The ameliorative role of Aloe vera-loaded chitosan nanoparticles on Staphylococcus aureus induced acute lung injury: Targeting TLR/NF-κB signaling pathways.

Background: Acute lung injury (ALI) is a severe condition distinguished by inflammation and impaired gas exchange in the lungs. Staphylococcus aureus, a common bacterium, can cause ALI through its virulence factors. Aloe vera is a medicinal plant that has been traditionally used to treat a variety of illnesses due to its anti-inflammatory properties. Chitosan nanoparticles are biocompatible and totally biodegradable materials that have shown potential in drug delivery systems. Aim: To explore the antibacterial activity of Aloe vera-loaded chitosan nanoparticles (AV-CS-NPs) against S. aureus in vitro and in vivo with advanced techniques. Methods: The antibacterial efficacy of AV-CS-NPs was evaluated through a broth microdilution assay. In addition, the impact of AV-CS-NPs on S. aureus-induced ALI in rats was examined by analyzing the expression of genes linked to inflammation, oxidative stress, and apoptosis. Furthermore, rat lung tissue was scanned histologically. The rats were divided into three groups: control, ALI, and treatment with AV-CS-NPs. Results: The AV-CS-NPs that were prepared exhibited clustered semispherical and spherical forms, having an average particle size of approximately 60 nm. These nanoparticles displayed a diverse structure with an uneven distribution of particle sizes. The maximum entrapment efficiency of 95.5% ± 1.25% was achieved. The obtained findings revealed that The minimum inhibitory concentration and minimum bactericidal concentration values were determined to be 5 and 10 ug/ml, respectively, indicating the potent bactericidal effect of the NPs. Also, S. aureus infected rats explored upregulation in the mRNA expression of TLR2 and TLR4 compared to healthy control groups. AV-CS-NP treatment reverses the case where there was repression in mRNA expression of TLR2 and TLR4 compared to S. aureus-treated rats. Conclusion: These NPs can serve as potential candidates for the development of alternative antimicrobial agents.

Clinacanthus nutans leaf extract reduces pancreatic ß-cell apoptosis by inhibiting JNK activation and modulating oxidative stress and inflammation in streptozotocin-induced diabetic rats.

Background: Controlling apoptosis induced by oxidative stress in pancreatic ß-cells provides promising strategies for preventing and treating diabetes. Clinacanthus nutans leaves possess bioactive constituents with potential antioxidant and anti-diabetic properties. Aim: This study aimed to investigate the molecular mechanisms by which C. nutans extract protects pancreatic ß-cells from apoptotic damage in streptozotocin (STZ)-induced diabetic rats. Methods: Diabetes was induced in male Wistar rats by intraperitoneal injection of 45 mg/kg STZ, followed by 28 days of treatment with C. nutans leaf extract and Glibenclamide as the standard drug. At the end of the study, blood samples were collected to measure glucose levels, oxidative stress markers, and inflammation. Pancreatic tissue was stained immunohistochemically to detect c-Jun N-terminal kinase (JNK) and Caspase-3 expression. Results: The administration of C. nutans leaf extract to diabetic rats significantly reduced fasting blood glucose, malondialdehyde, and tumor necrosis factor-α levels, while concurrently enhancing the activity of superoxide dismutase. The immunohistochemical studies revealed a decrease in the expression of JNK and caspase-3 in the pancreatic islets of diabetic rats. Conclusion: Clinacanthus nutans exhibits the potential to protect pancreatic ß-cells from apoptosis by suppressing oxidative stress and inflammation.

From growth promotion to intestinal inflammation alleviation: Unraveling the potential role of Lactobacillus rhamnosus GCC-3 in juvenile grass carp (Ctenopharyngodon idella).

Lactobacillus rhamnosus is a probiotic, which not only promotes the growth of animals, but also has anti-inflammatory effects. However, the mechanism by which Lactobacillus rhamnosus regulates intestinal immunity is not well comprehended. Hence, the study aimed to research how Lactobacillus rhamnosus affects the intestinal immunity using juvenile grass carp (Ctenopharyngodon idella) as a model. We selected 1800 juvenile grass carp for testing. They were divided into six treatments and fed with six gradients of Lactobacillus rhamnosus GCC-3 (0.0, 0.5, 1.0, 1.5, 2.0, 2.5 g/kg) for 70 days. Enteritis was subsequently induced with dextroside sodium sulfate. Results indicated that dietary Lactobacillus rhamnosus GCC-3 addition improved growth performance. Meanwhile, appropriate levels of Lactobacillus rhamnosus GCC-3 alleviated excessive inflammatory response by down-regulating the expression of TLR4 and NOD receptors, up-regulating the expression of TOR, and then down-regulating the expression of NF-κB. Additionally, appropriate Lactobacillus rhamnosus GCC-3 improved intestinal immunity by reducing pyroptosis triggered by NLRP3 inflammasome and mediated by GSDME. Furthermore, 16 S rRNA sequencing showing appropriate levels of Lactobacillus rhamnosus GCC-3 increased Lactobacillus and Bifidobacterium abundance and decreased Aeromonas abundance. These results suggest that Lactobacillus rhamnosus GCC-3 can alleviate intestinal inflammation through down-regulating NF-κB and up-regulating TOR signaling pathways, as well as by inhibiting pyroptosis.

