The potential of microdialysis to monitor organic and inorganic nitrogen compounds in soil

Plant nitrogen (N) acquisition is strongly controlled by the concentration of available inorganic and organic N in the soil solution and by biogeochemical processes in the rhizosphere. However, until now it was hardly possible to reliably estimate plant-available N in soil microsites. Here, a novel microdialysis approach based on passive diffusion sampling is presented and compared qualitatively and quantitatively with lysimeter and soil extraction techniques when analyzing two contrasting boreal soils. Further, preliminary dialysis membrane calibration issues for sampling plant-available N compounds are discussed. Due to its miniaturized design microdialysis was shown to be a suitable tool for continuous sampling of ammonium, nitrate and free amino acids from the soil solution with only minimal disturbance of the soil structure. Microdialysis proved to be outstanding regarding the possible spatial (<0.5 mm) and temporal (<30 min) resolution of soil solution N chemistry. The different methods for soil N sampling resulted in significantly different results. In lysimeter and soil extraction samples, nitrate and ammonium were found at the highest concentrations, while results from microdialysis revealed that the pool of plant-available amino acids was contributing most to the total N pool tested. Application of a standard N solution to the tested soils led to an immediate peak of recovery via the microdialysis probes followed by a rapid decrease due to the formation of a depletion zone at the probe surfaces. Therefore, this relatively new technique will not only provide essential data on diffusion rates of a variety of N compounds in the soil but might be used for monitoring quantitative and qualitative changes in plant-available N in soil microsites such as the rhizosphere.     

Structures of the bacterial ribosome in classical and hybrid states of tRNA binding

During protein synthesis, the ribosome controls the movement of tRNA and mRNA by means of large-scale structural rearrangements. We describe structures of the intact bacterial ribosome from Escherichia coli that reveal how the ribosome binds tRNA in two functionally distinct states, determined to a resolution of ~3.2 angstroms by means of x-ray crystallography. One state positions tRNA in the peptidyl-tRNA binding site. The second, a fully rotated state, is stabilized by ribosome recycling factor and binds tRNA in a highly bent conformation in a hybrid peptidyl/exit site. The structures help to explain how the ratchet-like motion of the two ribosomal subunits contributes to the mechanisms of translocation, termination, and ribosome recycling.     

Latitudinal differences in the duration of development of Neocalanus plumchrus copepodites

Quasi‐synoptic sampling along a transect from Alaska to California carried out in spring and summer 2000 using Continuous Plankton Recorders reveals that the abundant calanoid copepod Neocalanus plumchrus (Marukawa) reaches the surface at approximately the same time across the region. However, monthly sampling also reveals that the timing of peak biomass (when 50% of the population consists of copepodites at stage CV) occurs about 5 weeks earlier at the southernmost part of its range than at the northernmost part, with intermediate areas having intermediate timings. Surface water temperature differed by about 4°C between the warmer south and the cooler north, and we suggest that such a difference would reduce development duration by about 3 weeks. If food supply in the north is more conducive to lipid accumulation, as seems possible from phytoplankton biomass data, then the use of the differences in timing of peak N. plumchrus biomass in each area as a measure of the differences in development time may not be accurate. We find that the estimate of the time of peak biomass varying by about 5 weeks between north and south may be an exaggeration of the actual difference in the development duration. Temperature is probably the most important factor governing the shorter development duration of N. plumchrus in southern latitudes.     

Application of Relaxed Eddy Accumulation Technique to Quantify Hg0 Fluxes Over Modified Soil Surfaces

Fluxes of mercury from a waste repository and from an agricultural field amended with sewage sludge were measured. The measurements were performed by utilising a Relaxed Eddy Accumulation (REA) system synchronously coupled to automated mercury cold vapour atomic fluorescence analysers (CVAFA). The waste repository was severely contaminated and fluxes of between 0.16μg m^?2 h^?1 and 29μg m^?2 h^?1 could be measured. Fluxes at the slightly contaminated agricultural field measured between 8 ng m^?2 h^?1 and 1.9μg m^?2 h^?1. The results showed that the REA-CVAFA system could be used for measurements above both severely-and slightly contaminated soils. The measurements at the chlor-alkali plant showed that temperature was the most important parameter controlling the release of mercury at this site. At the agricultural field, the measurements indicated that both solar radiation and temperature may have influenced the emissions. Most of the mercury added to the field was lost to the atmosphere during the summer season.     

