Comparative in vitro fermentation activity in the canine distal gastrointestinal tract and fermentation kinetics of fiber sources

The current study aimed to evaluate the variation in fermentation activity along the distal canine gastrointestinal tract (GIT, Exp. 1). It also aimed to assess fermentation kinetics and end product profiles of 16 dietary fibers for dog foods using canine fecal inoculum (Exp. 2). For Exp. 1, digesta were collected from the distal ileum, proximal colon, transverse colon, and rectum of 3 adult dogs. Digesta per part of the GIT were pooled for 3 dogs, diluted (1:25, wt/vol), mixed, and filtered for the preparation of inoculum. A fructan, ground soy hulls, and native potato starch were used as substrates and incubated for cumulative gas production measurement as an indicator of the kinetics of fermentation. In addition, fermentation bottles with similar contents were incubated but were allowed to release their gas throughout incubation. Fermentation fluid was sampled at 4, 8, 12, 24, 48, and 72 h after initiation of incubation, and short-chain fatty acids and ammonia were measured. Results showed comparable maximal fermentation rates for rectal and proximal colonic inocula (P > 0.05). Production of short-chain fatty acids was least for the ileal and greatest for the rectal inoculum (P < 0.05). Therefore, for in vitro studies, fecal microbiota can be used as an inoculum source but may slightly overestimate in vivo fermentation. Experiment 2 evaluated the gas production, fermentation kinetics, and end product profiles at 8 and 72 h of incubation for citrus pectin, 3 fructans, gum arabic, 3 guar gums, pea fiber, peanut hulls, soy fiber, sugar beet fiber, sugar beet pectin, sugar beet pulp, wheat fiber, and wheat middlings. Feces of 4 adult dogs were used as an inoculum source. Similar techniques were used as in Exp. 1 except for the dilution factor used (1:10, wt/vol). Among substrates, large variations in fermentation kinetics and end product profiles were noted. Sugar beet pectin, the fructans, and the gums were rapidly fermentable, indicated by a greater maximal rate of gas production (R(max)) compared with all other substrates (P < 0.05), whereas peanut hulls and wheat fiber were poorly fermentable, indicated by the least amount of gas produced (P < 0.05). Sugar beet fiber, sugar beet pulp, soy fiber, and wheat middlings were moderately fermentable with a low R(max). Citrus pectin and pea fiber showed a similar low R(max), but time at which this occurred was later compared with sugar beet fiber, sugar beet pulp, soy fiber, and wheat middlings (P < 0.05). Results of this study can be used to formulate canine diets that stimulate dietary fiber fermentation along the distal GIT that may optimize GIT health and stimulate the level of satiety in dogs.     

Adaptation of healthy adult cats to select dietary fibers in vivo affects gas and short-chain fatty acid production from fiber fermentation in vitro

Nine young adult (1.73 ± 0.03 yr) male cats were used to determine the effects of microbial adaptation to select dietary fiber sources on changes in pH in vitro and on total and hydrogen gas, short-chain fatty acid (SCFA), and branched-chain fatty acid (BCFA) production. Cats were adapted to diets containing 4% cellulose, fructooligosaccharides (FOS), or pectin for 30 d before fecal sampling. Each cat was used as a single donor, and fecal inoculum was reacted with each of the aforementioned fiber substrates. Adaptation to dietary FOS resulted in a greater change in pH when exposed to FOS than pectin (adaptation × substrate, P < 0.001). When exposed to the FOS substrate, adaptation to dietary FOS or pectin increased hydrogen gas production (adaptation × substrate, P = 0.021). Adaptation to dietary FOS increased acetate and total SCFA production when exposed to FOS substrate in vitro (adaptation × substrate, P = 0.001). When exposed to the FOS substrate, propionate production tended to increase with adaptation to dietary cellulose (adaptation × substrate, P = 0.060). The BCFA + valerate tended to decrease with adaptation to dietary FOS when exposed to FOS substrate in vitro (adaptation × substrate, P = 0.092). Fructooligosaccharides resulted in the greatest change in pH and production of total gas (P < 0.001), hydrogen gas (P < 0.001), acetate (P < 0.001), propionate (P < 0.001), butyrate (P < 0.001), total SCFA (P < 0.001), and total BCFA + valerate production (P < 0.001). Adaptation to the FOS or pectin diet increased production of hydrogen gas with FOS and pectin substrates. Adaptation to pectin increased (P = 0.033) total gas production with FOS and pectin substrates. Overall, adaptation to either FOS or pectin led to greater SCFA and gas production, but adaptation to FOS resulted in the greatest effect overall.     

