Determination of the median lethal concentration (LC(50)) of mycoinsecticides for the control of Ceratitis capitata (Diptera: Tephritidae)

The aim of this study was to determine the median lethal concentration (LC(50)) of the commercial products Boveril WP (R) (Beauveria bassiana) and Metarril WP (R) (Metarhizium anisopliae) on the larvae and pupae of the fruit Ceratitis capitata. Insects used in this study came from a laboratory colony. The evaluated product concentrations were 10.00, 15.00, 20.00 and 25.00 g/L of water, which correspond, respectively, to 5.00x10(9), 7.50x10(9), 10.00x10(9) and 12.50x10(9) viable conidia/L of water for the two products, and in the control only water was applied. Third instar larvae and pupae of C. capitata were used in this study. Results showed an overall mortality of larvae with all conidial concentrations of M. anisopliae. The LC(50) values for larvae were 2.99 and 2.97 g/L for Boveril (R) and Metarril (R), respectively, while for pupae they were 3.12 and 4.74 g/L for Boveril (R) and Metarril (R), respectively. The high pathogenicity demonstrated by lower conidial concentrations of the tested products may mean greater efficiency from both economic and environmental points of view.

Chronic exposure to sub-lethal concentration of a glyphosate-based herbicide alters hormone profiles and affects reproduction of female Jundia (Rhamdia quelen)

This work was carried out to verify the effect of a glyphosate-based herbicide on Jundia hormones (cortisol, 17 beta-estradiol and testosterone), oocyte and swim-up fry production. Earthen ponds containing Jundia females were contaminated with glyphosate (3.6 mg/L); blood samples were collected from eight females from each treatment immediately before, or at 1, 10, 20 30 and 40 days following contamination. A typical post-stress rise in cortisol levels was observed at the 20th and 40th days following exposure to glyphosate. At the 40th day, 17 beta-estradiol was decreased in the exposed females. A similar number of oocytes were stripped out from females from both groups, however, a lower number of viable swim-up fry were obtained from the herbicide exposed females, which also had a higher liver-somatic index (LSI). The results indicate that the presence of glyphosate in water was deleterious to Rhamdia quelen reproduction, altering steroid profiles and egg viability. (c) 2006 Elsevier B.V. All rights reserved.

Toxicity effect of the acaricide fipronil in semi-engorged females of the tick Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae): Preliminary determination of the minimum lethal concentration and LC50

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Median lethal concentration of formaldehyde and its genotoxic potential in bullfrog tadpoles (Lithobates catesbeianus)

In order to avoid that contaminated frog farms animals escaping in the environment and become potential vector of emergent diseases, studies with disinfection protocol are strictly necessary. The formaldehyde is one of the compounds tested in fungal disinfection protocols and also used in aquaculture. This study aimed to determine the median lethal concentration (LC50-96h) of formaldehyde in bullfrog tadpoles and to evaluate the possible genotoxic effects in acute exposition. Accordingly, the animals were exposed to formaldehyde in the concentrations of 6, 9, 12, 15, and 18 mg L(-1), and after 96 h blood samples were drawn for the micronucleus (MN) test. The LC50-96h was 10.53 mg L(-1), and the MN frequency increased in proportion to the formaldehyde concentrations, with an estimated frequency in the negative control being 1.35 MN/individual. We concluded that formaldehyde is genotoxic to tadpoles of bullfrogs in the tested concentrations, and the choice of this chemical should be contemplated before its use in animals in captivity.

Chronic exposure to sub-lethal concentration of a glyphosate-based herbicide alters hormone profiles and affects reproduction of female Jundia (Rhamdia quelen)

This work was carried out to verify the effect of a glyphosate-based herbicide on Jundia hormones (cortisol, 17 beta-estradiol and testosterone), oocyte and swim-up fry production. Earthen ponds containing Jundia females were contaminated with glyphosate (3.6 mg/L); blood samples were collected from eight females from each treatment immediately before, or at 1, 10, 20 30 and 40 days following contamination. A typical post-stress rise in cortisol levels was observed at the 20th and 40th days following exposure to glyphosate. At the 40th day, 17 beta-estradiol was decreased in the exposed females. A similar number of oocytes were stripped out from females from both groups, however, a lower number of viable swim-up fry were obtained from the herbicide exposed females, which also had a higher liver-somatic index (LSI). The results indicate that the presence of glyphosate in water was deleterious to Rhamdia quelen reproduction, altering steroid profiles and egg viability. (c) 2006 Elsevier B.V. All rights reserved.