Polysaccharide of Atractylodes macrocephala Koidz alleviates LPS-induced proliferation, differentiation inhibition and excessive apoptosis in chicken embryonic myogenic cells.

BACKGROUND: Lipopolysaccharide (LPS) can induce systemic inflammation and affect the growth and development of poultry. As a kind of traditional Chinese medicine, polysaccharide of Atractylodes macrocephala Koidz (PAMK) can effectively improve the growth performance of animals and improve the immunity of animal bodies. OBJECTIVES: The purpose of this study was to investigate the effects of PAMK on LPS-induced inflammatory response, proliferation, differentiation and apoptosis of chicken embryonic myogenic cells. METHODS: We used chicken embryonic myogenic cells as a model by detecting EdU/MYHC immunofluorescence, the expression of inflammation, proliferation, differentiation-related genes and proteins and the number of apoptotic cells in the condition of adding LPS, PAMK, belnacasan (an inhibitor of Caspase1) or their combinations. RESULTS: The results showed that LPS stimulation increased the expression of inflammatory factors, inhibited proliferation and differentiation, and excessive apoptosis in chicken embryonic myogenic cells, and PAMK alleviated these adverse effects induced by LPS. After the addition of belnacasan (inhibitor of Caspase1), apoptosis in myogenic cells was inhibited, and therefore, the number of apoptotic cells and the expression of pro-apoptotic genes Caspase1 and Caspase3 were increased. In addition, belnacasan inhibited the increased expression of inflammatory factors, inhibited proliferation, differentiation and excessive apoptosis in chicken embryonic myogenic cells induced by LPS. CONCLUSIONS: This study provides a theoretical basis for further exploring the mechanism of action of PAMK and exogenous LPS on chicken embryonic myogenic cells and lays the foundation for the development and application of green feed additives in animal husbandry industry.

The azole biocide climbazole induces oxidative stress, inflammation, and apoptosis in fish gut.

Climbazole is an azole biocide that has been widely used in formulations of personal care products. Climbazole can cause developmental toxicity and endocrine disruption as well as gut disturbance in aquatic organisms. However, the mechanisms behind gut toxicity induced by climbazole still remain largely unclear in fish. Here, we evaluate the gut effects by exposing grass carp (Ctenopharyngodon idella) to climbazole at levels ranging from 0.2 to 20 µg/L for 42 days by evaluating gene transcription and expression, biochemical analyses, correlation network analysis, and molecular docking. Results showed that climbazole exposure increased cyp1a mRNA expression and ROS level in the three treatment groups. Climbazole also inhibited Nrf2 and Keap1 transcripts as well as proteins, and suppressed the transcript levels of their subordinate antioxidant molecules (cat, sod, and ho-1), increasing oxidative stress. Additionally, climbazole enhanced NF-κB and iκBα transcripts and proteins, and the transcripts of NF-κB downstream pro-inflammatory factors (tnfα, and il-1ß/6/8), leading to inflammation. Climbazole increased pro-apoptosis-related genes (fadd, bad1, and caspase3), and decreased anti-apoptosis-associated genes (bcl2, and bcl-xl), suggesting a direct reaction to apoptosis. The molecular docking data showed that climbazole could form stable hydrogen bonds with CYP1A. Mechanistically, our findings suggested that climbazole can induce inflammation and oxidative stress through CYP450s/ROS/Nrf2/NF-κB pathways, resulting in cell apoptosis in the gut of grass carp.

Zinc glycinate alleviates LPS-induced inflammation and intestinal barrier disruption in chicken embryos by regulating zinc homeostasis and TLR4/NF-κB pathway.

The effect of an immune challenge induced by a lipopolysaccharide (LPS) exposure on systemic zinc homeostasis and the modulation of zinc glycinate (Zn-Gly) was investigated using a chicken embryo model. 160 Arbor Acres broiler fertilized eggs were randomly divided into 4 groups: CON (control group, injected with saline), LPS (LPS group, injected with 32 µg of LPS saline solution), Zn-Gly (zinc glycinate group, injected with 80 µg of zinc glycinate saline solution) and Zn-Gly+LPS (zinc glycinate and LPS group, injected with the same content of zinc glycinate and LPS saline solution). Each treatment consisted of eight replicates of five eggs each. An in ovo feeding procedure was performed at 17.5 embryonic day and samples were collected after 12 hours. The results showed that Zn-Gly attenuated the effects of LPS challenge-induced upregulation of pro-inflammatory factor interleukin 1ß (IL-1ß) level (P =0.003). The LPS challenge mediated zinc transporter proteins and metallothionein (MT) to regulate systemic zinc homeostasis, with increased expression of the jejunum zinc export gene zinc transporter protein 1 (ZnT-1) and elevated expression of the import genes divalent metal transporter 1 (DMT1), Zrt- and Irt-like protein 3 (Zip3), Zip8 and Zip14 (P < 0.05). A similar trend could be observed for the zinc transporter genes in the liver, which for ZnT-1 mitigated by Zn-Gly supplementation (P =0.01). Liver MT gene expression was downregulated in response to the LPS challenge (P =0.004). These alterations caused by LPS resulted in decreased serum and liver zinc levels and increased small intestinal, muscle and tibial zinc levels. Zn-Gly reversed the elevated expression of the liver zinc finger protein A20 induced by the LPS challenge (P =0.025), while Zn-Gly reduced the gene expression of the pro-inflammatory factors IL-1ß and IL-6, decreased toll-like receptor 4 (TLR4) and nuclear factor kappa-B p65 (NF-κB p65) (P < 0.05). Zn-Gly also alleviated the LPS-induced downregulation of the intestinal barrier gene Claudin-1. Thus, LPS exposure prompted the mobilization of zinc transporter proteins and MT to perform the remodeling of systemic zinc homeostasis, Zn-Gly participated in the regulation of zinc homeostasis and inhibited the production of pro-inflammatory factors through the TLR4/NF-κB pathway, attenuating the inflammatory response and intestinal barrier damage caused by an immune challenge.