Evaluation of fermentable oligosaccharides in diets fed to dogs in comparison to fiber standards

Blends of fermentable oligosaccharides in combination with nonfermentable fiber, cellulose, were evaluated for their ability to serve as dietary fibers in dog foods. Using a 6 x 6 Latin square design, 6 diets were evaluated that contained either no supplemental fiber, beet pulp, cellulose, or blends of cellulose, fructooligosaccharides, and yeast cell wall added at 2.5% of the diet. Six ileal-cannulated dogs were fed 175 g of their assigned diet twice daily. Chromic oxide served as a digestibility marker. Nutrient digestibility, fecal microbial populations, fermentative end products, and immunological indices were measured. Total tract DM and OM digestibilities were lowest (P < 0.05) for the cellulose treatment. Crude protein digestibility was lower (P < 0.05) for the treatments containing carbohydrate blends. The cellulose treatment had the lowest (P < 0.05) concentration of bacteria, and all diets containing fermentable fiber had greater (P < 0.05) fecal bifidobacteria concentrations compared with the diets without supplemental fermentable fiber. Lactobacilli concentrations tended to be greater (P < 0.08) in treatments containing fermentable fiber compared with the cellulose treatment. Bifidobacteria and lactobacilli concentrations were similar for the beet pulp treatment compared with the fermentable oligosaccharide blends. Total fecal short-chain fatty acid concentration was greater for the beet pulp treatment (P < 0.05) compared with the control and cellulose treatments. The treatments containing fermentable fiber had greater (P < 0.05) fecal butyrate concentrations compared with cellulose and control treatments. Immune indices were not affected by treatment. Our results suggest that dog foods containing blends of fermentable and nonfermentable carbohydrates produce similar physiological results as dog food containing beet pulp as a fiber source. Therefore, blends of these carbohydrates could be useful substitutes for beet pulp in dog foods.     

In vitro fermentation characteristics of selected oligosaccharides by swine fecal microflora

The objective of this study was to quantify the fermentation characteristics of oligosaccharides present in feed ingredients or isolated for dietary supplementation. Substrates studied included short-chain fructooligosaccharides, medium-chain fructooligosaccharides, long-chain fructooligosaccharides, raffinose, stachyose, soy solubles, granular and liquid forms of transgalactooligosaccharides, glucooligosaccharides, mannanoligosaccharides, and xylooligosaccharides. Three healthy pigs that had never received antibiotics served as sources of fecal inoculum. Each substrate was fermented in vitro; samples were taken at 0, 2, 4, 8, and 12 h, and pH change and short-chain fatty acid (SCFA) and gas production determined. Gas production at 12 h did not differ (P > 0.05) among all fructooligosaccharides, transgalactooligosaccharides, soy solubles, and xylooligosaccharides. Raffinose, stachyose, and raffinose + stachyose fermentation resulted in the greatest (P < 0.05) gas production at 12 h of all substrates tested. The rate of gas production was greatest (P < 0.05) for stachyose and least (P < 0.05) for glucooligosaccharides and mannanoligosaccharides. Substrate did not affect (P > 0.05) time to attain maximal rate of gas production. The pH at 12 h for all fructooligosaccharides and xylooligosaccharides did not differ (P > 0.05). The pH values at 12 h for raffinose, stachyose, and raffinose + stachyose were highest (P < 0.05) compared with all other substrates. Total SCFA production at 12 h was similar for all fructooligosaccharides and transgalactooligosaccharides, glucooligosaccharides, and soy solubles. Total SCFA production was greatest (P < 0.05) for xylooligosaccharides, stachyose, and raffinose + stachyose, and least (P < 0.05) for mannanoligosaccharides and raffinose. Stachyose fermentation resulted in the greatest (P < 0.05) rate and earliest time to attain maximal rate of SCFA production. All oligosaccharides studied were readily fermentable but varied in amount and type of SCFA produced. Fermentation of the pure forms of oligosaccharides contained in soy solubles resulted in greater gas production and higher pH compared with soy solubles. The oligosaccharides in the soy solubles matrix seemed to behave differently than their pure counterparts. The high rates of fermentation of most oligosaccharides tested indicate that they may serve as fermentable carbohydrate sources in the terminal small intestine or large intestine of swine.     

