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Leaky gut may be caused by acidotic conditions in the hindgut and lead to systemic inflammation and negative effects on the gastrointestinal tract. The objective was to determine the effects of induced hindgut acidosis in sheep on cecal pH, ruminal fermentation, and gut permeability. Ruminally and cecally cannulated ewes [n = 11; body weight (BW) = 49 ± 4 kg] were assigned to one of two treatments: control (CON; n = 5) or induced hindgut acidosis (HGA; n = 6). To induce hindgut acidosis, 3 g wheat starch/kg BW per 24 h was continuously infused via the cecal cannula for 4 d. Control ewes received a constant infusion of deionized water. The diet contained 40% grass hay pellets, 35% whole shelled corn, 15% dried distillers grains, and 10% alfalfa cubes. Chromium EDTA was dosed once daily via the cecal cannula as a marker of gut permeability. Rumen, cecal, and fecal samples were collected to determine pH and volatile fatty acids (VFA). Rumen fluid was collected on d 4 for an ex vivo fermentation. Flasks were incubated for 24 h to determine pH, VFA, ammonia and in vitro dry matter digestibility. Tissue samples from the ileum, cecum, and colon were mounted in Ussing chambers to measure transepithelial electrical resistance (TEER). There was a treatment × time effect (P = 0.05) for cecal pH with HGA ewes having lesser cecal pH after d 1. By d 4, cecal pH had declined to 5.07 for HGA ewes compared with CON ewes which remained above 6.40 throughout the experiment. A treatment × time interaction was also observed (P < 0.01) for fecal pH and followed the same trend as cecal pH. Rumen pH was not affected (P = 0.87) by the interaction of treatment and time but was affected (P < 0.01) by treatment as ewes on the HGA treatment had a lesser rumen pH than CON ewes. Control ewes had lesser ruminal VFA and ammonia concentrations than HGA ewes (P < 0.01). Urinary Cr recovery was not affected by the interaction of treatment and time, or treatment (P ≥ 0.13). In vitro dry matter disappearance was not affected by cecal infusion treatment (P = 0.60). There was no effect (P = 0.78) of treatment on ex vivo pH. There were no effects (P ≥ 0.11) of treatment, time, or their interaction on ex vivo ammonia or total VFA concentration. In cecal tissue, TEER tended (P = 0.09) to be greater in CON ewes than HGA ewes. In contrast, TEER was not different (P ≥ 0.83) in ileal or colonic tissues between treatments. Induced hindgut acidosis in sheep altered rumen fermentation and tended to increase ex vivo measures of cecal permeability, but did not affect ex vivo rumen fermentation. [ABSTRACT FROM AUTHOR]

Periods of decreased feed intake may disrupt function of the intestinal barrier. Feeding NutriTek® (NTK; Diamond V, Cedar Rapids, IA), a postbiotic from S. cerevisiae fermentation (SCFP), improved health and supported anti-inflammatory functions. We investigated the effects of feeding NTK to cows before and during a period of feed restriction (FR) designed to model periods of intestinal barrier dysfunction. In total, 16 multiparous cows (97.1 ± 7.6 DIM; n = 8/group) were fed a control diet (CON) or CON plus 19 g/d NTK for 9 wk (Phase 1; P1) and then were subjected to an FR challenge for 5 d, during which they were fed 40% of their ad libitum intake from the 7 d prior to FR. Milk yield (MY) and DMI were collected daily. During FR, milk was collected daily for composition, blood daily to measure plasma biomarkers and to measure monocyte and neutrophil phagocytosis and oxidative burst on d 1, 3, and 5. Data were analyzed using a mixed model in SAS 9.4. All data were subjected to repeated measures ANOVA. Dietary treatment (TRT), Day, and their interaction (TRT × Day) were considered as fixed effects and cow as the random effect. For analysis of P1, data collected during a 7-d adaptation phase were used as a covariate. During P1, NTK cows tended to have greater DMI and had