The Latin Square Design is a statistical method used in experimental research to control for variability across two factors, such as time and treatment, in equine studies. This design arranges treatments in a square grid, ensuring that each treatment appears exactly once in each row and column. In equine research, the Latin Square Design is employed to minimize the effects of confounding variables, allowing for more accurate assessment of treatment effects. By systematically organizing variables, researchers can isolate the impact of specific interventions on horses. This section includes peer-reviewed research studies and scholarly articles focusing on the application, methodology, and outcomes of using the Latin Square Design in equine research.
Köninger M, von Velsen-Zerweck A, Eiberger C, Löffler C, Töpper A, Visscher C, Reckels B, Vervuert I.The aim of this study was to examine the nutrient composition and feed hygiene of alfalfa as well as investigate the feed intake, blood, urine and faecal parameters of horses fed alfalfa haylage (AS) compared with alfalfa hay (AH) and meadow hay (MH). A total of 11 geldings were fed ad libitum (2.1% dry matter (DM) of body weight (BW)) with alfalfa haylage, alfalfa hay and meadow hay (MH) in a Latin square design. On days 0 and 21 of the feeding period, blood samples were analysed for kidney and liver parameters. Faecal samples were analysed for pH, DM and short-chain fatty acids (SCFAs). Spon...
White JM, Colbath AC, Schott HC.Treatment with phenylbutazone (nonselective COX inhibitor) decreases the diuretic and natriuretic effects of furosemide by nearly 30% but the effects of COX-2 specific inhibitors (firocoxib) and atypical NSAIDs (dipyrone) are unknown. Objective: Furosemide-induced diuresis after pretreatment with firocoxib or dipyrone is diminished to a lesser extent than after pretreatment with phenylbutazone. Methods: Eight healthy mares. Methods: Each mare received 4 treatments in a prospective experimental crossover study using a replicated 4 × 4 Latin Square design: furosemide alone (FU), furosemide an...
Springer RW, Mason AC, Cross TD, Guay KA, Raub RH, Wellmann KB, Jones TN.With greater availability of hemp (Cannabis sativa L.) due to recent legislation, hempseed meal (HSM) should be evaluated as a potential feedstuff for livestock. The study objective was to evaluate the palatability of HSM in horses. Trial 1 used a modified 3 × 3 Latin Square design utilizing three geldings and three mares. Feeding periods (6d) consisted of two feedings/d with a washout phase (3d) before each treatment phase (3d). During treatment, horses were offered soybean meal pellets (SBM), rice bran pellets (RBP), or beet pulp pellets (BPP) in one bucket (500 g) and HSM in a second buc...
Arana Valencia N, Thompson DL, Southerland CV.Four experiments assessed factors affecting prolactin responses to sulpiride administration in horses. Experiment 1 compared the efficacy of the (-) enantiomer of sulpiride to that of the commonly used (+/-) racemic mixture. Mares were used in an 8 × 8 Latin square to compare the prolactin responses to four doses of levosulpiride to four corresponding doses of the racemic mixture at twice the dose. Responses at each dose indicated equal and similar (P > .1) responses. Experiment 2 compared the efficacy of 1 gram of orally administered racemic sulpiride to 100 mg of intramuscularly injec...
Hussein H, Ishihara A, Menendez M, Bertone A.Plasma pharmacokinetic (PK) and bone resorption biomarker [carboxy-terminal cross-linking telopeptide of type I collagen (CTX-1)] analyses were performed following single and multiple oral dose protocols of a Cathepsin K inhibitor (VEL-0230) in horses. Outcomes included plasma and urine drug and CTX-1 concentrations. In the dose range study, 2, 4, and 8 mg/kg body weight (b.w.) doses were administered in a Latin square design to three mares and evaluated for 1 week. Based on the PK characteristics of VEL-0230, 4 mg/kg b.w. was selected for the dose interval study in which 3.25 days (d) and 7 d...
Fernandes CB, Devito LG, Martins LR, Blanco ID, de Lima Neto JF, Tsuribe PM, Gonçalves CG, da Cruz Landim-Alvarenga F.Knowledge on parthenogenetic activation of oocytes is important to improve the efficiency of nuclear transfer (NT) and intracytoplasmic sperm injection (ICSI) because artificial activation of oocyte (AOA) is an essential step to achieve embryo production. Although different procedures for AOA have been established, the efficiency of in vitro production of embryos remains low, especially in equines and Bos taurus bovines. In an attempt to improve the techniques of NT and ICSI in bovine and equine species, we tested different combinations of drugs that had different mechanisms of action for the ...
