ABSTRACT
Enteric microbial pathogens, including Escherichia coli, Shigella and Cryptosporidium species, take a particularly heavy toll in low-income countries and are highly associated with infant mortality. We describe here a means to display anti-infective agents on the surface of a probiotic bacterium. Because of their stability and versatility, VHHs, the variable domains of camelid heavy-chain-only antibodies, have potential as components of novel agents to treat or prevent enteric infectious disease. We isolated and characterized VHHs targeting several enteropathogenic E. coli (EPEC) virulence factors: flagellin (Fla), which is required for bacterial motility and promotes colonization; both intimin and the translocated intimin receptor (Tir), which together play key roles in attachment to enterocytes; and E. coli secreted protein A (EspA), an essential component of the type III secretion system (T3SS) that is required for virulence. Several VHHs that recognize Fla, intimin, or Tir blocked function in vitro. The probiotic strain E. coli Nissle 1917 (EcN) produces on the bacterial surface curli fibers, which are the major proteinaceous component of E. coli biofilms. A subset of Fla-, intimin-, or Tir-binding VHHs, as well as VHHs that recognize either a T3SS of another important bacterial pathogen (Shigella flexneri), a soluble bacterial toxin (Shiga toxin or Clostridioides difficile toxin TcdA), or a major surface antigen of an important eukaryotic pathogen (Cryptosporidium parvum) were fused to CsgA, the major curli fiber subunit. Scanning electron micrographs indicated CsgA-VHH fusions were assembled into curli fibers on the EcN surface, and Congo Red binding indicated that these recombinant curli fibers were produced at high levels. Ectopic production of these VHHs conferred on EcN the cognate binding activity and, in the case of anti-Shiga toxin, was neutralizing. Taken together, these results demonstrate the potential of the curli-based pathogen sequestration strategy described herein and contribute to the development of novel VHH-based gut therapeutics.
Subject(s)
Bacterial Toxins , Cryptosporidiosis , Cryptosporidium , Enteropathogenic Escherichia coli , Probiotics , Single-Domain Antibodies , Humans , Antigens, Surface , Congo Red , Flagellin , Type III Secretion Systems , Virulence Factors/geneticsABSTRACT
Metal-organic gels (MOGs) are unique supramolecular gels that are convenient to synthesize. In this work, a cathodic electrochemiluminescence (ECL) system based on Ag-MOGs as a luminophore and K2S2O8 as a co-reactor was developed. The ECL spectrum of the Ag-MOGs overlapped significantly with the strong UV-Vis spectrum of the SiO2@PANI@AuNPs, which effectively quenched the ECL luminescence of the Ag-MOGs. Relying on the inner filter effect between Ag-MOGs and SiO2@PANI@AuNPs, a novel ECL-IFE immunosensor was developed for the detection of neuron-specific enolase (NSE). Under optimal conditions, the ECL signal of the immunosensor displayed excellent linearity over the NSE concentration range of 10 fg/mL-100 ng/mL. The limit of detection (LOD) was 2.6 fg/mL (S/N = 3) with a correlation coefficient R2 of 0.9975. The ECL immunosensor also exhibited excellent stability and reproducibility for the detection of NSE. The results reported provide a feasible concept for the development analytical methods for the detection of other clinically relevant biomarkers.
Subject(s)
Biosensing Techniques , Metal Nanoparticles , Gold , Silicon Dioxide , Luminescent Measurements/methods , Immunoassay/methods , Biosensing Techniques/methods , Reproducibility of Results , Electrochemical Techniques/methods , Gels , Phosphopyruvate HydrataseABSTRACT
BACKGROUND: Spinocerebellar ataxia type 3 (SCA3) is the most common autosomal dominant hereditary ataxia worldwide, which is however in a lack of effective treatment. In view of that engineered exosomes are a promising non-invasive gene therapy transporter that can overcome the traditional problem of poor drug delivery, the aim of this study was to evaluate, for the first time, the value of exosome-based microRNA therapy in SCA3 and the therapeutic effects of intravenously administrated ATXN3 targeting microRNAs in transgenic SCA3 mouse models. METHODS: The rabies virus glycoprotein (RVG) peptide-modified exosomes loaded with miR-25 or miR-181a were peripherally injected to enable targeted delivery of miRNAs to the brain of SCA3 mice. The behaviors, ATXN3 level, purkinje cell and other neuronal loss, and neuroinflammation were evaluated 4 weeks after initial treatment. RESULTS: The targeted and efficient delivery of miR-25 and miR-181a by modified exosomes substantially inhibited the mutant ATXN3 expression, reduced neuron apoptosis and induced motor improvements in SCA3 mouse models without increasing the neuroinflammatory response. CONCLUSIONS: Our study confirmed the therapeutic potential of engineered exosome-based miR-25 and miR-181a treatment in substantially reducing ATXN3 aggregation and cytotoxicity by relying on its targeted and efficient drug delivery performance in SCA3 mice. This treatment method shows a promising prospect for future clinical applications in SCA3.
