Analyze Diet

Topic:Biochemistry

The study of biochemistry in horses encompasses the chemical processes and substances that occur within equine organisms. This field investigates the molecular interactions and pathways that are fundamental to horse physiology, including metabolism, enzyme activity, and genetic expression. Key areas of interest include the examination of metabolic disorders, nutrient absorption, and the biochemical basis of muscle function and energy production. Researchers utilize biochemical analysis to understand health and disease mechanisms in horses, contributing to the development of diagnostic tools and therapeutic strategies. This page gathers peer-reviewed studies and scholarly articles that explore various biochemical processes and their implications for equine health and performance.
The effect of long term astaxanthin supplementation on the antioxidant status of racing Arabian horses – preliminary study.
Scientific reports    November 14, 2024   Volume 14, Issue 1 27991 doi: 10.1038/s41598-024-77732-9
Giercuszkiewicz-Hecold B, Kulka M, Czopowicz M, Wilczak J, Szarska E, Strzelec K, Grzeczka A, Graczyk S, Hryniszyn A, Mularczyk M, Marycz K....Astaxanthin due to its strong antioxidant activity is believed to reduce oxidative stress and therefore is considered as feed additive in pathological conditions and also for the athletes. It is promoted by several equine web portals, however, data supporting that concept in horses is limited. Thus, the aim of this study was to evaluate the effect of astaxanthin supplementation on the parameters of oxidative status in 3 years old, racing Arabian horses during long term observation and the changes related to a single training session of high intensity. Six horses were supplemented with astaxant...
Fibrolytic efficiency of the large intestine microbiota may benefit running speed in French trotters: A pilot study.
Physiological reports    November 13, 2024   Volume 12, Issue 21 e70110 doi: 10.14814/phy2.70110
Vasseur M, Lepers R, Langevin N, Julliand S, Grimm P.This pilot study sought to explore the contribution of the large intestine microbiota to energy metabolism and exercise performance through its ability to degrade fibers into short-chain fatty acids (SCFAs). To investigate this, a correlational study was carried out on athlete horses under the same management conditions. Fecal microbiota diversity and composition, fibrolytic efficiency and SCFAs were analyzed. An incremental running test was carried out to estimate the maximal running speed (MRS) of the horses, and blood samples were taken to measure energy metabolism parameters. MRS was posit...
Preliminary Functional Analysis of the Gut Microbiome in Colic Horses.
Animals : an open access journal from MDPI    November 10, 2024   Volume 14, Issue 22 3222 doi: 10.3390/ani14223222
Thomson P, Garrido D, Santibáñez R, Lara F.The gut microbiome plays a critical role in maintaining horse health, influencing digestion, immunity, and overall well-being. However, in certain conditions like colic, there is evidence of significant alterations in the microbial community. To analyze the composition of the fecal microbiome and the enriched predicted metabolic functions of horses with colic compared to a control group, 14 horses with colic and 14 control horses were recruited. From a stool sample, DNA extraction was carried out for subsequent 16S rRNA metagenomic analysis. The composition of the microbiome was analyzed from ...
Peptidomic Analysis of Potential Bioactive Peptides in Mare Milk Under Different Heat Treatment Conditions.
Foods (Basel, Switzerland)    November 10, 2024   Volume 13, Issue 22 3592 doi: 10.3390/foods13223592
Lou X, Shao W, Wu Y, Ma H, Chen H, Zheng N, Zhao Y.Active peptides in mare milk have unique biological activities, but how the bioactive protein in mare's milk changes under the influence of temperature has not been fully studied. Therefore, in this study, the differential expression of bioactive peptides potentially present in horse milk under different heat treatment conditions was investigated for the first time using peptidomic and bioinformatic techniques. We collected a total of 15 samples. In this study, we divided the samples into five groups, specifically, 65 °C, 30 min; 72 °C, 15 min; 83 °C; 10 min; 95 °C, 5 min; and an untreated...
Is Butyrate Concentration in the Equine Gastrointestinal Tract Altered During and After Surgery for Treatment of Large Colon Obstruction?
