Short-chain fatty acids promote jejunal barrier function and caecal muscle contractibility in laying hens ex vivo.

1. Short-chain fatty acids (SCFA) exert beneficial actions in the gut; nevertheless, information about the effect of SCFA on physiological responses in the small intestine of chickens is rare.2. The aim of this study was to assess the effect of 1) different molar acetate:butyrate ratios (Ac:But; Experiment 1; 78.5% acetate and 7.3% butyrate versus 71.4% acetate and 14.0% butyrate) and 2) SCFA concentrations (Experiment 2; final concentration in chambers: 70.5 versus 141 µmol SCFA/ml buffer) on the jejunal and caecal contractibility and jejunal barrier function in laying hens. The change in muscle contractibility due to the SCFA was measured in mid-jejunal and caecal segments (n = 4 each per hen) from four laying hens using the organ bath system after precontraction with acetylcholine for 15 min. Changes in short-circuit current (ISC) and transepithelial tissue conductivity (GT) as indicators for net ion flux and barrier function, respectively, were measured in mid-jejunal tissue (n = 3/hen and treatment), mounted into Ussing chambers.3. In Experiment 1, the addition of SCFA, irrespective of the Ac:But ratio, decreased jejunal muscle tension (P < 0.05), jejunal GT as well as caused a less negative ISC (P < 0.05). In Experiment 2, the increasing SCFA concentrations increased the caecal muscle contraction and jejunal ISC by 75.6% while decreasing the GT by up to 19.6% (P < 0.05).4. In conclusion, results demonstrate that increasing butyrate proportions and SCFA concentrations stimulate caecal muscle contraction, thereby increasing caecal mixing and emptying in vivo. Jejunal ISC and GT support a strong SCFA sensing capacity in the jejunum, as both, more butyrate and higher SCFA, increased mucosal ion uptake and barrier function.

Cinnamon bark oil and coconut oil emulsions modified small intestinal motility and barrier function in laying hens in an ex vivo experiment.

1. Plant extracts and oils are supplemented in diets for chickens due to their antimicrobial capacities; however, little information exists whether they influence intestinal motility and barrier function.2. The present study aimed to determine the effect of increasing levels of cinnamon bark oil (CBO; 0%, 0.038%, 0.076% and 0.151%) and coconut oil emulsions prepared with soy and sunflower lecithin on the contractile function of enteric wall muscles in the jejunum and ileum and jejunal barrier function in laying hens.3. For testing muscle contraction, mid-jejunal and ileal segments (n = 4 each per hen) from four laying hens were placed in a longitudinal orientation into isolated organ baths filled with Krebs buffer and fastened to force transducers. Muscle segments were induced to contract with acetylcholine and the effects of the oil emulsions on contraction were measured.4. For barrier function, distal jejunal pieces were stripped of serosa before mounting into Ussing chambers and recording changes in short-circuit current (ISC) and transepithelial tissue conductivity (GT) before and after addition of the respective emulsion.5. The CBO decreased the muscle tone, representing a relaxation of on average 36.2% and 42.6% for the jejunum and ileum, respectively, compared to before the addition (P < 0.001). Moreover, CBO linearly decreased the ISC and GT of the jejunal mucosa, indicating a greater absorption of anions and increased barrier function (P < 0.001). Only the coconut oil-sunflower lecithin emulsion relaxed the muscles, whereas both coconut oil-lecithin emulsions increased the ISC but reduced the GT of the jejunal mucosa, which suggested an increased cation absorption and decreased paracellular permeability, respectively (P < 0.05).6. In conclusion, CBO and coconut oil-lecithin emulsions showed the potential to increase jejunal barrier function, whereas CBO may be more efficacious to slow down digesta passage in the small intestine.

Resistant starch reduces large intestinal pH and promotes fecal lactobacilli and bifidobacteria in pigs.

Dietary resistant starch (RS) may have prebiotic properties but its effects on fermentation and the microbial population are inconsistent. This meta-analysis aimed to quantify the relationship between RS type 2 (RS2) and intestinal short-chain fatty acids (SCFA) and pH as well as certain key bacterial taxa for intestinal health in pigs. From the 24 included articles with sufficient information about the animal, and dietary and physiological measurements published between 2000 and 2017, individual sub-data sets for fermentation metabolites, pH, bacterial abundances and apparent total tract digestibility were built and used to parameterize prediction models on the effect of RS2, accounting for inter- and intra-study variability. In addition, the effect of pig's BW at the start of the experiment and duration of the experimental period on response variables were also evaluated using backward elimination analysis. Dietary RS levels ranged from 0% to 78.0% RS, with median and mean RS levels of 28.8% and 23.0%, respectively. Negative relationships could be established between dietary RS and pH in the large intestine (P<0.05), with a stronger effect in the mid and distal colon, and feces (R 2=0.64 to 0.81; P<0.001). A dietary level of 15% RS would lower the pH in the proximal, mid-, distal colon and feces by 0.2, 0.6, 0.4 and 0.6 units, respectively. Increasing RS levels, however, did not affect SCFA concentrations in the hindgut, but enhanced the molar proportion of propionate in mid-colon and reduced those of acetate in mid-colon and of butyrate in mid- and distal colon (R 2=0.46 to 0.52; P<0.05). Backward elimination indicated an age-related decrease in mid-colonic propionate proportion and increase in mid- and distal colonic butyrate proportion (P<0.05), thereby modulating RS2 effects. In feces, increasing RS levels promoted fecal lactobacilli (R 2=0.46; P<0.01) and bifidobacteria (R 2=0.57; P<0.01), whereby the slope showed the need for a minimal RS level of 10% for a 0.5 log unit-increase in their abundance. Best-fit equations further supported that a longer experimental period increased fecal lactobacilli but decreased fecal bifidobacteria (P<0.05). In conclusion, dietary RS2 seems to effectively decrease digesta pH throughout the large intestine and increase lactic acid-producing bacteria in feces of pigs which may limit the growth of opportunistic pathogens in the hindgut. To achieve these physiologically relevant changes, dietary RS should surpass 10% to 15%.

