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Feed is the largest cost in pig production, driving interest in alternative ingredients that maintain performance at lower expense. Alfalfa, a nutrient-rich and widely available forage legume, shows promise as a partial feed substitute. However, it remains unclear how partial replacement with alfalfa affects growth dynamics, feed efficiency, and feeding behavior across fattening stages in pigs. This study investigated the effects of substitution with 8% alfalfa or alfalfa silage on growth performance and feeding behavior in growing-finishing pigs over a 60-day fattening period (60–130 kg). A total of 24 pigs were randomly assigned to three diet treatments: commercial feed, 8% alfalfa substitution, or 8% alfalfa silage substitution. Weight, feed intake, feed conversion ratio (FCR), and feeding hour were measured and analyzed using linear mixed-effects models and generalized additive mixed models. Results showed no significant effects of alfalfa on overall weight, feed intake or FCR. However, Pigs fed commercial feed showed superior FCR during mid-fattening (weeks 2.8–5.2, P < 0.05) but worse FCR than alfalfa-fed groups during late-finishing phase (weeks 6.6–8, P < 0.05). Moreover, alfalfa silage altered animal behavior by shifting feeding hour by roughly three hours earlier (P < 0.05). In summary, substitution with 8% alfalfa or alfalfa silage can be incorporated without compromising overall pig performance. While, integrating alfalfa in later fattening stages could further improve feed efficiency and reduce costs. This supports the use of phase-specific feeding strategies and adoption of such practices could enhance both economic and environmental sustainability in commercial pig farming.

Indigenous pigs constitute crucial genetic reservoirs. Adipose tissue is central to pig growth and metabolism, yet its molecular ontogeny remains poorly characterized in indigenous breeds such as the Ningxiang pig. We employed mass spectrometry to profile backfat proteomes across six postnatal stages (60-360 days). Proteomes clearly separated early (60-120 days) from late stages (300-360 days). Older pigs showed enrichment in processes linked to energy metabolism, translation, immune function, and mitochondrial activity. We identified 43 lipid metabolism proteins exhibiting significant age-dependent abundance. Weighted co-abundance network analysis revealed four protein modules significantly correlated with lean meat percentage, fat percentage, and carcass weight. Intramodular analysis identified four hub proteins-ALDH18A1, FABP4, FBP1, and HADHB-as putative candidates associated with lipid transport, gluconeogenesis, and fatty acid oxidation. This study links temporal proteomic profiles with key carcass traits, providing a data resource and a network-based framework for future research.

The aim of this study was to investigate whether supplementation with chitosan (COS) could reduce diarrhea and to explore how COS alleviates intestinal inflammation in weaned pigs. Thirty pigs (Duroc×Landrace×Yorkshire, initial BW of 5.65±0.27) weaned at age 21 d were challenged with enterotoxigenic Escherichia coli during a preliminary trial period, and then divided into three treatment groups. Pigs in individual pens were fed a corn-soybean meal diet, that contained either 0 (control), 50 mg/kg chlortetracycline, or 300 mg/kg COS for 21 days. The post-weaning diarrhea frequency, calprotectin levels and TLR4 protein expression were decreased (P<0.05) in both the COS and chlortetracycline groups compared with control. Simultaneously, supplemental COS and chlortetracycline had no effect on the mRNA expression of TNF-α in the jejunal mucosa, or on the concentrations of IL-1β, IL-6 and TNF-α in serum. However, COS supplementation improved (P<0.05) the mRNA expression of IL-1β and IL-6 in the jejunal mucosa. The results indicate that supplementation with COS at 300 mg/kg was effective for alleviating intestinal inflammation and enhancing the cell-mediated immune response. As feed additives, chitosan and chlortetracycline may influence different mechanisms for alleviating inflammation in piglets.

