Analyze Diet

Topic:Microbiome

The equine microbiome refers to the diverse community of microorganisms, including bacteria, fungi, and viruses, residing in various parts of a horse's body, such as the gastrointestinal tract, skin, and respiratory system. These microorganisms are involved in numerous physiological processes, including digestion, metabolism, and immune function. The composition and diversity of the microbiome can be influenced by factors such as diet, environment, and health status. Research in this area explores the characterization of the equine microbiome, its impact on health and disease, and potential applications in managing equine health. This page compiles peer-reviewed research studies and scholarly articles that investigate the composition, function, and implications of the microbiome in horses.
Comparison of the Fecal Microbiota of Horses with Intestinal Disease and Their Healthy Counterparts.
Veterinary sciences    June 17, 2021   Volume 8, Issue 6 doi: 10.3390/vetsci8060113
Park T, Cheong H, Yoon J, Kim A, Yun Y, Unno T.(1) Background: The intestinal microbiota plays an essential role in maintaining the host's health. Dysbiosis of the equine hindgut microbiota can alter the fermentation patterns and cause metabolic disorders. (2) Methods: This study compared the fecal microbiota composition of horses with intestinal disease and their healthy counterparts living in Korea using 16S rRNA sequencing from fecal samples. A total of 52 fecal samples were collected and divided into three groups: horses with large intestinal disease (n = 20), horses with small intestinal disease (n = 8), and healthy horses (n = 24). (...
The Equine Faecal Microbiota of Healthy Horses and Ponies in The Netherlands: Impact of Host and Environmental Factors.
Animals : an open access journal from MDPI    June 12, 2021   Volume 11, Issue 6 1762 doi: 10.3390/ani11061762
Theelen MJP, Luiken REC, Wagenaar JA, Sloet van Oldruitenborgh-Oosterbaan MM, Rossen JWA, Zomer AL.Several studies have described the faecal microbiota of horses and the factors that influence its composition, but the variation in results is substantial. This study aimed to investigate the microbiota composition in healthy equids in The Netherlands under standard housing and management conditions and to evaluate the effect of age, gender, horse type, diet, pasture access, the season of sampling and location on it. Spontaneously produced faecal samples were collected from the stall floor of 79 healthy horses and ponies at two farms. The validity of this sampling technique was evaluated in a ...
Establishment and assessment of an amplicon sequencing method targeting the 16S-ITS-23S rRNA operon for analysis of the equine gut microbiome.
Scientific reports    June 4, 2021   Volume 11, Issue 1 11884 doi: 10.1038/s41598-021-91425-7
Kinoshita Y, Niwa H, Uchida-Fujii E, Nukada T.Microbial communities are commonly studied by using amplicon sequencing of part of the 16S rRNA gene. Sequencing of the full-length 16S rRNA gene can provide higher taxonomic resolution and accuracy. To obtain even higher taxonomic resolution, with as few false-positives as possible, we assessed a method using long amplicon sequencing targeting the rRNA operon combined with a CCMetagen pipeline. Taxonomic assignment had > 90% accuracy at the species level in a mock sample and at the family level in equine fecal samples, generating similar taxonomic composition as shotgun sequencing. The...
Antimicrobial peptides in domestic animals and their applications in veterinary medicine.
Peptides    May 24, 2021   Volume 142 170576 doi: 10.1016/j.peptides.2021.170576
Valdez-Miramontes CE, De Haro-Acosta J, Aréchiga-Flores CF, Verdiguel-Fernández L, Rivas-Santiago B.Antimicrobial peptides (AMPs) are molecules with a broad-spectrum activity against bacteria, fungi, protozoa, and viruses. These peptides are widely distributed in insects, amphibians and mammals. Indeed, they are key molecules of the innate immune system with remarkable antimicrobial and immunomodulatory activity. Besides, these peptides have also shown regulatory activity for gut microbiota and have been considered inductors of growth performance. The current review describes the updated findings of antimicrobial peptides in domestic animals, such as bovines, goats, sheep, pigs, horses, cani...
An Oligosaccharide Rich Diet Increases Akkermansia spp. Bacteria in the Equine Microbiota.
