Marcin Taciak

The effects of inulin, dried Jerusalem artichoke tuber and a multispecies probiotic preparation on microbiota ecology and immune status of the large intestine in young pigs

ABSTRACT The study aimed at determining the effect of two types of prebiotics and a multispecies probiotic on microbiota activity and composition, as well as mucosal immunity in the large intestine of young pigs. In total 48 piglets were divided into 6 groups (n = 8), which received from day 10 of life probiotic-unsupplemented (PU) or probiotic-supplemented (PS) diets. Probiotics were added at 0.5 g/kg diet and contained: Lactococcus lactis, Carnobacterium divergens, Lactobacillus casei, Lactobacillus plantarum and Saccharomyces cerevisiae. The PU and PS diets were formulated without prebiotic addition (control) or with addition of 2% of inulin from chicory root (IN) or 4% of dried Jerusalem artichoke tubers (DJA). After 40 days of feeding, digesta and tissue samples were taken from the caecum and three sections of the colon for analyses of microbiota activity and composition, secretory immunoglobulin A (sIgA) and intraepithelial lymphocytes (IEL). IN diets decreased the caecal digesta pH and β-glucosidase activity but increased propionic, valeric and total short chain fatty acid (SCFA) concentrations compared to control diets. Feeding DJA diets increased caecal valeric acid level, decreased the concentration of isoacids in the colon, reduced β-glucosidase and β-glucuronidase activity in the middle colon and increased Bifidobacterium spp. populations in the proximal and distal colon. PS diets increased the caecal acetic acid and total SCFA level, and Clostridium spp. populations in the distal colon. Neither probiotic nor prebiotics affected sIgA level or IEL number in the large intestine. In conclusion, DJA modified the microbiota ecology in the large intestine of young pigs to a greater extent than IN and the applied probiotic did not enhance effects of prebiotics.

Interactive Effects of Indigestible Carbohydrates, Protein Type, and Protein Level on Biomarkers of Large Intestine Health in Rats.

The effects of indigestible carbohydrates, protein type, and protein level on large intestine health were examined in rats. For 21 days, 12 groups of six 12-week-old male Wistar rats were fed diets with casein (CAS), or potato protein concentrate (PPC), providing 14% (lower protein level; LP), or 20% (higher protein level; HP) protein, and containing cellulose, resistant potato starch, or pectin. Fermentation end-products, pH, and β-glucuronidase levels in cecal digesta, and ammonia levels in colonic digesta were determined. Cecal digesta, tissue weights, cecal and colon morphology, and colonocyte DNA damage were also analyzed. Digesta pH was lower, whereas relative mass of cecal tissue and digesta were higher in rats fed pectin diets than in those fed cellulose. Cecal parameters were greater in rats fed PPC and HP diets than in those fed CAS and LP diets, respectively. Short-chain fatty acid (SCFA) concentrations were unaffected by protein or carbohydrate type. Total SCFA, acetic acid, and propionic acid concentrations were greater in rats fed LP diets than in those fed HP. Cecal pool of isobutyric and isovaleric acids was greater in rats fed PPC than in those fed CAS diets. PPC diets decreased phenol concentration and increased ammonia concentration in cecal and colonic digesta, respectively. Cecal crypt depth was greater in rats fed PPC and HP diets, and was unaffected by carbohydrates; whereas colonic crypt depth was greater in rats fed cellulose. Myenteron thickness in the cecum was unaffected by nutrition, but was greater in the colon of rats fed cellulose. Colonocyte DNA damage was greater in rats fed LP diets than in those fed HP diets, and was unaffected by carbohydrate or protein type. It was found that nutritional factors decreasing cecal digesta weight contribute to greater phenol production, increased DNA damage, and reduced ammonia concentration in the colon.

Quantification of the level of fat-soluble vitamins in feed based on the novel microemulsion electrokinetic chromatography (MEEKC) method

BACKGROUND Simultaneous quantification of liposoluble vitamins is not a new area of interest, since these compounds co-determine the nutritional quality of food and feed, a field widely explored in the human and animal diet. However, the development of appropriate methods is still a matter of concern, especially when the vitamin composition is highly complex, as is the case with feed designated for laboratory animals, representing a higher health and microbiological status. RESULTS A method combining microemulsion electrokinetic chromatography (MEEKC) with liquid-liquid extraction was developed for the determination of four fat-soluble vitamins in animal feed. A separation medium consisting of 25 mmol L⁻¹ phosphate buffer (pH 2.5), 2-propanol, 1-butanol, sodium dodecyl sulfate and octane allowed the simultaneous determination of vitamins A, D, E and K within a reasonable time of 25 min. The polarity of the separation voltage was reversed in view of the strongly suppressed electro-osmotic flow, and the applied voltage was set at 12 kV. The fat-soluble vitamins were separated in the order of decreasing hydrophobicity. CONCLUSION It was proved that the proposed MEEKC method was sufficiently specific and sensitive for screening fat-soluble vitamins in animal feed samples after their sterilization.

