Fat composition of milk replacer influences postprandial and oxidative metabolisms in dairy calves fed twice daily.

Milk replacers (MR) for calves contain alternative fat sources as substitute for milk fat. This substitution leads to differences in fat properties, such as the fatty acid profile and the triglyceride structure. This study evaluated how fat composition in MR affects gastrointestinal health, blood redox parameters, and postprandial metabolism in calves fed twice daily. Forty-five individually housed male Holstein-Friesian calves (2.3 ± 0.85 d of age) were assigned to 1 of 15 blocks based on the age and the day of arrival. Within each block, calves were randomly assigned to 1 of 3 experimental diets and received their respective diet from arrival until 35 d after arrival. The 3 experimental diets (n = 15 per treatment group) consisted of an MR with a blend of vegetable fats containing rapeseed and coconut (VG), an MR with only animal fats from lard and dairy cream (AN), and an MR containing a mixture of animal and vegetable fats including lard and coconut (MX). The fatty acid profile of each MR was formulated to resemble that of bovine milk fat while using only 2 fat sources. All MR were isoenergetic, with 30% fat (% DM), 24% crude protein, and 36% lactose. Chopped straw and water were available ad libitum from arrival onward but no starter feed was provided. Daily milk allowances were 6.0 L from d 1 to 5, 7.0 L from d 6 to 9, and 8.0 L from d 10 to 35, divided into 2 equal meals and prepared at 135 g/L (13.5% solids). Fecal appearance was scored daily; calves were weighed and blood was drawn on arrival and weekly thereafter. Urine and feces were collected over a 24-h period at wk 3 and 5 to determine apparent total-tract digestibility and assess gastrointestinal permeability using indigestible markers. Postprandial metabolism was evaluated at wk 4 by sequential blood sampling over 7.5 h, and the abomasal emptying rate was determined by acetaminophen appearance in blood. Fat composition in MR did not affect growth, MR intake, gastrointestinal permeability, nor nutrient digestibility. The percentage of calves with abnormal fecal scores was lower at wk 2 after arrival in calves fed VG than MX, whereas AN did not differ from the other treatments. Calves fed AN and MX had higher thiobarbituric acid reactive substances measured in serum than VG, whereas plasma ferric-reducing ability was greater in calves fed MX than VG. Postprandial acetaminophen concentrations did not differ across treatment groups, but the area under the curve was smaller in calves fed VG than in the other 2 treatments, which is indicative of a slower abomasal emptying. Postprandial serum triglyceride concentration was greater in calves fed AN than VG, whereas MX did not differ from the other treatments. Based on these outcomes, all 3 fat blends can be considered suitable for inclusion in MR for calves.

Circulating markers of intestinal barrier injury and inflammation following exertion in hypobaric hypoxia.

Hypoxia induced intestinal barrier injury, microbial translocation, and local/systemic inflammation may contribute to high-altitude associated gastrointestinal complications or symptoms of acute mountain sickness (AMS). Therefore, we tested the hypothesis that six-hours of hypobaric hypoxia increases circulating markers of intestinal barrier injury and inflammation. A secondary aim was to determine if the changes in these markers were different between those with and without AMS. Thirteen participants were exposed to six hours of hypobaric hypoxia, simulating an altitude of 4572 m. Participants completed two 30-minute bouts of exercise during the early hours of hypoxic exposure to mimic typical activity required by those at high altitude. Pre- and post-exposure blood samples were assessed for circulating markers of intestinal barrier injury and inflammation. Data below are presented as mean ± standard deviation or median [interquartile range]. Intestinal fatty acid binding protein (Δ251 [103-410] pg•mL-1; p = 0.002, d = 0.32), lipopolysaccharide binding protein (Δ2 ± 2.4 µg•mL-1; p = 0.011; d = 0.48), tumor necrosis factor-α (Δ10.2 [3-42.2] pg•mL-1; p = 0.005; d = 0.25), interleukin-1ß (Δ1.5 [0-6.7] pg•mL-1 p = 0.042; d = 0.18), and interleukin-1 receptor agonist (Δ3.4 [0.4-5.2] pg•mL-1p = 0.002; d = 0.23) increased from pre- to post-hypoxia. Six of the 13 participants developed AMS; however, the pre- to post-hypoxia changes for each marker were not different between those with and without AMS (p > 0.05 for all indices). These data provide evidence that high altitude exposures can lead to intestinal barrier injury, which may be an important consideration for mountaineers, military personnel, wildland firefighters, and athletes who travel to high altitudes to perform physical work or exercise.

Gastrointestinal permeability and inflammatory status of preweaning dairy calves in response to decreasing the ratio of n-6 to n-3 fatty acid in milk replacer.

