Evaluation of Acute Phase Proteins in Clinically Healthy Dairy Cows in Perinatal Period and During Lactation.

The estimation of acute phase proteins (APP), which are recognized as inflammation markers is a good method for animal health monitoring. Several factors such as obesity, age and sex are also known to modulate APP status. We evaluated the influence of pregnancy and lactation in 65 clinically healthy dairy Holstein-Friesian dairy cows, 2nd÷4th lactation, chosen from 3 different dairy farms located in South West part of Poland. Bovine C-reactive protein (CRP), haptoglobin and fibrinogen were assayed using commercial ELISA kits. The highest values of CRP and haptoglobin were observed in cows during the first month after calving. The highest concentrations of fibrinogen was found in a group of cows prior to expected date of parturition and the level of this protein in blood plasma was decreasing during lactation. The significant differences of analyzed APPs among cows before delivery, during first month after calving and in lactation (1-3 months after delivery) suggested that factors like pregnancy and stage of lactation would have an influence on their concentration.

The investigation of correlation among selected biochemical parameters and vital signs in dairy herd to design the bio-cybernetic dairy cow model.

The paper presents investigations of the relationship between the biochemical parameters and vital signs in dairy cows. We analyzed the welfare and functioning of a dairy herd using biochemical parameters and vital signs. Life and biochemical parameters were examined. In the model indicators useful for monitoring the herd are: the age of the cows, the number of cows' lactating, daily amount of received milk, length of lactation period for cows in the herd, the length of inter-calving period for cows, the number of days to effective insemination, the amount of protein in the feed, the level of ß-oxidation in leucocytes, glucose transport through red blood cells and plasma insulin. Based on the results the mathematical model was designed allowing the presentation of a cybernetic model of cow's organism. There was constructed a multi-equation model which determined the relationships between the selected variables describing the state of dairy cows in the herd and variables that characterize their welfare with its statistical verification.

Supplementation of pigs diet with zinc and copper as alternative to conventional antimicrobials.

Modern commercial pig farming systems inflict increased stress in animals, which often leads to various negative changes in the gastro-intestinal tract, especially in the case of piglets. Ban of antibiotics, used as growth promoters, has caused a need for alternatives to conventional antimicrobials in swine diets. Use of pre-/or probiotics, organic acids and plant extracts is often recommended, but it seems that zinc oxide and cooper salts, which were traditionally included in high doses to piglets diet, possess the highest efficacy. In commercial conditions feeding piglets with high doses of Zn and/or Cu stimulates piglets daily gain and decreases feed conversion factor. However, as heavy metals Zn and Cu tend to accumulate in soil and cause serious environmental pollution of soil and tap-water. Furthermore, high zinc concentrations (2500÷3000 mg/kg feed) in feed may have an impact on development of antimicrobial resistance, and may regulate the expression of genes that modify piglets' immune response. Therefore, the use of high doses of ZnO and/or Cu salts, as growth promoter, has always been a subject of discussion, and caused different legal status of such treatment in various EU countries. This short review describes current European Food Safety Authority (EFSA) point of view on the use of ZnO in medicated feed. The higher bioavailability of recently introduced new sources or forms of these metals allows for substantial reduction of dietary inclusion rate, which should have a positive outcome for pigs health and the environment.

Verapamil - L type voltage gated calcium channel inhibitor diminishes aggressive behavior in male Siamese fighting fish.

Verapamil is a L-type voltage gated calcium channels inhibitor (VGCCI), which is a highly prescribed drug used in the treatment of hypertension, angina pectoris, cardiac arrhythmia and cluster headaches. Its common use caused its appearance in water environment. VGCC inhibit epinephrine release and cause many neuro-hormonal changes influencing also fish behavior. Siamese fighting fish was chosen to study the influence of verapamil given to the water on the beginning of experiment in 3 different concentrations of 0 (control), 8 and 160 µg · L-1, on aggressive behavior in these fish. The experimental fish were placed in individual glass containers for 3 weeks and the mirror test was used. The highest concentration led to a significant modulation of fish behavior after 1 week and the lower dose caused statistically significant behavioral changes after 2 weeks of verapamil treatment. Siamese fighting fish males exposed to verapamil had longer latencies to the first chase - 12.6 s (8 µg · L-1 of verapamil) and 18.8 s (160 µg · L-1 of verapamil) compared to 5.6 s in the control group, decreased attack frequency and shorter duration of these attacks. The number of attacks within 10 min was decreased from 38.3 in the control group to 27.1 and 16.1, respectively. Also the total duration of these attacks decreased from 354.8 (control) to 326.4 (decrease statistically insignificant) and to 194.8 s in verapamil treated groups. It was shown, that even relatively low concentrations of verapamil in water may have adverse effects on fish and probably other living organisms.

