Attached and unattached bacterial communities in a 120-meter corehole in an acidic, crystalline rock aquifer.

The bacteria colonizing geologic core sections (attached) were contrasted with those found suspended in the groundwater (unattached) by examining the microbiology of 16 depth-paired core and groundwater samples using a suite of culture-independent and culture-dependent analyses. One hundred twenty-two meters was continuously cored from a buried chalcopyrite ore hosted in a biotite-quartz-monzonite porphyry at the Mineral Park Mine near Kingman, Ariz. Every fourth 1.5-m core was acquired using microbiologically defensible methods, and these core sections were aseptically processed for characterization of the attached bacteria. Groundwater samples containing unattached bacteria were collected from the uncased corehole at depth intervals corresponding to the individual cores using an inflatable straddle packer sampler. The groundwater was acidic (pH 2.8 to 5.0), with low levels of dissolved oxygen and high concentrations of sulfate and metals, including ferrous iron. Total numbers of attached cells were less than 10(5) cells g of core material(-1) while unattached cells numbered about 10(5) cells ml of groundwater(-1). Attached and unattached acidophilic heterotrophs were observed throughout the depth profile. In contrast, acidophilic chemolithotrophs were not found attached to the rock but were commonly observed in the groundwater. Attached communities were composed of low numbers (<40 CFU g(-1)) of neutrophilic heterotrophs that exhibited a high degree of morphologic diversity, while unattached communities contained higher numbers (ca. 10(3) CFU ml(-1)) of neutrophilic heterotrophs of limited diversity. Sulfate-reducing bacteria were restricted to the deepest samples of both core and groundwater. 16S ribosomal DNA sequence analysis of attached, acidophilic isolates indicated that organisms closely related to heterotrophic, acidophilic mesophiles such as Acidiphilium organovorum and, surprisingly, to the moderately thermophilic Alicyclobacillus acidocaldarius were present. The results indicate that viable (but possibly inactive) microorganisms were present in the buried ore and that there was substantial distinction in biomass and physiological capabilities between attached and unattached populations.

Evaluation of a fluorescent lectin-based staining technique for some acidophilic mining bacteria.

A fluorescence-labeled wheat germ agglutinin staining technique (R. K. Sizemore et al., Appl. Environ. Microbiol. 56:2245-2247, 1990) was modified and found to be effective for staining gram-positive, acidophilic mining bacteria. Bacteria identified by others as being gram positive through 16S rRNA sequence analyses, yet clustering near the divergence of that group, stained weakly. Gram-negative bacteria did not stain. Background staining of environmental samples was negligible, and pyrite and soil particles in the samples did not interfere with the staining procedure.

The arsenical resistance operon of IncN plasmid R46.

The arsenical resistance operon of the IncN plasmid R46 consists of 4696 bp and starts with predicted transcriptional control and initiation signals, followed by five genes, arsD, arsA, and arsC. The corresponding Escherichia coli chromosomal ars operon and two staphylococcal ars operons lack arsA and arsD genes. The R46 system contains only the second known versions of arsA and arsD, after those of plasmid R773. Western blot analysis identified the R46 proteins using antibodies against R773 ArsA, ArsD and ArsR.

Characterization of a new bacteriophage which infects bacteria of the genus Acidiphilium.

A novel bacteriophage, termed phi AC1, that infects strains of the genus Acidiphilium (acidophilic, heterotrophic, aerobic, Gram-negative eubacteria) most commonly isolated from acidic mine drainage environments, has been discovered and several of its properties have been determined. This is the first report of a bacteriophage infecting such cells. The virion has a lambdoid morphology and is larger than lambda, as shown by electron microscopy and sucrose gradient centrifugation. The sedimentation coefficient of the virion is approximately 615S. The nucleic acid of phi Ac1 is dsDNA, approximately 102 kb in length. Several experimental results show that phi Ac1 is a temperate phage. The plaques are turbid, and most cells isolated from plaques produced on sensitive cells by filter-sterilized phage preparations contain the phage and are resistant to further phage infection. Southern blot analysis shows that phi Ac1 prophage DNA is integrated into the bacterial genome during the temperature growth phase.

Selenate reduction by bacteria from a selenium-rich environment.

Samples collected from Kesterson Reservoir were screened for bacterial presence and selenate reduction capability. Selenate concentrations of 100 mg/liter were not toxic to indigenous bacteria. Of the 44 samples collected, 20 possessed microbial populations capable of reducing selenate. Reduction was observed in 4% of the water samples, 92% of the sediment samples, and 100% of the soil samples. Microbial reduction of 100 mg of selenate per liter was complete within 1 week of incubation. Up to 75 mg of selenate per liter was reduced beyond selenite to an insoluble red precipitate. Data collected indicate that indigenous bacteria have a significant role in the biogeochemical cycling of selenium.

Stillbirth. A humanistic response.

A predictable pattern of grief follows every perinatal death. Because of their lack of training in the area of death and bereavement, physicians have traditionally avoided acknowledging the grief process in patients who have experienced such a loss. Obstetricians must begin to recognize the appropriate intervention that needs to be instituted in the delivery room and postpartum period to help facilitate the normal grieving process. Physicians must also acknowledge and accept their own feelings of grief and helplessness following a perinatal death.