Susceptibility of Phytomonas serpens to calpain inhibitors in vitro: interference on the proliferation, ultrastructure, cysteine peptidase expression and interaction with the invertebrate host

A pleiotropic response to the calpain inhibitor MDL28170 was detected in the tomato parasite Phytomonas serpens. Ultrastructural studies revealed that MDL28170 caused mitochondrial swelling, shortening of flagellum and disruption of trans Golgi network. This effect was correlated to the inhibition in processing of cruzipain-like molecules, which presented an increase in expression paralleled by decreased proteolytic activity. Concomitantly, a calcium-dependent cysteine peptidase was detected in the parasite extract, the activity of which was repressed by pre-incubation of parasites with MDL28170. Flow cytometry and Western blotting analyses revealed the differential expression of calpain-like proteins (CALPs) in response to the pre-incubation of parasites with the MDL28170, and confocal fluorescence microscopy confirmed their surface location. The interaction of promastigotes with explanted salivary glands of the insect Oncopeltus fasciatus was reduced when parasites were pre-treated with MDL28170, which was correlated to reduced levels of surface cruzipain-like and gp63-like molecules. Treatment of parasites with anti-Drosophila melanogaster (Dm) calpain antibody also decreased the adhesion process. Additionally, parasites recovered from the interaction process presented higher levels of surface cruzipain-like and gp63-like molecules, with similar levels of CALPs cross-reactive to anti-Dm-calpain antibody. The results confirm the importance of exploring the use of calpain inhibitors in studying parasites’ physiology.

Relative size of gills of Cloeodes jaragua Salles & Lugo-Ortiz, 2003 (Ephemeroptera, Baetidae) on pool and riffle areas of streams at the Atlantic Rainforest / Tamanho relativo das brânquias de Cloeodes jaragua Salles & Lugo-Ortiz, 2003 (Ephemeroptera, Baetidae) em áreas de corredeiras e remansos em córregos na Mata Atlântica

Physical environment of streams present many challenging factors to the organisms living under its influence. Water speed, one of the most important factors affecting such environments, has local influence on substrata, temperature, and oxygen levels. It acts determining species distributions and morphological and behavioral adaptations of organisms. The aim of this study was to assess the relative size of gills of Cloeodes jaragua on two mesohabitats: lentic environment (river dam) and lotic environment (fast flowing area of a stream) in Parque Nacional da Serra dos Órgãos (RJ, Brazil). Twenty individuals were collected on each environment and width of the head and gills were taken. Lentic environment showed nymphs with broader and more constant measurements than the lotic environment. The former, despite being composed of nymphs on different stages of development, presented individuals with narrower gills. Both, head width and environment type explained gills size, showing that streams habitat conditions are important in shaping the morphology of these animals.

O ambiente físico dos rios apresenta diversos desafios para os organismos que vivem sob sua influência. A velocidade da água, um dos fatores mais importantes a atuar nesses ambientes, tem influência local no substrato, temperatura e oxigenação da água. A força da correnteza age determinando a distribuição das espécies e as adaptações morfológicas e comportamentais dos organismos. O objetivo desse estudo foi abordar o tamanho relativo das brânquias de Cloeodes jaragua em dois meso-habitats: um ambiente lêntico (represa) e um ambiente lótico (áreas de correnteza) no Parque Nacional da Serra dos Órgãos - RJ. Um total de 20 indivíduos foi coletado em cada tipo de ambiente e as larguras da cabeça e brânquias foram medidas. O ambiente lêntico apresentou ninfas com medidas mais largas e mais constantes que o ambiente lótico. Este último, apesar de ser composto por ninfas em diferentes estágios de desenvolvimento, apresentou indivíduos com menores larguras de brânquias. Ambos, largura da cabeça e tipo de ambiente, explicaram o tamanho das brânquias, mostrando as condições dos habitats dos rios como importantes modeladores da morfologia desses animais.