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L A Amado

Publications and source records attributed to L A Amado.

3 recordsLinked to original sources

Biological variation between two Brazilian geographical isolates of Echinostoma paraensei.

The biological behaviour and morphometric data from two allopatric isolates of Echinostoma paraensei (Rio Bonito - RB and Sumidouro - SU) collected from naturally infected Nectomys squamipes from two secluded Atlantic Forest fragments were studied. Mice that had been experimentally infected with ten encysted metacercariae of each isolate were monitored weekly in two trials to analyse worm burden and the kinetics of worm distribution along the intestine. The total number of uterine eggs, wet weights and measurements of the worms and body, acetabulum, testes and ovaries were also analysed. The RB isolate showed a higher worm burden, 7.7+/-0.8, and a longer life span, 16 weeks, compared to a worm burden of 5.8+/-1.1 and life span of 9 weeks for the SU isolate. Worms of the RB isolate were clustered in the duodenum and in the bile duct while the SU isolate worms were dispersed along the small intestine of infected mice. Both isolates developed similarly as regards morphometric data and wet weight, although the total number of uterine eggs was greater in RB. The degree of intraspecific variation observed in the worm distribution along the intestine, worm burden and life span raises questions regarding the use of these criteria for species differentiation. These findings suggest that variation in biological parameters found between the E. paraensei isolates could result from geographical isolation and, in particular, the environmental conditions of transmission. Further studies on E. paraensei polulations from different forest fragments will contribute towards an understanding of the speciation of this parasite.

Animals↗

In situ enzyme immunoassay for titration of a Brazilian hepatitis A virus strain (HAF-203).

Hepatitis A virus (HAV) replicates relatively slowly in cell culture without a cytopathic effect, a fact that limits the use of tissue culture assays. The radioimmunofocus assay is the standard method for HAV titration, although it is labor intensive and requires the use of radioisotopes. A simple, rapid and objective infectivity assay based on an in situ enzyme immunoassay (EIA) is described here for a Brazilian cell culture-adapted HAV strain (HAF-203). The assay uses a peroxidase-labeled polyclonal antibody to fixed monolayers as an indicator of infection. EIA may be completed within 7 days using serial 5-fold dilutions of the virus, yielding a titer of 5.024 log 50% tissue culture infective dose (TCID50)/ml for HAF-203. This technique had a detection limit of 1.1 log TCID50/ml and the specificity was demonstrated by detecting no reaction on the columns of uninfected wells. The reproducibility (with intra- and inter-assay coefficients of variation ranging from 1.9 to 3.8% and from 3.5 to 9.9%, respectively) and quantitation of the assay were demonstrated by close agreement in virus infectivity titers among different assays of the same amount of virus and between assays of different amounts of virus. Furthermore, this assay does not require the use of radiolabeled antibodies. We describe here an efficient EIA that is highly reproducible and that could be used to monitor HAV growth in cell culture and to determine the quantity of HAV antigen needed for diagnostic assays. This is the first report of the infectious titer of the Brazilian cell culture-adapted HAV strain (HAF-203).

Hepatitis A virus↗