Promising xenograft animal model recapitulating the features of human pancreatic cancer

BACKGROUND Multiple sites of metastasis and desmoplastic reactions in the stroma are key features of human pancreatic cancer(PC).There are currently no simple and reliable animal models that can mimic these features for accurate disease modeling.AIM To create a new xenograft animal model that can faithfully recapitulate the features of human PC.METHODS Interleukin 2 receptor subunit gamma(IL2RG)gene knockout Syrian hamster was created and characterized.A panel of human PC cell lines were transplanted into IL2RG knockout Syrian hamsters and severe immune-deficient mice subcutaneously or orthotopically.Tumor growth,local invasion,remote organ metastasis,histopathology,and molecular alterations of tumor cells and stroma were compared over time.RESULTS The Syrian hamster with IL2RG gene knockout(named ZZU001)demonstrated an immune-deficient phenotype and function.ZZU001 hamsters faithfully recapitulated most features of human PC,in particular,they developed metastasis at multiple sites.PC tissues derived from ZZU001 hamsters displayed desmoplastic reactions in the stroma and epithelial to mesenchymal transition phenotypes,whereas PC tissues derived from immune-deficient mice did not present such features.CONCLUSION ZZU001 hamsters engrafted with human PC cells are a superior animal model compared to immune-deficient mice.ZZU001 hamsters can be a valuable animal model for better understanding the molecular mechanism of tumorigenesis and metastasis and the evaluation of new drugs targeting human PC.

Large animal ischemic stroke models: replicating human stroke pathophysiology

The high morbidity and mortality rate of ischemic stroke in humans has led to the development of numerous animal models that replicate human stroke to further understand the underlying pathophysiology and to explore potential therapeutic interventions.Although promising therapeutics have been identified using these animal models,with most undergoing significant testing in rodent models,the vast majority of these interventions have failed in human clinical trials.This failure of preclinical translation highlights the critical need for better therapeutic assessment in more clinically relevant ischemic stroke animal models.Large animal models such as non-human primates,sheep,pigs,and dogs are likely more predictive of human responses and outcomes due to brain anatomy and physiology that are more similar to humans-potentially making large animal testing a key step in the stroke therapy translational pipeline.The objective of this review is to highlight key characteristics that potentially make these gyrencephalic,large animal ischemic stroke models more predictive by comparing pathophysiological responses,tissue-level changes,and model limitations.

Identification of predictive MRI and functional biomarkers in a pediatric piglet traumatic brain injury model

Traumatic brain injury(TBI) at a young age can lead to the development of long-term functional impairments. Severity of injury is well demonstrated to have a strong influence on the extent of functional impairments;however, identification of specific magnetic resonance imaging(MRI) biomarkers that are most reflective of injury severity and functional prognosis remain elusive. Therefore, the objective of this study was to utilize advanced statistical approaches to identify clinically relevant MRI biomarkers and predict functional outcomes using MRI metrics in a translational large animal piglet TBI model. TBI was induced via controlled cortical impact and multiparametric MRI was performed at 24 hours and 12 weeks post-TBI using T1-weighted, T2-weighted, T2-weighted fluid attenuated inversion recovery, diffusion-weighted imaging, and diffusion tensor imaging. Changes in spatiotemporal gait parameters were also assessed using an automated gait mat at 24 hours and 12 weeks post-TBI. Principal component analysis was performed to determine the MRI metrics and spatiotemporal gait parameters that explain the largest sources of variation within the datasets. We found that linear combinations of lesion size and midline shift acquired using T2-weighted imaging explained most of the variability of the data at both 24 hours and 12 weeks post-TBI. In addition, linear combinations of velocity, cadence, and stride length were found to explain most of the gait data variability at 24 hours and 12 weeks post-TBI. Linear regression analysis was performed to determine if MRI metrics are predictive of changes in gait. We found that both lesion size and midline shift are significantly correlated with decreases in stride and step length. These results from this study provide an important first step at identifying relevant MRI and functional biomarkers that are predictive of functional outcomes in a clinically relevant piglet TBI model. This study was approved by the University of Georgia Institutional Animal Care and Use Committee(AUP: A2015 11-001) on December 22, 2015.

