Protein digestion, anatomy and physiological conditions of the digestive tube of Lutzomyia longipalpis` larvae (Diptera: Psychodidae).
Vladimir Fazito do Vale; Marcos Horácio Pereira; Nelder de Figueiredo Gontijo.
Departamento de
Parasitologia, Instituto de Ciências Biológicas da UFMG; Belo Horizonte
- MG; CEP: 31 270901, Caixa Postal 486, e-mail: nelder@icb.ufmg.br.
The sand fly Lutzomyia longipalpis is the vector of Leishmania infantum (syn L. chagasi),
the etiologic agent of visceral leishmaniasis in the New World. Despite
its importance in public health, until now the internal anatomy of the
immature forms was not yet described and little is known about their
digestive processes. In nature, sand fly larvae feed on low nutrient
food like organic detritus in the soil, ingesting constantly a
relatively large quantity of material. The objective of this study was
to describe the anatomy of the digestive tube, the pH inside the lumen
as well as to investigate the proteases responsible for protein
digestion and their localization. The larvae posses a short gut with a
predominant well developed midgut. Ingestion of food containing
indicator dies permitted the measurement of the intestinal pH. A
pH gradient was observed varying from higher or equal to 9, in the
anterior midgut, to pH 6.5-7.0, in the posterior midgut. The
endoproteolytic enzymes are synthesized in the anterior midgut and are
able to digest azocasein over a large pH range. They are more effective
in high alkaline environments presenting an optimal activity in pH 11.
Studies with various inhibitors were performed and indicated that the
endoproteases are trypsin-like and chymotrypsin-like serine proteases.
These results were confirmed by using the substrates BApNA and BTpNA,
specific for trypsin and chymotrypsin respectively. Aminopeptidases
were also investigated using some p-Nitroaniline-derived substrates.
These enzymes are located in the posterior midgut, bound to the
membranes, presenting an optimal pH in the range of 6.5-7.5. The
results presented are in accordance with the proposal that proteins are
digested to small peptides in the anterior midgut inside the
endoperitrophic space and undergo subsequent digestion in the
ectoperitrophic space in the posterior midgut.
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