The ratio of n-6 to n-3 fatty acid (FA) is between 2 and 10 times higher in milk replacer (MR) than in whole milk, which may promote inflammation and compromise the integrity of the intestinal epithelium. To evaluate how decreasing the n-6:n-3 FA ratio of MR affects gastrointestinal (GIT) permeability and inflammatory status, 30 dairy calves (2.8 ± 1.06 d of age; mean ± standard deviation) were randomly assigned to be fed an MR with an n-6:n-3 FA ratio of 40:1 (CON; 29.3% crude fat of DM; n = 15) or 6.5:1 (n-3; 29.1% crude fat of DM; n = 15). Calves were fed 7.0 L/d in 2 meals. Calves were weighed and fecal consistency was analyzed weekly. On d 22, calves were administered Cr-EDTA, lactulose, and d-mannitol to assess GIT permeability. Blood and total urine were sequentially collected for 6 and 24 h, respectively, and analyzed for marker content. Whole blood collected 4 h after the meal was subjected to an ex vivo lipopolysaccharide (LPS) challenge to evaluate cytokine secretion from blood cells. Calves were euthanized on d 25 for collection of intestinal tissue samples. Tissue samples were processed to assess FA composition by gas chromatography, histomorphology by bright-field microscopy, and gene expression of tight junction proteins, lipid metabolism enzymes, and immune molecules by real-time quantitative PCR. Data were analyzed using PROC GLIMMIX in SAS (version 9.4, SAS Institute Inc.). Growth performance and fecal consistency were unaffected. Calves fed MR with a lower ratio of n-6 to n-3 FA had 2-fold higher n-3 FA contents and 2-fold lower ratios of n-6 to n-3 FA in proximal jejunum and ileum tissues. Total urinary recovery (0-24 h relative to marker administration) and plasma concentrations of the markers were unaffected. Expression of TJP1 tended to be higher in proximal jejunum tissue and lower in ileum tissue of n-3 calves. The expression of TLR4 and TNFA tended to be higher and CD14 was higher in ileum tissue of n-3 calves. Plasma concentrations of interleukin-4 were decreased in response to the ex vivo LPS challenge in n-3 calves. Histomorphology and GIT permeability were largely unaffected by treatment. Furthermore, the inclusion of linseed and algae oil may promote inflammation, as suggested by greater concentrations of the acute-phase proteins haptoglobin and serum amyloid A postprandially, demonstrating that fat sources should be evaluated for their suitability for MR formulations. Understanding how MR composition affects dairy calf health may improve nutritional strategies on farm.

Effect of forage types differing in undigested neutral detergent fiber concentration and forage inclusion rate on reticulo-ruminal motility and fermentation, total tract barrier function, and blood metabolites of finishing beef heifers.

This study evaluated the effects of forages (BarS vs. STR) that differ in the uNDF concentration and FI rate on ruminal fermentation, total tract barrier function, reticulo-ruminal motility, and blood metabolites of beef heifers. Six ruminally cannulated Hereford × Simmental heifers (699 ± 69.1 kg BW) were used in a 6 × 6 Latin square (26 d periods) with a 2 × 3 factorial treatment arrangement. However, 1 heifer was removed from the study after period 2 due to health problems unrelated to treatment, resulting in an incomplete 6 × 6 Latin square design. Barley grain-based diets were formulated using BarS or wheat STR to alter uNDF (7.1% vs. 8.5% DM) with FI rates of 5%, 10%, or 15% of DM. There were limited interactions between the forage type and FI. DM intake was not affected (P ≥ 0.10) by forage type or FI. Use of STR vs. BarS increased uNDF intake (P < 0.001). Increasing FI increased (P < 0.001) uNDF intake for those fed 15% forage. Ruminal pH was not affected (P ≥ 0.10) by forage type; however, cattle fed 5% FI had lesser (P = 0.017) mean ruminal pH and maximum pH (P = 0.018) than those fed 10% and 15% of forage. The total SCFA concentration was not affected by forage type (P = 0.84) but cattle fed the 5% FI rate had lesser (P < 0.001) molar proportion of acetate when compared with cattle fed 10% and 15% forage. Increasing the FI rate decreased the molar proportion of propionate (P < 0.001). Feeding STR relative to BarS decreased (P = 0.041) the reticulo-ruminal contraction duration. In contrast, cattle fed the 10% and 15% FI rates had a greater (P = 0.028) contraction frequency with lower (P = 0.048) contraction area than those fed 5% forage. Plasma glucose, serum insulin, and serum amyloid A were not affected by forage type or FI rate (P ≥ 0.10). Cattle fed 15% forage had lesser (P = 0.040) concentration of serum haptoglobin when compared with cattle fed 5% or 10% forage. In conclusion, forage type used to affect the dietary uNDF concentration, and FI rate act independently suggesting that the provision of STR to increase uNDF reduces reticulo-ruminal contraction duration and total tract permeability but may not affect ruminal pH. Increasing the FI increased dietary uNDF, stabilized ruminal pH, stimulated more frequent reticulo-ruminal contractions, and may decrease the permeability of the gastrointestinal tract and systemic inflammation.

As a strategy to improve performance, feedlot cattle are often fed diets containing a high proportion of grain with minimal forage. However, diets with insufficient fiber may predispose cattle to nutritional disorders, such as ruminal acidosis, which leads to poor performance and increases production costs. Given the growing concern regarding minimal fiber requirement, the present study was designed to investigate the effects of forages differing in undigested neutral detergent fiber (uNDF; barley silage [BarS] vs. straw [STR]) concentration and forage inclusion (FI) rate (5%, 10%, or 15% of dietary dry matter [DM]) on reticulo-ruminal motility and fermentation, total tract barrier function, and blood metabolites of beef cattle. The inclusion of STR increased uNDF intake and rumination rate, but decreased reticulo-ruminal contraction duration, with no effect on ruminal pH and total short-chain fatty acid (SCFA) concentration. Decreasing the FI rate decreased uNDF intake, ruminating time, ruminal pH, and reticular contraction frequency, but increased the molar proportion of propionate and concentration of serum haptoglobin. In conclusion, forage type and FI rate act independently suggesting that providing forages that increase dietary uNDF may stimulate the frequency of reticulo-ruminal contractions without affecting ruminal pH. However, increasing FI inadvertently increased dietary uNDF, stabilized ruminal pH, increased rumination time, stimulated ruminal contractions, and decreased indicators of systemic inflammation.

