RESUMO
Paracoccidioides brasiliensis and P. lutzii are fungi that cause paracoccidioidomycosis (PCM), the most prevalent systemic mycosis in South America. For serological diagnosis, although 43-kDa glycoprotein (gp43) is regarded as highly specific for PCM, the occurrence of false negative reactions in sera from patients infected with P. lutzii suggests that preparation with only one antigen is not recommended. Heat shock proteins are feasible alternatives as a second antigen because they are often highly immunogenic. In this study, we evaluated the usefulness of recombinant 60-kDa heat shock protein from P. brasiliensis (rPbHsp60) for the serological diagnosis of PCM. Using western blotting assay, we observed that 77.3% of the sera from PCM patients were positive to rPbHsp60, with 90.9% positivity to recombinant gp43 (rgp43). More importantly, sera from healthy subjects had 27% positivity to rPbHsp60 and none to rgp43. When rPbHsp60 was used in ELISA, we did not observe significant differences between the reactions with sera from PCM patients and healthy subjects, while the difference was clearly evident when the antigen was rgp43. Furthermore, rPbHsp60 was recognized by sera from patients with histoplasmosis, aspergillosis, sporotrichosis or tuberculosis in an ELISA test. These results show that rPbHsp60 is not a good antigen for PCM diagnosis.
Assuntos
Humanos , Antígenos de Fungos/sangue , Chaperonina 60/sangue , Proteínas Fúngicas/sangue , Paracoccidioides/imunologia , Paracoccidioidomicose/diagnóstico , Testes Sorológicos/métodos , Western Blotting , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Paracoccidioidomicose/sangue , Proteínas Recombinantes/sangue , Valores de Referência , Reprodutibilidade dos Testes , Estatísticas não ParamétricasRESUMO
In order to monitor the susceptibility of Boophilus microplus to acaricides and improve control measures, the effects of cypermethrin, deltamethrin and permethrin on larvae obtained in the city of Goiânia in the Brazilian state of Goiás were studied. Although these pyrethroids are already sold as acaricides, their cost-benefit efficiency has been questioned. Fasting 14-21 day-old larvae were immersed in solutions of the acaricides under test, maintained at 27±1° C, and relative humidity over 80%, and observed under the stereoscope within an apparatus originally designed for studying the non-parasitic phase of the tick life cycle. The observed toxicological effects were: excitability, repetitive motion, decreased motor ability, detachment, paralysis, knock-down and cuticular proliferation of liquids and gases. The materials used in the manufacture of this apparatus consisted of disposable Petri dishes, "organza" cloth and paraffin, none of which are toxic to tick larvae. Mean death rates after 24h were 76.3%, 87.5%, 77.6%, 91.2%, 86.2% and 100% for 25 and 50ppm deltamethrin, 150 and 300ppm cypermethrin and 1250 and 2500ppm permethrin, respectively. The ticks were resistant to commercial concentrations of deltamethrin and cypermethrin. Only 2500ppm permethrin produced the mortality recommended by the Brazilian Ministry of Agriculture.
Estudaram-se os efeitos de cipermetrina, deltametrina e permetrina sobre larvas de uma cepa de campo de Goiânia, com o objetivo de monitorar a susceptibilidade de Boophilus microplus para esses acaricidas e fomentar medidas de controle. Larvas em jejum com 14 a 21 dias, imersas em soluções desses piretróides, foram mantidas a 27±1°C e UR% > ou = 80% e observadas por 24h ao estereoscópio, contidas em dispositivo desenvolvido originalmente para estudos da fase não parasitária do ciclo evolutivo. O material utilizado em sua confecção, placa de petri descartável, tecido organza e parafina, não foi tóxico para as larvas. Os seguintes efeitos toxicológicos foram observados: excitabilidade, movimentação repetitiva, diminuição da capacidade locomotora, desprendimento, paralisia, knock-down e proliferação cuticular de líquidos e gases. A mortalidade média na 24ªh foi de 76,3%, 87,5%, 77,6%, 91,2%, 86,20%, e 100,0%, respectivamente, para deltametrina 25ppm e 50ppm, cipermetrina 150ppm e 300ppm e permetrina 1250ppm e 2500ppm. O ixodídeo foi resistente às concentrações comerciais de deltametrina e cipermetrina. Apenas permetrina 2500ppm provocou índice de mortalidade superior ao valor recomendado pelo Ministério da Agricultura para acaricidas.
RESUMO
Aiming at monitoring the susceptibility, increasing the control and evaluating the toxicological effects, of different concentrations of three pyrethroids were investigated in vitro against Rhipicephalus sanguineus larvae using concentrations formulated for cattle, dogs and household use. A total of 1,604 15-21 day-old fasting larvae, obtained by artificial infestation in dogs, was immersed in the solutions. Larvae were then maintained in BOD incubators inside experimental devices, constituted by disposable Petridishes, filter paper and paraffin, and observed on the stereomicroscope during a 24-hour period. The larvae have shown excitability, repetitive movements, decrease of locomotion, detachment, paralysis, knock-down and production of gases and liquid in the cuticle. Mortality after 24 hours was 86.9% (1250ppm) and 100.0% (2500ppm) for permethrin; 80.3% (150ppm) and 86.0% (300ppm) for cipermethrin; and for deltamethrin 68.2% (25ppm) and 78.0% (50ppm). Mortality was not observed in the control group.
Investigaram-se os efeitos toxicológicos in vitro de três piretróides em diferentes concentrações, formulados para uso pecuário, domiciliar e em cães, sobre larvas de R. sanguineus, objetivando monitorar sua suscetibilidade e fomentar seu controle. Utilizaram-se 1.604 larvas em jejum, com 15 a 21 dias, obtidas por infestação artificial em cão, imersas nas soluções testadas, contidas em dispositivos constituídos por placa de Petri descartável, papel filtro e parafina, mantidas em incubadoras do tipo BOD e observadas ao estereoscópio por 24h. Observaram-se excitabilidade, movimentação repetitiva, diminuição da locomoção, desprendimento, paralisia, "knock-down" e proliferação cuticular de gases e líquidos. A mortalidade na 24ª hora foi de 86,9%, 100,0%, 80,3%, 86,0%, 68,2% e 78,0%, respectivamente, para permetrina 1250ppm e 2500ppm, cipermetrina 150ppm e 300ppm, e deltametrina 25ppm e 50ppm. Não houve mortalidade no grupo controle.