Animals : an open access journal from MDPI    November 8, 2024   Volume 14, Issue 22 3203 doi: 10.3390/ani14223203
Barton CK, Hassel DM, Anders K, Weir TL.A major cause of morbidity and mortality in horses with large colon obstructive lesions is injury to the colonic mucosal barrier from ischemic injury. Since butyrate has been shown to play a critical role in the maintenance of a healthy mucosal barrier, it may play a role in the recovery process. This study's objective was to determine whether the differences in butyrate concentrations existed between horses with surgical large colon obstructive lesions and healthy horses both during and after surgery. Eleven horses presenting with surgical colic lesions were enrolled; colonic samples were acq...
Differences in physicochemical properties and proteomics analysis of spray- and freeze-dried milk powders from bovine, goat, and horse sources.
Journal of dairy science    November 7, 2024   S0022-0302(24)01277-3 doi: 10.3168/jds.2024-25146
Zhou S, Zhang X, Zhang J, Zang C, Fan R, Wang J, Guo T, Han R, Yang Y.Milk powder, a nutrient-rich dairy product, lacks comprehensive information summarizing its specific properties when produced by spray- and freeze-dried technologies from different sources. Therefore, this study investigated the differences in physicochemical properties, microstructure, and proteome of spray- and freeze-dried milk powders from bovine, goat, and horse sources. The results revealed that spray-dried milk powder exhibited a smaller particle size, lower air content within the powder particles, inferior reconstitution properties, and lower lactose crystallinity compared with freeze-...
Analysis of stallion spermatozoa metabolism using Agilent Seahorse XFp Technology.
Animal reproduction science    November 3, 2024   Volume 271 107633 doi: 10.1016/j.anireprosci.2024.107633
Ortiz-Rodriguez JM, Bucci D, Tovar-Pascual L, Granata S, Spinaci M, Nesci S.Sperm metabolism consists of a sophisticated network of biochemical reactions and varies between species, resulting in different metabolic strategies for ATP production to maintain sperm functionality. ATP can be produced through glycolysis or in the mitochondria by oxidative phosphorylation (OXPHOS). Since OXPHOS is the predominant metabolic pathway in horses spermatozoa, various assessments of mitochondrial activity are used to evaluate fertility, utilizing techniques such as fluorescent probes analysed via microscopy or flow cytometry, and polarographic electrode assays to measure current f...
Age-dependent haemogram and sex-dependent serum biochemistry values in semi-feral Konik horses.
Schweizer Archiv fur Tierheilkunde    October 29, 2024   Volume 166, Issue 11 582-592 doi: 10.17236/sat00437
Thielebein J, Bartling B, Hönicke C, Schmicke M.Semi-feral, free-roaming Konik polski horses are used in some European countries for preserving semi-open pasture landscapes. The estimation of their health status is still limited by insufficient data on various blood parameters. Therefore, our study aimed at the sex- and age-dependent analysis of haemogram and selected biochemistry parameters in healthy, semi-feral Koniks. In order to reach this aim, we took blood samples from 53 female and 18 male (8 uncastrated, 10 castrated) Koniks living in two Middle German nature reserves. They were of different age (9-266 months) and without signs of ...
Calcium propionate is an alkalizing agent in exercising Standardbreds.
Journal of equine veterinary science    October 29, 2024   Volume 143 105212 doi: 10.1016/j.jevs.2024.105212
Herbst AC, Puglia AT, Malinowski K, McKeever KH.The objective of this study was to determine if calcium propionate is an alkalizing agent in exercising Standardbreds and if it alters plasma glucose and serum insulin concentrations. This study used a randomized crossover design to test the hypotheses that calcium propionate alters total CO2 (tCO), Ca, pH, strong ion difference (SID), glucose, and insulin in Standardbreds completing a simulated race test (SRT) on a high-speed equine treadmill. Blood was collected from eight horses (mean age ± SD = 16 ± 2.7 years; range = 13 - 21 years) at 10 and 5 min prior to treatment or control administr...
Preparation and tissue structure analysis of horse bone collagen peptide.
Scientific reports    October 28, 2024   Volume 14, Issue 1 25687 doi: 10.1038/s41598-024-75960-7
Wu J, Na H, Bai F, Li S, Gao H, Sha R.Horse bone is rich in collagen, with a composition similar to that of human collagen. Collagen peptides supply nutrients needed for human growth that act as antioxidants, lower blood pressure. This study explored the extraction of collagen and the preparation of collagen short peptides from Mongolian horse bones. Bones were collected from horses of varying ages, and the collagen content along with calcium salt distribution were observed through staining and imaging analyses. Next, the bones were processed into a powder and then subjected to ultra-high-pressure processing for degreasing. The de...
