Serum Biochemistry of Turkeys Challenged with Histomonas meleagridis.

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 have not been elucidated. Crossbred nursery pigs [n = 58, initial body weight (BW) = 12.1 kg ± 0.23 kg] 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 h 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 h 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 h post injection for tissue collection. Among the catheterized pigs, 2-h post LPS challenge there was a decrease in serum insulin concentration (P < 0.05) and mild hyperglycemia (P < 0.05) post challenge. At 12- and 24-h post LPS challenge there was a decrease in insulin (P < 0.05) and a trend hyperglycemia was also apparent at 24 h (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 h (p< 0.05) and protein kinase b (akt) expression were decreased at 3 h (P < 0.05) post challenge. The expression of insulin inhibitor growth factor receptor bound protein 10 (grb10) was decreased at 6-, 12-, and 24-h after challenge (P < 0.05). Fructose-1-6-bisphosphatase (f16bp) expression was reduced at 3-, 6-, and 12-h (P < 0.05) in the liver while phosphoenolpyruvate carboxykinase (pepck) expression was decreased at 3- and 6-h (P < 0.01) post challenge. Other metabolic genes were decreased at 3-, 6-, and 12-h post LPS challenge, including fatty acid synthase (fas; P < 0.05), carbohydrate response element binding protein (chrebp; P < 0.05), and acetyl CoA carboxylase (acc; P < 0.05). Protein abundance of Akt 3 h after LPS injection was decreased in the liver (P < 0.05). In skeletal muscle 6 h post LPS injection, insulin receptor (insr), insulin-like growth factor receptor (igfr) and fas were all increased (P < 0.05) These results indicate disrupted insulin signaling at various levels after an acute immune response.

The benefits of selenium (Se) supplementation during pregnancy and lactation in sows on piglet immunocompetence are not yet fully understood. This study aimed to assess piglet immune status when sows were fed Se-deficient or supplemented diets with sodium selenite (SS) or hydroxy-selenomethionine (OH-SeMet). Twelve pregnant sows were randomly assigned to 3 dietary treatments (4 sows each): no supplemental Se (NC), SS at 0.3 mg Se/kg (SS-0.3), OH-SeMet at 0.3 mg Se/kg (SO-0.3). The trial was carried out from gestation day (gd) 84 to weaning on postpartum day (ppd) 21, at which time all the piglets received the same diet having as Se source SS. On gd 98 and 105, sows were vaccinated with hen egg white lysozyme (HEWL). On ppd 0 and 21, 8 piglets/treatment were euthanized to obtain blood and muscle samples for total Se analyses. Serum samples were collected from 8 female piglets/treatment on ppd 0 and 2 to assess antigen-specific antibody titer for HEWL. On ppd 23, 6 weaned female piglets/treatment were intramuscular challenged with E. coli lipopolysaccharide (LPS) and serum IL-10 levels were measured at 24 h pre-challenge and at 2 and 48 h post challenge. At day 21, piglets from sows fed SO-0.3 showed the highest muscle Se deposition (P < 0.001). At ppd 0, no difference for HEWL-specific IgG was found while at ppd 2, SO-0.3 showed the highest HEWL-specific IgG concentration (P < 0.05). At 24 h pre-challenge and 2 h post LPS challenge, SO-0.3 showed a greater IL-10 level as compare to NC and SS-0.3, respectively (P < 0.05); no difference was found for the same parameter at 48 h post challenge. Piglets from SO supplemented sows exhibited stronger early immune response to LPS challenge. This study indicates the importance of sow Se supplementation with a pure organic form of Se such as OH-SeMet to enhance piglets Se and immune status.

