B. Pastuszewska

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.

Profile of fatty acids, muscle structure and shear force of musculus longissimus dorsi (MLD) in growing pigs as affected by energy and protein or protein restriction followed by realimentation

Forty-eight gilts were submitted to a 30% restriction of feed (groups F and F1) or protein intake (group P) from 90 to 118 days of age, followed by realimentation from 119 to 168 days of age. Control pigs (C) were fed during the whole experiment according to a semi ad libitum scale. During realimentation all animals were fed according to semi ad libitum scale except pigs F1 which were fed ad libitum. Six pigs from each group were slaughtered at the end of restriction and realimentation. Restriction decreased the weight of musculus longissimus dorsi (MLD) and increased shear force. Restriction of feed intake depressed MUFA concentration and increased n-6/n-3 ratio while protein restriction decreased n-6/n-3 and PUFA:SFA ratios. Structure of fibers was not affected. After realimentation MLD mass was still lower in all previously restricted pigs, shear force was the lowest in F1 pigs. Only percentage of fast twich oxidative fibers was significantly greater in F1 pigs than in others. Significant correlations between parameters investigated during the study were found.

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.

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.

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.

Growth performance and physiological parameters of conventional and specified pathogen‐free rats fed autoclaved diets with different protein sources

The effects of feeding autoclaved commercial SSNIFF (SN) diet and diets containing soya bean (S) and casein (C) to growing conventional (CON) and specified pathogen-free (SPF) rats were determined. Diets S, C and SN, autoclaved at 121 °C during 20 min (T1), at 134 °C during 10 min (T2) and non-autoclaved (T0), were fed during four weeks, each to 8 CON males and 8 females of mean initial body weight 56 g, kept individually. Diets S, C and SN, autoclaved at T1, were fed during two months, each to 20 SPF males and 20 females of mean initial body weight 58 g, kept in group of 5 animals per cage. In CON rats, autoclaving did not affect feed intake and weight gain, decreased thyroid and stomach weight, increased caecal tissue and digesta weight, and concentrations of isobutyric, isovaleric and valeric acid in caecal digesta. Among biochemical blood parameters, autoclaving decreased only total protein concentration and aspartate aminotransferase activity. Feeding C diet resulted in lower feed intake and weight gain in CON and SPF males. Diet affected organ weights and the greatest differences were found in rats on SN diet for weights of stomach, caecum and female reproductive organs. Diet affected concentration of all short-chain fatty acids, pH and weight of caecal digesta, the most important being the greatest butyric acid concentration on SN diet and isoacids on C diet. It is concluded that autoclaving of both soya-containing and soya-free diets does not affect negatively animal performance and physiology.

Effects of dietary fat on the saturated and monounsaturated fatty acid metabolism in growing pigs

ABSTRACT The effect of dietary fats differing in fatty acid (FA) composition on the metabolism of saturated FA (SFA) and monounsaturated FA (MUFA) in growing pigs was investigated. The deposition of FA in the body and the fate of individual dietary FA were assessed after slaughter. Gilts with an initial body weight (BW) of 60 kg were used as experimental animals. Six pigs were slaughtered at 60 kg BW, while further 18 pigs received three isoenergetic and isonitrogen experimental diets containing linseed oil, rapeseed oil or beef tallow at 50 g/kg diet until they reached 105 kg (six pigs per group). The chemical composition and the content of FA in the whole body were determined and compared across groups. Regardless of dietary treatment, the whole body contained similar amounts of protein, fat and total FA. The total accumulation (percentage of net intake and de novo production) of SFA and MUFA was similar in all groups, but the processes of elongation and desaturation of SFA and MUFA depended upon the type of FA added to the diet. A high dietary content and intake of MUFA inhibits desaturation compared to SFA- and PUFA-rich diets, whereas a high SFA content and intake lowers elongation rate. The increasing net intake of total SFA and MUFA was associated with a lower total de novo production of these FA in the whole body of pigs.