The art of establishing mineral tolerances of dogs and cats.

For over six decades, nutritional science has provided well-developed, peer-reviewed nutrient recommendations to support the health of dogs and cats. These guidelines are updated based on new scientifically valid research and appropriate peer-review. Recent regulatory and scientific positions around health issues have resulted in strong opinions and desires for rapid regulatory action surrounding mineral nutrition, but with limited and conflicting scientific evidence. Pet Food Institute nutrition experts have come together to jointly author an article on the complexities of establishing mineral tolerances of dogs and cats to illustrate the limitations in defining mineral tolerances. This discussion covers how mineral requirements were determined, including the opportunities and pitfalls encountered. Scientific councils must review and clarify any proposed changes in conducting mineral nutrition research that might impact complete and balanced foods and surrounding regulations. It is important to clarify the multiple issues in mineral nutrition research and the necessity for thorough evaluation of data while avoiding arbitrary and potentially harmful guidelines.

Dogs and cats are living longer and healthier lives due, in part, to the scientific development of nutritional information. This information has allowed the building of many new types of foods, treats, and supplements that promote life, health, and enjoyment by the pet. There are several organizations that have provided helpful reviews of nutritional data through scientific councils that help identify safe and healthy criteria for all pet food products. These are readily available for those who want to learn more about pet nutrition. For many nutrients, there is a large database of information to help build products. Nutrients that are called macro- or micro-/trace minerals (e.g., sodium, potassium, zinc, copper, etc.) often have more limited information. Recently, strong opinions about pet health as related to mineral nutrition have been shared and robustly communicated without adequate scientific research to support the hypotheses. This has led to misinformation, concerns, and fear. To safeguard the health of companion animals and provide assistance to regulatory bodies regarding the nutritional welfare of dogs and cats, scientific panels have come together from industry, government, and academia to review, approve, and challenge nutritional guidelines. This overview provides the reader context into the rigor needed to establish safe mineral tolerances for dogs and cats.

Copper metabolism and its implications for canine nutrition.

Canine copper nutrition has received increased attention due to recent reports of apparent copper-associated hepatitis in the USA and European Union. In order to properly address the need to modify the U.S. National Research Council and Association of American Feed Control Officials canine copper recommendations that will have implications for all dogs, it is important to understand the complexities of copper metabolism, confounding variables affecting copper status, and the available research on this topic in dogs. Recent trends in consumer preference for dog diets, supplements, and functional treats introduce another layer of complexity, as most ingredients used in these formulations provide vastly different proportions of essential nutrients, thus resulting in great variation in nutrient profiles available to the animal. Although controlled research addressing copper metabolism in dogs is limited, there are lessons to be learned from other monogastric species as well as canine case studies that can provide a base for increasing knowledge to address this issue. Copper metabolism and status in animals is affected by a multitude of factors including absorption, storage, excretion, and nutrient interactions. Given its vital role in many physiological processes, it is important that both nutritional deficiencies and toxicities be avoided. Additionally, another challenge for proper copper nutrition in dogs is the known genetic predispositions of some breeds for copper storage and excretion abnormalities. Therefore, it is imperative that veterinarians, nutritionists, and pet food manufacturers collaborate with the shared goal of providing dog food options that supply the essential nutrients at adequate concentrations to support an active and healthy life. Many questions remain regarding copper metabolism and proper diet formulation for dogs. Future research efforts should focus on discovering reliable, non-invasive methods for evaluating canine copper status, a deeper understanding of genetic predispositions of certain breeds, increased knowledge of copper contributions from various ingredients, and the role of unpredictable physiological stressors on copper metabolism.

Perspective: Assessing Tolerance to Nondigestible Carbohydrate Consumption.