肉仔鸡盲肠微生物体外发酵非消化寡糖的比较研究

为了评价棉子糖、水苏糖、果寡糖、甘露寡糖等4种非消化寡糖的发酵特性,以采食无抗生素日粮的肉仔鸡盲肠微生物为菌源,通过体外发酵试验比较短链脂肪酸(SCFA)的产量及产生速率。结果表明:棉子糖发酵6 h后,SCFA产量即迅速增加,显著高于其他发酵底物(P<0.05),而且在各发酵底物产生SCFA中乙酸比例最大、丁酸比例最小(P<0.05);24 h,SCFA产量达到最大,丁酸比例在各发酵组中也增至最大。水苏糖和果寡糖的发酵速率紧随棉子糖之后,但6 h和12 h的SCFA产量仅为棉子糖的一半左右,24 h的SCFA总产量分别达到棉子糖的83.81%和75.84%。甘露寡糖的发酵速率最慢,6、12、24 h的SCFA总产量仅为棉子糖的23.81%、18.90%、40.94%。     

Use of the in vitro cumulative gas production technique for pigs: an examination of alterations in fermentation products and substrate losses at various time points

An experiment was conducted to examine changes in VFA and ammonia concentrations at different time points using 4 fermentable carbohydrate-rich feed ingredients as substrates and feces of unweaned piglets as inoculum. Fecal inoculum was collected, pooled, and mixed from 9 specially raised (no creep feed or antibiotics) crossbred piglets at 3 wk of age. Inulin, lactulose, molasses-free sugar beet pulp, and wheat starch were used as substrates and were fermented in vitro for 72 h (3 replicates per substrate). Cumulative gas production was measured as an indicator of the kinetics of fermentation. In addition, 3 bottles of substrate per time point with similar contents (amounts of substrate, inoculum, and media) were incubated but were allowed to release their gas throughout incubation. For these latter bottles, fermentation fluid was sampled at incubation time points including every hour between 1 and 24 h and at 48 h, and fermentation end products (VFA, lactate, and ammonia) and OM disappearance were measured. Dry matter and ash were analyzed from the postfermentative samples. The pH of the contents from these bottles was also recorded. The correlation in time between fermentation end products and cumulative gas produced was determined. The results showed that the prolongation of fermentation to 72 h, especially in the case of fast-fermenting inulin and lactulose, may lead to a different end product profile (P < 0.001) compared with the profile observed at the time at which most of the substrate has disappeared. Therefore, we concluded that the fermentation product profile at the end of in vitro fermentation at a specific time point cannot be used to compare fermentability of carbohydrate sources with different fermentation kinetics in terms of gas production.     

Fermentation of animal components in strict carnivores: a comparative study with cheetah fecal inoculum

The natural diet of felids contains highly digestible animal tissues but also fractions resistant to small intestinal digestion, which enter the large intestine where they may be fermented by the resident microbial population. Little information exists on the microbial degradability of animal tissues in the large intestine of felids consuming a natural diet. This study aimed to rank animal substrates in their microbial degradability by means of an in vitro study using captive cheetahs fed a strict carnivorous diet as fecal donors. Fresh cheetah fecal samples were collected, pooled, and incubated with various raw animal substrates (chicken cartilage, collagen, glucosamine-chondroitin, glucosamine, rabbit bone, rabbit hair, and rabbit skin; 4 replicates per substrate) for cumulative gas production measurement in a batch culture technique. Negative (cellulose) and positive (casein and fructo-oligosaccharides; FOS) controls were incorporated in the study. Additionally, after 72 h of incubation, short-chain fatty acids (SCFA), including branched-chain fatty acids (BCFA), and ammonia concentrations were determined for each substrate. Glucosamine and glucosamine-chondroitin yielded the greatest organic matter cumulative gas volume (OMCV) among animal substrates (P < 0.05), whereas total SCFA production was greatest for collagen (P < 0.05). Collagen induced an acetate production comparable with FOS and a markedly high acetate-to-propionate ratio (8.41:1) compared with all other substrates (1.67:1 to 2.97:1). Chicken cartilage was rapidly fermentable, indicated by a greater maximal rate of gas production (R(max)) compared with all other substrates (P < 0.05). In general, animal substrates showed an earlier occurrence for maximal gas production rate compared with FOS. Rabbit hair, skin, and bone were poorly fermentable substrates, indicated by the least amount of OMCV and total SCFA among animal substrates (P < 0.05). The greatest amount of ammonia production among animal substrates was measured after incubation of collagen and rabbit bone (P < 0.05). This study provides the first insight into the potential of animal tissues to influence large intestinal fermentation in a strict carnivore, and indicates that animal tissues have potentially similar functions as soluble or insoluble plant fibers in vitro. Further research is warranted to assess the impact of fermentation of each type of animal tissue on gastro-intestinal function and health in the cheetah and other felid species.     