greater fat, ECM and FCM yields, and feed efficiency (ECM/DMI and FCM/DMI). Protein yield tended to be greater in NTK compared with CON cows. A tendency for greater monocyte phagocytosis was detected with NTK. However, during FR, feeding NTK led to lower MY and lactose yield and tended to lower solids percentage. While NTK cows tended to have reduced neutrophil oxidative burst than CON cows during FR (NTK: 26.20%, CON: 36.93%), there was no difference in phagocytosis (NTK: 7.92%, CON: 6.31%). Plasma biomarkers of energy metabolism, liver function, inflammation, and oxidative stress during the FR period did not differ. Overall, results suggested that feeding NTK increased the yield of FCM, ECM, feed efficiency and milk components prior to FR. [ABSTRACT FROM AUTHOR]

The objective of this study was to determine the effect of a dry versus a molasses-based liquid supplement on ruminal butyrate concentration, gastrointestinal tract (GIT) barrier function, inflammatory status, and performance of newly received feedlot cattle. In experiment 1, 60 mixed breed steers (234 ± 2.1 kg) were weaned, held overnight at a sale barn, then transported 14 h to Purdue University. After arrival, steers were weighed, blocked by body weight, and allotted within block to treatments (six pens per treatment and five steers per pen). Diets consisted of 45% roughage and 55% concentrate (dry matter basis). Treatments differed in the supplement source as follows: DRY: 10% dry supplement or LIQUID : 10% liquid molasses-based supplement. Feed intake, average daily gain (ADG), and gain:feed were determined for the three 21-d periods and overall. In experiment 2, 16 crossbred heifers (246 ± 7.5 kg) were used (8 heifers per treatment). Diets were the same as in experiment 1 and were fed for 60 d. On d 56 ruminal fluid samples were collected at 0, 3, 6, and 9 h after feeding. To mimic a stress event, heifers were transported for 4 h on d 61, rested overnight, and transported 12 h on d 62. Blood was collected from heifers immediately prior to transport and immediately upon their return. Gut barrier function using a Cr-EDTA marker was determined after transportation. Data were analyzed using the GLIMMIX procedure of SAS. Steers fed the liquid supplement had greater (P  ≤ 0.03) ADG through d 42 and overall compared to steers fed the dry supplement. Feed intake did not differ (P  = 0.25) between treatments from d 0 to d 21. However, steers fed the liquid supplement showed greater (P  < 0.001) dry matter intake after d 21 and overall compared to those fed the dry supplement. Steers fed the liquid supplement tended (P  < 0.09) to have reduced serum haptoglobin and lipopolysaccharide-binding protein (LBP) compared to those fed the dry supplement. Heifers fed the liquid supplement had greater (P  = 0.02) Cr in urine and tended (P  = 0.07) to have lower serum LBP after transport compared to those fed the dry supplement. Heifers fed the liquid supplement had 72% lower serum haptoglobin before, but only a 19% lower serum haptoglobin after transport compared to animals fed the dry supplement (treatment × time; P  = 0.07). Therefore, the liquid supplement altered GIT barrier function, and improved inflammatory status, resulting in increased growth of receiving cattle. [ABSTRACT FROM AUTHOR]

Stress negatively affects the gastrointestinal tract (GIT) barrier function, resulting in compromised animal health. A deeper understanding of how diet and stress impacts the GIT barrier function in feedlot cattle is needed. Aspirin decreases mucus production and mucosal repair in the GIT and could be used as a model for GIT barrier dysfunction research. The objective of this study was to evaluate the effectiveness of aspirin to induce GIT barrier dysfunction in beef cattle. In experiment 1, sixteen crossbred heifers (425.0 ± 8.6 kg) were allotted to 0, 50, 100, or 200 mg/kg body weight (BW) aspirin doses based on BW. Experiment 1 consisted