Ewa Tomaszewska, Piotr Dobrowolski, Małgorzata Kwiecień, Agata Wawrzyniak and Natalia Burmańczuk
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The aim of the study was to investigate gender-specific effects of a maternal treatment with dexamethasone (DEX) at multiple maximum therapeutic doses during the last 25 d of pregnancy on the bone tissue metabolism in neonatal piglets. BMD and BMC were measured. The geometry, histomorphometry, and mechanical properties of bones were determined. The assessment of selected hormones, cytokines, and amino acids was performed in 35-day-old piglets. DEX administered prenatally inhibited neonatal bone development, as well as bone mineral density, and geometrical and mechanical properties of bones. Neonatal GC-induced bone loss was linked with disturbed somatotropic axis function and decreased serum concentration of GH in male piglets and decreased concentration of IGF-1 in females. Prenatal DEX treatment led to an increase in the concentration of insulin and enhanced activity of bone specific alkaline phosphatase in piglets of both genders.
Ewa Tomaszewska, Piotr Dobrowolski, Małgorzata Kwiecień, Natalia Burmańczuk, Barbara Badzian, Sylwia Szymańczyk and Paulina Kurlak
The aim of this study was to define the effects of diet containing the same mineral content of mineral salt or amino acid chelate, and diet containing various levels of Cu amino acid chelate on liver histomorphometry in growing rats. Male Wistar rats were used in the 12th week experiment. The control group (n = 12) was fed standard diet, which provided Cu in an inorganic form at the level required for rats. The experimental animals were divided into four groups (each n = 12) depending on different levels (100%, 75%, 50%, 25% covered daily demand) of Cu supplementation in chelated form. Cu content was determined in the liver tissue and blood plasma. Immunohistochemical staining with caspase-3 antibody was performed. Microscopic assessment of the liver structure indicated that Cu supplementation did not change the liver architecture. However, histomorphometric analysis revealed a significant increase in the number of nuclei, total cell number, and multinucleated hepatocytes in rats supplemented with the organic form of Cu at the level of 25% compared with the control group. There was a considerable increase in the number of apoptotic cells and ballooning degeneration of hepatocytes, especially in groups supplemented with organic form of Cu covering the daily demand in 100% and 75%, in comparison to control group. Moreover, there was no Cu deposition in the liver and changes in Cu content in blood. Cu provided in the diet in organic form covering an amount of its minimum daily demand in 25% appears to be the least harmful with regard to the liver. It indicates that there is a need to establish the level of diet supplementation with Cu amino acid chelates.
Ewa Tomaszewska, Piotr Dobrowolski and Małgorzata Kwiecień
Copper (Cu) is required for all basic biochemical and physiological processes. The aim of this study was to evaluate the effects of different sources of dietary Cu on the histomorphometry of liver and jejunal epithelium in adult rats. Male 12-week-old rats were used in a 12-week experiment. The control diet provided the required Cu level from sulfate, and other two diets were supplemented with Cu as a glycine complex at 75% and 100% of daily requirement. Basal hematological and plasma biochemical analyses were also performed. There was no effect of Cu supplementation on the liver weight and the plasma and liver Cu concentration. Histomorphometric analysis of liver tissue showed an increase in the collagen amount and intracellular space in the group supplemented with Cu amino acid. Cu given in the organic form at 100% of daily requirement decreased the muscular and submucosa layer and the crypt depth. In turn, organic copper given at 75% of daily requirement did not influence the intestinal morphology. Dietary Cu given to adult rats as copper sulfate or a glycine complex meeting 100% of the daily requirement appears to be less harmful with regard to intestinal epithelium than when given as a glycine complex at 100% of daily requirement.
