Search Results (66)

Asthma is a chronic airway disease characterized by inflammation, bronchial hyperresponsiveness, and airflow limitation, highlighting the need for novel bronchodilator agents. Cryptotanshinone (CT), a bioactive diterpenoid derived from Salvia miltiorrhiza, exhibits anti-inflammatory and vasodilatory properties; however, its direct effects on airway smooth muscle remain poorly characterized. This study investigated the bronchodilatory activity of CT and its pharmacological mechanisms. Molecular docking was performed to evaluate potential interactions with M₃ muscarinic receptors and L-type calcium channels. Functional experiments were conducted using isolated guinea pig tracheal smooth muscle preparations. The relaxant effects of CT (10⁻⁷–3 × 10⁻⁴ M) were evaluated against carbachol (1 µM)- and high-K⁺ (80 mM)-induced contractions. Docking predicted favorable binding of CT to the M₃ receptor and L-type Ca²⁺ channel, with binding energies of −9.854 and −9.951 kcal/mol, respectively. In vitro, CT produced concentration-dependent relaxation of CCh-induced contractions, reaching a maximal effect of 41.9 ± 2.58% at 3 × 10⁻⁴ M (pEC₅₀ = 4.60). CT produced minimal relaxation in high-K⁺-induced contractions, suggesting receptor-mediated rather than non-selective smooth muscle inhibition. CT also produced a parallel rightward shift of the CCh concentration–response curve at 10⁻⁵ M, whereas a higher concentration (10⁻⁴ M) altered the maximal contractile response, suggesting concentration-dependent pharmacological effects. Pharmacological inhibition studies indicated the involvement of muscarinic receptor-mediated mechanisms, with additional contributions from calcium channel-related mechanisms and partial involvement of the NO/cGMP pathway, while β₂-adrenergic signaling and potassium channels were not significantly involved. These findings suggest that CT exerts bronchodilatory effects through the involvement of multiple pharmacological pathways relevant to airway smooth muscle regulation and provide preliminary mechanistic evidence supporting further investigation. Full article

Shuanghuanglian oral liquid (SHL) is widely used in companion animals and poultry, but its molecular mechanism in pneumonia–myocarditis comorbidity and heart–lung inflammatory crosstalk remains largely unclear. This computational study investigated the conserved AKT1/HIF1A-mediated immunoregulatory mechanism of SHL and its cross-species translational potential in veterinary medicine. Network pharmacology was integrated with GO, KEGG, and Reactome enrichment analyses, protein–protein interaction network construction, ADMET evaluation, cross-species sequence homology analysis (human, dog, cattle, and pig), molecular docking, and molecular dynamics simulation. A total of 61 active compounds, 251 putative targets, and 52 common targets associated with pneumonia and myocarditis were identified. These targets were mainly enriched in inflammation- and immune-related pathways, including TNF, IL-17, AGE–RAGE, and PPAR signaling. AKT1 and HIF1A showed high sequence conservation across species (85–98%). Key compounds exhibited favorable binding affinity to AKT1, and molecular dynamics simulation suggested the stability of the Baicalein–AKT1 complex. ADMET analysis suggested favorable pharmacokinetic properties and low predicted toxicity. These findings suggest that SHL may potentially alleviate pneumonia and myocarditis through modulation of the conserved AKT1/HIF1A axis and support its potential as a complementary therapeutic approach for managing heart–lung inflammatory diseases in multiple livestock species. This entirely computational study highlights promising mechanisms that should be further validated in vivo. Full article

Animal welfare monitoring is essential in pig production. On-farm animal welfare (AW) assessments may provide a comprehensive overview but are resource-intensive. Abattoir-based assessments allow pigs from multiple farms to be inspected in a single facility. However, data on the relationship between these assessments remain limited, especially for heavy pigs (160–170 kg). This study investigates these associations in Italian heavy pig production. At the abattoir, 18,333 pig carcasses from 185 batches across 86 farms were scored for tail, skin (cranial and caudal) and ear lesions. On-farm AW assessments (management, structures and animal-based measures) were obtained from the national surveillance system (ClassyFarm). Tail lesion scores were higher in pigs with intact tails, whereas ear scores showed the opposite trend, suggesting a substitution effect between tail and ear biting. This indicates that tail docking is insufficient to fully prevent abnormal behaviours. Higher skin and ear scores were associated with suboptimal management, but tail scores were not, likely due to the multifactorial nature of tail biting. Herd size had no significant effect on welfare indicators. These results highlight the complexity of assessing AW and the importance of combining abattoir and farm data to obtain a more integrated monitoring system. Full article