Effects of ß-mannanase supplementation on productive performance, inflammation, energy metabolism, and cecum microbiota composition of laying hens fed with reduced-energy diets.

The objective of this study is to investigate the beneficial effects and underlying mechanism of dietary ß-mannanase supplementation on the productive performance of laying hens fed with metabolic energy (ME)-reduced diets. A total of 448 Hy-Line gray laying hens were randomly assigned to seven groups. Each group had 8 replicates with 8 hens. The groups included a control diet (CON) with a ME of 2750 kcal/Kg, diets reduced by 100 kcal/Kg or 200 kcal/Kg ME (ME_100 or ME_200), and diets with 0.15 g/Kg or 0.2 g/Kg ß-mannanase (ME_100+ß-M_0.15, ME_100+ß-M_0.2, ME_200+ß-M_0.15, and ME_200+ß-M_0.2). The productive performance, egg quality, intestinal morphology, inflammatory response, mRNA expression related to the Nuclear factor kappa B (NF-κB) and AMPK pathway, and cecum microbiome were evaluated in this study. ME-reduced diets negatively impacted the productive performance of laying hens. However, supplementation with ß-mannanase improved FCR, decreased ADFI, and restored average egg weight to the level of the CON group. ME-reduced diets increased the levels of interleukin-1ß (IL-1ß) and IL-6 while decreasing the levels of IL-4 and IL-10 in the jejunum of laying hens. However, dietary ß-mannanase supplementation improved jejunum morphology, reduced pro-inflammatory cytokine concentrations, and increased levels of anti-inflammatory factors in laying hens fed with ME-reduced diets. The mRNA levels of IL-6, IFN-γ, TLR4, MyD88, and NF-κB in the jejunum of ME-reduced diets were significantly higher than that in CON, dietary ß-mannanase supplementation decreased these genes expression in laying hens fed with ME-reduced diets. Moreover, dietary ß-mannanase supplementation also decreased the mRNA levels of AMPKα and AMPKγ, and increased the abundance of mTOR in the jejunum of laying hens fed with ME-reduced diets. Cecum microbiota analysis revealed that dietary ß-mannanase increased the abundance of various beneficial bacteria (e.g., g_Pseudoflavonifractor, g_Butyricicoccus, and f_Lactobacillaceae) in laying hens fed with ME-reduced diets. In conclusion, dietary ß-mannanase supplementation could improve the productive performance of laying hens fed with a ME-reduced diet by improving intestinal morphology, alleviating intestinal inflammation, changing energy metabolism-related signaling pathways, and increasing cecum-beneficial microbiota.

Changes in gene expression in the intestinal mucus of broilers with woody breast myopathy.

Previous work has shown that dietary treatments affect woody breast (WB) incidence differently, which indicates that gut conditions such as gut barrier function, inflammation, and oxidative stress are likely related to WB. In this study, dietary supplementation with antibiotics (bacitracin) or probiotics (Bacillus subtilis) was investigated for their effects on the expression of transcripts related to gut barrier function, inflammation, and oxidative stress in the mucus lining of the jejunum from broilers with or without WB. A split-plot experimental design was used in this study. The dietary treatments served as the main plot factor and the breast muscle condition was the subplot factor. On d 41, jejunum mucus was collected from 1 bird from each of 3 replicate pens in each 3 dietary treatment groups that exhibited WB and an additional bird that contained a normal breast (3 biological replicates/treatment/phenotype; 3 × 3 × 2, total N = 18). Total RNA was extracted using a commercial RNA extraction kit. The expression levels of CLDN1, MUC6, TLR2A, TLR2B, TLR4, IFN-γ, IL-1ß, IL-8L1, IL-10, NOS2, and SOD were determined using 2-step RT-qPCR analysis. The gene expression difference in ΔCt values was determined after normalizing with the chicken 18S rRNA gene. When the significant differences occurred between treatments, the relative fold change was calculated using the ΔΔCt method and the significance level was calculated. The PROC GLM procedure of SAS 9.4 was used, and the level of significance was set at P ≤ 0.05. There were no significant interactive effects between diet and the breast muscle condition on the expression of any of the genes tested. However, birds with WB exhibited higher MUC6 (P < 0.0001) gene expression levels than birds with normal breast muscles. In addition, the expression of SOD decreased in birds that were fed the antibiotic diet when compared to birds that were fed the probiotic diet (P = 0.014). In conclusion, WB identified in broilers tested in the current study is attributed to increased expression of mucin, indicating a correlation between WB incidence and gel-forming mucin secretion and pathogen signaling.

Tea polyphenols alleviate TBBPA-induced inflammation, ferroptosis and apoptosis via TLR4/NF-κB pathway in carp gills.