Human intestinal enzymes do not hydrolyze nondigestible carbohydrates (NDCs), and thus, they are not digested and absorbed in the small intestine. Instead, NDCs are partially to completely fermented by the intestinal microbiota. Select NDCs are associated with health benefits such as laxation and lowering of blood cholesterol and glucose. NDCs provide functional attributes to processed foods, including sugar or fat replacers, thickening agents, and bulking agents. Additionally, NDCs are incorporated into processed foods to increase their fiber content. Although consumption of NDCs can benefit health and contribute functional characteristics to foods, they can cause gastrointestinal symptoms, such as flatulence and bloating. As gastrointestinal symptoms negatively affect consumer well-being and their acceptance of foods containing NDC ingredients, it is crucial to consider tolerance when designing food products and testing their physiological health benefits in clinical trials. This perspective provides recommendations for the approach to assess gastrointestinal tolerance to NDCs, with a focus on study design, population criteria, intervention, comparator, and outcome. Special issues related to studies in children and implications for stakeholders are also discussed. It is recommended that the evaluation of gastrointestinal tolerance to NDCs be conducted in randomized, blinded, controlled crossover studies using standard gastrointestinal questionnaires, with attention to study participant background diets, health status, lifestyle, and medications.

Effect of Dietary Inulin Supplementation on the Gut Microbiota Composition and Derived Metabolites of Individuals Undergoing Hemodialysis: A Pilot Study.

OBJECTIVE: The prebiotic fiber inulin has been studied in individuals undergoing hemodialysis (HD) due to its ability to reduce gut microbiota-derived uremic toxins. However, studies examining the effects of inulin on the gut microbiota and derived metabolites are limited in these patients. We aimed to assess the impact of a 4-week supplementation of inulin on the gut microbiota composition and microbial metabolites of patients on HD. DESIGN AND METHODS: In a randomized, double-blind, placebo-controlled, crossover study, twelve HD patients (55 ± 10 y, 50% male, 58% Black American, BMI 31.6 ± 8.9 kg/m2, 33% diabetes mellitus) were randomized to consume inulin [10 g/d for females; 15 g/d for males] or maltodextrin [6 g/d for females; 9 g/d for males] for 4 weeks, with a 4-week washout period. We assessed the fecal microbiota composition, fecal metabolites (short-chain fatty acids (SCFA), phenols, and indoles), and plasma indoxyl sulfate and p-cresyl sulfate. RESULTS: At baseline, factors that explained the gut microbiota variability included BMI category and type of phosphate binder prescribed. Inulin increased the relative abundance of the phylum Verrucomicrobia and its genus Akkermansia (P interaction = 0.045). Inulin and maltodextrin resulted in an increased relative abundance of the phylum Bacteroidetes and its genus Bacteroides (P time = 0.04 and 0.03, respectively). Both treatments increased the fecal acetate and propionate (P time = 0.032 and 0.027, respectively), and there was a trend toward increased fecal butyrate (P time = 0.06). Inulin did not reduce fecal p-cresol or indoles, or plasma concentrations of p-cresyl sulfate or indoxyl sulfate. CONCLUSIONS: A 4-week supplementation of inulin did not lead to major shifts in the fecal microbiota and gut microbiota-derived metabolites. This may be due to high variability among participants and an unexpected increase in fecal excretion of SCFA with maltodextrin. Larger studies are needed to determine the effects of prebiotic fibers on the gut microbiota and clinical outcomes to justify their use in patients on HD.

ASAS-NANP SYMPOSIUM: RUMINANT/NONRUMINANT FEED COMPOSITION: Challenges and opportunities associated with creating large feed composition tables.

