Innovative high fat diet establishes a novel zebrafish model for the study of visceral obesity.

Obesity is a complex chronic condition associated with multiple health risks, including visceral obesity, which is particularly detrimental. To gain insight into the mechanisms underlying obesity and its associated pathologies, a novel zebrafish model was established using an innovative high-fat diet (HFD). The primary goal was to induce visceral obesity in zebrafish and study the associated structural changes. To achieve this, a unique HFD consisting of 40% beef fat (HFD40) was developed and supplemented with magnesium aluminometasilicate to improve stability in a high humidity environment. Feeding regimens were initiated for both juvenile (starting at 2 weeks post-fertilization, lasting 18 weeks) and adult zebrafish (3 months post-fertilization, 8 weeks feeding duration). The innovative dietary approach successfully induced visceral obesity in both juvenile and adult zebrafish. This new model provides a valuable tool to study obesity-related pathologies, metabolic syndrome, and potential therapeutic interventions. Most importantly, the low-cost and easy-to-prepare composition of HFD40 was seamlessly incorporated into the water without the need for separation, was readily absorbed by the fish and induced rapid weight gain in the zebrafish population. In conclusion, this study presents a novel HFD40 composition enriched with a high beef fat concentration (40%), which represents a significant advance in the development of an experimental zebrafish model for the study of visceral obesity and associated metabolic changes.

Effect of Microbial Phytase on Ileal Digestibility of Minerals, Plasma and Urine Metabolites, and Bone Mineral Concentrations in Growing-Finishing Pigs.

The study was conducted to evaluate the effects of added phytase in the diet of pigs on utilization of minerals and to determine the essential levels of this supplement in compound feed. An additional aim of the study was a critical assessment of current recommendations regarding the level of phosphorus in the diet of pigs, taking into account the use of phytase. A total of 432 pigs were allotted to six dietary treatments, with nine replicate pens per treatment according to body weight (BW) and sex. The treatments included a negative control (NC) with reduced content of digestible phosphorus; the NC diet supplemented with 6-phytase produced by a genetically modified strain of Aspergillus oryzae in the amount of 250 (NC + 250), 500 (NC + 500), 1000 (NC + 1000), or 1500 (NC + 1500) FTU/kg of feed; and a positive control (PC) diet formulated to meet NCR nutrient requirements for pigs. The results showed that, the higher the phytase activity in the diet (NC+), the lower the concentration of P, Ca, and Mg in the chyme (p < 0.05). Pigs fed the PC and NC+ diets had higher plasma levels of P and Ca than group NC in both fattening periods (p < 0.05). The content of phosphorus in the femur of pigs increased with the level of phytase added to the diet (p < 0.05). The content of Cu and Zn in the femur of pigs in the growing period was higher in groups NC + 500 and NC + 1000 than in the remaining experimental groups (p < 0.001). The content of P and Ca in the feces was higher in pigs fed the PC diet in comparison to the remaining experimental groups in both fattening periods (p < 0.001). There was a decrease in the content of P, Ca, and Mg in the excreta of pigs fed NC+ diets in both fattening periods (p < 0.05). A linear decrease in excretion of zinc in the feces was noted in the case of high levels of phytase, i.e., 1000 and 1500 FTU (p < 0.001). Increasing the level of phytase decreased the Cu (p < 0.001) content in the urine of growing−finishing pigs. In conclusion, the analysis of the effects of the use of phytase in a range of 0−1500 FTU/kg in low-phosphorus diets for fattening pigs indicates that 1000 FTU is the most effective level of phytase for increasing utilization of minerals and reducing excretion of elements into the environment.

The Impact of Herbal Additives for Poultry Feed on the Fatty Acid Profile of Meat.

Researchers often found that herbal additives to chicken feed can favorably alter the fatty acid profile of the meat. The most desirable effects of diet modification comprise an increased content of polyunsaturated fatty acids (PUFA) and monounsaturated fatty acids (MUFA) and a reduced content of saturated fatty acids (SFA) in the breast and thigh muscles. A modified fatty acid profile contributes to improvement in the quality of poultry meat, which is reflected in its increased consumption. However, it may be problematic that PUFAs are oxidized easier than other lipids, which can have a negative impact on the sensory traits of meat. By contrast, herbs and herbal products contain antioxidants that can prevent the oxidation of unsaturated fatty acids and cholesterol present in animal-origin products and increase the antioxidant potential of the consumer's body. This paper aims to review the influence of herbal additives for broiler chicken diets on the fatty acid profile of poultry meat. Special attention was paid to changes in the content of SFAs, MUFAs, and PUFAs, but also alterations in the omega-6:omega-3 ratio. The presented reference literature supports the statement that herbs and bioactive components of herbs added to chicken diets can improve the quality of broiler chicken meat by altering the content of fatty acids.