The extensive application of Tetrabromobisphenol A (TBBPA) leads to the pollution of part of the water environment and brings great safety risks to aquatic animals. As a natural extract, tea polyphenols (TPs) have antioxidant and anti-inflammatory effects. Gills are one of the immune organs of fish and constitute the first line of defense of the immune system. However, it was unclear whether TPs could mitigate TBBPA-induced gills injury. Therefore, an animal model was established to investigate the effect of TPs on TBBPA-induced gills. The results indicated that TBBPA changed the coefficient and tissue morphology of carp gills. In addition, TBBPA induced oxidative stress and inflammation, leading to ferroptosis and apoptosis in carp gills. Dietary addition of TPs significantly improved the antioxidant capacity of carp, effectively inhibited the overexpression of TLR4/NF-κB and its mediated inflammatory response. Moreover, TPs restored iron metabolism, reduced the expression of pro-apoptotic factors thereby alleviating ferroptosis and apoptosis in carp gills. This study enriched the protective effect of TPs and provided a new way to improve the innate immunity of carp.

Dietary effects of microalga Tetraselmis suecica on growth, antioxidant-immune activity, inflammation cytokines, and resistance of Nile tilapia fingerlings to Aeromonas sobria infection.

The dietary effects of the green microalga Tetraselmis suecica (TS) on the growth, digestive enzymes, immune and antioxidant responses, genes expression, and disease resistance of Nile tilapia (Oreochromis niloticus) fingerlings were investigated. This microalga was mixed with the diet' ingredients at doses of 0.0 (the control), 5, 10, 15, and 20 g/kg diet and then fed to fish daily for 84 days. After the feeding trial, fish were experimentally challenged with Aeromonas sobria, infection and fish mortalities were recorded for another 10 days. Dietary TS significantly (p < 0.05) enhanced growth, digestive enzymes activities, and blood proteins, particularly at the level of 15 g/kg diet. Feeding the fish on 15 TS/kg feed exhibited highest mRNA expressions of GH and IGF-1 genes as well as SOD, CAT, and GPx genes compared to other TS groups. Moreover, highest levels of hepatic antioxidant and immune indices were found in the treatment of 15 g TS/kg feed. Significant downregulation of IL-1ß and IL-8 genes expression and significant upregulation of IL-10 gene expression were observed in TS-fed fish, principally in fish groups fed on 15-20 g TS/kg feed. Conversely, hepatic malondialdehyde levels, blood glucose, and the activities of transaminases (ALT and AST) were significantly (p < 0.05) decreased in fish fed with 15-20 g TS/kg diet. Serum bactericidal activity against A. sobria was significantly higher in TS-fed fish groups, and its highest levels were found in treatments of 15-20 g/kg diet. Of interest, the survival rates of fish groups fed diets with 10-20 g TS/kg feed were higher after the challenge with A. sobria infection than the control group. Accordingly, we can conclude that supplementing fish diets with a 15 g TS/kg diet enhanced the growth, antioxidant and immune activities, and resistance of Nile tilapia fingerlings to possible A. sobria infection.

Unlocking the potential of N-acetylcysteine: Improving hepatopancreas inflammation, antioxidant capacity and health in common carp (Cyprinus carpio) via the MAPK/NF-κB/Nrf2 signalling pathway.

N-acetylcysteine (NAC) positively contributes to enhancing animal health, regulating inflammation and reducing stress by participating in the synthesis of cysteine, glutathione, and taurine in the body. The present study aims to investigate the effects of dietary different levels of NAC on the morphology, function and physiological state of hepatopancreas in juvenile common carp (Cyprinus carpio). 450 common carps were randomly divided into 5 groups: N1 (basal diet), N2 (1.5 g/kg NAC diet), N3 (3.0 g/kg NAC diet), N4 (4.5 g/kg NAC diet) and N5 (6.0 g/kg NAC diet), and fed for 8 weeks. The results indicated that dietary 3.0-6.0 g/kg NAC reduced hepatopancreas lipid vacuoles and nuclear translocation, and inhibited apoptosis in common carp. Simultaneously, the activities of hepatopancreas alanine aminotransferase and aspartate aminotransferase progressively increased with rising dietary NAC levels. Dietary NAC enhanced the non-specific immune function of common carp, and exerted anti-inflammatory effects by inhibiting the MAPK/NF-κB signaling pathway. Additionally, dietary 3.0-6.0 g/kg NAC significantly improved the antioxidant capacity of common carp, which was associated with enhanced glutathione metabolism, clearance of ROS and the activation of Nrf2 signaling pathway. In summary, NAC has the potential to alleviate inflammation, mitigate oxidative stress and inhibit apoptosis via the MAPK/NF-κB/Nrf2 signaling pathway, thereby improving hepatopancreas function and health of common carp. The current findings provide a theoretical basis for promoting the application of NAC in aquaculture and ecological cultivation of aquatic animals.

Effects of dietary rumen-protected choline supplementation to periparturient dairy cattle on inflammation and metabolism in mammary and liver tissue during an intramammary lipopolysaccharide challenge.