The porcine epidemic diarrhea virus (PEDV) causes a gastrointestinal disease generating mortality rates approaching 100% in piglets worldwide. The S glycoprotein of PEDV is the main target for the development of vaccines. Two vaccines approved by the Ministry of Agriculture and Rural Development are used in Mexico: the first vaccine is based on an inactivated virus isolated more than a decade ago, whereas the second vaccine is based on mRNA technology. The most important tool for controlling PEDV outbreaks is vaccination; however, coronaviruses are characterized by the accumulation of multiple mutations, which compromise the immune response elicited by outdated vaccines. In this work, we classified the Mexican strains of PEDV reported so far in GenBank, according to their genotypes. Subsequently, we searched for B and T cell epitopes conserved in Mexican PEDV strains using bioinformatic tools. In addition, we explored whether these epitopes can induce allergies, autoimmunity, and/or toxic effects. Next, we determined the localization of B cell epitopes in the S glycoprotein using the protein crystal and protein modeling of several S glycoproteins. Finally, we carried out molecular docking analysis to assess whether these T cell epitopes could interact with the peptide-binding groove of the Swine Leukocyte Antigens (SLAs). Five conserved B cell epitopes were found to be exposed on the surface of the S glycoprotein, whereas several promiscuous CTL and HTL epitopes were bound, with low free energy, to the peptide-binding grooves of SLA-I and SLA-II, respectively. The best epitopes were used to generate a plasmid carrying the sequence to produce a recombinant protein. This plasmid was used for transfection experiments in PK-15 cell culture. The B cell epitopes reported here were recognized by the sera from pigs infected with PEDV but not by the sera from uninfected animals. These results justify future evaluations of the ability of these epitopes to stimulate cytokine production by T cells, antibody generation, and their neutralizing activity. Full article

Toll-like receptor 7 (TLR7) and Stimulator of Interferon Genes (STING) ligands possess a series of immunomodulatory effects such as anti-infection, anti-tumor, and autoimmune-disease-alleviating effects. In this study, porcine TLR7 (pTLR7) and porcine STING (pSTING) were selected as targets, and molecular docking and virtual screening methods were used for screening of dual-target livestock immunomodulators. Finally, two compounds were screened with molecular docking scores higher than the positive control compounds. They have good binding ability with pTLR7 and pSTING proteins, as well as satisfactory predictive safety and pharmacokinetic properties. Molecular dynamics (MD) simulation results also indicated that the above ligands can form stable complexes with two target proteins. The average binding free energies of compound 2 with pTLR7 and pSTING were −28.65 kcal/mol and −30.12 kcal/mol, respectively, and of compound 7 with pTLR7 and pSTING were −35.93 kcal/mol and −31.70 kcal/mol, respectively, which were comparable to that of positive control ligands. The similarity of target proteins between pigs, humans, and mice, as well as the interactions between ligands and TLR7 and STING in different species, were analyzed. And analysis of predicted structure–activity relationship (SAR) was conducted. Briefly, compound 2 and compound 7 were predicted to form stable complexes with pTLR7 and pSTING, with satisfactory predicted physicochemical properties and pharmacokinetic characteristics, and represented candidates for experimental validation. This study supplies a research basis for the development, design, and structural modification of immune enhancers for animals. Full article

