Biomedical subjects
Publications and source records attributed to T Michalowski.
Three strains of rumen treponemes were isolated and partially characterized. The strains differed significantly one from another in morphology, fermentation characteristics and plasmid profiles. Their genetic variability was assayed using DNA-based molecular approaches. Easily differentiated ARDRA (amplified ribosomal DNA restriction analysis) patterns indicated that the strains represent different bacterial species.
The diversity of methanogenic archaea associated with different species of ciliated protozoa in the rumen was analysed. Partial fragments of archaeal SSU rRNA genes were amplified from DNA isolated from single cells from the rumen protozoal species Metadinium medium, Entodinium furca, Ophryoscolex caudatus and Diplodinium dentatum. Sequence analysis of these fragments indicated that although all of the new isolates clustered with sequences previously described for methanogens, there was a difference in the relative distribution of sequences detected here as compared to that of previous work. In addition, many of the novel sequences, although clearly of archaeal origin have relatively low identity to the sequences in database which are most closely related to them.
AIMS: To investigate the presence of GATC-specific modification and restriction activities in rumen isolates of Treponema sp. METHODS: The presence of N6-methyladenine within GATC (Dam) sequences was analysed using isoschizomeric restriction endonucleases having different sensitivities to the methylation of the target sequence. A fast screening method was used for testing of site-specific endonuclease activities directly in crude cell extracts. Three out of six rumen isolates of Treponema sp. showed restriction activities. Restriction endonucleases were further purified by Heparin-Sepharose chromatography. Using PCR and specific primers, no sequence homologous to the T. pallidum dam gene was found. CONCLUSIONS: Three rumen treponemal strains were documented to possess MboI isoschizomeric restriction-modification systems. SIGNIFICANCE: This is the first report on restriction activity in rumen treponemes.
Two sheep were equipped with rumen and omasal fistulae. A teflon tube with 1 mm outer diameter (OD) was permanently fixed between the abomasal fistula and the reticulo-omasal orifice. The effluent sampling tube was introduced through the abomasal cannula, through the omasal canal into the reticulo-omasal orifice by aid of the teflon tube. The technique allowed direct collection of reticular effluent from sheep. Intermittent flow of reticular effluent was observed. The moments of flow coincided with the end of the second phase of reticular contractions. From the flow pattern it was concluded that the motility of the reticulo-omasal orifice was not affected by the sampling procedure. From the similarities in the pH values, DM contents, and total protozoal counts in the effluent and in the reticular samples, it was concluded that representative samples of effluent could be obtained by the method. One animal was used for sampling for a period of eight months and then again 2 1/2 years later. A fraction of 97 to 98% of the DM present in the reticular effluent and in the reticular samples passed a sieve with 1 mm pore size. From the analyses of reticular content and reticular effluent it was concluded that the reticulo-omasal orifice does not have a significant discriminating function for the selective passage of particles and protozoa into the omasum.
1. Protozoa in rumen contents and omasal effluent of growing wethers were counted. The wethers were equipped with rumen and abomasal cannulas, and omasal sleeves attached to the omasal-abomasal orifice. Rumen fluid dilution rates were elevated by continuous infusions of hypertonic mineral solutions (3-4 litres/d) for 24 d. Rumen contents and omasal effluent were sampled between 9 and 21 h during the last 10 d of each experiment. 2. Protozoal concentrations in omasal effluent were only 0.2-0.3 those found in the rumen under normal conditions. The ratio of protozoal concentrations in rumen: those in omasal effluent was for small Diplodinium spp. 4.6 (SD 0.9), for Ophryoscolex spp. 4.3 (SD 1.0), for Dasytricha ruminantium 4.0 (SD 0.5), for Isotricha spp. 3.8 (SD 0.8), for Entodinium spp. 3.6 (SD 0.9) and for Polyplastron multivesiculatum 2.6 (SD 0.5). 3. Elevation of rumen fluid dilution rate by 20 and 55% respectively, increased protozoal concentrations in omasal effluents from 22 to 33% and from 31 to 47% those in rumen contents. The apparent residence times of protozoa in the rumen were decreased 50% by the infusion of a mineral-salt solution. The increase in rumen fluid dilution rate had no significant effect on concentrations of protozoa in the rumen or on the differences of the apparent residence times between different species. The apparent residence time of holotrichs remained the same before and after infusion of the mineral-salt solution. 4. Apparent residence times of individual species of protozoa in the rumen were, under normal feeding conditions, 2.55 d, and were four to six times longer than the mean residence time of CrEDTA in the rumen.
When buffalo were fed once daily, significant diurnal variations in concentration of rumen ciliates and occurrence of dividing protozoa were found. Differences in proportions of dividing Entodinium- and Diplodinium-type ciliates were also observed. Results obtained suggest that the range of diurnal fluctuations in rumen protozoa concentration may be related to the percentage of dividing cells in populaitons of these organisms.