BW Lipopolysaccharide Research Articles

Threonine attenuates lipopolysaccharide-induced intestinal inflammatory responses in rabbits.

Threonine (Thr) can be involved in the synthesis of immunoglobulins, which play the role of immune regulation, Thr also has to improve intestinal morphology, adjust the sticky protein synthesis, maintain the intestinal barrier function, etc. The experiment aimed to investigate the effects of diets supplemented with different levels of Thr on growth performance and intestinal health of rabbits under lipopolysaccharide (LPS) stress conditions. A total of 180 healthy 35-day-old weaned New Zealand White rabbits were randomly assigned in a 2 × 3 factorial design to receive an intraperitoneal injection of 100µg/kg BW LPS or saline and three diets with different levels of digestible threonine (0.43%, 0.54%, and 0.64%). The LPS challenge resulted in a reduction in body weight in rabbits at day 22, as well as a decrease in the serum d-lactic acid (D-LA) content and the number of goblet cells (GCs) in the jejunum. Additionally, the duodenum JAM2 and JAM3 were down-regulated. The expression of OCLN, ZO-1, and IL-2 in the jejunum, and CLDN, nuclear factor-κB (NF-κB) and ZO-1 mRNA in the ileum were also down-regulated. Furthermore, the duodenum TLR4 and IL-1β mRNA expression, while the jejunum exhibited an elevation in CLDN, TNF-α, and ileum TNF-α mRNA expression (P < 0.05). In the context of LPS challenge condition, dietary Thr addition was found to down-regulate the duodenum ZO-1 and jejunum CLDN mRNA expression of rabbits (P < 0.05). This was accompanied by an increase in ileum sIgA content and GCs number (P < 0.05). Additionally, dietary Thr addition resulted in a downregulation of duodenum TLR4, MyD88, NF-κB, TNF-α and IL-1β, jejunum MyD88, and IL-1β mRNA expression, as well as an up-regulation of ileum IL-10 mRNA expression in rabbits (P < 0.05). In conclusion, the LPS challenge can result in intestinal inflammation and damage the integrity of the intestinal barrier in rabbits. Nevertheless, dietary Thr supplementation can alleviate the intestinal inflammatory response in rabbits challenged with LPS.

Effect of supplementation with Lactobacillus rhamnosus GG powder on intestinal and liver damage in broiler chickens challenged by lipopolysaccharide.

This study explores the effect of dietary along with Lactobacillus rhamnosus GG (LGG) powder on intestinal and liver damage in broiler chickens challenged by lipopolysaccharide (LPS). A total of 100 healthy 1-day-old Ross 308 broiler chickens were selected and randomly divided into two treatments: the control group and the LGG treatment group. There were five replicates for each group, with 10 chickens per replicate. The chickens in the control group were fed a basal diet, while LGG treatment was supplemented with 1,000 mg/kg LGG along with the basal diet. The experiment lasted 29 days, and the trial included two phases. During the first 27 days, the animals were weighed on the 14th and 27th days to calculate growth performance. Then, on day 29, 2 animals from each replicate were intraperitoneally injected with 1 mg/kg BW LPS, and another 2 animals were treated with an equal volume of saline. The chickens were slaughtered 3 h later for sampling and further analysis. (1) LGG addition to the diet did not affect growth performance, including average daily gain (ADG), average daily feed intake (ADFI), and feed-to-weight ratio (F/G) of broiler chickens; (2) LPS stimulation decreased villus height (VH), and caused oxidative stress and increased the amount of diamine oxidase (DAO) in plasma, and the relative expression of intestinal inflammation genes (interleukin-8 [IL-8], interleukin 1β [IL-1β], inducible nitric oxide synthase [iNOS], and tumor necrosis factor-α [TNF-α]) and the relative expression of liver injury genes (b-cell lymphoma 2 [BCL2], heat shock protein70 [HSP70], and matrix metallopeptidase 13 [MMP13]). (3) Supplementation of LGG increased VH and the relative expression of intestinal barrier genes (mucins 2 [Mucin2] and occludin [Occludin]) and decreased the amount of DAO in plasma and the relative expression of intestinal inflammatory factors (IL-8, iNOS, and IL-1β). LGG supplementation also increased the expression of liver injury-related genes (MMP13 and MMP9). In conclusion, LGG enhanced intestinal barrier function, improved intestinal morphology, and alleviated the intestines' inflammatory response in LPS-stimulated broiler chicken, and it has a slightly protective effect on liver damage.

The activation of TLR4-MyD88 signaling promotes hepatic dysfunction and fibrotic changes in SD rats resulting from prolonged exposure to sodium arsenite

Arsenic, a poisonous metalloid element, is linked to liver diseases, but the exactmechanisms for this process are not yet to be completely elucidated. Toll like receptor 4 (TLR4), acting as a pathogenic pattern recognition receptor, plays a pivotal role in various inflammatory diseases via the myeloid differentiation factor 88 (MyD88) pathway. This study aims to investigate the involvement of the TLR4-MyD88 signaling pathway in liver injury induced by prolonged exposure to sodium arsenite (NaAsO2) in Sprague-Dawley rats. Our research findings demonstratethe activation of TLR4-MyD88 signaling pathway in long-term NaAsO2-exposed rat liver tissues, leading to a significant release of inflammatory factors, which suggests its potential involvement in the pathogenesis of NaAsO2-induced liver injury. We further administered lipopolysaccharide (LPS), a natural ligand of TLR4, and TAK-242, a specific inhibitor of TLR4, to rats in order to validate the specific involvement of the TLR4-MyD88 signaling pathway in NaAsO2-induced liver injury. The results showed that, 1 mg/kg.bw LPS treatment significantly activated TLR4-MyD88 signalling pathway and its mediated pro-inflammatory factors, leading to up-regulation of activation indicators in hepatic stellate cells (HSCs) as well as increased secretion levels of extracellular matrix (ECM) in the liver, and ultimately induced liver fibrosis and dysfunction in rats. Relevantly, subsequent administration of 0.5 mg/kg.bw TAK-242 significantly attenuated the expression levels of TLR4 and its associated proteins, mitigated collagen deposition, and partially improved liver fibrosis and dysfunction caused by NaAsO2 in rats. Our study fully confirms the pivotal role of the TLR4-MyD88 signaling in promoting liver injury induced by NaAsO2, thereby providing a novel molecular target for preventing and treating patients with arsenic poisoning-related liver injury.