In vitro fermentation of various fiber and starch sources by pig fecal inocula

Freeze-dried ileal effluent (1% wt/vol) from cannulated pigs fed rice-based diets with the inclusion of either animal protein (CON), animal protein plus potato starch (PS), animal protein plus sugar beet pulp (SBP), or animal protein plus wheat bran (WB) was incubated anaerobically at pH 6.0 in fermenters containing 5% (wt/vol) fecal slurry comprising mineral salts medium and 50 g/L of fresh feces from pigs fed the same diets as the cannulated pigs. Samples were collected from the fermenters at 0, 2, 4, 12, 24, and 48 h during in vitro fermentation for measuring nonstarch polysaccharides (NSP), starch, and short-chain fatty acids (SCFA). Results showed that the major SCFA produced were acetate, propionate, and butyrate. The inclusion of soluble dietary fiber (diet SBP) caused the highest concentrations of acetate, propionate, butyrate, and total SCFA, whereas the increase in the production of propionate resulting from the addition of insoluble dietary fiber (diet WB) only occurred at the initial stages during 48 h in vitro fermentation. At all sampling occasions (except for 4 h), the levels of butyrate were increased (P < 0.01) by resistant starch compared with fiber sources, showing that a higher level of butyrate can be achieved through microbial fermentation by potato starch. Lowered (P < 0.05) butyrate concentrations were observed with diet WB during in vitro fermentation. With the inclusion of fiber sources, the energy originating from SCFA was similar to that from NSP disappearance, whereas the values were lower (P < 0.05) from NSP disappearance than for SCFA generated without fiber sources supplemented. We conclude that more substrate is available in ileal effluent with the addition of soluble dietary fiber, and an increased level of butyrate could be achieved through microbial fermentation by resistant starch.     

Fruit and vegetable fiber fermentation by gut microflora from canines

The objective of this study was to assess fermentability by canine gut microflora to include shortchain fatty acid (SCFA) production, organic matter (OM) disappearance, and gas production of vegetable and fruit fiber sources compared to fiber standards (psyllium, citrus pectin, and Solka Floc). Fiber sources included apple pomace, carrot pomace, flaxseed, fruit blend (mixture of peach, almond, nectarine, and plum), grape pomace, pea hulls, pistachio, and tomato pomace. Substrates were fermented in vitro for 4, 12, and 24 h with fecal flora obtained from three healthy dogs. Citrus pectin had the highest OM disappearance, SCFA production, and gas production at all times of fermentation; psyllium was intermediate and Solka Floc was lowest. A wide variation in fermentability was noted among the vegetable and fruit fiber sources. Apple pomace, carrot pomace, and flaxseed had the greatest fermentability as assessed by OM disappearance. Pea hulls and tomato pomace had intermediate OM disappearances, and fruit blend, grape pomace, and pistachio were poorly fermented. Carrot pomace produced the largest amounts of gas and SCFA. Apple pomace produced high concentrations of gas but intermediate concentrations of SCFA. Pea hulls and tomato pomace produced intermediate concentrations of gas and SCFA, whereas flaxseed, fruit blend, grape pomace, and pistachio produced low amounts of these fermentation products. For all substrates collectively, OM disappearance was highly correlated with both gas production (r2 = 0.782 and 0.723 for 12- and 24-h values, respectively) and SCFA production (r2 = 0.737 and 0.738 for 12- and 24-h values, respectively). In general, OM disappearance, gas production, and SCFA production were related to the insoluble:soluble fiber ratio in the samples; as the insoluble:soluble ratio decreased (increased soluble fiber), the OM disappearance, gas production, and SCFA production increased.     

In vitro fermentation characteristics of different carbohydrate sources in two dog breeds (German shepherd and Neapolitan mastiff)

Few studies have been published on the normal intestinal biota of canines unlike the wealth of information regarding livestock animal species. The in vitro gas production technique (IVGPT) including measurements of accumulating gas during fermentation and end‐product determinations allows obtaining a complete picture of microbial activity kinetics. The aim of this study was to study the in vitro fermentation characteristics of different carbohydrate sources using inocula from two dog breeds (German Shepherd and Neapolitan mastiff). Faeces sampled from rectum of two GS and NM adult dogs, fed the same dry food, were used as inocula. The samples, diluted and filtered, were incubated at 39 °C under anaerobic condition with nine substrates different for carbohydrate composition (rice, corn, potato, spelt, pure cellulose, beet pulp, wheat bran, inulin and fructo‐oligosaccharide). Gas production was recorded 17 times using a manual pressure transducer. After 48 h, the fermentation was stopped and fermenting liquor was analysed for pH and volatile fatty acids (VFA). Organic matter digestibility (OMD) was calculated as difference after burning the residuals. OMD, gas production and end‐products were significantly correlated with chemical composition of substrates, in particular carbohydrate fractions (total dietary fibre and starch), confirming the effectiveness of the IVGPT in evaluating dog feeds. Concerning the comparison between breeds significant differences (p < 0.01) were found for OMD, gas production, fermentation kinetic parameters and end‐products, suggesting a different pathway of fermentation and consequently, a different anaerobic population.     