Frontiers in microbiology    May 21, 2021   Volume 12 666039 doi: 10.3389/fmicb.2021.666039
Lindenberg FC, Lützhøft DO, Krych L, Fielden J, Kot W, Frøkiær H, van Galen G, Nielsen DS, Hansen AK.Some oligosaccharides induce growth of anti-inflammatory bacterial species and induce regulatory immunity in humans as well as animals. We have shown that the equine gut microbiota and the immune-microbial homeostasis largely stabilize within the first 50 days of life. Furthermore, we have previously established that certain bacterial species in the equine gut correlated with regulatory immunity. Accordingly, we hypothesized that an oligosaccharide rich diet fed to foals during the first 50 days would increase the abundance of bacterial species associated with regulatory immunity, and that thi...
Comparison of the Gut Microbiota of Jeju and Thoroughbred Horses in Korea.
Veterinary sciences    May 11, 2021   Volume 8, Issue 5 81 doi: 10.3390/vetsci8050081
Park T, Yoon J, Kim A, Unno T, Yun Y.(1) Background: The large intestine of horses is an anaerobic fermentative chamber filled with fibrolytic bacteria that play essential roles in digesting and absorbing nutrients for energy production. Although Jeju horses are a prominent local breed in Korea, few studies have investigated the gut microbiota of Jeju horses; (2) Methods: This study performed sequencing of V3 and V4 hypervariable regions of the partial 16S rRNA genes obtained from horse fecal samples and compared the gut microbiota between Jeju and Thoroughbred horses. Thirty and 24 fecal samples were obtained from Jeju and Thoro...
Effects of Gasterophilus pecorum infestation on the intestinal microbiota of the rewilded Przewalski’s horses in China.
PloS one    May 11, 2021   Volume 16, Issue 5 e0251512 doi: 10.1371/journal.pone.0251512
Hu D, Chao Y, Zhang B, Wang C, Qi Y, Ente M, Zhang D, Li K, Mok KM.Horse botflies have been a threat to the Przewalski's horses in the Kalamaili Nature Reserve in Xinjiang of China since their reintroduction to the original range. As larvae of these parasites could infest the intestine of a horse for months, they could interact with and alter the structure and composition of its intestinal microbiota, affecting adversely its health. Nonetheless, there are no such studies on the rewilded Przewalski's horses yet. For the first time, this study characterizes the composition of the intestinal microbiota of 7 rewilded Przewalski's horses infected severely by Gaste...
Effects of Pasture Grass, Silage, and Hay Diet on Equine Fecal Microbiota.
Animals : an open access journal from MDPI    May 7, 2021   Volume 11, Issue 5 doi: 10.3390/ani11051330
Zhu Y, Wang X, Deng L, Chen S, Zhu C, Li J.Diet is an important factor affecting intestinal microbiota in horses. Fecal microbiota is commonly used as a substitute for studying hindgut microbiota when investigating the relationship between intestinal microbial changes and host health. So far, no study has compared the difference between the fecal microbiota found in horses that are fed pasture grass, silage, and hay. The present study aims to characterize the fecal microbiota in horses that were exclusively on one of the three forage diets, and to analyze the potential impact of these forages, especially silage, on horse intestinal hea...
Sequential Modulation of the Equine Fecal Microbiota and Fibrolytic Capacity Following Two Consecutive Abrupt Dietary Changes and Bacterial Supplementation.
Animals : an open access journal from MDPI    April 29, 2021   Volume 11, Issue 5 1278 doi: 10.3390/ani11051278
Collinet A, Grimm P, Julliand S, Julliand V.In horses, abrupt changes from high-fiber (HF) to high-starch (HS) diets can affect the cecal and colonic microbiota. This study investigated modifications and recovery of fecal microbiota after two consecutive abrupt dietary changes. Twelve horses fed HF for 2 weeks were changed to HS for 5 days then returned to HF for 7 weeks. Six received lactic acid bacteria supplementation. Bacterial population diversity, structure, and activity, especially fibrolysis, were assessed to obtain an overview of alteration in hindgut microbiota. Two days after the abrupt change from HF to HS, the findings in f...
The equine hindgut as a reservoir of mobile genetic elements and antimicrobial resistance genes.