Subject(s)
Exosomes , Machado-Joseph Disease , MicroRNAs , Mice , Animals , Machado-Joseph Disease/genetics , Machado-Joseph Disease/therapy , Mice, Transgenic , Apoptosis , MicroRNAs/geneticsABSTRACT
This study aimed to design a sandwich electrochemiluminescence (ECL) immunosensor with double co-reaction accelerators for sensitively detecting squamous cell carcinoma antigen (SCCA). First, silver orthophosphate (Ag3PO4) nanoparticles were modified on the surface of EuPO4 nanowires to improve their poor dispersibility/solubility. At the same time, EuPO4 was used as a co-reaction accelerator to catalyze S2O82- to produce more intermediates (SO4â¢-), significantly enhancing the ECL signal of Ag3PO4. Ag nanoparticles (AgNP) modified on Ag3PO4@EuPO4 composite nanomaterials were used not only as linkers of luminescence groups and biomarkers but also as a co-reaction accelerator to effectively enhance ECL signal. The designed ECL immunosensor displayed several advantages, including good stability and reproducibility. Under the optimal conditions, its linear range in detecting SCCA was 0.0001-50 ng·mL-1, the detection limit was 25 fg·mL-1 (S/N = 3), the recovery was 96.6-100.4%, and the relative standard deviation was less than 4.8%. It was successfully applied to detect SCCA in human serum.
Subject(s)
Antigens, Neoplasm , Biosensing Techniques , Metal Nanoparticles , Serpins , Humans , Electrochemical Techniques , Immunoassay , Luminescent Measurements , Reproducibility of Results , Silver , Antigens, Neoplasm/analysis , Serpins/analysisABSTRACT
Parkinson's disease (PD) is a neurodegenerative disease characterized by progressive death of dopaminergic neurons in the substantia nigra and the formation of Lewy bodies (LBs). Mutations in PD-related genes lead to neuronal pathogenesis through various mechanisms, with known examples including SNCA/α-synuclein (PAKR1), Parkin (PARK2), PINK1 (PARK6), DJ-1 (PARK7), and LRRK2 (PARK8). Molecular chaperones/co-chaperones are proteins that aid the folding of other proteins into a functionally active conformation. It has been demonstrated that chaperones/co-chaperones interact with PD-related proteins and regulate their function in PD. HSP70, HSP90 and small heat shock proteins can prevent neurodegeneration by regulating α-syn misfolding, oligomerization and aggregation. The function of chaperones is regulated by co-chaperones such as HSP110, HSP40, HOP, CHIP, and BAG family proteins. Parkin, PINK1 and DJ-1 are PD-related proteins which are associated with mitochondrial function. Molecular chaperones regulate mitochondrial function and protein homeostasis by interacting with these PD-related proteins. This review discusses critical molecular chaperones/co-chaperones and PD-related proteins which contribute to the pathogenesis of PD, hoping to provide new molecular targets for therapeutic interventions to thwart the disease progression instead of only bringing symptomatic relief. Moreover, appreciating the critical role of chaperones in PD can also help us screen efficient biomarkers to identify PD at an early stage.
Subject(s)
Dopaminergic Neurons/metabolism , Molecular Chaperones/metabolism , Parkinson Disease/metabolism , Substantia Nigra/metabolism , Ubiquitin-Protein Ligases/metabolism , alpha-Synuclein/metabolism , Animals , Disease Models, Animal , Dopaminergic Neurons/pathology , Humans , Parkinson Disease/pathology , Substantia Nigra/pathologyABSTRACT
BACKGROUND: Chitosan was used as an alternative to promote the growth of weaned piglets. And low-molecular-weight chitosan (LC) is one of chitosan derivatives and maintain beneficial biological properties of chitoson. The present experiment was carried out to examine the effects of LC on the growth performance, intestinal morphology, barrier function, cytokine expression, and antioxidant system of weaned piglets. RESULTS: A total of 40 piglets weaned at 21 d of age, with average body weight 6.37 ± 0.08 kg, were randomly assigned (5 pens/diet; 4 pigs/pen) to 2 treatments (a basal diet and the basal diet supplemented with 50 mg/kg LC) and were fed for 28 d. Compared with the control group, average daily feed intake (ADFI), and the expression of intestinal barrier protein ZO-1 was increased (P < 0.05) when the piglets fed the diet supplemented with LC. No significant differences were found in average daily gain (ADG, P > 0.05), gain-to-feed ratio (G:F, P > 0.05), the incidence of diarrhea (P > 0.05), or the antioxidant capacity (P > 0.05) between two groups. The expression of IL-1ß and TNF-α in jejunal mucosa were significantly decreased (P < 0.05) in piglets fed the LC-supplemented diet in comparison to the control. CONCLUSION: The results of this study indicate that dietary supplementation with LC at 50 mg/kg was effective for enhancing the growth performance in weaned piglets, improving intestinal barrier function and alleviating intestinal inflammation.
