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V Kmet

Publications and source records attributed to V Kmet.

32 records · Page 2Linked to original sources

Isolation and partial characterization of three rumen Lactobacillus plantarum bacteriophages.

The first isolation of Lactobacillus plantarum bacteriophages from ruminal fluid is reported. Three bacteriophages were characterized on the basis of plaque morphology, host ranges, stability, electron microscopic morphology and DNA restriction endonuclease digestion patterns. They formed clear plaques and are placed in group A of Bradley's scheme and have identical host ranges. Bacteriophages were stable to urea and chloroform. They were relatively thermostable but partially inactivated by rumen fluid and by acetate. DNA restriction analysis showed that phage L20 had different numbers of cleavage sites in comparison with the next two phages.

Acetates↗

Adherence of ruminal Streptococcus bovis and Lactobacillus strains to primary and secondary cultures of rumen epithelium.

Six strains of rumen Lactobacillus and four Streptococcus bovis strains isolated from rumen wall and fluid samples were examined for the adherence to cells of primary and secondary cultures of ruminal epithelium (REC) prepared from sheep and calf. S. bovis adhered to the keratinized REC. Ruminal lactobacilli did not adhere. The presence of rumen lactobacilli in mixture had no influence on the adherence of S. bovis strains. No difference was observed in the adherence of tested bacteria to epithelial cells of primary or secondary cultures, but adhesion was only detected on keratinized cells.

Animals↗

Preliminary observations of interaction between bacteriophages and Streptococcus bovis bacteria on ruminal epithelium primoculture.

Five Streptococcus bovis strains (47/3, 59/2, 4/1, 46/2 and 44/9) isolated from calf ruminal fluid samples were examined for the adherence to cultured ruminal epithelium cells. Four strains (47/3, 59/2, 4/1 and 46/2) were able to attach to the cultured epithelial cells. However, S. bovis 47/3 strain attached to the target cells in significantly greater numbers than the other strains. Strain 44/9 did not adhere to cells of ruminal epithelium. The adherent bacteria were observed on the surface of differentiated (mainly keratinized) cells of ruminal epithelium primoculture only. The different effect of F4, F5 and F6 bacteriophages was ascertained on S. bovis bacteria adhering to rumen epithelial primoculture. A significant decrease in the number of adherent bacteria was shown after cultivation of strains 47/3 and 4/1 with F6 bacteriophage and of 47/3 strain with F4 phage. The F5 bacteriophage had no significant effect on these bacteria.

Animals↗

Occurrence of conjugative amylolytic activity in rumen lactobacilli.

1,263 rumen strains of Lactobacilli were selected for the production of amylase and resistance to antibiotics. Out of 48 amylolytic Lactobacilli, in 17 strains, there was a cotransfer of the alpha amylase activity and resistance to tetracyclin with a transfer frequency of 10(-3) to 10(-7) ml-1 of the conjugative mixture. In strains No. 30, No. 36 and No. 49, plasmids with similar molecular weights as plasmids in corresponding transconjugants were detected. Similar protein pattern of the donors recipient of the L. lactis RN 1 and transconjugants obtained by PAGE proved the cotransfer of the amylase activity and antibiotic resistance.

Amylases↗

Isolation and characterization of two rumen Streptococcus bovis bacteriophages.

A method for the isolation of Streptococcus bovis bacteriophages from ruminal fluid of calves is described. Thirty to 2 x 10(3) phages per ml infecting Streptococcus bovis strains 4/1 and 47/3 were isolated directly from ruminal fluid. Two bacteriophages were characterized on the basis of plaque morphology, host ranges, electron microscopic morphology and DNA restriction endonuclease digestion patterns. The F1 and F3 phages formed clear plaques of different sizes. The plaque size of the F1 phage was about 1-1.5 mm in diameter, while the plaques of the F3 phage were larger (1.5-2.5 mm in diameter). Both phages are placed in group B of Bradley's scheme and have different host ranges. The first isolation of Streptococcus bovis phage DNA is reported. Restriction analysis of their DNAs showed that phages F1 and F3 had different numbers of cleavage sites in their genomes and that they were not identical.

Animals↗

[Production of volatile fatty acids in the rumen of calves during dietary-microbial stimulation and early weaning].

The contents of volatile fatty acids in the rumen fluid were studied in calves at an early (seven weeks of age) and traditional weaning term (nine weeks of age). Weaning at the age of seven weeks enabled to save 16.2 kg of milk replacer per calf; there was also a lower content of concentrates in the starter feed with a supplement of Amylastim. The health condition of the calves was good in both groups. The average daily weight gains for the period from the second to the ninth week of age were 0.550 kg in the early weaned calves and 0.690 kg in the calves weaned at a normal time. Early weaning had a positive influence on the development of rumen metabolism. Calves weaned at the age of seven weeks, compared with those weaned at the age of nine weeks, had much higher concentrations of VFA (p less than 0.05) in rumen fluid (at the age of seven weeks: 130.49 mmol per litre vs. 111.53 mmol per litre; at the age of eight weeks: 119.74 mmol per litre vs. 96.98 mmol per litre). Early weaned calves had the statistically significantly (p less than 0.05) higher proportions of propionic acid, butyric acid, i-butyric acid, and valeric acid. Later-weaned calves had the significantly higher (p less than 0.05) contents of acetic acid and i-valeric acid.

Animal Feed↗

Rumen microbial changes in calves fed on alpha amylase diet.

Fifty-six calves in the period of a milk nutrition, randomly assigned to two groups (control and alpha amylase fed) were used to monitor the changes in the rumen microbial populations and total volatile fatty acids (VFA) concentrations associated with feeding Amylosubtilin G10X (0.6 g.day-1). Statistically significant increase was observed for amylolytic counts in experimental calves on 20th or 30th days. The counts of rumen cellulolytic bacteria and selenomonas tended to have increase in the value, but were not significantly different from those in the control valves. The results show that alpha amylase fed calves has a relative higher rumen microbial population.

