Use of PCR-DHPLC with fluorescence detection for the characterization of the bacterial diversity during cassava (Manihot esculenta Crantz) fermentation.

Denaturing high-performance liquid chromatography (DHPLC) has been described as a suitable method to study DNA polymorphisms. Here, cassava (Manihot esculenta Crantz) fermentation liquor was examined using DHPLC analysis to characterize the bacterial diversity during the fermentation process. GC-clamped amplicons corresponding to a variable region of the bacterial community 16S rDNA were synthesized using polymerase chain reaction (PCR) and then resolved on a base-composition basis using preparative DHPLC. Eluate fractions were collected at random and used as a source of whole community DNA that could be used to determine the bacterial diversity. As a first approach, GC-clamps were removed from the eluted DNA fragments using PCR to avoid the possible bias these clamps could cause during the construction of clone libraries. As a second approach, a clone library of each eluate sample was constructed, preserving the GC-clamps of the DNA fragments. The first approach generated 132 bacterial rDNA sequences with an average size of 200 bp, 45% of which had similarity to unculturable or non-classified bacteria. The second approach produced 194 sequences identified as Proteobacteria (48%), uncultured or non-classified environmental bacteria (40%) and Firmicutes (12%). We detected a remarkably greater bacterial diversity using the first approach than the second approach. The DHPLC-PCR method allowed for the fast and non-laborious detection of a vast bacterial diversity that was associated with cassava fermentation, and we conclude that it is a promising alternative for the characterization of the overall microbial diversity in complex samples.

Inseticidas neonicotinoides no controle do bicudo-do-algodoeiro Anthonomus grandis (Boheman, 1843) (Coleoptera: Curculionidae) e a falha de controle do endosulfan / Neonicotinoid insecticides for boll weevil Anthonomus grandis (Boheman, 1843) (Coleoptera: Curculionidae) control and the control failure of endosulfan

O bicudo Anthonomus grandis (Boheman 1843) (Coleoptera: Curculionidae) é considerado a principal praga do algodoeiro no Brasil e uma das principais táticas de controle é o uso de inseticidas químicos. O desenvolvimento de produtos com modos de ação diferentes dos produtos convencionais é essencial para o manejo da resistência da praga e opções de controle químico. Este trabalho teve como objetivo avaliar a eficácia de alguns inseticidas neonicotinoides no controle do inseto. Os seguintes tratamentos foram aplicados a campo em 4 pulverizações com intervalos de 5 dias (9,8% de botões atacados no início do estudo): 1) Imidacloprid 200 SC (525 mL); 2) Imidacloprid 200 SC (700 mL); 3) Imidacloprid 200 SC (1.050 mL); 4) Thiamethoxam 250 WG (300 g); 5) Acetamiprid 200 SP (300 g); 6) Clothianidin 600 SC (262,5 mL); 7) Thiacloprid 480 SC (300 mL); 8) Endosulfan 350 CE (2.000 mL); e 9) Testemunha não tratada. O delineamento experimental foi o de blocos ao acaso com nove tratamentos e quatro repetições. As médias dos botões florais atacados por tratamento foram submetidas à análise de variância e comparadas pelo teste de Tukey (? = 0,05), enquanto que para o cálculo da eficiência dos inseticidas foi utilizada a fórmula de Abbott. Clothianidin 600 SC (262,5 mL p.c./ha) foi o neonicotinoide que apresentou a maior eficiência de controle (73% de controle aos seis dias após a terceira aplicação do inseticida). O ciclodieno Endosulfan 350 CE (2.000 mL do p.c./ha), utilizado como produto padrão do produtor, não controlou satisfatoriamente a praga como frequentemente citado. Não houve diferenças estatisticamente significativas entre os tratamentos para os parâmetros de produtividade (peso de algodão em caroço e número de capulhos por planta).