This work evaluated the immune response of finisher pigs with divergent feed efficiency to identify immune pathways that were associated with efficiency. Blood gene expression data from a previous trial (Liu et al., 2019) were re-analyzed with novel methods that quantified changes in biological pathways and associated them with animal performance. Data were quality control checked and aligned to the Sus scrofa reference genome (Sscrofa11.1) using Kallisto. The dataset had 32 samples from eight pigs with high efficiency (HE) or low efficiency (LE) that had been injected with lipopolysaccharide (LPS). Blood samples were taken before challenge, and at 2, 6, and 24 h after challenge. After normalization and evaluation of differential gene expression, activation or inhibition of pathways was quantified using a topology-based pathway activation method and a customization of the Reactome database. Pathways were associated with feed efficiency using a regularized regression algorithm to identify immune pathways that were associated with efficiency. For statistical analysis, RNA integrity number, feed efficiency and time post LPS challenge were included in the model. At 2 h post LPS, HE pigs showed greater activation in pathways related to heat stress and creatine metabolism, but inhibition of pathways related to lysine and threonine catabolism, and immune resolving mediators (P-adj < 0.01), compared with the response in LE pigs. At 6 h post challenge there was a striking difference in cell cycle response, whereby cell cycle was activated in HE pigs but inhibited in LE pigs (P-adj < 0.01). The biggest difference in immune response was at 24h, with a strong response to LPS still evident in HE pigs, but a diminishing response in LE pigs. High efficiency pigs had activation in pathways related to oxidative stress, the metabolism of tryptophan, serine, tyrosine, threonine, glycogen, vitamin B5, and fibrin clot formation (P-adj < 0.01), which were not evident in LE pigs. The regression of pathways with feed efficiency showed that vitamin B5 metabolism was positively associated with gain:feed, whilst calcium-dependent signaling was negatively associated with gain:feed at multiple timepoints post-challenge. High efficiency pigs appeared to expand leukocyte populations at 6 h post LPS challenge, which was not evident in LE pigs. By 24 h, the response of LE pigs to LPS was minimal, but HE pigs still showed a strong response with greater utilization of energy, amino acids, and more oxidative stress. Low efficiency pigs appeared to recover faster from the LPS challenge, which was partially explained by differences in vitamin B5 utilization and calcium homeostasis. These findings confirm differences in the immune response of HE and LE pigs and provide insights related to amino acid, vitamin B5, and calcium homeostasis, which could help optimize the immune response of HE pigs. The novel analysis methods provided additional insights, which complement the original findings (T1).

Lecithin:cholesterol acyltransferase (LCAT) is the major determinant of the cholesteryl ester (CE) content of high density lipoprotein (HDL) in plasma. The selective uptake of HDL-CE is postulated to participate in delivery of tissue-derived cholesterol both to the liver and steroidogenic tissues. Recent studies comparing mice with similarly low levels of HDL, due to the absence of either of the two major HDL-associated apolipoproteins apoA-I and apoA-II, suggest that apoA-I is crucial in modulating this process, possibly through interaction with scavenger receptor class B type I (SR-BI). Because of the central role of LCAT in determining the size, lipid composition, and plasma concentration of HDL, we have created LCAT-deficient mice by gene targeting to examine the effect of LCAT deficiency on HDL structure and composition and adrenal cholesterol delivery. The HDL in the LCAT-deficient mice was reduced in its plasma concentration (92%) and CE content (96%). The HDL particles were heterogeneous in size and morphology and included numerous discoidal particles, mimicking those observed in LCAT-deficient humans. The adrenals of the male Lcat (-/-) mice were severely depleted of lipid stores, which was associated with a 2-fold up-regulation of the adrenal SR-BI mRNA. These studies demonstrate that LCAT deficiency, similar to apoA-I deficiency, is associated with a marked decrease in adrenal cholesterol delivery and supports the hypothesis that adrenal SR-BI expression is regulated by the adrenal cholesterol.

Neutrophils and B-cells in blood and head kidney of Atlantic salmon ( Salmo salar L.) challenged with infectious pancreatic necrosis virus (IPNV)

The aim of our study was to examine the effects of an immunomodulatory feed additive (OmniGen® PRO, Phibro Animal Health, Teaneck, NJ) on ruminal fermentation homeostasis and biomarkers of stress and inflammation in dairy heifers following subacute ruminal acidosis (SARA). Holstein heifers (n = 32, 8.4 ± 0.3 months old) were allocated to receive two treatments based on body weight (BW). Treatments were (1) control (CON; n = 16, no additive) and (2) OmniGen® PRO (OGPRO, n = 16, 10 g/100 kg BW, top-dressed). From d 1 to 69, and d 72 to 77, heifers were fed ad libitum a basal TMR formulated for early lactation. On d 70, the TMR offerings were reduced by 50% based on the intake of the previous 3 days. On d 71, heifers were fed ad libitum a starch challenge diet, which was a 50/50 mix steam-rolled barley and the basal TMR on a DM basis, to induce SARA. Rumen fluid and blood samples were collected on d 66, 71, and 73. Prior to the challenge, DMI, ADG, and feed efficiency were not affected by treatment. Following the challenge, all heifers experienced a 43% decline in DMI. Rumen pH was lower on the challenge day than pre and post challenge but was unaffected by treatment. Ruminal lactate was negligible pre and post challenge but increased on the challenge day; OGPRO reduced ruminal lactate compared to CON. At all sampling points, rumen total VFA were higher in OGPRO than in CON. The challenge caused fluctuations in the acetate to propionate ratio in CON, while OGPRO heifers had less variation. Two days post challenge, OGPRO heifers tended to have lower plasma cortisol, haptoglobin, and non-esterified fatty acids (NEFA) than CON heifers. The OGPRO heifers maintained the mean corpuscular hemoglobin concentration (MCHC) and platelet concentration after the challenge, while their levels declined in CON. In this study, supplementing OGPRO to heifers fed an early lactation diet improved rumen fermentation measures prior to the starch challenge and reduced the negative effects of the challenge on rumen fermentation. In addition, following the challenge, indicators of systemic inflammation tended to be lower in heifers supplemented with OGPRO.

