Sobrevivência e reprodução de Synthesiomyia nudiseta (Diptera, Muscidae)

A reprodução e a sobrevivência de Synthesiomyia nudiseta (Wulp, 1883) foram estudadas entre 22 e 28ºC, UR de 70% e fotoperíodo de 12h. Os dados foram analisados através de classes de idade, tabelas de vida de fecundidade, análise de sobrevivência pela distribuição de Weibull e análise das curvas de sobrevivência através do conceito de entropia (H). A taxa intrínseca de aumento natural (r m), taxa finita de aumento (lambda), taxa de reprodução líquida (R0) e tempo médio de uma geração (T) foram 0,534; 1,7; 59,439 e 7,65 respectivamente, sugerindo rápido crescimento populacional nestas condições. Isso foi causado pela alta fecundidade, confirmando o que ocorre para esta espécie, com média de 266 ovos/fêmea e período de pré-oviposição de 10,25 dias. O modelo de Weibull demonstrou que a sobrevivência não foi constante ao longo da vida de adultos, com mortalidade pronunciada depois da quarta e quinta semanas e valores de H intermediários aos valores teóricos esperados (0,0-0,5) para os ambos sexos, demonstrando que a curva de sobrevivência é retangular (com maior impacto da mortalidade na quarta e quinta semanas após a emergência). Os resultados apresentados podem ser considerados como base para estudos pormenorizados da dinâmica e do crescimento populacional desta espécie em hábitats naturais.

Characterization of a Trypanosoma cruzi genomic fragment complementary to several species-specific mRNAs but different from the spliced leader sequence

We have isolated a clone of Trypanosoma cruzi genimic DNA, lambda 3b2-5, which contains sequences that are reiterated in the genome. Northtern blot analysis showed that clone 3b2-5 hybridizes to 1,200-5,000 bases different mRNA species. The number of mRNAs species hybridized to clone 3b2-5 exceeds its coding capacity showing that this clone carries sequences that are common to several mRNAs species and conserved in the poly A(+) RNA. These sequences are not homologous to the T. cruzi spliced leader sequence, since clone 3b2-5 hybridize to a synthetic 20 nucleotice complementary to the spliced leader sequence. Clone 3b2-5 does not hybridize to DNA and RNA from several genera of Trypanosomatidae and other Trypanosoma species indicating that it carries T. cruzi species-specific sequences.

The p53 gene expression and its developmental regulation in schistosomes

We have studied the gene expression, especially of the oncoproteins, and its regulation in schistosomes. Schistosomes have a complex life cycle with defined dimorphic lifestyle. The parasite are so far unique in biology in expressing oncogene products in their adult stage. In order to characterize the expression and developmental regulation, a lambda gt 11 cDNA library and lambda EMBL4 genomic DNA library of each growth stage of Schistosoma mansoni and S. japonicum was constructed, and was screened with various monoclonal antibodies against ongogene products. One positive plaque reacted to anti-p53 antibody (Ab-2, Oncogene Science, Inc.) was further analyzed. This fusion protein was about 120 KDa in molecular weights, and expressed as 1.4 Kb RNA in the adult stage. P53 gene is well-known as the negative regulator of the cell cicle, and the mutations in the gene are turning out to be the most common genetic alterations in human cancers. The comparison of the gene structure among species and stages were being conducted. Chromosome structures, C-band formation, and the results of in situ hybridization using the phage probe would be discussed.

Cloning, Expression and Toxicity of a Mosquitocidal Toxin Gene of Bacillus thuringiensis subsp. medellin

