Effects of selenium enrichment on fermentation characteristics, selenium content and microbial community of alfalfa silage.

BACKGROUND: Selenium is essential for livestock and human health. The traditional way of adding selenium to livestock diets has limitations, and there is a growing trend to provide livestock with a safe and efficient source of selenium through selenium-enriched pasture. Therefore, this study was conducted to investigate the effects of selenium enrichment on fermentation characteristics, selenium content, selenium morphology, microbial community and in vitro digestion of silage alfalfa by using unenriched (CK) and selenium-enriched (Se) alfalfa as raw material for silage. RESULTS: In this study, selenium enrichment significantly increased crude protein, soluble carbohydrate, total selenium, and organic selenium contents of alfalfa silage fresh and post-silage samples, and it significantly decreased neutral detergent fiber and acid detergent fiber contents (p < 0.05). Selenium enrichment altered the form of selenium in plants, mainly in the form of SeMet and SeMeCys, which were significantly higher than that of CK (p < 0.05). Selenium enrichment could significantly increase the lactic acid content, reduce the pH value, change the diversity of bacterial community, promote the growth of beneficial bacteria such as Lactiplantibacillus and inhibit the growth of harmful bacteria such as Pantoea, so as to improve the fermentation quality of silage. The in vitro digestibility of dry matter (IVDMD), in vitro digestibility of acid detergent fibers (IVADFD) and in vitro digestibility of acid detergent fibers (IVNDFD) of silage after selenium enrichment were significantly higher than those of CK (p < 0.05). CONCLUSION: This study showed that the presence of selenium could regulate the structure of the alfalfa silage bacterial community and improve alfalfa silage fermentation quality. Selenium enrichment measures can change the morphology of selenium in alfalfa silage products, thus promoting the conversion of organic selenium.

Development and application of a multi-step porcine in vitro system to evaluate feedstuffs and feed additives for their efficacy in nutrient digestion, digesta characteristics, and intestinal immune responses.

In vitro model provides alternatives to the use of live animals in research. In pig nutrition, there has been a tremendous increase in in vivo research over the decades. Proper utilization of in vitro models could provide a screening tool to reduce the needs of in vivo studies, research duration, cost, and the use of animals and feeds. This study aimed to develop a multi-step porcine in vitro system to simulate nutrient digestion and intestinal epithelial immune responses affected by feedstuffs and feed additives. Seven feedstuffs (corn, corn distillers dried grains with solubles [corn DDGS], barley, wheat, soybean meal, soy protein concentrates, and Corynebacterium glutamicum cell mass [CGCM]), feed enzymes (xylanase and phytase), and supplemental amino acids (arginine, methionine, and tryptophan), were used in this in vitro evaluation for their efficacy on digestibility, digesta characteristics, and intestinal health compared with the results from previously published in vivo studies. All in vitro evaluations were triplicated. Data were analyzed using Mixed procedure of SAS9.4. Evaluations included (1) nutrient digestibility of feedstuffs, (2) the effects of feed enzymes, xylanase and phytase, on digestibility of feedstuffs and specific substrates, and (3) the effects of amino acids, arginine, tryptophan, and methionine, on anti-inflammatory, anti-oxidative, and anti-heat stress statuses showing their effects (P < 0.05) on the measured items. Differences in dry matter and crude protein digestibility among the feedstuffs as well as effects of xylanase and phytase were detected (P < 0.05), including xylo-oligosaccharide profiles and phosphorus release from phytate. Supplementation of arginine, tryptophan, and methionine modulated (P < 0.05) cellular inflammatory and oxidative stress responses. The use of this in vitro model allowed the use of 3 experimental replications providing sufficient statistical power at P < 0.05. This indicates in vitro models can have increased precision and consistency compared with in vivo animal studies.

Bioaccessibility and Antioxidant Capacity of Grape Seed and Grape Skin Phenolic Compounds After Simulated In Vitro Gastrointestinal Digestion.