of two periods separated by 4 wk where four heifers per treatment received the same aspirin dose during each period. Heifers were fed a 49.4% corn silage and 50.6% concentrate diet. The 200 mg/kg BW aspirin treatment was dosed as a 100 mg/kg BW aspirin oral bolus 36 and 24 h prior to Cr-ethylenediaminetetraacetic acid (EDTA) dosing (1 liter; 180 mM). The 50 and 100 mg/kg BW aspirin treatments were dosed as an oral bolus 24 h prior to Cr-EDTA dosing. Urine was collected every 3 h for 48 h and analyzed for Cr. Serum was collected at 0 and 48 h and analyzed for lipopolysaccharide-binding protein (LBP), interleukin-6, serum amyloid A (SAA), haptoglobin, and aspartate aminotransferase. In experiment 2, sixteen crossbred steers (576.0 ± 14.2 kg) fed a similar diet were allotted by BW to the 0 and 200 mg/kg BW aspirin treatments (eight steers/treatment) and were slaughtered 24 h after the last dose. Jejunal tissues were collected, and claudin (CLDN) 1, 2, and 3, occludin, and zonula occludens tight junction messenger ribonucleic acid (mRNA) expression was determined. Data were analyzed using the MIXED procedure of SAS. Urinary Cr excretion increased linearly at hours 3, 6, 9, and 12 (P ≤ 0.04) as aspirin dose increased from 0 to 200 mg/kg. Aspirin linearly increased Cr absorption (P = 0.02) and elimination (P = 0.04) rates and linearly decreased mean retention time of Cr (P = 0.02). Aspirin increased SAA (P = 0.04) and tended to increase LBP (P = 0.09) in serum but did not affect any other serum inflammatory marker (P ≥ 0.19). Aspirin tended to increase jejunal CLDN-1 mRNA expression (P = 0.10) but did not affect the mRNA expression of other genes regulating tight junction function (P ≥ 0.20). Results from this study indicate that aspirin disrupts the GIT barrier function in beef cattle and has a potential as a model in GIT permeability research. [ABSTRACT FROM AUTHOR]

The integrity of the intestinal barrier is crucial for regulating the passage of pathogens and toxins, while facilitating nutrient absorption. The everted gut sac technique, an ex-vivo technique, can be used to study interventions on barrier function. This cost-effective approach utilizes relatively large gut segments to study specific intestinal regions. Typically, intact (non-stripped) intestinal segments are used, but their use may underestimate permeability due to the medial positioning of blood vessels relative to the seromuscular layer and serosa. However, removing these layers risks physical damage, resulting in an overestimation of intestinal permeability. Therefore, we investigated the impact of stripping jejunal segments on permeability to fluorescein isothiocyanate-dextran (FITC, 4 kDa) and tetramethylrhodamine isothiocyanate-dextran (TRITC, 40 kDa), and on the absorption of glucose, lysine, and methionine in jejunal segments from 80 piglets at 8 d postweaning. Piglets were subjected to either high or low sanitary housing conditions and diets provoking intestinal protein fermentation or not, expected to influence intestinal permeability. Stripping of the seromuscular layer and serosa increased the passage of 4 kDa FITC-dextran (stripped vs. non-stripped; 1.1 vs. 0.9 pmol/cm2/min, P < 0.001), glucose (40.0 vs. 19.1 pmol/cm2/min, P < 0.001), lysine (2.5 vs. 2.0 nmol/cm2/min, P < 0.001), and methionine (4.1 vs. 2.7 pmol/cm2/min, P < 0.001). As permeability increased, the differences in methionine passage between stripped and non-stripped intestinal segments also increased (slope = 1.30, P = 0.009). The coefficients of variation were comparable between stripped and non-stripped intestines (over all treatments, stripped vs. non-stripped 38% vs. 40%). Stripping, by isolating mucosal processes without introducing additional variation, is thus recommended for studies on intestinal permeability or absorption., (© The Author(s) 2024. Published by Oxford University Press on behalf of the American Society of Animal Science.)