Bacillus thuringiensis (Bt) subsp. medellin (Btmed) produces parasporal crystalline inclusions which are toxic to mosquito larvae. It has been shown that the inclusions of this bacterium contain mainly proteins of 94, 68 and 28-30 kDa. EcoRI partially digested total DNA of Btmed was cloned by using the Lambda Zap II cloning kit. Recombinant plaques were screened with a mouse policlonal antibody raised against the 94 kDa crystal protein of Btmed. One of the positive plaques was selected, and by in vivo excision, a recombinant pBluescript SK(-) was obtained. The gene encoding the 94 kDa toxin of Btmed DNA was cloned in a 4.4 kb DNA fragment. Btmed DNA was then subcloned as a EcoRI/EcoRI fragment into the shuttle vector pBU4 producing the recombinant plasmid pBTM3 and used to transform by electroporation Bt subsp. israelensis (Bti) crystal negative strain 4Q2-81. Toxicity to mosquito larvae was estimated by using first instar laboratory reared Aedes aegypti, and Culex quinquefasciatus larvae challenged with whole crystals. Toxicity results indicate that the purified inclusions from the recombinant Bti strain were toxic to all mosquito species tested, although the toxicity was not as high as the one produced by the crystal of the Btmed wild type strain. Poliacrylamide gel electrophoresis indicate that the inclusions produced by the recombinant strain Bti (pBTM3) were mainly composed of the 94 kDa protein of Btmed, as it was determined by Western blot

Life cycle and reproductive parameters of Clerada apicicornis signoret (Hemiptera: Lygaeidae) under laboratory conditions

The life cycle of Clerada apicicornis was determined under laboratory conditions. Mean development times in days were: egg 27.2, nymph I 12.5, nymph II 12, nymph III 13.4, nymph IV 16.4, nymph V 26. The life expectancy of adults ranged from 117 to 317 days (mean 196 days). Based on a cohort of 29 females of C. apicicornis, a horizontal life table was constructed. The following predictive parameters were obtained: net rate of reproduction (Ro = 48.31), intrinsic rate of population increase (r m = 0.153), generation time (Tc = 28.20 weeks), and finite rate of population increment (lambda = 1.16). The reproductive value (Vx) for each age class of the cohort females was calculated. The following observed parameters were calculated after mortality in each stage: net rate of reproduction (R'o=13.4), intrinsic rate of population increase (r c' =0.09 ), and finite rate of population increment (lambda' =1.1). The generation time (Tc' =27.4) was estimated using the methods of Laughlin and Bengstron. A vertical life table was elaborated and mortality was described for one generation of the cohort.

Comparative evaluation of pyrethroid insecticide formulations against Triatoma infestans (Klug): residual efficacy on four substrates

We investigated the residual efficacy of four insecticide formulations used in Chagas disease vector control campaigns: cyfluthrin 12.5% suspension concentrace (SC), lambda-cyhalothrin 10% wettable powder (WP), deltamethrin 2.5% SC, and 2.5% WP on four types of circular blocks of wood, straw with mud, straw with mud painted with lime, and mud containing 5% of cement. Three concentrations of these insecticides were tested: the LC90 (previously determined on filter paper), the double of the LC90, and the recommended operational dose. For each bioassay test, 15 third-stage nymphs of Triatoma infestans (Klug) (Hemiptera: Reduviidae) were exposed for 120 h to each treatment at 24 h, 30, 60, 90, and 180 days post-spraying. Mortality rates, moulting history and behaviour were recorded at 24, 48, 72, and 120 h of exposure. Mortality rates were highest during the first 30 days post-spraying. Highest mortality rates (above 50%) were observed for deltamethrin 2.5% SC and lambda-cyhalothrin 10% WP on wood blocks up to three months post-spraying. Mud was the substrate on which treatments showed lowest persistence, with the other two substrates showing intermediate residual efficacy of all treatments. During the first 30 days WP formulations were not as effective as SC flowable formulations but, overall in the longer term, WP gave grater mortality rates of T. infestans nymphs exposed at up to six months post-spraying. Porous surfaces, especially mud, showed most variability presumably due to absorption of the insecticide. In contrast the less porous surfaces (i.e. wood and lime-coated mud) kept mortality rates high for longer post-treatment, irrespective of the insecticide concentration used.