Background/Objectives: Streptococcus suis is a zoonotic pathogen that causes infections in pigs and humans, leading to significant economic losses. S. suis can evade the immune system of hosts and induce persistent infections. Early detection and vaccination are crucial for controlling the disease in swine industries. This study aimed to investigate candidate recombinant protein for antibodies against S. suis detection and subunit vaccine development. Methods: The whole genome of S. suis BM407 was analyzed using bioinformatic tools to predict suitable proteins and genes for recombinant protein expression. Partial extracellular factor protein (epf) genes of S. suis serotype 2 DMST18783 were amplified. A 3301 bp amplicon was digested, and a specific 615 bp fragment was inserted into a pQE81L-KAN vector. Then, the constructed plasmid was cloned and expressed in Escherichia coli DH10β. Purified protein was analyzed using SDS-PAGE. In addition, translated amino acid sequences were analyzed for immune response properties, molecular docking, molecular dynamic simulation, and epitope prediction. Results: The amino acid sequence of recombinant extracellular factor protein (rEF) was revealed as a promising antigen containing putative protective regions as linear epitopes. Furthermore, the rEF was expressed as a histidine-tagged recombinant protein, and its properties were nearly similar to the predicted rEF using bioinformatic tools. Binding of the recombinant EF (rEF) protein was found to reduce fluctuations in the swine toll-like receptor 2. Furthermore, the rEF contained several regions that were predicted to be epitopes for both B-cells and T-cells. Conclusions: This study indicates that the recombinant EF fragment is a promising candidate for detecting antibodies against S. suis and as a component of a subunit vaccine. Full article

This study aimed to evaluate the effects of no tail docking on the performance, health, and behavior of piglets raised under commercial conditions in Brazil. The study included 768 weaned piglets from the Pietrain synthetic line, randomly divided into two groups: DT = the final third part of the tail-docked (n = 384) and NTD = non-tail-docked (n = 384). Tail docking was performed on day two using an electrocautery clipper for piglets from the DT group, and both groups were subjected to standard environmental enrichment with branched chains. In cases of tail biting, a contingency plan was adopted to mitigate this problem by enriching the pen with a sisal rope. Behavioral measurements were performed using scan sampling. Tail biting, reactivity to humans, and health were assessed using a methodology adapted from the Welfare Quality Protocol^®. The piglets were weighed at 140 days of age and inspected according to the parameters established by the Pig Genealogical Registration Service to be used as reproduction animals. The off-test rate was calculated based on the total number of piglets approved for animal use relative to the total number evaluated. During the nursery stage, the NDT piglets showed a trend toward significance (p = 0.07) toward a higher occurrence of tail biting than the DT piglets and exhibited a higher incidence of severe lesions. They also engaged more frequently (p < 0.05) in exploratory behavior, interacting with branched chains and sisal rope, than the DT piglets. During the finishing phase, tail biting was observed only in the NDT piglets (p = 0.001). The NDT piglets that did not require the contingency plan exhibited lower fear responses (p = 0.02) during human interactions in the nursery phase than the DT piglets. Conversely, the NDT piglets that required a contingency plan showed higher fear levels (p < 0.001). Productivity performance was not affected (p > 0.05), and new cases of tail biting ceased after the contingency plan was implemented. The number of animals that died or were removed did not differ between the treatments (p > 0.05). In conclusion, managing piglets with intact tails on commercial farms presents a significant welfare challenge. By contrast, docking the final third of the tail, in accordance with regulations, was associated with fewer negative welfare outcomes, even when best management practices were applied. Full article

