Late Aquitanian mammals from Engehalde (Molasse Basin,Canton Bern,Switzerland)

The construction of the Neufeld tunnel in Bern city (2006–2008) led to the discovery of fossil mammals in the vicinity of the historical site of Engehalde. The study of the whole available sample led to the distinction of two mammal assemblages: (1) the historical level with the rhinocerotids Diaceratherium lemanense and D. aginense, and the ruminant Andegameryx cf. laugnacensis; (2) the Neufeld level with the artiodactyls Dremotherium feignouxi and Cainotherium sp., the carnivore Plesiogale angustifrons, and the glires Eucricetodon cf. aquitanicus, Peridyromys sp., and Prolagus vasconiensis. These two assemblages attest to a latest Aquitanian age (MN2b) for the last deposits of the ‘Lower Freshwater Molasse’ in the area. The palaeoecological analysis of the faunas indicates either the co-occurrence or the very short-termed succession of assorted terrestrial environments, ranging from forested habitats nearby water bodies or nearby steady rivers to open well-drained habitats within the hinterland.     

Capsular Serotyping of Streptococcus pneumoniae Using the Quellung Reaction

There are over 90 different capsular serotypes of Streptococcus pneumoniae (the pneumococcus). As well as being a tool for understanding pneumococcal epidemiology, capsular serotyping can provide useful information for vaccine efficacy and impact studies. The Quellung reaction is the gold standard method for pneumococcal capsular serotyping. The method involves testing a pneumococcal cell suspension with pooled and specific antisera directed against the capsular polysaccharide. The antigen-antibody reactions are observed microscopically. The protocol has three main steps: 1) preparation of a bacterial cell suspension, 2) mixing of cells and antisera on a glass slide, and 3) reading the Quellung reaction using a microscope. The Quellung reaction is reasonably simple to perform and can be applied wherever a suitable microscope and antisera are available.     

Assay of proteolytic enzymes by the fluorescence polarization technique.

Neuraminidase substrates of high specific activity (>300 μCi/μmol) were prepared by reduction of sialyllactose with NaB³H₄, followed by separation of the 2 → 3 and 2 → 6 isomers of [³H]sialyllactitol by paper chromatography. Hydrolysis of sialyllactitol by neuraminidase was monitored by measuring the radioactivity in the neutral reaction product, which was separated from the charged substrate by passage over a small anion exchange column. The assay was applied to the neuraminidase activity of cultured human skin fibroblasts. The K_m was found to be 1.1 mm for both substrates; the pH optimum, 4.0; the 2 → 3 isomer was hydrolyzed twice as fast as the 2 → 6. In several genetic disorders associated with neuraminidase deficiency, the activity toward both isomers was reduced almost completely (mucolipidoses I and II; Goldberg syndrome), or only partially (mucolipidosis III; adult myoclonus syndrome); however, the relative activity towards the two isomers remained approximately the same in all cases.