Animals↗

Metabolism and some characteristics of lactobacilli isolated from the rumen of young calves.

Fourteen strains of lactobacilli isolated from the rumen of young calves were studied to determine their biochemical characteristics, growth parameters, metabolism on lactose and sensitivity to 28 antimicrobial agents. Thirteen homofermentative strains belonged to Lactobacillus acidophilus and one heterofermentative strain resembled Lact. fermentum. The relevance of rumen lactobacilli to the nutrition of calves is discussed.

Animal Nutritional Physiological Phenomena↗

[The occurrence and properties of Selenomonas ruminantium bacteria in calves during the time of milk feedings].

The occurrence, morphological properties, urease and alpha-amylase activities of Selenomonas ruminantium were investigated in calves in the period of milk nutrition. The average number of the organisms was found to range between 10(7) and 10(8) per 1 millilitre of rumen contents. The alpha-amylase activity of the Selenomonas strains ranged from 0.1 to 8.8 ncat.ml-1 whereas their urease activity ranged from 6.3 to 261 ncat.ml-1 of nutrient medium. The presence of morphologically typical Selenomonas ruminantium and spontaneous formation of spheroplasts were found by electron microscopic examination.

Age Factors↗

Urease activity of adherent bacteria and rumen fluid bacteria.

In experiments on six sheep fed on a low nitrogen diet (3.7 g N/day), urease (EC 3.5.1.5) activity (nkat X mg-1 bacterial dry weight) 3 h after feeding was found to be highest in the bacteria adhering to the rumen wall (13.25 +/- 2.10), lower in the rumen fluid bacteria (8.96 +/- 1.35) and lowest in the bacteria adhering to feed particles in the rumen (5.69 +/- 2.13). The urease activity of bacteria adhering to the rumen wall and of the rumen fluid bacteria of six sheep fed on a high nitrogen diet (21 g N/day) was significantly lower than in sheep with a low N intake and in both cases was roughly the same (3.81 +/- 1.37 and 3.76 +/- 1.02 respectively); it was lowest in bacteria adhering to feed particles in the rumen (1.92 +/- 0.90). It is concluded from the results that the urease activity of rumen fluid bacteria and of bacteria adhering to the rumen wall and to feed particles in the rumen is different and that it falls significantly in the presence of a high nitrogen intake. From the relatively high ureolytic activity of bacteria adhering to the rumen wall in the presence of a low nitrogen intake it is assumed that this is one of the partial mechanisms of the hydrolysis of blood urea entering the rumen across the rumen wall and of its reutilization in the rumen-liver nitrogen cycle in ruminants.

Animal Feed↗

[Use of the colonizing preparation Amylastin in the stimulation of rumen-type digestion in young ruminants].

In experiments with calves during the milk-nutrition period we tested the use of the colonisation preparation Amylastim, containing a bacterial strain of Streptococcus bovis AO 24/85, as a factor speeding up the development of the rumen microflora. We gave the preparation to the animals for a period of four weeks and studied its influence on the microflora adherent to the epithelium of the rumen wall. We ascertained a significant increase in the number of Streptococcus bovis germs (P less than 0.001) as well as of alpha-amylase activity (P less than 0.05). When giving the preparation Amylastim to calves in large-scale production conditions we ascertained a positive effect on the health condition, total losses being lower.

Animals↗

Dynamics of enzymatic activity development in the adherent rumen microflora.

In lambs in the period of a milk nutrition, the enzymatic activities of the rumen wall microflora: cellulase (EC 3.2.1.4), beta-glucanase (EC 3.2.1.73), alpha-amylase (EC 3.2.1.1), urease (EC 3.5.1.3) and neutral protease have been investigated. For comparison's sake, the enzymatic of adherent microflora of the caecal wall ahs been observed, too. It was found that the functionally latent rumen of lambs contained adherent bacteria with cellulase and beta-glucanase activities which reached their maximum on the 30th day of life. The alpha-amylase and protease activities of the epimural rumen microflora reached their maximum already on the 20th day of life. Contrarily, the urease activity reached its maximum only on the 40th day of life. The results show that already from the 10th day after birth, the rumen of young ruminants has a relatively high microbial enzymatic activity.

Animals↗

Effect of pectinase on rumen fermentation in sheep and lambs.

In trial with adult wethers and weaned lambs the effect of enzymatic preparation Pektofoetidin G3x (mostly pectinase and cellulase) on rumen fermentation was studied. After 4 weeks of Pektofoetidin G3x application (0.54 g per day and animal) to adult wethers no statistically significant differences in total volatile fatty acids (VFA), acetic acid, propionic acid, butyric acid, ammonia in the rumen contents and urea in blood were determined between control and enzyme treated group. In comparison of fermentation parameters in wethers (mean of 1-3 hours after feeding) and lambs (2-3 hours after feeding) the significant differences in mol % of acetic acid (63.3 in control, 54.6 in experimental group, P less than 0.01), propionic acid (24.6, vs. 31.3, P less than 0.001), acetate: proprionate ratio (2.54, vs. 1.77, P less than 0.01) and in energy efficiency of VFA production (76.0%, vs. 79.1%, P less than 0.001) were determined. These differences between wethers and lambs suggest more intensive fermentation in lambs than in adult sheep. On the basis of these results it is possible to suggest, that in adult animals the efficiency of application of enzymatic preparations is low. In utilization of enzymatic preparations more important role, probably, is that of ruminal ecosystem itself, that, if once fully developed, is perfectly resistant to biotechnological interferences.

Acetates↗