Subject(s)
Antioxidants/metabolism , Chitosan/pharmacology , Cytokines/metabolism , Intestine, Small/drug effects , Swine/growth & development , Zonula Occludens-1 Protein/metabolism , Animals , Diet/veterinary , Intestine, Small/anatomy & histology , Real-Time Polymerase Chain Reaction/veterinary , Swine/anatomy & histology , WeaningABSTRACT
Interleukin (IL)-22-producing Natural Killer (NK) cells protect the gut epithelial cell barrier from pathogens. A strain of probiotics, Lactobacillus plantarum (L. plantarum, LP), was previously found by our laboratory to significantly improve the mucosal barrier integrity and function of the small intestine in pigs. However, it was unclear whether LP benefited the intestinal mucosal barrier via interactions with the intestinal NK cells. The present study, therefore, was focused on the therapeutic effect of NK cells that were stimulated by LP on attenuating enterotoxigenic Escherichia coli (ETEC)-induced the damage to the integrity of the epithelial cell barrier. The results showed that LP can efficiently increase protein levels of the natural cytotoxicity receptor (NCR) family, and the expression levels of IL-22 mRNA and protein in NK cells. Transfer of NK cells stimulated by LP conferred protection against ETEC K88-induced intestinal epithelial barrier damage in NCM460 cells. We found that NK cells stimulated by LP could partially offset the reduction in NCM460 cell monolayers transepithelial electrical resistance (TEER) caused by ETEC K88, and increase ZO-1 and occludin mRNA and protein expressions by ETEC K88-infected NCM460 cells. Furthermore, adding NK cells that were stimulated by LP to ETEC K88-infected NCM460cells, IL-22R1, p-Stat3, and p-Tyk2 expression by NCM460 cells was increased. Mechanistic experiment showed that NK cells stimulated by LP lost the function of maintaining TEER of NCM460 cells challenged with ETEC K88, when polyclonal anti-IL-22 antibody was used to block IL-22 production. Collectively, our results suggested that LP stimulation of NK could enhance IL-22 production, which might be able to provide defense against ETEC-induced damage to the integrity of intestinal epithelial barrier.
Subject(s)
Interleukins/metabolism , Intestinal Mucosa/immunology , Killer Cells, Natural/immunology , Lactobacillus/immunology , Cell Line , Enteropathogenic Escherichia coli/pathogenicity , Humans , Interleukins/genetics , Intestinal Mucosa/microbiology , Occludin/genetics , Occludin/metabolism , Tight Junctions/metabolism , Interleukin-22ABSTRACT
Pidotimod is a synthetic dipeptide with biological and immunological activity in innate immune responses. It has been reported that pidotimod could promote functional maturation of dendritic cells, but little is known about the regulation of macrophages. Recent studies have demonstrated that M1 or M2 polarized macrophages are of great importance for responses to microorganism infection or host mediators. The aim of this study was to determine the effectiveness of pidotimod on mouse bone marrow-derived macrophage polarization and its function. The results showed that pidotimod had no influence on M1-polarized macrophage. While interestingly, a significant increase of M2 marker gene expression (Arg1, Fizz1, Ym1, MR) was observed (p < 0.01) in IL-4-induced M2 macrophage treated with pidotimod. In addition, cell surface expression of mannose receptor was dramatically enhanced using fluorescence activated cell sorter (FACS) analysis. Furthermore, the function of M2 macrophage was also determinated. The results showed that the supernatant of pidotimod-treated M2 macrophage could increase the migration (p < 0.05) and enhance the wound closure rate (p < 0.05) of MLE-12 cells. Collectively, it could be concluded that pidotimod significantly facilitated IL-4-induced M2 macrophage polarization and improves its function.