Dietary Alpha-Ketoglutarate Partially Abolishes Adverse Changes in the Small Intestine after Gastric Bypass Surgery in a Rat Model.

Alpha-ketoglutarate (AKG) is one of the key metabolites that play a crucial role in cellular energy metabolism. Bariatric surgery is a life-saving procedure, but it carries many gastrointestinal side effects. The present study investigated the beneficial effects of dietary AKG on the structure, integrity, and absorption surface of the small intestine after bariatric surgery. Male 7-week-old Sprague Dowley rats underwent gastric bypass surgery, after which they received AKG, 0.2 g/kg body weight/day, administered in drinking water for 6 weeks. Changes in small intestinal morphology, including histomorphometric parameters of enteric plexuses, immunolocalization of claudin 3, MarvelD3, occludin and zonula ocludens 1 in the intestinal mucosa, and selected hormones, were evaluated. Proliferation, mucosal and submucosal thickness, number of intestinal villi and Paneth cells, and depth of crypts were increased; however, crypt activity, the absorption surface, the expression of claudin 3, MarvelD3, occludin and zonula ocludens 1 in the intestinal epithelium were decreased after gastric bypass surgery. Alpha-ketoglutarate supplementation partially improved intestinal structural parameters and epithelial integrity in rats undergoing this surgical procedure. Dietary AKG can abolish adverse functional changes in the intestinal mucosa, enteric nervous system, hormonal response, and maintenance of the intestinal barrier that occurred after gastric bypass surgery.

The Effect of L-Glutamine on Basal Albumen and Yolk Indices, and Albumen Amino Acids Composition.

The current study tested the hypothesis that 1.0% dietary inclusion of L-glutamine (Gln), an non-essential amino acid that influences protein synthesis, can improve internal egg quality, including amino acids profile. Thirty-week-old Bovans Brown laying hens in their middle laying period were assigned to one of the two experimental groups (12 replicate cages, 2 hens/cage) with Gln in the form of alpha-ketoglutarate (10 g/kg) or without Gln inclusion. The experimental period lasted for 30 wks, from the 31st to the 60th week of age of hens, when eggs were collected and selected egg quality indices were determined. Gln supplementation had no effect on albumen and egg yolk share, albumen and yolk basal indices and composition, including yolk cholesterol content. However, Gln decreased the lipid content of the egg albumen (p < 0.001), and influenced albumen amino acid profile, increasing content of asparagine (p < 0.05), phenylalanine (p < 0.05), proline (p < 0.001), tryptophan (p < 0.01), and tyrosine (p < 0.05). In conclusion, the study shows a potential role of Gln supplementation for enhancing nutritional values of eggs by lower lipid content and higher amino acid profile.

Antioxidant, Anti-Inflammatory, and Immunomodulatory Properties of Tea-The Positive Impact of Tea Consumption on Patients with Autoimmune Diabetes.

The physiological markers of autoimmune diabetes include functional disorders of the antioxidative system as well as progressing inflammation and the presence of autoantibodies. Even though people with type 1 diabetes show genetic predispositions facilitating the onset of the disease, it is believed that dietary factors can stimulate the initiation and progression of the disease. This paper analyses the possibility of using tea as an element of diet therapy in the treatment of type 1 diabetes. Based on information available in literature covering the last 10 years, the impact of regular tea consumption or diet supplements containing tea polyphenols on the oxidative status as well as inflammatory and autoimmune response of the organism was analyzed. Studies conducted on laboratory animals, human patients, and in vitro revealed positive effects of the consumption of tea or polyphenols isolated therefrom on the diabetic body. Few reports available in the literature pertain to the impact of tea on organisms affected by type 1 diabetes as most (over 85%) have focused on cases of type 2 diabetes. It has been concluded that by introducing tea into the diet, it is possible to alleviate some of the consequences of oxidative stress and inflammation, thus limiting their destructive impact on the patients' organisms, consequently improving their quality of life, regardless of the type of diabetes. Furthermore, elimination of inflammation should reduce the incidence of immune response. One should consider more widespread promotion of tea consumption by individuals genetically predisposed to diabetes, especially considering the drink's low price, easy availability, overall benefits to human health, and above all, the fact that it can be safely used over extended periods of time, regardless of the patient's age.