Clinical response in dogs with acute hemorrhagic diarrhea syndrome following randomized probiotic treatment or fecal microbiota transplant.

Probiotics and fecal microbiota transplants (FMTs) are two microbiome-targeted therapies that have been investigated for use in gastrointestinal diseases associated with dysbiosis. The aim of this study was to compare the effects of an oral multi-strain probiotic and enema-administered FMTs on clinical signs and serum lipopolysaccharide in dogs with acute hemorrhagic diarrhea syndrome (AHDS). A total of 18 client-owned dogs with a diagnosis of AHDS were enrolled in a randomized, blinded study at the time of hospital admission. The dogs were randomized into two groups: the probiotic group received a daily oral probiotic (200 × 109 CFU/10kg q 24 h) for 14 days and a single sham enema; the FMT group received a single FMT via retention enema (10 mL/kg) and placebo oral capsule for 14 days. All dogs received concurrent standard-of-care therapy, including intravenous fluids and anti-emetics; no dogs received antimicrobials. The fecal score, disease severity scores, and serum lipopolysaccharide were measured on days 0, 3, and 14. Fourteen of eighteen enrolled dogs completed the study (n = 9 probiotics; n = 5 FMT). Lipopolysaccharide decreased on days 3 and 14 from baseline and correlated with fecal and disease severity scores. There was no difference in the duration or severity of clinical signs in dogs with AHDS following an enema-administered FMT compared to probiotic treatment. Further evaluation of serum lipopolysaccharide as a marker of disease severity and recovery is warranted.

Effect of physically effective neutral detergent fiber and undigested neutral detergent fiber on eating behavior, ruminal fermentation and motility, barrier function, blood metabolites, and total tract digestibility in finishing cattle.

This study evaluated the effects of physically effective neutral detergent fiber (peNDF) and undigested neutral detergent fiber (uNDF) on eating behavior, ruminal fermentation and motility, barrier function, blood metabolites, and total tract nutrient digestibility for finishing cattle. Six Simmental heifers (668 ± 28.4 kg BW) were used in a replicated 3 × 3 Latin square (21 d periods) balanced for carry-over effects. Treatments included a control (CON; Table 1) with no forage peNDF and minimal uNDF (peNDF: 0.0%, and uNDF: 4.88 ± 0.01; 95.15% barley grain, 4.51% vitamin and mineral supplement, and 0.34% urea on a DM basis). Pelleted wheat straw (PELL) was included at 10% of dietary DM by replacing barley grain to provide added uNDF but no forage peNDF (peNDF: 0.00%, and uNDF: 6.78 ± 0.02%). Finally, chopped wheat straw (STR) was included as a replacement for pelleted wheat straw to provide forage peNDF and uNDF (peNDF: 1.74 ± 0.06%, and uNDF: 6.86 ± 0.03%). Dry matter intake was not affected (P = 0.93) by treatments. Cattle fed CON spent less time ruminating (P = 0.010) and had less meals/d (P = 0.035) when compared with cattle fed STR, with those fed PELL being intermediate but not different from other treatments. Cattle fed CON had lesser ruminal pH (P = 0.020), and a greater duration that pH was < 5.5 (P = 0.020) as compared to cattle fed STR, with those fed PELL being intermediate but not different. Cattle fed CON and PELL had greater total short-chain fatty acid concentration (P = 0.003) and molar proportion of propionate (P < 0.001) when compared with cattle fed STR. Cattle fed STR had greater (P = 0.010) total ruminal pool size when compared with cattle fed CON and PELL. Cattle fed CON had greater (P = 0.043) duration between ruminal contractions when compared with cattle fed STR, with those fed PELL being intermediate. Cattle fed CON had greater serum amyloid A (P = 0.003) and haptoglobin (P < 0.001) concentration when compared with the other treatments. Cattle fed CON had greater dry matter (P < 0.001) digestibility when compared with the other treatments. In conclusion, inclusion of PELL and STR impacted eating behavior, but only STR affected ruminal fermentation, ruminal motility, systemic inflammation, and total tract nutrient digestibility in finishing cattle. These results are interpreted to suggest that the combination of peNDF and uNDF may be better than uNDF alone to adequately capture biological effects of NDF in high-concentrate diets.

Normally forages are included at minimal levels when formulating finishing cattle diets due to lower digestibility and higher operational and economic costs than concentrates. However, insufficient fiber may increase the risk of nutritional disorders such as ruminal acidosis, negatively impacting health, and consequently growth performance of feedlot cattle. An understanding of the minimum forage requirement and the constituent factors that best explain that requirement can help to promote health and performance while minimizing cost. The present study compared the effects of physically effective neutral detergent fiber (peNDF) and undigested neutral detergent fiber (uNDF) in concentrate-based finishing diets. Feeding more peNDF increased ruminating time and ruminal pH, but decreased time between ruminal contractions, total short-chain fatty acid concentration, and the molar proportion of propionate when compared with feeding a diet with minimal peNDF and uNDF. Responses to feeding uNDF with no peNDF were generally intermediate but not different from other treatments. In addition, feeding more peNDF decreased indicators of systemic inflammation and dry matter digestibility when compared with the diet with less peNDF or uNDF. In conclusion, peNDF, or at least the combination of peNDF and uNDF may be better than uNDF alone to stimulate rumination and ruminal motility, thereby stabilizing ruminal pH. Using both peNDF and uNDF may be useful in characterizing the value of fiber in finishing diets fed to feedlot cattle.

Effects of mixed Eimeria challenge on performance, body composition, intestinal health, and expression of nutrient transporter genes of Hy-Line W-36 pullets (0-6 wks of age).