Nutritional value and physiological effects of soya-free diets fed to rats during growth and reproduction.

The objective of the study was to assess the effects of substitution milk and egg for soya products in breeding diets for rats, with concomitant decrease of the dietary protein level and supplementation with amino acids. Soya-containing (S) and two soya-free (NS and NSA) diets were evaluated as protein and energy sources, and their effects on reproductive performance during two cycles, and on the quality of the offspring were assessed. Organ weights were registered in females and blood parameters were determined in males. In the offspring males from S and NS groups, plasma LH, testosterone and prolactin levels were measured on the 22nd and the 60th day of life. The S diet contained more protein of smaller concentration of methionine and cystine and lower biological value than both NS and NSA diets and promoted similar post-weaning growth rate, similar body weight changes of dams during gestation and lactation and slightly lower mating efficiency. Within each reproductive cycle, the number and individual and total body weight of newborn and weanling pups did not differ but in two cycles mean number of neonates per litter and mean litter weight were significantly lower on S than on NSA diet. Plasma concentration of hormones did not differ in 22-day-old offspring males while in the older ones LH and prolactin levels were higher in animals fed on S than on NS diet. It is concluded that replacing soya protein by milk and egg protein with concomitant lowering dietary protein level and amino acid supplementation does not impair the growth rate and tends to improve reproductive performance. Feeding soya-free vs. soya-containing diets differentiates hormonal status of young males.

Health relevance of intestinal protein fermentation in young pigs.

Abstract The physiological role of the gastrointestinal microbiota has become an important subject of nutrition research in pigs in the past years, and the importance of intestinal microbial activity in the etiology of disease is doubtless. This review summarizes the recent knowledge related to the microbial ecology of protein fermentation and the appearance of protein-derived metabolites along the pig intestine. The amount of fermentable protein depends on factors such as dietary protein concentration, protein digestibility due to secondary or tertiary structure, the interaction with dietary compounds or anti-nutritional factors, and the secretion of endogenous proteins into the gut lumen. High protein diets increase the luminal concentrations and epithelial exposure to putatively toxic metabolites and increase the risk for post-weaning diarrhea, but the mechanisms are not yet clarified. Although the use of fermentable carbohydrates to reduce harmful protein-derived metabolites in pigs is well-established, recent studies suggest that the inclusion of fermentable carbohydrates into diets with low protein digestibility or high dietary protein level may not ameliorate all negative effects with regard to epithelial response. Based on the current knowledge, the use of diets with low levels of high-quality protein may help to reduce the risk for intestinal disease in young pigs.

Physiological effects of long-term feeding diets supplemented with potato fibre or cellulose to adult rats

Nutritional value of diets containing 10% potato fibre or cellulose (diets MPF and MC, respectively) and their long-term effects on parameters related to potential health benefits were studied in rats. In a first experiment nutrient digestibility and metabolisable energy of the diets were determined. In a second experiment, each diet was fed to 20 male and 20 female rats aged nine weeks old during 12 (Lot W12) or 24 (Lot W24) weeks. Feed intakes per cage and body weight of rats were registered. In all animals organ weight was determined and in animals of Lot W12 and in males of Lot W24 biochemical blood parameters were analysed. In males of Lot W24, caecal SCFA concentration and body composition were also determined. Testosterone blood concentration was determined in males of Lots W12 and W24. Protein and fat digestibilities were lower, while those of ash, crude fibre and NFE were higher in the MPF than the MC diet. The energy value of the MPF diet was greater but energy intake was similar for both diets. Body weight, body protein and fat contents in males of Lot W24 did not differ between the groups, while body ash was significantly greater in MPF than MC rats (2.64 vs. 2.17 g/100 g BW). SCFA concentration and pool were considerably greater in MPF than MC rats, the SCFA profile being similar. Potato fibre did not affect total cholesterol, whereas it decreased triglycerides and very low density lipoproteins (VLDL) in Lot W12 of both genders, and tended to decrease them in males of Lot W24. Blood glucose and testosterone concentrations and relative mass of prostate were significantly depressed by the MPF diet in males of Lot W24. It can be concluded that feeding potato fibre stimulates caecal fermentation, improves mineral absorption and does not affect total cholesterol levels. The concentration of lipids, VLDL, glucose and testosterone depended on potato fibre in an age-dependent way.