Fourier Transform Infrared Microspectroscopy Combined with Principal Component Analysis and Artificial Neural Networks for the Study of the Effect of ß-Hydroxy-ß-Methylbutyrate (HMB) Supplementation on Articular Cartilage.

The potential of Fourier Transform infrared microspectroscopy (FTIR microspectroscopy) and multivariate analyses were applied for the classification of the frequency ranges responsible for the distribution changes of the main components of articular cartilage (AC) that occur during dietary ß-hydroxy-ß-methyl butyrate (HMB) supplementation. The FTIR imaging analysis of histological AC sections originating from 35-day old male piglets showed the change in the collagen and proteoglycan contents of the HMB-supplemented group compared to the control. The relative amount of collagen content in the superficial zone increased by more than 23% and in the middle zone by about 17%, while no changes in the deep zone were observed compared to the control group. Considering proteoglycans content, a significant increase was registered in the middle and deep zones, respectively; 62% and 52% compared to the control. AFM nanoindentation measurements collected from animals administered with HMB displayed an increase in AC tissue stiffness by detecting a higher value of Young's modulus in all investigated AC zones. We demonstrated that principal component analysis and artificial neural networks could be trained with spectral information to distinguish AC histological sections and the group under study accurately. This work may support the use and effectiveness of FTIR imaging combined with multivariate analyses as a quantitative alternative to traditional collagenous tissue-related histology.

ß-Hydroxy-ß-Methylbutyrate (HMB) Supplementation Prevents Bone Loss during Pregnancy-Novel Evidence from a Spiny Mouse (Acomys cahirinus) Model.

The aim of this study was to determine the effects of ß-hydroxy-ß-methylbutyrate (HMB) supplementation during pregnancy on postpartum bone tissue quality by assessing changes in trabecular and compact bone as well as in hyaline and epiphyseal cartilage. The experiment was carried out on adult 6-month-old female spiny mice (Acomys cahirinus) divided into three groups: pregnant control (PregCont), pregnant HMB-treated (supplemented with 0.02 g/kg b.w of HMB during the second trimester of pregnancy, PregHMB), and non-pregnant females (NonPreg). Cross-sectional area and cortical index of the femoral mid-shaft, stiffness, and Young modulus were significantly greater in the PregHMB group. Whole-bone mineral density was similar in all groups, and HMB supplementation increased trabecular number. Growth plate cartilage was the thinnest, while the articular cartilage was the thickest in the PregHMB group. HMB supplementation increased the content of proteoglycans in the articular cartilage and the percentage of immature collagen content in metaphyseal trabeculae and compact bone. In summary, dietary HMB supplementation during the second trimester of pregnancy intensifies bone metabolic processes and prevents bone loss during pregnancy.

The effect of maternal HMB supplementation on bone mechanical and geometrical properties, as well as histomorphometry and immunolocalization of VEGF, TIMP2, MMP13, BMP2 in the bone and cartilage tissue of the humerus of their newborn piglets.

The presented experiment focuses on assessing the impact of HMB (hydroxy-ß-methobutyrate) supplementation of mothers during pregnancy on the development of the skeletal system of their offspring. For this purpose, an experiment was carried out on 12 clinically healthy sows of the Great White Poland breed, which were divided randomly into two groups the control and the HMB group. All animals were kept under standard conditions and received the same feed for pregnant females. In contrast, females from the HMB group between 70 and 90 days were supplemented with 3-hydroxy-3-methylbutyle in the amount of 0.2g/kg b.w/day. Immediately after birth, the piglets were also divided into groups based on: sex, and presence or lack HMB supplementation, and subsequently were euthanized and humerus bones from all piglets were collected. Mother's HMB supplementation during pregnancy affected the multiple index of their offspring. The higher humerus mass and length was observed with the greater effect in males. Maternal supplementation also influenced on the geometrical and mechanical properties of the humerus as in the case of mass, this effect was higher in males. Also, the collagen structure of the compacted and trabecular bone changed under the HMB addition. Maternal supplementation also affected the expression of selected proteins in growth cartilage and trabecular bone. The obtained results show that the administration to the mother during pregnancy by the HMB significantly affects the development of the humerus in many ways. The obtained results also confirm the utility of such experiments in understanding of the importance of the pregnancy diet as an develop and adaptable factor of offspring organisms and are the base for further research in that area as well as in the protein markers expression area.

