Anabolic steroids induced changes at the level of protein expression: Effects of prolonged administration of testosterone and nandrolone to pigs.

Synthetic derivatives of steroid hormones, specifically anabolic-androgenic steroids (AAS), have gained prominence due to their observed benefits in enhancing meat quality. The study replicated the administration of banned AAS and investigated their impacts on pigs to contribute to the understanding of animal biochemistry and to explore the feasibility of detecting AAS administration by employing a non-targeted analysis. The effects were corroborated by evaluating changes in the expression of selected proteins, as well as examining haematological and biochemical profiles and histological alterations. Exposure to AAS influenced the expression of proteins related to drug-metabolizing enzymes, muscle and lipid metabolism, kidney function, reproductive processes, immune system functions, and carcinogenic changes. The effects of AAS appear intricate and contingent on factors such as the specific drug used, dosage, and duration of administration. The results underscore that protein expression analysis holds promise as a valuable tool for detecting illicit AAS use in the fattening process.

Structural Changes in the Skeletal Muscle of Pigs after Long-Term Administration of Testosterone, Nandrolone and a Combination of the Two.

Anabolic steroid hormones (AASs) are used in most countries of the world to accelerate the growth of animals, increase the volume of their muscles and thereby increase meat production. However, there is a strict ban on the use of AASs in the fattening of all animals in all countries of the European Union, and there must therefore be effective methods of detection and control of these substances. Methods based on chromatography and mass spectrometry may no longer be completely effective when faced with new synthetic steroids of unknown chemical structures and low concentrations. Therefore, there is an effort to develop new methods of AAS detection, based primarily on the monitoring of biological changes at the level of gene expression or changes in metabolism or structure at the cellular level. More detailed knowledge of the mechanisms of action of AASs on tissues is essential for these methods, and histological changes are one of them. In this study, we report histological changes in muscle structure after AAS application, specifically in the size of muscle fibers, the amount of endomysium and the number of nuclei and satellite cells in muscle fibers. A pig model was also intentionally used for the study, as no such study has been carried out on this species, and at the same time, pork is one of the most consumed meats across Europe. The results of histology and fluorescent antibody labeling showed that AASs increased the diameter and surface area of muscle fibers and also significantly increased the number of satellite cells on the fiber surface. The evident correlations between the number of satellite cells, all nuclei and the diameters of muscle fibers between some experimental groups provide evidence that the selected histological parameters could be additional detection mechanisms for screening a large number of samples and indicate the possibility of the presence of AASs in pork meat in the future.

Effect of Polymeric Nanoparticles with Entrapped Fish Oil or Mupirocin on Skin Wound Healing Using a Porcine Model.

The utilization of poly(lactic-co-glycolic) acid (PLGA) nanoparticles (NPs) with entrapped fish oil (FO) loaded in collagen-based scaffolds for cutaneous wound healing using a porcine model is unique for the present study. Full-depth cutaneous excisions (5 × 5 cm) on the pig dorsa were treated with pure collagen scaffold (control, C), empty PLGA NPs (NP), FO, mupirocin (MUP), PLGA NPs with entrapped FO (NP/FO) and PLGA NPs with entrapped MUP (NP/MUP). The following markers were evaluated on days 0, 3, 7, 14 and 21 post-excision: collagen, hydroxyproline (HP), angiogenesis and expressions of the COX2, EGF, COL1A1, COL1A3, TGFB1, VEGFA, CCL5 and CCR5 genes. The hypothesis that NP/FO treatment is superior to FO alone and that it is comparable to NP/MUP was tested. NP/FO treatment increased HP in comparison with both FO alone and NP/MUP (day 14) but decreased (p < 0.05) angiogenesis in comparison with FO alone (day 3). NP/FO increased (p < 0.05) the expression of the CCR5 gene (day 3) and tended (p > 0.05) to increase the expressions of the EGF (day 7, day 14), TGFB1 (day 21) and CCL5 (day 7, day 21) genes as compared with NP/MUP. NP/FO can be suggested as a suitable alternative to NP/MUP in cutaneous wound treatment.

Importance of Zinc Nanoparticles for the Intestinal Microbiome of Weaned Piglets.

The scientific community is closely monitoring the replacement of antibiotics with doses of ZnO in weaned piglets. Since 2022, the use of zinc in medical doses has been banned in the European Union. Therefore, pig farmers are looking for other solutions. Some studies have suggested that zinc nanoparticles might replace ZnO for the prevention of diarrhea in weaning piglets. Like ZnO, zinc nanoparticles are effective against pathogenic microorganisms, e.g., Enterobacteriaceae family in vitro and in vivo. However, the effect on probiotic Lactobacillaceae appears to differ for ZnO and zinc nanoparticles. While ZnO increases their numbers, zinc nanoparticles act in the opposite way. These phenomena have been also confirmed by in vitro studies that reported a strong antimicrobial effect of zinc nanoparticles against Lactobacillales order. Contradictory evidence makes this topic still controversial, however. In addition, zinc nanoparticles vary in their morphology and properties based on the method of their synthesis. This makes it difficult to understand the effect of zinc nanoparticles on the intestinal microbiome. This review is aimed at clarifying many circumstances that may affect the action of nanoparticles on the weaning piglets' microbiome, including a comprehensive overview of the zinc nanoparticles in vitro effects on bacterial species occurring in the digestive tract of weaned piglets.