Ewa Tomaszewska, Siemowit Muszyński, Piotr Dobrowolski, Małgorzata Kwiecień, Renata Klebaniuk, Sylwia Szymańczyk, Agnieszka Tomczyk, Sylwester Kowalik, Anna Milczarek and Izabela Świetlicka
Faba bean (FB) seeds can be a good protein-energy component in animal feed. However, the presence of anti-nutritional substances is a negative feature of FB seeds. The aim of this study was to examine the influence of different levels of unprocessed FB seeds in feed on the gut-bone axis and metabolic profile in broilers. Ninety six, 1-day-old Ross 308 broiler chickens were randomly selected to one of the 3 dietary treatments (32 chickens in each, divided into 8 pens with 4 birds per each pen): the control group fed standard diet with soybean meal and without FB seeds, group I fed 8/15% (starter/grower) of high-tannin FB seeds, and group II fed 16/22% of high-tannin FB seeds. Bone mechanical examination, hematological and serum biochemical analysis as well histomorphometry of small intestine and liver tissue were performed. The intake of high-tannin FB seeds, irrespective of their amount, did not alter the bone geometric, mechanical and densitometric parameters nor influenced basal hematological parameters, however it resulted in: decreased serum concentration of total cholesterol and calcium; a reduced longitudinal myenteron of small intestine; increased mucosa and villus epithelium thickness, villus length, thickness and absorptive surface in duodenum; increased number of active crypts in jejunum; unchanged collagen area, intercellular space, and total cell number in the liver; decreased number of multinuclear hepatocyte cells. Moreover, the livers of birds fed the higher dose of high-tannin FB seeds had lymphocytic infiltrates in portal tracts and sinusoids. Feeding of unprocessed high-tannin FB seeds exerted an influence on the gastrointestinal tract by increased absorptive surface. In conclusion, the dietary inclusion of unprocessed high-tannin FB seeds had no negative effects on broiler growth, tibial bone mechanical properties and intestinal characteristics. Unprocessed high-tannin FB seeds may be used in broiler diets, but their dietary levels should not be higher than those discussed.
Katarzyna Mehr, Paweł Piotrowski, Bogumiła Frączak and Maja Matthews-Kozanecka
The aim of the study was to compare erupted permanent bovine teeth of two types involved in the process of chewing in respect to the localisation and histomorphometric characteristics of enamel tufts. The research material comprised 240 fully erupted premolars and molars from the maxillae and mandibles of 27 heads of cattle from the Polish Black-and-White breed. Overall, 1,986 specimens of bovine teeth were analysed using a Nikon Eclipse E600 microscope with Micro Image v4.0 software. Enamel tufts were relatively numerous in the enamel of smooth surfaces - on average from 5.6 to 6.4 per cross-section of tooth crown. The average length of the enamel tufts expressed by means of a median was smaller for premolars (89.3 μm) than for molars (123.9 μm). The analysis of the value of Spearman's rank correlation coefficient revealed that no relationship existed between the length and width of tufts in molars (rs =0.26), and a low-level relationship was noted (rs =0.45) in premolars. On the basis of the Mann-Whitney U test for the metric measurements, no significant differences were discovered for the width of enamel tufts (P=0.427), but significant differences were observed for the length of enamel tufts (P=0.032). The observed differences in the histological structure of the enamel of posterior bovine teeth in comparison to human teeth indicate that a certain degree of caution ought to be considered when using bovine teeth as a substitute for human teeth in in vitro trials.
Halyna Rudyk, Ewa Tomaszewska, Ihor Kotsyumbas, Siemowit Muszyński, Agnieszka Tomczyk-Warunek, Sylwia Szymańczyk, Piotr Dobrowolski, Dariusz Wiącek, Daniel Kamiński and Oksana Brezvyn
Fumonisins are strongly toxic metabolites of Fusarium proliferatum and Fusarium verticillioides commonly present in corn-based feed. The aim of the study was to evaluate bone homeostasis in experimental fumonisins B1 and B2 intoxication of rats, a vertebrate animal model of toxicological studies, as still little is known about the possible disturbing effect of fumonisins on bone homeostasis. Adolescent (5-week-old) male Wistar rats were randomly assigned into a control group and a group FB intoxicated with fumonisins by daily intragastric administration of fumonisins at the dose of 90 mg/kg of body weight per animal in the FB group for 21 days. The fumonisin intoxication did not affect body and bone mass, although the mechanical and geometric properties were decreased in fumonisin-intoxicated rats. Bone volumetric and mineral density did not differ between groups, but bone mineral content and bone ash percentage was lower in the FB group. Detailed analysis showed that Ca, Cu, Fe, Mn, Sr, and Zn bone content significantly decreased in fumonisin intoxicated rats and the alterations in structure of bone mineral phase (reduction of the apatite-bone crystals size) were noted. While the negative structural alterations in growth plate and articular cartilages were also observed, fumonisin intoxication improved histomorphometrical parameters of trabecular bone. Concluding, the dose of fumonisins used in the present study caused hepatotoxic effect, which was sufficient to trigger the disturbance in mineral homeostasis resulting in altered bone metabolism and decreased mechanical endurance.