Grapes present recognized beneficial effects on human health due to their polyphenolic composition. The grape overproduction together with the wine sales down and the world socioeconomic situation makes the wine grape valorization a promising strategy to give an added-value to this natural product. The objective of the present work was to study the influence of in vitro gastrointestinal digestion on antioxidant capacity and polyphenolic profile of skin and seed extracts of different grape varieties (Tempranillo, Graciano, Maturana tinta and Hondarrabi zuri). After in vitro gastrointestinal digestion, total phenolic content (TPC) of seed polyphenolic extracts decreased significantly for all the varieties. The highest decrease was for Tempranillo going from 108 ± 9 to 50 ± 3 mg / g dry matter (dm). This variety also showed the highest decrease of 90% in antioxidant capacity. However, for all the skin polyphenolic extracts there was an increase in TPC. The highest variation was also for Tempranillo. It varied from 10.1 ± 0.8 to 55.1 ± 0.9 mg / g dm. Among red varieties Tempranillo skin polyphenolic extract showed the lowest undigested anthocyanin content but the highest bioaccessibility index (BI) of 77%. For flavanols, flavonols and procyanidins the seed polyphenolic extracts showed a BI at the intestinal phase between 11% for (+)-epicatechin gallate to 130% procyanidin A2. The results of this study suggest that grape skin extracts and grape seed extracts are a reliable source of bioaccessible antioxidant polyphenols, to be used for the development of antioxidant supplements with specific functionalities depending on the grape variety.

Comparison of forages' digestion levels for different in vitro digestion techniques in horses.

BACKGROUND: Forages are widely used in horse diets. Different in vitro techniques are being tried to determine the fermentation levels of forages in the horse digestive tract. OBJECTIVES: This study aimed to evaluate the digestion levels of four dry forages commonly used in horse nutrition: alfalfa herbage, meadow hay, wheat straw, and Italian ryegrass. In vitro total digestion (TDT), in vitro Sunvold-large intestine digestion (SDT) and in vitro Menke-large intestine digestion (MDT) techniques were compared. METHODS: The study determined in vitro true dry matter digestion (T-DMD), in vitro true organic matter digestion (T-OMD) and in vitro true neutral detergent fibre digestion (T-NDFD). Additionally, concentrations of straight short-chain fatty acids (SCFAs; acetic acid - AA, propionic acid , butyric acid, and valeric acid ) and branched short-chain fatty acids (BSCFA) were assessed. RESULTS: The highest in vitro T-DMD, T-OMD and T-NDFD values were determined by the in vitro TDT for the four forages (p < 0.05). In vitro T-DMD and T-OMD values of alfalfa herbage were higher than those of Italian ryegrass, meadow hay and wheat straw in the in vitro TDT (p < 0.001). In addition, in vitro T-DMD and T-OMD values of alfalfa herbage in the in vitro SDT were higher than those of meadow hay and wheat straw (p < 0.001). In the in vitro TDT, the molarity of AA, total SCFA and BSCFA in the digestion fluid of alfalfa herbage was higher than those of other forages (p < 0.05). CONCLUSION: The in vitro total enzymatic + fermentative digestion technique for horse forages revealed higher values than the in vitro fermentative digestion techniques. In general, the higher the non-structural carbohydrate and crude protein contents in the forage, the higher the results of the in vitro TDT compared to the other techniques.

Characterization of ruminal degradation, intestinal digestion and total true nutrient supply to dairy cows from feedstocks and coproducts from Canola bio-oil processing: Impact by source origin.

The objective of this study was to characterize ruminal degradation, intestinal digestion and total true nutrient supply to dairy cows from canola feedstock (canola seeds) and coproducts (meal and pellets) from bio-oil processing which were impacted by source origin. The feedstocks and coproducts (mash, pellet) were randomly collected from five different bio-oil processing plants with five different batches of samples in each bio-processing plant in Canada (CA) and China (CH). In situ rumen degradation kinetics were determined using four fistulated Holstein cows with incubation times at 0, 2, 4, 8, 12, 24 and 48 h. Intestinal digestions were determined using the three-step in vitro method with preincubation at 12 h. The DVE/OEB and National Research Council systems were applied to evaluate the truly absorbable nutrient supply to dairy cows and feed milk values (FMVs). The results showed that in situ undegradable fractions (U) (p = 0.025) were higher in CA meals, and potentially degradable fraction of D was higher (p = 0.016) in CH meals. CH meals had higher total digestible dry matter (TDDM, p = 0.018) and intestinal digestibility of protein (dIDP, p = 0.016). Canola meals from CA had lower MREE (microbial protein synthesized in the rumen based on available rumen degradable protein; p = 0.011) and DVME (rumen synthesized microbial protein digested in the small intestine; p = 0.011) and had higher ECP (endogenous protein in the small intestine, p = 0.001) and absorbed endogenous crude protein (truly absorbed ECP in the small intestine) than CH (p = 0.001). The FMV evaluated based on the metabolic protein and net energy showed no differences between CA and CH in both coproducts and feedstocks.