Statistical evaluation of the fixed concentration procedure for acute inhalation toxicity assessment

The conventional method for the assessment of acute inhalation toxicity (OECD Test Guideline 403, 1981) uses death of animals as an endpoint to identify the median lethal concentration (LC50). A new OECD Testing Guideline called the Fixed Concentration Procedure (FCP) is being prepared to provide an alternative to Test Guideline 403. Unlike Test Guideline 403, the FCP does not provide a point estimate of the LC50, but aims to identify an airborne exposure level that causes clear signs of nonlethal toxicity. This is then used to assign classification according to the new Globally Harmonized System of Classification and Labelling scheme (GHS). The FCP has been validated using statistical simulation rather than byin vivo testing. The statistical simulation approach predicts the GHS classification outcome and the numbers of deaths and animals used in the test for imaginary substances with a range of LC50 values and dose response curve slopes. This paper describes the FCP and reports the results from the statistical simulation study assessing its properties. It is shown that the procedure will be completed with considerably less death and suffering than Test Guideline 403, and will classify substances either in the same or a more stringent GHS class than that assigned on the basis of the LC50 value.

Molluscicidal and ovicidal activities of plant extracts of the Piperaceae on Biomphalaria glabrata (Say, 1818)

Schistosomiasis is a tropical disease caused by Schistosoma and occurs in 54 countries, mainly in South America, the Caribbean region, Africa and the eastern Mediterranean. Currently, 5 to 6 million Brazilian people are infected and 30,000 are under infection risk. Typical of poor regions, this disease is associated with the lack of basic sanitation and very frequently to the use of contaminated water in agriculture, housework and leisure. One of the most efficient methods of controlling the disease is application of molluscicides to eliminate or to reduce the population of the intermediate host snail Biomphalaria glabrata. Studies on molluscicidal activity of plant extracts have been stimulated by issues such as environmental preservation, high cost and recurrent resistance of snails to synthetic molluscicides. The aim of this study was to determine the molluscicide action of extracts from Piperaceae species on adult and embryonic stages of B. glabrata. Fifteen extracts from 13 Piperaceae species were obtained from stems, leaves and roots. Toxicity of extracts was evaluated against snails at two different concentrations (500 and 100 ppm) and those causing 100% mortality at 100 ppm concentration were selected to obtain the LC(90) (lethal concentration of 90% mortality). Piper aduncum, P. crassinervium, P. cuyabanum, P. diospyrifolium and P. hostmannianum gave 100% mortality of adult snails at concentrations ranging from 10 to 60 ppm. These extracts were also assayed on embryonic stages of B. glabrata and those from P. cuyabanum and P. hostmannianum showed 100% ovicidal action at 20 ppm.

Antimicrobial and Cytotoxic Activity of Fruit Extract from Syzygium cumini (L.) Skeels

The aim of the present study was to evaluate the antimicrobial and cytotoxic activity of the ethanolic extract of S. cumini according to the Clinical and Laboratory Standards Institute reference method (with modifications), determining the minimal inhibitory and lethal concentration. Activity against Gram-positive (Staphylococcus aureus and S. epidermidis), Gram-negative (Pseudomonas aeruginosa) and yeast of Candida sp and Cryptococcus neoformans was evaluated. The effects of the fruit extract were examined in hamster cells ovaries in concentrations ranging from 1250.0 a 4.9 mu g/ml, measuring the reduction of the tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulphophenyl)-2H-tetrazolium. The extract showed both bactericidal and fungicidal activity among the various microorganisms tested and the MIC ranging from 7.8 to 250 mu g/ml. The MIC, MBC and MFC should values that were similar for all the microorganisms. Cytotoxicity index of the dried extract corresponded to the concentration of 400 mu g/ml. The extract could potentially be used in topical antimicrobial products. Thus, the activity of extract was potent to bacteria and mainly to non-albicans species and C. neoformans.

