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Topic:Lactate

Lactate is a byproduct of anaerobic metabolism, produced when glucose is broken down for energy in the absence of sufficient oxygen. In horses, lactate levels can increase during intense physical exertion, such as racing or other high-performance activities. Elevated lactate concentrations are often associated with muscle fatigue and can provide insights into a horse's exercise physiology and conditioning status. Monitoring lactate levels can aid in assessing the horse's fitness, recovery, and adaptation to training. This page compiles peer-reviewed research studies and scholarly articles that explore the production, regulation, and implications of lactate in equine exercise physiology and performance.
Association between dynamic upper airway obstructions and fitness parameters in Standardbred racehorses during high-speed treadmill exercise.
Journal of the American Veterinary Medical Association    May 27, 2022   Volume 260, Issue 11 1343-1350 doi: 10.2460/javma.22.02.0072
Lo Feudo CM, Stucchi L, Cavicchioli P, Stancari G, Conturba B, Zucca E, Ferrucci F.To evaluate the association between the severity of dynamic upper airway obstruction (DUAO) and selected fitness parameters in Standardbred racehorses measured by treadmill test. 191 Standardbred racehorses that underwent a standardized diagnostic protocol for poor performance evaluation between 2002 and 2021 at the Veterinary Teaching Hospital, University of Milan. Horses underwent a diagnostic protocol including clinical examination, lameness evaluation, laboratory analyses, ECG, resting upper airway endoscopy, metabolic incremental treadmill test, high-speed treadmill endoscopy, postexercis...
Evaluation of lung ventilation distribution using electrical impedance tomography in standing sedated horses with capnoperitoneum.
Veterinary anaesthesia and analgesia    April 21, 2022   Volume 49, Issue 4 382-389 doi: 10.1016/j.vaa.2022.04.004
Beazley SG, Focken A, Fernandez-Parra R, Thomas KL, Adler A, Duke-Novakovski T.To determine changes in distribution of lung ventilation with increasing intra-abdominal pressure (IAP) from carbon dioxide (CO) insufflation in standing sedated horses. Methods: Prospective experimental study. Methods: A group of six healthy adult horses. Methods: Each horse was sedated with acepromazine, detomidine and butorphanol and sedation maintained with a detomidine infusion. The horse was restrained in a stocks system and a 32 electrode electrical impedance tomography (EIT) belt was wrapped around the thorax at the fifth-sixth intercostal space. EIT images and arterial blood samples f...
Fluctuations of Physiological Variables during Conditioning of Lipizzan Fillies before Starting under Saddle.
Animals : an open access journal from MDPI    March 25, 2022   Volume 12, Issue 7 doi: 10.3390/ani12070836
Čebulj-Kadunc N, Frangež R, Kruljc P.Scientific studies on the physiological responses of young horses to workloads are limited. Therefore, the aim of our study was to determine the basal values of some cardiovascular, thermoregulatory, hematological, and biochemical parameters in 10 purebred Lipizzan fillies aged 4 years in the initial phase of training, and their responses to a graded workload, i.e., by lunging for 15 min in four exercise tests at 2-week intervals. The basal values of the measured parameters were within a range for warm-blooded horses and mostly increased after exercise in all four exercise tests. Resting heart...
Profiling the Aerobic Window of Horses in Response to Training by Means of a Modified Lactate Minimum Speed Test: Flatten the Curve.
Frontiers in physiology    March 22, 2022   Volume 13 792052 doi: 10.3389/fphys.2022.792052
De Maré L, Boshuizen B, Vidal Moreno de Vega C, de Meeûs C, Plancke L, Gansemans Y, Van Nieuwerburgh F, Deforce D, de Oliveira JE, Hosotani G....There is a great need for objective external training load prescription and performance capacity evaluation in equestrian disciplines. Therefore, reliable standardised exercise tests (SETs) are needed. Classic SETs require maximum intensities with associated risks to deduce training loads from pre-described cut-off values. The lactate minimum speed (LMS) test could be a valuable alternative. Our aim was to compare new performance parameters of a modified LMS-test with those of an incremental SET, to assess the effect of training on LMS-test parameters and curve-shape, and to identify the optim...
Evaluation of an ex vivo model of the blood-perfused equine uterus.
Theriogenology    March 9, 2022   Volume 184 82-91 doi: 10.1016/j.theriogenology.2022.02.026
Köhne M, Unruh C, Böttcher D, Tönissen A, Ulrich R, Sieme H.Uterine pathologies are the most common causes of infertility in mares. This study aimed to establish an ex vivo blood-perfused model for equine uteri and investigate the possible effects of different cycle stages (estrus, diestrus and anestrus) on the applicability of the model. Uteri (n = 13) were collected at an abattoir, flushed with preservation solution, transported to the laboratory on ice, and isolated perfused with autologous blood for 6 h (n = 12). For negative control, one uterus was handled as described but left without perfusion for 6 h. The cycle stage was determined by ex...
Stress, Metabolic and Serum Muscle-Derived Enzymes Response of Horses Employed in Wooded Area and Field Trekking Courses.