Brøkner C, Austbø D, Næsset JA, Knudsen KE, Tauson AH.The working hypothesis was that a minor postprandial caecal pH decline would affect apparent total tract digestibility (ATTD) of the fibre fraction in horses and, hence, that soluble fibre would amplify fermentation and consequently increase ATTD of fibre. This study was a 4 × 4 Latin Square design with a sequence of 17 days adaptation to the ration followed by 8 sampling days. The feed rations consisted of only timothy hay (Group H), hay plus molassed sugar beet pulp combined with either whole oats (Group OB) or barley (Group BB) and hay plus loose chaff based concentrate (Group M). Four hor...
Rosenmeier JG, Strathe AB, Andersen PH.To measure coronary band temperature (CBT) in healthy horses fed high-fructan or low-carbohydrate diets and to analyze the association of CBT with diet, time of day, and ambient temperature. Methods: 6 healthy horses. Methods: Horses were fed 3 diets (treatment 1, 1 g of fructan/kg fed daily in the morning; treatment 2, 1 g of fructan/kg fed daily in the afternoon; and treatment 3, a low-carbohydrate [7.2%] diet) in a 3 × 3 Latin square study design. For each horse, the CBT of all 4 limbs as well as rectal and ambient temperatures were recorded by use of infrared thermometry and standard ther...
de Fombelle A, Veiga L, Drogoul C, Julliand V.This trial was conducted to determine the extent of prececal starch digestibility depending on the botanical origin of starch and on diet characteristics (i.e., composition and feeding pattern). The prececal disappearance of six substrates (oats, barley, corn, horse bean, potato, and wheat) was measured in four cannulated horses fed (as-fed basis) 11.8 g/kg BW of a high-fiber (HF) or high-starch (HS) pelleted feed and 10.0 g/kg BW of meadow hay using the mobile bag technique (MBT). The daily feeding pattern was either three meals (two meals of pellets and one meal of hay) or five meals (three ...
Frank N, Sojka JE, Latour MA.Blood lipid and lipoprotein concentrations were measured and compared between euthyroid and thyroidectomized mares on low-fat or high-fat diets to test the hypothesis that hypothyroidism alters the blood lipid response to higher dietary fat intake. Four healthy adult mares and four adult mares that had been thyroidectomized 3 to 6 mo earlier were placed on low-fat or high-fat diets according to a replicated 2 x 2 Latin square design consisting of two 5-wk feeding periods separated by a 2-wk washout interval. Plasma lipid concentrations were measured at 0, 3, 4, and 5 wk, and plasma lipase acti...
Cadario ME, Merritt AM, Archbald LF, Thatcher WW, LeBlanc MM.Intrauterine pressure was measured in 4 reproductively normal mares and 4 mares with delay in uterine clearance after administration of oxytocin to determine if intrauterine pressure varied between dosage and group. Changes in intrauterine pressure were measured during estrus, when a follicle was > or =35 mm, using a Millar "Mikro-tip" catheter that had 3 discrete pressure sensors/channels. Mares received 4 different treatments of 10, 5, 2.5 or 0 IU (vehicle) of oxytocin. The protocol for each treatment consisted of a 10-min baseline recording, administration of treatment and measurement of...
England GC, Clarke KW, Goossens L.The sedative effects of a new alpha 2-adrenoceptor agonist, romifidine, were compared with those of xylazine and detomidine. Five horses were treated with two doses of romifidine (40 micrograms/kg body weight and 80 micrograms/kg body weight), two doses of detomidine (10 micrograms/kg body weight and 20 micrograms/kg body weight) and one dose of xylazine (1 mg/kg body weight) given by intravenous injection using a Latin-square design. The dose of 80 micrograms/kg romifidine appeared equipotent to 1 mg/kg xylazine and 20 micrograms/kg detomidine, although at these doses both xylazine and detomi...
Glade MJ.Mature geldings at maintenance were fed different diets in a 4 x 4 Latin square design balanced to account for residual effects in an attempt to determine whether differences in the digestibility of the fibrous portions of feedstuffs would influence dietary nitrogen (N) requirements. Diet 1 contained corn and soybean meal (SBM); diet 2, corn, corn oil and urea; diet 3, corn, SBM, straw and urea; diet 4, corn, alfalfa and urea. Urea supplied 50% of the total N in diets 2 and 3 and 39% of the total N in diet 4. The diets were fed in amounts that met National Research Council (NRC) recommendation...
