The influence of omega-3 polyunsaturated fatty acids feeding on composition of fatty acids in fatty tissues and eggs of laying hens.

The paper deals with supplementation of hen diet with oils with increased content of omega-3 polyunsaturated fatty acids (PUFA) and occurrence of fatty acids (FA) in fatty tissues and eggs of laying hens after the respective supplementation. The experiment was carried out on 30 laying hens of ISA BROWN hybrid, divided to three groups (A, B, C). For the period of three weeks the group A was administered flax-seed oil, group B served as a control and group C was fed base rations supplemented with fish oil. In addition to that the diet of groups A and C was supplemented with vitamin E as an antioxidant. Administration of the oils as a source of omega-3 PUFA increased their concentration in both the fatty tissues and egg-yolk which resulted in the change of the ratio of omega-6 and omega-3 PUFA. No influence of flax-seed and fish oil on concentration of alpha-tocopherol in egg-yolk was observed.

Influencing the immune parameters in germ-free piglets by administration of seal oil with increased content of omega-3 PUFA.

Oral administration of oil with an increased content of omega-3 polyunsaturated fatty acids to germ-free piglets resulted in a significant increase in the total values of CD4, CD8 lymphocytes, B lymphocytes, and monocytes, in peripheral blood in comparison with the controls. The metabolic activities of phagocytes as well as the polyclonal activation of lymphocytes were not significantly influenced. The level of growth factor was significantly higher, as determined on the basis of somatomedin in the blood serum. Biochemical indices showed a significant increase in the level of eicosapentaenoic and docosahexaenoic acids in blood serum and the decrease in the level of arachidonic acid at the same time.

Effect of oil containing n-3 polyunsaturated fatty acids (PUFA) on the immune response and growth factors in piglets.

Oral administration of n-3 polyunsaturated fatty acids (PUFA) to piglets slightly enhances the immune response. As compared to the control, in the experimental piglets the absolute values of monocytes in the peripheral blood were significantly increased (P < 0.05), while the metabolic activity of phagocytes and the number of lymphocytes within the individual subpopulations were slightly higher. The level of growth factors, determined on the basis of somatomedin in the blood serum, was significantly higher in the experimental group (P < 0.05). n-3 PUFA interfere with the synthesis of prostaglandins and influence the metabolism of fatty acids. This finding may contribute to the therapy of inflammatory processes influencing immune and growth factors in piglets.

Potentiation of the effectiveness of Lactobacillus casei in the prevention of E. coli induced diarrhea in conventional and gnotobiotic pigs.

The influence of preventive administration of Lactobacillus casei subsp. casei and maltodextrin KMS X-70 on Escherichia coli 08: K88 adhesion in the gastrointestinal tract of 11 conventional and 6 gnotobiotic piglets was investigated. The preventive administration of L. casei alone had almost no inhibitory effect on the adherence of E. coli to the jejunal mucosa of gnotobiotic and conventional piglets while the lactobacilli administered together with maltodextrin decreased the number of E. coli colonising jejunal mucosa of gnotobiotic piglets by 1 logarithm (4.95 log 10/cm2) in comparison with the control group (5.96 log 10/cm2). L. casei administered in combination with maltodextrin decreased the number of E. coli colonising the jejunum of conventional piglets by more than two and half logarithm (4.75 log 10/cm2, p < 0.05) in comparison with the control (7.42 log 10/cm2). The inhibitory effect of Lactobacillus casei and maltodextrin KMS X-70 on the adhesion of E. coli to the intestinal mucosa of conventional and gnotobiotic pigs was probably mediated by Lactobacillus--produced antibacterial substances and stimulation of immunity.

The effect of lactic acid bacteria on intestinal metabolism and metabolic profile in gnotobiotic pigs.

The effect of inoculation of Lactobacillus casei on selected parameters of metabolic profile and intestinal metabolism of gnotobiotic piglets was investigated during the first three weeks of their life. The experiment was carried out on 8 germ-free piglets. The experimental group was inoculated once a day with the Lactobacillus casei subsp. casei strain. The inoculum contained 1 x 10(8) microorganisms in 1 ml. The control group of piglets received no inoculum. Lactobacillus casei colonized jejunum and ileum in the numbers from 5.63 to 6.06 log 10 cm-2 and their numbers in the jejunal and ileal contents were in the range 8.38-9.87 log 10.ml-1. The daily consumption of milk by the inoculated animals was significantly higher (p < 0.001). The average weight of inoculated piglets at the end of the period investigated was higher by 29.7%. Lactobacillus casei affected several parameters investigated. Piglets inoculated with lactobacilli showed significantly lower (p < 0.05-0.01) values of pH of the jejunal content, numbers of erythrocytes, values of haematocrit, urea, glucose, total lipids, cholesterol and calcium in the serum and significantly higher values (p < 0.05-0.01) of lactic acid in the jejunal content. The values of phagocytic activity and the index of phagocytic activity in the piglets of the experimental group were two to three-fold higher in comparison with those detected in the control group. The application of Lactobacillus casei affected positively the growth of gnotobiotic piglets, their intestinal metabolism, the level of cholesterol in the serum and phagocytic activity.