Heat stress (HS) compromises farm animal productivity and it is thus both an economic and food security issue. Reduced output (e.g., milk yield, muscle growth, egg production) during HS was traditionally thought to result from decreased nutrient intake (a classic biological response shared by all animals during environmental-induced hyperthermia). Utilizing pairfeeding experimental designs clearly indicate that reduced feed intake only accounts for a portion of the decreased productivity. The etiological epicenter of many (if not most) of the physiological consequences of HS is gastrointestinal tract hyperpermeability. For reasons not clearly understood, HS compromises the intestinal barrier allowing luminal antigens to infiltrate into local and systemic circulation. The invading antigens are recognized and stimulate the immune system. An activated immune system rearranges the hierarchy of nutrient partitioning, and the entire body participates in ensuring that leukocytes receive the fuel and building blocks necessary to mount an effective response. Central to this is increased insulin secretion coupled with simultaneous decreased adipose tissue and skeletal muscle insulin resistance. Activated leukocytes switch their energetic metabolism away from oxidative phosphorylation to aerobic glycolysis. Utilizing a LPS-Euglycemic clamp we have estimated that an intensely activated immune system can use more than 2 kg of glucose/d in lactating cows. We extended this in growing steers and pigs and the amount of glucose used is approximately 1 g/Kg BW0.75/h regardless of species and physiological state. If the heat load is severe enough, the consequences can become lethal. This is primarily explained by inflammation induced hypercoagulation, endothelial barrier dysfunction and disseminated intravascular coagulation. If the animal survives, these changes result in the reprioritization of fuel selection during HS which appears to be primarily responsible for reduced animal productivity during the warm summer months. [ABSTRACT FROM AUTHOR]

The objective of this study was to determine if feed restriction (FR) followed by feeding a high concentrate (HC) diet would induce leaky gut syndrome (LGS) in cattle. Changes in circulatory and fecal biomarkers were utilized to assess LGS. Individually penned Holstein heifers (n=16; BW=343.2 ± 3.2 kg) were used in a completely randomized design with animal as the experimental unit. Cattle were fed a forage-based diet (71.0% hay, 12.0% cracked corn, 17.0% DDGS) twice daily ad libitum from day 0-14; from day 15-17, the daily ration was restricted by 60%. One of two HC diets (n=8 each) were fed ad libitum from day 18-30: HC75=17.0% hay, 70.0% cracked corn, 13.0% DDGS; HC65=26.5% hay, 60.0% cracked corn, 13.5% DDGS. Blood and fecal samples were collected prior to a.m. feed delivery (d -1, 0, 14, 15, 18, 20, 24, 30) and analyzed for serum amyloid-A (SAA), haptoglobin (Hp), lipopolysaccharide-binding protein (LBP), fecal lactoferrin, and fecal pH. Variables were analyzed using a mixed model with repeated measures; study day, treatment and the interaction were fixed effects. Following FR, SAA was significantly increased from day 14 to 24 (27,406.20 ± 8,917.00 ng/mL; P = 0.046). Similarly, LBP was significantly increased from day 14 to 20 (353.71 ± 44.40 ng/mL; P < 0.0001). Fecal lactoferrin increased significantly from day 18 to 20 (293.9 ± 57.54 ng/g; P = 0.0001). Fecal pH decreased significantly (1.48 ± 0.09; P < 0.0001) from day 14 to 30 and from day 18 to 30 (1.57 ± 0.08; P < 0.0001) reaching its nadir on day 30 (5.35). No meaningful changes were observed for Hp. Based on these data, FR followed by feeding a HC diet effectively experimentally induced LGS as indicated by observed changes in SAA, LBP, fecal lactoferrin and fecal pH. [ABSTRACT FROM AUTHOR]

