A survey for Babesia microti in rodents was conducted at six sites within Grand Teton National Park, Wyoming. Blood and spleen smears, hematocrits, and reticulocyte counts were made on all of the animals to evaluate parameters for the diagnosis of babesiosis. Ticks were removed for identification. Of 257 Microtus montanus, 103 were infected with B. microti. In addition, five of 12 Microtus pennsylvanicus and one of three Arvicola richardsoni were parasitized by B. microti. Peromyscus maniculatus (n = 40) were not infected. Concurrent infections by Hepatozoon sp., Trypanosoma sp., and the bacterium, Grahamella sp., were noted in blood smears from a number of M. montanus. Splenomegaly and reticulocytosis were significant parameters associated with babesiosis while decreased hematocrit was not. Ticks removed from the voles were identified as Ixodes eastoni and were the probable vectors of the B. microti.

In searching for an invertebrate host for Hepatozoon sp. infecting the montane vole (Microtus montanus), we collected fleas, ticks, and mites from live-trapped voles and searched squash preparations for Hepatozoon oocysts. From 1989 through 1996, we identified six species of fleas in Grand Teton National Park: Megabothris abantis, Megabothris asio megacolpus, Aetheca wagneri, Peromyscopsylla selenis, Peromyscopsylla. hesperomys, and Hystrichopsylla dippiei dippiei. We found Hepatozoon oocysts only in M. abantis; we found no oocysts in mites or ticks. We conclude that M. abantis is an invertebrate host of Hepatozoon sp. and is likely to be the definitive host for theHepatozoon spp. of M. montanus.

RapidlyFive strains of a rapidly growing, orange, scotochromogenic Mycobacterium species were isolated from a marine sponge. Although they displayed similarities to M. aurum and M. parafortuitum, they proved to belong to a proposed new species, Mycobacterium poriferae sp. nov. (ATCC 35087). We distinguished the type strain of M. poriferae from that of M. aurum by its pattern of carbon sources used, acid production from carbon sources, and amidases, by its ability to tolerate 5% sodium chloride, its failure to use benzamide as sole nitrogen source, and its use of L-serine as sole carbon and nitrogen source. We distinguished the type strain of M. poriferae from that of M. parafortuitum by its pattern of carbon sources used, acid production from carbon sources, and amidases, by its ability to tolerate 5% sodium chloride, its failure to grow at 42"C, its strong pigmentation, its failure to reduce nitrate, its failure to tolerate 0.0250% hydroxylamine hydrochloride, its ability to use L-serine as dual carbon and nitrogen source, and its failure to use acetamide as dual carbon and nitrogen source. The pattern of mycolic acid derivatives produced by acid methanolysis of whole organisms was that of the group typified by M. avium and which contains a-mycolates, ketomycolates, and wax ester derivatives.The demosponge Halichondria bowerbanki contains the bicyclic aryl carotenoid isorenieratene (lo), a pigment found as a major carotenoid in certain Mycobacterium species and in the brown members of the Chlorobiaceae. Since a previous study showed that the carotenoid content in explants from H. bowerbanki increased with time in the dark as well as in the light (3, a nonphotosynthetic organism could have been a contributing source of carotenoids extracted from the sponge. We now report finding a new mycobacterial species as a result of studies designed to isolate and identify pigmented mycobacteria associated with H . bowerbanki. MATERIALS AND METHODSOrganisms. Living specimens of the marine sponge H. bowerbanki, known commonly as the "crumb-of-bread sponge," were obtained from Gulf Specimen Company (Panacea, Fla.). They were used for the preparation of cell suspensions within 24 h of their arrival.Primary isolation. A cell suspension was prepared from cut sponge pieces by mixing 1 volume of sponge pieces cut to approximately 3 by 3 mm and enough sterile seawater to make 3 volumes. The mixture was reduced to a suspension of cells and cell fragments by shearing with a Waring blender operated at low speed for 1 min. All equipment was sterile, and aseptic techniques were used. This suspension then was used to inoculate by loop slants of Middlebrook 7-H-11 medium (1). Slants were incubated either in air or 5% COz in air at 20 to 30°C for 4 to 12 weeks. Colonies of orange acid-fast organisms were purified by subculture on Tsukamura modified Sauton medium containing 5% (wt/vol) NaCl or medium supplemented with 1.0% (wt/vol) mannitol, 1.0% (wt/vol) sucrose, or 0.1 M monoethanolamine.Maintenance of strains. Pure stock cultures of Myco...

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1. The incorporation of [2-(14)C]mevalonic acid by Chloropseudomonas ethylica strain 2K into chlorobactene was studied. 2. Oxidative degradation of chlorobactene of constant specific radioactivity produced labelled benzenecarboxylic acids and indicated that the benzene ring originates from mevalonic acid. 3. Decarboxylation studies demonstrated a stereospecific methyl migration in the formation of the 1,2,5-trimethylphenyl group of chlorobactene. The migrating methyl group was derived from the C-3' position of mevalonic acid.