Extratos vegetais e Bacillus thuringiensis visando o manejo de Duponchelia fovealis Zeller (Lepidoptera: Crambidae)

A lagarta Duponchelia fovealis Zeller (Lepidoptera: Crambidae) foi relatada em plantios comerciais do morangueiro no estado do Espírito Santo ocasionando grandes problemas à cultura. Porém, por ser uma praga recente, não há registro de produtos para o seu controle. A cada dia aumenta a demanda por alimentos e outros produtos livres de resíduos, além da necessidade de uma agricultura mais desenvolvida e sustentável. Pesquisas com agentes de controle biológico e extratos vegetais surgem como alternativa para o manejo desse inseto-praga. Este trabalho teve como objetivo avaliar a eficiência de formulados comerciais à base de Bacillus thuringiensis e a atividade do uso dos extratos aquosos de alho e fumo, visando sua adoção como métodos alternativos de controle de D. fovealis. Nos bioensaios para avaliar a patogenicidade e virulência de duas formulações comerciais à base de B. thuringiensis, Agree® e Dipel WP®, sobre a dieta artificial adaptada à base de farelo de soja, germe de trigo e açúcar, proposta por King e Hartley (1985) para Diatraea saccharalis (Lepidoptera: Crambidae), foram inoculados 70 μL de cada formulado comercial, na concentração 1 x 108 esporos·mL-1. Em seguida, avaliou-se a virulência dos respectivos formulados, isso através da estimativa da concentração letal (CL50) para o estádio de maior suscetibilidade. Em virtude dos resultados encontrados, observou-se que o estágio 1 de desenvolvimento apresentou 95,88% e 86,76% de mortalidades para os produtos Agree® e Dipel WP®, respectivamente, demonstrando patogenicidade e virulência à D. fovealis. No bioensaio para avaliar a atividade dos extratos aquosos de alho e fumo, estes foram aplicados na concentração 10% (m/v). Todos os tratamentos foram pulverizados com Torre de Potter, calibrada a pressão de 15 lb/pol². Posteriormente estimou-se a concentração letal (CL50) do extrato aquoso de fumo, o qual apresentou 95% de mortalidade no teste de suscetibilidade. Desta forma, com os resultados obtidos na presente pesquisa, concluiu-se que a utilização de formulados comerciais à base de B. thuringiensis e extrato de fumo podem ser uma alternativa no manejo fitossanitário de D. fovealis.

Analysis of toxicity on Bacillus sphaericus from amazonian soils to Anopheles darlingi and Culex quinquefasciatus larvae

Bioassays under laboratory conditions aiming to determine the larvicidal activity of Bacillus sphaericus were carried out on Anopheles darlingi and Culex quinquefasciatus. In order to estimate the toxicity through median lethal concentration (LC50) and the relative potency of the strains to B. sphaericus standard strain 2362, probit analysis was performed utilizing the POLO-PC program. The findings of LC50 pointed out high effectiveness on strains IB15 (0.040 ppm), IB19 and S1116 (0.048 ppm), IB16 (0.052 ppm) and S265 (0.057 ppm). Strain IB15 presented nearly 50% more potency than strain 2362 in bioassays conducted on A. darlingi. It was observed that IB16 and S1116 strains were the most powerful against C. quinquefasciatus, showing to be about 300-400% stronger than 2362 strain. The results show that laboratory conditioned evaluation can be an important way to select promising bacteria with entomopathogenic action on biolarvicides production for use on mosquitoes breeding sites.

Tolerance to temperature, pH, ammonia and nitrite in cardinal tetra, Paracheirodon axelrodi, an amazonian ornamental fish

Poor water quality condition has been pointed out as one of the major causes for the high mortality of ornamental fishes exported from the state of Amazonas, Brazil. The purpose of the current study was to define water quality standards for cardinal tetra (Paracheirodon axelrodi), by establishing the lower and higher for lethal temperature (LT50), lethal concentration (LC50) for total ammonia and nitrite and LC50 for acid and alkaline pH. According to the findings, cardinal tetra is rather tolerant to high temperature (33.3 ºC), to a wide pH range (acid pH=2.9 and alkaline pH=8.8) and to high total ammonia concentration (23.7 mg/L). However, temperatures below 19.6 ºC and nitrite concentrations above 1.1 mg/L NO2- may compromise fish survival especially during long shipment abroad.