Maternal supply of methionine during late-pregnancy enhances rate of Holstein calf development in utero and postnatal growth to a greater extent than colostrum source

Background: Pregnancy and early life are critical periods of plasticity during which the fetus and neonate may be influenced by environmental factors such as nutrition.Maternal methionine(Met) supply in non-ruminants during pregnancy can affect offspring development and growth.Thus,the objective of this study was to investigate if increasing Met supply during late-pregnancy affects developmental parameters of the calf at birth and if either maternal Met or colostrum from Met-fed cows alters calf growth.Calves born to Holstein cows individually-fed a basal control [CON; 1.47 Mcal/kg dry matter(DM) and 15.3% crude protein] diet with no added Met or CON plus ethylcellulose rumen-protected Met(MET; Mepron? at 0.09% of diet DM; Evonik Nutrition & Care GmbH,Germany)during the last 28 ± 2 d of pregnancy were used.A total of 39 calves were in CON(n = 22 bulls,17 heifers) and 42 in MET(n = 20 bulls,22 heifers).At birth,calves were randomly allocated considering dam treatment and colostrum as fol ows: 1) calves from CON cows and colostrum from CON cows(n = 21); 2) calves from CON cows and colostrum from MET cows(n = 18); 3) calves from MET cows and colostrum from MET cows(n = 22); and 4) calves from MET cows and colostrum from CON cows(n = 20).Al calves were housed,managed,and fed individual y during the first 9 wk of life.Results: Despite greater daily DM intake pre-partum in cows fed MET(15.7 vs.14.4 ± 0.12 kg/d,P < 0.05),colostrum quality and quantity were not affected by maternal diet.At birth,MET calves had greater(P ≤ 0.05) body weight(BW,44.1 vs.42.1 ± 0.70 kg),hip height(HH,81.3 vs.79.6 ± 0.53 cm) and wither height(WH,77.8 vs.75.9 ± 0.47 cm).In contrast,concentrations of His,Lys,and Asn in plasma were lower(P ≤ 0.05) in MET calves.Regardless of colostrum source,the greater BW,HH,and WH in MET calves at birth persisted through 9 wk of age resulting in average responses of + 3.1 kg BW,+ 1.9 cm HH,and + 1.8 cm WH compared with CON.Average daily gain during the 9 wk was(P < 0.05) 0.72 ± 0.02 kg/d in MET compared with 0.67 ± 0.02 kg/d in CON calves.Respiratory scores were normal and did not differ(P > 0.05) due to maternal Met supply or colostrum source.However,fecal scores tended to be lower(P ≤ 0.10) in MET calves regardless of colostrum source.Conclusions: Increasing the maternal supply of MET during late-pregnancy enhanced growth in utero as wel as during the pre-weaning and early post-weaning periods.Although the ～ 1 kg/d greater DM intake during the last 2–3 wk prior to parturition could explain a portion of the 2 kg extra body mass of MET calves at birth,other mechanisms potential y encompassing nutrient assimilation efficiency likely played a role.Assessing the exact mechanisms sensitive to supply of Met or total amino acid supply during the latter stages of growth in utero merit further research.

The market for amino acids: understanding supply and demand of substrate for more efficient milk protein synthesis

For dairy production systems, nitrogen is an expensive nutrient and potentially harmful waste product. With three quarters of fed nitrogen ending up in the manure, significant research efforts have focused on understanding and mitigating lactating dairy cows' nitrogen losses. Recent changes proposed to the Nutrient Requirement System for Dairy Cattle in the US include variable efficiencies of absorbed essential AA for milk protein production. This first separation from a purely substrate-based system, standing on the old limiting AA theory, recognizes the ability of the cow to alter the metabolism of AA. In this review we summarize a compelling amount of evidence suggesting that AA requirements for milk protein synthesis are based on a demand-driven system. Milk protein synthesis is governed at mammary level by a set of transduction pathways, including the mechanistic target of rapamycin complex 1(mTORC1), the integrated stress response(ISR), and the unfolded protein response(UPR). In tight coordination, these pathways not only control the rate of milk protein synthesis, setting the demand for AA, but also manipulate cellular AA transport and even blood flow to the mammary glands, securing the supply of those needed nutrients. These transduction pathways, specifically mTORC1, sense specific AA, as well as other physiological signals, including insulin, the canonical indicator of energy status. Insulin plays a key role on mTORC1 signaling, controlling its activation, once AA have determined mTORC1 localization to the lysosomal membrane.Based on this molecular model, AA and insulin signals need to be tightly coordinated to maximize milk protein synthesis rate. The evidence in lactating dairy cows supports this model, in which insulin and glucogenic energy potentiate the effect of AA on milk protein synthesis. Incorporating the effect of specific signaling AA and the differential role of energy sources on utilization of absorbed AA for milk protein synthesis seems like the evident following step in nutrient requirement systems to further improve N efficiency in lactating dairy cow rations.