Assessing the In Vitro and In Vivo Effect of Supplementation with a Garlic (Allium sativum) and Oregano (Origanum vulgare) Essential Oil Mixture on Digestibility in West African Sheep.

This study assessed the impact of a mixture of garlic (Allium sativum) and oregano (Origanum vulgare) essential oils (EOGOs) on in vitro dry matter digestibility (IVDMD) and in vivo apparent nutrient digestibility. Different EOGO inclusion levels were evaluated to assess the dose response and potential effects of the mixture. Three EOGO inclusion levels (0.5, 0.75, and 1 mL/kg of incubated dry matter) were evaluated in vitro, while four treatments (0.5, 0.75, and 1 mL/day of EOGO and a control group) were tested in vivo on 12 West African sheep. A randomized controlled trial was conducted using a 4 × 4 design. Blood parameters (glucose, blood urea nitrogen, and ß-hydroxybutyrate) were measured to observe the effect of EOGO on the metabolism. The results showed that the inclusion of EOGO significantly enhanced IVDMD at low levels (p < 0.052) compared with the highest levels in treatments containing 0.5 and 0.75 mL/kg of EOGO dry matter. A higher intake of dry matter (DM), crude protein (CP), and neutral detergent fiber (NDF) (p < 0.05) was observed in the in vivo diets with the inclusion of EOGO. In terms of in vivo apparent digestibility, significant differences were found among treatments in the digestibility coefficients of DM, CP, and NDF. EOGO inclusion increased the digestibility of DM. CP digestibility displayed a cubic effect (p < 0.038), with the lowest values of digestibility observed at 1 mL EOGO inclusion. Additionally, NDF digestibility showed a cubic effect (p < 0.012), with the highest value obtained at 0.75 mL of EOGO inclusion. The inclusion levels above 0.75 mL EOGO showed a cubic effect, which indicates that higher concentrations of EOGO may not be beneficial for the digestibility of CP and NDF. Although no significant difference was observed in total digestible nutrients, a linear trend was observed (p < 0.059). EOGO improved the intake of DM, CP, and NDF. EOGO supplementation improved the digestibility of DM and NDF, with optimal levels observed at 0.5 mL/day. No significant effects were observed in the blood parameters. These results suggest that EOGO has the potential as an additive in ruminal nutrition to improve food digestibility and serve as an alternative to antibiotic additives. The use of EOGO potentially improves fiber digestion and may reduce the use of antibiotics in livestock production. Garlic (A. sativum) and oregano (O. vulgare) essential oils effectively modulated fiber digestibility at 0.75 mL/day. Garlic (A. sativum) and oregano (O. vulgare) essential oils have the potential to improve digestibility at low inclusion levels and serve as an alternative to antibiotic additives. The effectiveness of essential oils is greater in a mixture and at lower doses.

Modulation of digestibility of canine food using enzyme supplement: an in vitro simulated semi-dynamic digestion study.

Digestibility and nutrient availability are important parameters when estimating the nutritional quality of pet food. We have developed a simulated semi-dynamic in vitro canine digestion model to evaluate the digestibility of dry extruded canine food. Canine food was assessed for digestible energy, dry matter digestibility, protein digestibility, non-fibrous carbohydrate (NFC) digestibility, and total antioxidant capacity (TAC) in the absence and presence of an enzyme blend (DigeSEB Super Pet). Enzyme blend supplementation in canine food was found to increase the dry matter digestibility (18.7%, p < 0.05), digestible energy (18.1%, p < 0.05), and protein digestibility (11%, p < 0.1) and reducing sugar release (106.3%, p < 0.005). The release of low molecular weight peptides (48.7%) and essential amino acids (15.6%) increased within 0.5 h of gastrointestinal digestion due to enzyme blend supplementation. Furthermore, the TAC of the digesta was also increased (8.1%, p < 0.005) in the canine food supplemented with enzyme blend. Overall, supplementation of enzyme blend in canine food is an effective strategy to enhance the food digestibility and nutrient availability for absorption.

Bioaccessibility of Tocols in Commercial Maize Hybrids Determined by an In Vitro Digestion Model for Poultry.