The Effect of Dietary Rye Inclusion and Xylanase Supplementation on Structural Organization of Bone Constitutive Phases in Laying Hens Fed a Wheat-Corn Diet.

This study was conducted to examine the effect of dietary rye inclusion and xylanase supplementation on the bone quality of ISA Brown laying hens. Ninety-six laying hens were assigned to four groups: fed with wheat-corn diet or rye-wheat-corn diet (25% of hybrid rye inclusion) or nonsupplemented or supplemented with xylanase (200 mg/kg of feed) for a period of 25 weeks, from the 26th to the 50th week of age. X-ray absorptiometry, X-ray diffraction, and Fourier-transform infrared spectroscopy were used to provide comprehensive information about the structural organization of bone constitutive phases of the tibia mid-diaphysis in hens from all treatment groups. Bone hydroxyapatite size was not affected by diet. Xylanase supplementation influenced the carbonate-to-phosphate ratio and crystallinity index in hens fed with both diets. Xylanase had more pronounced effects on bone mineral density and collagen maturity in hens fed with the rye-wheat-corn diet versus those fed with the wheat-corn diet. The results of this study showed that modern rye varieties, when supplemented with exogenous xylanase, can be introduced to the diet of laying hens without any adverse effects on bone structure.

Influence of Phytase Supplementation at Increasing Doses from 0 to 1500 FTU/kg on Growth Performance, Nutrient Digestibility, and Bone Status in Grower-Finisher Pigs Fed Phosphorus-Deficient Diets.

The objective of the current study is to assess the effects of the inclusion of 6-n phytase to a phosphorous-deficient diet on the growth performance (feed intake, average daily gain, and feed conversion ratio), apparent digestibility of calcium and phosphorus, and bone characteristics of grower-finisher pigs. The experimental diets included a phosphorus-deficient diet containing 0 (negative control), 250, 500, 1000, or 1500 FTU/kg of 6-phytase, and a diet formulated to meet the phosphorus nutrient requirements of pigs (positive control). Pigs were fed the experimental diets from the time they were ~35 kg body weight until they reached slaughter weight of ~110 kg. Bone status of the metacarpal (ash, mineral content) and femur (mineralization, geometry, and mechanical strength) bones were assessed. There was no effect of dietary treatment on feed intake. Feed conversion ratio was improved following inclusion of phytase at a dose of 500 FTU/kg or higher. Phytase inclusion at a dose of 1000 FTU/kg increased the average daily weight gain of grower-finisher pigs. Phytase inclusion at a dose of 500 FTU/kg was sufficient to increase metacarpal phosphorus content. Femur mid-diaphysis ash percentage was significantly increased even after the inclusion of the lowest dose of phytase. Analysis of structural parameters of femur mechanical strength (Young's modulus, yield stress, yield strain, ultimate stress, ultimate strain) showed that the inclusion of a phytase dose of 500 FTU/kg in growing/finishing diets was sufficient to significantly improve bone status of grower-finisher pigs at slaughter.

Is Dietary 2-Oxoglutaric Acid Effective in Accelerating Bone Growth and Development in Experimentally-Induced Intrauterine Growth Retarded Gilts?

In this study, the effect of long-term 2-oxoglutaric acid (2-Ox) supplementation to experimentally-induced intrauterine growth retarded gilts was examined. Sows were treated with synthetic glucocorticoid (dexamethasone) every second day, during the last 45 days of pregnancy, at a dose of 0.03 mg/kg b.w. At birth, the gilts were randomly divided into two groups: unsupplemented and supplemented with 2-Ox for nine months (0.4 g/kg body weight/day). Oral supplementation of 2-Ox to experimentally-induced intrauterine growth retarded gilts increased body weight at weaning as well as final body weight at the age of nine months, and showed a regenerative effect on bone mineralization and morphology of trabeculae and articular cartilage. The positive effects on bone structure were attributed to the 2-Ox-induced alterations in bone metabolism, as evidenced by the changes in the expression of proteins involved in bone formation and remodeling: osteocalcin (OC), osteoprotegerin (OPG), receptor activator of nuclear factor kappa-Β ligand (RANKL), tissue inhibitor of metalloproteinases 2 (TIMP-2), bone morphogenetic protein 2 (BMP-2), cartilage oligomeric matrix protein (COMP), and vascular endothelial growth factor (VEGF).