Nanotechnology-based approach for safer enrichment of semen with best spermatozoa

Background: Advances in nanotechnology have permitted molecular-based targeting of cells through safe and biocompatible magnetic nanoparticles(MNP). Their use to detect and remove damaged spermatozoa from semen doses could be of great interest. Here, MNP were synthesized and tested for their ability to target apoptotic(annexin V) and acrosome-reacted(lectin) boar spermatozoa, for high-throughout retrieval in a magnetic field(nanoselection). The potential impacts of nanoselection on sperm functions and performance of offspring sired by sperm subjected to nanoselection were determined. Fresh harvested and extended boar semen was mixed with various amounts(0, 87.5, and 175 μg) of MNP-conjugates(Annexin V-MNP or Lectin-MNP) and incubated(10 to15 min) for 37 °C in Exp. 1. In Exp. 2, extended semen was mixed with optimal concentrations of MNP-conjugates and incubated(0, 30, 90, or 120 min). In Exp. 3, the synergistic effects of both MNP-conjugates(87.5 μg– 30 min)on spermatozoa was evaluated, followed by sperm fertility assessments through pregnancy of inseminated gilts and performance of neonatal offspring. Sperm motion, viability, and morphology characteristics were evaluated in all experiments.Results: Transmission electron microscopy, atomic force microscopy, and hyperspectral imaging techniques were used to confirm attachment of MNP-conjugates to damaged spermatozoa. The motility of nanoselected spermatozoa was improved(P < 0.05). The viability of boar sperm, as assessed by the abundance of reactive oxygen species and the integrity of the acrosome, plasma membrane, and mitochondrial membrane was not different between nanoselected and control spermatozoa. The fertility of gilts inseminated with control or nanoselected spermatozoa, as well as growth and health of their offspring were not different between(P > 0.05).Conclusions: The findings revealed the benefit of magnetic nanoselection for high-throughput targeting of damaged sperm, for removal and rapid and effortless enrichment of semen doses with highly motile, viable,and fertile spermatozoa. Therefore, magnetic nanoselection for removal of abnormal spermatozoa from semen is a promising tool for improving fertility of males, particularly during periods, such as heat stress during the summer months.

Effects of Sodium Citrate plus Sodium Diacetate and Buffered Vinegar on Quality Attributes of Enhanced Beef Top Rounds

The effect of two antimicrobials in the enhancement solution was investigated in this study. Sensory and shelf-life characteristics were measured for beef top rounds enhanced to 110% with 0.5% sodium chloride and 0.4% sodium tripolyphosphate (CNT);CNT with a 1% solution of 80% sodium citrate plus 20% sodium diacetate (SC + D);or CNT with 2% buffered vinegar (VIN) in the final product. SC + D and VIN had less (P 0.05). However, SC + D had lower (P 0.05) for all enhancement solutions within each day. After 7 days of retail display, SC + D retarded (P < 0.05) lipid oxidation to a greater extent than VIN or CNT samples. Sodium citrate plus sodium diacetate or buffered vinegar may be included in enhancement solutions without having a large influence on subjective color, and may aid in improving sensory tenderness.