Effect of furosemide on comprehensive renin-angiotensin-aldosterone system activity of Thoroughbred horses.
Journal of veterinary internal medicine    October 22, 2024   doi: 10.1111/jvim.17208
Lehman ML, Domenig O, Ames MK, Morgan JM.Furosemide, a commonly used diuretic, activates the renin-angiotensin-aldosterone system (RAAS) in other species. Little is known about RAAS peptide activation in horses. Objective: To evaluate equilibrium analysis as a practical method for RAAS quantification in horses and describe the RAAS response to a single dose of furosemide. We hypothesize that furosemide would cause transient increase in RAAS peptides in horses. Methods: 14 healthy adult thoroughbreds from a university teaching herd. Methods: Horses received either furosemide (1 mg/kg IV) or saline IV in a crossover study design. Pro...
Sex hormone-binding globulin promotes the osteogenic differentiation potential of equine adipose-derived stromal cells by activating the BMP signaling pathway.
Frontiers in endocrinology    October 17, 2024   Volume 15 1424873 doi: 10.3389/fendo.2024.1424873
Irwin-Huston JM, Bourebaba L, Bourebaba N, Tomal A, Marycz K.Musculoskeletal injuries and chronic degenerative diseases pose significant challenges in equine health, impacting performance and overall well-being. Sex Hormone-Binding Globulin (SHBG) is a glycoprotein determining the bioavailability of sex hormones in the bloodstream, and exerting critical metabolic functions, thus impacting the homeostasis of many tissues including the bone. Unassigned: In this study, we investigated the potential role of SHBG in promoting osteogenesis and its underlying mechanisms in a model of equine adipose-derived stromal cells (ASCs). An SHBG-knocked down model has b...
Plasma Lipidomics Reveals Lipid Signatures of Early Pregnancy in Mares.
International journal of molecular sciences    October 15, 2024   Volume 25, Issue 20 11073 doi: 10.3390/ijms252011073
Perera TRW, Bromfield EG, Gibb Z, Nixon B, Sheridan AR, Rupasinghe T, Skerrett-Byrne DA, Swegen A.Understanding the systemic biochemistry of early pregnancy in the mare is essential for developing new diagnostics and identifying causes for pregnancy loss. This study aimed to elucidate the dynamic lipidomic changes occurring during the initial stages of equine pregnancy, with a specific focus on days 7 and 14 post-ovulation. By analysing and comparing the plasma lipid profiles of pregnant and non-pregnant mares, the objective of this study was to identify potential biomarkers for pregnancy and gain insights into the biochemical adaptations essential for supporting maternal recognition of pr...
Assessing muscle function and oxidative profile in criollo horses participating in long loop rodeo before and after exercise.
Brazilian journal of veterinary medicine    October 11, 2024   Volume 46 e005823 doi: 10.29374/2527-2179.bjvm005823
Pereira AT, Pozzobon R, Dos Anjos BL, Chaves LT, Smilgys ÉC, Thoma AJS, Lunkes VL, de Oliveira JS, de Andrade CM, Soares JCM.Criollo breed is quite popular in Brazil, mainly in the Southern Region, and the long loop rodeo is one of the main sports modality adapted to this breed. The purpose of this study was to assess changes in the muscle and oxidative functions caused by exercises practiced in this sports modality. Data such as exercising frequency, covered distance, mean duration time and number of runs, and blood samples at the following times: before the tournament, on the final day of the event, and 18-24 h after the end of the tournament were collected. Were assessed lactate and the enzymes aspartate aminotra...
Featuring long-loop tournaments: breeding and training profiles and blood changes in criollo horses before and after exercise.
Brazilian journal of veterinary medicine    October 11, 2024   Volume 46 e005723 doi: 10.29374/2527-2179.bjvm005723
Pereira AT, Pozzobon R, Dos Anjos BL, Kunz AR, Chaves LT, Smilgys EC, Andrade BDS, Lunkes VL, de Andrade CM, Costa MM.Long-loop rodeo is a major competition for Criollo horses. We aimed to feature long-loop tournaments and to assess the profiles of competing horses. The animals (n = 49) were registered, and their body mass/scores, information about breeding, feeding, and training management, loaded weight, and tournament track (n = 11) were collected; speeds reached were estimated. Heart and respiratory rates of horses (n = 27) were collected before the tournament, on the final day of the event, and 18-24 h after the end of the tournament. Blood samples were collected from the animals at the same time. Blood ...