A study was aimed to investigate the effects of mixed Eimeria challenge on performance, gastrointestinal health, oxidative stress, inflammation, and expression of nutrient transporter genes of Hy-Line W-36 pullets. A total of 540, 16-d old pullets were randomly allocated into 5 treatment groups with 6 replicate cages, including a nonchallenged control group. A mixed Eimeria species solution containing 50,000 E. maxima, 50,000 E. tenella, and 250,000 E. acervulina oocysts per mL was prepared and challenged to one group as a high-dose treatment (High). The 2-fold serial dilution was done to prepare the medium-high (Med-High: 25,000 E. maxima; 25,000 E. tenella; and 125,000 E. acervulina), the medium-low (Med-Low: 12,500 E. maxima; 12,500 E. tenella; and 62,500 E. acervulina), and the low (Low: 6,250 E. maxima; 6,250 E. tenella; and 31,250 E. acervulina) dose treatments, and these dosages were challenged to 3 remaining groups, respectively. Growth performance, daily feed intake (FI), and mortality were calculated from 0-14 d postinfection (DPI). Gastrointestinal permeability (GP) was measured on 3, 5, 6, 7, and 9 DPI. The result indicated significant linear responses to the Eimeria challenge dosage in average body weight and body weight gain (P < 0.0001). An interaction between treatment and DPI was observed for FI (P < 0.0001). Feed intake significantly dropped from 4 DPI and did not recover until 12 DPI in the challenged groups. The lowest FI for each of the challenged groups was observed on 5 DPI. Gastrointestinal permeability increased linearly, peaking at 5 DPI, and was recovered back to normal by 9 DPI in the challenged groups. Furthermore, gene expression of tight junction proteins was linearly upregulated by increased Eimeria dosages. The oxidative status of the pullets was lowered in the challenged groups than the nonchallenged control group, whereas the expression of inflammatory and proinflammatory cytokines was upregulated by Eimeria challenge on 6 DPI (P < 0.05). The highest mortality was observed in pullets challenged with the High, followed by the Med-High (P < 0.0001) on 5 DPI. In summary, the mixed Eimeria challenge linearly reduced the growth performance of pullets with an increase in oxidative stress and inflammation. A severe effect of Eimeria on gastrointestinal health was observed on 5 or 6 DPI as suggested by GP, tight junction genes, and mortality results. This study indicates that Eimeria infection can be a threat to gastrointestinal health related issues in pullets.

The effect of prolonged interval and continuous exercise in the heat on circulatory markers of intestinal barrier integrity.

PURPOSE: The purpose of this study was to determine the effect of prolonged high-intensity interval (INT) and moderate-intensity continuous (CONT) treadmill exercise in the heat on markers of enterocyte injury and bacterial endotoxin translocation. METHODS: Nine males completed 2 h of work-matched exercise in the heat (40 °C and 15% RH) as either INT (2 min at 80% VO2max and 3 min at 30% VO2max) or CONT (~ 50% of VO2max). Blood samples collected pre- and post-exercise were assayed for intestinal fatty acid-binding protein (I-FABP), claudin-3 (CLDN-3), and lipopolysaccharide-binding protein (LBP). RESULTS: I-FABP was significantly increased from pre- to post-exercise in CONT (913.96 ± 625.13 to 1477.26 ± 760.99 pg•mL-1; p = 0.014, d = 0.766) and INT (714.59 ± 470.27 to 1547.93 ± 760.99 pg•mL-1; p = 0.001, d = 1.160). Pre- to post-exercise changes in I-FABP were not different between CONT and INT (p = 0.088, d = 0.414). LBP was significantly increased from pre- to post-exercise in INT (15.94 ± 2.90 to 17.35 ± 3.26 µg•mL-1; p = 0.028, d = 0.459) but not CONT (18.11 ± 5.35 to 16.93 ± 5.39 µg•mL-1; p = 0.070, d = 0.226), and pre- to post-exercise changes in LBP were higher in the INT compared to CONT (p < 0.001, d = 1.160). No significant changes were detected from pre- to post-exercise for CLDN-3 in CONT (14.90 ± 2.21 to 15.30 ± 3.07 µg•mL-1) or INT (15.55 ± 1.63 to 16.41 ± 2.11 µg•mL-1) (p > 0.05). CONCLUSIONS: We conclude that prolonged exercise in the heat induces enterocyte injury, but interval (or intermittent) exercise may cause greater bacterial endotoxin translocation which may increase the risk for local and systemic inflammation.

Improving Aqueous Solubility and In Vitro Pharmacokinetic Properties of the 3-Nitroimidazo[1,2-a]pyridine Antileishmanial Pharmacophore.

An antileishmanial structure−activity relationship (SAR) study focused on positions 2 and 8 of the imidazo[1,2-a]pyridine ring was conducted through the synthesis of 22 new derivatives. After being screened on the promatigote and axenic amastigote stages of Leishmania donovani and L. infantum, the best compounds were tested against the intracellular amastigote stage of L. infantum and evaluated regarding their in vitro physicochemical and pharmacokinetic properties, leading to the discovery of a new antileishmanial6-chloro-3-nitro-8-(pyridin-4-yl)-2-[(3,3,3-trifluoropropylsulfonyl)methyl]imidazo[1,2-a]pyridine hit. It displayed low cytotoxicities on both HepG2 and THP1 cell lines (CC50 > 100 µM) associated with a good activity against the intracellular amastigote stage of L. infantum (EC50 = 3.7 µM versus 0.4 and 15.9 µM for miltefosine and fexinidazole, used as antileishmanial drug references). Moreover, in comparison with previously reported derivatives in the studied series, this compound displayed greatly improved aqueous solubility, good mouse microsomal stability (T1/2 > 40 min) and high gastrointestinal permeability in a PAMPA model, making it an ideal candidate for further in vivo studies on an infectious mouse model.