Disrupted Insulin Signaling Due to an Acute Immune Response in Swine

Insulin is an anabolic hormone involved in glucose uptake and synthesis of fats, proteins, and glycogen. Altered insulin signaling can occur due to metabolic state, pathogen exposure, and autoimmune disorders. Lipopolysaccharide (LPS), an endotoxin secreted produced by gram-negative bacteria, can induce a severe, systemic immune response. In pigs, chronic LPS exposure has induced insulin resistance. However, the effects of acute exposure to LPS on insulin signaling and resistance has not been elucidated. Crossbred nursery pigs (58, 12.1 kg ± 0.23kg BW) were randomly assigned into two experimental subsets, where 10 animals were designated for repeated blood sampling and 48 animals underwent timed euthanasia for tissue collection. For repeated blood sampling, jugular catheters were placed under anesthesia and the animals were given 48 hours to recover before being randomly assigned to treatments. These treatments were an intraperitoneal injection of either 15μg/kg BW LPS or an equal volume of 0.9% Saline (n=5 pigs/treatment). Blood was collected for 48 hours followed by euthanasia. The other 48 pigs were randomly assigned to the same treatments, and then randomly assigned to euthanasia timepoints of 0, 3, 6, 12, 24 or 48 hours post injection for tissue collection. Among the catheterized pigs, two hours post LPS challenge there was a decrease in serum insulin concentration (p&lt;0.05) and mild hyperglycemia (p&lt;0.05) post challenge. At 12 and 24 hours post LPS challenge there was a decrease in insulin (p&lt;0.05) and a trend hyperglycemia was also apparent at 24 hours (p=0.063). In liver tissue, insulin related genes exhibited alterations post LPS injection. Phosphoinositide 3-kinase ( pi3k) and insulin receptor substrate 1 ( irs1) expression were decreased at 3 and 6 hours (p&lt;0.05) and protein kinase b ( akt) expression were decreased at 3 hours (p&lt;0.05) post challenge. The expression of insulin inhibitor growth factor receptor bound protein 10 ( grb10) was decreased at 6, 12, and 24 hours after challenge (p&lt;0.05). Fructose-1-6-bisphosphatase ( f16bp) expression was reduced at 3, 6, and 12 hours (p&lt;0.05) in the liver while phosphoenolpyruvate carboxykinase ( pepck) expression was decreased at 3 and 6 hours (p&lt;0.01) post challenge. Other metabolic genes were decreased at 3, 6, and 12 hours post LPS challenge, including fatty acid synthase ( fas) (p&lt;0.05), carbohydrate response element binding protein ( chrebp) (p&lt;0.05), and acetyl CoA carboxylase ( acc)(p&lt;0.05). Protein abundance of Akt 3 hours after LPS injection was decreased in the liver (p&lt;0.05). In skeletal muscle 6 hours post LPS injection, insulin receptor ( insr), insulin-like growth factor receptor ( igfr) and fas were all increased (p&lt;0.05) These results indicate disrupted insulin signaling at various levels after an acute immune response. Research reported in this abstract was supported by Eunice Kennedy Shriver National Institute of Child Health &amp; Human Development (NICHD) of the National Institutes of Health under award number R01HD092304A. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

The effect of repeated lipopolysaccharide endotoxin challenge on immune response of breeding ewes and subsequent lamb performance.

Infectious disease caused by exposure to Gram-negative bacterial endotoxin lipopolysaccharide (LPS) is recognized to suppress female fertility. However, the effect of varying low-dose endotoxin exposure during distinct stages of follicle development on immune response, reproductive performance, and lamb performance has yet to be elucidated. Therefore, the objective of this study was to evaluate acute phase response, mRNA abundance of inflammatory markers, reproductive performance and lamb growth characteristics of ewes challenged with subclinical doses of LPS. Rambouillet ewes (n = 36; 68.2±1.1kg; age 3 to 7 yr) stratified by body weight (BW) and age were assigned to treatment groups. Ewes received subcutaneous injections of saline (CON, n = 12), 1.5 µg/kg BW LPS (LOW, n = 12), or 3.0 µg/kg BW LPS (HIGH, n = 12) on days 5, 10, and 15 of a synchronized follicular wave. Ewes were subsequently placed with a raddle-painted ram on day 16 for a 35-d breeding season. On treatment days 5 and 15, blood samples, peripheral blood leukocytes, and rectal temperature were collected before and at regular intervals for 12h after LPS challenge. Immune response to LPS was confirmed by increased temperature and serum cortisol concentrations on days 5 and 15. Endotoxin increased circulating plasma concentration of the acute phase protein, haptoglobin by greater than 15%, in both LPS-treated groups on days 5 and 15 at 12h compared with control (P≤ 0.05). Pro- and anti-inflammatory mRNA gene expression demonstrated no differences in expression for tumor necrosis factor-α or peroxisome proliferator-activated receptor gamma among treatment groups (P > 0.10). Likewise, Toll-like receptor 4 (TLR4), interleukin-8 (IL-8), and superoxide dismutase 2 (SOD2) expression was similar among treatment groups on day 5. However, ewes challenged with LPS on day 15 displayed greater mRNA expression for TLR4 from 2 to 6h (P < 0.05), a 7-fold increase for IL-8 from 1.5 to 2.5h (P < 0.05), and 8-fold induction for SOD2 from 2 to 6h (P < 0.05) as compared with controls. First service conception rates were 90% for control ewes and 75% for both treated groups (P = 0.84). Treated ewes demonstrated a reduction in lamb birth weight compared with controls (P ≤ 0.05) and a tendency for reduction of 60-d adjusted weaning weight (P = 0.09). Data suggest that subacute endotoxin exposure aligning with key follicle and oocyte maturation events results in detrimental growth performance of the subsequent lamb.