Traditional feed composition tables have been a useful tool in the field of animal nutrition throughout the last 70 yr. The objective of this paper is to discuss the challenges and opportunities associated with creating large feed ingredient composition tables. This manuscript will focus on three topics discussed during the National Animal Nutrition Program (NANP) Symposium in ruminant and nonruminant nutrition carried out at the American Society of Animal Science Annual Meeting in Austin, TX, on July 11, 2019, namely: 1) Using large datasets in feed composition tables and the importance of standard deviation in nutrient composition as well as different methods to obtain accurate standard deviation values, 2) Discussing the importance of fiber in animal nutrition and the evaluation of different methods to estimate fiber content of feeds, and 3) Description of novel feed sources, such as insects, algae, and single-cell protein, and challenges associated with the inclusion of such feeds in feed composition tables. Development of feed composition tables presents important challenges. For instance, large datasets provided by different sources tend to have errors and misclassifications. In addition, data are in different file formats, data structures, and feed classifications. Managing such large databases requires computers with high processing power and software that are also able to run automated procedures to consolidate files, to screen out outlying observations, and to detect misclassified records. Complex algorithms are necessary to identify misclassified samples and outliers aimed to obtain accurate nutrient composition values. Fiber is an important nutrient for both monogastrics and ruminants. Currently, there are several methods available to estimate the fiber content of feeds. However, many of them do not estimate fiber accurately. Total dietary fiber should be used as the standard method to estimate fiber concentrations in feeds. Finally, novel feed sources are a viable option to replace traditional feed sources from a nutritional perspective, but the large variation in nutrient composition among batches makes it difficult to provide reliable nutrient information to be tabulated. Further communication and cooperation among different stakeholders in the animal industry are required to produce reliable data on the nutrient composition to be published in feed composition tables.

Laxative effects of wheat bran and psyllium: Resolving enduring misconceptions about fiber in treatment guidelines for chronic idiopathic constipation.

BACKGROUND: Treatment guidelines for chronic idiopathic constipation (CIC) recommend an "increase in fiber intake" as a first-line therapy, but most epidemiologic studies fail to support an association between a high-fiber diet and a reduced risk of constipation. Furthermore, randomized controlled clinical studies show that most isolated fibers (e.g., supplements) are not different from placebo for a laxative effect, and several may be constipating. OBJECTIVES: The objective of this review was to compare the effects of two isolated fibers, coarse wheat bran and psyllium, on stool output and stool water content in patients with CIC. This review will also address misconceptions about fiber that are perpetuated by treatment guidelines. DATA SOURCES: A comprehensive literature review was conducted with the use of the Scopus, SciFinder, and PubMed scientific databases, limited to the previous 50 years (1968-2018; latest date included, December 31, 2018). CONCLUSIONS: In patients with CIC, nonfermented gel-forming psyllium was 3.4 times more effective than insoluble wheat bran for increasing stool output. Both psyllium and coarse wheat bran increased stool water content, a stool-softening effect, but finely ground wheat bran decreased stool water content, a stool-hardening effect. IMPLICATIONS FOR PRACTICE: It is a misconception that dietary fiber and all isolated fibers provide a laxative effect in patients with CIC. Our analysis suggests that treatment guidelines for CIC should make specific evidence-based recommendations as it pertains to fiber. To do otherwise takes the risk of perpetuating myth and misunderstanding and depriving patients of an effective therapy for CIC. A generic recommendation to "increase fiber intake" is akin to a recommendation to "increase pill intake" without regard to therapeutic or adverse effects.

Perspective: Physiologic Importance of Short-Chain Fatty Acids from Nondigestible Carbohydrate Fermentation.

In recent years, it has become increasingly obvious that dietary fiber or nondigestible carbohydrate (NDC) consumption is critical for maintaining optimal health and managing symptoms of metabolic disease. In accordance with this, the US FDA released its first official definition of dietary fiber in 2016 for regulation of Nutrition and Supplement Facts labels. Included in this definition is the requirement of an isolated or synthetic NDC to produce an accepted physiologic health benefit, such as improved laxation or reduced fasting cholesterol concentrations, upon consumption. Even though NDC fermentation and production of short-chain fatty acids elicit many physiologic effects, including serving as a source of energy for colonocytes, curbing glycemic response and satiety, promoting weight loss, enhancing mineral absorption, reducing systemic inflammation, and improving intestinal health, the process of fermentation is not considered a physiologic endpoint. Instead, expensive and laborious clinical trials must be conducted and an accepted physiologic benefit observed. In this review, we discuss the physiologic importance of NDC fermentation through extensive examination of clinical evidence and propose that the degree of fermentability of an NDC, rather than the endpoints of a clinical trial, may be appropriate for classifying it as a dietary fiber.

