Sphingolipid-Containing Outer Membrane Vesicles Serve as a Delivery Vehicle To Limit Macrophage Immune Response to Porphyromonas gingivalis.

Sphingolipids (SLs) are essential structural components of mammalian cell membranes. Our group recently determined that the oral anaerobe Porphyromonas gingivalis delivers its SLs to host cells and that the ability of P. gingivalis to synthesize SLs limits the elicited host inflammatory response during cellular infection. As P. gingivalis robustly produces outer membrane vesicles (OMVs), we hypothesized that OMVs serve as a delivery vehicle for SLs, that the SL status of the OMVs may impact cargo loading to OMVs, and that SL-containing OMVs limit elicited host inflammation similar to that observed by direct bacterial challenge. Transwell cell culture experiments determined that in comparison to the parent strain W83, the SL-null mutant elicited a hyperinflammatory immune response from THP-1 macrophage-like cells with elevated tumor necrosis factor alpha (TNF-α), interleukin 1ß (IL-1ß), and IL-6. Targeted assessment of Toll-like receptors (TLRs) identified elevated expression of TLR2, unchanged TLR4, and elevated expression of the adaptor molecules MyD88 and TRIF (Toll/IL-1 receptor domain-containing adaptor-inducing beta interferon) by SL-null P. gingivalis No significant differences in gingipain activity were observed in our infection models, and both strains produced OMVs of similar sizes. Using comparative two-dimensional gel electrophoresis, we identified differences in the protein cargo of the OMVs between parent and SL-null strain. Importantly, use of purified OMVs recapitulated the cellular inflammatory response observed in the transwell system with whole bacteria. These findings provide new insights into the role of SLs in P. gingivalis OMV cargo assembly and expand our understanding of SL-OMVs as bacterial structures that modulate the host inflammatory response.

Cooking copepods: The survival of cyclopoid copepods (Crustacea: Copepoda) in simulated provisioned water containers and implications for the Guinea Worm Eradication Program in Chad, Africa.

INTRODUCTION: The global Guinea Worm Eradication Program has reduced numbers of human infections of Guinea worm disease (dracunculiasis) to 49 cases in four countries. However, infections of domestic animals (dogs and cats) have recently been recognized and are increasing. Typically, Guinea worm (Dracunculus medinensis) transmission occurs via the ingestion of copepods from water. Despite several interventions, including tethering of dogs while worms emerge, the number of infected dogs continue to increase. One hypothesis is that dogs could be infected through the ingestion of copepods in provisioned water. OBJECTIVES: The purpose of this study was to determine whether copepods can survive in water containers under typical Chadian temperatures. METHODS: Four container types (plastic, glass, gourd, and metal) were seeded with copepods and exposed to simulated Chadian temperatures. RESULTS: All copepods in the metal containers died within 4 h. Conversely, after 8 h live copepods were still present in plastic, glass, and gourd containers. CONCLUSIONS: If provisioned water is provided to potential hosts of D. medinensis, metal containers create the most inhospitable environment for copepods. Plastic containers have little effect on copepod mortality. The use of metal containers for water provisions could be a useful tool assisting with the interruption of D. medinensis transmission among dogs.