Saffron and its constituents are reported to have a wide range of biological activities. To determine the hypoglycemic effect of hydromethanolic extract of saffron, which has rarely been dealt with in previous studies and hypolipidemic effect of saffron extract, we investigated the effects of hydromethanolic extract of saffron on healthy male rats. The present study also aimed at evaluating the effects of saffron extract on insulin secretory function of pancreatic β-cells. In this study, healthy male rats (n = 30) were divided into 3 groups of ten: The test group (saffron group), the sham group (physiologic serum group) and the normal group. The test group received 50 mg/kg saffron extract through intrap eritonal injection. The sham group was also intraperitoneally injected by 50 mg/kg of physiologic serum solvent for 2 weeks. Daily injection was repeated at 10 am. Blood samples were obtained from retro-orbital sinus before administration and on the 7th and 14th days of administration. The serum glucose and cholesterol levels were measured by the enzymatic method and insulin levels were measured using insulin kit by ELISA method. The results showed that hydromethanolic extract of saffron on the 7th day of administration, significantly decreased serum glucose in saffron group without having any effects on serum cholesterol and insulin levels. On the 14th day of administration, we found significant decrease of serum glucose and cholesterol levels in saffron group. In addition, after 2 weeks, serum insulin in test group significantly increased compared to the sham and normal groups. No changes were observed on serum glucose, cholesterol, and insulin levels in sham and normal groups on the 7th and 14th days of administration. These results suggest that saffron extract has hypoglycemic and hypolipidemic effects on healthy male rats. Moreover, saffron extract could increase their insulin secretion from pancreatic β-cells. Therefore, it should be considered in future therapeutic researches. Key words: Saffron, insulin, cholesterol, glucose, healthy male rat.

Concomitant lipopolysaccharide-induced transfer of blood-derived components including immunoglobulins into milk

We conducted a genome-wide scan using variance components linkage analysis to localize quantitative-trait loci (QTLs) influencing triglyceride (TG), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol, and total cholesterol (TC) levels in 3,071 subjects from 459 families with atherogenic dyslipidemia. The most significant evidence for linkage to TG levels was found in a subset of Turkish families at 11q22 [logarithm of the odds ratio (LOD)=3.34] and at 17q12 (LOD=3.44). We performed sequential oligogenic linkage analysis to examine whether multiple QTLs jointly influence TG levels in the Turkish families. These analyses revealed loci at 20q13 that showed strong epistatic effects with 11q22 (conditional LOD=3.15) and at 7q36 that showed strong epistatic effects with 17q12 (conditional LOD=3.21). We also found linkage on the 8p21 region for TG in the entire group of families (LOD=3.08). For HDL-C levels, evidence of linkage was identified on chromosome 15 in the Turkish families (LOD=3.05) and on chromosome 5 in the entire group of families (LOD=2.83). Linkage to QTLs for TC was found at 8p23 in the entire group of families (LOD=4.05) and at 5q13 in a subset of Turkish and Mediterranean families (LOD=3.72). These QTLs provide important clues for the further investigation of genes responsible for these complex lipid phenotypes. These data also indicate that a large proportion of the variance of TG levels in the Turkish population is explained by the interaction of multiple genetic loci.