Behavioral response to fiber feedingis cohort-dependent and associated with gut microbiota composition in mice.

Recent data has supported a role for the gut microbiota in improving cognition and shaping behavior. Here, we assessed whether pectin, a soluble, fermentable fiber, could enhance learning and memory in mice. Two cohorts of young male C57Bl/6 J mice, C1 (n = 20) and C2 (n = 20), were obtained from Jackson Laboratory and randomized to semi-purified AIN-93 M diets containing 5% pectin (n = 10) or cellulose (n = 10). After 16 weeks, learning and memory was assessed by Morris Water Maze (MWM) and microbiota composition was analyzed by 16S rRNA sequencing. Despite identical treatment, we observed differences in learning and memory abilities between cohorts, along with distinct microbiotas. In C1, pectin-fed mice spent a higher percentage of time in the target quadrant at the 24-h probe trial of the MWM versus cellulose-fed mice; in C2, no effect of pectin was observed. In both cohorts, UniFrac distance revealed significant differences in gut microbial communities between cellulose-fed and pectin-fed mice. UniFrac analysis also revealed significantly different bacterial communities between cohorts. Further analysis demonstrated that the microbial genera Oscillospira, Bilophila, and Peptostreptococcoceae were more abundant in C1 versus C2, and positively associated with distance from the platform during the 24-h probe test. These data support previous findings that differences in the gut microbiota may play a role in host response to a dietary intervention and could partly explain irreproducibility in psychological and behavioral experiments. Further research is needed to determine if a causal relationship exists.

Critical Factors in Determining Fiber Content of Feeds and Foods and Their Ingredients.

Background: Because of its chemical complexity, the estimation of dietary fiber content of feed and food materials is a difficult analytical challenge. Three major fiber analyses are conducted routinely in the United States including crude fiber (CF), detergent fiber, and total dietary fiber (TDF). Objective: Factors crucial to the successful measurement of dietary fibers are described and suggestions provided as to how to overcome potential analytical problems within assays. Methods: An accounting of methodological details that result in variation in fiber concentration values is presented along with suggestions as to how to decrease the variation. Results: CF analysis remains in use in the livestock feed and pet food industries for nutrition labeling purposes in spite of the fact that the analysis does not separate mammalian enzyme-digestible from indigestible carbohydrate components, and values obtained are usually 30-50% of the actual dietary fiber concentration. Detergent fiber methods quantify the insoluble dietary fibers (IDF) accurately, but not the soluble dietary fiber (SDF) components. TDF methods account for intrinsic and intact fibers, isolated and extracted fibers, and synthetic fibers found in feed and food ingredients and complete diet matrixes. Conclusions: The CF procedure should be abandoned as it fails to quantify fiber properly. Detergent analyses quantify IDF. TDF methods quantify both IDF and SDF. Highlights: Accurate dietary fiber quantification is essential given the role of fiber in health and well-being of animals and humans.

Considerations for best practices in studies of fiber or other dietary components and the intestinal microbiome.