Despite the high proportion of maize grain in animal diets, the contribution made by maize phytochemicals is neglected. Tocols and their contribution to the vitamin E content of animal diets are one example, exacerbated by sparse information on the tocol bioaccessibility of commercial hybrids. In this study, the contents of individual and total tocols and their bioaccessibility were determined in the grain samples of 103 commercial hybrids using a standardized INFOGEST digestion procedure. In the studied hybrids, total tocol content ranged from 19.24 to 54.44 µg/g of dry matter. The contents of micellar α-, γ-, δ-tocopherols, γ-tocotrienol, and total tocols correlated positively with the corresponding contents in the grain samples of the studied hybrids. In contrast, a negative correlation was observed between the bioaccessibility of γ- tocopherol, α- and γ-tocotrienol, and total tocols, along with the corresponding contents in the grain of studied hybrids. The highest bioaccessibility was exhibited by γ-tocotrienol (532.77 g/kg), followed by δ-tocopherol (529.88 g/kg), γ-tocopherol (461.76 g/kg), α-tocopherol (406.49 g/kg), and α-tocotrienol (359.07 g/kg). Overall, there are significant differences in the content and bioaccessibility of total and individual tocols among commercial maize hybrids, allowing the selection of hybrids for animal production based not only on crude chemical composition but also on the content of phytochemicals.

A comparative study on in vitro and in vivo stomach-small intestinal and large intestinal digestion of plant protein meals in growing pigs.

This experiment evaluated the difference between computer-controlled simulated digestion and in vivo stomach-small intestinal or large intestinal digestion for growing pigs. Five diets including a corn-soybean meal basal diet and four experimental diets with rapeseed meal (RSM), cottonseed meal (CSM), sunflower meal (SFM), or peanut meal (PNM) were assigned to each group of five barrows installed terminal ileal cannula or distal cecal cannula in a 5 × 5 Latin square design. Ileal digesta and feces were collected for the determination of digestibility of dry matter (DM) and gross energy (GE) as well as digestible energy (DE) at terminal ileum and total tract. The large intestinal digestibility and DE were calculated by the difference between measurements obtained at the terminal ileum and those obtained from total tract. In vitro stomach-small intestinal digestibility and DE for diets and plant protein meals were determined by stomach-small intestinal digestion in a computer-controlled simulated digestion system (CCSDS). The in vitro large intestinal digestibility and DE of diets were determined in a CCSDS using ileal digesta and enzymes extracted from cecal digesta of pigs. The in vitro large intestinal digestibility and DE of four plant protein meals were determined by the difference between stomach-small intestinal and total tract digestion in the CCSDS. For the experimental diets, the in vitro ileal digestibility and DE were not different from corresponding in vivo values in basal diet and PNM diet, but greater than corresponding in vivo values for diets with RSM, CSM, and SFM (P < 0.05). No difference was observed between in vitro and in vivo large intestinal digestibility and DE in five diets. For the feed ingredients, the in vitro ileal digestibility and DE did not differ from corresponding in vivo ileal values in RSM and PNM but were greater than the in vivo ileal values in CSM and SFM (P < 0.05). The in vitro large intestinal GE digestibility and DE were not different from in vivo large intestinal values in RSM, CSM, and PNM, but lower than in vivo large intestinal values in SFM. This finding may relate to the higher fiber content of plant protein meals resulting in shorter digestion time of in vivo stomach-small intestine thus lower digestibility compared to in vitro, indicating it is necessary to optimize in vitro stomach-small intestinal digestion time.

Comparable in vitro and in vivo values are crucial to develop a novel in vitro digestion technique for growing pigs. The current study evaluated the difference between computer-controlled simulated digestion and in vivo stomach­small intestinal or large intestinal digestion for growing pigs. Five diets including a corn­soybean meal basal diet and four experimental diets with rapeseed meal (RSM), cottonseed meal (CSM), sunflower meal (SFM), or peanut meal (PNM) were used to compare the in vitro and in vivo digestion. Our study demonstrated that the in vitro ileal digestibility of energy was not different from corresponding in vivo values in basal diet and PNM diet, but greater than corresponding in vivo values for diets with RSM, CSM, and SFM. The in vitro stomach­small intestinal digestibility was greater than in vivo digestibility, resulting in less digestible substrates hydrolyzed by in vitro large intestinal fluid, whereas more digestible substrates can be digested by in vivo large intestine in plant protein meals. This difference may relate to the higher fiber content of plant protein meals resulting in shorter digestion time of in vivo stomach­small intestine thus lower digestibility compared to in vitro. Therefore, it is necessary to optimize in vitro stomach­small intestinal digestion time.

How Cooking Time Affects In Vitro Starch and Protein Digestibility of Whole Cooked Lentil Seeds versus Isolated Cotyledon Cells.

