Seroprevalence of Neospora caninum in dairy goats from Bahia, Brazil

Sera from 384 dairy goats in nine herds from Bahia State, Brazil, were assayed for Neospora caninum antibodies using an indirect immunofluorescent antibody test (IFAT). Animals were selected and divided in groups by age and breed. Antibodies against N. caninum were found in 15% (58/384) of goats. Serum titers ranged from 1:100 to 1:3200. There was no significant difference between the age groups. Seroprevalence was higher in Alpine breed (24%) than in Saanen (14%) and Nubian (3%) breeds. The 58 N. caninum-positive sera were also tested for Toxoplasma gondii and 71% of the samples reacted solely to N. caninum. These results indicate that N. caninum infection is common in goat herds in Bahia.     

De novo phosphatidylcholine synthesis is required for autophagosome membrane formation and maintenance during autophagy

ABSTRACT

Macroautophagy/autophagy can enable cancer cells to withstand cellular stress and maintain bioenergetic homeostasis by sequestering cellular components into newly formed double-membrane vesicles destined for lysosomal degradation, potentially affecting the efficacy of anti-cancer treatments. Using ¹³C-labeled choline and ¹³C-magnetic resonance spectroscopy and western blotting, we show increased de novo choline phospholipid (ChoPL) production and activation of PCYT1A (phosphate cytidylyltransferase 1, choline, alpha), the rate-limiting enzyme of phosphatidylcholine (PtdCho) synthesis, during autophagy. We also discovered that the loss of PCYT1A activity results in compromised autophagosome formation and maintenance in autophagic cells. Direct tracing of ChoPLs with fluorescence and immunogold labeling imaging revealed the incorporation of newly synthesized ChoPLs into autophagosomal membranes, endoplasmic reticulum (ER) and mitochondria during anticancer drug-induced autophagy. Significant increase in the colocalization of fluorescence signals from the newly synthesized ChoPLs and mCherry-MAP1LC3/LC3 (microtubule-associated protein 1 light chain 3) was also found on autophagosomes accumulating in cells treated with autophagy-modulating compounds. Interestingly, cells undergoing active autophagy had an altered ChoPL profile, with longer and more unsaturated fatty acid/alcohol chains detected. Our data suggest that de novo synthesis may be required to increase autophagosomal ChoPL content and alter its composition, together with replacing phospholipids consumed from other organelles during autophagosome formation and turnover. This addiction to de novo ChoPL synthesis and the critical role of PCYT1A may lead to development of agents targeting autophagy-induced drug resistance. In addition, fluorescence imaging of choline phospholipids could provide a useful way to visualize autophagosomes in cells and tissues.