A 2-day workshop organized by the National Institutes of Health and U.S. Department of Agriculture included 16 presentations focused on the role of diet in alterations of the gastrointestinal microbiome, primarily that of the colon. Although thousands of research projects have been funded by U.S. federal agencies to study the intestinal microbiome of humans and a variety of animal models, only a minority addresses dietary effects, and a small subset is described in sufficient detail to allow reproduction of a study. Whereas there are standards being developed for many aspects of microbiome studies, such as sample collection, nucleic acid extraction, data handling, etc., none has been proposed for the dietary component; thus this workshop focused on the latter specific point. It is important to foster rigor in design and reproducibility of published studies to maintain high quality and enable designs that can be compared in systematic reviews. Speakers addressed the influence of the structure of the fermentable carbohydrate on the microbiota and the variables to consider in design of studies using animals, in vitro models, and human subjects. For all types of studies, strengths and weaknesses of various designs were highlighted, and for human studies, comparisons between controlled feeding and observational designs were discussed. Because of the lack of published, best-diet formulations for specific research questions, the main recommendation is to describe dietary ingredients and treatments in as much detail as possible to allow reproduction by other scientists.

Phenological changes in bamboo carbohydrates explain the preference for culm over leaves by giant pandas (Ailuropoda melanoleuca) during spring.

Seasonal changes in the foodscape force herbivores to select different plant species or plant parts to meet nutritional requirements. We examined whether the search for calorie-rich carbohydrates explained giant panda's selection for bamboo culm over leaves during spring. Leaves and culms were collected from four Phyllostachys bamboos (P. aurea, P. aureosulcata, P. glauca, and P. nuda) once per month over 18-27 months. Monthly changes in annual plant part nutrients were examined, and compared to seasonal foraging behaviors of captive giant pandas. Although total fiber was greater (p<0.0001) in culm (85.6 ± 0.5%) than leaves (55.3 ± 0.4%) throughout the year, culm fiber was at its lowest in spring (79-85%) when culm selection by giant pandas exceeded 70% of their overall diet. Culm starch also was greatest (p = 0.044) during spring (5.5 ± 1.1%) and 2.5-fold the percentage of starch in leaves (2.2 ± 0.6%). The free sugars in spring culm consisted of a high proportion of glucose (35%) and fructose (47%), whereas sucrose made up 42% of the total free sugar content of spring leaves. Bound sugars in culm consisted of 60% glucose and 38% xylose likely representative of hemicellulose. The concentrations of bound sugars (hemicelluloses) in spring culms (543.7 ± 13.0 mg/g) was greater (p<0.001) than in leaves (373.0 ± 14.8 mg/g). These data help explain a long-standing question in giant panda foraging ecology: why consume the plant part with the lowest protein and fat during the energetically intensive spring breeding season? Giant pandas likely prefer spring culm that contains abundant mono- and polysaccharides made more bioavailable as a result of reduced fiber content. These data suggest that phenological changes in bamboo plant part nutrition drive foraging decisions by giant pandas.

In Vitro Digestion and Fermentation of Microencapsulated Tributyrin for the Delivery of Butyrate.

Butyrate possesses negative sensory qualities and is most effectively utilized in the intestine to provide energy to the colonocyte for the maintenance of intestinal health. Butyrate has also shown promise in the treatment of intestinal disorders and diseases such as short bowel syndrome, inflammatory bowel disease, and colon cancer. To modify sensory properties, intestinal release, and butyrate production capabilities, tributyrin (TB) was microencapsulated in whey protein isolate (WPI)-based and gamma-cyclodextrin (GC)-based materials. Using an in vitro digestion and fermentation model, microcapsules containing TB were monitored for their release and production of butyrate in vitro. All samples containing TB showed limited butyrate release (<5%) during oral and gastric stages. In the small intestinal phase, all microcapsules containing TB released approximately 75% of their total butyrate with no significant differences (P > 0.05) across formulations. During the fermentation phase, GC-based microcapsules produced significantly more butyrate (P < 0.001) on a molar basis than all WPI-based microcapsules. Butyrate production increased significantly (P < 0.001) over each time interval with GC-based microcapsules having the highest during the 12 h of fermentation. The GC-based TB encapsulation systems were able to effectively deliver butyrate to the small intestine and generate butyrate in the large intestine. These microcapsules may, therefore, be beneficial for the maintenance of intestinal health and improvement of disease states across all areas of the gastrointestinal tract.