Safety of Bifidobacterium breve, Bif195, employing a human exercise-induced intestinal permeability model: a randomised, double-blinded, placebo-controlled, parallel group trial.

We have previously shown that the probiotic Bifidobacterium breve strain Bif195 alleviates mucosal injury including ulcer formation in the upper intestine induced by non-steroid anti-inflammatory drugs (NSAIDs). Here, we report additional safety use of Bif195 in 126 healthy humans undergoing an exercise-induced intestinal permeability challenge in a double-blinded, placebo-controlled randomised 6-week intervention trial. Intestinal permeability was assessed by urinary lactulose/rhamnose (L/R) ratio. L/R ratio, plasma intestinal fatty acid binding protein (I-FABP) and gastrointestinal symptom rating scale (GSRS) questionnaire were measured resting and after a 1 h treadmill challenge, prior to and at the end of the intervention. To be able to compare the equivalence of resting state at baseline, of this cohort of well-trained subjects, to non-trained subjects, a cohort of 63 healthy and non-trained subjects (<2 h/week of endurance sports) was included. Study subjects (well-trained) were 35.7% women with a mean age and body mass index (in kg/m2) of 35.0 years and 24.8, respectively. There were no differences between the Bif195 and placebo groups in effects on L/R ratio, I-FABP and GSRS questionnaire score. In addition, there were no differences between Bif195 and placebo in number of adverse events and change in cytokines, liver or kidney biomarkers. The exercise model successfully induced intestinal permeability by statistically significantly increasing L/R ratio by ~100% (P<0.0001) and cytokines after the exercise challenge. No significant difference was found between well-trained and non-trained subjects in baseline resting L/R ratio. In conclusion, the reported cytoprotective effects of Bif195 are unlikely to be primarily related to small bowel permeability, and the safety of Bif195 in individuals with increased permeability is supported by the present data. ClinicalTrials.gov: NCT03027583.

Technical Note: A procedure to place urinary catheters in 1- and 6-week-old preweaned Holstein heifer calves for the in vivo evaluation of intestinal permeability.

Oral administration of indigestible markers and subsequent urine collection is a useful method to determine in vivo gastrointestinal tract (GIT) permeability in cattle for research purposes. However, urine sampling techniques often rely on total waste collection, which reduces the ability to perform more frequent sampling and obtain accurate volumes and sterile samples. An alternative is urethral catheterization, though the feasibility of this technique has not been thoroughly tested in preweaned Holstein heifer calves. The study objective was to develop a urethral catheter placement procedure in preweaned Holstein heifer calves for continuous and accurate urine collection to evaluate GIT permeability using an indigestible marker. Fifteen Holstein heifer calves had catheters placed at approximately 1 wk (8.0 ± 1.5 d) and 6 wk (40.0 ± 1.5 d) of age. During the procedure, calves were individually housed and restrained. The vulva was sterilized and then a sterile, lubricated speculum was inserted into the vagina. A sterile 0.09 cm diameter guidewire was guided into a lubricated, sterile 10 French Foley catheter. The catheter was inserted at approximately 5 through 7 cm into the urethral opening, guided into the bladder, and the catheter balloon was filled with 10 mL of water. The guidewire was removed, and urine flow confirmed correct placement before a 4-L urinary drainage bag was attached to the catheter. After catheterization (24 h), 1 L of chromium (Cr)-ethylenediaminetetraacetic acid was orally dosed to the calves. Calf health observations were made six times over a 48-h period, and any occurrence of vaginal discharge, tissue discharge in catheter, bleeding, inflammation, or abnormal urine was considered a localized reaction. The proportion of localized reactions for each age group was determined using Microsoft Excel, and the total Cr output was analyzed using PROC GLIMMIX. Localized reactions occurred for 20.0% of the 1-wk-old calves and 13.3% of the 6-wk-old calves. In the first 4 h, urine was collected every 15 min, and there were no overall Cr output differences (P = 0.38; 10.28 ± 3.21 mg Cr) when comparing 1- and 6-wk-old calves. However, 1-wk-old calves tended (P = 0.08) to have greater overall Cr output at 480 min (19.2%) and 1,440 min (41.9%) when compared with 6-wk-old calves. In summary, urinary catheterization is a viable urinary collection method for the determination of in vivo GIT permeability in preweaned Holstein heifer calves.

Neonatal calves are highly susceptible to enteric disease during their first few weeks of life, and enteric disease is the leading cause of preweaning morbidity and mortality. A consequence of enteric disease is greater gastrointestinal tract (GIT) permeability in neonatal calves, which is also influenced by reduced intestinal maturity and environmental factors. Therefore, an accurate and precise method of evaluating GIT permeability in neonatal calves is necessary to develop appropriate treatment and mitigation strategies. The oral administration of indigestible markers and measurement of their presence in urine is an accepted method to determine the total GIT permeability in mature heifers. However, current urine collection methods in preweaned heifer calves may not be reliable. Therefore, the study objective was to develop a urinary catheterization method to collect urine accurately and precisely for the in vivo determination of GIT permeability in 1- and 6-wk-old Holstein heifer calves. It was determined that the urinary catheterization procedure and collection system developed in this study were viable and could be applied when evaluating GIT permeability in preweaned Holstein heifer calves using orally dosed indigestible markers.