Effect of Molasses Addition at Ensiling on Ruminal In Situ Dry Matter and Nutrient Degradation of Whole-Plant Soybean Silage Harvested at Different Phenological Stages

Whole-plant soybean ensiling has limitations and challenges that affect silage fermentation patterns and reduce ruminal nutrient degradation. Perhaps either the addition of molasses at ensiling or harvesting at different phenological stages has the potential to enhance whole-plant soybean silage (WSS) ruminal degradation. This experiment was a completely randomized design with a 3 × 2 factorial arrangement of treatments evaluating the effects of molasses (0 and 40 g/kg fresh forage) and phenological stage (R5: beginning seed, R6: full seed, and R7: beginning maturity) on ruminal in situ degradation. Molasses increased effective ruminal degradability (ERD) of dry matter (DM) regardless of the phenological stage and increased ERD of crude protein at R5 and R6. The addition of molasses at later phenological stages increased ERD of neutral detergent fiber (NDF). There was a molasses × phenological stage interaction effect with greater fractions A and B of NDF at R7 and R5, respectively, but lower undegraded NDF at R5 and R6 with molasses addition. Although the benefits of adding molasses were more pronounced in R5 and R6, adding molasses to whole-plant soybean at ensiling is recommended regardless of phenological stage. Increasing the proportion of digestible nutrients at harvesting either by harvesting at later phenological stages or the addition of molasses is a powerful strategy to manage ruminal degradation of DM and nutrients in WSS.

The Fate of Aflatoxin in Corn Fermentation

A lab-scale ethanol fermentation was investigated to determine where aflatoxin concentrated during each phase of production. Four corn samples with high levels of aflatoxin (ranging from 7750 – 17,208 parts per billion) and their replicates were compared with a replicated negative control. Fractions were taken from the fermented mash, distilled ethanol, stillage, and dried corn solids (DCS). These fractions were analyzed using two different immunoassay methods and liquid chromatography tandem mass spectrometry (LC-MS/MS). Results indicated no aflatoxin was found in the distilled ethanol. Some aflatoxin (13%) was detected in the stillage, but most of the toxin was recovered in the DCSs ranging from 31% to 58%. A second series of experiments were conducted to investigate the effect of binders on dried distillers grains (DDGs). A brewers dried yeast anti-caking binder that contains glucomannon (MTB-100?), was mixed with contaminated DDGs. Addition of the binder showed a significant reduction in aflatoxin levels in comparison to a positive control. Aflatoxin binding at 2% binder w/w reached 72.5% and showed a minimal binding percentage increase of 80% at 6% binder w/w. Testing was also conducted to determine if environmental variables such as pH and temperature had any effect on the binding capabilities. Temperature near 0?C resulted in binding at 19.7% at a pH range of 6 to 8. Additionally, at a temperature of 40?C resulted in binding of 36%, 47%, and 45% at pHs 6, 7, and 8, respectively. These findings suggest that the addition of sorbents may be an effective way of salvaging contaminated DDGs.

Pharmacologic inhibition of mTORC1 mimics dietary protein restriction in a mouse model of lactation

Background:Understanding the mechanisms of N utilization for lactation can lead to improved requirement estimates and increased efficiency,which modern dairy diets currently fail to maximize.The mechanistic target of rapamycin complex 1(mTORC1)is a central hub of translation regulation,processing extra-and intra-cellular signals of nutrient availability and physiological state,such as amino acids and energy.We hypothesized that dietary amino acids regulate lactation through mTORC1,such that inhibition of mTORC1 will lead to decreased lactation performance when amino acids are not limiting.Our objectives were to assess lactation performance in lactating mice undergoing dietary and pharmacologic interventions designed to alter mTORC1 activity.Methods:First lactation mice(N=18;n=6/treatment)were fed an adequate protein diet(18%crude protein),or an isocaloric protein-restricted diet(9%crude protein)from the day after parturition until lactation day 13.A third group of mice was fed an adequate protein diet and treated with the mTORC1 inhibitor rapamycin(4 mg/kg every other day)intraperitoneally,with the first two groups treated with vehicle as control.Dams and pups were weighed daily,and feed intake was recorded every other day.Milk production was measured every other day beginning on lactation day 4 by the weigh-suckle-weigh method.Tissues were collected after fasting and refeeding.Results:Milk production and pup weight were similarly decreased by both protein restriction and rapamycin treatment,with final production at 50%of control(P=0.008)and final pup weight at 85%of control(P<0.001).Mammary phosphorylation of mTORC1’s downstream targets were decreased by protein restriction and rapamycin treatment(P<0.05),while very little effect was observed in the liver of rapamycin treated mice,and none by protein restriction.Conclusions:Overall,sufficient supply of dietary amino acids was unable to maintain lactation performance status in mice with pharmacologically reduced mammary mTORC1 activity,as evidenced by diminished pup growth and milk production,supporting the concept that mTORC1 activation rather than substrate supply is the primary route by which amino acids regulate synthesis of milk components.

