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TETRACYCLINE FLUORESCENCE IN PERMEABILITY STUDIES OF MEMBRANES AROUND INTRACELLULAR PARASITES.

Certain protozoa, bacteria, and viruses when phagocytosed by host cells become surrounded by an intracytoplasmic boundary. This membrane prevents the fluorescent antibiotic tetracycline from entering the parasites when it is added to the medium, since they show no fluorescence, whereas extracellular parasites are immediately visible. As soon as the host cell dies, the intracellular parasites also become visible. This indicates that the boundary probably is of host origin. This phenomenon provides a means for selective permeability studies of such boundaries. A similar exclusion of tetracycline from certain extracellular parasites is seen in the presence of whole serum.

Animals↗

ABNORMAL IMMUNE MECHANISM IN ALLOGENEIC RADIATION CHIMERAS.

Mice were subjected to x-rays (950 roentgens) and injected with isogeneic (isologous) or allogeneic (homnologous) bone marrow. Six to 8 months later these chimeras were injected with Salmonella typhi flagellar antigen, and the formation of antibodies resistant and sensitive to destruction by treatment in vitro with 2-mercaptoethanol was determined. The allogeneic chimeras showed almost normal amounts of serumn antibody after a third injection of antigen but a relative defect in their ability to synthesize antibody resistant to 2-mercaptoethanol. Apparently control of antibody formation becomes abnormal in the presence of the immunologic tolerance existing between the host and the foreign hematopoietic graft.

Animals↗

Fate of staphylococci and enteric microorganisms introduced into slurry of frozen pot pies.

A slurry was prepared from six frozen pot pies diluted 1:5 with distilled water, two chicken, two turkey, and two beef pies of different brands. This slurry formed a reference sample and was placed in sterilized jars, frozen, and used as needed throughout the experiments. A second slurry was prepared in a similar manner from a frozen beef pot pie and a chicken pot pie, and was used as a control in only one experiment. The total count of microorganisms and the number of coliforms, Escherichia coli, salmonellae, and coagulase-positive staphylococci per gram were determined. Samples of slurry were inoculated in decimal dilutions with one or more of the following: Salmonella typhimurium, E. coli, and a strain of staphylococcus that causes food poisoning. The natural flora was found to exert an inhibitory effect upon the growth of the added microorganisms after incubation at 35 C for 18 hr. The inhibitory effect on growth was in part due to pH. The predominating organism isolated from the natural flora after incubation was a lactobacillus, which, when added in mixture with the test organisms in sterilized slurry, did not exert the profound inhibitory effect observed in the case of the natural flora. Some factors which may be concerned with the inhibition were investigated.

Animals↗

EFFECT OF WATER EXTRACTS OF CAROB PODS, TANNIC ACID, AND THEIR DERIVATIVES ON THE MORPHOLOGY AND GROWTH OF MICROORGANISMS.

The effect of aqueous extracts of carob (Ceratonia siliqua) pods, gallotannic acid, gallic acid, and catechol on several microorganisms was studied. Carob pod extract and tannic acid showed a strong antimicrobial activity toward some cellulolytic bacteria. On the basis of tannin content, to which antimicrobial effect was related, carob pod extracts inhibited Cellvibrio fulvus and Clostridium cellulosolvens at 15 mug/ml, Sporocytophaga myxococcoides at 45 mug/ml, and Bacillus subtilis at 75 mug/ml. The inhibiting concentrations for tannic acid were found to be 12, 10, 45, and 30 mug/ml, respectively. Gallic acid and catechol were much less effective. Tannic acid and the tannin fraction of carob extract exerted both bacteriostatic and bactericidal effects on C. fulvus. Respiration of C. fulvus in the presence of bactericidal concentrations of tannic acid or tannin fraction of carob extract was inhibited less than 30%. A partial formation of "protoplasts" by C. fulvus was obtained after 2 hr of incubation in a growth medium to which 20% sucrose, 0.15% MgSO(4).7H(2)O, and 10 to 50 mug/ml of tannic acid or 500mug/ml of penicillin, or both, had been added. Tannic acid and the tannin fraction of carob extract protected C. fulvus from metabolic lysis in sucrose solution. Although the growth of other microorganisms tested was only slightly affected, the morphology of some of them was drastically changed in the presence of subinhibitory concentrations of carob pod extracts of tannic acid. It is suggested that the site of action of tannins on sensitive microorganisms is primarily the cell envelope.

Bacillus subtilis↗

ISOLATION OF SALMONELLAE BY SELECTIVE MOTILITY SYSTEMS.

Salmonellae were isolated from naturally infected swine and human feces by means of selective migration through semisolid enrichment (SM) media in modified "U" tubes. Comparative studies showed that recovery of Salmonella by SM techniques was equal or superior to that of standard procedures employed in two routine diagnostic laboratories. Primary Salmonella isolations from SM enrichment were relatively free from normal fecal bacteria and often pure. The SM method required only 15% of the media and correspondingly less time and skill than the usual methods for isolating Salmonella.

Animals↗