The objective of this experiment was to examine the effects of supplementation and dose of rumen-protected choline (RPC) on markers of inflammation and metabolism in liver and mammary tissue during an intramammary lipopolysaccharide (LPS) challenge. Parous Holstein cows were blocked by calving month and randomly assigned within block to receive 45 g/d of RPC (20.4 g/d of choline ions; CHOL45), 30 g/d of RPC (13.6 g/d of choline ions; CHOL30), or no RPC (CON) as a top-dress starting 24 d before expected calving until 21 d postpartum. Cows were alternately assigned within treatment group to either receive an intramammary LPS challenge (200 µg in each rear quarter; Escherichia coli O111:B4) or not at 17 DIM (CHOL45, n = 9; CHOL45-LPS, n = 9; CHOL30, n = 11; CHOL30-LPS, n = 10; CON, n = 10; CON-LPS, n = 9). Hepatic and mammary tissues were collected from all cows on d 17 postpartum. Hepatic and mammary tissues were collected at ∼7.5 and 8 h, respectively, after the LPS challenge. An additional mammary biopsy was conducted on LPS-challenged cows (CHOL45-LPS, CHOL30-LPS, and CON-LPS) at 48 h postchallenge. Hepatic and mammary RNA copy numbers were quantified for genes involved in apoptosis, methylation, inflammation, oxidative stress, and mitochondrial function using NanoString technology. Targeted metabolomics was conducted only on mammary tissue samples (both 8 and 48 h biopsies) to quantify 143 metabolites including choline metabolites, amino acids, biogenic amines and derivatives, organic acids, carnitines, and glucose. Hepatic IFNG was greater in CHOL45 as compared with CON in unchallenged cows, suggesting an improvement in type 1 immune responses. Hepatic CASP3 was greater in CHOL45-LPS as compared with CON-LPS, suggesting greater apoptosis. Mammary IL6 was reduced in CHOL30-LPS cows as compared with CHOL45-LPS and CON-LPS (8 and 48 h). Mammary GPX4 and COX5A were reduced in CHOL30-LPS as compared with CON-LPS (8 h), and SDHA was reduced in CHOL30-LPS as compared with CON-LPS (8 and 48 h). Both CHOL30-LPS and CHOL45-LPS cows had lesser mammary ATP5J than CON-LPS, suggesting that dietary RPC supplementation altered mitochondrial function following LPS challenge. Treatment did not affect mammary concentrations of any metabolite in unchallenged cows, and only 4 metabolites were affected by dietary RPC supplementation in LPS-challenged cows. Mammary concentrations of isobutyric acid and 2 acyl-carnitines (C4:1 and C10:2) were reduced in CHOL45-LPS as compared with CHOL30-LPS and CON-LPS. Taken together, reductions in medium- and short-chain carnitines along with an increase in long-chain carnitines in mammary tissue from CHOL45-LPS cows suggests less fatty acid entry into the ß oxidation pathway. Although the intramammary LPS challenge profoundly affected markers for inflammation and metabolism in liver and mammary tissue, dietary RPC supplementation had minimal effects on inflammatory markers and the mammary metabolome.

Retrospective Study of Intertarsal Joint Inflammation in Avian Species From a Zoological Institution.

A retrospective cross-sectional study was performed to measure the prevalence of avian intertarsal inflammation over a 5-year period, identify risk factors, and discuss treatment options. The authors hypothesized that: 1) long-legged birds would be more affected, 2) participation in a bird show would be a significant risk factor, and 3) young animals would be more frequently affected. Thirty-five clinical cases from 9 avian orders were included in the study. Statistical analysis indicated that the orders Ciconiiformes (9/150; 6%, 95% confidence interval [CI]: 3.2-11), Gruiformes (4/132; 3%, 95% CI: 1.2-7.5), and Pelecaniformes (7/152; 2.8%, 95% CI: 1.4-5.6) were significantly more affected than other orders (P < 0.01). Similarly, long-legged birds (21/35) had 9.8 times greater chance (P < 0.001, 95% CI: 4.7-21) of developing the condition compared with other birds. Participation in a free-flight show (22/35) was a significant risk factor (P < 0.001; odds ratio: 7.0, 95% CI: 3.3-15). Mean age at onset of clinical signs was 5.7 years, and being < 2 years-of-age during the study period was not a significant predictor of disease (P = 0.054). The tibial cartilage, a fragile fibrocartilaginous structure, was frequently affected (34%, 12/35). Treatment protocols included anti-inflammatory drugs, analgesic drugs, or both (94%, 33/35), low-level laser therapy (54%, 19/35), joint immobilization (34%, 12/35), intra-articular corticoid injections (20%, 7/35), surgical stabilization (17%, 6/35), physiotherapy (9%, 3/35), intra-articular hyaluronic acid (6%, 2/35) or platelet-rich plasma (3%, 1/35) injections, and chiropractic care (3%, 1/35). Overall recovery rate was 49% (17/35), and the condition was associated with a poor prognosis in chronic cases.

The mechanism of cadmium exposure-induced lymph node necroptosis and inflammation in piglets: Activation of CYP450 through VDR / CREB1 pathway.