Changes in the Rumen Epithelial Microbiota of Cattle and Host Gene Expression in Response to Alterations in Dietary Carbohydrate Composition.

The inclusion of high-quality hay (HQH), in place of concentrates, shifts dietary carbohydrate intake, and the extent to which these shifts effect epimural microbiota and epithelial gene expression of the rumen has not yet been evaluated. Eight ruminally cannulated nonlactating Holstein cows were used in a replicated 4 by 4 Latin square design with four dietary treatments containing HQH, with either 0% concentrate/100% HQH (100HQH), 25% concentrate/75% HQH (75HQH), or 40% concentrate/60% HQH (60HQH). The fourth group (control [CON]) was fed 60% normal fiber-rich hay and 40% concentrate. The data showed that measures of diversity for the rumen epimural population, specifically the Shannon (P = 0.004) and Simpson (P = 0.003) indices, decreased with increasing levels of HQH in the diet. The feeding of HQH shifted the epimural population from predominantly Firmicutes to Proteobacteria Phylogenetic analysis revealed that HQH feeding markedly shifted the abundance of Campylobacter spp. from 7.8 up to 33.5% (P < 0.001), with greater ingestion of protein (r = 0.63) and sugars (r = 0.65) in HQH diet being responsible for this shift. The expression of genes targeting intracellular pH regulation, barrier function, and nutrient uptake of rumen epithelium remained stable regardless of the carbohydrate source. In conclusion, the data suggest strong alterations of the ruminal epimural microbiota in response to changes in the nutritive patterns of the diet. Further research is warranted to evaluate the long-term effects of these significant microbial changes on rumen health and food safety aspects in cattle at a transcriptional level.IMPORTANCE Feeding of forages versus starchy concentrates is a highly debated topic. Hay is believed to be healthier and more ecological sustainable for cattle than are concentrates, although the effects of feeding hay with enhanced sugar and protein content on epimural microbiota and host gene expression have not yet been evaluated. This research provides a report of the role of feeding hay with increased sugar and protein content in place of starchy concentrates in altering epimural microbiota and in generating a host response. Our research shows that the addition of high-quality hay to dairy rations shifted nutrient intake, resulting in strong alterations in the epimural microbiota in cattle. This work provides a background for further long-term research regarding the effects of feeding practices on the host-microbiome interaction and its role in rumen health and food safety in cattle.

Differences in intestinal size, structure, and function contributing to feed efficiency in broiler chickens reared at geographically distant locations.

The contribution of the intestinal tract to differences in residual feed intake (RFI) has been inconclusively studied in chickens so far. It is also not clear if RFI-related differences in intestinal function are similar in chickens raised in different environments. The objective was to investigate differences in nutrient retention, visceral organ size, intestinal morphology, jejunal permeability and expression of genes related to barrier function, and innate immune response in chickens of diverging RFI raised at 2 locations (L1: Austria; L2: UK). The experimental protocol was similar, and the same dietary formulation was fed at the 2 locations. Individual BW and feed intake (FI) of chickens (Cobb 500FF) were recorded from d 7 of life. At 5 wk of life, chickens (L1, n = 157; L2 = 192) were ranked according to their RFI, and low, medium, and high RFI chickens were selected (n = 9/RFI group, sex, and location). RFI values were similar between locations within the same RFI group and increased by 446 and 464 g from low to high RFI in females and males, respectively. Location, but not RFI rank, affected growth, nutrient retention, size of the intestine, and jejunal disaccharidase activity. Chickens from L2 had lower total body weight gain and mucosal enzyme activity but higher nutrient retention and longer intestines than chickens at L1. Parameters determined only at L1 showed increased crypt depth in the duodenum and jejunum and enhanced paracellular permeability in low vs. high RFI females. Jejunal expression of IL1B was lower in low vs. high RFI females at L2, whereas that of TLR4 at L1 and MCT1 at both locations was higher in low vs. high RFI males. Correlation analysis between intestinal parameters and feed efficiency metrics indicated that feed conversion ratio was more correlated to intestinal size and function than was RFI. In conclusion, the rearing environment greatly affected intestinal size and function, thereby contributing to the variation in chicken RFI observed across locations.