Journal of equine veterinary science    March 5, 2022   Volume 112 103919 doi: 10.1016/j.jevs.2022.103919
Arfuso F, Piccione G, Trimarchi F, Panzera MF, Giannetto C.Horse trekking is a non-competitive sport that, as other kinds of exercise, involves skeletal muscle effort and may implicate significant energy expenditure leading to stress. The aim of the current study was to assess the effect of trekking on metabolic and stress response as well as muscle damage markers in clinically healthy horses. Thirty horses were equally divided in 3 groups according to trekking course: Group A (trekking course 15 km long), Group B (trekking course 35 km long starting;), Group C (three trekking courses for three consecutive days; during the first day the trekking cours...
Effects of intravenous ethyl pyruvate on cardiopulmonary variables and quality of recovery from anesthesia in horses.
Veterinary anaesthesia and analgesia    February 7, 2022   Volume 49, Issue 3 282-290 doi: 10.1016/j.vaa.2022.01.008
Muñoz KA, Szarek M, Manfredi JM, Robertson SA, Hubbell JA, Holcombe SJ.To determine the effects of intravenous ethyl pyruvate, an anti-inflammatory with putative benefits in horses with endotoxemia, on cardiopulmonary variables during anesthesia and the quality of anesthetic recovery. Methods: Randomized, crossover, blinded experimental design. Methods: A total of six healthy Standardbred geldings, aged 13 ± 3 years and weighing 507 ± 66 kg (mean ± standard deviation). Methods: Horses were anesthetized for approximately 90 minutes on two occasions with a minimum of 2 weeks apart using xylazine for sedation, ketamine and diazepam for induction, and isoflurane i...
Interpreting abdominal fluid in colic horses: Understanding and applying peritoneal fluid evidence.
Journal of veterinary emergency and critical care (San Antonio, Tex. : 2001)    January 20, 2022   Volume 32, Issue S1 81-96 doi: 10.1111/vec.13117
Radcliffe RM, Liu SY, Cook VL, Hurcombe SDA, Divers TJ.Interpreting changes in peritoneal fluid helps clinicians manage colic and other diseases in horses. During abdominal problems in the horse, abdominal fluid characteristics such as color, turbidity, total nucleated and red blood cell counts, cytology, total protein, and l-lactate change in predictable ways, helping the clinician characterize the disease. Methods: Normal abdominal fluid in horses is odorless, clear to light yellow in color, and transparent. Peritoneal fluid becomes more turbid with increasing levels of protein, number of WBCs or RBCs, or with gross contamination following intes...
Monitoring Performance in Show Jumping Horses: Validity of Non-specific and Discipline-specific Field Exercise Tests for a Practicable Assessment of Aerobic Performance.
Frontiers in physiology    January 14, 2022   Volume 12 818381 doi: 10.3389/fphys.2021.818381
Kirsch K, Fercher C, Horstmann S, von Reitzenstein C, Augustin J, Lagershausen H.Show jumping is a highly specialized equestrian discipline that requires technical skill but also power and fitness. Monitoring the horses' aerobic performance is therefore essential in order to verify whether the training has induced the desired cardiovascular and muscular adaptations. This study therefore aimed at evaluating the validity of non-specific and discipline-specific field exercise tests for objective evaluation of aerobic performance in show jumpers. For this purpose, data obtained from horses competing at Junior and Young Rider level during show jumping competitions as well as fi...
Chemical composition and physical characteristics of faeces in horses with and without free faecal liquid – two case-control studies.
BMC veterinary research    January 3, 2022   Volume 18, Issue 1 2 doi: 10.1186/s12917-021-03096-1
Lindroth KM, Dicksved J, Vervuert I, Müller CE.Free faecal liquid (FFL) is a condition in horses characterised by two-phase (one solid and one liquid) separation of faeces. Causes of the condition are unknown, but disturbed hindgut fermentation has been suggested as it may alter biochemical composition and appearance of faeces in equines. However, information on faecal composition in horses with FFL is scarce. Faecal chemical composition (dry matter, osmolality, ash, macro minerals, short-chain fatty acids (SCFA) and pH) and physical characteristics (free liquid, sand, water holding capacity and particle size distribution) were compared in...
Effects of Standardized Exercise Tests on Plasma Thyroid Hormones’ Kinetics in Standardbred Racehorses.
Journal of equine veterinary science    December 28, 2021   Volume 110 103853 doi: 10.1016/j.jevs.2021.103853
Fazio E, Lindner A, Cravana C, Wegener J, Medica P, Hart-Mann U, Ferlazzo A.This study examined how a standard exercise test (SET) affected (1) thyroid hormones (THs) of horses and (2) the relationship between the V of horses and TH responses to the exercise in trained Standardbred racehorses (V is the velocity run at defined conditions at which a blood lactate concentration of 4 mmol/L is determined). Twelve trained Standardbred racehorses (six stallions and six mares) performed SETs until the horses' blood lactate concentration was at or above 4 mmol/L. The horses were divided into three age groups (2, 3, and 4 years old); each group consists of 4 horses respectivel...
Comparison of β-endorphin, Lactate and Cortisol Concentrations in Winning and Losing Racehorses.