梅山与长白母猪粪样微生物体外发酵八种纤维底物的特性比较

本研究旨在比较不同品种母猪(梅山猪与长白猪)粪样中微生物对不同纤维底物的体外降解能力,同时分析粪样中的主要纤维降解菌数量。采集梅山(n=5)和长白(n=5)母猪新鲜粪样作为发酵接种物,以果胶、纤维素、菊粉、麦壳、麸皮、木聚糖、玉米芯渣及苜蓿作为纤维底物进行体外发酵,测定产气量和发酵液挥发性脂肪酸(volatile fatty acid,VFA)浓度。粪样同时用于提取细菌总核酸,变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)和real-time PCR对菌群区系进行分析。体外发酵结果表明,从发酵后9 h至96 h结束,长白猪接种物各组的累积产气量、有机物校正产气量分别显著(P<0.05)和极显著(P<0.01)高于梅山猪,长白猪T_1/2和T_max显著低于梅山猪(P<0.05),且发酵终产物中乙酸和总VFA浓度显著高于梅山猪接种物(P<0.05),整个发酵过程中不同底物间累积产气量差异显著(P<0.05),累积产气量从高到低的组别依次为:菊粉>麸皮>果胶>苜蓿>麦壳>玉米芯渣>木聚糖>纤维素。DGGE分析表明,梅山与长白母猪粪样菌群图谱中存在许多共同条带。Real-time PCR定量分析表明,梅山母猪粪样中的总细菌的16S rRNA基因拷贝数显著高于长白母猪(P<0.05),而拟杆菌、产琥珀酸丝状杆菌、黄化瘤胃球菌和白化瘤胃球菌等纤维降解菌数量及其占总菌比例差异均不显著(P>0.05)。结果显示,尽管2个品种母猪粪样中纤维降解菌数量无显著差异,但是长白母猪粪样微生物体外发酵纤维底物的能力高于梅山母猪。     

In Vitro Gas Production from Batch Cultures of Stomach and Hindgut Digesta of Horses Adapted to a Prebiotic Dose of Fructooligosaccharides and Inulin

Fructooligosaccharides (FOS) and inulin may modulate hindgut fermentation. It was tested if digesta batch cultures taken from horses adapted to FOS and inulin show different fermentation compared with such taken from nonsupplemented horses. Six horses received 0.15 g FOS and inulin/kg body weight/d via Jerusalem artichoke meal (JAM) upon a hay-based diet; six horses received corncob meal without grains (CMG) as placebo. The horses were euthanized after 20 days. Digesta samples were taken from stomach, cecum, ventral colon ascendens (VCA), and colon transversum (CT). Digesta batch cultures were incubated 48 hours to measure in vitro gas production as well as pre- and post-incubation pH and oxidation-reduction potential (ORP). A distinct fermentation of the surplus of fructans present in the inoculum was found with JAM-adapted batch cultures. Gas production was accelerated in inoculated gastric contents of horses adapted to JAM compared with CMG adapted ones (7.8 vs. 16.4 hours to achieve half of the 48 hours gas quantity, respectively; P > .05). Although buffered, pH decreased during fermentation. Postincubation pH was lower with JAM than CMG-adapted batch cultures (P > .05). Preinoculation ORP was lower with stomach batch cultures adapted to CMG than with such adapted to JAM. The ORP increased twofold from pre- to post-incubation with the latter. Asymptotic maximal gas production decreased gradually using cecum, VCA, or CT digesta. Parts of FOS and inulin of digesta are fermented in the stomach, which reduce possible effects on hindgut fermentation. Elevated fermentation may considerably impact stomach health.     