Expression of interferon gamma in the brain of cats with natural Borna disease virus infection

Borna disease virus (BDV) is a neurotropic, negative-stranded RNA virus, which causes a non-suppurative meningoencephalomyelitis in a wide range of animals. In cats, BDV infection leads to staggering disease. In spite of a vigorous immune response the virus persists in the central nervous system (CNS) in both experimentally and naturally infected animals. Since the CNS is vulnerable to cytotoxic effects mediated via NK-cells and cytotoxic T-cells, other non-cytolytic mechanisms such as the interferon (IFN) system is favourable for viral clearance. In this study, IFN-γ expression in the brain of cats with clinical signs of staggering disease (N=12) was compared to the expression in cats with no signs of this disease (N=7) by quantitative RT-PCR. The IFN-γ expression was normalised against the expression of three reference genes (HPRT, RPS7, YWHAZ). Cats with staggering disease had significantly higher expression of IFN-γ compared to the control cats (p-value ≤ 0.001). There was no significant difference of the IFN-γ expression in BDV-positive (N=7) and -negative (N=5) cats having clinical signs of staggering disease. However, as BDV-RNA still could be detected, despite an intense IFN-γ expression, BDV needs to have mechanisms to evade this antiviral immune response of the host, to be able to persist.     

Investigation into association and expression of PLCz and COX-2 as candidate genes for boar sperm quality and fertility

Phospholipase C zeta (PLCz) and cyclooxygenase isoenzyme type 2 (COX-2) are important in spermatogenesis, but their effect has not yet confirmed in pigs. Therefore, this study was aimed to analyse their association with sperm quality and fertility and to identify the mRNA and protein expression in boars reproductive tissues. DNA samples from 231 Pietrain (PI) and 109 Pietrain × Hampshire (PIHA) pigs with records of sperm quality [sperm concentration (SCON), motility, semen volume, plasma droplet and abnormal spermatozoa rate] and fertility (non-return rate and number of piglet born alive) traits were available. A SNP in non-coding region of PLCz g.158 A > C was associated with SCON (p < 0.05) in PIHA population while the polymorphism of COX-2 g.68 G > A in 3' UTR was not associated with any traits. For mRNA and protein expression study, a total of six boars were divided into two groups with G-I and G-II, where G-I was characterized for relatively better sperm quality. Both genes expressed higher in reproductive tissues compared with non-reproductive tissues. Phospholipase C zeta mRNA expressed higher in testis (p < 0.01), all parts of epididymis and spermatozoa from G-I, while COX-2 expressed higher in testis (p < 0.05), head and body of epididymis (p < 0.01), and spermatozoa from G-II boar. Both proteins were localized in Leydig cells and spermatozoa. These results might shed light on roles of these genes in spermatogenesis as candidate for boar sperm quality and fertility, but still the lack of association across populations should be considered.     

Land cover influences on juvenile Rainbow Trout diet composition and condition in Lake Michigan tributaries

Watershed development may alter tributaries in ways that influence the growth and development of juvenile fish. For Rainbow Trout Oncorhynchus mykiss, the extent of land cover influence on the diet composition of stream‐residing juveniles is still an open but important question, particularly given their broad global distribution. Our study evaluated the effect of land cover on diet composition and condition metrics of juvenile Rainbow Trout in Lake Michigan tributaries. Juveniles were collected in the fall of 2014 and 2015 from 18 sampling locations across the Lake Michigan basin and included a wide variety of land cover types. Multivariate statistical procedures were used to quantitatively score sites along axes that maximized variation in land cover and diet composition. Subsequently, Spearman's rank correlations were used to test for significance of correlations between (a) land cover and diet composition, (b) land cover and fish condition, and (c) diet composition and fish condition. Our results indicated that land cover had an influence on the diet compositions of juvenile Rainbow Trout, but neither land cover nor diet composition had a significant influence on Rainbow Trout condition. These findings contrast with previously published work focusing on the growth responses of other fish species and provide insight into the plastic nature of Rainbow Trout. Further, we discuss potential ways in which fish may mitigate the effect of land cover changes as they are transmitted through stream food webs; however, the magnitude of the mitigation likely varies among fish species.     

Brazilian spotted fever: A spatial analysis of human cases and vectors in the state of São Paulo,Brazil

Brazilian spotted fever (BSF) is a highly lethal disease caused by Rickettsia spp. and is transmitted by ticks of the genus Amblyomma. Understanding the epidemiology of BSF in each region can help direct health surveillance actions. The objective of this study was to determine the spatial distribution of vectors and the incidence of BSF in the state of São Paulo. Spatial analysis included confirmed BSF cases between 2009 and 2019; occurrences were registered by municipalities of the state. There were 752 confirmed BSF cases, with incidences ranging between 0.01 and 10.37/10,000 inhabitants. Moran's Global Index for BSF was 0.20 (p = .001), which was indicative of spatial dependence. Moran's map located a cluster of 20 high priority municipalities for BSF and showed that Amblyomma sculptum is the vector involved in Rickettsia rickettsii transmission in these locations. Spatial analysis identified clusters formed by 47, 20 and seven significant municipalities for the presence of A. sculptum, Amblyomma aureolatum and Amblyomma ovale vectors, respectively. Surveillance and prevention actions are necessary in areas that are at high risk for BSF and in areas where the presence of vectors was significant.