IGF-I retards proper development of acinar structures formed by bovine mammary epithelial cells via sustained activation of Akt kinase.

Insulin-like growth factor-I is involved in mammary gland development, promoting proliferation and inhibiting apoptosis of mammary epithelial cells (MECs). Mitogenic actions of IGF-I are mainly mediated by the phosphatidylinositol-3 kinase (PI3K)/Akt signaling pathway. We have found that in the presence of IGF-I bovine BME-UV1 MECs cultured on reconstituted basement membrane form large spheroids with disrupted polarity and no cavity in the center. These cells showed enhanced phosphorylation of Akt, decreased level of cleaved caspase-3, and sustained proliferative activity throughout the 16-d period of 3-dimensional culture. Inhibition of the PI3K/Akt pathway by a specific inhibitor of PI3K, LY294002, resulted in the restoration of the normal acinar phenotype. However, this effect was noted only when LY294002 was added in the second week of 3-dimensional culture, which corresponded with the time of cell cycle arrest and polarity formation under control conditions. Normal development of acini was also obtained when BME-UV1 cells were treated simultaneously with IGF-I and 17ß-estradiol. The addition of 17ß-estradiol regulated Akt activation, enabling the subsequent initiation of polarization processes. 17ß-Estradiol also increased the level of IGFBP-3 protein in MECs cultured on Matrigel in the presence of IGF-I. The presented results indicate important interactions between signaling pathways activated by estrogen and IGF-I, which regulate alveologenesis in bovine mammary gland.

Effect of inulin and oligofructose enrichment of the diet on rats suffering thiamine deficiency.

Thiamine deficiency resulted in inhibition of two main pathways supplying energy to the tissues: glycolysis and ß-oxidation. Glycolysis was found to be inhibited to 40% of initial value calculated on the basis of RBC trans-membrane transport of glucose. Prolongation of experiment cause lowering of uptake of this sugar. In rats, energy production from fatty acids (FA) seems to be less sensitive to thiamine deficiency than glycolysis. After 30 days of feeding, utilization of FA in rats was depressed to the 61% of initial value. Thiamine deficiency suppressed insulin secretion, and the changes were statistically significant. Feeding of rats with thiamine restricted diet for 1 month caused the reduction of serum insulin by 14%. In the same animals, trans-membrane glucose transport was reduced over two-times, what might suggest a decreased efficiency of insulin action in such conditions. The kind and concentration of non-digestible fructo-oligosaccharides (FOS) did not affect significantly serum insulin concentration in animals fed thiamine restricted diet. Substitution of a part of wheat starch with FOS has only insignificant compensatory effect on the uptake of glucose. A partial amelioration of the ß-oxidation inhibition caused by feeding rats with thiamine deficient diet was observed in animals supplemented with FOS. However, this effect was statistically significant only in rats receiving diet containing 10% of inulin. The effect of supplemented FOS and their concentration on trans-membrane glucose transport in RBC was statistically significant, when pulled supplementation groups were used for statistical evaluation.

The influence of alpha-tocopherol supplementation on plasma concentration of this vitamin and insulin in sedentary or physically trained rats.

Male young growing rats of Wistar strain (n = 54) were fed a vitamin E deficient diet for 3 days of adaptation period and then 10 days of experimental period. After adaptation half of the rats ran, once per a day, on a treadmill with a rate of 2.0 m/s for 15 minutes for the following 10 days. Animals were given orally 0, 0.5, 1.0 or 4.0 mg/d/rat of alpha-tocopherol. The aim of the study was to evaluate the influence of physical training on tocopherol and insulin concentrations in the blood plasma of rats treated with different doses of vitamin E. The concentrations of alpha-tocopherol (HPLC-UV method used) and insulin (rat insulin specific RIA method used) were estimated in the blood plasma. Lower concentration of vitamin E in physically trained animals was observed, which may suggest increased needs of organisms for antioxidants, as a result of increased free radical production. In trained rats a lower concentration of plasma insulin was also observed. This effect is probably related to improvement in insulin action observed in organisms under physical exercise. The most significant impact on plasma alpha-tocopherol was due to its supplementation level. The treatment of rats with different doses of alpha-tocopherol was found to be not related to insulin concentration in plasma. A significant increase in feed conversion factor was noted in vitamin E treated animals. The obtained results suggest that the increased requirement of trained rats for antioxidants might be covered by alpha-tocopherol supplementation.