β-glucuronidase is an important hydrolase, which plays an important role in drug metabolism, clinical diagnostics, and biotransformation. This study focuses on the heterologous expression, isolation, purification, and its enzymatic properties of β-glucuronidase CpGUS from Clostridium perfringens, as well as its application in the whole-cell transformation of unconjugated bilirubin from pig bile. A recombinant E. coli BL21(DE3)/pET-28a-CpGUS was constructed for the heterologous expression of CpGUS, with the majority of the expressed enzyme being soluble. Enzymatic analysis showed that CpGUS displayed optimal activity at pH 5.0 and 45 °C, and it rapidly lost activity at pH < 4.5. Metal ions, such as Mg²⁺ and Fe²⁺, enhanced CpGUS catalysis, while Zn²⁺, K⁺, Fe³⁺, Mn²⁺, Cu²⁺, and Na⁺ inhibited it. Notably, Cu²⁺ and Fe³⁺ can significantly inhibit β-glucuronidase, resulting in the complete loss of its activity. The results of the whole-cell transformation experiment show that when E.coli BL21(DE3)/ pET-28a-CpGUS at an OD₆₀₀ of 10 was incubated at pH 5.0, a temperature of 45 °C, and a rotation speed of 200 rpm for 12 h, the hydrolysis rate of the conjugated bilirubin in pig bile reached 81.1%, the yield of unconjugated bilirubin was 76.8%, and the purity of unconjugated bilirubin was 98.2%. The three-dimensional structure of CpGUS was predicted using AlphaFold2 (AlphaFold v2.0, DeepMind Technologise Limited, London, UK), and p-Nitrophenyl-β-D-Glucuronide (pNPG) and conjugated bilirubin were then docked to the CpGUS protein model using SWISSDOCK. The best docked conformations of the CpGUS–pNPG and CpGUS–conjugated bilirubin complex systems were simulated by independent 500 ns molecular dynamics (MD) runs with the RSFF2C force field, and the binding dynamic and catalytic mechanism of each system were obtained. The results indicated that π-π stacking, hydrogen bonding, and hydrophobic interactions between the key residue Tyr472 and the benzene ring of pNPG molecules are crucial for its catalytic process. Similarly, for the binding and catalysis of conjugated bilirubin by CpGUS, the π-π stacking and hydrogen bonding and hydrophobic interactions between the sidechains of residues Phe368 and Tyr472 and the benzene ring of conjugated bilirubin play a synergistic role during its catalytic process. Their total binding free energy (∆G_bind) values were calculated to be as high as −65.05 ± 12.66 and −86.70 ± 17.18 kJ/mol, respectively. These results suggest that CpGUS possesses high binding and catalytic hydrolysis properties for both pNPG and conjugated bilirubin. Full article

The European Commission’s ban on routine tail docking has prompted this retrospective observational study to evaluate the short-term effects of transitioning to a fully undocked system. Twenty-two farms were assessed during three subsequent phases: total tail docking (step 1), subgroups of undocked pigs (step 2), and fully undocked pigs (step 3). Farmers received training in long-tail management and independently implemented it on their own farms. However, straw provision as environmental enrichment was mandatory, at least supplied during periods of pigs’ restlessness. Overall, going through step 2 appears to be successful. However, transitioning to step 3 worsened mortality (p = 0.010) and the feed conversion ratio (p = 0.015) in weaners. Compared to step 1, the cost of producing 1 kg of meat in step 3 was 33.9% greater during weaning and 7.4% during fattening. Tail lesion prevalence at slaughter was greater in step 3 (41%), followed by step 2 (10%) and step 1 (1%). The hypothetical labour required to optimize straw management compared to the adopted system, ensuring its continuous availability, was estimated as 35 min/100 piglets/weaning cycle (EUR 4.37) and 10.5 h/100 pigs/fattening cycle (EUR 109). Under the conditions of this study, transitioning to a fully undocked system was not successful. Mandating only the non-continuous use of straw has proven insufficient, and greater efforts must be systematically implemented. Full article