Florina Popovska-Perčinić, Ivana Jarić, Lazo Pendovski, Ristić Nataša, Svetlana Trifunović, Verica Milošević and Vladimir Ajdžanović
In areas with moderate continental climate, increased average ambient temperature during the summer represents a stressogenic factor that affects the hypothalamo-pituitaryadrenocortical axis in mammals. Therefore, we wanted to examine the effects of 4 days of constant exposure to moderately elevated ambient temperature (35 ± 1oC) on the histomorphometric and immunofl uorescent characteristics, as well as on the hormonal secretion of pituitary corticotropes (ACTH) cells in adult male rats. In comparison with the controls kept at 20 ± 2oC, a signifi cant increase (p<0.05) of the absolute and relative pituitary weight (23.1% and 36.1%, respectively) was registered after exposure to heat. The localization, as well as the shape of the ACTH cells in the heat exposed group was not signifi cantly altered, but their immunopositivity was weaker. After 4 days of heat exposure, a weaker signal confi rmed the relative fl uorescence intensity of the ACTH cells (15.3%, p<0.05). In heat exposed rats, an increase of the cellular and nuclear volumes of immunolabelled ACTH cells and decrease of their volume density (6.9%, 14.3% and 20.0%, respectively; p<0.05) was registered. Observed histomorphometric and immunofl uorescent features of the pituitary ACTH cells were in accordance with the increased (p<0.05) value of plasma adrenocorticotropic hormone (ACTH) by 23.7% compared to the control rats. It can be concluded that the 4-day exposure to moderately elevated ambient temperature intensifi es pituitary ACTH secretion in adult male rats.
Ewa Tomaszewska, Piotr Dobrowolski, Małgorzata Kwiecień, Anna Winiarska-Mieczan, Agnieszka Tomczyk, Siemowit Muszyński and Bożena Gładyszewska
Administration of the amino acid copper (Cu) complex ensures higher Cu bioavailability through enhanced absorption from intestine and decreases the dietary Cu level, compared to the recommended Cu dose. The objective of this study was to investigate the effect of Cu-low diet on the bone development in adolescent rats. Male rats at the age of 6 weeks were used in the 12-week experiment. The control diet provided the required Cu level from sulfate (S-Cu) and other diets were supplemented with Cu as a glycine complex (Cu-Gly) at 25%, 50%, 75%, and 100% of daily requirement. After the 12-week treatment, rats from the Cu-Gly100 group were heavier, compared to the other groups. The copper and calcium plasma and bone concentrations of the rats in the groups treated with the organic form of Cu (irrespective of its dose) was similar to the control values noted in the rats administered with S-Cu. A decrease in the femur weight and length was observed in the Cu-Gly75 and Cu-Gly50 groups. Cu-Gly increased the cross section area, mean relative wall thickness, and cortical index only in the Cu-Gly75 group. A decrease in the ultimate strength, elastic stress, and ultimate stress was noted in the Cu-Gly100 and Cu-Gly75 groups. In the Cu-Gly50 group, a decrease in the ultimate stress and an increase in the maximal elastic strength and bending moment were noted. Adolescent rats treated with Cu-Gly at a Cu-deficient level exhibited a dose-dependent strongly osteoporotic cancellous bone. Lower proteoglycan content was found in groups fed the Cu-low diet. In the control rats supplemented with S-Cu, there was no evident gradient in safranin O staining. It is difficult to indicate which dose of the Cu-Gly complex among the investigated Cu-poor diet exerted a positive effect on bone metabolism. It appears that the use of this Cu-Gly complex at a significantly reduced dose than S-Cu at the recommended dose did not inhibit the development of bone and hyaline cartilage in adolescent rats.
Barbara Mnich, Janusz Skrzat and Krzysztof Szostek
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