Cadmium (Cd) is toxic non-essential heavy metal that precipitates adverse health effects in humans and animals, but the effect of Cd on lymph node toxicity of piglets is still unclear. In order to explore the possible molecular mechanism of Cd toxicity to lymph nodes of piglets, ten 6-week-old male weaned piglets were randomly divided into two groups, C group and Cd group. Group C was fed with basal diet, while group Cd was fed with basal diet supplemented with CdCl2 (20 mg/kg) for 40 days, the pigs were euthanized and the mesenteric, inguinal and submandibular lymph nodes (MLN, ILN, SLN) were collected. The results indicated that Cd could induce the inflammatory cell infiltration, microvascular hemorrhage, microthrombosis and cell necrosis in MLN, ILN and SLN of piglets, induced Cytochrome P450 proteins (CYP1A1、CYP2E1、CYP2A1 and CYP3A2) mRNA levels and the protein levels of Vitamin D receptor (VDR) and cAMP response element binding protein 1 (CREB1). In addition, Cd exposure upregulated the mRNA and protein levels of dynamin-related protein 1 (DRP1), receptor-interacting protein kinase 3 (RIP3), mixed lineage kinase domain-like protein (MLKL), and increased tumor necrosis factor-α (TNFα), interferon-γ (IFNγ), interleukin-2 (IL-2), interleukin-4 (IL-4), cyclooxygenase 2 (COX-2) protein levels, and the damage degree of three kinds of lymph nodes was similar after Cd exposure. In general, these results manifest that Cd exposure regulates VDR/CREB1 pathway, activates CYP450s, induces necroptosis of lymph nodes, and leads to inflammation.

Influence of different heat stress models on nutrient digestibility and markers of stress, inflammation, lipid, and protein metabolism in broilers.

This experiment determined the effects of different HS models and pair-feeding (PF) on nutrient digestibility and markers of stress, inflammation, and metabolism in broilers. Birds (720 total) were allocated into 12 environmentally controlled chambers and reared under thermoneutral conditions until 20 d. Until 41 d birds were exposed to 4 treatments, including: thermoneutral at 24°C (TN-al), daily cyclic HS (12 h at 24 and 12 h at 35°C; cyHS), constant HS at 35°C (coHS), and PF birds maintained at 24°C and fed to equalize FI with coHS birds (TN-coPF). At d 41, ileal digesta were collected to determine nutrient apparent ileal digestibility (AID). Blood, liver, and breast tissues were collected from 8 birds per treatment to determine the mRNA expression of stress, inflammation, and metabolism markers. An additional 8 TN-al birds were sampled after acute HS exposure at 35°C for 4 h (aHS), and 8 cyHS birds were sampled either right before or 4 h after HS initiation. Data were analyzed by 1-way ANOVA and means were separated using Tukey's HSD test. Compared with TN-al birds, AID of nitrogen and ether extract were reduced in coHS birds, and both cyHS and coHS reduced (P < 0.05) AID of total essential amino acids. TNFα and SOD2 expression were increased (P < 0.05) under aHS, coHS, and TN-coPF conditions. IL6 and HSP70 were increased (P < 0.05) under coHS and aHS, respectively. Expression of lipogenic enzymes ACCα and FASN were reduced by coHS and TN-coPF, while coHS increased the lipolytic enzyme ATGL (P < 0.05). IGF1 was lowered in coHS birds, and p70S6K and MyoG were reduced under coHS and TN-coPF (P < 0.05). Interestingly, MuRF1 and MAFbx were increased (P < 0.05) under coHS only. Overall, these results indicate that coHS has a greater impact on nutrient digestibility and metabolism than aHS and cyHS. Interestingly, increased protein degradation during HS appears to be mostly driven by HS per se and not the reduced FI.

Protective effects of dietary dimethyl itaconate supplementation on oxidative stress, inflammation, and apoptosis in broilers under chronic heat stress.

This study was conducted to evaluate the effects of dietary dimethyl itaconate (DI) supplementation on oxidative stress, inflammation, and apoptosis in broilers under chronic heat stress (HS). Twenty-one-day-old male Ross 308 broilers (n = 120) were randomly allocated to 5 groups: a control group, HS group, HS + 50 mg/kg DI group, HS + 150 mg/kg DI group, and HS + 200 mg/kg DI group. The birds in the control group received the basal diets and were maintained at 21 ± 1 °C for 24 h daily. The birds in the HS group and HS + DI groups were raised at 32 ± 1 °C for 8 h daily and received basal diets containing DI at the indicated dose (0, 50, 150, or 200 mg/kg). The results showed that the contents of alanine aminotransferase, aspartate aminotransferase, and malondialdehyde (MDA) in serum were markedly elevated by exposure to chronic HS (P < 0.01), and this elevation was alleviated by 150 and 200 mg/kg DI supplementation (P < 0.05). Chronic HS-induced declines (P < 0.05) in total antioxidant capacity (T-AOC) and activities of peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT) in serum were markedly attenuated after 200 mg/kg DI treatment in broilers (P < 0.05). Moreover, broilers subjected to chronic HS exhibited higher contents of MDA, protein carbonyl, and hydrogen peroxide (P < 0.01), but lower T-AOC and activities of antioxidant enzymes (P < 0.05), as well as reduced inhibition of superoxide and hydroxyl free radicals (P < 0.01) in the liver compared to the control group; these changes were effectively mitigated by treatment with 200 mg/kg DI in broilers (P < 0.05). In addition, 50-200 mg/kg DI effectively ameliorated chronic HS-stimulated upregulation of the mRNA levels of pro-inflammatory mediators in the livers of broilers (P < 0.01). Dietary supplementation with 150 and 200 mg/kg DI significantly alleviated chronic HS challenge-induced upregulation of the mRNA levels of Bcl-2-associated X, caspase 3, and caspase 9 (P < 0.01), but downregulation of Bcl-2 mRNA levels (P < 0.01) in broilers (P < 0.05). Importantly, chronic HS-induced downregulation of the mRNA or protein levels of nuclear factor (erythroid-derived 2)-like 2 (NRF-2), NADPH quinone acceptor oxidoreductase 1 (NQO1), heme oxygenase-1 (HO-1), SOD2, or glutathione-S-transferases (GST) (P < 0.01) was markedly improved by 150 and 200 mg/kg DI (P < 0.05). The above results indicated that DI can ameliorate oxidative stress, inflammation, and apoptosis in broilers under chronic HS.