Subject(s)
Cell Polarity/drug effects , Macrophages/drug effects , Macrophages/immunology , Peptides/pharmacology , Pyrrolidonecarboxylic Acid/analogs & derivatives , Thiazolidines/pharmacology , Animals , Biomarkers/analysis , Macrophage Activation/drug effects , Macrophages/cytology , Mice , Mice, Inbred C57BL , Pyrrolidonecarboxylic Acid/pharmacologyABSTRACT
BACKGROUND: Long-term physical exercise has been shown to benefit patients with Parkinson disease (PD), but there is a lack of evidence regarding the underlying mechanism. A better understanding of how such benefits are induced by exercise might contribute to the development of therapeutic targets for improving the motor function in individuals with PD. The purpose of this study was therefore to investigate the possible association between exercise-induced motor improvements and the changes in serum microRNA (miRNA) levels of PD patients through small RNA sequencing for the first time. METHODS: Thirteen PD patients completed our 3-month home-and-community-based exercise program, while 6 patients were assigned to the control group. Motor functions were measured, and small RNA sequencing with data analysis was performed on serum miRNAs both before and after the program. The results were further validated by quantitative real-time polymerase chain reaction. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were then conducted to determine the role of differentially expressed miRNAs. RESULTS: The 3-month home-and-community-based exercise program induced significant motor improvements in PD patients in terms of Unified Parkinson's Disease Rating Scale activities of daily living and Motor Subscale (Pâ <â .05), comfortable walking speed (Pâ =â .003), fast walking speed (Pâ =â .028), Six-Minute Walk Test (Pâ =â .004), Berg Balance Scale (Pâ =â .039), and Timed Up and Go (Pâ =â .002). A total of 11 miRNAs (10 upregulated and one downregulated) were identified to be remarkably differentially expressed after intervention in the exercise group, but not in the control group. The results of miRNA sequencing were further validated by quantitative real-time polymerase chain reaction. It was found that the targets of altered miRNAs were mostly enriched in the mitogen-activated protein kinase, Wnt, and Hippo signaling pathways and the GO annotations mainly included binding, catalytic activity, and transcription regulator activity. CONCLUSION: The exercise-induced motor improvements were possibly associated with changes in circulating miRNA levels in PD patients. These miRNAs, as well as the most enriched pathways and GO terms, may play a critical role in the mechanism of exercise-induced benefits in PD and serve as novel treatment targets for the disease, although further investigations are needed.
Subject(s)
MicroRNAs , Parkinson Disease , Humans , MicroRNAs/metabolism , Parkinson Disease/genetics , Parkinson Disease/therapy , Activities of Daily Living , Walking , Physical Therapy ModalitiesABSTRACT
Sensitive detection of pro-gastrin-releasing peptide (Pro-GRP) is crucial because it is a highly sensitive and specific tumor marker for small cell lung cancer. Herein, we synthesized an efficient luminescent europium metal-organic framework and developed a sandwich ECL immunosensor for the sensitive detection of Pro-GRP, which used Eu3+ as the central ion and 2,4,6-tri (4-carboxyphenyl)-1,3,5-triazine (H3TATB) as the organic ligand. H3TATB acted as a strong absorbing reagent and transferred its energy to Eu3+ via the antenna effect to enhance the ECL response signal of Eu3+. As per calculations, the ECL efficiency of Eu-TATB, which was a promising ECL luminophore, was up to 130 %. The Cu2O cube worked as a substrate to assist the electron transfer and was used as a co-reaction accelerator to catalyze S2O82- to produce more SO4â¢- and then enhance the ECL intensity of Eu-TATB. Under optimal experimental conditions, the ECL immunosensor had a linear range of 5 fg mL-1-50 ng mL-1 for detecting Pro-GRP with a detection limit of 1.6 fg mL-1; moreover, it demonstrated excellent stability and specificity and has been successfully applied for detecting Pro-GRP in the human serum.
Subject(s)
Biosensing Techniques , Metal Nanoparticles , Metal-Organic Frameworks , Humans , Gastrin-Releasing Peptide , Europium , Luminescent Measurements , Electrochemical Techniques , Immunoassay , Limit of DetectionABSTRACT
Oxidative stress has been associated with a number of physiological problems in swine, including reduced production efficiency. Recently, although there has been increased research into regulatory mechanisms and antioxidant strategies in relation to oxidative stress-induced pig production, it remains so far largely unsuccessful to develop accurate models and nutritional strategies for specific oxidative stress factors. Here, we discuss the dose and dose intensity of the causes of oxidative stress involving physiological, environmental and dietary factors, recent research models and the antioxidant strategies to provide theoretical guidance for future oxidative stress research in swine.