Ethyl acetate extract of fungus comb from Malayan termite (Macrotermes gilvus Hagen) mound modulates splenic inflammatory responses in mice

Background: The fungus comb is a unique structure inside termites’ nests that facilitates the growth of Termitomyces sp. as a nutrient source for the termites. It is known to possess immunomodulatory properties that boost the immune system. Aim: This objective of this study was to assess the impact of ethyl acetate extract of fungus comb (EAEFC) on the inflammatory reaction in the spleen of mice induced by intraperitoneal injection of lipopolysaccharide (LPS). Methods: An experimental study was conducted using a post-test-only control group design with male BALB/C mice (n=24). The mice were divided randomly into four groups, each comprising six mice, and administered substances via gavage. Groups I and III were administered a solution of 5% dimethyl sulfoxide (DMSO) in distilled water, while Groups II and IV were given 500 mg/kg BW EAEFC dissolved in 5% DMSO. On the fifteenth day, Groups I and II received intraperitoneal injection of 5 ml/kg BW saline, while Groups III and IV were injected with 10 mg/kg BW lipopolysaccharide (LPS) dissolved in saline. After three hours, the mice were euthanized and splenic immunohistology was examined under a light microscope. The results were expressed as mean ± standard deviation (SD), while the group differences were assessed statistically. Results: The expression of interleukin (IL)-1, furin, and activated NK cell was significantly higher in the inflamed model after EAEFC supplementation, while the extract suppressed IL-10. Conclusion: EAEFC was found to alter cytokine expression in the spleen in response to inflammation.

396 Implications of Repeated Subacute Lipopolysaccharide Exposure During Synchronized Estrous

Abstract Acute inflammation induced by chronic infection or a single intravenous administration of the bacterial endotoxin, lipopolysaccharide (LPS) induces immune responses capable of depressing female fertility. However, the effect of dose and repeated endotoxin exposure during the estrous cycle on immune response and reproductive performance has yet to be investigated. To investigate the effects of varying doses of LPS delivered at distinct stages of follicular maturation, thirty-six Ramboulliet ewes (68.2 ± 1.1 kg; age 3 to 7 yr) were stratified by weight and age and randomly assigned to vehicle (n = 12), low (n = 12) or high (n = 12) LPS-treated groups to evaluate febrile response, serum progesterone and cortisol concentrations, and reproductive performance. Ewes received subcutaneous injections of saline (CON), 1.5 µg/kg BW LPS (LOW), or 3.0 µg/kg BW LPS (HIGH) on d 5, 10, and 15 after onset of synchronized estrus. Estrous synchronization was determined by serum progesterone (P4) analysis. All ewes demonstrated similar P4 profiles, but HIGH-LPS ewes tended to have elevated values (P = 0.1). On treatment d 5 and 15, blood samples and rectal temperature were collected before and intermittently throughout the duration of the trial. Lipopolysaccharide increased rectal temperature of LOW and HIGH groups at 2, 3, and 6 h, respectively after initial LPS challenge (d 5) when compared with vehicle-treated controls (0.67, and 0.70 °C, P &amp;lt; 0.05), but no significance was observed at d 10 or 15. Serum concentrations of cortisol were similar among treatment groups at 0 h (P &amp;gt; 0.05). However, a rapid and sustained cortisol increase was demonstrated within 1 h of LPS exposure in both LPS-treatment groups compared with controls (P = 0.001). A similar pattern of cortisol release among treatment groups was demonstrated on d 15 (P &amp;lt; 0.05). While there was no significant difference in services per cycle by treatment, there was a marked difference (16.67%) in remarks during the second cycle for HIGH and LOW groups (P = 0.23) as compared with controls. Control ewes had d 100 pregnancy rate of 90.91%, while low and high LPS-treated groups had pregnancy rates of 91.67% (P = 0.96) and 83.33% (P = 0.59) respectively. The administration of LPS resulted in significant increases in serum cortisol in treated ewes, and serum progesterone was significantly increased in LOW LPS-treated ewes compared with CON or HIGH LPS-treated ewes. Thus, alterations of temperature, steroid hormone concentration, and time to estrus in response to LPS may ultimately implicate female fertility and prevent economic losses for producers.