Critical reviews in microbiology    April 24, 2021   Volume 47, Issue 5 543-561 doi: 10.1080/1040841X.2021.1907301
Mitchell S, Bull M, Muscatello G, Chapman B, Coleman NV.Antibiotic resistance in bacterial pathogens is a growing problem for both human and veterinary medicine. Mobile genetic elements (MGEs) such as plasmids, transposons, and integrons enable the spread of antibiotic resistance genes (ARGs) among bacteria, and the overuse of antibiotics drives this process by providing the selection pressure for resistance genes to establish and persist in bacterial populations. Because bacteria, MGEs, and resistance genes can readily spread between different ecological compartments (e.g. soil, plants, animals, humans, wastewater), a "One Health" approach is need...
Nasopharyngeal Microbiomes in Donkeys Shedding Streptococcus equi Subspecies equi in Comparison to Healthy Donkeys.
Frontiers in veterinary science    April 22, 2021   Volume 8 645627 doi: 10.3389/fvets.2021.645627
Zhu Y, Chen S, Yi Z, Holyoak R, Wang T, Ding Z, Li J. subsp. equi () is the pathogen causing strangles, a highly infectious disease that can affect equids including donkeys of all ages. It can persistently colonize the upper respiratory tract of animals asymptomatically for years, which serves as a source of infection. Several strangles outbreaks have been reported in the donkey industry in China in the last few years and pose a great threat to health, production, and the welfare of donkeys. Nasopharyngeal swab samples for culture and PCR are used widely in strangles diagnosis. Additionally, microbiomes within and on the body are essential to ho...
Dynamic Changes in Microbiome Composition Following Mare’s Milk Intake for Prevention of Collateral Antibiotic Effect.
Frontiers in cellular and infection microbiology    April 21, 2021   Volume 11 622735 doi: 10.3389/fcimb.2021.622735
Kushugulova A, Löber U, Akpanova S, Rysbekov K, Kozhakhmetov S, Khassenbekova Z, Essex M, Nurgozhina A, Nurgaziyev M, Babenko D, Markó L....Probiotics and prebiotics are widely used for recovery of the human gut microbiome after antibiotic treatment. High antibiotic usage is especially common in children with developing microbiome. We hypothesized that dry Mare's milk, which is rich in biologically active substances without containing live bacteria, could be used as a prebiotic in promoting microbial diversity following antibiotic treatment in children. The present pilot study aims to determine the impacts of dry Mare's milk on the diversity of gut bacterial communities when administered during antibiotic treatment and throughout ...
Effects of Aleurone Supplementation on Glucose-Insulin Metabolism and Gut Microbiome in Untrained Healthy Horses.
Frontiers in veterinary science    April 12, 2021   Volume 8 642809 doi: 10.3389/fvets.2021.642809
Boshuizen B, Moreno de Vega CV, De Maré L, de Meeûs C, de Oliveira JE, Hosotani G, Gansemans Y, Deforce D, Van Nieuwerburgh F, Delesalle C.Aleurone, a layer of the bran fraction, is deemed to be responsible for the positive health effects associated with the consumption of whole-grain products. Studies on rodents, pigs, and humans report beneficial effects of aleurone in five main areas: the reduction of oxidative stress, immunomodulatory effects, modulation of energy management, digestive health, and the storage of vitamins and minerals. Our study is the first aleurone supplementation study performed in horses. The aim of this study was to investigate the effect of an increase in the dose levels of aleurone on the postprandial g...
The skin microbiota in equine pastern dermatitis: a case-control study of horses in Switzerland.
Veterinary dermatology    April 8, 2021   Volume 32, Issue 6 646-e172 doi: 10.1111/vde.12955
Kaiser-Thom S, Hilty M, Axiak S, Gerber V.Equine pastern dermatitis (EPD), a multifactorial syndrome, manifests as skin lesions of variable severity in the pastern area. Despite the widespread use of antibacterial therapy for treating this condition, little is known about the contributing bacteria. Objective: To investigate the bacterial skin microbiota in EPD-affected and unaffected (control) pasterns. Methods: Case-control study with 80 client-owned horses; each with at least one EPD-affected and one control pastern. Methods: Horses were grouped by the form of EPD (mild, exudative or proliferative), the assigned severity grade and t...
Understanding the Holobiont: Crosstalk Between Gut Microbiota and Mitochondria During Long Exercise in Horse.