The effect of bee pollen on bone biomechanical strength and trabecular bone histomorphometry in tibia of young Japanese quail (Coturnix japonica).

It has been demonstrated in numerous studies that bee pollen supplementation shows numerous positive effects on health. However, its impact on bones is largely unknown. The purpose of this study was to investigate the effect of bee pollen supplementation on the tibia biomechanical properties and bone morphometric measures using Japanese quail as an animal model. The experiment was arranged in a 2x2x2 factorial design, with sex, quail line (meat-type or egg-lying type), and bee pollen inclusion (0 or 10 g/kg of feed) as factors. The quails were one-day-old at the beginning of the experiment, they were euthanized after 42 days. Our study showed for the first time unfavorable effects of bee pollen on bones properties. Bee pollen supplementation negatively affected bone structure, irrespective of quails' sex or line type. Bone length (P < 0.001), weight (P < 0.01), and mean relative wall thickness (P < 0.01) and mineralization (P < 0.05) were reduced by bee pollen treatment. For female quails, irrespective of line type, the decrease of yield load (P < 0.001), ultimate load (P < 0.01), yield stress (P < 0.001) and ultimate stress (P < 0.05) was noted. Analysis of growth plate in bone metaphysis showed that bee pollen supplementation slowed the process of bone maturation irrespective of sex (P < 0.05). On contrary, dietary bee pollen positively affected bone homeostasis of trabecular bone in bone metaphysis as bone mineral density increased in experimental groups (P < 0.05). In males, this was the result of the increase of trabecular thickness (P < 0.01), in females due to the reduction of trabecular space (P < 0.001). In conclusion, our results demonstrate that bee pollen (1.0%, 10 g/kg of feed) supplementation caused significant negative effects on the mechanical endurance of the tibia of quails, while showed beneficial effects on trabecular bone histomorphometry.

Chronic dietary supplementation with kynurenic acid, a neuroactive metabolite of tryptophan, decreased body weight without negative influence on densitometry and mandibular bone biomechanical endurance in young rats.

Kynurenic acid (KYNA) is a neuroactive metabolite of tryptophan. KYNA naturally occurs in breast milk and its content increases with lactation, indicating the role of neonatal nutrition in general growth with long-term health effects. KYNA is also an antagonist of ionotropic glutamate receptors expressed in bone cells. The aim of this study was to establish the effects of chronic KYNA supplementation on bone homeostasis in young rats, using mandible as a model bone. Female and male newborn Wistar rats were divided into control and KYNA-administered groups until 60 days of age (25x101 mg/L or 25x102 mg/L in drinking water). Hemimandibles were subjected to densitometry, computed tomography analysis and mechanical testing. Rats supplemented with KYNA at both doses showed a decrease in body weight. There were no effects of KYNA administration and mandible histomorphometry. In males, a significant quadratic effect (P < 0.001) was observed in the densitometry of the hemimandible, where BMD increased in the group supplemented with 2.5x101 mg/L of KYNA. Analysis of mechanical tests data showed that when fracture forces were corrected for bone geometry and rats body weight the improvement of bone material properties was observed in male and female rats supplemented with lower dose of KYNA. This study showed that chronic supplementation with KYNA may limit weight gain in the young, without adversely affecting the development of the skeleton.

Surface analysis of etched enamel modified during the prenatal period.