2-(3-(chloromethyl)benzoyloxy)benzoic Acid Increases CD4+ Regulatory T-Cell Population and FoxP3 Expression in Lipopolysaccharide-induced Mice

BACKGROUND: Lipopolysaccharide (LPS) has been reported to increase CD4+ regulatory T-cell (CD4+ Treg) populations. Acetylsalicylic acid (ASA) has been reported to have immunomodulatory activity, but it may induce chronic gastric ulceration. Another salicylic acid-bearing compound, 2-(3-(chloromethyl)benzoyloxy)benzoic acid (3-CH2Cl), has been reported to have less gastric mucosal damage. However, the effect of 3-CH2Cl on CD4+ Tregs in LPS-induced mice is still unknown. Therefore, the present study was conducted to investigate the immunomodulatory effect of 3-CH2Cl on CD4+ T-cell and CD4+ Treg populations as well as FoxP3 expression in LPS-induced mice.METHODS: Synthesis of 3-CH2Cl was performed by mixing salicylic acid and chloromethylbenzoylchloride with the catalyzation of pyridine, acetone and heat. The 3-CH2Cl tablets were prepared using direct compression method. After intraperitoneal injection of 1 mg/kg BW LPS to mice, 60 mg/kg BW ASA or 60 mg/kg BW 3-CH2Cl was given orally for 3 days. The splenocyte was obtained through splenectomy and collagenase digestion. The population of CD4+ T-cells and CD4+ Tregs, as well as the splenic FoxP3 expression were determined using flow cytometry technique.RESULTS: CD4+ T-cell populations in mice treated with LPS and 3-CH2Cl or ASA were lower than those treated with LPS merely. Meanwhile, CD4+ Treg populations and FoxP3 expression levels in mice treated with LPS and 3-CH2Cl or ASA were higher than those treated with LPS merely.CONCLUSION: Since 3-CH2Cl could decrease CD4+ T-cell population and increase CD4+ Treg population mediated by the increase of FoxP3 expression in LPS-induced inflammation, it may act as a potential therapeutic drug to reduce inflammatory conditions.KEYWORDS: 2-(3-(chloromethyl)benzoyloxy)benzoic acid, acetylsalicylic acid, CD4, T-regulatory cells, FoxP3, LPS

Dietary tryptophan supplementation enhances mitochondrial function and reduces pyroptosis in the spleen and thymus of piglets after lipopolysaccharide challenge.

The thymus and spleen, the main reservoirs for T lymphocytes, modulate the innate immune response. Oxidative stress, excessive inflammation and abnormal pyroptosis can cause dysfunction of these organs. This study aimed to examine whether tryptophan supplementation can improve growth performance and mitochondrial function via the adenosine 5'-monophosphate-activated protein kinase (AMPK)/sirtuin1 (Sirt1)/peroxisome proliferator-activated receptor-γ coactivator-1α (PGC1α) signalling pathway and decrease pyroptosis via the nucleotidebinding oligomerisation domain-like receptor protein 3 (NLRP3)/caspase-1/gasderminD (GSDMD) signalling pathway in the spleen and thymus of piglets after lipopolysaccharide (LPS) challenge. Eighteen weaned piglets were allotted to three treatment groups: non-challenged control, LPS-challenged control and LPS+0.2% tryptophan. On day 35, the pigs in the LPS and LPS+0.2% tryptophan groups were injected with 100μg/kg BW LPS, whereas those in the control group were administered with sterile saline. At 4h postchallenge, the weaned piglets were sacrificed, and their thymuses and spleens were collected. Results showed that tryptophan enhanced growth performance and antioxidant status by increasing catalase, glutathione peroxidase and total superoxide dismutase activities and decreasing malondialdehyde and reactive oxygen species contents. Tryptophan also reduced the mRNA levels of proinflammatory cytokine genes and enhanced mitochondrial function by increasing the mRNA levels of mitochondrial transcription factor A, nuclear respiratory factor-1, mitochondria transcription factor B1, AMPKα1, AMPKα2, Sirt1 and PGC1α and the protein expression of phosphorylated AMPK, Sirt1 and PGC1α. It also reduced pyroptosis by decreasing the mRNA levels of NLRP3, apoptosis-associated speck-like protein containing CARD, caspase-1 and GSDMD and the protein expression of NLRP3, caspase-1 and GSDMD. These results indicate that tryptophan supplementation enhances growth performance and mitochondrial function via the AMPK/Sirt1/PGC1α signalling pathway and decreases pyroptosis via the NLRP3/caspase-1/GSDMD signalling pathway in the spleen and thymus of LPS-challenged piglets.

Epidermal growth factor activates EGFR/AMPK signalling to up-regulate the expression of SGLT1 and GLUT2 to promote intestinal glucose absorption in lipopolysaccharide challenged IPEC-J2 cells and piglets

Epidermal growth factor (EGF) plays an important role in nutrients transport. The present study was to investigate the effects of EGF on glucose absorption in a cellular injury model (IPEC-J2 cells, in vitro study) and animal injury model (weaned piglets, in vivo study) established by lipopolysaccharide (LPS). In vitro study: porcine intestinal epithelial cells (IPEC-J2) were divided into four treatments: control group (Control); 100 ng/mL EGF treatment group (EGF); 1 μg/mL LPS treatment group (LPS) and 100 ng/mL EGF plus 1 μg/mL LPS treatment group (EGF + LPS). The results showed that EGF significantly increased the alkaline phosphatase (AKP) and sodium/potassium-transporting adenosine triphosphatase (Na+/K+-ATPase) activity, and significantly improved the mRNA and protein expression of SGLT1, GLUT2, EGF receptor (EGFR) and AMP-activated protein kinase α1 (AMPK-α1) in LPS-induced injured cells. In vivo experiment: twenty-four piglets weaned at 21 days were randomly assigned into: (1) control group (basal diets); (2) EGF group (basal diet +2 mg/kg EGF); (3) LPS group (basal diet + injection with 100 μg/kg BW LPS) and (4) EGF + LPS group (basal diet + 2 mg/kg EGF + injection with 100 μg/kg BW LPS). Our results showed that EGF significantly increased the AKP and Na+/K+-ATPase activity, and significantly improved the mRNA expression of SGLT1, GLUT2, EGFR and AMPK-α1 in jejunum mucosa of piglets challenged with LPS. In conclusion, EGF can activate EGFR/AMPK signalling to up-regulate the expression of SGLT1 and GLUT2 as well as improve the AKP and Na+/K+-ATPase activity, thereby promoting intestinal glucose absorption in IPEC-J2 cells and piglets challenged by LPS. HIGHLIGHTS EGF promotes SGLT1 and GLUT2 expression and AKP and Na+/K+-ATPase activity in LPS-induced injured porcine intestinal epithelial cells. EGF promotes SGLT1 and GLUT2 expression and AKP and Na+/K+-ATPase activity in jejunum mucosa of piglets challenged by LPS. Dietary supplementation with EGF activates EGFR/AMPK signalling to up-regulate the expression of SGLT1 and GLUT2 as well as improve the AKP and Na+/K+-ATPase activity to promote intestinal glucose absorption in lipopolysaccharide challenged piglets.