England GC, Clarke KW, Goossens L.The sedative effects of a new alpha 2-adrenoceptor agonist, romifidine, were compared with those of xylazine and detomidine. Five horses were treated with two doses of romifidine (40 micrograms/kg body weight and 80 micrograms/kg body weight), two doses of detomidine (10 micrograms/kg body weight and 20 micrograms/kg body weight) and one dose of xylazine (1 mg/kg body weight) given by intravenous injection using a Latin-square design. The dose of 80 micrograms/kg romifidine appeared equipotent to 1 mg/kg xylazine and 20 micrograms/kg detomidine, although at these doses both xylazine and detomi...
de Fombelle A, Veiga L, Drogoul C, Julliand V.This trial was conducted to determine the extent of prececal starch digestibility depending on the botanical origin of starch and on diet characteristics (i.e., composition and feeding pattern). The prececal disappearance of six substrates (oats, barley, corn, horse bean, potato, and wheat) was measured in four cannulated horses fed (as-fed basis) 11.8 g/kg BW of a high-fiber (HF) or high-starch (HS) pelleted feed and 10.0 g/kg BW of meadow hay using the mobile bag technique (MBT). The daily feeding pattern was either three meals (two meals of pellets and one meal of hay) or five meals (three ...
Brøkner C, Austbø D, Næsset JA, Knudsen KE, Tauson AH.The working hypothesis was that a minor postprandial caecal pH decline would affect apparent total tract digestibility (ATTD) of the fibre fraction in horses and, hence, that soluble fibre would amplify fermentation and consequently increase ATTD of fibre. This study was a 4 × 4 Latin Square design with a sequence of 17 days adaptation to the ration followed by 8 sampling days. The feed rations consisted of only timothy hay (Group H), hay plus molassed sugar beet pulp combined with either whole oats (Group OB) or barley (Group BB) and hay plus loose chaff based concentrate (Group M). Four hor...
Cadario ME, Merritt AM, Archbald LF, Thatcher WW, LeBlanc MM.Intrauterine pressure was measured in 4 reproductively normal mares and 4 mares with delay in uterine clearance after administration of oxytocin to determine if intrauterine pressure varied between dosage and group. Changes in intrauterine pressure were measured during estrus, when a follicle was > or =35 mm, using a Millar "Mikro-tip" catheter that had 3 discrete pressure sensors/channels. Mares received 4 different treatments of 10, 5, 2.5 or 0 IU (vehicle) of oxytocin. The protocol for each treatment consisted of a 10-min baseline recording, administration of treatment and measurement of...
Frank N, Sojka JE, Latour MA.Blood lipid and lipoprotein concentrations were measured and compared between euthyroid and thyroidectomized mares on low-fat or high-fat diets to test the hypothesis that hypothyroidism alters the blood lipid response to higher dietary fat intake. Four healthy adult mares and four adult mares that had been thyroidectomized 3 to 6 mo earlier were placed on low-fat or high-fat diets according to a replicated 2 x 2 Latin square design consisting of two 5-wk feeding periods separated by a 2-wk washout interval. Plasma lipid concentrations were measured at 0, 3, 4, and 5 wk, and plasma lipase acti...
Hussein H, Ishihara A, Menendez M, Bertone A.Plasma pharmacokinetic (PK) and bone resorption biomarker [carboxy-terminal cross-linking telopeptide of type I collagen (CTX-1)] analyses were performed following single and multiple oral dose protocols of a Cathepsin K inhibitor (VEL-0230) in horses. Outcomes included plasma and urine drug and CTX-1 concentrations. In the dose range study, 2, 4, and 8 mg/kg body weight (b.w.) doses were administered in a Latin square design to three mares and evaluated for 1 week. Based on the PK characteristics of VEL-0230, 4 mg/kg b.w. was selected for the dose interval study in which 3.25 days (d) and 7 d...