Lentils are sustainable sources of bioencapsulated macronutrients, meaning physical barriers hinder the permeation of digestive enzymes into cotyledon cells, slowing down macronutrient digestion. While lentils are typically consumed as cooked seeds, insights into the effect of cooking time on microstructural and related digestive properties are lacking. Therefore, the effect of cooking time (15, 30, or 60 min) on in vitro amylolysis and proteolysis kinetics of lentil seeds (CL) and an important microstructural fraction, i.e., cotyledon cells isolated thereof (ICC), were studied. For ICC, cooking time had no significant effect on amylolysis kinetics, while small but significant differences in proteolysis were observed (p < 0.05). In contrast, cooking time importantly affected the microstructure obtained upon the mechanical disintegration of whole lentils, resulting in significantly different digestion kinetics. Upon long cooking times (60 min), digestion kinetics approached those of ICC since mechanical disintegration yielded a high fraction of individual cotyledon cells (67 g/100 g dry matter). However, cooked lentils with a short cooking time (15 min) showed significantly slower amylolysis with a lower final extent (~30%), due to the presence of more cell clusters upon disintegration. In conclusion, cooking time can be used to obtain distinct microstructures and digestive functionalities with perspectives for household and industrial preparation.

Assessment of the Bioaccessibility of Elaeagnus angustifolia L. Flour and Its Use in Cracker Formulation.

In this study, in vitro digestion of polyphenols of oleaster (Elaeagnus angustifolia L.) flour (OF) and the potential use of OF in cracker production were studied. The results showed that 21.90 mg gallic acid equivalent (GAE)/g dry matter (DM) of total polyphenol (TP), 28.15 mg rutin equivalent (RE)/g DM of total flavonoid (TF) and 7709.13 mmol ascorbic acid equivalent (AAE)/100 g DM of antioxidant capacity (AC) were found in OF. The bioaccessibility and content of the OF polyphenols and their AC during in vitro digestion varied depending on the digestion stage. The stability of the OF polyphenols was higher in gastric stage (41.92%) than intestinal one (34.01%). OF was used to replace flour in the cracker formulation at the levels of 5, 10 and 15% (w/w) and increased the TP contents and AC of the crackers compared to the sample without OF (control). The addition of OF to the cracker dough increased the bioaccessibility of the cracker polyphenols. After digestion, bioaccessibilities of enriched crackers (107.90-126.81%) were found higher than that of control sample (93.73%). However, as the level of OF in cracker increased, bioaccessibility of polyphenols decreased. The results generally indicated that oleaster has a good potential for the enrichment of foods.

Effects of different proportions of stevia stalk on nutrient utilization and rumen fermentation in ruminal fluid derived from sheep.

Background: Stevia straw is a byproduct of sugar crop stevia. It is a good feed material because of richness in nutrients and active substances (steviosides and flavonoids). However, due to improper utilization such as piling, burning and so on, it became a large amount of wasted straw resources and lead to environmental pollution. Methods: We added 0%, 0.2%, 0.4%, 0.6%, 0.8%, 1.0% and 1.5% of stevia stalk to study the effects of different stevia stalk concentrations on nutrient utilization and rumen fermentation in sheep (based on sheep diet). In vitro fermentation method was used, with 17 repetitions for each treatment. All fermentation substrate based on sheep diet with different stevia stalk concentrations were fermented for 2 h, 6 h, 12 h, 24 h and 48 h, then the gas production, dry matter degradability (DMD), crude protein degradability (CPD), neutral detergent fiber degradability (NDFD), acid detergent fiber degradability (ADFD), pH, ammonia nitrogen (NH3-N) and volatile fatty acids (VFAs) were determined. Results: The results showed that at different fermentation time, the change trend of gas production in each teatment was basically same, but the maximum occurred in 1.0% treatment at 48 h. The DMD, CPD, NDFD and ADFD of sheep diets increased with fermentation time increasing, especially the CPD48h, NDFD48h and ADFD48h of diets in 0.8%, 1.0% and 1.5% treatments were significantly higher than those in control (P < 0.05). The pH of fermentation substrate in each treatment remained within the normal range of 6.21∼7.25. NH3-N24h-48hin 0.8%, 1.0% and 1.5% treatments were higher than that in control. At 6 h-12 h, the total acid content of 0.8% and 1.0% treatments were significantly higher than those of other treatments (P < 0.05), it reached the highest in 1.0% treatment. According to overall evaluation, effect ranking of stevia stalk on sheep nutrient utilization was as follows: 1.0% >0.8% >1.5% >0.4% >0.6% >0.2%. Overall, 1.0% stevia stalk could promote nutrient degradation and sheep rumen fermentation.