Toxicity of pyrethroids and repellency of diethyltoluamide in two deltamethrin-resistant colonies of Triatoma infestans Klug, 1834 (Hemiptera: Reduviidae)

The aim of the currrent investigation was to evaluate (a) the toxicity of three pyrethroids (deltamethrin, lambda-cyhalothrin, and tetramethrin); (b) the effect of these insecticides on the locomotor activity; and (c) the repellent effect of N,N-diethyl-m-toluamide (DEET) on two deltamethrin-resistant strains of Triatoma infestans from Argentina (El Chorro and La Toma), and one susceptible strain. The resistance ratios (RRs) obtained for the La Toma strain were: > 10,769, 50.7, and > 5.2 for deltamethrin, lambda-cyhalothrin, and tetramethrin respectively. The RRs for the El Chorro strain were: > 10,769, 85.8, and > 5.2 for deltamethrin, lambda-cyhalothrin, and tetramethrin respectively. The hyperactivity usually caused by the three pyrethroids was in both the deltamethrin-resistant strains compared to the susceptible reference strain. No differences were observed in the repellent effect of DEET between the three groups. These results indicate that the deltamethrin-resistant insects have a cross resistance to lambda-cyhalothrin and tetramethrin, and are also resistant to the first symptom of pyrethroid poisoning (hyperactivity). However, the sensorial process related to DEET repellency does not appear to be altered.

Association of cytokine genetic polymorphism with hepatites B infection evolution in adult patients

The infection by the hepatitis B virus (HBV) has different forms of evolution, ranging from self-limited infection to chronic hepatic disease. The objective of this study was to evaluate the influence of cytokine genetic polymorphisms in the disease evolution. The patients were divided into two groups, one with chronic HBV (n = 30), and the other with self-limited infection (n = 41). The genotyping for TNF (-308), TGFB1 (+869, +915), IL-10 (1082, -819, and -592), IL-6 (-174), and IFNG (+874) was accomplished by the PCR-SSP (polymerase chain reaction with sequence specific primers technique using the One Lambda kit. Although no statistically significant differences were found between the groups, the combination of TNF -308GG and IFNG +874TA was found in a lower frequency in chronic patients than in individuals with self-limited infection (26.7 versus 46.3%; P = 0.079; OR = 0.40; IC95% = 0.14-1.11). In chronic patients with histological alterations it was not observed the genotype TGFB1+869 C/C, against 24.4% in the self limited infection group (100 versus 75.6%; P = 0.096; OR = 7.67; IC95% = 0.42-141.63). Further studies in other populations, and evaluation of a greater number of individuals could contribute for a better understanding of the cytokine genetic polymorphism influence in HBV infection evolution.

Capillaria spp. eggs in Patagonian archaeological sites: statistical analysis of morphometric data

Discriminant analysis was used to identify eggs of Capillaria spp. at specific level found in organic remains from an archaeological site in Patagonia, Argentina, dated of 6,540 ± 110 years before present. In order to distinguish eggshell morphology 149 eggs were measured and grouped into four arbitrary subsets. The analysis used on egg width and length discriminated them into different morphotypes (Wilks' lambda = 0.381, p < 0.05). The correlation analysis suggests that width was the most important variable to discriminate among the Capillaria spp. egg morphotypes (Pearson coefficient = 0.950, p < 0.05). The study of eggshell patterns, the relative frequency in the sample, and the morphometric data allowed us to correlate the four morphotypes with Capillaria species.

Tabela de vida de fertilidade de Lysiphlebus testaceipes (Cresson) (Hymenoptera, Braconidae, Aphidiinae) em Rhopalosiphum maidis (Fitch) e Aphis gossypii Glover (Hemiptera, Aphididae)