Maternal immune activation and dietary soy isoflavone supplementation influence pig immune function but not muscle fiber formation.

The goals of this study were to determine the impact of maternal PRRSV infection on offspring muscle and immune development and the potential of dietary soy isoflavones to mitigate those effects. Thirteen first-parity gilts ("gilts") were randomly allotted into one of three treatments: not infected and fed a diet devoid of isoflavones (CON), infected with porcine reproductive and respiratory syndrome virus (PRRSV) and fed the control diet (POS) or that supplemented with 1,500 mg/kg soy-derived isoflavones (ISF). Gilts were inoculated with PRRSV intranasally on gestational day (GD) 70. After farrowing (GD 114 ± 2), 1-2 offspring ("pigs") closest to the average litter weight were selected either at birth (3 ± 2 d of age) or weaning (21 ± 2 d of age) to determine body, muscle, and organ weights as well as muscle cell number and size. Four weaned pigs of average body weight within each litter were selected for postnatal immune challenge. At PND 52, pigs were injected with 5 µg/kg BW lipopolysaccharide (LPS) intraperitoneally. Serum was collected at 0, 4, and 8 h following LPS administration to analyze tumor necrosis factor alpha (TNF-α). At PND 59, pigs were administered a novel vaccine to elicit an adaptive immune response. At PND 59, 66, and 73, peripheral blood mononuclear cells were isolated and T-cell populations determined by flow cytometry. Both POS and ISF pigs exhibited persistent PRRSV infections throughout the study (PND 1-73). At PND 3, whole body, muscle, and organ weights were not different (P > 0.22) between groups, with the exception of relative liver weight, which was increased (P < 0.05) in POS compared with CON pigs. At PND 21, ISF pigs had reduced (P ≤ 0.05) whole body and muscle weights, but greater (P < 0.05) kidney weight compared with CON, and greater (P < 0.05) relative liver weight compared with CON and POS. Muscle fiber number and size were not different (P > 0.39) between groups at birth or weaning. After LPS administration, TNF-α was greatest in ISF pigs (P < 0.05) at both 0 and 8 h post-challenge. At the peak time-point of 4 h post-challenge, ISF pigs had the greatest concentration of TNF-α and CON pigs had the lowest, with POS pigs being intermediate (P = 0.01). After vaccination, ISF offspring had shifts in T-cell populations indicating an impaired immune response. These data indicate that maternal PRRSV infection may impact offspring organ growth and immune function, particularly when the dam is supplemented with isoflavones.

The effects of deoxynivalenol-contaminated corn in low-complexity diets supplemented with either an immune-modulating feed additive, or fish oil on nursery pig growth performance, immune response, small intestinal morphology, and component digestibility.

Three hundred twenty newly weaned pigs (21 days of age; 6.7 ± 0.3 kg BW) were used to determine the effects of supplementing low-complexity (LC) deoxynivalenol- (DON) contaminated nursery diets with a feed additive or fish oil on growth performance and immune response to an Escherichia coli lipopolysaccharide (LPS) challenge. Pens were randomly assigned to 1 of 5 dietary treatments (n = 8 pens per treatment): positive control (PC; contained multiple animal protein sources), or 1 of 4 LC diets (contained only plant-based protein sources) without (NC; negative control) or with ~ 3.5 ppm DON contamination, without (DON‐) or with a feed additive containing a blend of immune-modulating components (DON+; 2 mg/kg, as-fed) or fish oil (DONω3; 2.5%, as-fed). Dietary treatments were fed during phases I and II (7 and 15 days, respectively) and a common phase III diet was fed for 20 days. On day 22, two pigs per pen were injected IM with 30 μg/kg BW LPS and 1 pig per pen with 1 mL saline. Rectal temperatures were recorded at 0, 1, 2, 3 h after injection. At 3 h, blood was collected for plasma cytokine analysis and small intestinal histomorphology was assessed. In phase I, pigs fed PC and NC did not differ for ADG, ADFI and G:F, but these outcomes were greater than for pigs fed DON+ and DONω (P < 0.05). In phase II, pigs fed NC had greater ADG and PC had greater ADFI but lower G:F than pigs fed DON‐ and DONω3 (P < 0.05). At the end of phase II, pigs fed DONω3 tended to have lower BW than PC and NC (P = 0.084 and 0.079, respectively). In phase III and overall, there were no differences among dietary treatments for ADG, ADFI, G:F, or final BW. The LPS injection increased rectal temperature and reduced jejunal and ileal villus height (versus saline; P < 0.05). Plasma interferon-γ concentration was only increased by LPS for pigs fed PC, NC, and DON+ compared to the saline-injected counterparts (P < 0.05). Regardless of LPS injection, jejunal villus height was greater for pigs fed DON+ than DONω3 (P < 0.05) and ileal villus height was greater for pigs fed DON+ and PC than DONω3 (P < 0.05). Therefore, nursery diet complexity did not affect growth performance or immune response to LPS. Regardless of DON contamination and feed additive inclusion in phases I and II, pigs were able to achieve nursery exit BW not different from those fed PC. The feed additive offered marginal benefits for small intestinal villus height and immune response for pigs fed DON-contaminated LC nursery diets.