Understanding the metabolic profile of backfat is essential for optimizing breeding strategies and improving pork production. While the early stages of adipose tissue development have been partially characterized, metabolic alterations during the fattening phase (180-360 days)- a critical period for physiological maturation and carcass trait formation-remain insufficiently understood. In this study, we characterized the metabolomic profiles of backfat from Ningxiang pigs across four developmental stages (180d, 240d, 300d, 360d) and evaluated their relationships with carcass traits. A total of 154 metabolites exhibited significant temporal variation (q < 0.05), including functional lipids such as methanandamide phosphate and oleoylethanolamide, which are associated with appetite regulation and lipid metabolism. The progressive accumulation of bioactive exogenous metabolites like α-tocotrienol and cephaeline, indicated potential stage-dependent immunometabolic adaptations. In contrast, several synthetic xenobiotics, potentially derived from feed additives or environmental exposure, accumulated in backfat tissue and may represent potential risks to animal health and pork safety. Co-expression network analysis identified a metabolite module strongly associated (|R| > 0.7, q < 0.05) with key carcass traits, within which psychotrine-an understudied plant-derived alkaloid not previously associated with animal growth- was identified as a hub metabolite. This study establishes a comprehensive metabolic characterization of backfat during the fattening phase in an indigenous pig breed. Collectively, these findings provide novel candidate biomarkers for carcass trait prediction, highlight the potential impact of synthetic compound accumulation, and offer valuable insights for precision breeding, nutritional management, and meat safety assessment in swine production.

Differentiation of adipocytes and their aggregation to adipose tissue are critical for mammalian growth and development. MicroRNAs (miRNAs) are a class of endogenous small non-coding RNAs that play important roles in adipogenesis and lipid metabolism. miR-128-3p may contribute to adipose tissue development according to the previous studies. However, the role of miR-128-3p in the process of preadipocyte differentiation and lipid metabolism is not yet understood. The purpose of this research was to investigate the biological function and molecular mechanism of miR-128-3p in 3T3-L1 cells. In the present study, we found that miR-128-3p was downregulated during the process of 3T3-L1 preadipocyte differentiation. Overexpression of miR-128-3p obstructed the expressions of adipogenic marker genes as well as the lipid droplets accumulation and triglyceride content, suggesting the importance of miR-128-3p for adipogenesis. Moreover, miR-128-3p could lead to the retardation of cell proliferation in 3T3-L1 preadipocytes. Further evidences showed that, as a negative regulator of adipogenesis, miR-128-3p could directly target peroxisome proliferator-activated receptor γ ( Pparg ) which resulted in the suppression of 3T3-L1 preadipocyte differentiation, and miR-128-3p could also bind with SERTA domain containing 2 ( Sertad2 ) which drove triglyceride hydrolysis and lipolysis. In addition, inhibition of Sertad2 with siRNA displayed the same effects as overexpression of miR-128-3p. Our research demonstrated that miR-128-3p impeded 3T3-L1 adipogenesis by targeting Pparg and Sertad2 , resulting in the obstruction of preadipocyte differentiation and promotion of lipolysis. Taken together, this study offers profound insight into the mechanism of miRNA-mediated adipogenesis and lipid metabolism.

The aim of this study was to investigate whether supplementation with chitosan (COS) could reduce diarrhea and to explore how COS alleviates intestinal inflammation in weaned pigs. Thirty pigs (DurocLandraceYorkshire, initial BW of 5.65 plus or minus 0.27) weaned at age 21 d were challenged with enterotoxigenic Escherichia coli during a preliminary trial period, and then divided into three treatment groups. Pigs in individual pens were fed a corn-soybean meal diet, that contained either 0 (control), 50 mg/kg chlortetracycline, or 300 mg/kg COS for 21 days. The post-weaning diarrhea frequency, calprotectin levels and TLR4 protein expression were decreased (P<0.05) in both the COS and chlortetracycline groups compared with control. Simultaneously, supplemental COS and chlortetracycline had no effect on the mRNA expression of TNF- alpha in the jejunal mucosa, or on the concentrations of IL-1 beta , IL-6 and TNF- alpha in serum. However, COS supplementation improved (P<0.05) the mRNA expression of IL-1 beta and IL-6 in the jejunal mucosa. The results indicate that supplementation with COS at 300 mg/kg was effective for alleviating intestinal inflammation and enhancing the cell-mediated immune response. As feed additives, chitosan and chlortetracycline may influence different mechanisms for alleviating inflammation in piglets.

