title: Morpho-functional characterization and esterase patterns of the midgut of Tribolium castaneum Herbst, 1797 (Coleoptera: Tenebrionidae) parasitized by Gregarina cuneata (Apicomplexa: Eugregarinidae).
abstract: Tribolium castaneum (Coleoptera: Tenebrionidae) is a common pest of stored grains and byproducts and is normally infected by Gregarina cuneata (Apicomplexa: Eugregarinidae). The life cycle of this parasite includes the sporozoite, trophozoite, gamont, gametocyte, and oocyst stages, which occur between the epithelium and lumen of the host's midgut. This study aims to describe the morphofunctional alterations in the midgut and determine the esterase patterns in T. castaneum when parasitized by gregarines. To achieve this purpose, midguts of adult insects were isolated, processed, and analysed using light and electron microscopy. We determined total protein content, amylase activity, and the expression and related activities of the esterases by using polyacrylamide gel electrophoresis (PAGE). The midgut of T. castaneum is formed by digestive, regenerative, and endocrine cells. The effects of parasitism on the digestive cells are severe, because the gregarines remain attached to these cells to absorb all the nutrients they need throughout their development. In these cells, the most common alterations observed include expansion and fragmentation of the rough endoplasmic reticulum, development of the smooth endoplasmic reticulum, changes in mitochondrial cristae, cytoplasmic vacuolization, formation of myelin structures, spherites, large intercellular spaces, autophagic vesicles, expansion of the basal labyrinth, and cytoplasmic protrusions. Deposits of glycogen granules were also observed. Amylase activity was reduced in parasitized insects. Regenerative cells were found in disorganized crypts and did not differentiate into new cells, thus, compromising the restoration of the damaged epithelium. Though few morphological alterations were observed in the endocrine cells, results suggest that the synthesis and/or release of hormones might be impaired. Nine esterases (EST-1 to 9) were identified in the midgut of T. castaneum and were expressed in varying levels in response to parasitism. Two additional isoforms of esterases were exclusively identified in the parasitized insects. The results of this study suggest that gregarines alter the morphology and physiology of the midgut. The changes may result in nutritional depletion and the impairment of other physiological processes, such as reproduction and development of the host. Thus, further studies are needed to uncover the possibility of utilizing gregarines as biological controllers of the insect pest population.
authors.affiliation: Departamento de Biotecnologia, Genética e Biologia Celular, Universidade Estadual de Maringá (UEM), Paraná, Brazil. Electronic address: adrianasinopolis@hotmail.com.
authors.forename: Adriana A Sinópolis
authors.initials: AA
authors.lastname: Gigliolli
authors.affiliation: Departamento de Biotecnologia, Genética e Biologia Celular, Universidade Estadual de Maringá (UEM), Paraná, Brazil. Electronic address: aslapenta@uol.com.br.
authors.forename: Ana Silva
authors.initials: AS
authors.lastname: Lapenta
authors.affiliation: Departamento de Biotecnologia, Genética e Biologia Celular, Universidade Estadual de Maringá (UEM), Paraná, Brazil. Electronic address: mccrtakasusuki@uem.br.
authors.forename: Maria Claudia Colla
authors.initials: MC
authors.lastname: Ruvolo-Takasusuki
authors.affiliation: Departamento de Bioquímica, Universidade Estadual de Maringá (UEM), Paraná, Brazil. Electronic address: josi_abrahao@hotmail.com.
authors.forename: Josielle
authors.initials: J
authors.lastname: Abrahão
authors.affiliation: Departamento de Biotecnologia, Genética e Biologia Celular, Universidade Estadual de Maringá (UEM), Paraná, Brazil. Electronic address: hconte@gmail.com.
authors.forename: Hélio
authors.initials: H
authors.lastname: Conte
mesh_terms: D000818:Animals; D016782:Apicomplexa / Q000254:growth & development*; D004591:Electrophoresis, Polyacrylamide Gel; D004950:Esterases / Q000032:analysis*; D041981:Gastrointestinal Tract / Q000033:anatomy & histology / Q000201:enzymology; D019476:Insect Proteins / Q000032:analysis; D008853:Microscopy; D014231:Tribolium / Q000201:enzymology* / Q000469:parasitology*
chemical_list: D019476:Insect Proteins; D004950:Esterases
keywords: Isoenzymes; Midgut; Morphology; Parasites; Stored grain pests