Frontiers in molecular biosciences    April 8, 2021   Volume 8 656204 doi: 10.3389/fmolb.2021.656204
Mach N, Moroldo M, Rau A, Lecardonnel J, Le Moyec L, Robert C, Barrey E.Endurance exercise has a dramatic impact on the functionality of mitochondria and on the composition of the intestinal microbiome, but the mechanisms regulating the crosstalk between these two components are still largely unknown. Here, we sampled 20 elite horses before and after an endurance race and used blood transcriptome, blood metabolome and fecal microbiome to describe the gut-mitochondria crosstalk. A subset of mitochondria-related differentially expressed genes involved in pathways such as energy metabolism, oxidative stress and inflammation was discovered and then shown to be associa...
How Can Nutrition Help with Gastrointestinal Tract-Based Issues?
The Veterinary clinics of North America. Equine practice    April 7, 2021   Volume 37, Issue 1 63-87 doi: 10.1016/j.cveq.2020.12.007
Hesta M, Costa M.Many horses are fed differently than their wild ancestors. They often have limited access to pasture and are fed conserved forage and concentrates rich in starch and sugars, in only 2 meals per day. Feeding practices in contrast to natural feeding behavior can lead to gastrointestinal issues. Standard nutritional evaluation is warranted because of its important role in prevention and in treatment and management of diseases. When medical and nutritional treatments are combined, success rates are higher. New techniques to characterize equine microbiota have been used, allowing for microbiota man...
The Influence of Diet Change and Oral Metformin on Blood Glucose Regulation and the Fecal Microbiota of Healthy Horses.
Animals : an open access journal from MDPI    April 1, 2021   Volume 11, Issue 4 976 doi: 10.3390/ani11040976
Ericsson AC, Johnson PJ, Gieche LM, Zobrist C, Bucy K, Townsend KS, Martin LM, LaCarrubba AM.Common treatments for Equine Metabolic Syndrome (EMS) and associated conditions include removal from pasture and adoption of an all-hay diet. Pharmacological treatments for EMS include metformin, a biguanide antihyperglycemic agent also administered to people to help improve glucose tolerance and insulin sensitivity. Both treatments may work, at least partially, through the gut microbiota, yet little is known regarding these effects in the equine host. To determine the influence on the fecal microbiota of this diet change and administration of metformin, six healthy horses were removed from pa...
Multidimensional Approach for Investigating the Effects of an Antibiotic-Probiotic Combination on the Equine Hindgut Ecosystem and Microbial Fibrolysis.
Frontiers in microbiology    March 25, 2021   Volume 12 646294 doi: 10.3389/fmicb.2021.646294
Collinet A, Grimm P, Julliand S, Julliand V.The equine hindgut ecosystem is specialized in dietary fibers' fermentation to provide horses' energy and contribute to its health. Nevertheless, antibiotics are known to disrupt the hindgut microbiota, affecting the fibrolytic activity of bacteria and the intestinal immune balance, leading to diseases. This study used a general and comprehensive approach for characterizing the hindgut ecosystem of 9 healthy horses over 28 days in response to a 5-day challenge with oral trimethoprim-sulfadiazine (TMS), with a special emphasis on microbial fibrolytic activity and the host immune response. Hors...
Exodontia associated bacteremia in horses characterized by next generation sequencing.
Scientific reports    March 18, 2021   Volume 11, Issue 1 6314 doi: 10.1038/s41598-021-85484-z
Townsend KS, Johnson PJ, LaCarrubba AM, Martin LM, Ericsson AC.Bacteremia resulting from dental surgery is increasingly recognized as a health risk, especially in older and immunocompromised patients. Dentistry-associated bacteremia can lead to remote infections, as exemplified by valvular endocarditis. Emerging evidence points to a novel role played by oral cavity commensals in the pathogenesis of diabetes, respiratory disease, cardiovascular disease, and adverse pregnancy outcomes. Whether dental extraction, a commonly undertaken procedure in old horses, causes bacteremia has not been reported extensively. In a prospective clinical study using next gene...
Impact of Ambient Temperature Sample Storage on the Equine Fecal Microbiota.
Animals : an open access journal from MDPI    March 15, 2021   Volume 11, Issue 3 819 doi: 10.3390/ani11030819
Martin de Bustamante M, Plummer C, MacNicol J, Gomez D.Sample storage conditions are an important factor in fecal microbiota analyses in general. The objective of this study was to investigate the effect of sample storage at room temperature on the equine fecal microbiota composition. Fecal samples were collected from 11 healthy horses. Each sample was divided into 7 sealed aliquots. One aliquot was immediately frozen at -80 °C; the remaining aliquots were stored at room temperature (21 to 22 °C) with one transferred to the freezer at each of the following time points: 6, 12, 24, 48, 72 and 96 h. The Illumina MiSeq sequencer was used for high-th...