Characterization of Durum-Wheat Pasta Containing Resistant Starch from Debranched Waxy Rice Starch.

Durum wheat spaghetti samples prepared with increasing levels of resistant starch (RS) from debranched waxy rice starch (DWRS; i.e., 0, 5, 10, 15 g/100 g w/w) were analyzed for chemical composition, quality and sensory parameters and in vitro starch digestion. All the DWRS-containing spaghetti was "high in fibre", the dietary fiber content being > 6 g/100 g. In addition, spaghetti with the highest level of DWRS showed the highest RS content (p < 0.05), being 11.4 g/100 g dry matter. The starch hydrolysis index decreased (p < 0.05) as the level of DWRS increased, with a reduction of >20% comparing the 15-DWRS pasta to the control. DWRS had a negative impact on quality parameters, especially at higher DWRS levels. The use of DWRS shortened the optimal cooking time and impacted the samples' cooking loss, firmness, and stickiness. In addition, sensory analysis revealed differences among samples. However, irrespective of the level of DWRS in the recipe, the score for all attributes was > 5, which is considered the limit of acceptability. Substituting part of the semolina flour with DWRS increased the level of RS and the overall nutritional profile and affected the quality of semolina pasta, mainly at higher levels in the recipe.

Optimization of Compound Ratio of Exogenous Xylanase and Debranching Enzymes Supplemented in Corn-Based Broiler Diets Using In Vitro Simulated Gastrointestinal Digestion and Response Surface Methodology.

This experiment aimed to explore the zymogram of endo-xylanase (EX) and debranching enzymes (arabinofuranosidase [EA] and ferulic acid esterase [EF]) supplemented in the corn−soybean meal-based diet of broilers. An in vitro simulated gastrointestinal digestion model was adopted. According to single-factor, completely random design, the optimal supplemental levels of individual carbohydrase were determined by reducing sugars (RS) and in vitro dry matter digestibility (IVDMD). Response surface method (RSM) was used to predict the proper compound ratio of three carbohydrases. Results showed that shifts were different for feedstuffs such as corn−soybean meal−distillers dried grains with solubles, corn hull, and wheat bran, revealing that the net increase of RS or IVDMD distinctly dropped when degrading corn and related by-products by EX (p < 0.05). There was a significant quadratic relationship between the above response metrics and addition levels of each enzyme (p < 0.05). The determined dosage was 54 U/g EX, 5.0 U/g EA, and 0.4 U/g of EF, respectively. The optimistic zymogram of carbohydrases in corn basal substrates was judged by the IVDMD screening (R2 = 0.9089, p < 0.001). Conclusively, the in vitro assay and RSM were convenient and rapid methods for the optimization of xylan-degrading zymogram, and also testified asthenic hydrolysis of corn arabinoxylan by EX, thus highlighting the synergistic combinations with debranching enzymes.

A newly developed strain of Enterococcus faecium isolated from fresh dairy products to be used as a probiotic in lactating Holstein cows.

The objective of this study was to determine the ability of an isolated strain (EGY_NRC1) or commercial (NCIMB 11181) Enterococcus faecium as a probiotic for lactating cows. Two experiments were conducted: In Experiment 1, the effects of three levels (1, 2, and 3 g/kg diet, DM basis) of isolated and commercial E. faecium on in vitro ruminal fermentation kinetics, gas, methane (CH4) and nutrient degradability were determined. In Experiment 2, thirty multiparous Holstein cows (633 ± 25.4 kg body weight) with 7 days in milk, were randomly assigned to 3 treatments in a completely randomized design in a 60-day experiment. Cows were fed without any additives (control treatment) or supplemented with 2 g/kg feed daily of E. faecium EGY_NRC1 (contain 1.1 × 109 CFU/g) or commercial E. faecium NCIMB 11181 (contain 2 × 1012 CFU/g). Diets were prepared to meet cow's nutrient requirements according to NRC recommendations. Probiotic doses were based on the in vitro Experiment 1. Feed intake, digestibility, blood parameters and lactation performance were evaluated. In Experiment 1, the isolated E. faecium linearly and quadratically increased (P < 0.001) in vitro total gas production (TGP), the degradability of dry matter (dDM) and organic matter (dOM) while decreased (P < 0.05) methane (CH4) percent of TGP, NH3CH4 production, and pH. The commercial E. faecium increased TGP and decreased (P < 0.01) CH4 production, pH and increased the dDM and dOM, short chain fatty acids and ruminal NH3-N concentration. In Experiment 2, the isolated E. faecium increased (P < 0.01) total tract digestibility of DM, neutral and acid detergent fiber, daily milk production and feed efficiency compared to the control treatment without affecting feed intake and milk composition. Moreover, the isolated E. faecium increased (P < 0.05) the proportion of C18:1 trans-9, C18:2 cis-9-12 and C18:2 trans-10 cis-12. Both isolated and commercial E. faecium improved (P < 0.01) organic matter, crude protein and nonstructural carbohydrates digestibility, increased serum glucose (P = 0.002) and decreased serum cholesterol (P = 0.002). Additionally, both E. faecium strains decreased C23:0 (P = 0.005) in milk. In conclusion, the use of E. faecium (isolated and commercial) at 2 g/kg DM of feed improved feed efficiency and production performance, with superior effects on animal performance from isolated E. faecium compared to the commercial one.