ABSTRACT
Depending on the microenvironment, macrophages can acquire distinct functional phenotypes, referred to as classically activated M1 and M2. M1 macrophages are considered potent effector cells that kill intracellular pathogens, and M2 macrophages promote the resolution of wound healing. In this study, we are interested to know whether probiotic Bacillus amyloliquefaciens (Ba) can induce macrophages polarization. Real-time fluorescence PCR analysis demonstrated that the expression of IL-1ß, iNOS, TNF-α and IL-6 genes for M1 macrophages was significantly increased at 1.5 h after probiotic Ba treatment compared to the probiotic Ba-free treatment (P < 0.01), whereas the expression of M2 macrophage marker genes (Arg1, Fizz1, MR, Ym1) was decreased (P < 0.05). Furthermore, the phagocytic activity was dramatically increased in the Ba-treated BMDMs using a FITC-dextran endocytosis assay. Together, these findings indicated that probiotic Ba facilitated polarization of M1 macrophages and enhanced its phagocytic capacity. The results expanded our knowledge about probiotic function-involved macrophage polarization.
Subject(s)
Bacillus , Macrophage Activation , Macrophages/immunology , Probiotics/administration & dosage , Transcriptome , Animals , Bacillus/classification , Bacillus/metabolism , Macrophages/cytology , Macrophages/metabolism , Mice , Mice, Inbred C57BL , Nitric Oxide/metabolismABSTRACT
Probiotic is a preparation containing microorganisms that confers beneficial effect to the host. This work assessed whether oral administration of Bacillus amyloliquefaciens SC06 (Ba) could decrease bacterial translocation in weaned mice. Weaned C57BL/6 were randomly allocated into three groups: group I as the control group, group II were treated with 0.85 % NaCl. Group III was administered with probiotic Ba 1 × 10(9) CFU/day dissolved in 100 µl of 0.85 % NaCl for 30 days. Mice were then sacrificed, and tissue were cultured to determine bacterial translocation. Meanwhile, splenic CD4(+)T cells, CD8(+)T cells, B cells, and macrophages were analysised by FACS. Our results showed that probiotic Ba significantly reduced bacteria translocation compared with the control group and 0.85 % NaCl group (P < 0.05), lower levels of bacteria were detected in the MLN, liver, spleen, and kidney of mice. Moreover, significant increase in percentage and number of macrophages were observed in the spleen of Ba-treated mice compared with the control and 0.85 % NaCl groups. Together, these data indicated that Ba could decrease bacterial translocation in weaned mice. This effect seems to be correlated with the changes of macrophage numbers.
ABSTRACT
Piglet weaning is an important stage in production where changes in the environment and diet can cause problems such as intestinal inflammation and diarrhea. Vitamin A is an essential nutrient for human and animal growth and has immunomodulatory and inflammatory effects. A large body of literature has previously reported on the use of vitamin A in piglet production, so our experiment added different concentrations of vitamin A (0, 1100, 2200, 4400, 8800, and 17,600 IU/kg) to weaned piglet diets to study the effects of different doses on growth performance, intestinal barrier, inflammation, and flora in weaned piglets. We selected 4400 IU/kg as the optimum concentration of vitamin A in relation to average daily weight gain, feed intake, feed-to-weight ratio, and diarrhea rate, and subsequently tested the inflammatory factors, immunoglobulin content, antioxidant levels, and intestinal flora of weaned piglets. Results: We observed that the diarrhea rate of weaned piglets was significantly lower after the addition of 4400 IU/kg of vitamin A to the diet (p < 0.05). A control group and a 4400 IU/kg VA group were selected for subsequent experiments. We found that after the addition of vitamin A, the serum CAT level of weaned piglets increased significantly, the expression of Claudin-1 in the jejunum and ileum increased significantly, the expression of Occludin gene in the jejunum increased significantly, the expression of IL-5 and IL-10 in the ileum increased significantly (p < 0.05), and the expression of IL-4, IL-5, and IL-10 in the ileum increased significantly (p < 0.05). Meanwhile, in the colonic flora of vitamin A-added weaned piglets, the relative abundance of Actinobacteria and Erysipelotrichales decreased significantly, while the relative abundance of Bacteroidales increased significantly (p < 0.05). The results of this study indicated that vitamin A at 4400 IU/kg reduces diarrhea in weaned piglets by increasing antioxidant levels, increasing intestinal tight junction protein gene expression, and regulating colonic gut microbiota.