Assessing serum metabolite profiles as predictors for feed efficiency in broiler chickens reared at geographically distant locations.

1. The objective of this study was to investigate differences in growth performance, serum intermediary metabolites, acute-phase proteins and white blood cells in low, medium and high-residual feed intake (RFI) chickens. It was also assessed if the environment affects the feed efficiency (FE) and FE-related performance and serum profiles of chickens. 2. Individual body weight (BW) and feed intake (FI) were recorded from d 7 of life. At 5 weeks of age, female and male broiler chickens (Cobb 500) were selected according to their RFI (L1: Austria; L2: UK; n = 9/RFI group, sex and locatity -45on) and blood samples were collected. 3. Chickens at L1 had similar FI but a 15% higher BW gain compared to chickens at L2. The RFI values of female chickens were -231, 8 and 215 g and those of male chickens -197, 0 and 267 g for low, medium and high RFI, respectively. 4. Location affected serum glucose, urea, cholesterol, non-esterified fatty acids (NEFA) and ovotransferrin in females, and serum glucose and triglycerides in male chickens. Serum uric acid and NEFA linearly increased from low to high RFI in females, whereas in males, cholesterol showed the same linear response from low to high RFI. Serum alpha-1-acid glycoprotein and blood heterophil-to-lymphocyte ratio linearly increased by 35% and 68%, respectively, from low to high RFI but only in male chickens at L1. 5. Regression analysis showed significant positive relationships between RFI and serum uric acid (R2 = 0.49) and cholesterol (R2 = 0.13). 6. It was concluded that RFI-related variation in serum metabolites of chickens was largely similar for the two environments and that serum metabolite patterns could be used to predict RFI in chickens.

Feed efficiency metrics in growing pigs.

The objective of the present study was to quantify the interrelationships between different feed efficiency measures in growing pigs and characterize pigs divergent for a selection of these measures. The data set included data from 311 growing pigs between 42 and 91 d of age from 3 separate batches. Growth-related metrics available included midtest metabolic BW (BW), energy intake (EI), and ADG. Ratio efficiency traits included energy conversion ratio (ECR), Kleiber ratio (ADG/BW), relative growth rate (RGR), residual EI (REI), and residual daily gain (RDG). Residual intake and gain (RIG; i.e., a dual index of both REI and RDG) and residual midtest metabolic weight (RMW) were also calculated. Simple Pearson correlations were estimated between the growth and feed efficiency metrics. In litters with at least 3 pigs of each sex, pigs were separately stratified on each residual trait as high, medium, and low rank. Considerable interanimal variability existed in all metrics evaluated. Male pigs were superior to females for all metrics ( < 0.001) except for both BW and EI, where no sex differences were evident. Feed efficiency metrics improved as birth BW increased ( < 0.05) except for RGR, where the contrary was observed. Correlations between most growth and feed efficiency metrics were strong to moderate ( < 0.05). Low-REI pigs (i.e., more efficient) had lower EI and ECR and were superior for RIG ( < 0.001) compared with high- and medium-REI pigs. High-RDG pigs (i.e., more efficient) had greater BW gain and better ECR ( < 0.001) compared with medium- and low-RDG pigs. Residual EI and RIG were both superior ( < 0.001) in high-RDG pigs compared with medium- and low-RDG pigs. Energy conversion ratio, REI, and RIG were superior ( < 0.05) in high-RMW pigs (i.e., more efficient) compared with medium-RMW pigs. High-RIG pigs (i.e., more efficient) had lower EI ( < 0.01) and superior ECR for RDG and REI compared with medium- and low-RIG pigs. In general, most of the correlations among the feed efficiency traits investigated in this study were different from unity, indicating that each trait is depicting a different aspect of efficiency in pigs, although the moderate to strong correlations suggest that improvement in one trait would, on average, lead to improvements in the others. Pigs ranked as more efficient on residual traits such as REI consumed less energy for a similar BW gain, which would translate into an economic benefit for pig producers.

Restoration of in situ fiber degradation and the role of fibrolytic microbes and ruminal pH in cows fed grain-rich diets transiently or continuously.