Global warming has become increasingly severe in recent years, threatening all life forms on Earth. Poultry are particularly susceptible to heat stress (HS) due to their unique physiological features, such as the absence of sweat glands and a high metabolic rate, and HS thus leads to liver injury and high mortality in broilers. Numerous studies have shown that dimethyl itaconate (DI) exerts beneficial effects in the regulation of inflammation, oxidative stress, and nutrient metabolism. However, it remains unclear whether DI can be used as a dietary supplement to prevent oxidative stress, inflammation, and apoptosis in broilers exposed to chronic HS. Here, we found that DI markedly relieved chronic HS-induced liver injury and enhancement of active molecule contents in the livers of broilers. Simultaneously, DI significantly ameliorated chronic HS by enhancing the antioxidative capacity and reducing the expression of pro-inflammatory cytokines and pro-apoptotic factors in broiler liver, which may be achieved through activation of the nuclear factor (erythroid-derived 2)-like 2 signaling pathway. These results may provide sufficient data to support DI as a dietary supplement for controlling diseases associated with chronic HS in broilers.

Probiotics alleviate constipation and inflammation in late gestating and lactating sows.

Constipation and systemic inflammation are common in late pregnant and lactating sows, which cause health problems like uteritis, mastitis, dystocia, or even stillbirth, further influencing piglets' survival and growth. Probiotic supplementation can improve such issues, but the beneficial mechanism of relieving constipation and enhancing gut motility remains underexplored. This study aimed to investigate the effects and mechanism of probiotic supplementation in drinking water to late pregnant sows on constipation, inflammation, and piglets' growth performance. Seventy-four sows were randomly allocated to probiotic (n = 36) and control (n = 38) groups. Probiotic treatment significantly relieved sow constipation, enhanced serum IL-4 and IL-10 levels while reducing serum IL-1ß, IL-12p40, and TNF-α levels, and increased piglet daily gain and weaning weight. Furthermore, probiotic administration reshaped the sow gut bacteriome and phageome structure/diversity, accompanied by increases in some potentially beneficial bacteria. At 113 days of gestation, the probiotic group was enriched in several gut microbial bioactive metabolites, multiple carbohydrate-active enzymes that degrade pectin and starch, fecal butyrate and acetate, and some serum metabolites involved in vitamin and amino acid metabolism. Our integrated correlation network analysis revealed that the alleviation of constipation and inflammation was associated with changes in the sow gut bacteriome, phageome, bioactive metabolic potential, and metabolism.

Association between transfer of passive immunity, health, and performance of female dairy calves from birth to weaning.

The objective of this observational study was to compare calf health, average daily weight gain, and calf mortality considering the proposed categories of transfer of passive immunity (TPI) by the consensus report of Lombard et al. (2020). The consensus report defines 4 categories of passive immunity (excellent, good, fair, and poor) of calves obtained after colostrum ingestion. The association between the 4 TPI categories was analyzed on calf health (i.e., hazards for morbidity and mortality), and average daily weight gain (ADG) of female Holstein Friesian calves during the first 90 d of age. A further aim of this study was to examine the effects of calving-related factors, such as dystocia or winter season, on TPI status. We hypothesized that calves with excellent TPI have greater ADG, lower risks for infectious diseases such as neonatal diarrhea, pneumonia, and omphalitis, and lower mortality rates. This observational study was conducted from December 2017 to March 2021. Blood was collected from 3,434 female Holstein Friesian dairy calves from 1 commercial dairy farm. All female calves aged 2 to 7 d were assessed for TPI status by determination of total solids (TS) in serum via Brix refractometry by the farm personnel once a week. Passive immunity was categorized according to Lombard et al. (2020) with excellent (≥9.4% Brix), good (8.9-9.3% Brix), fair (8.1-8.8% Brix), or poor TPI (<8.1% Brix). For the analysis of ADG and calving ease 492 or 35 calves had to be excluded due to missing data. The distribution of calves according to TPI categories was as follows: 4.8% poor (n = 166), 29.5% fair (n = 1,012), 28.3% good (n = 971), and 37.4% excellent (n = 1,285). From the calving-related factors, parity of the dam, calving ease, birth month, calving assistance by different farm personnel, and day of life for TPI assessment were significantly associated with TS concentration. Out of 3,434 calves, 216 (6.3%) had diarrhea, and 31 (0.9%) and 957 (27.9%) suffered from omphalitis and pneumonia during the first 90 d of life, respectively. Overall, the morbidity during the preweaning period was 32.6% (n = 1,118), and the mortality was 3.1% (n = 107). The ADG was 0.90 ± 0.15 kg with a range of 0.32 to 1.52 kg. The Cox regression model showed that calves suffering from poor TPI tended toward a greater hazard risk (HR) for diarrhea (HR = 1.57, 95% CI: 0.92-2.69) compared with calves with excellent TPI. Calves suffering from TPI had a greater HR for pneumonia (HR = 2.00, CI: 1.53-2-61), overall morbidity (HR = 1.99, CI: 1.56-2.55), and mortality (HR = 2.47, CI: 1.25-4.86) in contrast to excellent TPI. Furthermore, calves with good and fair TPI had significantly greater HR for pneumonia (good TPI: HR = 1.35, CI: 1.15-1.59; fair TPI: HR = 1.41, CI: 1.20-1.65) and overall morbidity (good TPI: HR = 1.26, CI: 1.09-1.47; fair TPI: HR = 1.32, CI: 1.14-1.53) compared with the excellent TPI category. Average daily weight gain during the first 60 d of life was associated with TPI categories. Calves with excellent and good TPI status had ADG of 0.90 ± 0.01 kg/d and 0.92 ± 0.01 kg/d (mean ± SE), respectively. The ADG of calves with fair TPI status was 0.89 ± 0.01 kg/d, and calves suffering from poor TPI had 0.86 ± 0.01 kg/d. Average daily weight gain differed in calves with poor TPI compared with the other categories. Fair and excellent TPI differed additionally from good TPI. We found no statistical difference between the TPI categories fair and excellent. In conclusion, poor TPI was associated with higher morbidity and mortality during the first 90 d of life. Furthermore, calves with fair, good or excellent TPI had greater ADG.