Influence of Bile Salts and Pancreatin on Dog Food during Static In Vitro Simulation to Mimic In Vivo Digestion.

The addition of pancreatin and bile salts in different concentrations during in vitro digestion causes changes in the digestibility of crude protein (CP), fat, and dry matter (DM). The effects of bile salts and pancreatin on the digestibility of ether extract (EE), CP, and DM in developing a static in vitro digestion model for dogs were assessed using different concentrations of pancreatin (0, 1, 2.5, 5, and 10 g/L digestive solution) and bile salts (0, 2.5, 6.25, 12.5, and 25 g/L digestive solution). The data were analyzed using one-way analysis of variance. Digestibility of EE increased with the addition of bile salts (p < 0.05), whereas that of CP decreased with ≤0.25 g (1.0 g/L digestive solution) pancreatin. The digestibility of DM decreased significantly in all groups supplemented with ≥3.125 g (12.5 g/L digestive solution) bile salts and 0.25−2.5 g (1−10 g/L digestive solution) pancreatin and was the lowest with 6.25 g (25 g/L digestive solution) of bile salts (p < 0.05). These findings could facilitate the development of effective static in vitro digestion models for dogs.

Ensiling Grape Pomace With and Without Addition of a Lactiplantibacillus plantarum Strain: Effect on Polyphenols and Microbiological Characteristics, in vitro Nutrient Apparent Digestibility, and Gas Emission.

The present study investigated the effects of different grape pomace storage techniques on the effectiveness as feed on in vitro ruminant digestion efficiency. Grape pomace from an autochthonous red grape variety (cv Nero di Troia) was used as fresh (GP) or ensiled, both without additives (SIL) and with the addition of a bacterial strain, Lactiplantibacillus plantarum 5BG (SIL+). All the different storage treatments were subject to chemical and microbiological evaluation, as well as in vitro digestibility, and gas production. Microbiological data revealed the good quality of grape pomace and silages due to the lactic acid bacteria populations and low presence, or absence, of undesirable microorganisms. The addition of L. plantarum 5BG influenced the chemical characteristics of the silage (SIL+). Ensiling technique deeply changed the polyphenolic composition, reducing anthocyanins, flavonols, and flavanols (condensed tannins precursors), particularly when L. plantarum 5BG was added. Antioxidant capacity was reduced by ensiling, in correlation with the polyphenolic content decrease. The oxygen radical absorbance capacity (ORAC) value of SIL+ was the lowest (P < 0.01) and its total phenol content was lower than SIL (P < 0.01). No statistical differences were observed between GP, SIL, and SIL+ on the antioxidant capacity by TEAC assay (P > 0.05). Ensiling did not affect the grape pomace nutrient profile, except for the reduction in NFC content. Apparent in vitro digestibility showed how ensiling increased dry matter (DM), organic matter (OM), neutral detergent fiber (NDF), crude protein (CP), ether extract (EE), and non-fiber carbohydrates (NFC) disappearance (P < 0.01), particularly with the L. plantarum 5BG inoculation. Moreover, SIL+ showed the lowest propionic acid (P < 0.05) and the highest methane (P < 0.01), butyric acid (P < 0.01), and nitrogen (P < 0.05) in vitro production. Ensiling GP resulted in a better in vitro digestibility, particularly if L. plantarum 5BG strain is added, probably due to the reduction of flavanols and their lower microbial activity inhibition.

Digestion of bamboo compared to grass and lucerne in a small hindgut fermenting herbivore, the guinea pig (Cavia porcellus).