Prenatal immune stimulation alters the postnatal acute phase and metabolic responses to an endotoxin challenge in weaned beef heifers.

This study evaluated whether administration of lipopolysaccharide (LPS) at each trimester of gestation would alter the acute phase (APR) and metabolic responses to a postnatal LPS challenge in weaned heifers. Pregnant crossbred multiparous cows (n = 50) were randomized into prenatal immune stimulation (PIS; n = 24; administered 0.1 µg/kg BW LPS subcutaneously at 71 ± 2, 170 ± 2 and 234 ± 2 d of gestation) and saline (CON; n = 26) groups. From these treatment groups, heifer calves (n = 12 PIS and 11 CON) were identified at weaning (244 ± 3 d of age) to receive an LPS challenge. On d 0, heifers were fitted with vaginal temperature (VT) devices, jugular catheters, and moved into individual stalls. On d 1, heifers were challenged i.v. with LPS (0.5 µg/kg BW) at 0 h. Blood samples were collected and sickness behavior scores (SBS) recorded at 0.5 h intervals from -2 to 8 h and at 24 h relative to LPS challenge. Serum was analyzed for cortisol, cytokines, glucose, non-esterified fatty acids (NEFA), and serum urea nitrogen (SUN) concentrations. Baseline VT was lesser in PIS heifers from -11 to -5 h pre-LPS (treatment × time: P < 0.01) compared to the CON; however, the post-LPS VT response did not differ between treatments (P = 0.89). There was a treatment × time interaction (P < 0.01) for SBS with PIS heifers having lesser SBS from 0.5 to 2 h post-LPS compared to CON. There was a treatment × time interaction (P = 0.03) for cortisol with PIS heifers having greater cortisol at 0.5, 3, 3.5, 5.5 and 6.5 h post-LPS compared to CON. There were treatment × time interactions for the post-LPS cytokine responses (P ≤ 0.05). Specifically, PIS heifers had greater TNF-α from 1.5 to 2 h, yet less TNF-α at 3 h than CON (P < 0.01), and PIS heifers had greater IFN-γ from 3.5 to 5.5 h post-LPS than CON (P < 0.01). In contrast, IL-6 was less in PIS than CON heifers from 1.5 to 8 h post-LPS (P < 0.001). Glucose concentrations were greater in PIS heifers at -1 h, but less at 2, 3 and 5.5 h compared to CON (treatment × time: P < 0.01). Serum NEFA concentrations were greater (P = 0.04) in PIS than CON heifers. There was a treatment × time interaction (P < 0.01) for SUN with PIS heifers having greater SUN concentrations at -2, -1.5, 2, 3, 6.5 and 24 h than CON. These data demonstrate that in utero exposure to multiple low doses of endotoxin has lasting physiological and immunological effects when the offspring encounter a similar postnatal immunological insult.

PERBEDAAN SUHU ABDOMEN DAN ANUS PADA MENCIT DENGAN BERBAGAI KONDISI

Research about sepsis in animal model is interesting to get the right therapeutic method for human being. We did the research to study about the value of body temperature in mice’s various body area in order to get the important information if we are going to do the translational research in animal model about sepsis. One clinical signs of the sepsis mice model is changes in body temperature. An easy way to examine body temperature is using infrared non-contact thermometer. This study aimed to compare the body temperature using infrared non-contact thermometer at the abdomen and anal area. We used male mice, weighing 25–30 g, divided into two groups (control and treatment groups). The control group injected with NaCl 0.9% solution, with the amount of NaCl 0.9% volume equal to Lipopolysaccharide (LPS). In the treatment group injected with 2.5 mg/kg BW of LPS intraperitoneally. Body temperature was measured in abdomen (tabd) and ananl (tan) area at 8th and 24th hour after treatment. Body temperature value tabd was higher than tan. Lipopolysaccharide injection increase body temperature but was not significant when compared to the control group (8th and 24th hour). The mean difference between tabd and tan in 8th control groups were 2.12oC respectively. The mean difference between tabd and tan in 24th hour control groups 4.6oC. The mean difference in treatment groups (8th hour) was 4.66oC, while it was 4.77oC in the 24th groups. Giving 2.5 mg/kg BW LPS intraperitoneally did not rise the body temperature significantly as compare to control groups. But, body temperature at anus area using non-contact infrared thermometer after treatment showed lower results as compared to that of at abdomen significantly.

233 NaturSafe® supplementation mitigates some aspects of the acute phase immune response to a lipopolysaccharide (LPS) challenge in weaned beef calves

Abstract This study was conducted to determine if feeding calves NaturSafe would reduce the acute phase response (APR) to lipopolysaccharide (LPS) challenge. Crossbred steers (n=32; 274±2 kg) were randomly allotted to two treatment diets: 1) Control, fed a standard receiving ration, and 2) NaturSafe, fed the Control ration supplemented with NaturSafe at 12 g/hd/d (NaturSafe®, Diamond V). On d22, steers were fitted with indwelling jugular catheters and rectal temperature monitoring devices and placed in individual stalls. On d23, steers were challenged i.v. with 0.25 µg/kg BW LPS. Serum samples were collected and sickness behavior scores (SBS) recorded at 0.5-h intervals from -2 to 8h and at 24h relative to LPS challenge. Rectal temperatures were greater (P=0.01) in NaturSafe compared to Control steers for the following time intervals following LPS challenge: 6 to 11h, 13h, 15 to 20h, and 22 to 24h. Additionally, SBS were reduced (P&amp;lt; 0.01) in NaturSafe compared to Control steers. White blood cell concentrations were greater (P=0.05) in NaturSafe compared to Control steers prior to the LPS challenge, yet the response to LPS did not differ between treatments (P &amp;gt;0.05). A treatment × time interaction for serum cortisol concentrations (P&amp;lt; 0.01) showed an increase at 0.5 and 2h post-challenge but a reduction at 3h in NaturSafe compared to Control steers. Additionally, fibrinogen was greater (P&amp;lt; 0.01) in NaturSafe compared to Control steers. There was a treatment × time interaction (P&amp;lt; 0.01) for TNF-α where concentrations were reduced from 1 to 2h post-challenge in NaturSafe compared to Control steers. Serum IL-6 tended (P=0.09) to show a reduction in serum concentrations in NaturSafe compared to Control steers. There was a tendency (P=0.07) for a treatment × time interaction for IFN-γ. Overall these data suggest a priming effect of NaturSafe on the innate immune system of steers, resulting in an attenuated APR to the LPS challenge.

