Comparative nitric oxide production by LPS-stimulated monocyte-derived macrophages from Ovis canadensis and Ovis aries

Bighorn sheep are more susceptible to respiratory infection by Mannheimia haemolytica than are domestic sheep. In response to bacterial challenge, macrophages produce a number of molecules that play key roles in the inflammatory response, including highly reactive nitrogen intermediates such as nitric oxide (NO). Supernatants from monocyte-derived macrophages cultured with M. haemolytica LPS were assayed for nitric oxide activity via measurement of the NO metabolite, nitrite. In response to LPS stimulation, bighorn sheep macrophages secreted significantly higher levels of NO compared to levels for non-stimulated macrophages. In contrast, levels of NO produced by domestic sheep macrophages in response to M. haemolytica LPS did not differ from levels detected in non-stimulated cell cultures. Nitrite levels detected in supernatants of LPS-stimulated bighorn macrophage cultures treated with an inducible nitric oxide synthase (INOS) inhibitor, N(G)-monomethyl-L-arginine, were similar to that observed in non-stimulated cultures indicating a role for the iNOS pathway.     

A single promoter inversion switches Photorhabdus between pathogenic and mutualistic states

Microbial populations stochastically generate variants with strikingly different properties, such as virulence or avirulence and antibiotic tolerance or sensitivity. Photorhabdus luminescens bacteria have a variable life history in which they alternate between pathogens to a wide variety of insects and mutualists to their specific host nematodes. Here, we show that the P. luminescens pathogenic variant (P form) switches to a smaller-cell variant (M form) to initiate mutualism in host nematode intestines. A stochastic promoter inversion causes the switch between the two distinct forms. M-form cells are much smaller (one-seventh the volume), slower growing, and less bioluminescent than P-form cells; they are also avirulent and produce fewer secondary metabolites. Observations of form switching by individual cells in nematodes revealed that the M form persisted in maternal nematode intestines, were the first cells to colonize infective juvenile (IJ) offspring, and then switched to P form in the IJ intestine, which armed these nematodes for the next cycle of insect infection.     

Response of soil microbial biomass and enzyme activities to the transient elevation of carbon dioxide in a semi-arid grassland

Although elevation of CO₂ has been reported to impact soil microbial functions, little information is available on the spatial and temporal variation of this effect. The objective of this study was to determine the microbial response in a northern Colorado shortgrass steppe to a 5-year elevation of atmospheric CO₂ as well as the reversibility of the microbial response during a period of several months after shutting off the CO₂ amendment. The experiment was comprised of nine experimental plots: three chambered plots maintained at ambient CO₂ levels of 360 μmol mol⁻¹ (ambient treatment), three chambered plots maintained at 720 μmol mol⁻¹ CO₂ (elevated treatment) and three unchambered plots of equal ground area used as controls to monitor the chamber effect.Elevated CO₂ induced mainly an increase of enzyme activities (protease, xylanase, invertase, alkaline phosphatase, arylsulfatase) in the upper 5 cm of the soil and did not change microbial biomass in the soil profile. Since rhizodeposition and newly formed roots enlarged the pool of easily available substrates mainly in the upper soil layers, enzyme regulation (production and activity) rather than shifts in microbial abundance was the driving factor for higher enzyme activities in the upper soil. Repeated soil sampling during the third to fifth year of the experiment revealed an enhancement of enzyme activities which varied in the range of 20-80%. Discriminant analysis including all microbiological properties revealed that the enzyme pattern in 1999 and 2000 was dominated by the CO₂ and chamber effect, while in 2001 the influence of elevated CO₂ increased and the chamber effect decreased.Although microbial biomass did not show any response to elevated CO₂ during the main experiment, a significant increase of soil microbial N was detected as a post-treatment effect probably due to lower nutrient (nitrogen) competition between microorganisms and plants in this N-limited ecosystem. Whereas most enzyme activities showed a significant post-CO₂ effect in spring 2002 (following the conclusion of CO₂ enrichment the previous autumn, 2001), selective depletion of substrates is speculated to be the cause for non-significant treatment effects of most enzyme activities later in summer and autumn, 2002. Therefore, additional belowground carbon input mainly entered the fast cycling carbon pool and contributed little to long-term carbon storage in the semi-arid grassland.     

Silicon identification in prosthesis-associated fibrous capsules

The use of correlated microscopic techniques, including the scanning electron microscopic modes of backscattered electron imaging and energy dispersive x-ray analysis, aid in defining the process of dispersion of silicon-containing material around silicone rubber (polydimethylsiloxane) prosthetic devices.