The objective of this research was to investigate the efficacy of lotus leaf in the prevention and treatment of inflammatory diarrhea in pigs, utilizing network pharmacology and in vitro methodologies. Initially, LC-MS was employed to analyze the constituents of lotus leaf extract (LLE); then, the TCMSP database was utilized to identify the active components and their corresponding targets. The GeneCards database was consulted to identify disease-related targets pertinent to inflammatory diarrhea in pigs. A drug ingredient–disease target network was constructed using Cytoscape software. Subsequently, the STRING database facilitated protein interaction analysis, which was also visualized through Cytoscape. Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were conducted based on the genes shared between disease and LLE targets. Molecular docking of the active ingredients with key targets was performed using Autodock Vina. Subsequently, an in vitro LPS-induced inflammation model was established using IPEC-J2 cells to validate the predictions made through network pharmacology. Verification was conducted via flow cytometry and Western blot analysis. The LC-MS assay and TCMSP retrieval results revealed that Quercetin, Nuciferine, Kaempferol, Leucodelphinidin, and Catechin were identified as the main compounds of LLE, associated with 181 potential targets. A total of 5995 targets were linked to inflammatory diarrhea in pigs, with 159 overlapping targets identified between the bioactive compounds and the disease. Notable key targets included TNF-α, IL-6, caspase-3, TP53, and AKT, which are integral to inflammation and apoptosis processes. GO functional annotation indicated significant enrichment in biological processes such as gene expression regulation and transcription from RNA polymerase II promoters. KEGG pathway analysis highlighted critical pathways, including TNF signaling and apoptosis. Furthermore, molecular docking analyses demonstrated that the bioactive components of lotus leaf exhibited a strong binding affinity for essential targets, including AKT1, BAX, caspase-3, CCL2, IL-6, IL-10, MPK14, NOS3, PTGS1, and TNF-α. In vitro experiments confirmed that LLE significantly inhibited LPS-induced apoptosis in IPEC-J2 cells and suppressed the activation of the TNF-α-mediated apoptosis pathway. This study offers novel insights into the therapeutic potential of Chinese medicine and its constituents in addressing inflammatory diarrhea in pigs. Full article

Lysophosphatidic acid (LPA) is a small bioactive phospholipid which plays an important role during embryonic development and promotes developmental potential of in-vitro-produced (IVP) embryos in several species, including sheep and pigs. In bovines, LPA accelerates IVP blastocyst formation through the Hippo/YAP pathway. However, other LPA effects and its potential receptors during bovine embryo development are less clear. In this study, we used enzyme-linked immunosorbent assay (ELISA) to assess the presence of LPA in bovine oviductal fluid and determine cell apoptosis in embryos after LPA stimulation by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and quantitative reverse transcription polymerase chain reaction (qRT-PCR). We further evaluated potential receptors of LPA through molecular docking, RNA-seq data analysis and quantitative RT-PCR. LPA was found to be present in oviductal fluid. An increase in total cell number and a decrease in apoptosis levels were detected in day 7 blastocysts after LPA treatment. Among eight LPA receptors (LPARs), GPR87 and LPAR2 showed the highest affinity with LPA and their transcripts were expressed in embryos after the 16-cell stage in RNA-seq and qRT-PCR analysis. However, only the expression of LPAR2 was significantly increased in day 6 blastocysts after LPA stimulation, indicating its potential role in LPA-mediated signaling pathways. Our data highlight the positive effects of LPA on embryos and enrich information of related signaling mediators of LPA during embryonic development. Full article

Tail docking is still used in pigs to reduce the prevalence of tail biting, although it is purely symptomatic and contrary to animal welfare. Genetic selection for shorter tails might, however, help to avoid tail docking and has therefore been proposed. A genetic basis for tail length is known for many species. Variability in tail length, including moderate heritability, has also been demonstrated in pigs. The aim of the present study was to identify genetic markers for tail length and to define candidate genes. To this end, 140 piglets were phenotyped and genotyped at 3 days of age and a genome-wide association study was performed. Seven SNPs were mapped on chromosomes 1, 2, 6, 11, and 15. Two linked SNPs on chromosome 2 resulted in a functional amino acid exchange. The genotypes at the SNPs were only associated with small differences in relative tail length of up to 16.5% (short genotype versus long genotype at SSC15), but at the same time with the occurrence of malformations in the form of tail kinks. The small effect size and the association between tail length and tail kinks, together with the generally pure symptomatic effect on tail biting, argue against the applicability of selection for shorter tails in pigs. Full article