In this study, we used two different grain-rich feeding models (continuous or transient) to determine their effects on in situ fiber degradation and abundances of important rumen fibrolytic microbes in the rumen. The role of the magnitude of ruminal pH drop during grain feeding in the fiber degradation was also determined. The study was performed in eight rumen-fistulated dry cows. They were fed forage-only diet (baseline), and then challenged with a 60% concentrate diet for 4 weeks, either continuously (n=4 cows) or transiently (n=4 cows). The cows of transient feeding had 1 week off concentrate in between. Ruminal degradation of grass silage and fiber-rich hay was determined by the in situ technique, and microbial abundances attached to incubated samples were analyzed by quantitative PCR. The in situ trials were performed at the baseline and in the 1st and the last week of concentrate feeding in the continuous model. The in situ trials were done in cows of the transient model at the baseline and in the 1st week of the re-challenge with concentrate. In situ degradation of NDF and ADF of the forage samples, and microbial abundances were determined at 0, 4, 8, 24 and 48 h of the incubation. Ruminal pH and temperature during the incubation were recorded using indwelling pH sensors. Compared with the respective baseline, both grain-rich feeding models lowered ruminal pH and increased the duration of pH below 5.5 and 5.8. Results of the grass silage incubation showed that in the continuous model the extent of NDF and ADF degradation was lower in the 1st, but not in the last week compared with the baseline. For the transient model, degradation of NDF of the silage was lower during the re-challenge compared with the baseline. Degradation of NDF and ADF of the hay was suppressed by both feeding models compared with the respective baseline. Changes in fiber degradation of either grass silage or hay were not related to the magnitude of ruminal pH depression during grain-rich feeding. In both feeding models total fungal numbers and relative abundance of Butyrivibrio fibrisolvens attached to the incubated forages were decreased by the challenge. Overall, Fibrobacter succinogenes was more sensitive to the grain challenge compared with Ruminococcus albus and Ruminococcus flavefaciens. The study provided evidence for a restored ruminal fiber degradation after prolonged time of grain-rich feeding, however depending on physical and chemical characteristics of forages.

Comparison of electrogenic glucose transport processes and permeability between proximal and distal jejunum of laying hens.

1. The current objective was to assess (1) differences in mucosal transepithelial short-circuit current (Isc) and tissue conductance (GT), (2) the effect of a glucose stimulus and (3) epithelial paracellular permeability in the proximal and distal jejunum of laying hens. 2. Proximal and distal jejunal segments used in the Ussing chambers were collected at 9 ± 0.5 and 73 ± 3.4% (SEM) of jejunal length, respectively. The proximal jejunal mucosa showed a small negative Isc (-1.3 µA/cm2), whereas the distal jejunum had a higher Isc (32.9 µA/cm2). Similarly, GT was 2.5-fold greater in the distal compared to the proximal jejunum. 3. Increased paracellular permeability in the distal jejunum was displayed as demonstrated by a 5-fold higher mucosal to serosal flux of fluorescein isothiocyanate and horseradish peroxidase, representing molecules of low and high molecular weight, respectively. 4. Addition of glucose to the mucosal side (5 mmol/l, final concentration in the chamber) to stimulate an absorptive effect caused 3-fold greater GT in the distal compared to the proximal jejunum. 5. In conclusion, the present results supported site-specific electrogenic transport processes for the jejunal mucosa of laying hens. Therefore, precise description of the jejunal site may contribute to an improved comparability of electrophysiological data.

Epimural bacterial community structure in the rumen of Holstein cows with different responses to a long-term subacute ruminal acidosis diet challenge.

Subacute ruminal acidosis (SARA) is a prevalent metabolic disorder in cattle, characterized by intermittent drops in ruminal pH. This study investigated the effect of a gradual adaptation and continuously induced long-term SARA challenge diet on the epimural bacterial community structure in the rumen of cows. Eight rumen-cannulated nonlactating Holstein cows were transitioned over 1 wk from a forage-based baseline feeding diet (grass silage-hay mix) to a SARA challenge diet, which they were fed for 4 wk. The SARA challenge diet consisted of 60% concentrates (dry matter basis) and 40% grass silage-hay mix. Rumen papillae biopsies were taken at the baseline, on the last day of the 1-wk adaptation, and on the last day of the 4-wk SARA challenge period; ruminal pH was measured using wireless sensors. We isolated DNA from papillae samples for 16S rRNA gene amplicon sequencing using Illumina MiSeq. Sequencing results of most abundant key phylotypes were confirmed by quantitative PCR. Although they were fed similar amounts of concentrate, cows responded differently in terms of ruminal pH during the SARA feeding challenge. Cows were therefore classified as responders (n = 4) and nonresponders (n = 4): only responders met the SARA criterion of a ruminal pH drop below 5.8 for longer than 330 min/d. Data showed that Proteobacteria, Firmicutes, and Bacteroidetes were the most abundant phyla, and at genus level, Campylobacter and Kingella showed highest relative abundance, at 15.5 and 7.8%, respectively. Diversity analyses revealed a significant increase of diversity after the 1-wk adaptation but a decrease of diversity and species richness after the 4-wk SARA feeding challenge, although without distinction between responders and nonresponders. At the level of the operational taxonomic unit, we detected diet-specific shifts in epimural community structure, but in the overall epimural bacterial community structure, we found no differences between responders and nonresponders. Correlation analysis revealed significant associations between grain intake and operational taxonomic unit abundance. The study revealed major shifts in the 3 dominating phyla and, most importantly, a loss of diversity in the epimural bacterial communities during a long-term SARA diet challenge, in which 60% concentrate supply for 4 wk was instrumental rather than the magnitude of the drop of ruminal pH below 5.8.