Fiber fermentability effects on energy and macronutrient digestibility, fecal traits, postprandial metabolite responses, and colon histology of overweight cats

Considering the different potential benefits of divergent fiber ingredients, the effect of 3 fiber sources on energy and macronutrient digestibility, fermentation product formation, postprandial metabolite responses, and colon histology of overweight cats (Felis catus) fed kibble diets was compared. Twenty-four healthy adult cats were assigned in a complete randomized block design to 2 groups of 12 animals, and 3 animals from each group were fed 1 of 4 of the following kibble diets: control (CO; 11.5% dietary fiber), beet pulp (BP; 26% dietary fiber), wheat bran (WB; 24% dietary fiber), and sugarcane fiber (SF; 28% dietary fiber). Digestibility was measured by the total collection of feces. After 16 d of diet adaptation and an overnight period without food, blood glucose, cholesterol, and triglyceride postprandial responses were evaluated for 16 h after continued exposure to food. On d 20, colon biopsies of the cats were collected under general anesthesia. Fiber addition reduced food energy and nutrient digestibility. Of all the fiber sources, SF had the least dietary fiber digestibility (P < 0.05), causing the largest reduction of dietary energy digestibility (P < 0.05). The greater fermentability of BP resulted in reduced fecal DM and pH, greater fecal production [g/(cat × d); as-is], and greater fecal concentration of acetate, propionate, and lactate (P < 0.05). For most fecal variables, WB was intermediate between BP and SF, and SF was similar to the control diet except for an increased fecal DM and firmer feces production for the SF diet (P < 0.05). Postprandial evaluations indicated reduced mean glucose concentration and area under the glucose curve in cats fed the SF diet (P < 0.05). Colon mucosa thickness, crypt area, lamina propria area, goblet cell area, crypt mean size, and crypt in bifurcation did not vary among the diets. According to the fiber solubility and fermentation rates, fiber sources can induce different physiological responses in cats, reduce energy digestibility, and favor glucose metabolism (SF), or improve gut health (BP).     

Absorption of carbohydrate-derived nutrients in sows as influenced by types and contents of dietary fiber

The current investigation was undertaken to study the absorption and plasma concentration of carbohydrate-derived nutrients [glucose, short-chain fatty acids (SCFA), and lactate] and the apparent insulin production in sows fed diets containing contrasting types and contents of dietary fiber. Six sows were fed 3 experimental diets, low fiber (LF; 177 g of dietary fiber and 44 g of soluble fiber/kg of DM), high soluble fiber (HF-S; 429 g of dietary fiber and 111 g of soluble fiber/kg of DM), and high insoluble fiber (HF-I; 455 g of dietary fiber and 74 g of soluble fiber/kg of DM), in a repeated crossover design. Variations in dietary concentration and solubility of dietary fiber were obtained by substituting starch-rich wheat and barley in the LF diet with dietary fiber-rich co-products (sugar beet pulp, potato pulp, pectin residue, brewers spent grain, pea hulls, and seed residue, which have distinct physicochemical properties). The main carbohydrate component of the LF diet was starch and nonstarch polysaccharides (cellulose and noncellulosic polysaccharides) for the 2 high dietary fiber diets. Consumption of the LF diet resulted in increased and rapid glucose absorption at 0 to 4 h postfeeding. With the HF-I diet, the glucose absorption pattern was similar but at a decreased rate, whereas it was decreased and delayed with the HF-S diet (diet, P < 0.001; time, P < 0.001). These differences were also reflected in the insulin response. The quantitative absorption of SCFA at 0 to 10 h postfeeding was greater when feeding the HF-S diet compared with the LF diet (P < 0.001) and intermediate when feeding the HF-I diet (P < 0.001). The study showed that feeding the high dietary fiber diets resulted in a increased and more uniform uptake of SCFA than when feeding the LF control. Moreover, the HF-S diet reduced diurnal variation in glucose and insulin concentrations.     

Use of faeces as an alternative inoculum to caecal content to study in vitro feed digestibility in domesticated ostriches (Struthio camelus var. domesticus)

1. In order to find an alternative source of inoculum to caecal content for studying the in vitro feed digestibility in domesticated ostriches (Struthio camelus var. domesticus), caecal content and faeces of 4 male birds were used as inocula for an in vitro gas production trial. 2. About 1 g of each of 5 substrates (maize silage, CS; alfalfa hay, AH; barley, BG; soybean meal, SM; beet pulp, BP) was weighed, in quadruplicate per inoculum, in 120 ml flasks; 75 ml of anaerobic medium and 4 ml of reducing solution were added and flasks were kept at 39 degrees C. Caecal content and faeces were diluted respectively 1 : 2 (CI) and 1 : 4 (FI) with an anaerobic medium and were injected into the respective flasks (10 ml). 3. Gas production was recorded 22 times up to 120 h of incubation and fermentation characteristics (for instance, degraded organic matter, OMd; potential gas production, A; maximum fermentation rate, Rmax; time at which it is reached, Tmax; pH; volatile fatty acid, VFA; ammonia) were studied for each inoculum and substrate. 4. CI and FI showed significant differences in Tmax (16.37 vs 18.47 h, respectively), propionic (16.47 vs 12.07 mmoles/l) and butyric acid (6.50 vs 7.98 mmoles/l) and ammonia concentration (17.18 vs 19.95 mmoles/l). The substrates, according to their chemical composition, showed different fermentation characteristics. However, the regression equations able to estimate some fermentation characteristics of the caecum from those of faeces were statistically significant and showed R2-values ranging from 0.87 to 0.99. 5. The differences in fermentation pathways of the two inocula did not appear to influence the rate and extent of OM digestion. Faecal fermentation predicted rates and extent of OM digestion by caecal fermentation in ostriches; consequently, the faeces could be considered as an alternative to caecal content to study feed digestibility in the species, although there is a need to undertake further research.     

