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  • 1
    ISSN: 1574-6941
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A mesocosm experiment was performed to study the influence of nutrients on activity and diversity of bacterial assemblages from the Mediterranean Sea. Changes in the diversity of the predominant bacterial populations were monitored by DGGE fingerprinting of PCR products derived from 16S rRNA encoding genes. Fluctuations in the diversity of the most active populations was inferred by performing the DGGE fingerprinting on the basis of the cellular rRNA after reverse transcription and PCR amplification. DNA-derived DGGE patterns obtained from duplicate control and nutrient-enriched mesocosms showed differences in the development of the bacterial communities between control and nutrient-enriched experimental mesocosms. Multidimensional scaling analysis of the DNA-derived DGGE fingerprints indicated that duplicate treatments were reproducible. DNA- and RNA-derived DGGE fingerprints of bacterial assemblages changed over time, showing that the composition of the bacterial assemblages, as well as the most active bacterial populations changed during different phases of the incubation. Sequences of predominant DGGE bands in RNA-derived patterns were similar to 16S rRNA gene sequences of members of the α-, γ- and δ-Proteobacteria and of the Cytophaga-Flavobacterium-Bacteroides phylum (CFB). Bands corresponding to Ruegeria-like bacteria and members of the CFB became especially dominant during the course of incubation, suggesting that these populations were important contributors to bacterial production and activity in the post-grazing phase of the experiment.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1574-6941
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Small subunit rDNA clone libraries were generated from amplified DNA of bacterioplankton taken at different time points from a mesocosm containing eutrophied Mediterranean seawater and made eutrophic by the addition of N and P. Analysis of 96 partial sequences indicated that 22% of the clones formed four clusters which showed the highest sequence similarity with the 16S rDNA sequence of Alteromonas macleodii DSM 6062T. A fifth cluster, comprising 31% of the clone sequences is moderately related to the A. macleodii sequence. Similarity between the almost complete sequences of two representatives of clone clusters 1, 2 and 3 and A. macleodii ranged between 97.7 and 98.1%. Four oligonucleotide probes, representing four clone clusters, were developed on the basis of partial clone sequences. Dot blot hybridization with PCR-amplified 16S rDNA from 739 clones revealed that 24% of clones belong to one of these clusters. Dot blot hybridization between the four probes and PCR-amplified 16S rDNA from 128 strains isolated from the mesocosm identified 21% of the isolates possessing the probe target region. While probes GP-1 and GP-4 unambiguously identified 0.8 and 4.0% of the strains, respectively, probes GP-2 and GP-3 showed cross hybridization with 16% of the strains. Analysis of the probe target region of the 16S rDNA of one of the isolates indeed demonstrated the presence of double peaks in the relevant region of the sequence which is indicative of microheterogeneity at the rrn operon level. Although some of the diversity can be attributed to intra-strain variation, the data indicate that the phylogenetic diversity of A. macleodii is higher than represented by the type strain of this species.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1574-6941
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Quantitative and qualitative changes in bacterial communities from the Mediterranean Sea were compared in duplicate batch mesocosms with or without addition of inorganic nutrients. Methods including traditional microbial ecology techniques, molecular biology and flow cytometry were combined to determine abundances, production, cell size, activity, culturability and taxonomic diversity of bacterial cells. Addition of nutrients and confinement resulted in an increase of bacterial densities which were rapidly controlled by protozoan grazing. Changes in bacterial activity and morphology were observed during the growth phase of bacteria and under grazing pressure. The proportion of medium-size and culturable cells increased during the growth phase. These cells were preferentially consumed by grazers resulting in a strong limitation of bacterial production. As a consequence of the grazing pressure, large cells were produced and contributed to the remaining bacterial productivity after grazing. Grazing had an effect on the taxonomic composition of bacterial communities by preferentially eliminating γ-Proteobacteria, α-Proteobacteria were preserved. It seems that some species from the genera Ruegeria and Cytophaga may have developed defence strategies to escape predation.
    Type of Medium: Electronic Resource
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