2 Acute phase response to lipopolysaccharide challenge in beef steers supplemented with prebiotic blends

Abstract A study was conducted to determine the effects of two prebiotic blends on the acute phase response (APR) following lipopolysaccharide (LPS) challenge in steers. Crossbred steers (n = 36; 273±4 kg) were fed for 21d on three different treatments: 1) Control, fed a standard receiving ration; 2) Control ration supplemented with a Prebiotic/Probiotic blend (28.4 g/hd/d; PMI); and 3) Control ration supplemented with a DFM/Prebiotic blend (19.0 g/hd/d; PMI). On d20, calves were fitted with indwelling rectal temperature (RT) monitors and jugular catheters and moved into individual stanchions in a covered barn. On d21, blood samples were collected, and sickness behavior scores recorded at 0.5-h intervals from -2 to 8h and again at 24h relative to an i.v. challenge with 0.25 µg/kg BW LPS. Serum was isolated and stored until analyzed for pro-inflammatory cytokines, cortisol and glucose concentrations. Complete blood counts were measured every 2h from -2 to 8h and again at 24h. Pre-challenge RT measured for 18h prior to the challenge tended (P = 0.10) to be affected by treatment such that calves fed the Prebiotic/Probiotic blend had greater RT than Control and tended to be greater than calves fed the DFM/Prebiotic blend (38.9, 39.2, and 39.0±0.1oC, respectively). Post-challenge RT increased 0.8–1.0oC on average but did not differ between treatments (P = 0.53). Sickness behavior scores were not different between treatments (P = 0.14). There were no differences in white blood cell or differential counts between treatments (P ≥ 0.25). Serum concentrations of TNF-α, IL-6, and IFN-γ increased in response to the challenge (P &amp;lt; 0.01) but were not different between treatments (P ≥ 0.26). Serum cortisol and glucose concentrations were reduced in both supplemented groups compared to Control steers (P ≤ 0.006). Therefore, the data suggest that the effects of the prebiotic blends during an immune challenge were limited to alterations in metabolic biomarkers and energy utilization.

MORG1+/\u2212 mice are protected from histological renal damage and inflammation in a murine model of endotoxemia

BackgroundThe MAPK-organizer 1 (MORG1) play a scaffold function in the MAPK and/or the PHD3 signalling paths. Recently, we reported that MORG1+/− mice are protected from renal injury induced by systemic hypoxia and acute renal ischemia-reperfusion injury via increased hypoxia-inducible factors (HIFs). Here, we explore whether MORG1 heterozygosity could attenuate renal injury in a murine model of lipopolysaccharide (LPS) induced endotoxemia.MethodsEndotoxemia was induced in mice by an intraperitoneal (i.p) application of 5 mg/kg BW LPS. The renal damage was estimated by periodic acid Schiff’s staining; renal injury was evaluated by detection of urinary and plasma levels of neutrophil gelatinase-associated lipocalin and albumin/creatinine ratio via ELISAs. Renal mRNA expression was assessed by real-time PCR, whereas the protein expression was determined by immunohistochemistry or Western blotting.ResultsLPS administration increased tubular injury, microalbuminuria, IL-6 plasma levels and renal TNF-α expression in MORG1+/+ mice. This was accompanied with enhanced infiltration of the inflammatory T-cells in renal tissue and activation of the NF-κB transcription factors. In contrast, endotoxemic MORG1+/− showed significantly less tubular injury, reduced plasma IL-6 levels, significantly decreased renal TNF-α expression and T-cells infiltration. In support, the renal levels of activated caspase-3 were lower in endotoxemic MORG1+/− mice compared with endotoxemic MORG1+/+ mice. Interestingly, LPS application induced a significantly higher accumulation of renal HIF-2α in the kidneys of MORG1+/− mice than in wild-type mice, accompanied with a diminished phosphorylation of IκB-α and IKK α,β and decreased iNOS mRNA in the renal tissues of the LPS-challenged MORG1+/− mice, indicating an inhibition of the NF-κB transcriptional activation.ConclusionsMORG1 heterozygosity protects against histological renal damage and shows anti-inflammatory effects in a murine endotoxemia model through modulation of HIF-2α stabilisation and/or simultaneous inhibition of the NF-κB signalling. Here, we show for the first time that MORG1 scaffold could represent the missing link between innate immunity and inflammation.

Exposure to lipopolysaccharide in utero alters the postnatal metabolic response in heifers.