In vitro studies of porcine lactobacilli for possible probiotic use.

Fourteen strains of lactobacilli isolated from the gut of suckling pigs were identified and studied to determine their susceptibility to antimicrobial feed additives, acid tolerance, adherence to epithelial cells from the porcine intestine and antimicrobial activity. Four strains were identified as Lactobacillus casei subsp. casei, two strains as Lactobacillus rhamnosus as well as Lactobacillus reuteri and three strains as Lactobacillus salivarius. The remaining strains could not be identified. Antimicrobial susceptibility tests showed susceptibility of the lactobacilli to many of the antimicrobial feed additives commonly used in pigs. Strains resistant to cyadox and nourseothricin occurred in high numbers. The tested lactobacilli were acid-resistant at pH 3. Thirteen strains of lactobacilli exhibited strong adherence to epithelial cells from the porcine intestine while one strain was classed as weakly-adherent. All lactobacilli showed inhibitory activity against indicator bacteria in the presence of glucose. Lactate and acetate were the principal final products of glucose fermentation in all strains. Only three strains produced H2O2 in detectable amounts.

The effect of lactobacilli inoculation on organic acid levels in the mucosal film and the small intestine contents in gnotobiotic pigs.

The effect of the inoculation of three Lactobacillus plantarum strains upon lactic, acetic, acetoacetic and propionic acid levels in the mucosal film (F) and the jejunal and ileal contents (O) has been investigated in gnotobiotic pigs. In the jejunum of the inoculated animals, the mucosal film revealed significantly increased levels of lactic, propionic and acetoacetic acids when compared to the contents (25.3 vs. 10.8 mmol.l-1, 18.5 vs. 5 mmol.l-1 and 29.7 vs. 11.2 mmol.l-1, respectively) as well as insignificantly increased acetic acid levels (11.0 vs. 5.8 mmol.l-1). In the ileum of gnotobiotic pigs, propionic acid levels of the mucosal film were significantly higher than those of the contents (21.2 vs. 9.5 mmol.l-1, p < 0.05). In comparison to the contents, the increased lactic, acetic and acetoacetic acid levels in the film proved to be insignificant. The above results suggest that the significantly increased levels of the Lactobacilli-produced organic acids may present an efficient barrier inhibiting the adherence of digestive tract pathogens to the intestinal mucosa.

Interactions of Lactobacillus spp. and enteropathogenic Escherichia coli under in vitro and in vivo conditions.

In the present study, the effect of Lactobacillus spp. against enteropathogenic Escherichia coli 08:K88+Ent+ under in vitro and in vivo conditions has been compared and the effect of inoculation of Lactobacillus spp. upon the colonization of both the jejunum and ileum by enteropathogenic E. coli 08:K88+Ent+ in 9 gnotobiotic pigs has been observed. Under in vitro conditions, the strain Lactobacillus spp. showed the inhibition of 2.1 +/- 0.1 mm against enteropathogenic E. coli 08:K88+Ent+. Two days after the inoculation, the enteropathogenic E. coli 08:K88+Ent+, inoculated to the control group of gnotobiotic pigs (E), colonized the mucosa of both jejunum and ileum of gnotobiotic pigs at counts of 6.41 and 6.08 log 10/cm2, respectively. In experimental group (L-E), the counts of adhered enteropathogenic E. coli in the identical sections of the small intestine, following the inoculation by Lactobacillus spp., amounted to 6.35 and 6.43 log/cm2, respectively. In both groups, numbers of E. coli in the intestinal content of both jejunum and ileum were nearly the same (group E 9.03 and 9.31 log 10/ml; group L-E 8.97 and 9.11 log 10/ml). Two to five days after E. coli inoculation, Lactobacillus spp. counts adhered to the jejunal wall ranged from 5.4 to 6.49 log 10 cm2; in the ileum they ranged from 6.05 to 6.77 log 10 cm2. In the jejunal content, the lactobacilli counts ranged from 6.81 to 8.86 log 10/ml and in the ileum from 8.5 to 898 log 10/ml. Two days after the E. coli inoculation, the concentration of lactic acid in the content of jejunum in gnotobiotic pigs of the groups E and L-E was 16.3 mmol/l and 23.6 mmol/l, respectively. The concentration of acetic acid in the jejunum of the pigs of E and L-E groups was 15.9 mmol/l and 19.6 mmol/l, respectively. Similarly, the higher concentrations of both acids were found also in the ileum of the L-E pigs. The results obtained indicate that the used strain of Lactobacillus spp. which has been preventively inoculated to gnotobiotic pigs, did not prevent the adhesion of enteropathogenic E. coli 08:K88+Ent+ to the mucosa of both jejunum and ileum also despite of the demonstrated inhibitory effect against enteropathogenic E. coli under in vitro conditions and despite good adherent ability in vivo. In both groups of animals, the diseases with pronounced clinical signs as well as losses have occurred.