ABSTRACT
This study aimed to introduce an immunosensor for measuring amyloid ß 42 (Aß42) levels by aggregation-induced enhanced electrochemiluminescence (ECL). Metal-organic gels (MOGs) are novel soft materials with advantages such as high gel stability, good light-emitting properties, and easy preparation. This study used silver nanoparticle metal-organic gel (Ag-MOG) as a substrate to connect Aß42-Ab2 and the cathodoluminescent probe. Potassium persulfate was used as a co-reactant that could emit a high ECL signal. CuS@Au had the benefits of a relatively large surface area with excellent carrier function; therefore, it was used as a substrate to load a large amount of Aß42-Ab1, significantly improving the immunosensor sensitivity. The ECL intensity of Aß42 was linear in the range of 0.01 pg/mL to 250 ng/mL with a detection limit of 2.2 fg/mL (S/N = 3) under optimized detection conditions. This ECL immunosensor has been successfully applied to detect Aß42 in human serum with the advantages of excellent stability and high selectivity. This method not only expands the potential applications of ECL immunosensors based on biological testing and clinical diagnosis but also provides a viable approach to basic clinical testing.
Subject(s)
Amyloid beta-Peptides , Electrochemical Techniques , Luminescent Measurements , Metal Nanoparticles , Peptide Fragments , Silver , Amyloid beta-Peptides/analysis , Amyloid beta-Peptides/blood , Silver/chemistry , Humans , Metal Nanoparticles/chemistry , Luminescent Measurements/methods , Peptide Fragments/analysis , Peptide Fragments/blood , Peptide Fragments/chemistry , Biosensing Techniques/methods , Limit of Detection , Immunoassay/methods , Gold/chemistry , Copper/chemistryABSTRACT
Introduction: More effective and environment-friendly organic trace minerals have great potential to replace the inorganic elements in the diets of livestock. This study aimed to investigate the effects of dietary replacement of 100% inorganic trace minerals (ITMs) with 30-60% organic trace minerals (OTMs) on the performance, meat quality, antioxidant capacity, nutrient digestibility, and fecal mineral excretion and to assess whether low-dose OTMs could replace whole ITMs in growing-finishing pigs' diets. Methods: A total of 72 growing-finishing pigs (Duroc × Landrace × Yorkshire) with an initial average body weight of 74.25 ± 0.41 kg were selected and divided into four groups with six replicates per group and three pigs per replicate. The pigs were fed either a corn-soybean meal basal diet containing commercial levels of 100% ITMs or a basal diet with 30, 45, or 60% amino acid-chelated trace minerals instead of 100% ITMs, respectively. The trial ended when the pigs' weight reached ~110 kg. Results: The results showed that replacing 100% ITMs with 30-60% OTMs had no adverse effect on average daily gain, average daily feed intake, feed/gain, carcass traits, or meat quality (P > 0.05) but significantly increased serum transferrin and calcium contents (P < 0.05). Meanwhile, replacing 100% ITMs with OTMs tended to increase serum T-SOD activity (0.05 ≤ P < 0.1), and 30% OTMs significantly increased muscle Mn-SOD activity (P < 0.05). Moreover, replacing 100% ITMs with OTMs tended to increase the apparent digestibility of energy, dry matter, and crude protein (0.05 ≤ P < 0.1) while significantly reducing the contents of copper, zinc, and manganese in feces (P < 0.05). Discussion: In conclusion, dietary supplementation with 30-60% OTMs has the potential to replace 100% ITMs for improving antioxidant capacity and nutrient digestibility and for reducing fecal mineral excretion without compromising the performance of growing-finishing pigs.