The influence of dietary vitamin A supplementation on vitamin A and insulin levels in sedentary or physically trained rats.

Male rats of Wistar strain (n = 48) were fed a vitamin A deficient diet for 3 days of adaptation period and then a 10 day experimental period to reduce slightly the body stores of this vitamin. Half of the animals were subjected to physical training and/or oral vitamin A supplementation. Four different doses of supplementation were used--0, 7.5, 15 and 60 microg/d/rat, which is equivalent to 0, 25, 50 and 200 IU of vitamin A, respectively. Animals from the defined groups ran on a treadmill with a rate of 2.0 m/s for 15 minutes per day for 10 days. After overnight fasting, the rats were sacrificed, and insulin in blood serum and hepatic retinol concentrations were estimated. Daily feed intake and daily body gains were similar in groups of sedentary and physically trained rats. A moderate level of oral vitamin A supplementation (the highest supplemented dose was about 6 x the above recommended NRC level) did not cause any changes in these zootechnical parameters. Oral Vitamin A supplementation resulted in an increase in retinol concentration in the liver (F = 15.2, p < 0.001), but without significant difference between trained and untrained animals. Physical training of rats caused a statistically significant decrease of insulin concentration in blood serum (1.53 +/- 0.18 vs. 1.73 +/- 0.20). This difference was highly significant (F = 11.1, p < 0.001). Vitamin A supplementation was found not to influence the concentration of this hormone, which is responsible for energy metabolism regulation in the body. Based on estimated parameters, the necessity of vitamin A excessive use in physically trained subjects was not proven.

The influence of folic acid, vitamins B(2) and B(6) supplementation on feed intake, body and organs weight, and liver fatty acids composition of rats subjected to 3 months moderate protein deprivation.

This study was conducted to determine the effect of a 3-month dietary protein restriction - protein provided 9% of energy (20% in control group). In this dietary restriction folic acid, vitamins B(2) and B(6) were delivered in amount three times above the standard level. It was observed that animals fed a protein restricted (PR) diet weighed about 5% less than animals consuming adequate diet, but the difference was not statistically significant. Enrichment of PR diet with vitamin B or folic acid caused tendency to further suppression of weight gain, and in case of vitamin B(6) these differences were statistically significant. However, such body weight (BW) suppression was not observed when all studied vitamins were used together. Significant reductions in relative liver weight (vitamin B(2) addition), the heart (folic acid) and the lungs (vitamin B(6)) were observed. The PR diet, when all vitamins were added together, caused a decrease in weights of the lungs, heart and liver scaled to BW of rats, simultaneously with a significant increase in testis weight. Feed intake and feed conversion ratio were higher in animals given PR diet without a significant influence of vitamin supplementation (except vitamin B(6) causing further increase in feed conversion ratio). Hepatic fatty acids composition of rats was not affected by protein restriction, as well as by single vitamin supplementation. However, dietary supplementation of all examined vitamins together caused a decrease in monounsaturated fatty acids followed by an increase in polyunsaturated fatty acids participation in total fatty acids pool. It seems that enrichment of PR diet with a mixture of folic acid, vitamins B(2) and B(6) resulted in a partial reverse of growth suppression and reduction in testis size in rats.

Influence of folic acid, vitamin B2 and B6 supplementation on feed intake, body and organs weight, and liver fatty acids composition in rats subjected to severe protein deprivation.

Growing rats fed for 3 months a low-protein (LP) diet (4.5% of energy from protein), possessed about 29% lower body weight than animals consuming adequate-protein diet (20% energy from protein). The LP diet feeding caused an increase in daily feed intake followed by a decrease in feed conversion efficiency. The enrichment of LP diet with folic acid, vitamin B2 and B6 (3 times above the level applied in the control diet) did not have any impact on rats BW and supplementation with these vitamins minimize the effect of LP diet on feed intake. The use of examined vitamins had a tendency to diminish an increase in feed conversion ratio caused by the LP nutrition. This effect was significant when all vitamins were added together. Rats fed the LP diet had higher relative weights of lungs, heart, liver and testis. Vitamins enriching the LP diet were observed to decrease a relative weight of lungs (folic acid, vitamin B6 and vitamin mixture), and liver (vitamin B6 and vitamin mixture). A tendency of increasing relative testis weight was also revealed in rats given the LP diet enriched with vitamins. The lower content of hepatic polyunsaturated fatty acids (FA) and a tendency for monounsaturated FA content to be higher were found in rats fed the LP diet. The LP diet enrichment with folic acid caused that these changes were more pronounced and statistically significant. Enrichment of LP diet with vitamins tested may cause a partial reverse of changes observed in the hepatic FA composition.