Avaliar o potencial de crescimento de Lysiphlebus testaceipes (Cresson) é importante para seu uso em programas de controle biológico de pulgões. Este trabalho teve como objetivo determinar a tabela de vida de fertilidade de L. testaceipes em Rhopalosiphum maidis (Fitch) e Aphis gossypii Glover. Na avaliação da mortalidade de imaturos, do desenvolvimento e da razão sexual foram utilizadas 12 fêmeas do parasitóide e 480 ninfas de cada pulgão testado. Na avaliação da longevidade e da fertilidade foram utilizadas 15 fêmeas do parasitóide e uma colônia por dia de cada pulgão, até a morte da fêmea do parasitóide, sendo 300 (1º dia); 250 (2º dia); 200 (3º dia); 150 (4º dia); 100 (5º dia) e 50 ninfas nos demais dias. L. testaceipes apresentou taxas de mortalidade de imaturos de 5,6% em R. maidis e de 9,2% em A. gossypii, desenvolvimento de 10,2 e 10,1 dias e razão sexual de 0,71 e 0,66, respectivamente. L. testaceipes apresentou fecundidade de 498,2 ovos em R. maidis e de 327,8 ovos em A. gossypii. Os parâmetros de crescimento de L. testaceipes em R. maidis e A. gossypii foram, respectivamente, R O= 205,38 e 164,08 fêmeas; r m= 0,449 e 0,431 fêmeas/fêmeas/dia; lambda = 1,57 e 1,54 fêmeas/dia; T= 11,86 e 11,83 dias e TD= 10,78 e 11,27 dias. L. testaceipes apresenta alto potencial de crescimento em R. maidis e A. gossypii. R. maidis mostrou ser hospedeiro adequado aos propósitos de criação massal e utilização em sistema de criação aberta para L. testaceipes.

Resíduos de agrotóxicos em frutos de tomate

O uso intensivo de agrotóxicos em culturas de tomates tem causado preocupações quanto à provável contaminação do produto final. Este trabalho teve como objetivo monitorar, por dois anos consecutivos, em quatro propriedades de uma área altamente tecnificada, o uso de agrotóxicos em tomate irrigado durante seu desenvolvimento, quantificar os resíduos dos principais princípios ativos utilizados, e avaliar o grau de contaminação dos frutos colhidos. Foram analisados metamidofós, clorpirifós, captan, clorotalonil, endosulfan, lambda-cialotrina, e cobre. Para avaliar o grau de contaminação dos frutos foi validado um método de análise de resíduos múltiplos capaz de quantificar esses compostos. Os resíduos encontrados foram de fungicidas e inseticidas aplicados nas fases de frutificação e maturação: captan, 0,35 mg/kg, na Fazenda 2; clorotalonil, 0,16 mg/kg e 0,95 mg/kg, nas Fazendas 1 e 2, respectivamente; lambda-cialotrina, 0,03 mg/kg, na Fazenda 2; cobre, 2,03 mg/kg, 3,75 mg/kg e 1,44mg/kg, nas Fazendas 1, 2 e 3, respectivamente, e 0,95 mg/kg, 1,70 mg/kg e 2,31 mg/kg, na Fazenda 4. Não ocorreram resíduos dos inseticidas organofosforados, aplicados principalmente durante o desenvolvimento vegetativo da cultura. Não houve contaminação dos tomates pelos agrotóxicos analisados. Os resíduos que devem ser monitorados são os dos agrotóxicos aplicados na fase de maturação da cultura.

Monitoramento e avaliação do risco de contaminação por pesticidas em água superficial e subterrânea na região de Guaíra

O objetivo deste trabalho foi determinar os níveis de ocorrência de pesticidas em água, tendo em vista preservar a água de contaminação por esses produtos na região de Guaíra, SP. A hipótese foi que o uso intensivo de pesticidas nessa região propiciaria a contaminação das águas superficiais e subterrâneas por seus resíduos. Para a caracterização das propriedades, das características e da dinâmica da água do solo, foi selecionada a Fazenda Macaúba por possuir relevo e solos característicos da região. Aproximadamente 80% da área da fazenda é constituída por Latossolo Vermelho distroférrico típico (LVdf), profundo. As áreas cultivadas apresentam um horizonte superficial (Ap) completamente desprovido de sua estrutura natural e, abaixo deste, um horizonte compactado, ambos frutos das práticas agrícolas. Durante dois anos e meio realizou-se o monitoramento do Ribeirão do Jardim que abastece a cidade de Guaíra, bem como de um de seus afluentes e da água subterrânea proveniente do Aqüífero Guarani, à procura de pesticidas. A água superficial foi coletada de 21em 21 dias e a subterrânea, no início e no final do período monitorado (dois anos e meio). Os produtos monitorados, trifluralina, endosulfan, lambda cialotrina, dicofol (4,4 diclorobenzofenona), captan, metil paration, clorotalonil e clorpirifós, foram selecionados a partir de um levantamento dos pesticidas utilizados em quatro áreas agrícolas localizadas na bacia do Ribeirão do Jardim e em razão da efetividade de um método de análise de resíduos múltiplos para sua quantificação. Os resultados analíticos indicaram que não houve contaminação da água subterrânea, mas que ocasionalmente houve contaminação direta das águas de superfície. A não-contaminação da água subterrânea deve-se principalmente às características dos Latossolos, como sua grande espessura, sua textura argilosa e sua grande capacidade de armazenamento de água.