Streptococcus suis (S. suis) is a major swine pathogen throughout the world as well as an emerging zoonotic agent. Among the symptoms caused by S. suis, including septicemia, pneumonia, endo-carditis, arthritis, and meningitis, the latter is the most overlooked. In the present study, we explored the mechanism of action of berberine against S. suis meningitis by obtaining berberine-related action targets, porcine S. suis meningitis targets, and human S. suis meningitis targets from open databases. We constructed a protein–protein interaction (PPI) network by using the STRING database and employed Cytoscape 3.8.0 to screen for core targets. We performed Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses through DAVID. We identified 31 potential targets of berberine, of which Toll-like receptor 4 (TLR4), fibronectin 1 (FN1), superoxide dismutase (SOD1), and catalase (CAT) were the four most critical targets. GO analysis revealed the enrichment of terms related to the response to oxidative stress and the inflammatory response. KEGG analysis revealed the enrichment of the interleukin 17 (IL-17), phosphoinositide 3-kinase (PI3K)-Akt, TLR, tumor necrosis factor (TNF), and mitogen-activated protein kinase (MAPK) signaling pathways. In addition, the admetSAR results showed that berberine can cross the blood–brain barrier. The molecular docking results indicated key binding activity between TLR4–berberine and FN1–berberine. In summary, berberine protects against Streptococcus suis meningitis by regulating inflammatory response and oxidative stress in humans and pigs. Our study updates the current knowledge of the targets of S. suis meningitis to exploit new drugs in humans and pigs, to develop environmentally friendly and antibiotic-free animal-derived food products, and to improve the farming industry and economic development. Full article

As a zoonotic pathogen, S. suis serotype 2 (SS2) can cause severe diseases in both pigs and humans, and develop resistance to antibiotics. Plant natural compounds are regarded as promising alternatives to conventional antibiotics. Phillyrin is the major bioactive components of Chinese herbal medicine Forsythia suspensa. In this study, we explored the activity and action mechanism of phillyrin against SS2. The results showed that phillyrin could disrupt membrane integrity, destroy intracellular structures, and increase the exosmosis of DNA. Results of PCR revealed that phillyrin affected bacterial-virulence-related genes’ expression levels. Meanwhile, phillyrin significantly decreased the adhesion activity, inhibited lactate dehydrogenase (LDH) secretion, and reduced biofilm formation of SS2 in Newborn pig trachea epithelial (NPTr) cells. Furthermore, phillyrin protected tight junction protein of NPTr cells from SS2. We reported that phillyrin (0.1 mg/kg) treatment after bacterial challenge significantly improved the survival rate, ameliorated pulmonary inflammation, and inhibited the accumulation of multiple cytokines (IL-1, IL-6, IL-8, and TNF-α). Molecular docking showed that phillyrin had a good binding activity with the Ala88 and Asp111 of suilysin (SLY), one of the most important virulence factors of SS2. Collectively, phillyrin possesses antibacterial and anti-inflammatory activities, and is a promising candidate for preventing SS2 infection. Full article

Fungal infections present a significant health risk, particularly in immunocompromised individuals. Berberine, a natural isoquinoline alkaloid, has demonstrated broad-spectrum antimicrobial activity, though its antifungal potential and underlying mechanisms against both yeast-like and filamentous fungi are not fully understood. This study investigates the antifungal efficacy of berberine against Candida albicans, Cryptococcus neoformans, Trichophyton rubrum, and Trichophyton mentagrophytes in vitro, as well as its therapeutic potential in a murine model of cryptococcal infection. Berberine showed strong antifungal activity, with MIC values ranging from 64 to 128 µg/mL. SEM and TEM analyses revealed that berberine induced notable disruptions to the cell wall and membrane in C. neoformans. No signs of cell necrosis or apoptosis were observed in fungal cells treated with 2 × MIC berberine, and it did not increase intracellular ROS levels or affect mitochondrial membrane potential. Molecular docking and binding affinity assays demonstrated a strong interaction between berberine and the fungal enzyme CYP51, with a dissociation constant (KD) of less than 1 × 10⁻¹² M, suggesting potent inhibition of ergosterol biosynthesis. In vivo studies further showed that berberine promoted healing in guinea pigs infected with T. mentagrophytes, and in a murine cryptococcal infection model, it prolonged survival and reduced lung inflammation, showing comparable efficacy to fluconazole. These findings indicate that berberine exerts broad-spectrum antifungal effects through membrane disruption and CYP51 inhibition, highlighting its potential as a promising therapeutic option for fungal infections. Full article