ABSTRACT
This experiment was conducted to evaluate effects of zine oxide (ZnO) and condensed tannins (CT), independently or in combination, on the growth performance and intestinal health of weaned piglets in enterotoxigenic Escherichia coli (ETEC-K88)-challenged environment. Randomly divided 72 weaned piglets into 4 groups. Dietary treatments included the following: basic diet group (CON), 1,500 mg/kg zinc oxide group (ZnO), 1,000 mg/kg condensed tannins group (CT), and 1,500 mg/kg zinc oxide +1,000 mg/kg condensed tannins group (ZnO + CT). Dietary ZnO supplementation decreased diarrhea rate from 0 to 14 days, 15 to 28 days, and 0 to 28 days (p < 0.05) and no significant on growth performance. The effect of CT on reducing diarrhea rate and diarrhea index was similar to the results of ZnO. Compared with the CON group, ZnO increased the ileum villus height and improved intestinal barrier function by increasing the content of mucin 2 (MUC-2) in jejunum and ileum mucosa and the mRNA expression of zonula occludens-1 (ZO-1) in jejunum (p < 0.05) and the expression of Occludin in duodenum and ileum (p < 0.05). The effects of CT on intestinal barrier function genes were similar to that of ZnO. Moreover, the mRNA expression of cystic fibrosis transmembrane conductance regulator (CFTR) in jejunum and ileum was reduced in ZnO group (p < 0.05). And CT was also capable of alleviating diarrhea by decreasing CFTR expression and promote water reabsorption by increasing AQP3 expression (p < 0.05). In addition, pigs receiving ZnO diet had higher abundance of phylum Bacteroidetes, and genera Prevotella, and lower phylum Firmicutes and genera Lactobacillus in colonic contents. These results indicated that ZnO and CT can alleviate diarrhea and improve intestinal barrier function of weaned pigs in ETEC-challenged environment. In addition, the application of ZnO combined with CT did not show synergistic effects on piglet intestinal health and overall performance. This study provides a theoretical basis for the application of ZnO in weaning piglet production practices, we also explored effects of CT on the growth performance and intestinal health of weaned piglets in ETEC-challenged environment.
ABSTRACT
Intestinal epithelial barrier injury disrupts immune homeostasis and leads to many intestinal disorders. Lactobacillus reuteri (L. reuteri) strains can influence immune system development and intestinal function. However, the underlying mechanisms of L. reuteri LR1 that regulate inflammatory response and intestinal integrity are still unknown. The present study aimed to determine the effects of LR1 on the ETEC K88-induced intestinal epithelial injury on the inflammatory response, intestinal epithelial barrier function, and the MLCK signal pathway and its underlying mechanism. Here, we showed that the 1 × 109 cfu/ml LR1 treatment for 4 h dramatically decreased interleukin-8 (IL-8) and IL-6 expression. Then, the data indicated that the 1 × 108 cfu/ml ETEC K88 treatment for 4 h dramatically enhanced IL-8, IL-6, and tumor necrosis factor-α (TNF-α) expression. Furthermore, scanning electron microscope (SEM) data indicated that pretreatment with LR1 inhibited the ETEC K88 that adhered on IPEC-J2 and alleviated the scratch injury of IPEC J2 cells. Moreover, LR1 pretreatment significantly reversed the declined transepithelial electrical resistance (TER) and tight junction protein level, and enhanced the induction by ETEC K88 treatment. Additionally, LR1 pretreatment dramatically declined IL-8, IL-17A, IL-6, and TNF-α levels compared with the ETEC K88 group. Then, ETEC K88-treated IPEC-J2 cells had a higher level of myosin light-chain kinase (MLCK), higher MLC levels, and a lower Rho-associated kinase (ROCK) level than the control group, while LR1 pretreatment significantly declined the MLCK and MLC expression and enhanced ROCK level in the ETEC K88-challenged IPEC-J2 cells. Mechanistically, depletion of MLCK significantly declined MLC expression in IPEC-J2 challenged with ETEC K88 compared to the si NC+ETEC K88 group. On the other hand, the TER of the si MLCK+ETEC K88 group was higher and the FD4 flux in the si MLCK+ETEC K88 group was lower compared with the si NC+ETEC K88 group. In addition, depletion of MLCK significantly enhanced Claudin-1 level and declined IL-8 and TNF-α levels in IPEC-J2 pretreated with LR1 followed by challenging with ETEC K88. In conclusion, our work indicated that L. reuteri LR1 can decline inflammatory response and improve intestinal epithelial barrier function through suppressing the MLCK signal pathway in the ETEC K88-challenged IPEC-J2.