Rumen fermentation and degradability in buffalo and cattle using the in vitro gas production technique

An in vitro trial was conducted to investigate the effect of different inoculum sources (buffalo vs. cattle) on rumen fermentation and degradability. Incubations were carried out using rumen fluid obtained from buffalo or cattle fed the same diet [60% grass hay and 40% concentrate; 18 kg dry matter (DM)/day]. The fermentation kinetics of eight feeds commonly used in ruminant nutrition (alfalfa hay, barley meal, beet pulp, corn meal and silage, ryegrass hay and silage and soya bean meal s.e.) were studied with the in vitro gas production technique and rumen fermentation parameters (substrate disappearance, pH and volatile fatty acids production) were determined after 120 h of incubation. The linear relationship indicates that the microbial metabolic pathways of the two inocula for all the substrates were qualitatively similar, albeit often quantitatively different. In this in vitro study, a significant influence of rumen inoculum (buffalo vs. cow) on fermentation and degradability of the examined substrates was found. The differences in buffalo and cattle rumen fermentation can be explained with a different microbial activity of the two ruminant species, because of different amount of microbial population or microbial population constituted by different species of bacteria and protozoa.     

Effects of Moringa oleifera seed extract on rumen fermentation in vitro.

Moringa oleifera is a pantropical tree of the family Moringaceae. A previously undescribed property of an aqueous extract from the seeds of this plant is the modulation of ruminal fermentation patterns, especially protein degradation, as demonstrated in a short-term batch incubation system. Gas, short chain fatty acids (SCFA) and cellulolytic enzyme activities were determined as general fermentation parameters. A dot blot assay able to directly detect true protein in rumen fluid samples was used to quantify protein degradation. For complex substrates the interpretation of protein degradation profiles was amended by polyacrylamide gel electrophoresis (PAGE) of the samples. When incubated with pure carbohydrates at a concentration of 1 mg ml(-1), the extract reduced microbial degradation of the model protein, bovine serum albumin (BSA), such that its concentration was at least 40% above the control after 12 h of incubation. Total protein degradation was thus delayed by approximately 9 h. When fermented along with wheat straw, leaf protein (Rubisco) was almost entirely protected during 12 h of fermentation. The degradation of soy proteins was retarded by at least 4-6 h, depending on the protein band. There were strong side effects on the fermentation of pure cellulose (SCFA yield-60% after 12 h), whereas cellobiose and starch fermentation were less affected (-18 and -8%, respectively). When the complex substrates were fermented, SCFA yield was reduced by approximately 30% after 12 h. In our work we clearly demonstrate the efficacy of the new substance, which is neither a tannin nor a saponin, in an in vitro system, using pure as well as complex substrates. The properties shown in vitro for the crude extract suggest that it could have a positive effect on the protein metabolism of ruminants under intensive management and that negative side effects can be overcome by an optimized dosage. If the chemical nature of the active substance and its mechanism of action can be clarified, it may provide an alternative to replace critical synthetic feed additives (such as antibiotics) for high yielding dairy cows.     

A dose-response experiment evaluating the effects of oligofructose and inulin on nutrient digestibility, stool quality, and fecal protein catabolites in healthy adult dogs

