M. Hollmann

Feeding barley grain-rich diets altered electrophysiological properties and permeability of the ruminal wall in a goat model

High-producing ruminants are commonly fed large amounts of concentrate to meet their high energy demands for rapid growth or high milk production. However, this feeding strategy can severely impair rumen functioning, leading to subacute ruminal acidosis. Subacute ruminal acidosis might have consequences for electrophysiological properties by changing the net ion transfer and permeability of ruminal epithelia, which may increase the uptake of toxic compounds generated in the rumen into the systemic circulation. The objective of the present study was to investigate the effects of excessive barley feeding on the electrophysiological and barrier functions of the ruminal epithelium and serum inflammation and ketogenesis markers after a long-term feeding challenge, using growing goats as a ruminant model. A feeding trial was carried out with growing goats allocated to 1 of the 3 groups (n=5-6 animals/group), with diets consisting exclusively of hay (control diet) or hay with 30 or 60% barley grain. Samples of the ventral ruminal epithelium were taken after euthanasia and instantly subjected to Ussing chamber experiments, where electrophysiological properties of the epithelium were measured in parallel with the permeability of marker molecules of different sizes [fluorescein 5(6)-isothiocyanate and horseradish peroxidase] from luminal to apical side. Additionally, ruminal fluid and blood samples were taken at the beginning of the experiment as well as shortly before euthanasia. Ruminal fluid samples were analyzed for volatile fatty acids and pH, whereas blood samples were analyzed for lipopolysaccharide, serum amyloid A, and β-hydroxybutyrate. Electrophysiological data indicated that barley feeding increased the epithelial short-circuit current compared with the control. Tissue conductance also increased with dietary barley inclusion. As shown with both marker molecules, permeability of ruminal epithelia increased with barley inclusion in the diet. Despite a lowered ruminal pH associated with increased volatile fatty acids (such as propionate and butyrate) concentrations as well as altered epithelial properties in response to high-grain feeding, no signs of inflammation became apparent, as blood serum amyloid A concentrations remained unaffected by diet. However, greater amounts of grain in the diet were associated with a quadratic increase in lipopolysaccharide concentration in the serum. Also, increasing the amounts of barley grain in the diet resulted in a tendency to quadratically augment serum concentrations of β-hydroxybutyrate and, hence, the alimentary ketogenesis. Further studies are needed to clarify the role of barley inclusion in the development of subacute ruminal acidosis in relation to ruminal epithelial damage and the translocation of toxic compounds in vivo.

Epithelial response to high-grain diets involves alteration in nutrient transporters and Na+/K+-ATPase mRNA expression in rumen and colon of goats1

Emerging evidence at the mRNA level indicates that feeding high-grain diets to ruminants leads to coordinated changes in the molecular response of the rumen epithelium. Yet, epithelial adaptation of the hindgut to increasing dietary grain levels has not been established in ruminants. Therefore, the objective of this study was to characterize alterations in mRNA expression associated with nutrient transport and electrochemical gradients in rumen and colon epithelium, and rumen morphology in growing goats fed different grain levels. Goats (n = 6) were fed diets with increasing levels of 0, 30, or 60% barley grain for 6 wk. Goats were euthanized 2 h after their last feeding, and digesta and tissue samples of the cranial part of the ventral rumen and proximal colon were collected. Goats fed the 60% grain diet exhibited a lower ruminal and colonic pH (P < 0.01) and a greater colonic total VFA concentration (P < 0.05) compared with those fed the 0 and 30% grain diets. As response to the decreased ruminal pH, goats fed the 60% grain diet had a greater (P < 0.05) keratinization and thicker stratum corneum of the rumen epithelium than goats fed the 0 and 30% grain diets. The 60% grain diet upregulated (P < 0.05) MCT1 expression by 45% and downregulated (P < 0.05) the expression of MCT4 and SGLT1 by 28 and 50%, respectively, in rumen epithelium compared with the 0 and 30% grain diets. Accordingly, goats fed the 60% grain diet had a greater (P < 0.05) expression of MCT1 and ATP1A1 in colon epithelium than goats fed the 0 and 30% grain diets. Regression analyses showed negative relationships (R(2) = 0.35 to 0.87, P < 0.05) of MCT1 and ATP1A1 expression in rumen and colon epithelium and thickening of ruminal stratum corneum to decreasing luminal pH values, suggesting greater mRNA expression at lower pH. In contrast, MCT4 expression in rumen epithelium positively correlated to luminal pH (R(2) = 0.95, P < 0.01). In conclusion, results of this model study indicated that with the greatest grain level rumen and colon molecular epithelial responses may have been related to counteract the consequences of luminal acidification on intracellular homeostasis in epithelial cells and concomitantly to increase systemic absorption of VFA.