Structural changes in the enamel surface subjected to induced demineralization and assessment of the influence of prenatal administration of ß-hydroxy ß-methylbutyrate (HMB) on enamel resistance were investigated. The examination was conducted on five sets of teeth from one-day-old spiny mice (Acomys cahirinus), one from the control and four from the experimental groups. Surface structure, molecular arrangement and crystalline organization of offspring's enamel both before and after etching were studied. Obtained results revealed that the physical and molecular arrangements of enamel were altered after the prenatal supplementation, and significantly affected its final structure and resistance against acid action. The enamel of incisors from the offspring which mothers were supplemented with HMB in a high dose (0.2 g/kgbw) and in the late period of gestation (26th-39th day) showed the highest endurance against acid treatment demonstrating only vestigial changes in their surface structure after acid action. Comparing to the remaining experimental groups, it was characterized by a reduced roughness and fractal dimension, significantly lower degree of demineralization and simultaneous lack of notable differences in the Raman spectra before and after acid etching. The results suggest that an increased enamel resiliency was the effect of a relatively high degree of mineralization and higher organization of the surface.

The Influence of the Partial Replacing of Inorganic Salts of Calcium, Zinc, Iron, and Copper with Amino Acid Complexes on Bone Development in Male Pheasants from Aviary Breeding.

This study analyzed the effects of partial replacing of Ca, Fe, Zn, and Cu salts with glycine chelates on the measures of bones health in 16-week-old captive-reared male pheasants, allocated to one of the three experimental groups supplemented with Ca, Fe, Zn, and Cu in forms of inorganic salts (the control group) or groups receiving from the ninth week 25% and 50% of supplemented elements as glycine chelates. At the end of rearing birds receiving chelates were heavier (p < 0.001) and their tibia showed an increase of numerous mechanical parameters: yield and ultimate force (p = 0.028, p < 0.001, respectively), stiffness (p = 0.007), Young modulus (p < 0.001), compared to the control animals. The bones of birds receiving chelates in 50% were also heavier (p < 0.001) and longer (p = 0.014), with thinner cortical bone in midshaft (p = 0.027) and thicker proximal trabeculae (p < 0.001) compared to the control. While both doses of chelates increased mineral density in midshaft (p = 0.040), bone content of Cu and Zn decreased (p = 0.025, p < 0.001, respectively). The content of immature collagen in cancellous bone and articular cartilage increased in groups receiving chelates (p < 0.001, p = 0.001, respectively). In conclusion, glycine chelates probably enhanced development of the skeletal system in male pheasants as bones were denser and more resistant to mechanical damage.

Kynurenic acid as the neglected ingredient of commercial baby formulas.

The global increase in resorting to artificial nutritional formulas replacing breastfeeding has been identified among the complex causes of the obesity epidemic in infants and children. One of the factors recently recognized to influence metabolism and weight gain is kynurenic acid (KYNA), an agonist of G protein-coupled receptor (GPR35). Therefore the aim of the study was to determine the concentration of KYNA in artificial nutritional formulas in comparison with its level in human breast milk and to evaluate developmental changes in rats exposed to KYNA enriched diet during the time of breastfeeding. KYNA levels were measured in milk samples from 25 heathy breast-feeding women during the first six months after labor and were compared with 21 time-adjusted nutritional formulas. Animal experiments were performed on male Wistar rats. KYNA was administered in drinking water. The content of KYNA in human milk increases more than 13 times during the time of breastfeeding while its level is significantly lower in artificial formulas. KYNA was detected in breast milk of rats and it was found that the supplementation of rat maternal diet with KYNA in drinking water results in its increase in maternal milk. By means of the immunoblotting technique, GPR35 was evidenced in the mucosa of the jejunum of 1-day-old rats and distinct morphological changes in the jejunum of 21-day-old rats fed by mothers exposed to water supplemented with KYNA were found. A significant reduction of body weight gain of rats postnatally exposed to KYNA supplementation without changes in total body surface and bone mineral density was observed. The rat offspring fed with breast milk with artificially enhanced KYNA content demonstrated a lower mass gain during the first 21 days of life, which indicates that KYNA may act as an anti-obesogen. Further studies are, therefore, warranted to investigate the mechanisms regulating KYNA secretion via breast milk, as well as the influence of breast milk KYNA on mass gain. In the context of lifelong obesity observed worldwide in children fed artificially, our results imply that insufficient amount of KYNA in baby formulas could be considered as one of the factors associated with increased mass gain.

Maternal ß-hydroxy-ß-methylbutyrate (HMB) supplementation during pregnancy affects early folliculogenesis in the ovary of newborn piglets.