Innovations in Canine and Feline Nutrition: Technologies for Food and Nutrition Assessment.

Pet owners have increasing concerns about the nutrition of their pets, and they desire foods and treats that are safe, traceable, and of high nutritive value. To meet these high expectations, detailed chemical composition characterization of ingredients well beyond that provided by proximate analysis will be required, as will information about host physiology and metabolism. Use of faster and more precise analytical methodology and novel technologies that have the potential to improve pet food safety and quality will be implemented. In vitro and in vivo assays will continue to be used as screening tools to evaluate nutrient quality and adequacy in novel ingredients prior to their use in animal diets. The use of molecular and high-throughput technologies allows implementation of noninvasive studies in dogs and cats to investigate the impact of dietary interventions by using systems biology approaches. These approaches may further improve the health and longevity of pets.

In Vitro Fermentation of Xylooligosaccharides Produced from Miscanthus × giganteus by Human Fecal Microbiota.

Purified xylooligosaccharides from Miscanthus × giganteus (M×G XOS) were used in an in vitro fermentation experiment inoculated with human fecal microbiota. A commercial XOS product and pectin were used as controls. Decreases in pH by 2.3, 2.4, and 2.0 units and production of short-chain fatty acids (SCFA; acetic acid, 7764.2, 6664.1, and 6387.9 µmol/g; propionic acid, 1006.7, 1089.5, and 661.5 µmol/g; and butyric acid, 955.5, 1252.9, and 917.7 µmol/g) were observed in M×G XOS, commercial XOS, and pectin medium after 12 h of fermentation, respectively. Titers of Bifidobacterium spp., Lactobacillus spp., and Escherichia coli increased when fed all three substrates as monitored by qPCR. There was no significant trend for Clostridium perfringens. During fermentation, M×G XOS was statistically equivalent in performance to the commercial XOS sample as measured by culture acidification and growth of health-promoting bacteria and resulted in the highest SCFA production among the three substrates.

Compositional Analysis of Whole Grains, Processed Grains, Grain Co-Products, and Other Carbohydrate Sources with Applicability to Pet Animal Nutrition.

Our objective was to measure the proximate, starch, amino acid, and mineral compositions of grains, grain co-products, and other carbohydrate sources with potential use in pet foods. Thirty-two samples from barley (barley flake, cut barley, ground pearled barley, malted barley, whole pearled barley, pearled barley flakes, and steamed rolled barley); oats (groats, ground oatmeal, ground steamed groats, instant oats, oat bran, oat fiber, oat flour, quick oats, regular rolled oats, steamed rolled oat groats, and steel cut groats); rice (brown rice, polished rice, defatted rice bran, and rice flour); and miscellaneous carbohydrate sources (canary grass seed, hulled millet, whole millet, quinoa, organic spelt hull pellets, potato flake, sorghum, whole wheat, and whole yellow corn) were analyzed. Crude protein, amino acid, fat, dietary fiber, resistant starch, and mineral concentrations were highly variable among the respective fractions (i.e., barley flake vs. malted barley vs. steamed rolled barley) as well as among the various grains (i.e., barley flake vs. brown rice vs. canary grass seed). These ingredients not only provide a readily available energy source, but also a source of dietary fiber, resistant starch, essential amino acids, and macrominerals for pet diets.

Agave Inulin Supplementation Affects the Fecal Microbiota of Healthy Adults Participating in a Randomized, Double-Blind, Placebo-Controlled, Crossover Trial.

