório de cães. Os resultados da busca permitem concluir que o tratamento com dipirona promove analgesia pós-cirúrgica quando a dor é de intensidade leve a moderada, mas que, quando combinada com outras drogas analgésicas, seu efeito tende a ser mais evidenciado.

Dipyrone is one of the drugs most commonly used in the treatment of small animal pain, especially dogs. Its analgesic effect is attributed to its two main active metabolites (MAA and AA), which inhibit the COX-1 and 2 isoenzymes, besides blocking the central and peripheral synthesis of PGE2 and the activation of the opioid and cannabinoid systems. Its use in dogs appears to be safe and there are no reports of significant adverse effects in this species. Although several studies addressing the analgesic effects of dipyrone in humans and rodents are available, few papers on its use in dogs have been published. The aim of this work was to perform a literature review on the use of dipyrone as an analgesic in the postoperative period of dogs. The results of the search endorse the conclusion that dipyrone treatment promotes post-surgical analgesia when the pain is of mild to moderate intensity, but when combined with other analgesic drugs, its effect tends to be more evident.

Physiological performance and electrophoretic pattern of isoenzymes in Phaseolus vulgaris Lam. seeds treated with essential oil of Moringa oleifera Lam / Desempenho fisiológico e padrão eletroforético de isoenzimas em sementes de Phaseolus vulgaris Lam. tratadas com óleo essencial de Moringa oleifera Lam

In order to investigate the effect of different doses of essential oil of Moringa oleifera (EOM) on physiological potential and the expression of isoenzymes in bean seeds, an experiment was conducted with two cultivars (Colibri and Campos Gerais) and seven treatments (bactericidal, fungicide and the doses 0, 0.1, 0.2, 0.4 and 0.8% EOM). Germination, first count of germination, germination speed index (GSI), emergence and emergence rate index (ERI) were evaluated in addition to the enzymes esterase (EST), alcohol dehydrogenase (ADH), superoxide dismutase (SOD) and isocitrate lyase (ICL) by electrophoresis. In an experiment conducted in the laboratory, the dose containing 0.4% EOM promoted germination and higher averages for first germination in seeds and GSI in the Colibri cultivar. However, the highest dose used (0.8%) was responsible for the worst physiological performance by the cultivar. For Campos Gerais, increasing doses of essential oil stimulated germination, first count of germination and GSI. In the field, higher doses of essential oil gave higher germination percentages for the Colibri cultivar, not influencing the seeds of Campos Gerais. The ERI was not altered by the treatments with OEM and changes in the molecular forms of the isoenzymes were observed, with low expression values of bands for EST, ICL and ADH-related vigor, the germination of both cultivars, as well as increased expression of SOD, which is associated with reductions in germination.(AU)

A fim de investigar o efeito de diferentes doses de óleo essencial de Moringa oleifera (OEM) no potencial fisiológico e expressão de isoenzimas em sementes de feijão, conduziu-se um experimento com duas cultivares (Campos Gerais e Colibri) e sete tratamentos (bactericida, fungicida e as doses 0, 0.1, 0.2, 0.4 e 0.8% de óleo essencial). Foram avaliados a germinação, primeira contagem de germinação, índice de velocidade de germinação (IVG), emergência e índice de velocidade de emergência (IVE), além das enzimas esterase (EST), álcool desidrogenase (ADH), superóxido dismutase (SOD) e isocitrato liase (ICL) por meio de eletroforese. Em experimento conduzido em laboratório, a dose contendo 0.4% de OEM promoveu incremento na germinação e maiores médias para primeira contagem de germinação e IVG nas sementes da cultivar Colibri. No entanto, a maior dose utilizada (0.8%) foi responsável pelo pior desempenho fisiológico para a cultivar. Para Campos Gerais, as doses crescentes de óleo essencial estimularam a germinação, a primeira contagem de germinação e IVG. À campo, as doses de óleo essencial conferiram maior porcentagem de emergência para a cultivar Colibri, não influenciando as sementes da cultivar Campos Gerais. O IVE não sofreu alteração e modificações nas formas moleculares das isoenzimas foram observadas, estando os baixos valores de expressão de bandas para EST, ICL e ADH, relacionadas ao maior vigor e germinação das sementes de ambas as cultivares, bem como, maior expressão de SOD, associada à redução na germinação.(AU)

Preliminary studies of new strains of Trametes sp. from Argentina for laccase production ability

Oxidative enzymes secreted by white rot fungi can be applied in several technological processes within the paper industry, biofuel production and bioremediation. The discovery of native strains from the biodiverse Misiones (Argentina) forest can provide useful enzymes for biotechnological purposes. In this work, we evaluated the laccase and manganese peroxidase secretion abilities of four newly discovered strains of Trametes sp. that are native to Misiones. In addition, the copper response and optimal pH and temperature for laccase activity in culture supernatants were determined.The selected strains produced variable amounts of laccase and MnP; when Cu2+ was added, both enzymes were significantly increased. Zymograms showed that two isoenzymes were increased in all strains in the presence of Cu2+. Strain B showed the greatest response to Cu2+ addition, whereas strain A was more stable at the optimal temperature and pH. Strain A showed interesting potential for future biotechnological approaches due to the superior thermo-stability of its secreted enzymes.(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)