The background of beta-oxidation disorders in humans. I. Theory.

Disorders in energetical processes are described from the point of view of energy production and the possibility of substitution of substrates for ATP synthesis. The effects of other diet components such as microelements (Cr3+ ions) and non-digestible oligofructans (inulin and oligofructose) as stimulatory factors on the activity of beta-oxidation and cholesterol and triacylglyceroles levels lowering are reported.

The background of beta-oxidation disorders in humans. II. Laboratoy studies and clinical data.

Laboratory method for studies of clinical disorders of beta-oxidation is described. The effects of other diet components such as microelements (Cr3+ ions) and non-digestible oligofructans (inulin and oligofructose) as stimulatory factors on the activity of beta-oxidation and cholesterol and triacylglyceroles levels lowering are reported.

Diets containing leguminous seeds influence chromium content in the rat femur bone.

Leguminous seeds contain a lot of antinutritional factors (ANFs) such as protease inhibitors, lectins and condensed tannins which can affect bioavailability of nutrients. Detrimental effect of protease inhibitors can be decreased by sulphur amino acids addition. Moreover, most of the leguminous ANFs are thermolabile. Hence, legumes tested in our study were extruded and/or diets were supplemented with methionine and cystine (0.15% + 0.15%). The present experiment was performed for 28 days on 90 Wistar rats divided into nine feeding groups. Semipurified diets (10% casein) were supplemented (10%), except the control one, with soybean (S) or faba bean (F), raw (R) or extruded (E), with addition of sulphur amino acids (SAA) or without them. The Cr concentration was determined in the femur bone. Total feed intake during whole trial widely varied among the groups and was the lowest in SR (289 g) and FR (294 g) groups. There was no correlation between feed intake and Cr content in the femur bone. Generally, 10% soybean in the diet decreased Cr concentration in the bones, and faba bean increased it. The SAA addition to diets generally increased femur Cr content. However, there was a significant interaction between SAA supplementation and extrusion of faba bean, hence, the effect of their combination was unclear.

Blood plasma pseudouridine in patients with malignant proliferative diseases.

The blood plasma concentration of pseudouridine was estimated in 104 healthy adult subjects, and 108 patients suffering from malignant proliferative diseases. The HPLC method for simultaneous determination of pseudouridine and creatinine was applied. The average physiological concentration of pseudouridine in blood plasma was 2.43 +/- 0.97 mumol.l-1 or 29.15 +/- 7.40 mmol.mol-1 creatinine. The physiological urinary excretion of pseudouridine was 14.32 +/- 5.20 mumol.24 h-1.kg-0.75 or 19.60 +/- 5.22 mmol.mol-1 creatinine. Renal clearance of pseudouridine and endogenous creatinine were 4.04 +/- 0.99 and 5.50 +/- 1.46 ml.kg-0.75, respectively. A positive correlation (r = 0.55, P < 0.01) was found between age (in the range 20-92 years) and blood plasma pseudouridine concentration (mumol.l-1). By expressing plasma pseudouridine in relation to plasma creatinine, the apparent influence of non-metabolic factors (age, renal insufficiency, blood dilution) on the plasma pseudouridine concentration were largely excluded. Among haematological proliferative diseases the highest values of plasma pseudouridine concentrations were observed in chronic lymphocytic leukaemia (8.19 mumol.l-1; 54.9 mmol.mol-1 creatinine) and multiple myeloma (7.02 mumol.l-1; 52.5 mmol.mol-1 creatinine). In multiple myeloma, but not in chronic lymphocytic leukaemia, the plasma pseudouridine concentration depended on the clinical stage. A lower, but still significant response in non-Hodgkin's lymphoma was noted (4.03 mumol.l-1; 40.88 mmol.mol-1 creatinine). A significant increase of the plasma pseudouridine concentration was characteristic of adenocarcinomas of the large intestine, and it occurred in the early stages of malignant growth. In patients with lung cancer the plasma pseudouridine concentration was elevated only in advanced cases with metastases.(ABSTRACT TRUNCATED AT 250 WORDS)

The influence of age and sex on selenium distribution and glutathione peroxidase activity in plasma and erythrocytes of selenium-adequate and supplemented rats.

