Digestion of Yeasts and Beta-1,3-Glucanases in Mosquito Larvae: Physiological and Biochemical Considerations

PLoS One. 2016 Mar 23;11(3):e0151403. doi: 10.1371/journal.pone.0151403. eCollection 2016.

Abstract

Aedes aegypti larvae ingest several kinds of microorganisms. In spite of studies regarding mosquito digestion, little is known about the nutritional utilization of ingested cells by larvae. We investigated the effects of using yeasts as the sole nutrient source for A. aegypti larvae. We also assessed the role of beta-1,3-glucanases in digestion of live yeast cells. Beta-1,3-glucanases are enzymes which hydrolyze the cell wall beta-1,3-glucan polyssacharide. Larvae were fed with cat food (controls), live or autoclaved Saccharomyces cerevisiae cells and larval weight, time for pupation and adult emergence, larval and pupal mortality were measured. The presence of S. cerevisiae cells inside the larval gut was demonstrated by light microscopy. Beta-1,3-glucanase was measured in dissected larval samples. Viability assays were performed with live yeast cells and larval gut homogenates, with or without addition of competing beta-1,3-glucan. A. aegypti larvae fed with yeast cells were heavier at the 4th instar and showed complete development with normal mortality rates. Yeast cells were efficiently ingested by larvae and quickly killed (10% death in 2 h, 100% in 48 h). Larvae showed beta-1,3-glucanase in head, gut and rest of body. Gut beta-1,3-glucanase was not derived from ingested yeast cells. Gut and rest of body activity was not affected by the yeast diet, but head homogenates showed a lower activity in animals fed with autoclaved S. cerevisiae cells. The enzymatic lysis of live S. cerevisiae cells was demonstrated using gut homogenates, and this activity was abolished when excess beta-1,3-glucan was added to assays. These results show that live yeast cells are efficiently ingested and hydrolyzed by A. aegypti larvae, which are able to fully-develop on a diet based exclusively on these organisms. Beta-1,3-glucanase seems to be essential for yeast lytic activity of A. aegypti larvae, which possess significant amounts of these enzyme in all parts investigated.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aedes / growth & development*
  • Animals
  • Glucan 1,3-beta-Glucosidase / metabolism*
  • Larva / enzymology*
  • Saccharomyces cerevisiae / enzymology*

Substances

  • Glucan 1,3-beta-Glucosidase
  • beta-1,3-exoglucanase

Grants and funding

FAG was funded by Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ, http://www.faperj.br/), grant number E-26/110.760/2013, Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, http://www.cnpq.br/), grant number 573959/2008-0) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, http://www.capes.gov.br/), grant number 1273/2011. RJD was funded by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, http://www.cnpq.br/), grant number 400607/2012-3. HMD was funded by funded by the Science Without Borders Program by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq Young Talent Fellowship, http://www.cnpq.br/), grant number 400584/2012-3). RSS was funded by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES Parasitology program, http://www.capes.gov.br/), grant number 1273/2011. RJD, VMD and FAG are staff members from their respective departments, and were funded by the National Institute of Science and Technology for Molecular Entomology (CNPq grant number 573959/2008-0) and by Science Without Borders Program (Special Visitant Researcher Fellowship, CNPq number 400607/2012-3). FAG was funded by FIOCRUZ, CAPES, CNPq and FAPERJ. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.