Spatial structure maintains diversity of pyocin inhibition in household Pseudomonas aeruginosa

Nearly all bacteria produce narrow-spectrum antibiotics called bacteriocins. Studies have shown that bacteriocins can mediate microbial interactions, but the mechanisms underlying patterns of inhibition are less well understood. We assembled a spatially structured collection of isolates of Pseudomonas aeruginosa from bathroom and kitchen sink drains in nine households. Growth inhibition of these P. aeruginosa by bacteriocins, known as pyocins in this species, was measured using pairwise inhibition assays. Carbon source usage of these isolates was measured, and genetic distance was estimated using multilocus sequencing. We found that as the distance between sites of isolation increased, there was a significantly higher probability of inhibition, and that pyocin inhibition and susceptibility vary greatly among isolates collected from different houses. We also detected support for other mechanisms influencing diversity: inhibition outcomes were influenced by the type of drain from which isolates were collected, and while we found no indication that carbon source utilization influences inhibition, inhibition was favoured at an intermediate genetic distance. Overall, these results suggest that the combined effects of dispersal limitation among sites and competitive exclusion within them maintain diversity in pyocin inhibition and susceptibility phenotypes, and that additional processes such as local adaptation and effects of phylogenetic distance could further contribute to spatial variability.     

Does aspartic acid racemization constrain the depth limit of the subsurface biosphere?

Previous studies of the subsurface biosphere have deduced average cellular doubling times of hundreds to thousands of years based upon geochemical models. We have directly constrained the in situ average cellular protein turnover or doubling times for metabolically active micro‐organisms based on cellular amino acid abundances, D/L values of cellular aspartic acid, and the in vivo aspartic acid racemization rate. Application of this method to planktonic microbial communities collected from deep fractures in South Africa yielded maximum cellular amino acid turnover times of ~89 years for 1 km depth and 27 °C and 1–2 years for 3 km depth and 54 °C. The latter turnover times are much shorter than previously estimated cellular turnover times based upon geochemical arguments. The aspartic acid racemization rate at higher temperatures yields cellular protein doubling times that are consistent with the survival times of hyperthermophilic strains and predicts that at temperatures of 85 °C, cells must replace proteins every couple of days to maintain enzymatic activity. Such a high maintenance requirement may be the principal limit on the abundance of living micro‐organisms in the deep, hot subsurface biosphere, as well as a potential limit on their activity. The measurement of the D/L of aspartic acid in biological samples is a potentially powerful tool for deep, fractured continental and oceanic crustal settings where geochemical models of carbon turnover times are poorly constrained. Experimental observations on the racemization rates of aspartic acid in living thermophiles and hyperthermophiles could test this hypothesis. The development of corrections for cell wall peptides and spores will be required, however, to improve the accuracy of these estimates for environmental samples.     

The effects of adrenalectomy on the sexual behaviour of castrated and intact BDF1 mice

Two experiments investigate the effects of adrenalectomy on the capacity of male mice of the BDF₁ genotype (C57B1/6Fa♀×DBA/2♂) to retain sexual behaviour following castration. Adrenalectomy is without effect and it is suggested that androgens can play no part in the maintenance of sexual behaviour in these castrates.     

Localized Organic Binding of Radioiodine in Hydra

Radioiodine, given as the iodide, localized in protein-bound form in the ectoderm of Hydra fusca and H. littoralis. The radioiodine was shown by autoradiography to be: (1) diffusely distributed in the ectoderm over the gonads and (2) precisely localized in the apical cytoplasm of cnidoblasts that carry stenotele nematocysts. The meaning of such precise cellular localization of iodoprotein in Hydra remains to be determined.     

STUDIES ON NITROGEN FIXATION BY ALNUS CRISPA

Root nodules of Alnus crispa (Ait.) Pursh were shown to possess a symbiotic nitrogen-fixing organism. The reduction of acetylene to ethylene, as measured by gas chromatography, was used to determine the presence of the nitrogen-fixing system. Ethylene production was measured at 5.1 μmoles/g excised nodule · hr for both field and greenhouse plants. The nodules were found to consist of short nubs usually clustered in masses up to 4 cm in diam. Microscopic examination of nodules revealed some cortical cells fully packed with spherical endophyte cells. The outer cortex and radiating arms of cells in the inner cortex remained uninfected. Nodules examined during the winter were found to be shrunken, with a random distribution of endophyte cells. Soil nitrogen measurements indicated that nitrogen fixation activity by A. crispa does not lead to an increase in soil nitrogen above levels in adjacent areas.     

A recombinant scFv/streptavidin-binding peptide fusion protein for the quantitative determination of the scorpion venom neurotoxin AahI.

We created a construct encoding a peptide known to mimic the binding properties of biotin fused to the carboxy-terminus of a scFv fragment that binds a scorpion toxin (AahI). This fusion protein was produced in the periplasm of bacteria and purified to homogeneity by single-step affinity chromatography on streptavidin-agarose with a yield close to 1 mg/l. DNA sequencing, dot blot and mass spectrometric analyses demonstrated the integrity of the soluble immunoconjugate. Fusion to the streptavidin-binding peptide did not affect the ability of the scFv to recognize its antigen with a high affinity (Kd = 2.3 x 10(-10) M). Similarly, the streptavidin-binding property was not impaired in the fusion protein. Thus, the immunoconjugate was bifunctional and had a low molecular mass of 28 kDa. This enabled us to develop rapid and sensitive immunoassays for the specific detection of the toxin AahI accurately to 0.6 ng/ml, opening up new perspectives for the diagnosis of envenomations.