Evaluation of Two Commercial ELISA Kits for Measuring Equine Serum Gastrin Compared to Radioimmunoassay.
Animals : an open access journal from MDPI    October 11, 2024   Volume 14, Issue 20 2937 doi: 10.3390/ani14202937
Vokes JR, Gedye KR, Lovett AL, de Kantzow MC, Shan R, Steel CM, Sykes BW.Gastrin is an important hormone involved in gastric acid secretion. Despite its importance, validated methods other than radioimmunoassay (RIA) to assess serum gastrin concentrations in horses are lacking. This study aims to determine the agreement between ELISA and RIA in quantifying equine serum gastrin concentrations. Serum gastrin concentrations were quantified using two ELISA kits and RIA. Samples (196) from 14 horses at different time points were analyzed using one ELISA kit and RIA, selected samples (7) were analyzed using a second ELISA kit, and the correlation between methods was calc...
Accuracy and validation of a point-of-care blood glucose monitoring system for use in horses.
Frontiers in veterinary science    October 10, 2024   Volume 11 1436714 doi: 10.3389/fvets.2024.1436714
Velineni S, Schiltz P, Chang KH, Peng YM, Cowles B.Abnormal blood glucose (BG) levels often seen in critically ill horses are significantly associated with adverse patient outcomes and increased mortality. Rapid and accurate BG monitoring is now considered an essential component of evidence-based equine practice and can provide critical information quickly for treatment. Although several point-of-care (POC) BG monitoring hand-held devices are commercially available for veterinary use, none contains a unique algorithm validated for use in horses. The AlphaTrak 3 (AT3) BG monitoring system is a first-of-its-kind device with an equine-specific al...
Protein profile of extracellular vesicles derived from adult Parascaris spp.
Parasites & vectors    October 10, 2024   Volume 17, Issue 1 426 doi: 10.1186/s13071-024-06502-3
Manikantan V, Ripley NE, Nielsen MK, Dangoudoubiyam S.Parascaris spp. represent a significant threat to equine health worldwide, particularly in foals. The long-term survival of parasites in the host necessitates persistent modulation of the host immune response. Intercellular communication achieved through the exchange of molecules via extracellular vesicles (EVs) released from the parasite could be a crucial factor in this regard. This study aimed to isolate and characterize EVs released by adult male and female Parascaris worms and conduct a proteomic analysis to identify sex-specific proteins and potential immunomodulatory factors. Methods: L...
Biochar captures ammonium and nitrate in easily extractable and strongly retained form without stimulating greenhouse gas emissions during composting.
Journal of environmental quality    October 9, 2024   doi: 10.1002/jeq2.20634
Busch F, Leal ODA, Siebers N, Brüggemann N.During composting of organic waste, nitrogen is lost through gaseous forms and ion leaching. Biochar has been shown to capture mineral nitrogen (N: NH and NO ) from compost, which we hypothesize reduces NO formation. However, associating N captured by biochar with the dynamics of NO and other greenhouse gas (GHG) emissions during composting remains unstudied and was the aim of this work. We composted (outdoor for 148 days) together kitchen scraps (43.3% dw, where dw is dry weight), horse manure (40.9% dw), and wheat (Triticum aestivum L) straw (15.8% dw) without (Control) or with biochar (Bc,...
Physiological and metabolic effects of short-term dopamine reduction in healthy horses using a tyrosine hydroxylase inhibitor (alpha-methyl-para-tyrosine).
Domestic animal endocrinology    October 5, 2024   Volume 90 106891 doi: 10.1016/j.domaniend.2024.106891
Galinelli NC, Bamford NJ, Erdody ML, Warnken T, de Laat MA, Sillence MN, Harris PA, Bailey SR.Alpha-methyl-para-tyrosine (AMPT) is a reversible inhibitor of tyrosine hydroxylase, the rate-limiting enzyme in catecholamine synthesis. This study aimed to determine whether AMPT could reduce dopamine concentrations in horses. Six healthy adult Standardbred geldings were administered AMPT (40 mg/kg BW, orally) or placebo in a randomised crossover study design. Clinical examination findings were recorded, and blood samples were collected for up to 6 h after administration of AMPT or placebo, for measurement of blood glucose, plasma ACTH and cortisol concentrations, and plasma metabolomic anal...
