Differential expression of brummer and levels of TAG in different developmental stages Aedes aegypti (Diptera: Culicidae), including fasted adults.

Lipid storage in the form of triacylglycerol (TAG) is essential for insect life, as it enables flight, development, and reproduction. The activity of the lipase brummer (bmm) has been shown to be essential to insects' homeostasis. The objective of this study was to evaluate how bmm expression occurs in Aedes aegypti larvae and adults, and to observe TAG levels during fasting in adult females. The bmm sequence was identified in A. aegypti and exhibited a patatin-like phospholipase domain reinforced by the presence of a catalytic dyad with serine and aspartate residues, revealing a high degree of similarity with other organisms. Bmm expression was differentiated in the larvae and adult fat body (FB) following TAG reserve dynamics. Bmm was expressed three times in larval stages L3, L4, and pupae compared with L1 and L2, which could indicate its role in the maturation of these insects. In the postemergence (PE) and post-blood meal (PBM) FB of adult insects, bmm expression varied over several days. PE adults showed a pronounced bmm increase from the third day onward compared with those not subjected to fasting. This was accompanied by a decrease in TAG from the third day onward, suggesting the participation of bmm. Six hours after blood feeding, TAG levels increased in mosquitos reared in the absence of sucrose, suggesting lipid accumulation to guarantee reproduction. Bmm responded positively to fasting, followed by TAG mobilization in adult FB. During the previtellogenic period, bmm levels responded to low TAG levels, unlike the PBM period.

Unveiling the oxidative metabolism of Achatina fulica (Mollusca: Gastropoda) experimentally infected to Angiostrongylus cantonensis (Nematoda: Metastrongylidae).

For the first time, alterations in the oxidative metabolism of Achatina fulica experimentally infected with different parasite loads of Angiostrongylus cantonensis were determined. For this, the hemolymph activities of lactate dehydrogenase (LDH) and hexokinase and the glucose concentrations in the hemolymph, as well as the polysaccharide reserves in the digestive gland and cephalopedal mass, were assessed. Additionally, the contents of some carboxylic acids in the hemolymph of infected and uninfected snails were determined by high-performance liquid chromatography (HPLC), permitting a better understanding of the alterations related to the host's oxidative metabolism. As the main results, activation of oxidative pathways, such as the glycolytic pathway, was demonstrated in response to the increase in the activity of hexokinase. This tendency was confirmed by the decrease in the contents of glucose in the hemolymph of parasitized snails, indicating that the infection by A. cantonensis alters the host's metabolism, and that these changes are strongly influenced by the parasite load. This metabolic scenario was accompanied by activation of the anaerobic fermentative metabolism, indicated not only by an increase in the activity of (LDH), but also by a reduction of the content of pyruvic acid and accumulation of lactic acid in the hemolymph of parasitized snails. In this circumstance, maintenance of the host's redox balance occurs through activation of the fermentative pathways, and LDH plays a central role in this process. Together, the results indicate that A. cantonensis infection induces activation of the anaerobic metabolism of A. fulica, characterized not only by the accumulation of lactic acid, but also by a reduction in the pyruvic acid and oxalic acid contents in the hemolymph of the infected snails.

Interaction of lipophorin with Rhodnius prolixus oocytes: biochemical properties and the importance of blood feeding

Lipophorin (Lp) is the main haemolymphatic lipoprotein in insects and transports lipids between different organs. In adult females, lipophorin delivers lipids to growing oocytes. In this study, the interaction of this lipoprotein with the ovaries of Rhodnius prolixus was characterised using an oocyte membrane preparation and purified radiolabelled Lp (125I-Lp). Lp-specific binding to the oocyte membrane reached equilibrium after 40-60 min and when 125I-Lp was incubated with increasing amounts of membrane protein, corresponding increases in Lp binding were observed. The specific binding of Lp to the membrane preparation was a saturable process, with a Kdof 7.1 ± 0.9 x 10-8M and a maximal binding capacity of 430 ± 40 ng 125I-Lp/µg of membrane protein. The binding was calcium independent and pH sensitive, reaching its maximum at pH 5.2-5.7. Suramin inhibited the binding interaction between Lp and the oocyte membranes, which was completely abolished at 0.5 mM suramin. The oocyte membrane preparation from R. prolixus also showed binding to Lp from Manduca sexta. When Lp was fluorescently labelled and injected into vitellogenic females, the level of Lp-oocyte binding was much higher in females that were fed whole blood than in those fed blood plasma.

Interaction of lipophorin with Rhodnius prolixus oocytes: biochemical properties and the importance of blood feeding.

Lipophorin (Lp) is the main haemolymphatic lipoprotein in insects and transports lipids between different organs. In adult females, lipophorin delivers lipids to growing oocytes. In this study, the interaction of this lipoprotein with the ovaries of Rhodnius prolixus was characterised using an oocyte membrane preparation and purified radiolabelled Lp (125I-Lp). Lp-specific binding to the oocyte membrane reached equilibrium after 40-60 min and when 125I-Lp was incubated with increasing amounts of membrane protein, corresponding increases in Lp binding were observed. The specific binding of Lp to the membrane preparation was a saturable process, with a K(d) of 7.1 ± 0.9 x 10-8M and a maximal binding capacity of 430 ± 40 ng 125I-Lp/µg of membrane protein. The binding was calcium independent and pH sensitive, reaching its maximum at pH 5.2-5.7. Suramin inhibited the binding interaction between Lp and the oocyte membranes, which was completely abolished at 0.5 mM suramin. The oocyte membrane preparation from R. prolixus also showed binding to Lp from Manduca sexta. When Lp was fluorescently labelled and injected into vitellogenic females, the level of Lp-oocyte binding was much higher in females that were fed whole blood than in those fed blood plasma.

Effects of infection by larvae of Angiostrongylus cantonensis (Nematoda, Metastrongylidae) on the lipid metabolism of the experimental intermediate host Biomphalaria glabrata (Mollusca: Gastropoda).

Experimental infection of Biomphalaria glabrata by Angiostrongylus cantonensis induces significant changes in the concentrations of triacylglycerol and cholesterol in the hemolymph and of neutral lipids in the digestive gonad-gland (DGG) complex of the host snail. In this study, snails were dissected after 1, 2, and 3 weeks of infection to collect the hemolymph and DGG and to measure the levels of cholesterol and triacylglycerol in the hemolymph and neutral lipid fractions in the tissues. The results show that infection by this nematode resulted in a significant decrease in the concentrations of both cholesterol and triacylglycerol in the hemolymph of B. glabrata during the parasite's initial ontogenic development period. This reduction indicates the possible use of these molecules by both parasite and host not only as energy substrates but also as structural factors required during development of the parasite's larval stages. In parallel, changes in the neutral lipid profile in the DGG and lipase activity of the infected snails were observed, indicating the importance of these molecules for successful infection.