This study was designed to determine the effect of prenatal lipopolysaccharide (LPS) exposure on the postnatal metabolic response to an LPS challenge in beef heifers. Pregnant, crossbred cows ( = 50) were assigned to a prenatal immune stimulation (PIS; = 25; administered 0.1 µg/kg BW LPS subcutaneously 233 ± 15d of gestation) or saline treatment group (Control; = 25). Birth and weaning BW of calves were collected. There was not (> 0.05) a treatment × gender interaction for birth weight or 205-d adjusted weaning BW. Treatment did not affect (> 0.05) birth BW, but steers and heifers of PIS cows had greater ( < 0.02) 205-d adjusted weaning BW than offspring from Control cows. From the 2 prenatal treatment groups, heifer calves ( = 12 PIS, 11 Control) were identified at weaning (238 ± 15 d of age) to subsequently receive an LPS challenge. On d 0, heifers were fitted with indwelling jugular catheters and were moved into individual pens. On d 1, heifers (fed at 0600 h) were challenged i.v. with LPS (0.5 µg/kg BW) at 0 h (1000 h). Blood samples were collected at 30-min intervals from -2 to 8 h and again at 24 h relative to the LPS challenge. There was a treatment × time interaction ( < 0.01) for cortisol; PIS heifers had greater cortisol from 4 to 6.5 h post-LPS challenge ( < 0.001). There was a treatment × time interaction ( = 0.04) for serum glucose such that glucose was greater ( = 0.01) in PIS than Control heifers at 0.5 h, but was greater in Control than PIS heifers at 2, 4.5, and 7 h post-LPS challenge. This resulted in overall time ( < 0.01) and treatment ( < 0.01) effects such that Control heifers had greater glucose concentrations than PIS heifers. There was a tendency ( = 0.10) for a treatment × time interaction for serum NEFA, such that NEFA was greater in Control than PIS heifers at -2, -1.5, and 7 h relative to the LPS challenge ( ≤ 0.02). Also, there were time ( < 0.01) and treatment effects ( < 0.01) for NEFA with Control heifers having greater NEFA than PIS heifers. Serum BUN was affected by a treatment × time interaction ( < 0.01). Concentrations of BUN were greater in PIS heifers from -1.5 to -1 h, 1 to 2 h, at 4 h, and from 5 to 24 h relative to the LPS challenge. These results demonstrate postnatal growth and the metabolic responses of weaned beef calves can be significantly altered with a single exposure to LPS in utero.

The immunomodulatory effect of Zingiber cassumunar ethanolic extract on phagocytic activity, nitrit oxide and reaxtive oxygen intermediate secretions of macrophage in mice

Immunomodulators could protect the body from a variety of infectious agents and boost immunity. Zingiber cassumunar rhizome or bangle potentially showed as an immunomodulator through increasing of macrophage activity in vitro. The objective of the study was to determine the effect of Z. cassumunar rhizome ethanolic extract on phagocytic activity, nitrite oxide (NO) and reactive oxygen intermediate (ROI) secretions in macrophages in vivo. A total of 200 g of Z. cassumunar rhizome was powdered, macerated in 96% ethanol and evaporated to get concentrated extract. Mice were divided into 5 groups as follow: the normal group was given by water only, the negative control group was given by a 0.94% CMC-Na suspension, the treatment groups were given by 250, 500 and 1000 mg/kgBW, respectively, of Z. cassumunar ethanolic extract. The extract was administered orally for 7 days. On the 8th day the mice were injected intraperitoneally 0.7 mg/kg BW of lipopolysaccharide. Four hours later macrophage was isolated. Furthermore, the determination of the phagocytic activity, NO and ROI secretions levels of macrophage were performed. The treatments of 250, 500 and 1000 mg/kg BW of Z. cassumunar ethanolic extract significantly increase the ROI and NO secretions levels (p<0.05), but did not increase the phagocytic activity (p>0.05) of macrophage. Z. cassumunar ethanolic extract have immunomodulatory effect in vivo.

Effects of immunomodulatory nutrients on growth performance and immune-related gene expression in layer chicks challenged with lipopolysaccharide

Immunomodulatory nutrients alter the immune response to pathogens. This study was conducted to determine the effects of immunomodulatory nutrients on the immune response to a lipopolysaccharide (LPS) challenge of layer chicks fed supplemental corn oil (control; 3%), fish oil (3%), conjugated linoleic acid (CLA; 1%), lutein (0.05 g/kg), or vitamin E (90 I.U./ kg). Four-week-old layer chicks were allotted to 10 treatment groups arranged as a 2 × 5 factorial with 2 sexes and 5 dietary treatments (TRT; 5 replicate pens of 2 male and 2 female chicks per TRT). After a 2-week diet adaptation, all birds were injected intraperitoneal with 1.5 mg/kg BW LPS. Twelve hours post challenge, samples were collected. Fish oil fed birds had greater (P = 0.03) spleen weight (% final BW) than the CLA fed birds. In the liver, the fish oil TRT had higher (P = 0.040) IL-12 expression than the corn oil TRT, but the corn oil TRT had greater (P = 0.001) IL-4 expression than the CLA, lutein, and vitamin E TRT. There was a main effect of sex of the birds on growth parameters at 12 h post LPS challenge in which male birds had greater beginning BW (P < 0.001), final BW (P < 0.001), and greater 12-hour BW loss (P = 0.020) than the female birds, but not relative weight loss. There were also main effects of sex on immune-related gene expression with the females having greater gene expression than the males in the duodenal mucosal scrapings [IL-1β, IL-12, and TLR-4 gene expression (P = 0.026, 0.011, and 0.002, respectively)]; liver [IL-10, IL-4, and iNOS gene expression (P = 0.017, 0.032, and 0.006, respectively)]; and spleen [IL-1β, IL-10, IL-4, and iNOS gene expression (P = 0.001, 0.001, 0.001, and 0.005, respectively)]. Therefore, each immunomodulatory nutrient added to the diets of layer chickens resulted in different immune responses to an LPS challenge.