The use of toluidine blue for in situ detection of micro-organisms in foods

The use of toluidine blue to stain cryosections of food samples to detect sites of microbial growth was examined. The method successfully detected colonies and single cells of both yeasts and bacteria at magnifications of x 400 or below in the majority of foods studied. The method demonstrates great potential for studying the micro-environments in which micro-organisms grow.     

Plasmid profiles as indicators of the source of contamination of Staphylococcus aureus endemic within poultry processing plants.

A total of 530 strains of Staphylococcus aureus were isolated from the defeathering machinery of a chicken processing plant and from neck skin samples of carcasses at different stages of processing in two visits 4 weeks apart. Eleven different plasmid profiles were detected in the isolates, eight being common to both visits. The plasmid profiles of the strains forming the majority of the population on the freshly slaughtered birds were rarely present in the strains isolated from the pluckers (except at the entry to the first plucker) and were present in only a small proportion of the strains isolated from carcasses after plucking. However, the profiles from the strains isolated from the pluckers on both visits were different from those forming the majority of the population on the incoming birds but formed the major part of the carcass flora after plucking, suggesting that such strains were endemic. These strains were found as a small proportion of the isolates made from the incoming birds, suggesting that this was the route by which the endemic strains were introduced into the plant. Such endemic strains exhibited a clumping growth, even in liquid shake culture, which may have made it easier for them to become established on the pluckers and to resist cleaning and disinfection. This clumping phenotype was correlated with the presence of a 7.5-megadalton plasmid.     

The benzodiazepine receptor ligand, methyl beta-carboline-3-carboxylate, is both sedative and proconvulsant in chicks

Certain pharmacological properties of methyl beta-carboline-3-carboxylate (beta-CCM), a benzodiazepine receptor ligand, have been investigated in chicks. Although beta-CCM has been established previously as an "inverse agonist" of benzodiazepine receptors in rodents, having effects opposite to those of benzodiazepines in a variety of tests, in chicks this compound had a different pharmacological profile. Firstly, in contrast to the overt convulsant action of beta-CCM in other species, beta-CCM (0.05-40 mg/kg) did not produce convulsions by itself in chicks, but it was only proconvulsant. Secondly and most surprisingly, beta-CCM, like diazepam, produced in chicks a sedation which could be blocked by the benzodiazepine receptor antagonist Ro 15-1788. Thus it appears that beta-CCM can function both as an agonist and as an inverse agonist in this animal.     

Dependence of secretion and assembly of type 1 fimbrial subunits of Escherichia coli on normal protein export.

The export of fimbrial subunits was found to be diminished at the restrictive temperature in a strain bearing a secA(Ts) mutation. Likewise, export was inhibited in a strain harboring a malE-lacZ protein fusion upon induction of hybrid protein synthesis. Both conditions resulted in the accumulation of a precursor protein ca. 2,000 daltons larger than the mature fimbrial subunit.     

O2 delivery to contracting muscle during hypoxic or CO hypoxia

The consequences of a decreased O2 supply to a contracting canine gastrocnemius muscle preparation were investigated during two forms of hypoxia: hypoxic hypoxia (HH) (n = 6) and CO hypoxia (COH) (n = 6). Muscle O2 uptake, blood flow, O2 extraction, and developed tension were measured at rest and at 1 twitch/s isometric contractions in normoxia and in hypoxia. No differences were observed between the two groups at rest. During contractions and hypoxia, however, O2 uptake decreased from the normoxic level in the COH group but not in the HH group. Blood flow increased in both groups during hypoxia, but more so in the COH group. O2 extraction increased further with hypoxia (P less than 0.05) during concentrations in the HH group but actually fell (P less than 0.05) in the COH group. The O2 uptake limitation during COH and contractions was associated with a lesser O2 extraction. The leftward shift in the oxyhemoglobin dissociation curve during COH may have impeded tissue O2 extraction. Other factors, however, such as decreased myoglobin function or perfusion heterogeneity must have contributed to the inability to utilize the O2 reserve more fully.     

Climate and the evolution of brachycephalization

Significant associations of cranial shape for 82 ethnic groups and seven climate variables are described. Variation among current populations is partially attributed to cold adaptation throughout the Pleistocene. Application of data files tabulated by the authors is described for a number of problems. Temporal distribution of 115 specimens indicates a geometric trend (CI = 76.7-1.96 log time X 10(3) ). Cranial indices are summarized within alternative taxonomic models and between climatic ecotypes. Evidence supports the hypothesis of cold adaptation among "Classic" Neandertals. Limitations of the thermodynamic model are discussed. It is probable that a decrease of the cranial index occurs from the Middle to Upper Paleolithic. During the Holocene, the index increases under all climatic conditions.     

Aerobic respiratory activity of the yeastlike phase of Sporotrichum schenckii as affected by high hydrogen ion concentrations

Summary Six strains ofSporotrichum schenckii were studied in regard to the effect of hydrogen ion concentration on certain aspects of the aerobic respiratory activity of the yeastlike phase.Optimal oxygen uptake of endogenous respiration occurred at pH 2.0, although no effects were observed on the oxygen-carbon dioxide exchange ratio (respiratory quotient) at this pH value when compared to the R.Q. obtained at pH 7.0. Endogenous respiratory activity at both pH 2 and 7 was markedly sensitive to the presence of certain respiratory inhibitors.Optimal respiratory activity using glucose as substrate occurred at pH 7.0 .On the other hand, oxidation of pyruvate as substrate proceeded at significant rates only at pH values below pH 4.0. With decreasing hydrogen ion concentration, accumulation of this organic acid occurred when glucose was employed as substrate. With the exception of acetate, none of the organic acid respiratory intermediates were found to stimulate respiration.The results reported herein suggest that the respiratory activity of the yeastlike phase ofS. schenckii differs in several respects from that observed for the yeastlike phases ofHistoplasma capsulatum andBlastomyces dermatitidis.From the Research Laboratory, Veterans Administration Hospital, Kansas City, Missouri, and The Department of Microbiology, Kansas University Medical Center, Kansas City, Kansas.Supported in part by USPHS Grant AI 03485.     

The ultrastructural localization of acid phosphatase,alkaline phosphatase and adenosine triphosphatase in induced goitres of Xenopus laevis Daudin tadpoles

Summary Tadpoles of Xenopus laevis were maintained in solutions of potassium perchlorate, potassium thiocyanate, thiourea and propyl-thiouracil (0.005% w/v). Metamorphosis was inhibited and within a short time large well-vascularized goitres developed. The localization of certain phosphatases at the ultrastructural level in these goitres is described and the possible roles played by these enzymes is discussed. Acid phosphatase was localized in follicle epithelial cells in dense droplets thought to be lysosomes or cytolysomes. Alkaline phosphatase and a Mg⁺⁺-activated ATP-ase were found on the outer surfaces of the membranes of follicular cells and on the walls of capillary blood vessels.This work was carried out during the tenure by one of us (R.C.) of a Medical Research Council Scholarship and forms part of a programme of research in amphibian thyroid physiology supported by a grant from the Medical Research Council, which we gratefully acknowledge.