Nano-hydroxyapatite/bamboo fiber (n-HA/BF) composite membranes were obtained by a simple casting technology. The mechanism of the membrane formation and the effects of different pre-drying conditions, drying methods and n-HA addition amounts on the properties of the n-HA/BF composite membranes were investigated by Fourier Transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), Scanning electron microscopy (SEM), contact angle, Electromechanical universal tester, in vitro soaking in simulated body fluid (SBF) and in vitro cell culture experiment. The results demonstrated that different preparation conditions did not affect the dispersity of n-HA nanoparticles in BF matrix, but determined the membrane appearances, owing to the different changes of hydrogen bond under different pre-drying temperatures. Moreover, the hydrophilicity of the composite membranes could be improved when the membranes were prepared at 70 °C and freeze-drying as well as n-HA high addition content, and the mechanical properties of the composite membranes depended on n-HA addition content. In vitro soaking behavior indicated that the corrosion properties and bone-like apatite deposition could be controlled by different preparation conditions. The cell proliferation results showed that the n-HA/BF composite membranes obtained in different preparation conditions were all non-toxic. The obtained results showed that the casting technique could be used to prepare n-HA/BF composite membranes, and the properties of the composite membranes could be controlled by adopting different preparation conditions, which would have a great potential for guide bone tissue regeneration (GBR) membranes, and the study would provide a new way for BF application in biomedical fields. Graphical abstract In this manuscript, nano-hydroxyapatite/BF (n-HA/BF) composite membranes were prepared by casting method, and the membrane forming mechanism and the effects of different pre-drying conditions, drying methods and n-HA amounts on the corresponding n-HA/BF membrane were investigated. Results demonstrated that the morphologies of membrane was determined by the different preparation conditions owing to different hydrogen bond changes. Moreover, the hydrophilicity, the mechanical properties, the degradability and bone-like apatite deposition could be controlled by different preparation conditions, and all the n-HA/BF composite membranes were all non-toxic. The obtained results indicated that the n-HA/BF composite membrane with 20% n-HA prepared at room temperature has a great potential as guide bone tissue regeneration (GBR) membrane, which would provide a new application for BF in biomedical field.

Arbuscular mycorrhizal (AM) fungi can affect plant growth by regulating competition. Nutrient-deficient karst habitats contain abundant plants that compete for nutrients through interspecific or intraspecific competition, involving the nutritional transformation of litter decomposition. However, how plant competition in the presence of AM fungi and litter affects root development and nutrition remains unclear. A potted experiment was conducted, including AM fungus treatment with or without Glomus etunicatum , the competition treatment concerning intraspecific or interspecific competition through planting Broussonetia papyrifera and Carpinus pubescens seedlings, and the litter treatment with or without the mixture of B. papyrifera and C. pubescens litter leaves. The root morphological traits were analyzed, and nitrogen (N), phosphorus (P), and potassium (K) were measured. The results showed that AM fungus differently affected the root morphological development and nutrition of both competitive plants, significantly promoting B. papyrifera roots in the increase of dry weight, length, volume, surface area, tips, and branches as well as N, P, and K acquisitions regardless of litter addition. However, there was no apparent influence for C. pubescen s roots, except for the diameter in the interspecific competition with litter. The root dry weight, length, volume, surface area, and tips of B. papyrifera under two competitive styles were significantly greater than C. pubescen s regulated by AM fungus, presenting significant species differences. The responses of the relative competition intensity (RCI) on root morphological and nutritional traits indicated that AM fungus and litter both asymmetrically alleviated more competitive pressure for B. papyrifera than C. pubescen s, and the interspecific competition facilitated more root morphological development and nutrition utilization by endowing B. papyrifera root superiority relative to C. pubescens compared with the intraspecific competition. In conclusion, interspecific competition is more beneficial for plant root development and nutrition than intraspecific competition in the presence of AM fungus and litter via asymmetrically alleviating competitive pressure for different plants.