BACKGROUND: Prebiotics resist digestion, providing fermentable substrates for select gastrointestinal bacteria associated with health and well-being. Agave inulin differs from other inulin type fibers in chemical structure and botanical origin. Preclinical animal research suggests these differences affect bacterial utilization and physiologic outcomes. Thus, research is needed to determine whether these effects translate to healthy adults. OBJECTIVE: We evaluated agave inulin utilization by the gastrointestinal microbiota by measuring fecal fermentative end products and bacterial taxa. METHODS: A randomized, double-blind, placebo-controlled, 3-period, crossover trial was undertaken in healthy adults (n = 29). Participants consumed 0, 5.0, or 7.5 g agave inulin/d for 21 d with 7-d washouts between periods. Participants recorded daily dietary intake; fecal samples were collected during days 16-20 of each period and were subjected to fermentative end product analysis and 16S Illumina sequencing. RESULTS: Fecal Actinobacteria and Bifidobacterium were enriched (P < 0.001) 3- and 4-fold after 5.0 and 7.5 g agave inulin/d, respectively, compared with control. Desulfovibrio were depleted 40% with agave inulin compared with control. Agave inulin tended (P < 0.07) to reduce fecal 4-methyphenol and pH. Bivariate correlations revealed a positive association between intakes of agave inulin (g/kcal) and Bifidobacterium (r = 0.41, P < 0.001). Total dietary fiber intake (total fiber plus 0, 5.0, or 7.5 g agave inulin/d) per kilocalorie was positively associated with fecal butyrate (r = 0.30, P = 0.005), tended to be positively associated with Bifidobacterium (r = 0.19, P = 0.08), and was negatively correlated with Desulfovibrio abundance (r = -0.31, P = 0.004). CONCLUSIONS: Agave inulin supplementation shifted the gastrointestinal microbiota composition and activity in healthy adults. Further investigation is warranted to determine whether the observed changes translate into health benefits in human populations. This trial was registered at clinicaltrials.gov as NCT01925560.

Modulation of the faecal microbiome of healthy adult dogs by inclusion of potato fibre in the diet.

Inclusion of fermentable fibres in the diet can have an impact on the hindgut microbiome and provide numerous health benefits to the host. Potato fibre (PF), a co-product of potato starch isolation, has a favourable chemical composition of pectins, resistant and digestible starch, cellulose, and hemicelluloses. The objective of the present study was to evaluate the effect of increasing dietary PF concentrations on the faecal microbiome of healthy adult dogs. Fresh faecal samples were collected from ten female dogs with hound bloodlines (6·13 (SEM 0·17) years; 22·0 (SEM 2·1) kg) fed five test diets containing graded concentrations of PF (0, 1·5, 3, 4·5 or 6% as-fed; Roquette Frères) in a replicated 5 × 5 Latin square design. Extraction of DNA was followed by amplification of the V4-V6 variable region of the 16S rRNA gene using barcoded primers. Sequences were classified into taxonomic levels using Basic Local Alignment Search Tool (BLASTn) against a curated GreenGenes database. Inclusion of PF increased (P< 0·05) the faecal proportions of Firmicutes, while those of Fusobacteria decreased (P< 0·05). Similar shifts were observed at the genus level and were confirmed by quantitative PCR (qPCR) analysis. With increasing concentrations of PF, faecal proportions of Faecalibacterium increased (P< 0·05). Post hoc Pearson's correlation analysis showed positive (P< 0·05) correlations with Bifidobacterium spp. and butyrate production and Lactobacillus spp. concentrations. Overall, increases in the proportion of Faecalibacterium (not Lactobacillus/Bifidobacterium, as confirmed by qPCR analysis) and faecal SCFA concentrations with increasing dietary PF concentrations suggest that PF is a possible prebiotic fibre.

Fiber supplementation influences phylogenetic structure and functional capacity of the human intestinal microbiome: follow-up of a randomized controlled trial.