No effect of exogenous melatonin on development of cryopreserved metaphase Ⅱ oocytes in mouse

Background： This study was conducted to investigate effect of exogenous melatonin on the development of mouse mature oocytes after cryopreservation. Results： First, mouse metaphase II （MII） oocytes were vitrified in the open-pulled straws （OPS）. After warming, they were cultured for 1 h in M2 medium containing melatonin at different concentrations （0, 10-9, 10-7, 10-s, 10 3 mol/L）. Then the oocytes were used to detect reactive oxygen species （ROS） and glutathione （GSH） levels （fluorescence microscopy）, and the developmental potential after parthenogenetic activation. The experimental results showed that the ROS level and cleavage rate in 10-3 mol/L melatonin group was significantly lower than that in melatonin-free group （control）. The GSH levels and blastocyst rates in all melatonin-treated groups were similar to that in control. Based on the above results, we detected the expression of gene Hsp9Oaal, Hsfl, Hspalb, Nrf2 and Bcl-xl with qRT-PCR in oocytes treated with 10-7, or 10-3 mol/L melatonin and untreated control. After warming and culture for 1 h, the oocytes showed higher Hsp90oal expression in 10-7 mol/L melatonin-treated group than in the control （P 〈 0.05）; the Hsfl, Hsp9Oaal and Bcl-xl expression were significantly decreased in 10-3 mol/L melatonin-treated group when compared to the control. Based on the above results and previous research, we detected the development of vitrified-warmed oocytes treated with either 10-7 or 0 mol/L melatonin by in vitro fertilization. No difference was observed between them. Conclusions： Our results indicate that the supplementation of melatonin （10-9 to 10-3 mol/L） in culture medium and incubation for 1 h did not improve the subsequent developmental potential of vitrified-warmed mouse MII oocytes, even if there were alteration in gene expression.

Proteome changes of porcine follicular fluid during follicle development

Background:Ovarian follicular fluid influences follicle and oocyte growth,but the fluctuation of its protein content during folliculogenesis has not been comprehensively analyzed.Here we used a shotgun approach and bioinformatics analyses to investigate and compare the proteomes of porcine follicular fluid(pFF)obtained from small(<4 mm),medium(4–6 mm)and large(>6–12 mm)follicles.Results:Follicular fluid samples containing highest estrogen levels were selected as non-atretic from small(SNA:26.1±15 ng/mL),medium(MNA:162±54 ng/mL),and large(LNA:290±37 ng/mL)follicles for proteomic analyses.We detected 1627,1699,and 1756 proteins in SNA,MNA,and LNA samples,respectively.Nearly 60–63%of total proteins were specific to each sample,11–13%were shared in pairwise comparisons,and 247 proteins were shared among all samples.Functional categorization indicated comparable gene ontology(GO)terms distribution per cellular component,molecular function,and biological process categories across samples;however,the ranking of highly significantly enriched GO terms per category revealed differences between samples.The patterns of proteinto-protein interactions varied throughout follicle development,and proteins such as serine protease inhibitor,clade E(SERPINE);plasminogen activator,urokinase(PLAU);and plasminogen activator,urokinase receptor(PLAUR)appeared stage-specific to SNA,MNA,and LNA,respectively.The“complement and coagulation cascades”was the common major pathway.Besides,properdin and fibulin-1 were abundant proteins that appeared absent in LNA samples.Conclusion:This study provides extensive and functional analyses of the pFF proteome changes during folliculogenesis and offers the potential for novel biomarker discovery in pFF for oocyte quality assessment.