Acer truncatum leaves extract modulates gut microbiota, improves antioxidant capacity, and alleviates lipopolysaccharide-induced inflammation in broilers.

This study investigated the appropriate way of dietary Acer truncatum leaves (ATL) addition, the effect of disease prevention and its mechanism of action. In experiment 1, 192 Arbor Acres broilers were assigned to 4 treatment groups, fed with basal diets containing 2% bran, replacing it with primary and fermented ATL, and additional 0.3% ATL extract to the basal diet for 42 d, respectively. In experiment 2, 144 broilers were assigned to 3 treatment groups for 21-d trial: (1) C-N group, basal diets, and injected with 0.9% (w/v) sterile saline; (2) C-L group, basal diets, and injected with lipopolysaccharide (LPS); (3) T-L group, ATL diets and injected with LPS. In experiment 1, ATL significantly decreased the index of abdominal fat at 42 d (P < 0.05). ATL extract had a better ability to improve antioxidant capacity and reduce inflammatory levels among all treatment groups, which significantly decreased the content of MDA in the liver and ileum mucosa at 21 d, and increased the expression of IL-10 and Occludin in jejunal mucosa at 42 d (P < 0.05). In experiment 2, ATL significantly increased the level of T-AOC in the liver, decreased the expression of NF-κB in the jejunal mucosa and ileum mucosa (P < 0.05), and restored LPS-induced the changed level of CAT in jejunal mucosa, the expression of IL-6, Claudin-1, and ZO-1 in jejunal mucosa and IL-1ß in ileum mucosa (P < 0.05). Analysis of gut microbiota indicated that ATL enhanced the abundances of Bacteroidota and reduced the proportion of Firmicutes (P < 0.05), and the changed levels of T-AOC in body, IL-1ß, IL-6, IL-10, and NF-κB in jejunum mucosa and propionic acid in cecal were associated with gut microbiota. Collectively, our data showed that the extract of ATL had a better antioxidant and anti-inflammatory effects than primality and fermented. Extraction of ATL modulated intestinal microbiota, and had a protective effect on oxidative stress, inflammation, and intestinal barrier function in broilers challenged with LPS.

Dietary resveratrol supplementation on growth performance, immune function and intestinal barrier function in broilers challenged with lipopolysaccharide.

This study discusses the effects of resveratrol (RES) on the productive performance, immune function and intestinal barrier function of broiler chickens challenged with lipopolysaccharide (LPS). Two hundred and forty 1-day-old male Arbor Acres broilers were randomly divided into 4 groups of 6 replicates each, with 10 broilers per replicate. This experiment used a 2 × 2 factorial design with dietary factors (basal diets or basal diets supplemented with 400 mg/kg RES were administered from d 1 to 21) and stress factors (intraperitoneal injection of 0.5 mg/kg BW of saline or LPS at 16, 18 and 20 d of age). The results showed that LPS challenge had a significant adverse effect on average daily gain (ADG) in broilers at 16 to 21 d of age (P < 0.05), whereas the addition of RES to the diet inhibited the LPS-induced decrease in ADG (P < 0.05). RES also alleviated LPS-induced immune function damage in broilers, which was manifested by the decrease of spleen index (P < 0.05) and the recovery of serum immunoglobulin M and ileal secretory immunoglobulin A content (P < 0.05). The LPS challenge also disrupts intestinal barrier function and inflammation, and RES mitigates these adverse effects in different ways. RES attenuated LPS-induced reduction of villus height in the jejunum and ileum of broilers (P < 0.05). LPS also caused an abnormal increase in plasma D-lactic acid levels in broilers (P < 0.05), which was effectively mitigated by RES (P < 0.05). LPS challenge resulted in a significant decrease in mRNA expression of occludin in the intestinal mucosa (P < 0.05), which was mitigated by the addition of RES (P < 0.05). RES significantly decreased the mRNA expression of toll-like receptor 4, nuclear factor kappa-B and tumor necrosis factor alpha in the ileum tissue stimulated by LPS (P < 0.05). Taken together, this study shows that RES exerts its beneficial effect on broilers challenged with LPS by alleviating immune function damage, relieving intestinal inflammation and barrier damage, and thus improving growth performance.