The boll weevil Anthonomus grandis (Boheman 1843) (Coleoptera: Curculionidae) is the most important cotton pest in Brazil, and insecticides play an important role in controlling its adults, so the development of products with new modes of action is essential to achieve successful control alternatives and resistance management options. The objective of this study was to evaluate the efficacy of some neonicotinoid insecticides to control the pest. The experimental design consisted of completely randomized blocks, with nine treatments and four replications. The following treatments (dosage of the commercial product per hectare) were applied in the field (with 9.8% of damaged squares at the beginning of the study): 1) Imidacloprid 200 SC (525 mL); 2) Imidacloprid 200 SC (700 mL); 3) Imidacloprid 200 SC (1,050 mL); 4) Thiamethoxam 250 WG (300 g); 5) Acetamiprid 200 SP (300 g); 6) Clothianidin 600 SC (262,5 mL); 7) Thiacloprid 480 SC (300 mL); 8) Endosulfan 350 CE (2,000 mL); and 9) Untreated control. The efficacy of insecticides was calculated with the Abbott formula; the data of damaged squares in each treatment was submitted to variance analysis and its means compared by Tukey test (? = 0.05). The results showed that Clothianidin 600 SC (262.5 mL/ha) was the most efficient neonicotinoid and reached 73% of efficacy at 6 days after the third application. The cyclodiene Endosulfan 350 CE (2,000 mL/ha), which was the standard farmer treatment, did not control the boll weevil as frequently cited. There were no significant differences in cotton production (fiber plus seed and bolls per plant) among treatments.

Inseticidas neonicotinoides no controle do bicudo-do-algodoeiro Anthonomus grandis (Boheman, 1843) (Coleoptera: Curculionidae) e a falha de controle do endosulfan

ABSTRACT The boll weevil Anthonomus grandis (Boheman 1843) (Coleoptera: Curculionidae) is the most important cotton pest in Brazil, and insecticides play an important role in controlling its adults, so the development of products with new modes of action is essential to achieve successful control alternatives and resistance management options. The objective of this study was to evaluate the efficacy of some neonicotinoid insecticides to control the pest. The experimental design consisted of completely randomized blocks, with nine treatments and four replications. The following treatments (dosage of the commercial product per hectare) were applied in the field (with 9.8% of damaged squares at the beginning of the study): 1) Imidacloprid 200 SC (525 mL); 2) Imidacloprid 200 SC (700 mL); 3) Imidacloprid 200 SC (1,050 mL); 4) Thiamethoxam 250 WG (300 g); 5) Acetamiprid 200 SP (300 g); 6) Clothianidin 600 SC (262,5 mL); 7) Thiacloprid 480 SC (300 mL); 8) Endosulfan 350 CE (2,000 mL); and 9) Untreated control. The efficacy of insecticides was calculated with the Abbott formula; the data of damaged squares in each treatment was submitted to variance analysis and its means compared by Tukey test (? = 0.05). The results showed that Clothianidin 600 SC (262.5 mL/ha) was the most efficient neonicotinoid and reached 73% of efficacy at 6 days after the third application. The cyclodiene Endosulfan 350 CE (2,000 mL/ha), which was the standard farmer treatment, did not control the boll weevil as frequently cited. There were no significant differences in cotton production (fiber plus seed and bolls per plant) among treatments.

RESUMO O bicudo Anthonomus grandis (Boheman 1843) (Coleoptera: Curculionidae) é considerado a principal praga do algodoeiro no Brasil e uma das principais táticas de controle é o uso de inseticidas químicos. O desenvolvimento de produtos com modos de ação diferentes dos produtos convencionais é essencial para o manejo da resistência da praga e opções de controle químico. Este trabalho teve como objetivo avaliar a eficácia de alguns inseticidas neonicotinoides no controle do inseto. Os seguintes tratamentos foram aplicados a campo em 4 pulverizações com intervalos de 5 dias (9,8% de botões atacados no início do estudo): 1) Imidacloprid 200 SC (525 mL); 2) Imidacloprid 200 SC (700 mL); 3) Imidacloprid 200 SC (1.050 mL); 4) Thiamethoxam 250 WG (300 g); 5) Acetamiprid 200 SP (300 g); 6) Clothianidin 600 SC (262,5 mL); 7) Thiacloprid 480 SC (300 mL); 8) Endosulfan 350 CE (2.000 mL); e 9) Testemunha não tratada. O delineamento experimental foi o de blocos ao acaso com nove tratamentos e quatro repetições. As médias dos botões florais atacados por tratamento foram submetidas à análise de variância e comparadas pelo teste de Tukey (? = 0,05), enquanto que para o cálculo da eficiência dos inseticidas foi utilizada a fórmula de Abbott. Clothianidin 600 SC (262,5 mL p.c./ha) foi o neonicotinoide que apresentou a maior eficiência de controle (73% de controle aos seis dias após a terceira aplicação do inseticida). O ciclodieno Endosulfan 350 CE (2.000 mL do p.c./ha), utilizado como produto padrão do produtor, não controlou satisfatoriamente a praga como frequentemente citado. Não houve diferenças estatisticamente significativas entre os tratamentos para os parâmetros de produtividade (peso de algodão em caroço e número de capulhos por planta).