Of the many roles that the gastrointestinal tract (GIT) must facilitate, coordinated regulation of molecule movement across the gastrointestinal is key. This coordinated regulation allows for nutrient absorption while limiting permeation of non-desired molecules and is referred to as barrier function. Barrier function of the GIT includes intrinsic, extrinsic, and immunological factors, but generally permeability assessments are conducted to evaluate the whole-animal response and most studies focus on the extrinsic components of GIT barrier function. Common approaches to evaluate GIT permeability include the use of inert external markers varying in size and their measurement in blood or urine. Methodology exists for ex vivo, in vitro, and in vivo assessment; however, only in ex vivo and in vitro methods currently allow regionally specific information. Moreover, ex vivo studies have been essential to further elucidate factors that affect permeability such as osmolality, pH, and bioactive components such as histamine and lipopolysaccharides. For ruminants, extensive microbial degradation challenges the use of fermentable substrates as markers when assessing permeability and there is interest to compartmentalize permeability by region of the GIT. Recent research using dual markers to allow for total GIT and postruminal permeability have highlighted that regional permeability responses may be affected by production system, environmental conditions, and dietary characteristics. Future research is needed to improve the understanding of factors that affect GIT permeability, the regions involved, and strategies to support regulation of permeability. [ABSTRACT FROM AUTHOR]

Inflammation emanating from the gastrointestinal tract (GIT) is a key component to suboptimal farm animal productivity. Gut inflammation encompasses more than just acidosis and is a widespread condition in high producing feedlot or dairy cows which is commonly associated with feeding high grain (starch) ration. Many of the direct causes of gastrointestinal inflammation are related to the microbial fermentation in the rumen and in the hindgut, both directly and indirectly. However, many of the impacts on production are "silent" from a health perspective, and are not limited to a single compartment of the gastrointestinal tract. Gastrointestinal inflammation is often linked with tight junction dysfunction which is accompanied by gross morphological changes that reduce intestinal barrier function, allowing for intestinal luminal contents and microbes to infiltrate into portal and systemic blood (leaky gut). Lipopolysaccharide (LPS) is an endotoxin found in gram negative bacteria which is a potent antigen, which can cause systemic responses including widespread inflammation. Leukocyte antigen recognition initiates a metabolic switch from oxidative phosphorylation to aerobic glycolysis and thus the immune system becomes an obligate glucose utilizer. The reprioritization in the hierarchy of glucose trafficking reduces glucose availability to support milk and muscle synthesis. GIT hyperpermeability and the ipso facto immune stimulation is accompanied by hypophagia and a unique endocrine footprint characterized by hyperinsulinemia and hyperglucagonemia. Preventing leaky gut should be the nutritionists primary objective and multiple strategies should be considered. The use of probiotics, organic acids, and phytochemicals offer potential alternatives to antimicrobials (ATA) that can alter the microbial population and reduce: 1) microbial activity that leads to the development of conditions leading to barrier dysfunction, 2) impacts of fermentation endproducts on epithelial tissues, and 3) immunostimulation due to LPS reaching the host. [ABSTRACT FROM AUTHOR]

Physiological and environmental stresses such as the transition into lactation and heat load contribute to gastrointestinal tract (GIT) dysfunction. The nonruminant gastrointestinal tract has mechanisms to cope with pro-oxidant and pro-inflammatory stressors arising from the gut lumen or within intestinal cells. One-carbon metabolism (OCM) contributes to anti-oxidant capacity via the production of glutathione (GSH) and taurine, and the synthesis of phospholipid, creatine, and the osmolyte glycinebetaine among others. A multipronged approach was used to assess the biological relevance of OCM and closely-related pathways on GIT function in dairy cows. Ruminal papillae (Rum) and scrapings from duodenum (Duo), jejunum (Jej), and ileum (Ile) were collected at slaughter from eight multiparous Holstein