Observations of High Nitrate Concentrations in Forage Bermudagrass during Severe to Exceptional Drought Years

With the prediction of climate change-induced increases in drought frequency and severity in the southeastern USA, it is important to better understand the risks that drought may pose to NO3 accumulation in bermudagrass [Cynodon dactylon (L.) Pers.] forage. This report offers observations of NO3 concentration in Bermudagrass forage samples submitted to the University of Georgia’s Feed and Environmental Water Lab (FEWL) during the extreme to exceptional drought of 2007, the severe drought of 2008, and the four preceding seasons when drought stress was minimal or absent. The probability (P) of a sample being at high risk for nitrate toxicosis was the greatest for the extreme to exceptional drought of 2007 (P = 0.160), slightly lower in the severe drought year of 2008 (P = 0.105), and the lowest for samples from the 2003-2006 growing seasons (P = 0.082) when drought stress was minimal or absent.

Use of Observed Genomic Information to Infer Linkage Disequilibrium between Markers and QTLs

Conducting genomic selection in admixed populations is challenging and its accuracy in this case largely depends on the persistence of linkage disequilibrium between single nucleotide polymorphisms (SNP) and quantitative trait loci (QTL). Inferring linkage disequilibrium (LD) between SNP markers and QTLs could be important in understanding the change of SNP marker effects across different breeds. Predicting the change in linkage disequilibrium between markers and QTLs across two divergent breeds was explored using information from the genotype data. Two different models (M1, M2) that differ in the definition of the explanatory variables were used to infer the level of LD between SNP markers and QTLs using all markers in the panel or windows of fixed number of markers. Three simulation scenarios were conducted using different number of SNPs and QTLs. In the first scenario, the resulting coefficient of determination (R2) was 0.65 and 0.52 using M1 and M2, respectively. In the second scenario, average R2 equaled 0.12 using all markers in the panel and 0.25 using 100 marker windows. Across the three simulation scenarios, it was clear that a significant portion of the variation in the change in LD between SNP markers and QTLs could be explained by information already available in the observed SNP marker data.

The pig as a preclinical traumatic brain injury model:current models,functional outcome measures,and translational detection strategies

Traumatic brain injury(TBI) is a major contributor of long-term disability and a leading cause of death worldwide. A series of secondary injury cascades can contribute to cell death, tissue loss, and ultimately to the development of functional impairments. However, there are currently no effective therapeutic interventions that improve brain outcomes following TBI. As a result, a number of experimental TBI models have been developed to recapitulate TBI injury mechanisms and to test the efficacy of potential therapeutics. The pig model has recently come to the forefront as the pig brain is closer in size, structure, and composition to the human brain compared to traditional rodent models, making it an ideal large animal model to study TBI pathophysiology and functional outcomes. This review will focus on the shared characteristics between humans and pigs that make them ideal for modeling TBI and will review the three most common pig TBI models–the diffuse axonal injury, the controlled cortical impact, and the fluid percussion models. It will also review current advances in functional outcome assessment measures and other non-invasive, translational TBI detection and measurement tools like biomarker analysis and magnetic resonance imaging. The use of pigs as TBI models and the continued development and improvement of translational assessment modalities have made significant contributions to unraveling the complex cascade of TBI sequela and provide an important means to study potential clinically relevant therapeutic interventions.

An integrative multivariate approach for predicting functional recovery using magnetic resonance imaging parameters in a translational pig ischemic stroke model

Magnetic resonance imaging(MRI)is a clinically relevant,real-time imaging modality that is frequently utilized to assess stroke type and severity.However,specific MRI biomarkers that can be used to predict long-term functional recovery are still a critical need.Consequently,the present study sought to examine the prognostic value of commonly utilized MRI parameters to predict functional outcomes in a porcine model of ischemic stroke.Stroke was induced via permanent middle cerebral artery occlusion.At 24 hours post-stroke,MRI analysis revealed focal ischemic lesions,decreased diffusivity,hemispheric swelling,and white matter degradation.Functional deficits including behavioral abnormalities in open field and novel object exploration as well as spatiotemporal gait impairments were observed at 4 weeks post-stroke.Gaussian graphical models identified specific MRI outputs and functional recovery variables,including white matter integrity and gait performance,that exhibited strong conditional dependencies.Canonical correlation analysis revealed a prognostic relationship between lesion volume and white matter integrity and novel object exploration and gait performance.Consequently,these analyses may also have the potential of predicting patient recovery at chronic time points as pigs and humans share many anatomical similarities(e.g.,white matter composition)that have proven to be critical in ischemic stroke pathophysiology.The study was approved by the University of Georgia(UGA)Institutional Animal Care and Use Committee(IACUC;Protocol Number:A2014-07-021-Y3-A11 and 2018-01-029-Y1-A5)on November 22,2017.