Role of the gut microbiome in three major psychiatric disorders.

Major depressive disorder (MDD), bipolar disorder (BD) and schizophrenia-spectrum disorders (SSD) are heterogeneous psychiatric disorders, which place significant burden on patient's well-being and global health. Disruptions in the gut-microbiome may play a role in these psychiatric disorders. This review presents current data on composition of the human gastrointestinal microbiota, and its interaction mechanisms in the gut-brain axis in MDD, BD and SSD. Diversity metrics and microbial relative abundance differed across studies. More studies reported inconsistent findings (n = 7) or no differences (n = 8) than studies who reported lower α-diversity in these psychiatric disorders (n = 5). The most consistent findings across studies were higher relative abundances of the genera Streptococcus, Lactobacillus, and Eggerthella and lower relative abundance of the butyrate producing Faecalibacterium in patients with psychiatric disorders. All three increased genera were associated with higher symptom severity. Confounders, such as medication use and life style have not been accounted for. So far, the results of probiotics trials have been inconsistent. Most traditional and widely used probiotics (consisting of Bifidobacterium spp. and Lactobacillus spp.) are safe, however, they do not correct potential microbiota disbalances in these disorders. Findings on prebiotics and faecal microbiota transplantation (FMT) are too limited to draw definitive conclusions. Disease-specific pro/prebiotic treatment or even FMT could be auspicious interventions for prevention and therapy for psychiatric disorders and should be investigated in future trials.

High-altitude exposures and intestinal barrier dysfunction.

Gastrointestinal complaints are often reported during ascents to high altitude (>2,500 m), though their etiology is not known. One potential explanation is injury to the intestinal barrier which has been implicated in the pathophysiology of several diseases. High-altitude exposures can reduce splanchnic perfusion and blood oxygen levels causing hypoxic and oxidative stress. These stressors might injure the intestinal barrier leading to consequences such as bacterial translocation and local/systemic inflammatory responses. The purpose of this mini-review is to 1) discuss the impact of high-altitude exposures on intestinal barrier dysfunction and 2) present medications and dietary supplements which may have relevant impacts on the intestinal barrier during high-altitude exposures. There is a small but growing body of evidence which shows that acute exposures to high altitudes can damage the intestinal barrier. Initial data also suggest that prolonged hypoxic exposures can compromise the intestinal barrier through alterations in immunological function, microbiota, or mucosal layers. Exertion may worsen high-altitude-related intestinal injury via additional reductions in splanchnic circulation and greater hypoxemia. Collectively these responses can result in increased intestinal permeability and bacterial translocation causing local and systemic inflammation. More research is needed to determine the impact of various medications and dietary supplements on the intestinal barrier during high-altitude exposures.

Simulation of feed restriction and fasting: Effects on animal recovery and gastrointestinal permeability in unweaned Angus-Holstein calves.

Feed restriction and fasting experienced during commercial production negatively affect unweaned calves' behavior and health status. Transportation and stays at assembly centers are the main factors generating these disorders. For this study, 20 unweaned Angus-Holstein bull calves [44.1 ± 2.04 kg of body weight (BW) and 14.7 ± 0.63 d of age (± standard error)] were used to evaluate the effects of feed restriction and fasting on performance, energy status [serum concentration of glucose, ß-hydroxybutyrate (BHB), and nonesterified fatty acids], and gastrointestinal permeability [serum concentration of citrulline, chromium (Cr)-EDTA, lactulose, and d-mannitol]. Calves were randomly assigned to 1 of 4 treatments that simulated the feed restrictions of an assembly center situation on one hand, and the fasting hours during transportation on the other. Treatments were as follows. Control (CT): from d -4 to -1, calves were fed 2.5 L of milk replacer (MR) twice daily; concentrate and straw were offered ad libitum. Mild (MD): calves were fed only MR (d -4 to -1) as described for CT, and on d -1 calves were subjected to a 9-h feed withdrawal. Moderate (MO): calves were fed only MR (d -4 to -1) as described for CT and on d -1 subjected to a 19-h feed withdrawal. Severe (SV): calves were fed only 2.5 L of a rehydrating solution twice daily (d -4 to -1) and on d -1 subjected to a 19-h feed withdrawal. From d 0 to d 42 (weaning) all calves were fed the same feeding program (MR, concentrate, and straw ad libitum). Results showed that BW was greater for the CT treatment compared with the others from d 0 to d 7, whereas BW of SV was lesser compared with the others from d -1 to d 7. No differences among treatments were observed at weaning. At d 2 concentrate intakes of MD, MO, and SV were lesser compared with CT. By d 4, concentrate intake of SV was similar to that for CT and greater than MD and MO. Similarly to BW, no differences in concentrate intake among treatments were observed at weaning on d 42 of the study. At d -1 for SV and d 0 in all restricted calves, serum glucose concentration was lesser compared with CT. At d -1 and 0, nonesterified fatty acids and BHB serum concentrations were greater in the SV calves compared with the other treatments. By d 2, serum concentrations of nonesterified fatty acids, BHB, and glucose were restored to CT levels. At d -1 serum citrulline concentration was lesser in SV and greater in MD calves. The CT calves had lower serum concentrations of Cr-EDTA (d -1 and d 0), lactulose (d 0), and d-mannitol (d 0) compared with the other restricted calves. Results showed that degree of dietary restriction, type of liquid diet (MR or rehydrating solution), and fasting hours (9 vs. 19 h) affected calves' BW, concentrate intake, and serum concentration of markers indicative of energy status and gastrointestinal permeability.

Intestinal adaptations to energy source of milk replacer in neonatal dairy calves.