In this experiment, three concentrations (0.3, 0.6, and 0.9% of diet, as-fed basis) of two fructans, oligofructose (OF) and inulin, were tested against a 0% supplemental fructan control. Seven ileal-cannulated adult female dogs were fed a meat-based, kibbled diet and assigned to treatments in a 7 x 7 Latin square design. Dietary supplementation of fructans had no effect on nutrient intakes or ileal digestibilities. Total-tract digestibilities of DM, OM, and CP decreased (P < 0.05) as a result of dietary OF and inulin supplementation. Dogs fed the control diet had a DM total-tract digestibility of 83.0%. The percentages of fecal DM for dogs fed the control and 0.3, 0.6, and 0.9% OF were 36.6, 33.3, 32.8, and 31.7%, respectively. When compared with the control, OF (P < 0.01) and inulin (P < 0.01) supplementation increased fecal ammonia concentrations. Higher fecal short-chain fatty acid (SCFA; P < 0.10) and isovalerate concentrations (P < 0.01) were noted for dogs fed both fructans. Total fecal SCFA for dogs fed the control diet and 0.3, 0.6, and 0.9% OF were 406.4, 529.9, 538.3, and 568.8 micromol/g of feces (DM basis), respectively. Dogs fed 0.3, 0.6, and 0.9% inulin had total fecal SCFA of 472.2, 468.8, and 471.5 micromol/g of feces (DM basis), respectively. Linear increases were observed in putrescine (P < 0.11), cadaverine (P < 0.07), spermidine (P < 0.12), and total amines (P < 0.05) in feces of dogs fed OF. Lower fecal phenol (P < 0.08) and total phenol (P < 0.04) concentrations occurred in dogs fed inulin, along with a linear decrease (P < 0.08) in total phenols with OF supplementation. Total fecal phenols for dogs fed the control, 0.3, 0.6, and 0.9% inulin were 3.03, 1.86, 1.97, and 2.23 micromol/g of feces (DM basis), respectively. Low-level dietary inclusion of inulin and OF positively affected indices known to be associated with gut health of the dog without seriously compromising nutrient digestibility or stool quality. Overall, the 0.9% OF treatment resulted in the best responses, including no adverse effect on nutrient intakes, ileal digestibilities, or stool quality, as well as increased fecal SCFA and decreased fecal phenols. The biological responses due to inulin were more variable.     

Effect of soybean husk supplementation on the fecal fermentation metabolites and microbiota of dogs

In vitro fermentation and in vivo feeding experiments were conducted to characterize the effects of soybean (Glycine max) husk on the fecal fermentation metabolites and microbiota of dogs. An in vitro fermentation study using feces from three Toy Poodle dogs (6.5 ± 3.5 months in age and 2.9 ± 0.4 kg in body weight) revealed that the fecal inoculum was able to ferment soybean husk (supplemented at 0.01 g/mL culture) and increased levels of short chain fatty acids (SCFA) and Bifidobacterium, irrespective of pre‐digestion of the husk by pepsin and pancreatin. In a feeding experiment, four Shiba dogs (7–48 months in age and 7.5 ± 1.7 kg in body weight) fed a commercial diet supplemented with 5.6% soybean husk showed an increase in SCFA, such as acetate and butyrate, and lactate, and a decrease in indole and skatole in the feces compared to those fed a 5.6% cellulose diet. Real‐time PCR assay showed that soybean husk supplementation stimulated the growth of lactobacilli, Clostridium cluster IV including Faecalibacterium prausnitzii, Clostridium cluster XIVa, Bacteroides‐Prevotella‐Porphyromonas group but inhibited the growth of Clostridium cluster XI. Both in vitro and in vivo experiments indicated that soybean husk supplementation improves gastrointestinal health through optimization of beneficial organic acid production and increase of beneficial bacteria. Therefore, soybean husk is suggested to be applicable as a functional fiber in the formulation of canine diets.     

Effects of supplemental fructooligosaccharides plus mannanoligosaccharides on immune function and ileal and fecal microbial populations in adult dogs.

The goal of this study was to examine whether supplemental fructooligosaccharides (FOS) plus mannanoligosaccharides (MOS) influenced immune function and ileal and fecal microbial populations of adult dogs. Eight adult dogs surgically fitted with ileal cannulas were used in a crossover design. Dogs were fed 200 g of a dry, extruded, kibble diet twice daily. At each feeding, dogs were dosed with either 1 g sucrose (placebo) or 2 g FOS plus 1 g MOS orally via gelatin capsule. Fecal, ileal, and blood samples were collected at the end of each 14-d period to measure microbial populations and immune characteristics. Treatment least squares means were compared using the GLM procedure of SAS. Supplementation of FOS plus MOS increased fecal bifidobacteria and fecal and ileal lactobacilli concentrations. Dogs fed FOS plus MOS also tended to have lower blood neutrophils and greater blood lymphocytes vs placebo. Serum, fecal, and ileal immunoglobulin concentrations were unchanged by treatment. Supplementation of FOS plus MOS beneficially altered indices of gut health by improving ileal and fecal microbial ecology. Supplementation of FOS plus MOS also altered immune function by causing a shift in blood immune cells.