Downregulation of Cellular Protective Factors of Rumen Epithelium in Goats Fed High Energy Diet

Energy-rich diets can challenge metabolic and protective functions of the rumen epithelial cells, but the underlying factors are unclear. This study sought to evaluate proteomic changes of the rumen epithelium in goats fed a low, medium, or high energy diet. Expression of protein changes were compared by two-dimensional differential gel electrophoresis followed by protein identification with matrix assisted laser desorption ionisation tandem time-of-flight mass spectrometry. Of about 2,000 spots commonly detected in all gels, 64 spots were significantly regulated, which were traced back to 24 unique proteins. Interestingly, the expression profiles of several chaperone proteins with important cellular protective functions such as heat shock cognate 71 kDa protein, peroxiredoxin-6, serpin H1, protein disulfide-isomerase, and selenium-binding protein were collectively downregulated in response to high dietary energy supply. Similar regulation patterns were obtained for some other proteins involved in transport or metabolic functions. In contrast, metabolic enzymes like retinal dehydrogenase 1 and ATP synthase subunit beta, mitochondrial precursor were upregulated in response to high energy diet. Lower expressions of chaperone proteins in the rumen epithelial cells in response to high energy supply may suggest that these cells were less protected against the potentially harmful rumen toxic compounds, which might have consequences for rumen and systemic health. Our findings also suggest that energy-rich diets and the resulting acidotic insult may render rumen epithelial cells more vulnerable to cellular damage by attenuating their cell defense system, hence facilitating the impairment of rumen barrier function, typically observed in energy-rich fed ruminants.

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.

Detection of 3-nitropropionic acid and cytotoxicity inMucor circinelloides.

Mucorales are regarded as the aetiological agents of Mucormycosis. Their capabilities to produce mycotoxins are not profoundly investigated, in contrast to those of the fungi from the generaPenicillium, Aspergillus, orFusarium. The aim of this study was to isolate and identify fungi of the order Mucorales and investigate mycotoxins production. Twelve samples of visibly moulded grass silage and eight samples of damaged whole crop maize silage were analysed. Malt extract agar plates were used for sub cultivation. Three fungal species of the order Mucorales were isolated from grass silage, which were identified by their macro-and micro-morphology asAbsidia corymbifera, Mucor circinelloides andRhizopus stolonifer. The cytotoxicity ofMucor circinelloides extract was analysed using the cytotoxicity test (MTT assay) and the result, showed a low cytotoxicity. Additionally extracts fromAbsidia corymbifera, Mucor circinelloides andRhizopus stolonifer were tested for mycotoxin-production using an LC/MS/MS-based multimycotoxin method. 3-nitropropionic acid was detected in the culture extract ofMucor circinelloides.

Finishing pigs that are divergent in feed efficiency show small differences in intestinal functionality and structure

Controversial information is available regarding the feed efficiency-related variation in intestinal size, structure and functionality in pigs. The present objective was therefore to investigate the differences in visceral organ size, intestinal morphology, mucosal enzyme activity, intestinal integrity and related gene expression in low and high RFI pigs which were reared at three different geographical locations (Austria, AT; Northern Ireland, NI; Republic of Ireland, ROI) using similar protocols. Pigs (n = 369) were ranked for their RFI between days 42 and 91 postweaning and low and high RFI pigs (n = 16 from AT, n = 24 from NI, and n = 60 from ROI) were selected. Pigs were sacrificed and sampled on ~day 110 of life. In general, RFI-related variation in intestinal size, structure and function was small. Some energy saving mechanisms and enhanced digestive and absorptive capacity were indicated in low versus high RFI pigs by shorter crypts, higher duodenal lactase and maltase activity and greater mucosal permeability (P < 0.05), but differences were mainly seen in pigs from AT and to a lesser degree in pigs from ROI. Additionally, low RFI pigs from AT had more goblet cells in duodenum but fewer in jejunum compared to high RFI pigs (P < 0.05). Together with the lower expression of TLR4 and TNFA in low versus high RFI pigs from AT and ROI (P < 0.05), these results might indicate differences in the innate immune response between low and high RFI pigs. Results demonstrated that the variation in the size of visceral organs and intestinal structure and functionality was greater between geographic location (local environmental factors) than between RFI ranks of pigs. In conclusion, present results support previous findings that the intestinal size, structure and functionality do not significantly contribute to variation in RFI of pigs.

Aflatoxins in selected Thai commodities

Aflatoxin (AF) B1, B2, G1 and G2 were determined in 120 samples of selected Thai commodities including unpolished rice, unpolished glutinous rice, chilli powder, whole dried chilli pods and raw peanut. The mean concentrations of the total AFs for analysed samples were 0.16, 25.43, 14.18, 6.62 and 1.43 µg kg−1 with positive incidences of 4%, 20%, 97%, 37% and 30%, respectively. Quantitative analysis was performed using HPLC equipped with post-column derivatisation and fluorescence detection. Sample clean-up was carried out using immunoaffinity columns for selective enrichment of AFs. The method was validated by using certified reference material, which showed recoveries over 85%. The limit of detections (LODs) and limit of quantifications (LOQs) were in a range between 0.01–0.11 µg kg−1 and 0.03–0.38 µg kg−1, respectively. The results clearly demonstrated that AFs were detectable in different matrices. Chilli powder was found to have the highest level of AFs contamination followed by chilli pods, peanut and rice, respectively. However, among the selected commodities, unpolished rice contained only trace levels of AFB1 and AFB2. With regard to the fact that AFs are a natural contaminant in commodities, this report calls to attention the regular monitoring and effective control of food commodities to prevent health hazards.

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

ABSTRACT 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.

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

ABSTRACT 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.

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

Abstract 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.