Overexpression of the key metabolic protein Carnitine Palmitoyl Transferase 1A (CPT1A) in equine sarcoid.
Journal of equine veterinary science    October 2, 2024   Volume 143 105205 doi: 10.1016/j.jevs.2024.105205
Martano M, Power K, Cuccaro B, Razzuoli E, Maiolino P, Restucci B.The equine sarcoid is the most common skin neoplasia of fibroblastic origin in horses, characterized by an excessive accumulation of extracellular matrix produced by sarcoid fibroblasts under hypoxic condition. Neoplastic cells can adapt to hypoxia by using alternative energy sources, particularly those that arise from fatty acid oxidation (FAO). The Carnitine Palmitoyl Transferase 1A (CPT1A) belongs to Carnitine System (CS) and promotes the entrance of fatty acids into the mitochondria for β-oxidation. In this study, CPT1A expression was comparatively addressed in 25 equine sarcoids and 5 no...
Investigating the Suitability of Mare’s Milk-Derived Exosomes as Potential Drug Carriers.
Biomolecules    October 1, 2024   Volume 14, Issue 10 1247 doi: 10.3390/biom14101247
Sergazy S, Zhetkenev S, Shulgau Z, Chulenbayeva L, Kamyshanskiy Y, Nurgaziyev M, Nurgozhina A, Mukhanbetzhanova Z, Berikkhanova K, Gulyayev A....Exosomes are cell-derived, membrane-surrounded particles that deliver bioactive molecules to various cells. Due to their small size, low immunogenicity, extended blood circulation, and involvement in cellular communication, they hold potential as effective drug carriers. Exosomes are present in various biological fluids, including mare's milk, a traditional drink in Central Asia. This study aims to compare exosome isolation methodologies and determine the stability of mare's milk-derived exosomes as potential therapeutic carriers. Three extraction methods-immunoprecipitation, size exclusion ch...
A multi-task learning model for clinically interpretable sesamoiditis grading.
Computers in biology and medicine    September 25, 2024   Volume 182 109179 doi: 10.1016/j.compbiomed.2024.109179
Guo L, Tahir AM, Hore M, Collins A, Rideout A, Wang ZJ.Sesamoiditis is a common equine disease with varying severity, leading to increased injury risks and performance degradation in horses. Accurate grading of sesamoiditis is crucial for effective treatment. Although deep learning-based approaches for grading sesamoiditis show promise, they remain underexplored and often lack clinical interpretability. To address this issue, we propose a novel, clinically interpretable multi-task learning model that integrates clinical knowledge with machine learning. The proposed model employs a dual-branch decoder to simultaneously perform sesamoiditis grading ...
Equine metabolic investigation of the phosphodiesterase-4 inhibitor ibudilast as a potential performance enhancer.
Rapid communications in mass spectrometry : RCM    September 23, 2024   Volume 38, Issue 23 e9916 doi: 10.1002/rcm.9916
Philip M, Kal AKK, Subhahar MB, Karatt TK, Graiban FM, Ajeebsanu MM, Joseph M, Jose SV.Phosphodiesterase 4 (PDE4) inhibitors are a newer class of drugs that induce bronchodilation and have anti-inflammatory effects, making them susceptible to misuse as performance enhancers in competitive sports. Methods: This study explores the metabolic conversion of PDE4 inhibitor ibudilast in thoroughbred horses after oral administration and in vitro using equine liver microsomes and Cunninghamella elegans. A liquid chromatography-high resolution mass spectrometry method was used to postulate the plausible structures of the detected metabolites. Results: A total of 20 in vivo metabolites wer...
Extraction and Physicochemical Characterization of Hydroxyapatites From Horse Humerus Bones of Different Ages (1, 3, 6, and 8 Years old) Calcined at Low Temperature.
Journal of biomedical materials research. Part B, Applied biomaterials    September 19, 2024   Volume 112, Issue 10 e35484 doi: 10.1002/jbm.b.35484
Correa-Piña BA, Castillo-Paz AM, Davila U, Rodriguez-Garcia ME.The aim of this work is to investigate the changes in the physicochemical properties of hydroxyapatite (HAp) extracted from horse humerus bones of different ages (1, 3, 6, and 8 years) subjected to low temperature calcination (600°C). Thermal analysis revealed significant mass loss due to water, collagen, organic compounds, carbonates, and age-related magnesium out-diffusion. Higher fat content in older bones contributed to increased mass loss. Phosphorus content remained constant across age groups, while calcium and sodium showed age-related fluctuations. Magnesium levels decreased with ag...