Beta-hydroxy-beta-methylbutyrate (HMB) is a leucine metabolite with protein anabolic effects. This study was designed to determine whether prenatal HMB treatment has an effect on oogenesis and folliculogenesis in the ovary of newborn piglets. HMB decreased the number of egg nests and primordial follicles and increased the pool of developing follicles compared to the control group. Although the percentage of TUNEL-positive oocytes within the egg nests was higher in HMB-treated group no increase in the Bax/Bcl-2 ratio and active caspase-3 expression was observed. Moreover, the granulosa cell proliferation index and StAR protein expression were higher in HMB-treated group. In contrast to the control group, the expression of E-cadherins was reduced after the HMB treatment. In addition, a significant increase in the serum level of gonadotropins and steroid hormones was detected in HMB-treated piglets. In conclusion, prenatal HMB treatment dysregulates hormonal homeostasis which impairs early folliculogenesis in piglets.

Maternal HMB treatment affects bone and hyaline cartilage development in their weaned piglets via the leptin/osteoprotegerin system.

It has been demonstrated in animal studies that prenatal administration of ß-hydroxy-ß-methylbutyrate (HMB, metabolite of leucine) influences general growth and mechanical endurance of long bones in newborn offspring in sex-dependent manner. The present experiment was conducted to evaluate the effect of HMB treatment of pregnant sows on bone development in offspring at weaning. From 70th day until the 90th day of gestation, sows received either a basal diet (n = 12) or the same diet supplemented with HMB (n = 12) at the dose of 0.2 g/kg of body weight/day. Femora obtained from six males and females in each group weaned at the age of 35 days were examined. Maternal HMB treatment significantly enhanced body weight and changed bone morphology increasing femur mechanical strength in both sexes. Maternal HMB supplementation also elevated bone micro- and macroelement concentrations and enhanced content of proteoglycans in articular cartilage. Based on the obtained results, it can be concluded that maternal HMB supplementation in the mid-gestation period significantly accelerated bone development in both sexes by upregulation of a multifactorial system including leptin and osteoprotegerin. However, the sex (irrespective of the HMB treatment) was the factor which influenced the collagen structure in cartilages and trabecular bone, as demonstrated both by the Picrosirius red staining and performed analysis of thermal stability of collagenous tissues. The structural differences in collagen between males and females were presumably related to a different collagen maturity. No studies conducted so far provided a detailed morphological analysis of bone, articular cartilage, growth plate and the activities of the somatotropic and pituitary-gonadal axes, as well as leptin/osteoprotegerin system in weaned offspring prenatally treated with HMB. This study showed also the relationship between the maternal HMB treatment and bone osteometric and mechanical traits, hormones, and growth and bone turnover markers such as leptin, osteoprotegerin and insulin-like growth factor-1.

Subsequent somatic axis and bone tissue metabolism responses to a low-zinc diet with or without phytase inclusion in broiler chickens.

Zinc is required for normal bone development and cartilage formation. The purpose of this study was to assess the effect of with adding organic Zn (alone or phytase inclusion) at the reduced dose to growing male Ross 308 chickens on somatic axis and bone tissue metabolism. 200 one-day old broilers were divided into the negative control group fed diet without Zn or phytase inclusion, positive control group receiving Zn in the 100% of daily recommended dose from ZnO, and two experimental groups fed diet introduced Zn in 25% of daily recommendation as a glycine chelate (Zn-Gly) with or without phytase inclusion (500 FTU·kg-1). Supplemental organic Zn increased bone Zn and Mg content, serum IGF-1, growth hormone and leptin concentration. Additional phytase inclusion increased body weight gain, blood plasma Ca, Fe, Zn and osteocalcin concentration and tibia ash percentage when compared to the Zn-deprived control. Bone geometry, yield and ultimate strengths were enhanced in both organic Zn supplemented groups, and the overall mechanical strength parameters of bone were better in these groups than in the positive control group supplemented with standard dose of inorganic Zn. Also marked improvements in the thickness of articular and the growth plate cartilages as well as real bone volume and thickness of metaphyseal trabeculae were achieved in all broilers fed Zn-supplemented diet irrespective of phytase inclusion, however, the highest cancellous bone mass and the best trabecular structure were noted after ZnO supplementation. In concludion, although dietary organic Zn given to growing broilers in 25% of daily recommended dose improved general bone properties and mechanical strength, the obtained results do not allow to unambiguously state that organic Zn supplementation at this level, even after phytase inclusion, is sufficient for proper bone development.