cows averaging 128 ± 12 d in milk and producing 39 ± 5 kg/d. A MIXED model ANOVA with preplanned orthogonal contrasts was used for statistical analysis. Methionine adenosyl transferase 1 activity (MAT) was ~10-fold greater (P < 0.01) and cystathionine β-synthase activity doubled in Rum vs. small intestine. Total glutathione peroxidase (GPX) activity was greatest (P = 0.03) in Ile, but similar to Rum. Activity and mRNA abundance of betaine-homocysteine S-methyltransferase were undetectable. There was a 2.5-fold greater protein abundance of GPX1 (P < 0.01) and a ~2-fold greater abundance of GPX3 (P < 0.01) in Rum vs. small intestine. Among the various amino acids (AA) with roles in OCM or closely-related pathways (e.g. creatine synthesis), concentrations of arginine, aspartate, glutamine, methionine, and serine were lower (P < 0.01) in Rum vs. small intestine. Unlike AA, concentrations of OCM-related intermediates S-5'-adenosyl-homocysteine (SAH), glycinebetaine, carnitine, creatine (CRE), and cysteinesulfinic acid were greater (P < 0.01) while taurine was lower in Rum vs. small intestine. Intermediates of the folate cycle were undetectable. The fact that S-adenosylmethionine (SAM) was undetectable while MAT activity and SAH were greater in Rum suggested that availability of SAM (a methyl donor) is a key determinant of flux through the folate and methionine cycles in the GIT. Except for adenosine, concentrations of glutamate, glycine, α-ketoglutarate, hypotaurine, and GSH were lowest in Ile. Together, the data underscored unique differences in activity of one-carbon metabolism and related pathways across sections of the GIT., (© The Author(s) 2023. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The negative impacts of stress on gastrointestinal tract (GIT) barrier function can result in compromised animal growth and health. Aspirin is known to cause mucosal injury leading to increased gut permeability and tight junction damage and can be used as a model to study leaky gut in cattle. The objective of this study was to determine the long-term impact of aspirin-induced chronic leaky gut on cattle growth and carcass attributes. Two treatments were evaluated in two studies: control (no aspirin) or 0.25% of the diet dry matter (DM) aspirin fed daily. Diets consisted of 50% corn, 24% dried distillers grains, 20% corn silage, and 6% supplement on a DM basis. In experiment 1, sixteen Angus × Simmental heifers, allotted by body weight (BW) and breed composition, were fed diets for 154 d. On day 155, heifers were dosed with 1 liter of a 180-mM Cr-ethylenediaminetetraacetic acid solution using an esophageal tube and had urine collected every 1.5 to 3 h for 48 h for analysis of Cr as a measure of gut leakiness. In experiment 2, ninety-six Simmental × Angus steers (355.0 ± 14.8 kg) were allotted by BW and breed composition and fed treatment diets for 159 d. Weight was recorded monthly and serum was collected on day 159 and analyzed for lipopolysaccharide-binding protein (LBP), interleukin-6 (IL-6), serum amyloid A (SAA), haptoglobin, and aspartate aminotransferase (AST). Data were analyzed using the MIXED procedure of SAS. Heifers fed 0.25% aspirin in experiment 1 excreted more Cr into urine compared with heifers not fed aspirin (overall treatment effect, P = 0.01). In experiment 2, aspirin tended to increase serum LBP (P = 0.06) but had no effect on concentrations of IL-6, haptoglobin, SAA, or AST (P ≥ 0.25). Aspirin tended to decrease average daily gain (P = 0.10), decreased hot carcass weight and rib-eye area (P ≤ 0.05), and increased fat thickness, marbling score, and yield grade (P ≤ 0.02). Aspirin tended to increase kidney, pelvic, and heart fat percentage (P = 0.10) and had no effect on liver abscesses (P ≥ 0.80). This study indicates that leaky gut induced by long-term administration of aspirin has negative impacts on feedlot performance and carcass composition. The negative impact of aspirin-induced leaky gut on animal performance suggests that chronic leaky gut caused by other factors (subacute acidosis, stress) may be a significant problem for the feedlot industry., (© The Author(s) 2021. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)