Individuals with a heterozygous mutation at the ataxia-telangiectasia mutated gene (ATM) have been reported to be predisposed to ischemic heart disease. This report examined for the first time the effect of a heterozygous ATM mutation (ATM(+)(/-)) on plasma lipid levels and atherosclerosis intensity using ATM(+/-), ATM(+)(/+) (wild type), ATM(+)(/+)/LDLR(-)(/-) (low density lipoprotein receptor knockout), ATM(+)(/-)/LDLR(-)(/-), ATM(+)(/+)/ApoE(-)(/-) (apolipoprotein E knockout), and ATM(+)(/-)/ApoE(-)(/-) mice. Our data demonstrated that the plasma cholesterol and triglyceride levels in ATM(+)(/-) and ATM(+)(/-)/LDLR(-)(/-) mice were approximately the same as those in ATM(+)(/+) and ATM(+)(/+)/LDLR(-)(/-) control mice, respectively. In contrast, the plasma cholesterol level was significantly higher in ATM(+)(/-)/ApoE(-)(/-) mice than in ATM(+)(/+)/ApoE(-)(/-) control mice. In addition, the ATM(+)(/-)/ApoE(-)(/-) mice showed higher plasma apoB-48 levels, slower clearance for plasma apoB-48-carrying lipoproteins, and more advanced atherosclerotic lesions in the aorta compared with the ATM(+)(/+)/ApoE(-)(/-) mice. These novel results suggest that the product of ATM is involved in an apoE-independent pathway for catabolism of apoB-48-carrying remnants; therefore, superimposition of a heterozygous ATM mutation onto an ApoE deficiency background reduces the clearance of apoB-48-carrying lipoproteins from the blood circulation and promotes the formation of atherosclerosis.

In an effort to discover new mouse models of cardiovascular disease using N-ethyl-N-nitrosourea (ENU) mutagenesis followed by high-throughput phenotyping, we have identified a new mouse mutation, C699Y, in the LDL receptor (Ldlr), named wicked high cholesterol (WHC). When WHC was compared with the widely used Ldlr knockout (KO) mouse, notable phenotypic differences between strains were observed, such as accelerated atherosclerotic lesion formation and reduced hepatosteatosis in the ENU mutant after a short exposure to an atherogenic diet. This loss-of-function mouse model carries a single base mutation in the Ldlr gene on an otherwise pure C57BL/6J (B6) genetic background, making it a useful new tool for understanding the pathophysiology of atherosclerosis and for evaluating additional genetic modifiers regulating hyperlipidemia and atherogenesis. Further investigation of genomic differences between the ENU mutant and KO strains may reveal previously unappreciated sequence functionality.

Evidence exists that increased levels of physical activity decrease the population burden of cardiovascular disease (CVD). Although risk factors for CVD, including plasma lipids and lipoproteins, have been associated with physical activity, studies including a sizeable number of minority participants are lacking. Our purpose was to interrogate the longitudinal effect of physical activity on plasma lipids and lipoproteins in the African American and white participants of the Atherosclerosis Risk in Communities (ARIC) Study. Nine years of follow-up data on 8,764 individuals aged 45-64 years at baseline were used in linear mixed-effects models to estimate the association between increases in baseline physical activity on mean change in HDL, LDL, total cholesterol, and triglyceride levels. Increases in the level of activity were associated with increases in HDL in all strata and decreases in triglycerides among white participants. Physical activity was associated with LDL in all women, while the association with total cholesterol was limited to African American women. This study is one of the few to investigate the effect of physical activity on lipids and lipoproteins in a race- and sex-specific manner. Overall our results highlight the importance of physical activity on plasma lipid profiles and provide evidence for novel differential associations.

The present study was undertaken to investigate the effect of cholesterol-enriched casein (CAS) and blue lupin seed (BL) diets on the cholesterol metabolism of intact (INT) and ileorectal anastomosed (IRA) pigs. For 3 weeks, four groups of six pigs were allocated to the treatments (CAS-INT, CAS-IRA, BL-INT, and BL-IRA). Diet-induced hypercholesterolemia was inhibited by the BL through a substantial decrease in plasma LDL-cholesterol. The BL also reduced liver esterified and total cholesterol, increased hepatic LDL receptor synthesis and HMG-CoA reductase activity, and stimulated intestinal bile acid reabsorption. The neutral sterol output was higher in BL- than in CAS-fed pigs. The bile acid output was lower in IRA than in INT pigs. Surgery also prevented steroid microbial transformation, but it did not influence plasma cholesterol levels. These results suggest that the hypocholesterolemic effect of the BL, compared with the CAS, is attributable to impaired intestinal cholesterol absorption, probably involving increased bile acid reabsorption and higher contents of dietary phytosterols, both factors that reduce the micellar solubilization of cholesterol. Furthermore, according to our data, the contribution of the large intestine to cholesterol metabolism is very weak.

Effect of a Contraceptive Patch vs. Placebo (PBO) on Serum Lipid Profile

Protection and immune response in pigs intradermally vaccinated against porcine reproductive and respiratory syndrome (PRRS) and subsequently exposed to a heterologous European (Italian cluster) field strain