Effects of maternal dietary omega-3 polyunsaturated fatty acids and methionine during late gestation on fetal growth,DNA methylation,and mRNA relative expression of genes associated with the inflammatory response,lipid metabolism and DNA methylation in placenta and offspring’s liver in sheep

Background:Omega-3 PUFA or methionine(Met)supply during gestation alters offspring physiology.However,the effect of both nutrients on fetal development has not been explored.Our objective was to determine the effects of supplementation of these two nutrients during late gestation on fetal growth,DNA methylation,and mRNA expression of genes associated with the inflammatory response,and DNA methylation.Ewes(n=5/treatment)were fed from day 100 to 145 of gestation one of the following treatments:1)basal diet(NS)without fatty acids(FS)or methionine(MS)supplementation;2)FS(10 g/kg Ca salts,source omega-3 PUFA);3)MS(1 g/kg rumen protected methionine);and 4)FS and MS(FS-MS).On day 145,ewes were euthanized,and data from dams and fetus was recorded.Placenta(cotyledon),fetal liver,and blood samples were collected.Results:A treatments interaction on fetal liver weight,ewe body weight and body condition score(BCS)was observed;FS-MS were heavier(P<0.01)than FS and MS,and FS-MS ewes had a better(P=0.02)BCS than NS.Methionine increased(P=0.03)ewe plasma glucose concentration.Fetal liver global DNA methylation increased(P<0.01)in FS and MS.Dietary treatments modify the mRNA relative expression on some of the genes evaluated.In the fetal liver,FS increased(P=0.04)the mRNA relative expression of arachidonate-5-lipoxygenase-activatingprotein and tended to decrease(P=0.06)methionine-adenosyltransferase-1A.Moreover,MS decreased(P=0.04)DNA-methyltransferase-1 and tended to decrease(P=0.08)free-fatty-acid-receptor-1 mRNA relative expression.Furthermore,FS-MS decreased mRNA relative expression of tumor-necrosis-factor-alpha(P=0.05),peroxisomeproliferator-activated-receptor-delta(P=0.03)and gamma(P=0.04),tended to decrease(P≤0.09)interleukin-6,fatty-acid-transport-protein-1,and delta-5-desaturase,and increased adenosylhomocysteinase(P=0.04)mRNA relative expression.In cotyledon,FS tended to decrease fatty acid binding protein 4(P=0.09)mRNA relative expression.Conclusion:Omega-3 PUFA and Met supplementation improves dam’s performance in late gestation,which was positively correlated with an increase in offspring’s liver development.Moreover,FS-MS decreased mRNA relative expression of proinflammatory cytokines,and lipogenic genes,and increased the expression on an enzyme that has an important role in methylation.

Effects of xylanase,protease,and xylo-oligosaccharides on growth performance,nutrient utilization,short chain fatty acids,and microbiota in Eimeria-challenged broiler chickens fed high fiber diet