Most milk replacers (MR) contain more lactose compared with whole milk, which, when fed at a large meal size, could influence gut barrier function in calves. This study evaluated how replacing lactose in MR with fat (on a wt/wt basis) affects intestinal histomorphology and permeability in neonatal dairy calves. Thirty-four Holstein-Friesian bull calves were blocked by dam parity and randomly assigned to 1 of 2 treatments (n = 17): a high-lactose (46.1% lactose, 18.0% crude fat, and 23.9% crude protein of dry matter) or a high-fat MR (HF; 39.9% lactose, 24.6% crude fat, and 24.0% crude protein of dry matter). Calves were individually housed and fed pooled colostrum at 1.5 h and 12 h postnatally, at 18 and 9% of metabolic body weight (BW0.75), respectively. From 24 h postnatally until the end of the study (d 7), calves were transitioned to be fed MR (prepared at 15% solids) at 18% of BW0.75 twice daily at 0700 and 1900 h. During postprandial sampling on d 6, intestinal permeability was assessed by mixing lactulose (1.03 g/kg of BW0.75) and d-mannitol (0.31 g/kg of BW0.75) into the morning meal without altering total meal volume. Sequential blood samples were collected via jugular catheter, and total urine was collected for 12 h to measure the marker content. Calves were euthanized 3 h after the morning meal on d 7, and gastrointestinal tract tissues and digesta were collected for analysis of histomorphology, digesta osmolality, and gene expression. The empty gastrointestinal tracts of HF calves were heavier, although length did not differ and differences in histomorphology were minor. Digesta osmolality changed along the tract without differences between treatments. Plasma lactulose was greater in HF, although plasma d-mannitol and the recovery of both markers in urine were unaffected. No significant differences were detected in gene expression, although HF calves tended to have lower expression of TJP1 and CLDN2 and higher expression of proinflammatory cytokine IL1B in ileum tissue. In conclusion, partially replacing lactose in MR with fat resulted in a heavier and more permeable gut, with minor histomorphological differences.

Impacts of increasing challenge with Eimeria maxima on the growth performance and gene expression of biomarkers associated with intestinal integrity and nutrient transporters.

This study was conducted to investigate the impacts of graded severity of Eimeria maxima infection on the growth performance and intestine health of broiler chickens. Four different levels of E. maxima-challenged treatments were used, including a non-challenged control group, a low challenge (12 500 oocysts), a medium challenge (25 000 oocysts), and a high challenge dose (50 000 oocysts). There were eight replicate cages per treatment, with 12 birds in each cage, and chickens in the challenged groups orally received sporulated oocysts on day 14. Gastrointestinal permeability was measured by fluorescein isothiocyanate dextran at 5 days post-infection (dpi), whereas intestinal morphology and gene expression of nutrient transporters and tight junction proteins were determined at 6 dpi. The results demonstrate a linear reduction in growth performance, jejunal villus height, and jejunal integrity with graded challenge doses of E. maxima (P < 0.01). Moreover, linear regulation of nutrient transporters and tight junction proteins was a consequence of increasing Eimeria infection levels (P < 0.01). The linear increase of Claudin 1, cationic amino acid transporter, glucose transporter 1, and L-type amino acid transporter genes was associated with increased severity of coccidiosis (P < 0.01). Furthermore, expression of nutrient transporters located at the brush border membrane were down-regulated (P < 0.01) with increasing E. maxima inoculation dose. In conclusion, growth performance and key intestinal integrity biomarkers in broiler chickens were adversely influenced in a dose-dependent manner by E. maxima infection.

Whole and Isolated Protein Fractions Differentially Affect Gastrointestinal Integrity Markers in C57Bl/6 Mice Fed Diets with a Moderate-Fat Content.

Various proteins or protein fractions reportedly positively affect gastrointestinal integrity and inflammation in diets providing >45% energy as fat. This study tested whether benefits were seen in diets providing 30% of energy as fat. Purified diets (PD) with isolated soy protein (ISP), dried whole milk powder (DWMP), milk fat globule membrane (MFGM), or milk protein concentrate (MPC) as protein sources were fed to C57BL/6J mice (n = 15/diet group) for 13 weeks. MFGM-fed mice were heaviest (p < 0.005) but remained within breeder norms. Growth rates and gut motility were similar for all PD-fed mice. FITC-dextran assessed gut permeability was lowest in DWMP and MFGM (p = 0.054); overall, plasma endotoxin and unprovoked circulating cytokines indicated a non-inflammatory state for all PD-fed mice. Despite differences in cecal butyrate and intestinal gene expression, all PDs supported gastrointestinal health. Whole milk provided more positive effects compared to its fractions. However, ISP-fed mice showed a >370%, (p < 0.006) increase in colonic myeloperoxidase activity indicative of tissue neutrophil infiltration. Surprisingly, FITC-dextran and endotoxin outcomes were many folds better in PD-fed mice than mice (strain, vendor, age and sex matched) fed a "chow-type" nutritionally adequate non-PD. Additional variables within a diet's matrix appear to affect routine indicators or gastrointestinal health.

Activation of the Innate Immune System in Children With Irritable Bowel Syndrome Evidenced by Increased Fecal Human ß-Defensin-2.