Enzymatically modified starch up-regulates expression of incretins and sodium-coupled monocarboxylate transporter in jejunum of growing pigs.

Dietary effects on the host are mediated via modulation of the intestinal mucosal responses. The present study investigated the effect of an enzymatically modified starch (EMS) product on the mucosal expression of genes related to starch digestion, sugar and short-chain fatty acid (SCFA) absorption and incretins in the jejunum and cecum in growing pigs. Moreover, the impact of the EMS on hepatic expression of genes related to glucose and lipid metabolism, and postprandial serum metabolites were assessed. Barrows (n=12/diet; initial BW 29 kg) were individually fed three times daily with free access to a diet containing either EMS or waxy corn starch as control (CON) for 10 days. The enzymatic modification led to twice as many α-1,6-glycosidic bonds (~8%) in the amylopectin fraction in the EMS in comparison with the non-modified native waxy corn starch (4% α-1,6-glycosidic bonds). Linear discriminant analysis revealed distinct clustering of mucosal gene expression for EMS and CON diets in jejunum. Compared with the CON diet, the EMS intake up-regulated jejunal expression of sodium-coupled monocarboxylate transporter (SMCT), glucagon-like peptide-1 (GLP1) and gastric inhibitory polypeptide (GIP) (P<0.05) and intestinal alkaline phosphatase (ALPI) (P=0.08), which may be related to greater luminal SCFA availability, whereas cecal gene expression was unaffected by diet. Hepatic peroxisome proliferator-activated receptor γ (PPARγ) expression tended (P=0.07) to be down-regulated in pigs fed the EMS diet compared with pigs fed the CON diet, which may explain the trend (P=0.08) of 30% decrease in serum triglycerides in pigs fed the EMS diet. Furthermore, pigs fed the EMS diet had a 50% higher (P=0.03) serum urea concentration than pigs fed the CON diet potentially indicating an increased use of glucogenic amino acids for energy acquisition in these pigs. Present findings suggested the jejunum as the target site to influence the intestinal epithelium and altered lipid and carbohydrate metabolism by EMS feeding.

Enzymatically Modified Starch Favorably Modulated Intestinal Transit Time and Hindgut Fermentation in Growing Pigs.

Aside from being used as stabilizing agents in many processed foods, chemically modified starches may act as functional dietary ingredients. Therefore, development of chemically modified starches that are less digestible in the upper intestinal segments and promote fermentation in the hindgut receives considerable attention. This study aimed to investigate the impact of an enzymatically modified starch (EMS) on nutrient flow, passage rate, and bacterial activity at ileal and post-ileal level. Eight ileal-cannulated growing pigs were fed 2 diets containing 72% purified starch (EMS or waxy cornstarch as control) in a cross-over design for 10 d, followed by a 4-d collection of feces and 2-d collection of ileal digesta. On d 17, solid and liquid phase markers were added to the diet to determine ileal digesta flow for 8 h after feeding. Reduced small intestinal digestion after the consumption of the EMS diet was indicated by a 10%-increase in ileal flow and fecal excretion of dry matter and energy compared to the control diet (P<0.05). Moreover, EMS feeding reduced ileal transit time of both liquid and solid fractions compared to the control diet (P<0.05). The greater substrate flow to the large intestine with the EMS diet increased the concentrations of total and individual short-chain fatty acids (SCFA) in feces (P<0.05). Total bacterial 16S rRNA gene abundance was not affected by diet, whereas the relative abundance of the Lactobacillus group decreased (P<0.01) by 50% and of Enterobacteriaceae tended (P<0.1) to increase by 20% in ileal digesta with the EMS diet compared to the control diet. In conclusion, EMS appears to resemble a slowly digestible starch by reducing intestinal transit and increasing SCFA in the distal large intestine.

Comparison of growth performance and excreta composition in broiler chickens when ranked according to various feed efficiency metrics.