Influence of different inclusion levels and chain length of inulin on microbial ecology and the state of mucosal protective barrier in the large intestine of young pigs

The objective of the study was to examine the effect of inulin level, in regard to its degree of polymerisation (DP), on microbial activity and mucosal immune system of the large intestine of pigs. A total of 56 castrated male piglets (PIC × Penarlan P76) were allocated to seven groups and fed from the 10th day of life cereal-based diets without the addition of inulin or with 1%, 2% or 3% of inulin with an average DP of 10 (IN10) or 23 (IN23). Pigs were sacrificed at the age of 50 days. Feeding IN10 diets increased fructan concentration in the large intestine compared with IN23 diets, but did not affect microbial activity, except for digesta pH and mucinase activity in the middle colon, which were greater at the 1% level compared with the control group and other IN10 diets, respectively. The concentration of secretory immunoglobulin A in the caecum and middle colon was reduced by the 1% IN10 diet compared with the control group. Pigs fed the 2% IN23 diet had a higher butyric acid concentration in the caecum and proximal colon and greater isoacid concentrations in the middle and distal colon in comparison to the control. Dietary level of IN23 did not affect secretory immunoglobulin A concentration but the count of caecal intraepithelial lymphocytes was higher in pigs on the 1% IN23 diet than on the control diet. Neither IN10 nor IN23 diets affected populations of Bifidobacterium or Lactobacillus spp. In conclusion, the effects of inulin in the large intestine of pigs depended on dietary level and DP. IN23 increased short-chain fatty acid production at the 2% level and slightly activated mucosal immune status at the 1% level.

Determination of Water Soluble Vitamins in Laboratory Animal Feeds by Micellar Electrokinetic Chromatography

Seven water-soluble vitamins: thiamine hydrochloride (B1), riboflavin hydrochloride (B2), pyridoxine hydrochloride (B6), calcium pantotenate (B5), ascorbic acid (C), folic acid (B9), and biotin (H, B7) were separated using the micellar electrokinetic chromatographic (MEKC) method in a single run with the electrolyte solution consisting of 100 mM H3BO3, 5 mM Na2B4O7, and 30 mM sodium dodecylsulphate (SDS). The separation was achieved within 8 min. and the detection was performed at 200 nm. The calibration graphs plotted with six concentrations of each vitamin were linear with the determination coefficient r > 0.999. The method was applied for quantification of vitamins B1, B2, B5, B6, B9, C, and H in novel feed for laboratory animals. The sample preparation involved solid-phase extraction of the vitamins, used prior to the MEKC analysis. The attained precision level was good with the recoveries between 98.4% and 105.6%. The simplicity of the procedure should make it highly useful for feed quality control in husbandry and the animal feed industry.

The effects of feeding diets containing potato protein concentrate on reproductive performance of rats and quality of the offspring.

Two diets containing either soybean meal (SBM) or SBM and potato protein concentrate (PPC) in proportion of protein 1:1 (diet BS and BPP, respectively), were fed each to 22 females and 22 males from the age of 4-5 weeks throughout two reproductive cycles. A commercial closed-formula SNF diet containing unknown amount of potato protein was fed till the end of the first reproductive cycle. All three diets were also fed during 21 days following weaning, each to 10 offspring males born in the first cycle. The solanidine glycoalkaloids (GA) content in PPC was 2316 mg/kg DM. Feeding diets containing potato protein did not affect pregnancy rate, number and conformation of neonates and number of pups weaned (in all litters number of neonates left with dams being reduced to eight) but it depressed their body weight (weight of weaners on BPP diet only in the first cycle). Growth rate of the offspring males did not differ but feed intake was lower and feed conversion was higher on BPP than on BS and SNF diets. Feeding of BPP diet to offspring did not affect mass of body organs but increased blood urea and albumin concentrations while feeding SNF diet increased relative weight of small intestine and liver and raised blood concentrations of alanine amino transferase, albumin, protein, urea and cholesterol. In dams sacrificed after two cycles only relative weight of uterus and ovaries were lower in females fed on BPP than on BS diet. It is concluded that feeding moderate amounts (6.4%) of high-GA PPC does not impair reproductive performance, does not induce malformation of pups but depresses foetal and postnatal growth rate till weaning but not after weaning.

The fructanolytic abilities of the rumen bacterium Butyrivibrio fibrisolvens strain 3071

To determine if Butyrivibrio fibrisolvens strain 3071 is able to use fructose polymers for growth and to identify the enzymes involved in their digestion.