Glade MJ.Mature geldings at maintenance were fed different diets in a 4 x 4 Latin square design balanced to account for residual effects in an attempt to determine whether differences in the digestibility of the fibrous portions of feedstuffs would influence dietary nitrogen (N) requirements. Diet 1 contained corn and soybean meal (SBM); diet 2, corn, corn oil and urea; diet 3, corn, SBM, straw and urea; diet 4, corn, alfalfa and urea. Urea supplied 50% of the total N in diets 2 and 3 and 39% of the total N in diet 4. The diets were fed in amounts that met National Research Council (NRC) recommendation...
Fernandes CB, Devito LG, Martins LR, Blanco ID, de Lima Neto JF, Tsuribe PM, Gonçalves CG, da Cruz Landim-Alvarenga F.Knowledge on parthenogenetic activation of oocytes is important to improve the efficiency of nuclear transfer (NT) and intracytoplasmic sperm injection (ICSI) because artificial activation of oocyte (AOA) is an essential step to achieve embryo production. Although different procedures for AOA have been established, the efficiency of in vitro production of embryos remains low, especially in equines and Bos taurus bovines. In an attempt to improve the techniques of NT and ICSI in bovine and equine species, we tested different combinations of drugs that had different mechanisms of action for the ...
Rosenmeier JG, Strathe AB, Andersen PH.To measure coronary band temperature (CBT) in healthy horses fed high-fructan or low-carbohydrate diets and to analyze the association of CBT with diet, time of day, and ambient temperature. Methods: 6 healthy horses. Methods: Horses were fed 3 diets (treatment 1, 1 g of fructan/kg fed daily in the morning; treatment 2, 1 g of fructan/kg fed daily in the afternoon; and treatment 3, a low-carbohydrate [7.2%] diet) in a 3 × 3 Latin square study design. For each horse, the CBT of all 4 limbs as well as rectal and ambient temperatures were recorded by use of infrared thermometry and standard ther...
Arana Valencia N, Thompson DL, Southerland CV.Four experiments assessed factors affecting prolactin responses to sulpiride administration in horses. Experiment 1 compared the efficacy of the (-) enantiomer of sulpiride to that of the commonly used (+/-) racemic mixture. Mares were used in an 8 × 8 Latin square to compare the prolactin responses to four doses of levosulpiride to four corresponding doses of the racemic mixture at twice the dose. Responses at each dose indicated equal and similar (P > .1) responses. Experiment 2 compared the efficacy of 1 gram of orally administered racemic sulpiride to 100 mg of intramuscularly injec...
Springer RW, Mason AC, Cross TD, Guay KA, Raub RH, Wellmann KB, Jones TN.With greater availability of hemp (Cannabis sativa L.) due to recent legislation, hempseed meal (HSM) should be evaluated as a potential feedstuff for livestock. The study objective was to evaluate the palatability of HSM in horses. Trial 1 used a modified 3 × 3 Latin Square design utilizing three geldings and three mares. Feeding periods (6d) consisted of two feedings/d with a washout phase (3d) before each treatment phase (3d). During treatment, horses were offered soybean meal pellets (SBM), rice bran pellets (RBP), or beet pulp pellets (BPP) in one bucket (500 g) and HSM in a second buc...
White JM, Colbath AC, Schott HC.Treatment with phenylbutazone (nonselective COX inhibitor) decreases the diuretic and natriuretic effects of furosemide by nearly 30% but the effects of COX-2 specific inhibitors (firocoxib) and atypical NSAIDs (dipyrone) are unknown. Objective: Furosemide-induced diuresis after pretreatment with firocoxib or dipyrone is diminished to a lesser extent than after pretreatment with phenylbutazone. Methods: Eight healthy mares. Methods: Each mare received 4 treatments in a prospective experimental crossover study using a replicated 4 × 4 Latin Square design: furosemide alone (FU), furosemide an...
Köninger M, von Velsen-Zerweck A, Eiberger C, Löffler C, Töpper A, Visscher C, Reckels B, Vervuert I.The aim of this study was to examine the nutrient composition and feed hygiene of alfalfa as well as investigate the feed intake, blood, urine and faecal parameters of horses fed alfalfa haylage (AS) compared with alfalfa hay (AH) and meadow hay (MH). A total of 11 geldings were fed ad libitum (2.1% dry matter (DM) of body weight (BW)) with alfalfa haylage, alfalfa hay and meadow hay (MH) in a Latin square design. On days 0 and 21 of the feeding period, blood samples were analysed for kidney and liver parameters. Faecal samples were analysed for pH, DM and short-chain fatty acids (SCFAs). Spon...