Several feed efficiency (FE) metrics are currently used in livestock production to select for improved FE. Whether or not different FE metrics similarly estimate physiological characteristics in chickens of diverging FE has not been reported so far. This study aimed to assess potential differences in feed intake (FI), performance, and nutrient excretion in broiler chickens of diverging FE when ranked according to their residual FI (RFI), residual BW gain (RBG), RFI and BW gain (RIG), and G:F between d 7 and 35 of life. The FI was determined daily and BW was recorded once a week. The ranking of chickens into good, medium, and poor FE groups was completed separately for each FE metric. Freshly dropped excreta were collected for pH and DM measurements on d 30 to 32 of life and total excreta for determination of nutrient excretion was collected on d 34 to 36 of life. Relationships among FE metrics were evaluated using regression analysis showing that RFI, RIG, and G:F were more related to each other than to RBG. The FE values greatly varied among chickens for all FE metrics and chickens did not always cluster within the same FE group when using RFI, RIG, RBG, and G:F as the FE metrics because of the calculation approaches. Due to sex-related differences in performance, data of male and female chickens were analyzed separately. The RFI and RIG metrics showed a linear increase ( < 0.01) in total FI from good to poor FE in male and female chickens, whereas G:F showed this effect ( ≤ 0.011) only when BW gain was standardized to 1,500 g. The RBG did not clearly select chickens of enhanced total BW gain and only tended ( < 0.1) to select for greater BW gain from good to poor FE in female chickens. Excreta pH linearly decreased by 0.7 log units and DM content increased in males from good to poor FE when using RFI and RIG, respectively ( < 0.01). In both sexes, RFI ( < 0.05) and RIG metrics ( ≤ 0.06) showed a linear increase in daily nitrogen excretion from good to poor FE. In conclusion, results demonstrate that selection of the metric used to determine the FE of chickens modified the results obtained for comparison of production parameters and nutrient excretion among FE groups. Thereby, the RFI, RIG, and G:F metrics were beneficial in selecting the most feed efficient chickens to reduce feed costs, whereas the use of RFI and RIG may be better to select chickens with improved nitrogen retention and thus reduced excretion of an environmental pollutant.

Transient feeding of a concentrate-rich diet increases the severity of subacute ruminal acidosis in dairy cattle.

The objective of this study was to investigate the effect of the pattern of concentrate-rich feeding on subacute ruminal acidosis (SARA), its severity, and the corresponding changes in VFA concentration. Eight rumen-cannulated Holstein cows were assigned to a 2 × 2 crossover design with 2 SARA challenge models and 2 experimental runs ( = 8 per treatment). Each run lasted for 40 d, consisting of a 6-d baseline, a 6-d gradual grain adaptation, and a 28-d SARA challenge period. The 2 SARA challenge models were transient (TRA) and persistent (PER) SARA. Initially, all cows were subjected to a forage-only diet (baseline) and gradually switched to 60% concentrate (DM basis). Then, cows in the PER model were continuously challenged for 28 d, whereas cows in the TRA model had a 7-d break from the SARA diet and were fed the forage-only diet after the first 7 d of SARA challenge. Thereafter, the TRA cows were rechallenged with the SARA diet. Wireless ruminal pH sensors were used to obtain ruminal pH profiles and temperature over the experimental period. For the determination of VFA, free ruminal liquid (FRL) and particle-associated ruminal liquid (PARL) were collected once for the baseline and twice (d 20 and 40 for the PER model) or 3 times (d 13, 30, and 40 for the TRA model) during SARA, each time at 0, 4, and 8 h after the morning feeding. Cows in both models experienced SARA albeit with day-to-day variation. From the start until the first 7-d SARA, cows of both models had similar pH profiles, but during the rechallenge, SARA was more severe in the TRA model than in the PER model based on lower daily mean ruminal pH (5.93 vs. 6.15; SEM 0.058) and double the amount of time at pH < 5.8 (497 vs. 278 min; SEM 68.61, < 0.05). Mean ruminal temperature was raised during SARA compared with the baseline (38.9 vs. 38.7°C; SEM 0.057, < 0.001). Concentrations of VFA increased with increasing time after feeding ( < 0.001). In general, SARA challenge (d 40 vs. the baseline), but not the challenge model, altered VFA concentrations and profile of both FRL and PARL by increasing the amounts of propionate and butyrate, whereas total VFA concentration was less affected. Proportions of VFA shifted over the duration of SARA challenge with more propionate but less acetate and butyrate proportions with advancing days of SARA challenge, leading to the values of the last SARA day being different from the earlier days ( < 0.05). In conclusion, the TRA condition led to the higher severity of SARA, but factors beyond feed intake and VFA alterations seemed to play a role.

Nutrition, rumen health and inflammation in the transition period and their role on overall health and fertility in dairy cows.

Transition is a stressful period and critical for the entire cow's productive lifespan and reproduction. Optimal feeding management during transition period enables smooth metabolic adaptation to the initiation of lactation. Major nutritional challenge during this period is the urgent need to counteract the drastic deficits in energy and nutrients of the early-lactating cow. This is primarily done by inclusion of large amounts of concentrates in the diet during early lactation, causing major dietary imbalances with utmost importance for rumen health. Proper feeding management targeting rumen health in the transition period improves nutrient degradation and the net supply with energy and key nutrients of the host while preventing systemic disturbances and inflammation, events which are instrumental for cow's overall health and reproductive performance. The review provides insights into the role of, and gives practical hints regarding diet balancing efforts and feeding management strategies targeting rumen health and systemic inflammation during the periparturient period with the aim to enhance cow health and fertility.

Microbial populations and fermentation profiles in rumen liquid and solids of Holstein cows respond differently to dietary barley processing.