A Comparison of the Colonic Microbiome and Volatile Organic Compound Metabolome of Anoplocephala perfoliata Infected and Non-Infected Horses: A Pilot Study.
Animals : an open access journal from MDPI    March 9, 2021   Volume 11, Issue 3 doi: 10.3390/ani11030755
Slater R, Frau A, Hodgkinson J, Archer D, Probert C.Anoplocephala perfoliata is a common equine tapeworm associated with an increased risk of colic (abdominal pain) in horses. Identification of parasite and intestinal microbiota interactions have consequences for understanding the mechanisms behind parasite-associated colic and potential new methods for parasite control. A. perfoliata was diagnosed by counting of worms in the caecum post-mortem. Bacterial DNA was extracted from colonic contents and sequenced targeting of the 16S rRNA gene (V4 region). The volatile organic compound (VOC) metabolome of colonic contents was characterised using gas...
Long-term storage of feces at -80 °C versus -20 °C is negligible for 16S rRNA amplicon profiling of the equine bacterial microbiome.
PeerJ    March 9, 2021   Volume 9 e10837 doi: 10.7717/peerj.10837
Gavriliuc S, Stothart MR, Henry A, Poissant J.The development of next-generation sequencing technologies has spurred a surge of research on bacterial microbiome diversity and function. But despite the rapid growth of the field, many uncertainties remain regarding the impact of differing methodologies on downstream results. Sample storage temperature is conventionally thought to be among the most important factors for ensuring reproducibility across marker gene studies, but to date much of the research on this topic has focused on short-term storage in the context of clinical applications. Consequently, it has remained unclear if storage a...
The Fecal Bacterial Microbiota in Horses with Equine Recurrent Uveitis.
Animals : an open access journal from MDPI    March 9, 2021   Volume 11, Issue 3 doi: 10.3390/ani11030745
Martin de Bustamante M, Gomez D, MacNicol J, Hamor R, Plummer C.The objective of this study was to describe and compare the fecal bacterial microbiota of horses with equine recurrent uveitis (ERU) and healthy horses using next-generation sequencing techniques. Fecal samples were collected from 15 client-owned horses previously diagnosed with ERU on complete ophthalmic examination. For each fecal sample obtained from a horse with ERU, a sample was collected from an environmentally matched healthy control with no evidence of ocular disease. The Illumina MiSeq sequencer was used for high-throughput sequencing of the V4 region of the 16S rRNA gene. The relativ...
Effects of phenylbutazone alone or in combination with a nutritional therapeutic on gastric ulcers, intestinal permeability, and fecal microbiota in horses.
Journal of veterinary internal medicine    March 3, 2021   Volume 35, Issue 2 1121-1130 doi: 10.1111/jvim.16093
Whitfield-Cargile CM, Coleman MC, Cohen ND, Chamoun-Emanuelli AM, DeSolis CN, Tetrault T, Sowinski R, Bradbery A, Much M.Gastrointestinal (GI) injury and dysbiosis are adverse events associated with nonsteroidal anti-inflammatory drug (NSAID) use in horses. Phenylbutazone has been shown to alter GI barrier function both in vitro and ex vivo, but its effects on barrier function have not been assessed in vivo. In addition, the ability of nutritional therapeutics to prevent these changes is not known. Objective: Our objectives were to determine whether (a) phenylbutazone affected barrier function in vivo and (b) if phenylbutazone-induced GI injury could be ameliorated by the use of a nutritional therapeutic. Method...
Gut microbiota resilience in horse athletes following holidays out to pasture.
Scientific reports    March 3, 2021   Volume 11, Issue 1 5007 doi: 10.1038/s41598-021-84497-y
Mach N, Lansade L, Bars-Cortina D, Dhorne-Pollet S, Foury A, Moisan MP, Ruet A.Elite horse athletes that live in individual boxes and train and compete for hours experience long-term physical and mental stress that compromises animal welfare and alters the gut microbiota. We therefore assessed if a temporary period out to pasture with conspecifics could improve animal welfare and in turn, favorably affect intestinal microbiota composition. A total of 27 athletes were monitored before and after a period of 1.5 months out to pasture, and their fecal microbiota and behavior profiles were compared to those of 18 horses kept in individual boxes. The overall diversity and mic...