The experiment was performed on Sprague-Dawley male rats weighting 203, 103 and 53 g, and female 99 g. Animals were fed for 2 weeks a diet containing 0.1 and 2.0 ppm of Se (Na2SeO3 added). It was observed that the daily Se intake per kg of BW is lowered with an increase in animals body weight. Se-supplementation caused a significant increase of Se content in plasma and red blood cells. The highest concentration of Se in plasma and in RBC was found in females. GSH-Px activity was higher in RBC of all male rats receiving a Se-supplemented diet, but not in females. In plasma these differences between Se-adequate and supplemented rats were significant in youngest male rats and in females. These results suggest that age and sex of rats affect the concentration of Se and GSH-Px activity in plasma and RBC of rats.

Selenium content and distribution in bovine and human milk from different regions of Poland.

Human milk and bulk cow's milk samples from 3 different areas of central Poland were analyzed for their Se content. Cow's milk specimens were also analyzed for Se distribution and GSH-Px activity. Mature human milk contained 34-68% more Se than bulk cow's milk obtained from analogous areas of central Poland. Comparison of literature values suggest, that human and cow's milk from vicinity of Warsaw and Piotrków Trubunalski were Se-adequate but milk from Siedlce area was Se-deficient. Glutathione peroxidase activity accounted for 25-31% of the total peroxidase activity of cow's milk. The majority of the Se was found in the whey fraction. It was suggested that the participation of the casein fraction in total milk selenium increased faster than milk Se content. Cow's milk obtained near Warsaw had a 2-times higher concentration of Se than in milk from Siedlce but the content of this microelement in the casein fraction (pellet) was 3-times higher.

Selenium content and glutathione peroxidase activity of milk from vegetarian and nonvegetarian women.

The concentration of selenium (Se) in milk samples obtained from 26 lacto-ovo-vegetarian (vegetarian) women was significantly greater (22.2 +/- 0.8 ng/ml) than from 12 nonvegetarian women (16.8 +/- 1.3). Mean GSH-Px activity (EC 1.11.1.9) in milk from vegetarians was 146% of that in milk from nonvegetarians. A significant correlation between GSH-Px activity and Se concentration was observed (r = 0.76). Likewise a significant correlation was observed between milk linoleic acid content and GSH-Px activity (r = 0.68). In undialyzed samples from vegetarian women, most of the increased content of Se was found in fractions containing proteins of 100 kdaltons (kD) or more. The high glutathione peroxidase activity in milk from vegetarians was associated with selenoproteins in the 90 to 100 kD range. A distinct 40 kD Se peak was also detected in milk from vegetarians. There was no difference in Se intake between the two groups of women. Therefore, although data from the present study show that both the quantity and distribution of Se in human milk can be modified by the maternal diet, the increased Se content and GSH-Px activity in milk from vegetarians cannot be explained by an increased Se intake.

Selenium content and distribution of human, cow and goat milk.

Studies were undertaken to compare the content and distribution of selenium in human, cow and goat milk. Selenium content of cow milk was found to be lower than that of either human or goat milk. Regardless of source, less than 3% of total milk selenium was associated with the lipid fraction. Selenium within the 120,000 X g supernatant accounted for 72, 62 and 30% of the total in cow, human and goat milk, respectively. Glutathione peroxidase occurred in all milk samples with goat greater than human greater than cow. Percent of total peroxidase activity associated with glutathione peroxidase was 29, 27 and 65 for human, cow and goat milk, respectively. Approximately 20-28% of the selenium in milk was removed by dialysis (molecular exclusion of 6-8 kDa). After gel chromatography, 8-12 selenoprotein fractions were detected in undialyzed skim milk from each species. Most of the glutathione peroxidase activity was found in the fractions corresponding to 170 and 96 kDa in milk from all species examined. The diameric form of glutathione peroxidase also appeared in dialyzed and undialyzed milk. Distinct differences in the content and distribution of selenoproteins among these species in fresh and dialyzed milk are discussed.