AIMS: To evaluate the effects of treating barley grain with lactic acid (LA) and heat on postprandial dynamics of 19 microbial taxa and fermentation in the rumen of dairy cows. METHODS AND RESULTS: This study was designed as a double 3 × 3 Latin square with six rumen-cannulated cows and three diets either containing untreated control barley or barley treated with 1% LA and 1% LA and heat (LAH, 55°C). Microbial populations, pH and volatile fatty acids were assessed in rumen liquid and solids during the postprandial period. Propionate increased and butyrate decreased in rumen solids of cows fed LA and LAH treated barley compared to the control barley. The LA but not LAH treatment depressed Fibrobacter succinogenes in rumen liquid and solids, whereas the opposite effect was observed for Ruminococcus albus in both fractions and Ruminococcus flavefaciens in rumen solids. LA promoted Ruminobacter amylophilus with the effect being more pronounced with LAH. The Lactobacillus group and Megasphaera elsdenii increased in both fractions with LA but not with LAH. CONCLUSIONS: LA and LAH treatment of barley differently altered ruminal abundance of certain bacterial taxa and fungi and increased propionate fermentation in rumen solids, whereby LA and LAH effects were consistent and mostly independent of the rumen fraction and time after barley feeding. SIGNIFICANCE AND IMPACT OF THE STUDY: Results provided evidence that LA and LAH treatment of barley can enhance rumen propionate fermentation without adversely affecting rumen pH. As propionate is the major contributor to gluconeogenesis in ruminants, the present barley treatment may have practical application to enhance energy supply in dairy cows.

High diversity of viable bacteria isolated from lymph nodes of slaughter pigs and its possible impacts for food safety.

AIMS: Ileocaecal lymph nodes (ICLNs) of pigs, the key immune inductive site for bacterial systemic invasion, were examined in this study with emphasis on viable and cultivable bacteria. METHODS AND RESULTS: Asymptomatic and pathologically altered ICLNs of slaughter pigs (n = 16) with hyperplasia, purulence or granulomatous formations were aerobically and anaerobically cultivated. In total, 209 isolates were collected and the near full-length 16S rRNA gene from each isolate was sequenced. Taxonomic classification revealed that 68% of the isolates belonged to Proteobacteria, 27% to Firmicutes and 5% to Actinobacteria. Purulent and granulomatous ICLNs generally tended to contain more Proteobacteria than asymptomatic and enlarged ICLNs (P = 0·061). The isolates could be assigned to 25 species belonging to 17 genera including Escherichia, Carnobacterium, Lactobacillus, Staphylococcus and Acinetobacter. Furthermore, pathogens such as Streptococcus suis and Salmonella enterica were detected. The most abundant isolate (57%) was most similar (>99%) to Escherichia coli. Random amplified polymorphic DNA (RAPD) profiling revealed a high genetic diversity among E. coli isolates and 24% of these isolates were positive for at least one gene associated with enterohemorrhagic disease (eae, fliC, stx1 or hlyA). Compared with a recently published DNA-based high-throughput sequencing data set including the same ICLNs, 4% of species detected were cultivable. CONCLUSIONS: The presence of viable, commensal and pathogenic bacterial phylotypes could be proven in ICLNs with Proteobacteria being dominant. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study that broadly characterizes viable bacteria from ICLNs of pigs. The presence of bacteria in lymph nodes of farm animals has practical relevance for host colonization and possible chronic infection. It is also of great interest for basic research investigating translocation of bacteria from the gastrointestinal tract to ICLNs.

Treatment of grain with organic acids at 2 different dietary phosphorus levels modulates ruminal microbial community structure and fermentation patterns in vitro.

Recent data indicate positive effects of treating grain with citric (CAc) or lactic acid (LAc) on the hydrolysis of phytate phosphorus (P) and fermentation products of the grain. This study used a semicontinuous rumen simulation technique to evaluate the effects of processing of barley with 50.25 g/L (wt/vol) CAc or 76.25 g/L LAc on microbial composition, metabolic fermentation profile, and nutrient degradation at low or high dietary P supply. The low P diet [3.1g of P per kg of dry matter (DM) of dietary P sources only] was not supplemented with inorganic P, whereas the high P diet was supplemented with 0.5 g of inorganic P per kg of DM through mineral premix and 870 mg of inorganic P/d per incubation fermenter via artificial saliva. Target microbes were determined using quantitative PCR. Data showed depression of total bacteria but not of total protozoa or short-chain fatty acid (SCFA) concentration with the low P diet. In addition, the low P diet lowered the relative abundance of Ruminococcus albus and decreased neutral detergent fiber (NDF) degradation and acetate proportion, but increased the abundance of several predominantly noncellulolytic bacterial species and anaerobic fungi. Treatment of grain with LAc increased the abundance of total bacteria in the low P diet only, and this effect was associated with a greater concentration of SCFA in the ruminal fluid. Interestingly, in the low P diet, CAc treatment of barley increased the most prevalent bacterial group, the genus Prevotella, in ruminal fluid and increased NDF degradation to the same extent as did inorganic P supplementation in the high P diet. Treatment with either CAc or LAc lowered the abundance of Megasphaera elsdenii but only in the low P diet. On the other hand, CAc treatment increased the proportion of acetate in the low P diet, whereas LAc treatment decreased this variable at both dietary P levels. The propionate proportion was significantly increased by LAc at both P levels, whereas butyrate increased only with the low P diet. Treatments with CAc or LAc reduced the degradation of CP and ammonia concentration compared with the control diet at both P levels. In conclusion, the beneficial effects of CAc and LAc treatment on specific ruminal microbes, fermentation profile, and fiber degradation in the low P diet suggest the potential for the treatment to compensate for the lack of inorganic P supplementation in vitro. Further research is warranted to determine the extent to which the treatment can alleviate the shortage of inorganic P supplementation under in vivo conditions.

