Age-Dependent Variations in Functional Quality and Proteomic Characteristics of Canine (Canis lupus familiaris) Epididymal Spermatozoa.

Increased male age is associated with a significant reduction in semen quality. Little is known about the sperm proteome changes resulting from the aging process. This study aimed to investigate the relationship between the functional quality and proteome of epididymal spermatozoa of dogs that were differing in age. The study was conducted on 30 male dogs that were divided into three age groups. G1-12 to 41 months old, G2-42 to 77 months old, and G3-78 to 132 months old. The sperm samples were assessed using a computer-assisted semen analysis (CASA). The epididymal sperm proteins were analyzed using gel electrophoresis (SDS-PAGE), nano-liquid chromatography coupled to quadrupole time of flight mass spectrometry (NanoUPLC-Q-TOF/MS) and bioinformatic tools. The sperm quality parameters were significantly lower in older dogs. NanoUPLC-Q-TOF/MS identification resulted in 865 proteins that were found in the G1, 472 in G2, and 435 in G3. There were seven proteins that were present in all three age groups, and four of them (ACTB, CE10, NPC2, CRISP2) showed significant changes among the studied groups. Age-dependent variations were detected in the sperm proteome composition and were related to important metabolite pathways, which might suggest that several proteins are implicated in sperm maturation and could be potential aging biomarkers.

Proteomic Analysis of Intracellular and Membrane-Associated Fractions of Canine (Canis lupus familiaris) Epididymal Spermatozoa and Sperm Structure Separation.

This study was provided for proteomic analysis of intracellular and membrane-associated fractions of canine (Canis lupus familiaris) epididymal spermatozoa and additionally to find optimal sonication parameters for the epididymal sperm morphological structure separation and sperm protein isolation. Sperm samples were collected from 15 dogs. Sperm protein fractions: intracellular (SIPs) and membrane-associated (SMAPs) were isolated. After sonication, sperm morphology was evaluated using Spermac Stain™. The sperm protein fractions were analyzed using gel electrophoresis (SDS-PAGE) and nanoliquid chromatography coupled to quadrupole time-of-flight mass spectrometry (NanoLC-Q-TOF/MS). UniProt database-supported identification resulted in 42 proteins identified in the SIPs and 153 proteins in the SMAPs. Differentially abundant proteins (DAPs) were found in SIPs and SMAPs. Based on a gene ontology analysis, the dominant molecular functions of SIPs were catalytic activity (50%) and binding (28%). Hydrolase activity (33%) and transferase activity (21%) functions were dominant for SMAPs. Bioinformatic analysis of SIPs and SMAPs showed their participation in important metabolic pathways in epididymal sperm, which may suggest their potential as sperm quality biomarkers. The use of sonication 150 W, 10 min, may be recommended for the separation of dog epididymal sperm heads, tails, acrosomes and the protein isolation.

The Effect of Diet Based on Legume Seeds and Rapeseed Meal on Pig Performance and Meat Quality.

The aim of this work was to assess the impact of a total dietary replacement of extracted soybean meal (SBM) on body weight gain and pork quality. DanBred hybrid piglets were divided into four groups of 10 piglets each. Groups I (males) and II (females) were the control groups and fed a standardized SBM-based complete feed. The experimental groups III (males) and IV (females) were offered a diet in which the SBM was replaced with extracted rapeseed meal (RSM) and legume plants (pea and yellow lupin). After 83 days of fattening, the animals were slaughtered. Based on the collected data, the daily weight gain (DWG), feed intake (FI), and feed conversion ratio (FCR) were calculated. In addition, longissimus dorsi muscle was subjected to physicochemical analyses, including the basic chemical composition. All the analyses were performed in accordance with the applicable methodologies. As a result of this experiment, no interactions were found between the experimental factors (sex and diet). The replacement of SMB by legumes and RSM resulted in a significant reduction in the final body weight of growing-finishing pigs. Additionally, daily body weight gain was reduced between 35-83 days, and through the whole fattening period (0-83 days). Most pork meat quality parameters were not affected by the type of mix feed and sex (p > 0.05). The fattening pigs that were fed legume seeds and RSM had significantly reduced fatness (p > 0.05) compared to the control pigs. In males, there was also a significantly lower pH45 (p < 0.05). It can be concluded that feeding pigs with diets containing legume plants and extracted rapeseed meal does not affect the pork meat quality, but it may worsen the body weight gain.

Growth performance and Carcass quality in broiler chickens fed on legume seeds and rapeseed meal.

The aim of this study was to compare the growth performance parameters, carcass quality, and meat traits in broiler chickens fed on diets containing legume seeds and rapeseed meal as an alternative to soybean meal. In this study, 448 male ROSS 308 chicks were divided into subgroups: a control group (I) fed on soybean meal (SBM), and six experimental groups II-rapeseed meal (RSM); III-white lupin (WY); IV-yellow lupin (YL); V-narrow-leaved lupin NLL; VI-pea (Pe); and VII-faba bean (FB). After 42 days of rearing, 10 birds from each group were slaughtered and dissected. The control group was characterized by better growth performance compared to the other groups. In addition, the European Broiler Index was lower in each experimental group compared to the SBM group. A lower dressing percentage was found only in the NLL group. The muscle content in birds from the RSM and FB groups was significantly higher than in the other groups, but the fat content was lower. Meat from SBM group was characterized by the highest protein content, but a reduced content of fat and water in the muscles. The most similar results were found between the control group and the FB group receiving a diet based on faba beans. Furthermore, lupins had a similar effect on the carcass traits when used in the diets. The quality of meat in broilers fed on faba beans with the addition of potato protein and brewers' yeast was similar to that of those fed on soybean meal, because the antinutrients were the lowest in faba bean seeds. Faba beans are proposed as a possible alternative source of protein in poultry diets. Other legume seeds should be analyzed in future studies.

Tacrolimus (FK506) and cyclosporin A reduce macrophage recruitment to the rat brain injured at perinatal and early postnatal periods.

OBJECTIVE: Tacrolimus (FK506) and cyclosporin A (CsA), immunosuppressants widely used in post-transplantional therapy, have been reported to protect neurons in the injured brain. This effect can be exerted directly and indirectly via inflammatory cells. Since the data come exclusively from studies on the adult brain, we examined effects of the drugs on the macrophage recruitment in the brain injured at early developmental stages. METHODS: Following the brain injury, 1- and 6-day-old Wistar rats (P1s and P6s, respectively) were treated with FK506 or CsA and injected with [(3)H]thymidine. Brain sections were processed for BSI-B4 isolectin histochemistry and subjected to autoradiography to visualize proliferating and non-proliferating macrophages. RESULTS: In P1s (n=33), FK506 evoked a dose-dependent reduction in the number of macrophages. P6s (n=30) presented greater decreases in macrophage numbers and their proliferative activity than the newborns. CsA application in P1s (n=27) affected neither recruitment of macrophages to the region of injury nor their proliferation. In CsA-treated P6s (n=28), reduction of the macrophage population and its proliferative activity was also seen but was much smaller than that following FK506 administration. DISCUSSION: High effectiveness of FK506 in regulation of the inflammatory response and neuroprotection observed in the adult brain can also be considered as a possible indirect determinant of neuronal survival following the brain injury at very early developmental stages.