BACKGROUND & AIMS: The role of the innate immune system in functional gastrointestinal pain disorders is unclear. We investigated the role of ß-defensin-2 and gut permeability in childhood irritable bowel syndrome (IBS) and functional abdominal pain (FAP) symptom generation. METHODS: Fecal ß-defensin-2 (and in a subset, gut permeability) was measured in children with IBS (n = 116), FAP (n = 33), and healthy control (HC) children (n = 72). IBS and FAP patients were recruited from tertiary and primary care, and HCs were recruited from primary care. RESULTS: ß-defensin-2 concentration was greater in children with IBS (P = .003) and FAP (P = .03) than in HCs. ß-defensin-2 was greater in girls with IBS than female HCs (P = .007) and in girls with IBS vs boys with IBS (P = .036). There was no difference by sex in the FAP and HC groups. For the entire cohort, ß-defensin-2 correlated with multiple pain symptoms. In the IBS group, ß-defensin-2 correlated with pain interference (P = .014). No correlation with pain was found in the FAP or HC group. Gut permeability was greater in the IBS vs the FAP and HC groups (P = .038). For the entire cohort, permeability correlated with the number of pain episodes (P = .041) and interfering pain episodes (P = .049). For the entire cohort there was a correlation between ß-defensin-2 and permeability (P = .003), with borderline correlation in the IBS group (P = .086). For the cohort and IBS and HC groups, the number of bowel movements was modestly inversely related to fecal ß-defensin-2 concentrations. CONCLUSIONS: Increased fecal ß-defensin-2 concentration in children with IBS suggests activation of the innate immune system in some, which, along with increased gut permeability, appears related to abdominal pain symptoms. Sex is an important variable in interpreting ß-defensin-2 concentration in children with IBS.

Graded Eimeria challenge linearly regulated growth performance, dynamic change of gastrointestinal permeability, apparent ileal digestibility, intestinal morphology, and tight junctions of broiler chickens.

This study was conducted to evaluate graded Eimeria challenge on growth performance, apparent ileal digestibility, gastrointestinal permeability, intestinal morphology, gene expression of tight junction protein, and intestinal lesion scores in broiler chickens. There were 5 groups in this study, including a control and 4 different Eimeria treatment doses. A mixed Eimeria spp. solution with 50,000 Eimeria maxima, 50,000 Eimeria tenella, and 250,000 Eimeria acervulina per milliliter was prepared for the high-dose challenge treatment. The 2-fold serial dilution was used to make the medium-high (25,000 E. maxima; 25,000 E. tenella; 125,000 E. acervulina), the medium-low (12,500 E. maxima; 12,500 E. tenella; 62,500 E. acervulina), and the low challenge dose (6,250 E. maxima; 6,250 E. tenella; 31,250 E. acervulina). A total of three hundred sixty 13-day-old male broiler chickens were randomly allocated into 5 treatments with 6 replicated cages. Growth performance was calculated from 0 to 6 D postinfection (DPI). Intestine lesion was scored on 6 DPI. Gastrointestinal permeability was measured on 3, 5, 6, 7, and 9 DPI. The results indicated significant linear reduction in growth performance, intestinal villi height, and ileal nutrient digestibility in response to the increase of Eimeria challenge dose. Furthermore, gene expression of tight junction protein was linearly upregulated by the increasing challenge doses. Significant linear increases of gastrointestinal permeability were found on 5, 6, and 7 DPI (P < 0.01). On 9 DPI, the gastrointestinal permeability was recovered back to normal level in the challenge groups. In conclusion, the higher Eimeria doses birds received, the more severe intestine damage was observed in several gastrointestinal health parameters. The medium-low or medium-high levels of mixed Eimeria oocysts is suggested as an optimum Eimeria-challenge dose to establish a subclinical challenge model for future studies evaluating nutritional strategies. Moreover, it is recommended to measure gastrointestinal permeability on 5 DPI with higher oocysts doses and 6 DPI when using the lower oocysts doses.

UCC118 supplementation reduces exercise-induced gastrointestinal permeability and remodels the gut microbiome in healthy humans.

Dysregulation of gut microbiota and intestinal barrier function has emerged as potential mechanisms underlying digestive diseases, yet targeted therapies are lacking The purpose of this investigation was to assess the efficacy of UCC118, a characterized probiotic strain, on exercise-induced GI permeability in healthy humans. In a randomized, double-blind, placebo-controlled crossover study, seven healthy adults received 4 weeks of daily UCC118 or placebo supplementation. GI hyperpermeability was induced by strenuous treadmill running performed before and after each supplementation period. While running, participants ingested 5 g of lactulose, rhamnose, and sucrose. Urine was collected before, immediately after, and every hour for 5 h after exercise to assess GI permeability. Metagenomic sequencing was performed on fecal homogenates collected prior to exercise to identify changes in microbial diversity and taxon abundances. Inflammatory biomarkers were assessed from blood and fecal homogenates collected prior to and immediately following the cessation of exercise. Exercise significantly induced intestinal permeability of lactulose, rhamnose, and sucrose (P < 0.001). UCC118 significantly reduced sucrose (Δ = -0.38 ± 0.13 vs. 1.69 ± 0.79; P < 0.05) recovery, with no substantial change in lactulose (Δ = -0.07 ± 0.23 vs. 0.35 ± 0.15; P = 0.16) or rhamnose (Δ = -0.06 ± 0.22 vs. 0.48 ± 0.28; P = 0.22). Taxonomic sequencing revealed 99 differentially regulated bacteria spanning 6 taxonomic ranks (P < 0.05) after UCC118 supplementation. No differences in plasma IL-6 or fecal zonulin were observed after UCC118 supplementation. The results described herein provide proof of principle that 4 weeks of UCC118 supplementation attenuated exercise-induced intestinal hyperpermeability. Further research is warranted to investigate the as-yet-to-be defined molecular processes of intestinal hyperpermeability and the effects of probiotic supplementation.