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Fluorescent in situ hybridization (FISH) with rRNA -targeted oligonucleotide probes has become one of the major techniques in environmental microbiology, allowing rapid and reliable definition of prokaryotes and quantification of population sizes. The aim was to demonstrate the applicability of the FISH method to study bacterioplankton composition in North Mamry Lake, and to follow the dynamics of two populations of common bacteria. We analyzed the phylogenetic composition of free-living bacterioplankton assemblage using oligonucleotide probes specific for Bacteria as well as for β-Proteobacteria and Cytophaga-Flavobacterium groups. Up to 53% of bacteria detected with DA PI could be detected via FISH by applying the universal bacterial probe for domain Bacteria (Eub338). Percentage of Cytophaga-Flavobacterium cluster did not exceed 20%. Members of the β –Proteobacteria appeared to be the most abundant group.
The effects of inorganic nutrients (N, P) enrichment of mesotrophic lake water on changes in bacterial and protistan (heterotrophic nanoflagellates and ciliates) communities compositions were studied in the mesocosm experiment. Phosphorus (PO₄³⁻) and nitrogen (NH₄⁺) alone and in combination were added to three types of experimental mesocosms. Mesocosms results suggested that simultaneous addition of P and N stimulated phytoplankton growth and production rates of bacterial biomass its turnover rate. Strong positive correlations between chlorophyll a and bacterial secondary production rates suggested that bacteria were mainly controlled by organic substrates released in course of phytoplankton photosynthesis. Both nutrients increased distinctly protistan biomass and resulted in the shift in ciliate community composition from algivorous to large omnivorous species. The response of bacterial numbers and biomass to nutrients addition was less evident. However, intensive grazing caused their dynamic changes. Fluorescence in situ hybridization (FISH) revealed only small changes in bacterial taxonomic composition. There was an apparent shift in dominance from Cytophaga-Flavobacterium to the Alphaproteobacteria group in the mesocosm with simultaneous addition of P and N, which positively related to increased abundance of bacterivorous protists. Experiment demonstrated that inorganic N and P nutrients directly influenced the bottom-down control of microbial communities, which had a crucial effect on morphological diversity of bacteria.
Effects of mesotrophic lake water enrichment with organic phosphorus and nitrogen substrates (DNA and model protein, bovine serum albumin - BSA) on dynamics and diversity of natural microbial communities (bacteria, heterotrophic nanoflagellates, ciliates) were studied in mesocosm experiments. Simultaneous enrichment with DNA and BSA strongly increased the abundance and biomass of all studied groups of microorganisms and induced changes in their morphological and taxonomic structure. The increased participation of large heterotrophic nanoflagellates cells (larger than 10 μm) in their total numbers and shifts in taxonomic and trophic Structure of the ciliates, from algivorous to small bacterivorous, species were observed. Grazing caused changes in bacterial size distribution in all enriched mesocosms. Large (10-50 μm) filamentous bacteria significantly contributed to the total bacterial numbers and biomass. Pronounced increase in populations of (β- and γ-Proteobacteria was found in lake water enriched with organic P and N sources, whereas α-Pmteobacteria did not change markedly in the studied mesocosms. DNA additions stimulated the rates of bacterial secondary production. BSA shortened the rates of bacterial biomass turnover in lake water. Relatively high and constant (~ 30%) percentage contribution of active bacteria (MEM+) in two mesocosms enriched with DNA and DNA+BSA suggested the important role of nucleic acids as a source of phosphorus for bacterial growth, activity and production. Numerous and statistically significant correlations between bacteria and protists indicated the direct and selective predator-prey relationship.
Aquaporins (AQP) are hydrophobic integral membrane channel proteins that facilitated water transport across the plasma membrane. In this study, the reverse transcription real-time polymerase chain reaction (Real-Time RT-PCR) assay was used to determine the expression of genes encoding AQP1, AQP5 and AQP9 in porcine ovarian follicles, separated theca and granulosa cells of six experimental groups: early-luteal (days 2–4), mid-luteal (days 10–12 of the cycle, coinciding with a period of full active corpora lutea corresponding to the activity of corpora lutea in the period of pregnancy), late-luteal (days 14–16 of the cycle, coinciding with a period of luteal regression and development of a new cohort of follicles) and follicular group (days 18–20) of the oestrous cycle, as well as early implantation (days 14–16) and post-implantation, placentation group (days 30–32) of gestation. Significant differences in the AQP1, AQP5 and AQP9 genes expression between studied groups appeared only in the separated theca and granulosa cells. In the present study it is implied that the AQP1, 5 and 9 participate in the formation of follicular fluid and follicular development. These three examined AQPs appear to act interdependently, thereby maintaining tissue homeostasis.
In 2004-05 counts of the bacteria proteolytic, ammonifying, AOB and NOB, NO₃-N to NO₂-N reducing, denitrifying, and atmospheric nitrogen fixing (Azotobacter sp. and Clostridium pasteurianum) were analyzed along with temperature, pH, dissolved oxygen saturation, ammonia, nitrite and nitrate nitrogen in waters of the River Drwęca. The counts of the above groups of bacteria ranged within a few orders of magnitude depending on the physiological group and sampling site. The smallest were the counts of autotrophic nitrifying and atmospheric nitrogen fixing bacteria Azotobacter sp. (10² cfu·1 cm⁻³) and Clostridium pasteurianum (10² MPN·100 cm⁻³). The most numerous were ammonifying bacteria (10³ – 10⁶ MPN·100 cm⁻³). The quantitative occurrence of the bacteria in question varied in relation to the physical and chemical parameters of the water, which was evidenced by Spearman’s statistical analysis.
Water in the Drwęca River has been monitored in terms of differences in counts of bacteria which are active in transformations of nitrogen compounds depend on various man-made activity. In 2000–2001 samples of waters were assayed for counts of proteolytic, ammonifying, AOB, NOB, N-NO₃ to N-NO₂ reducing bacteria, denitrifying bacteria as well as anaerobic (Clostridium pasteurianum) and aerobic (Azotobacter sp.) atmospheric nitrogen binding bacteria, which differed within several orders (0–10⁵ cfu 1 cm⁻³ or MPN 100 cm⁻³) depending on the analysed physiological group of bacteria, sampling site or date. Statistically significant differences in counts of particular groups of microorganisms in the analysed water samples collected from the Drwęca River proved between the sampling dates only confirm that the river may be seasonally polluted by domestic sewage from towns and villages located in the river catchment or be associated with the cyclic nature of fisheries and/or agricultural production.
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