Effect of Milk Thistle (Silybum marianum) Seed Cakes by Horses Subjected to Physical Exertion.

Milk thistle (Silybum marianum) provides several possible benefits for horses, namely anti-inflammatory, antioxidant effects, hepatoprotective, and neuroprotective effects. Silybin exerts also pronounced effects on energy metabolism, that could be useful for sport horses. The aim of this study is to evaluate the effect of milk thistle seed cakes (in the form of a granulated mixture with barley) on blood biochemical parameters (total protein, albumin, bilirubin, urea, creatinine, ALT, AST, ALP, GGT, LDH, the total cholesterol, HDL-cholesterol, LDL-cholesterol, TAG, BHB, NEFA, creatine kinase, lactate, glucose, glutathione peroxidase, total antioxidant capacity, cortisol, calcium, and phosphorus) to monitor the differences between the experimental (milk thistle in feed) and control horses after exposure of the monitored horses to heavy physical exercise (combined driving) total number of horses was 12 Czech Warmblood breed horses. The digestibility of silymarin (and its individual flavonolignans) and basic nutrients are also monitored in this study. Statistically significant differences (P < .05) were found mainly in plasma cortisol and NEFA levels. The results of this study confirmed the results of our previous studies. These results suggest that the feeding of milk thistle seed cakes has a possible positive effect on horse health and energy metabolism.

Dose Effect of Milk Thistle (Silybum marianum) Seed Cakes on the Digestibility of Nutrients, Flavonolignans and the Individual Components of the Silymarin Complex in Horses.

Milk thistle seeds contain a mixture of flavonoids known as silymarin, which consists of silybin, isosilybin, silychristine, and silydianin. Until now, there has been no evidence of monitoring the digestibility of silymarin complex in horses. The aim of the research was to evaluate the digestibility of silymarin complex and the effect of nutrient digestibility in horses. Different daily feed doses (FD) of milk thistle expeller (0 g, 100 g, 200 g, 400 g, 700 g) were administered to five mares kept under the same conditions and at the same feed rations. Digestibility of silymarin complex was monitored by HPLC-UV. Digestible energy (DE), crude protein, crude fat, crude fiber, nitrogen-free extract (NFE), crude ash, calcium (Ca), and phosphorus (P) were determined according ISO/IEC 17025:2017. The biochemical profile of blood plasma (total protein, albumin, alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), bilirubin, total cholesterol, HDL cholesterol, LDL cholesterol, triacyl glyceride (TAG), non-esterified fatty acid (NEFA), creatine kinase (CK), creatinine, urea, glutathione peroxidase (GSH-Px), total antioxidant status (TAS), glucose, calcium, and inorganic phosphate) was investigated. Moreover, the flavonolignans of the silymarin complex in plasma were detected. Statistically significant differences (p ≤ 0.05) were found between daily doses of milk thistle expellers in digestibilities. Our findings showed the digestibility of flavonolignans increased with the daily dose and then stagnated with the dose of milk thistle seed cakes at 700 g/day.

Three-Dimensional Avian Hematopoietic Stem Cell Cultures as a Model for Studying Disease Pathogenesis.

Three-dimensional (3D) cell culture is attracting increasing attention today because it can mimic tissue environments and provide more realistic results than do conventional cell cultures. On the other hand, very little attention has been given to using 3D cell cultures in the field of avian cell biology. Although mimicking the bone marrow niche is a classic challenge of mammalian stem cell research, experiments have never been conducted in poultry on preparing in vitro the bone marrow niche. It is well known, however, that all diseases cause immunosuppression and target immune cells and their development. Hematopoietic stem cells (HSC) reside in the bone marrow and constitute a source for immune cells of lymphoid and myeloid origins. Disease prevention and control in poultry are facing new challenges, such as greater use of alternative breeding systems and expanding production of eggs and chicken meat in developing countries. Moreover, the COVID-19 pandemic will draw greater attention to the importance of disease management in poultry because poultry constitutes a rich source of zoonotic diseases. For these reasons, and because they will lead to a better understanding of disease pathogenesis, in vivo HSC niches for studying disease pathogenesis can be valuable tools for developing more effective disease prevention, diagnosis, and control. The main goal of this review is to summarize knowledge about avian hematopoietic cells, HSC niches, avian immunosuppressive diseases, and isolation of HSC, and the main part of the review is dedicated to using 3D cell cultures and their possible use for studying disease pathogenesis with practical examples. Therefore, this review can serve as a practical guide to support further preparation of 3D avian HSC niches to study the pathogenesis of avian diseases.