The Saposin-Like Protein AplD Displays Pore-Forming Activity and Participates in Defense Against Bacterial Infection During a Multicellular Stage of Dictyostelium discoideum.

Due to their archaic life style and microbivor behavior, amoebae may represent a source of antimicrobial peptides and proteins. The amoebic protozoon Dictyostelium discoideum has been a model organism in cell biology for decades and has recently also been used for research on host-pathogen interactions and the evolution of innate immunity. In the genome of D. discoideum, genes can be identified that potentially allow the synthesis of a variety of antimicrobial proteins. However, at the protein level only very few antimicrobial proteins have been characterized that may interact directly with bacteria and help in fighting infection of D. discoideum with potential pathogens. Here, we focus on a large group of gene products that structurally belong to the saposin-like protein (SAPLIP) family and which members we named provisionally Apls (amoebapore-like peptides) according to their similarity to a comprehensively studied antimicrobial and cytotoxic pore-forming protein of the protozoan parasite Entamoeba histolytica. We focused on AplD because it is the only Apl gene that is reported to be primarily transcribed further during the multicellular stages such as the mobile slug stage. Upon knock-out (KO) of the gene, aplD- slugs became highly vulnerable to virulent Klebsiella pneumoniae. AplD- slugs harbored bacterial clumps in their interior and were unable to slough off the pathogen in their slime sheath. Re-expression of AplD in aplD- slugs rescued the susceptibility toward K. pneumoniae. The purified recombinant protein rAplD formed pores in liposomes and was also capable of permeabilizing the membrane of live Bacillus megaterium. We propose that the multifarious Apl family of D. discoideum comprises antimicrobial effector polypeptides that are instrumental to interact with bacteria and their phospholipid membranes. The variety of its members would allow a complementary and synergistic action against a variety of microbes, which the amoeba encounters in its environment.

Aedes aegypti population sampling using BG-Sentinel traps in north Queensland Australia: statistical considerations for trap deployment and sampling strategy.

BG-Sentinel mosquito traps were trialed as a tool for the rapid assessment (24-h collections) and routine monitoring (72-h collections) of adult Aedes aegypti L. populations in north Queensland. Analysis of Ae. aegypti collections using BG-Sentinels set in suburban Cairns for 24 h permitted the calculation of sample size for a range of precision levels. Clusters of houses with BG-Sentinels operating continuously for 15 d, with collections every 72 h, also permitted required sample size calculation. Evidence of Ae. aegypti spatial clustering at the house scale was revealed, with statistically significant effects detected for all collection days. Less variation was detected at each trap location, with only nine of 32 trap locations revealing significant clustering over time. Trap-out effects through continuous BG-Sentinel operation at a fixed location were absent. The findings support fixed position sampling at 72-h intervals for routine monitoring ofAe. aegypti populations in Cairns. Despite the relationship between collections of adult vectors and the incidence of disease remaining unknown, BG-Sentinel collections provide an alternative and less labor-intensive abundance measure for assessing risk of dengue virus transmission and success of dengue vector control programs.