Prevotella copri variants among a single host diverge in sphingolipid production.

Sphingolipids serve as vital structural and signaling components of the cell membranes in both eukaryotes and prokaryotes. Within the gut microbiome, Bacteroides species have been identified as major producers of sphingolipids, and Bacteroides-produced sphingolipids have been shown to be modulators of host immune and metabolic functions. While Bacteroides species are a prominent feature of the gut microbiomes of populations living in industrialized countries, Prevotella copri, a member of the same phyla, albeit a different family, is the dominant feature across the remainder of the global population, although their sphingolipid-producing capabilities have not been as thoroughly investigated. To fill this gap, we examined the genomes of over 60 diverse isolates of P. copri and identified several key enzymes involved in sphingolipid synthesis in P. copri. Combining bioorthogonal labeling and liquid chromatography-mass spectrometry (LC-MS) based lipidomics, we functionally characterized the first step in P. copri de novo sphingolipid synthesis in addition to profiling the sphingolipidomes of P. copri strains, identifying key enzymes that may play roles in producing a diverse set of P. copri sphingolipids. Given the limited genetic engineering tools amenable for use in P. copri, our approach takes advantage of comparative genomics and phenotypic profiling to explore sphingolipid production in these understudied, yet highly prevalent, organisms.IMPORTANCESphingolipids are important signaling molecules for maintaining metabolic and immune homeostasis in the host. These lipids are also produced by gut commensals, most notably by Bacteroides species. Despite the global prevalence of Prevotella copri in gut microbiomes of individuals, little is known about the types of sphingolipids they produce and whether they are similar in composition and structure to those produced by Bacteroides. Given the varied associations of P. copri with diverse sphingolipid-related health outcomes, such as rheumatoid arthritis and glucose intolerance, it is important to first characterize the specific sphingolipids produced by individual strains of P. copri and to identify the genes involved in their pathways of production. This characterization of P. copri-derived sphingolipids provides further insight into how bacterial sphingolipid production can serve as a mechanism for microbial modulation of host phenotypes.

Effects of Nitrate and Conductivity on Embryo-Larval Fathead Minnows.

Nitrate concentrations have been rising in surface waters over the last century and now frequently exceed drinking water standards and environmental safety benchmarks globally. Health-wise, these trends are concerning because nitrate has been shown to disrupt endocrine function and developmental outcomes. The present study investigated potential sublethal effects of nitrate on developing fathead minnows. Fish were exposed from fertilization through 21 days postfertilization (dpf) to environmentally relevant concentrations of nitrate (0, 2, 5, 10, 25, or 100 mg/L NO3 -N as NaNO3 ). Nitrate effects on hatch timing, heart rate and rhythm at 3 dpf, growth through 21 dpf, swim bladder inflation timing and size, scoliosis, pericardial edema, and mortality were assessed. Because adding NaNO3 increases water conductivity, two conductivity controls were included to match the ionic strength of the 10- and 100-mg/L NO3 -N treatments. Increasing nitrate delayed posterior swim bladder (PSB) inflation in a dose-dependent manner, with possible inhibition of anterior swim bladder (ASB) inflation at higher doses, although nitrate did not affect swim bladder size. Conversely, nitrate did not affect hatch timing or cardiac endpoints at 3 dpf or induce pericardial edema or scoliosis, although there was a noted brood effect on these latter defects. As was observed with increasing nitrate, higher ion concentrations in the conductivity controls caused dose-dependent increases in fish body size at 21 dpf. Increased ionic strength also hastened ASB inflation independently of nitrate. As in other published studies, the observed delay in PSB inflation suggests that nitrate disrupts the thyroid axis and warrants further investigation. In addition, the present study supports the need for conductivity controls in nitrate toxicity studies to distinguish nitrate-specific effects. Environ Toxicol Chem 2023;42:1529-1541. Published 2023. This article is a U.S. Government work and is in the public domain in the USA.