Seleção de atrativos alimentares e toxicidade de inseticidas para o manejo da broca-pequena-do-tomateiro

O objetivo deste trabalho foi avaliar a preferência alimentar, o limiar de ingestão e efeito tóxico inseticidas associados atrativos, em adultos Neoleucinodes elegantalis. Foram testados os atrativos: melado e mel a 10%, extrato hexânico de frutos verdes de tomate a 0,4%, sacarose a 5%, suco de laranja e suco de uva a 30%, vinagre de vinho tinto a 10% e proteína hidrolisada a 5%. Com base no teste de preferência alimentar, foram selecionados os atrativos sacarose, melado, mel e suco de laranja, para determinar o limiar de concentração capaz de estimular a alimentação de adultos de N. elegantalis. Foi testado o efeito tóxico de inseticidas associados ao mel a 10%. A sacarose e o mel apresentaram o melhor resultado em relação ao número de pousos e ao tempo de pouso e de alimentação de adultos de N. elegantalis. Os inseticidas não afetaram negativamente a atração pelo alimento dos adultos de N. elegantalis. Carbaril, cartape, deltametrina, fenpropatrina, indoxacarbe, lambda-cialotrina e lufenurom provocaram 100% de mortalidade em adultos (machos + fêmeas), após 24 horas de exposição, e mostraram-se promissores para o uso em iscas tóxicas.

Characterization and multivariate classification of grapes and wines of two Cabernet Sauvignon clones

The objective of this work was to assess and characterize two clones, 169 and 685, of Cabernet Sauvignon grapes and to evaluate the wine produced from these grapes. The experiment was carried out in São Joaquim, SC, Brazil, during the 2009 harvest season. During grape ripening, the evolution of physical-chemical properties, phenolic compounds, organic acids, and anthocyanins was evaluated. During grape harvest, yield components were determined for each clone. Individual and total phenolics, individual and total anthocyanins, and antioxidant activity were evaluated for wine. The clones were also assessed regarding the duration of their phenological cycle. During ripening, the evolution of phenolic compounds and of physical-chemical parameters was similar for both clones; however, during harvest, significant differences were observed regarding yield, number of bunches per plant and berries per bunch, leaf area, and organic acid, polyphenol, and anthocyanin content. The wines produced from these clones showed significant differences regarding chemical composition. The clones showed similar phenological cycle and responses to bioclimatic parameters. Principal component analysis shows that clone 685 is strongly correlated with color characteristics, mainly monomeric anthocyanins, while clone 169 is correlated with individual phenolic compounds.

Protective effect and economic impact of insecticide application methods on barley

The objective of this work was to evaluate the protective effect of different forms of insecticide application on the transmission of yellow dwarf disease in barley cultivars, as well as to determine the production costs and the net profit of these managements. The experiments were carried out during 2011 and 2012 growing seasons, using the following managements at main plots: T1, seed treatment with insecticide (ST) + insecticide on shoots at 15-day interval; T2, just ST; T3, insecticide applied on shoots, when aphid control level (CL) was reached; T4, without insecticide; and T5, ST + insecticide on shoots when CL was reached. Different barley cultivars - BRS Cauê, BRS Brau and MN 6021 - were arranged in the subplots. Insecticides lambda cyhalothrin (pyrethroid) and thiamethoxam (neonicotinoid) were used. There were differences on yellow dwarf disease index in both seasons for the different treatments, while damage to grain yield was influenced by year and aphid population. Production costs and net profit were different among treatments. Seed treatment with insecticide is sufficient to reduce the transmission of yellow dwarf disease in years with low aphid population pressure, while in years with larger populations, the application of insecticide on shoots is also required.