Pyrosequencing reveals shifts in the bacterial epimural community relative to dietary concentrate amount in goats.

Ecological balance in the rumen is highly sensitive to concentrate-rich diets. Yet the effects of these feeding practices on the caprine bacterial epimural microbiome (CBEM), a microbial community with putative important physiological functions in the rumen, are largely unexplored. This study aimed to investigate the effect of dietary concentrate amount on ruminal CBEM. Seventeen growing goats were fed diets with 0 [n=5; 6.2MJ of metabolizable energy (ME)/d], 30 (n=6; 7.3MJ of /d), or 60% (n=6; 10.2MJ of ME/d) concentrate for 6 wk. Two hours after their last feeding, goats were euthanized and tissue samples of the ventral rumen wall were collected, washed in phosphate-buffered saline to detach loosely attached bacteria, and stored at -20°C for further processing. Genomic DNA was isolated from thawed rumen mucosa samples and used for Roche/454 Life Science (Branford, CT) 16S rRNA gene amplicon pyrosequencing yielding 122,458 reads. Pyrosequencing data were clustered into 1,879 operational taxonomic units (OTU; 0.03 distance level). Pyrosequencing revealed Proteobacteria, Bacteroidetes, Firmicutes, and Spirochaetes as the most abundant phyla (97.7%). Compared with the 30% group, both the 60 and 0% concentrate groups harbored significantly more Firmicutes and SR1, respectively. On an OTU level, a Bergeriella-related OTU was most abundant in the CBEM, followed by 2 Campylobacter OTU, which responded differently to diets: 1 OTU was significantly increased whereas the other significantly decreased with highest concentrate amount in the diet. At the genus level, the 0% concentrate group harbored increased Kingella-like sequences compared with the other feeding groups. Furthermore, the 0% concentrate group tended to have more Bergeriella than the 30 and 60% concentrate groups. The genus Bergeriella was significantly decreased in the 60% feeding group compared with the other diets. In conclusion, this is the first report of CBEM using deep-sequencing methods on the genus and OTU level, and our study revealed major shifts in the CBEM in response to concentrate-rich diets with potential health relevance in goats.

Substitution of common concentrates with by-products modulated ruminal fermentation, nutrient degradation, and microbial community composition in vitro.

A rumen simulation technique was used to evaluate the effects of the complete substitution of a common concentrate mixture (CON) with a mixture consisting solely of by-products from the food industry (BP) at 2 different forage-to-concentrate ratios on ruminal fermentation profile, nutrient degradation, and abundance of rumen microbiota. The experiment was a 2×2 factorial arrangement with 2 concentrate types (CON and BP) and 2 concentrate levels (25 and 50% of diet dry matter). The experiment consisted of 2 experimental runs with 12 fermentation vessels each (n=6 per treatment). Each run lasted for 10d, with data collection on the last 5d. The BP diets had lower starch, but higher neutral detergent fiber (NDF) and fat contents compared with CON. Degradation of crude protein was decreased, but NDF and nonfiber carbohydrate degradation were higher for the BP diets. At the 50% concentrate level, organic matter degradation tended to be lower for BP and CH4 formation per unit of NDF degraded was also lower for BP. The BP mixture led to a higher concentration of propionate and a lower acetate-to-propionate ratio, whereas concentrations of butyrate and caproate decreased. Concentrate type did not affect microbial community composition, except that the abundance of bacteria of the genus Prevotella was higher for BP. Increasing the concentrate level resulted in higher degradation of organic matter and crude protein. At the higher concentrate level, total short-chain fatty acid formation increased and concentrations of isobutyrate and valerate decreased. In addition, at the 50% concentrate level, numbers of protozoa increased, whereas numbers of methanogens, anaerobic fungi, and fibrolytic bacteria decreased. No interaction was noted between the 2 dietary factors on most variables, except that at the higher concentrate level the effects of BP on CH4 and CO2 formation per unit of NDF degraded, crude protein degradation, and the abundance of Prevotella were more prominent. In conclusion, the results of this study suggest that BP in the diet can adequately substitute CON with regard to ruminal fermentation profile and microbiota, showing even favorable fermentation patterns when fed at 50% inclusion rate.