Effect of protein acetylation on capacitation of stallion sperm.
Journal of equine veterinary science    September 18, 2024   Volume 142 105195 doi: 10.1016/j.jevs.2024.105195
Aguiar LH, Pinto CRF.Sperm capacitation is considered the main factor limiting conventional in vitro fertilization (IVF) in horses. A recent scientific breakthrough in sperm processing for IVF in horses has resulted in embryos and foals being produced; however, various aspects of the IVF process remain to be fully elucidated. Lysine acetylation has been shown to play a role in sperm capacitation in several species and the objective of this study was to detect and evaluate this process in the horse. Ejaculates of two stallions were collected and incubated in different conditions with deacetylase inhibitors to induc...
Effects of maternal dietary supplementation with antioxidants on clinical status of mares and their foal.
BMC veterinary research    September 11, 2024   Volume 20, Issue 1 404 doi: 10.1186/s12917-024-04252-z
Del Prete C, Vastolo A, Pasolini MP, Cocchia N, Montano C, Cutrignelli MI.The peripartum period constitutes a delicate physiological moment in mares showing a transient state of oxidative stress. Diet supplementation with antioxidants during pregnancy in women appears to have a beneficial effect on mother and neonate health. The aim of this work was to evaluate the effects of diet supplementation with a commercial product containing a mix of antioxidants (Oxyliver®, Candioli) on the length of gestation, weight, and haemato-biochemical parameters in Italian Salernitano mares and their newborn foals. Eight late-term pregnant mares were randomly divided into two group...
Bioreactance noninvasive cardiac output monitoring in euvolemic adult horses subjected with 7.2% saline and 6% hydroxyethyl starch (130/0.4) solution infusions.
Journal of veterinary emergency and critical care (San Antonio, Tex. : 2001)    September 10, 2024   doi: 10.1111/vec.13418
Long AE, Hurcombe S, Hopster K.To compare the ability of bioreactance noninvasive cardiac output (BR-NICO) with thermodilution cardiac output (TDCO) for the measurement of cardiac output (CO) in healthy adult horses receiving 2 different IV volume replacement solutions. Methods: Prospective randomized crossover study from September to November 2021. Methods: University teaching hospital. Methods: Six university-owned adult horses. Methods: Horses were randomly assigned to receive 7.2% hypertonic saline solution (HSS) or 6% hydroxyethyl starch (130/0.4) solution (HETA). BR-NICO and TDCO were measured simultaneously at baseli...
The Effect of Different Storage Temperatures over Time on the pH of Mammary Gland Secretions in Periparturient Mares.
Animals : an open access journal from MDPI    September 6, 2024   Volume 14, Issue 17 doi: 10.3390/ani14172598
Canisso IF, Amorim GBAG, Magalhaes HB.The objectives of this study were (i) to determine pH and electrolyte concentrations in MGS collected prepartum and at parturition, (ii) to characterize mare milk pH during the first week postpartum, and (iii) to evaluate pre-foaling MGS pH at three storage temperatures. This study outlined two hypotheses: (i) all mares exhibit acidic pH, increased calcium, magnesium, and potassium, and reduced sodium concentrations regardless of prepartum pH and electrolytes; (ii) pre-foaling MGS pH varies with storage temperature and time in an initial value-dependent manner. Twenty-three multiparous mares w...
Effects of storage-handling methods on nutrient analysis of fresh-forage samples.
Journal of equine veterinary science    September 4, 2024   Volume 142 105182 doi: 10.1016/j.jevs.2024.105182
Kerley BS, Harris P, Lea K, McClendon M, Adams AA.Forage low in nonstructural carbohydrates (NSC) is recommended for insulin dysregulated (ID) horses, indicating the importance of an accurate forage analysis. However, it remains to be fully understood how handling forage samples pre-analysis impacts nutrient values. The objective of this study was to compare the effect of fresh-forage storage-handling methods: microwave-oven (MO; 9kw; 70s then stored at -20℃), room temperature (20℃), 3℃, -20℃, and -80℃. Subsamples collected from a mixed-grass pasture and a ryegrass plot were analyzed for crude protein (CP), water soluble carbohydrat...