Alkaline phosphatase activity was assayed by two procedures in duodenal homogenates from specific pathogen-free (SPF) mice of various ages, adult germfree mice, adult ex-germ-free mice colonized with an indigenous microflora from the SPF mice, and adult ex-germ-free mice monocontaminated with a Lactobacillus sp. indigenous to the SPF mice. In the SPF neonates, the activity remained at low levels until at least 12 days of age, increased to high levels at 20 days of age, and then fell to adult levels between the early neonatal and later high levels. In the germ-free mice, the activity levels were significantly higher than the levels in SPF mice at any age. In contrast, in the ex-germ-free animals, colonized by an entire indigenous microflora, the values fell within the range for adult SPF animals. In the ex-germ-free mice colonized only by the Lactobacillus sp., the activity levels were intermediate between the values for germ-free and SPF mice. These findings show that the indigenous microbial flora influences considerably the intestinal alkaline phosphatase activity in the mouse.

Alkaline phosphatase activity was assayed in duodenal homogenates or extracts from adult specific pathogen-free (SPF) and germfree mice and gnotobiotic mice monoassociated with a Lactobacillus sp., a Bacteroides sp., or a coliform strain indigenous to SPF mice. Activity levels of the enzyme were much higher in the preparations from germfree mice than in those from the SPF controls. In the gnotobiotes monoassociated either with a freshly isolated Lactobacillus sp. or a Bacteroides sp., the levels of alkaline phosphatase activity were intermediate between the values for germfree and SPF mice. By contrast, in the gnotobiotes monoassociated with a coliform strain, alkaline phosphatase activity remained at high germfree levels. Butanol extracts of duodenal tissue from SPF mice, germfree mice, and exgermfree mice associated with an indigenous microflora from SPF mice (conventionalized) were subjected to acrylamide gel electrophoresis. A stain for alkaline phosphatase activity revealed three major bands in the gels prepared with extracts from SPF and conventionalized mice, but only two in the gels prepared with extracts from germfree mice. All three bands may have been present in the latter gels. One of the bands (the middle one) may have been obscured, however, by high activity in the slowest moving band. As determined by densitometric scanning, the slowest moving band had much higher activity in the preparations from germfree animals than in those from SPF or conventionalized mice. These findings suggest that the indigenous microbial flora affects not only quantitatively, but also qualitatively, the activity of alkaline phosphatases in the mouse intestinal mucosa.

Two distinct spore appendage types of Clostridium bijermentans, a pinlike appendage and a tubular appendage, were studied by electron microscopy. The pinlike appendage is characterized by a shaft, about 100 A in diameter, which has a lobed caplike structure. The tubular appendage, 500 to 600 A in diameter, is characterized by a hirsute region consisting of small filaments or fibrils. Gross morphology and ultrastructural features of both types are described. Recently, we reported the occurrence of four distinct spore appendage types among 12 strains of Clostridium bifermentans examined (9). In that report, we characterized two of the appendage types, one a smooth tubular appendage (strain 9-SDH) and the other a featherlike appendage (strain 1A-SDH). In the present report, we characterize the remaining two appendage types, a pinlike appendage (strain U-11) and a hirsute tubular appendage (strain FDA-1). In addition, some ultrastructural features of the previously reported smooth tubular appendage of strain 9-SDH are described. MATERIALS AND METHODS The experimental approach and procedures used in the present investigation were identical with those detailed in a previous report (9), and thus only an abbreviated account of these methods is given here. Three strains of C. bifermenttanis were used. Strains U-li and FDA-i were received from Stanley M. Harmon, Food and Drug Administration,

Four distinct spore appendage types were detected in an electron microscope survey of 12 strains of Clostridium bifermentans. A smooth tubular appendage and a feather-like appendage are described in detail. In addition, hirsute tubular appendages and small pin-like appendages are depicted. Spores of four strains apparently lack appendages.

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