Subject(s)
Enterotoxigenic Escherichia coli , Escherichia coli Infections , Intestinal Mucosa , Limosilactobacillus reuteri , Animals , Cell Line , Enterotoxigenic Escherichia coli/immunology , Escherichia coli Infections/metabolism , Interleukin-6/metabolism , Interleukin-8/metabolism , Intestinal Diseases/microbiology , Intestinal Mucosa/microbiology , Intestinal Mucosa/physiology , Limosilactobacillus reuteri/physiology , Myosin-Light-Chain Kinase/metabolism , Signal Transduction , Swine , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Background: Resveratrol has numerous beneficial properties, including antioxidant, anti-inflammatory, and immunomodulatory properties. High summer temperatures in Southern China affect the reproductive performance of sows. The present study aimed to investigate the effects of dietary resveratrol supplementation in different thermal environments on the reproductive performance, antioxidant capacity, immune function, and intestinal microbes of sows and piglets during late gestation and lactation, as well as their relationship with colostrum immunoglobulin. Methods: A two-phase experiment was conducted with 40 healthy multiparous sows. In the first phase of the experiment, 20 sows were used in a moderate temperature environment, and in the second phase of the experiment, the remaining 20 sows were used in a high-temperature environment. In both phases, sows were fed either a control diet or a diet consists of control diet and 300 mg/kg resveratrol starting on day 75 of gestation. Plasma, milk, and fecal samples were collected to obtain the indices of antioxidant capacity, immune function, and intestinal microbes. Results: The results showed that resveratrol supplementation increased the number of live births by 13.24 and 26.79% in the first and second phases, respectively, compared with the control group. In the second phase, resveratrol supplementation increased litter weight at weaning and in the concentrations of growth hormone (GH), insulin (INS), progesterone (PROG), triglycerides, and uric acid (UA). The plasma superoxide dismutase (SOD) level on day 110 of gestation and day 14 of lactation, as well as glutathione peroxidase (GSH-Px) on day 14 of lactation in the first phase, showed an increasing trend (p = 0.0728, p = 0.0932, and p = 0.067, respectively) in the resveratrol group, compared with the control group. On day 14 of lactation, the plasma total antioxidant capability (T-AOC) level was higher in the second phase, while the plasma malondialdehyde (MDA) level was lower in both phases in the resveratrol group. Resveratrol supplementation increased the abundance of immunoglobulin A (IgA), immunoglobulin G (IgG), and immunoglobulin M (IgM) in colostrum and the relative abundance of Lactobacillus and Alloprevotella but decreased the relative abundance of Escherichia-shigella in piglet feces in the second phase. In addition, Spearman's correlation analysis indicated that the weight gain of weaned piglets was positively (p < 0.05) associated with IgM content in colostrum and the abundance of Lactobacillus in the fecal microbiota of piglets in the second phase. Moreover, the abundance of Alloprevotella was positively correlated with the contents of IgA and IgG in colostrum, while the abundance of Lactobacillus was positively correlated with IgM content. Conclusion: These findings indicated that maternal resveratrol supplementation could enhance the growth performance, antioxidant capacity, and intestinal health of piglets in a high temperature environment, which might be associated with increased immunoglobin secretion from colostrum.
ABSTRACT
This study aimed to investigate responses of the Lactobacillus reuteri or an antibiotic on cecal microbiota and intestinal barrier function in different stages of pigs. A total of 144 weaned pigs (Duroc × Landrace × Yorkshire, 21 days of age) were randomly assigned to the control group (CON, fed with a basal diet), the antibiotic group (AO, fed with basal diet plus 100 mg/kg olaquindox and 75 mg/kg aureomycin), and the L. reuteri group (LR, fed with the basal diet + 5 × 1010 CFU/kg L. reuteri LR1) throughout the 164-d experiment. A total of 45 cecal content samples (5 samples per group) from different periods (14th, 42th, and 164th days) were collected for 16S rRNA gene amplification. The results revealed that although LR and AO did not change the diversity of cecal microbiota in pigs, the abundance of some bacteria at the genus level was changed with age. The proportion of Lactobacillus was increased by LR in early life, whereas it was decreased by AO compared with the control group. The relative abundance of Ruminococcaceae was increased along with age. In addition, the gas chromatography results showed that age, not AO or LR, has significant effects on the concentrations of SCFAs in the cecum of pigs (P < 0.05). However, the mRNA expression of tight junction proteins zonula occluden-1 (ZO-1) and occludin were increased by AO in the cecum of pigs on day 14, while LR increased the mRNA expression of intestinal barrier-related proteins ZO-1, occludin, mucin-1, mucin-2, PG1-5, and pBD2 in the cecum of pigs on days 14 and 164 (P < 0.05). In conclusion, LR and AO have different effects on the intestinal barrier function of the cecum, and neither LR nor AO damaged the intestinal barrier function of pig cecum. In addition, LR and AO have little effects on cecal microflora in different stages of the pigs. The microflora and their metabolite SCFAs were significantly changed along with age. These findings provide important information to understand the homeostasis of the cecum of pigs after antibiotic or probiotic treatment.