Effect of diflubenzuron on immature stages of Haematobia irritans (L.) (Diptera: Muscidae) in Uberlândia, State of Minas Gerais, Brazil

Horn fly immatures were raised in media containing different concentrations of diflubenzuron in order to verify their susceptibility to this insect growth regulator (IGR). The 50% and 95% lethal concentrations of diflubenzuron for the population (LC50, LC95) were determined as well as the effect of this IGR on the different immature horn fly stages. The tests were performed using the progeny of adults collected in the field. The immatures were maintained in a growth chamber at 25.0 ± 0.5ºC and 12-12 h photoperiod. IGR concentrations of 300 ppb, 100 ppb and 50 ppb were lethal for 100% of the sample. Pupae malformation occurred in the breeding media containing different diflubenzuron concentrations. Values for LC50 , LC95 (± 95% fiducial limits) and the slope of the regression line were respectively, 25.521 ± 1.981 ppb, 34.650 ± 2.001 ppb and 12.720 ± 1.096. The third larval instar was more sensitive to the sub-lethal concentration of the product than the first and second ones were. The results indicate that this IGR can be an important tool for controlling horn fly populations as well as for managing horn fly resistance to conventional insecticides against Haematobia irritans in Uberlândia, State of Minas Gerais.

Embryonic development of human lice: rearing conditions and susceptibility to spinosad

The embryonic development of human lice was evaluated according to the changes in the morphology of the embryo observed through the transparent chorion. Based on ocular and appendage development, three stages of embryogenesis were established: early, medium, and late. Influence of temperature and relative humidity (RH) on the laboratory rearing of Pediculus humanus capitis eggs was assessed. The optimal ranges for temperature and RH were 27-31°C and 45-75%. The susceptibility of human louse eggs to insecticide spinosad (a macrocyclic lactone) was assessed by immersion method. The results showed similar susceptibility to spinosad in early, medium, and late stages of head lice eggs. In addition, this study showed similar susceptibility of head and body lice eggs to spinosad, an insecticide that has not been used as pediculicide in Argentina (lethal concentration 50: 0.01%).

Current resistance status to temephos in Aedes aegypti from different regions of Argentina

In Argentina, more than 25,000 cases of dengue were reported in the summer of 2009, even in provinces where the disease was formerly absent. We analysed the susceptibility levels to the larvicide temephos in seven populations of Aedes aegypti, the primary vector of dengue, collected during summer 2007/2008, using the susceptible Rockefeller strain as a control. Although no control failures were observed during the experiment, a majority of the lethal concentration and resistance ratio values indicate an incipient resistance. An integrative program to monitor the resistance of Ae. aegypti to insecticides is needed in the country.

Comparison of the insecticide susceptibilities of laboratory strains of Aedes aegypti and Aedes albopictus

A susceptible strain of Aedes albopictus derived from the Gainesville strain (Florida, USA) was established in our laboratory. The larvicidal efficacies of the neurotoxic insecticides temephos, permethrin and the pure cis and trans-permethrin isomers and the microbial insecticide Bacillus thuringiensis israelensis (Bti) against Ae. albopictus were estimated and compared to a susceptible strain of Aedes aegypti. The larvicidal effect of insect growth regulator pyriproxyfen was also evaluated in both mosquito strains. The median lethal concentration/median emergency inhibition values for Ae. aegypti and Ae. albopictus, respectively, were: temephos, 3.058 and 6.632 ppb, permethrin, 3.143 and 4.933 ppb, cis-permethrin, 4.457 and 10.068 ppb, trans-permethrin, 1.510 and 3.883 ppb, Bti, 0.655 and 0.880 ppb and pyriproxyfen, 0.00774 and 0.01642 ppb. Ae. albopictus was more tolerant than Ae. aegypti to all six larvicides evaluated. The order of susceptibility for Ae. aegypti was pyriproxyfen > Bti > trans-permethrin > temephos > permethrin > cis-permethrin and for Ae. albopictus was pyriproxyfen > Bti > trans-permethrin > permethrin > temephos > cis-permethrin. Because both species can be found together in common urban, suburban and rural breeding sites, the results of this work provide baseline data on the susceptibility of Ae. albopictus to insecticides commonly used for controlling Ae. aegypti in the field.