BACKGROUND: In our published randomized, double-blind, placebo-controlled, 3-period crossover trial, healthy adult men (n = 21) consumed bars containing no supplemental fiber (placebo; NFC), polydextrose (21 g/d), and soluble corn fiber (SCF; 21 g/d) for 21 d each. Fecal specimens were collected between days 16 and 21 for fermentative end-product analysis and 16S ribosomal RNA bacterial gene amplification for bacterial taxa identification. Fiber supplementation decreased fecal putrefaction compounds and shifted abundances of several bacterial taxa. OBJECTIVE: The objective was to perform whole-genome shotgun 454 pyrosequencing on the same fecal specimens collected in that clinical trial to obtain comprehensive fecal bacterial genome sequencing coverage and explore the full range of bacterial genetic information in the fecal microbiome, thereby using a systematic approach to study the impact of dietary fiber supplementation on fecal metabolites, bacterial taxa, and bacterial metagenomes. DESIGN: Fecal samples were subjected to whole-genome shotgun 454 pyrosequencing to identify both fecal bacterial populations present and their functional genetic capacity. RESULTS: Whole-genome shotgun sequencing results revealed that fiber consumption shifted the Bacteroidetes:Firmicutes ratio, increasing the relative abundance of Bacteroidetes 12 ± 2% and 13 ± 2% with polydextrose and SCF, respectively, compared with NFC. Bivariate correlations showed a positive correlation between the Bacteroidetes:Firmicutes ratio and total dietary fiber intake but not body mass index. Principal coordinates analysis of Bray-Curtis distances indicated that bacterial gene composition was more similar in participants consuming fibers (polydextrose and SCF combined) in comparison with NFC. Shifts in bacterial gene abundances after polydextrose and SCF supplementation included genes associated with carbohydrate, amino acid, and lipid metabolism, as well as metabolism of cofactors and vitamins. CONCLUSION: This study conveys novel information about the impact of dietary fiber supplementation on the phylogenetic structure and functional capacity of the fecal microbiome of healthy adults.

In vitro hydrolytic digestion, glycemic response in dogs, and true metabolizable energy content of soluble corn fibers.

The objective of this research was to measure in vitro hydrolytic digestion, glycemic and insulinemic responses in dogs, and true ME (TMEn) content of select soluble corn fibers (SCF) in roosters. The first generation (G1) SCF included hydrochloric acid-treated corn syrup (G1-CS-HCl), an SCF with an increased total dietary fiber (TDF) content (G1-SCF-HCl), an SCF that was spray-dried (G1-SCF-SD), and a hydrogenated SCF (G1-SCF-hydrog). The second generation (G2) SCF included those prepared using phosphoric acid catalyzation in both a liquid [G2-SCF-phos (Lq)] and powder [G2-SCF-phos (Pw)] form, and SCF that were prepared using hydrochloric acid catalyzation in both a liquid [G2-SCF-HCl (Lq)] and powder [G2-SCF-HCl (Pw)] form. Also, in the G2 set of samples were SCF prepared using the same method, but in 3 separate batches, all of which contained 70% TDF and 15% sugars. Two were in liquid form [G2-SCF-phos+HCl (Lq1)] and [G2-SCF-phos+HCl (Lq2)], and one in powder form ([G2-SCF-phos+HCl (Pw)]. A lower sugar form (80% TDF and 5% sugar) of SCF was also evaluated (G2-SCF-low sugar). Glucose was the major free sugar and bound monosaccharide in all SCF except for G1-SCF-hydrog that had greater concentrations of sorbitol. All SCF had intermediate to low amounts of monosaccharides released as a result of in vitro hydrolytic digestion, with glucose being the primary sugar component released. The G1-SCF were more digestible in vitro (approximately 50%) compared to G2-SCF (approximately 32%). All SCF had attenuated glycemic responses in adult dogs compared to a maltodextrin control (P < 0.05). The G2-SCF, on average, had lower glycemic responses and TMEn values in roosters than G1-SCF. All SCF had low free sugar concentrations with varying degrees of resistance to digestion, reduced caloric content, and attenuated glycemic and insulinemic responses in adult dogs. These ingredients are potential candidates for inclusion in reduced calorie and low glycemic canine diets.