Bamboo is an enigmatic forage, representing a niche food for pandas and bamboo lemurs. Bamboo might not represent a suitable forage for herbivores relying on fermentative digestion, potentially due to its low fermentability. To test this hypothesis, guinea pigs (n = 36) were used as model species and fed ad libitum with one of three forages (bamboo, lucerne, or timothy grass) in a fresh or dried state, with six individuals per group, for 3 weeks. The nutrient composition and in vitro fermentation profile of bamboo displayed low fermentation potential, i.e. high lignin and silica levels together with a gas production (Hohenheim gas test) at 12 h of only 36% of that of lucerne and grass. Although silica levels were more abundant in the leftovers of (almost) all groups, guinea pigs did not select against lignin on bamboo. Dry matter (DM) intake was highest and DM digestibility lowest on the bamboo forage. Total short-chain fatty acid levels in caecal content were highest for lucerne and lowest for grass and bamboo. Bamboo-fed guinea pigs had a lower body weight gain than the grass and lucerne group. The forage hydration state did not substantially affect digestion, but dry forage led to a numerically higher total wet gut fill. Although guinea pigs increased DM intake on the bamboo diet, the negative effects on fermentation of lignin and silica in bamboo seemed overriding. For herbivores that did not evolutionary adapt, bamboo as an exclusive food resource can be considered as inadequate.

Degradability, in vitro fermentation parameters, and kinetic degradation of diets with increasing levels of forage and chitosan.

Chitosan is the second most important natural biopolymer in the world, extracted from crustaceans, shrimps, and crabs and can modulate rumen fermentation. Our hypothesis is that the addition of chitosan alters the fermentation patterns of different diets for ruminants. This study aimed to evaluate the effects of different levels of chitosan and forage on in vitro dry degradation kinetics and fermentation in a gas production system. The chitosan levels (0, 1625, 3,500, or 7,500 mg/kg of dry matter [DM]) were arranged in a completely randomized block design, and for in vitro ruminal fermentation assay, we used a split splot arrangement. Into the incubator, all chitosan levels were distributed in the four jars, and the forage levels varying on 100, 65, 50, 35, and 20 on DM basis. There was an interaction effect for chitosan and forage levels (P ≤ 0.05) on IVDMD; IVOMD. IVDCP and IVDNDF. Chitosan negatively affected IVDMD in all roughage levels evaluated. The pH and ammonia concentration present effect only for roughage levels and incubation hours. The chitosan did not change (P = 0.3631) the total short-chain fatty acid concentration (overall mean = 21.19 mmol/L) and the C2:C3 ratio (overall mean = 5.85). The IVDCP showed the same decreasing quadratic behavior (P < 0.0001). The increasing chitosan addition increases (P < 0.0001) the gas production and decreases (P < 0.0001) the lag time (parameter C) of diets with greater concentrate participation, characterizing greater efficiency in the degradability of the diet, confirming its potential use in diets for ruminants. Chitosan changes in vitro dry degradation kinetics and fermentation at the minimum dose of 1,722 mg/kg DM for all diets. The roughage level influenced the in vitro nutrients degradability and cumulative gas production.

Effect of Donor Diet on In Vitro Digestibility of Forages by Fecal Inoculate.

Previous research indicates equine fecal inoculates produce comparable results to cecal fluid when used for in vitro procedures to analyze dry matter digestibility (DMD). Equine hindgut microbial communities represented in fecal samples have been shown to be affected by diet. The study's objective was to determine the effect of the donor diet on in vitro DMD when fecal donors were fed high starch, or high fiber diets. Six Quarter Horses were used in a crossover design to compare the effects of a grain versus forage diet on in vitro digestion of forages ranging from: CP 7.7 to 16.4 %DM, NDF 53 to 72 %DM. Feces from each horse were obtained on day 22 of each period and used to inoculate in vitro fermentation vessels in order to evaluate the effect of donor diet on the DMD, neutral detergent fiber digestibility (NDFD), and acid detergent fiber digestibility (ADFD) of four forages. Data were analyzed using the MIXED procedure of SAS to evaluate digestibility differences in the diet by forage composition interaction. Fecal samples from horses on the grain diet promoted higher NDFD of forages with high NDF and low CP when CP and NDF were used as covariates (P = .04 and .03). There was a horse effect on DMD and NDFD (P ≤ .05). Findings suggest diet may influence the hindgut microbiome's ability to digest neutral detergent fiber and should be considered when selecting equine fecal samples for in vitro digestion procedures.