A 21-d experiment was conducted to study the effect of xylanase,protease,and xylo-oligosaccharides on growth performance,nutrient utilization,gene expression of nutrient transporters,cecal short-chain fatty acids(SCFA),and cecal microbiota profile of broilers challenged with mixed Eimeria spp.The study utilized 392 zero-d-old male broiler chicks allocated to 8 treatments in a 4×2 factorial arrangement,as follows:corn-soybean meal diet with no enzyme(Con);Con plus xylanase alone(XYL);Con plus xylanase combined with protease(XYL+PRO);or Con plus xylo-oligosaccharides(XOS);with or without Eimeria challenge.Diets were based on a high-fiber(100 g/kg soluble fibers and 14 g/kg insoluble fibers)basal diet.At d 15,birds in challenged treatment were gavaged with a solution containing Eimeria maxima,Eimeria acervulina,and Eimeria tenella oocysts.At d 21,birds were sampled.Eimeria depressed(P<0.01)growth performance and nutrient utilization,whereas supplementation had no effect.There were significant Eimeria×supplementation interactions for the sugar transporters GLUT5(P=0.02),SGLT1(P=0.01),SGLT4(P<0.01),and peptide transporter PepT1(P<0.01)in jejunal mucosa.Eimeria challenge increased the expression of GM-CSF2(P<0.01)and IL-17(P=0.04)but decreased(P=0.03)IL-1βexpression in the cecal tonsil.Eimeria×supplementation interactions for cecal acetate,butyrate,and total SCFA showed that concentrations increased or tended to be greater in the supplemented treatments,but only in non-challenged birds.Birds challenged with Eimeria spp.had higher concentrations of isobutyrate(P<0.01),isovalerate(P<0.01),and valerate(P=0.02)in cecal content.Eimeria challenge significantly(P<0.01)decreased the microbial richness and diversity,and increased(P<0.01)the proportion of Anaerostipes butyraticus,Bifidobacterium pseudolongum,and Lactobacillus pontis.In conclusion,Eimeria infection depressed growth performance,nutrient utilization with regulating nutrient transporters.Furthermore,Eimeria challenge shifted the microbial profile and reduced microbial richness and diversity.On the other hand,enzyme supplementation showed limited benefits,which included increased concentrations of SCFA.

Differential IFN-Gamma (IFN-γ), Interleukin 10 (IL-10) and Cardiac Troponin I (cTnI) Responses in Natural Bovine Trypanosomosis in Nigeria

Trypanosomosis is major drawback to profitable livestock production in sub-Sahara African, including Nigeria. Knowledge of the cytokines production in the phase of natural infection may help to better diagnose, treat and prevent bovine trypanosomosis. The purpose of the this study was to determine the levels of interferon-gamma (IFN-γ), interleukin-10 (IL-10) and cardiac troponin–I (cTnI) in the sera of cattle naturally infected with T. brucei, T. congolense and T. vivax and correlate these levels with parasitaemia and PCV of the infected animals. Five milliliter of blood samples were collected via the jugular vein from 411 randomly selected cattle into EDTA and non-citrated bottle. PCV was determined manually using HCT. Trypansomes were detected and characterized by microscopy and PCR, respectively. Serum levels of IFN-γ, IL-10 and cTnI were determined using commercial ELISA kit. Data were summarized using descriptive statistic and significance of differences determined by ANOVA. Of the 62 samples positive for trypanosomes by microscopy, 50 samples were confirmed to species level by PCR. The sera levels of IFN-γ, IL-10 and cTnI of infected cattle were higher than non-infected cattle. The differences were not significant (p γ, IL-10 and cTnI in cattle with natural trypanosomosis. Further investigation is required to understand the specific effect of trypanosomes on myocardiac integrity and interaction between the two cytokines in natural trypanosomosis in cattle.

Total amino acid level affects the results of standardized ileal digestibility assays for feed ingredients for swine

It has been demonstrated that the fiber content of oilseed meals and total amino acids(AA)in distillers'grains products affects standardized ileal digestibility(SID)values from swine assays and that total protein concentration affects the AA digestibility in assays using chickens.This analysis was conducted to test the hypothesis that the total AA content of test samples affects the SID assay results.Databases containing total AA profiles and SID values of 20 feedstuffs commonly fed to pigs from 2 sources,AMINODat 5.0(Evonik Industries,2015)and the Nutrient Requirements of Swine,12th edition(NRC,2012)were used to compare AA concentration effects on standardized ileal digestibility coefficients.Databases were compared with AA and SID available in both data sets.The total AA values were similar for the 2 databases with an R2 of 0.979(P<0.001).The linear relationship between digestibility co-efficients from the 2 databases was highly significant,an R2 of 0.810(P<0.001).Both databases had increased SID values with increasing AA contents within and across feed ingredients.The SID=f(AA concentration)relationship was confirmed with an individual paper.Since SID,as typically measured,is a function of both digestion and absorption,both processes following MichaeliseMenten kinetics,SID=f(AA concentration)may simply be a natural phenomenon.Other reasons for the relationship were explored.Methods of estimating endogenous AA losses and misapplication statistical procedures may contribute to variation in results and at least partially explain why SID=f(AA concentration).