Indirect examination of exocrine pancreatic function in vivo by determination of chymotrypsin activity in animal intestine using a synthetic substrate.

Using the substrate N-acetyl-L-tyrosyl-p-aminobenzoic acid, we determined chymotrypsin activity in the small intestine of calf, pig, and poultry. Orally administered N-acetyl-L-tyrosyl-p-aminobenzoic acid is enzymatically cleaved in vivo, and the released p-aminobenzoic acid is determined by HPLC. We found that the p-aminobenzoic acid concentration in plasma and urine was significantly influenced by the feeding of soya flour. After soybean flour feeding, the p-aminobenzoic acid concentration significantly increased in the plasma of calves and hens, in contrast to pigs, where the p-aminobenzoic acid concentration significantly decreased. This shows that the oral administration of N-acetyl-L-tyrosyl-p-aminobenzoic acid with subsequent determination of p-aminobenzoic acid is suitable for the estimation of exocrine pancreatic function and for determination of changes in intestinal proteolytic activity caused by antinutritive substances.

Simultaneous determination of p-aminobenzoic acid and its metabolites in urine by high performance liquid chromatography.

We developed a new HPLC method for the determination of p-aminobenzoic acid (PABA) and its metabolites (p-aminohippuric acid, N-acetyl-p-aminohippuric acid, N-acetyl-p-aminobenzoic acid) in urine. As the internal standard m-hydroxybenzoic acid was used. In the isocratic elution the mobile phase consisted of methanol and 0.02 M ammonium acetate (20:80 v/v, pH 4.0). The separation was carried out on the C18, reversed-phase column, particle size 5 microns. The separated components were detected at 280 nm. The method can be used in the assessment of the response of pancrease (secretion of digestive enzymes) to soya feeding as well as in the diagnosis of the exocrine pancreatic diseases of animals.

[Colonization of the digestive tract by defined lactoflora in gnotobiotic and conventional lambs]. / Kolonizácia tráviaceho traktu gnotobiotických a konvencných jahniat definovanou laktoflórou.

The effect of Lactobacillus casei 249/89 on the colonization of the intestinal tract and selected indicators of the metabolic profile of gnotobiotic and conventional lambs from the viewpoint of its possible utilization in the prevention of diarrhoeal syndrome of bacterial etiology in young animals has been studied. Five gnotobiotic and three conventional lambs were used in these studies. The lambs were slaughtered at 3, 6, 10, 15 and 21 days of age. The population of L. casei colonizing the intestinal epithelium was at the age dynamics on average higher in gnotobiotic lambs compared with lactobacilli in conventional lambs (Fig. 1). The significant difference (p < 0.001) was noted at the age of 3 days (gnotobiotic lambs = 3.40 log 10/cm2 and conventional lambs = 1.08 log 10/cm2). Also the counts of lactobacilli colonizing individual sections of the intestine (Fig. 2) was on average higher in gnotobiots with significant differences in jejunum and colon (p < 0.05). In both groups, the highest number of lactobacilli was observed on the intestinal epithelium in lower sections of the digestive tract. In gnotobiots, the population of L. casei colonizing the duodenum was highest at the age of 3 days (3.49 log 10/cm2), in jejunum (3.74 log 10/cm2) and in ileum 4.37 log 10/cm2) at the age of 6 days and in the colon (4.7 log 10/cm2) at the age of 15 days (Fig. 3). In conventional lambs, the population of lactobacilli colonizing individual sections of the intestinum was increasing with age (Fig. 4). The number of L. casei in the intestinal content of gnotobiotic lambs was on average higher than that in conventional animals but the differences were not significant. In both groups, the highest number of lactobacilli was noted at the age of 10 days (gnotobiotic lambs = 5.9 log 10/ml; the conventional lambs = 4.6 log 10/ml). From the viewpoint of individual sections of the intestine, the population of L. casei in the intestinal content of gnotobiotic lambs was higher than the lactobacilli count in conventional animals with significant difference in jejunum (p < 0.01). In gnotobiots, the highest lactobacilli count was in the colon (6.17 log 10/ml); in conventional lambs in the ileum (4.71 log 10/ml).(ABSTRACT TRUNCATED AT 400 WORDS)