Faecal bacterial composition in horses with and without free faecal liquid: a case control study.
Scientific reports    February 26, 2021   Volume 11, Issue 1 4745 doi: 10.1038/s41598-021-83897-4
Lindroth KM, Dicksved J, Pelve E, Båverud V, Müller CE.Free faecal liquid (FFL) is a condition in horses which manifests as differential defecation of solid and liquid phases of faeces. The etiology of FFL is currently unknown, but deviances in the hindgut microbiota has been suggested to be of importance. The present study aimed to compare the faecal bacterial composition of farm-matched horses with (case, n = 50) and without (control, n = 50) FFL. Samples were collected at three different occasions. The V3 and V4 regions of the 16S rRNA gene were amplified and sequenced using Illumina sequencing. Also, samples were cultivated for detecti...
The pelvic flexure separates distinct microbial communities in the equine hindgut.
Scientific reports    February 22, 2021   Volume 11, Issue 1 4332 doi: 10.1038/s41598-021-83783-z
Reed KJ, Kunz IGZ, Scare JA, Nielsen MK, Turk PJ, Coleman RJ, Coleman SJ.As hindgut fermenters, horses are especially dependent on the microbiota residing in their cecum and large intestines. Interactions between these microbial populations and the horse are critical for maintaining gut homeostasis, which supports proper digestion. The current project was motivated to determine if any features of the fecal microbiota are informative of the microbial communities from the cecum, ventral colon, or dorsal colon. Digesta from the cecum, ventral colon, dorsal colon and feces were collected from 6 yearling miniature horses. Microbial DNA was isolated and the microbiota fr...
Comparison of Fecal Microbiota of Horses Suffering from Atypical Myopathy and Healthy Co-Grazers.
Animals : an open access journal from MDPI    February 15, 2021   Volume 11, Issue 2 doi: 10.3390/ani11020506
Wimmer-Scherr C, Taminiau B, Renaud B, van Loon G, Palmers K, Votion D, Amory H, Daube G, Cesarini C.Equine atypical myopathy (AM) is caused by hypoglycin A (HGA) and methylenecyclopropylglycine (MCPG) intoxication resulting from the ingestion of seeds or seedlings of some Acer tree species. Interestingly, not all horses pasturing in the same toxic environment develop signs of the disease. In other species, it has been shown that the intestinal microbiota has an impact on digestion, metabolism, immune stimulation and protection from disease. The objective of this study was to characterize and compare fecal microbiota of horses suffering from AM and healthy co-grazers. Furthermore, potential d...
Evaluation of the ocular surface mycobiota in clinically normal horses.
PloS one    February 4, 2021   Volume 16, Issue 2 e0246537 doi: 10.1371/journal.pone.0246537
Walsh ML, Meason-Smith C, Arnold C, Suchodolski JS, Scott EM.The eye is host to myriad bacterial, fungal, and viral organisms that likely influence ocular surface physiology in normal and diseased states. The ocular surface mycobiota of horses has not yet been described using NGS techniques. This study aimed to characterize the ocular surface fungal microbiota (mycobiota) in healthy horses in 2 environmental conditions (stalled versus pasture). Conjunctival swabs of both eyes were obtained from 7 adult stallions stabled in an open-air pavilion and 5 adult mares living on pasture. Genomic DNA was extracted from ocular surface swabs and sequenced using pr...
Effect of hay type on cecal and fecal microbiome and fermentation parameters in horses.
Journal of animal science    January 31, 2021   Volume 99, Issue 1 skaa407 doi: 10.1093/jas/skaa407
Sorensen RJ, Drouillard JS, Douthit TL, Ran Q, Marthaler DG, Kang Q, Vahl CI, Lattimer JM.The effect of hay type on the microbiome of the equine gastrointestinal tract is relatively unexplored. Our objective was to characterize the cecal and fecal microbiome of mature horses consuming alfalfa or Smooth Bromegrass (brome) hay. Six cecally cannulated horses were used in a split-plot design run as a crossover in two periods. The whole plot treatment was ad libitum access to brome or alfalfa hay fed over two 21-d acclimation periods with subplots of sampling location (cecum and rectum) and sampling hour. Each acclimation period was followed by a 24-h collection period where cecal and f...
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