Journal of equine veterinary science    December 26, 2021   Volume 110 103857 doi: 10.1016/j.jevs.2021.103857
Tavanaeimanesh H, Dashli-Boroon OJ, Corley K.Understanding the physiological and biochemical changes in racehorses can be invaluable. Accurate information in this area could result in better understanding of needs of sport horses. The aim of this study was to prove the hypothesis that biochemical changes could influence the outcome of competitions. In this study, β-endorphin was evaluated as an indicator of analgesia, lactate as an indicator of fatigue and cortisol as an indicator of stress in the first two horses and the last ones that cross the finish line. This study was performed on 44 horses participating in the 1000-meter national...
How Much Energy Vaquejada Horses Spend in a Field Simulation Test?
Animals : an open access journal from MDPI    November 30, 2021   Volume 11, Issue 12 doi: 10.3390/ani11123421
Coelho CS, Sodre TDRP, Sousa LN, Siqueira RF, Manso Filho HC, Aragona F, Fazio F.Vaquejada is a high-intensity and short-duration exercise in which helper horses (HH) are responsible to keep a bull running in a line while pull horses (PH) work to put the bull down after 100 m of running. The purpose of this study was to quantify and compare energy expenditures (EE), transport costs (COT) and metabolic energy requirements (Pmet) of horses used in Vaquejada. Thus, eight Quarter Horses, in randomly formed pairs, performed a vaquejada simulation test (VST), which consisted of three races on a sand track (130-150 m), with a 5-min interval between them. All horses used an integr...
Maternal and Neonatal Evaluation of Derived Reactive Oxygen Metabolites and Biological Antioxidant Potential in Donkey Mares and Foals.
Animals : an open access journal from MDPI    October 3, 2021   Volume 11, Issue 10 doi: 10.3390/ani11102885
Sgorbini M, Bonelli F, Percacini G, Pasquini A, Rota A.Our aim was to measure the concentrations of derived reactive oxygen metabolite (d-ROMs) and biological antioxidant potential (BAP) of donkey mares and foals at delivery and to verify the protective role of the placenta against fetal oxidative stress. A total of 15 Amiata jennies with a physiological gestation length and delivery were included together with 17 foals (two twin foalings). After delivery, maternal and foal venous blood samples were collected along with blood from the artery. Circulating lactate and plasma d-ROMs and BAP were evaluated. The Wilcoxon test for paired data was applie...
Interleukin-6 and lactate dehydrogenase expression in a novel ex vivo rocking model of equine corneal epithelial wound healing.
Veterinary ophthalmology    September 23, 2021   Volume 24, Issue 5 509-519 doi: 10.1111/vop.12935
Wehrman RF, Genschel U, Charli A, Kanthasamy AG, Allbaugh RA, Ben-Shlomo G.To establish a physiologically relevant ex vivo model of equine corneal epithelial wound healing. Methods: Fourteen equine corneas were randomly assigned to one of two groups: wounded (n = 8) or unwounded (n = 6) controls. In the wounded group, the axial corneal epithelium was removed by applying a 6 mm filter paper disk soaked in 1N-NaOH for 60 s. Corneas were subsequently cultured using an air-liquid interface model. Evaluation of corneal healing was performed daily, and culture medium was collected. Corneas were randomly assigned to undergo processing via histopathology and RNAscope i...
Equine spinal kinematics derived from different riding positions during asymmetrical bareback riding.
Journal of equine science    September 6, 2021   Volume 32, Issue 3 81-89 doi: 10.1294/jes.32.81
Nuchprayoon N, Ritruechai P, Watchararat K, Limroongruengrat W, Wongtawan T, Arya N.Hippotherapy in patients with neuromuscular dysfunction creates high focal pressure on the pony's back due to bareback riding and an asymmetrical riding position. This study aimed to investigate the acute effect of asymmetrical bareback riding on the pony's spinal kinematics, blood lactate, serum creatine kinase, heart rate, and temperament score. Eight ponies were selected, and they were walked on a treadmill for 45 min on each experimental day, including warm-up (5 min), weight-loading by mannequin (30 min), and cool-down (10 min) sessions. During the weight-loading session, three different ...
Dynamic Metabolic Response, Clotting Times and Peripheral Indices of Central Fatigue in Horse Competing in a 44 Km Endurance Race.
Journal of equine veterinary science    August 29, 2021   Volume 106 103753 doi: 10.1016/j.jevs.2021.103753
A deep knowledge on metabolic response of athlete horse to exercise is of paramount importance for the development of optimal training programs to attain equilibrium between good performance and animal welfare. This study investigated the dynamic change of metabolic, clotting times and peripheral indices of central fatigue in horses competing in an official 44 km endurance race.  From 6 horses blood samples were collected at rest (T), within 5 minutes from the end of race (T), after 30, 60 minutes after the end of race (T and T) to investigate the values of blood lactate, glucose, platelet...
Evaluation of Cardiac Arrhythmias before, during, and after Treadmill Exercise Testing in Poorly Performing Standardbred Racehorses.
Animals : an open access journal from MDPI    August 16, 2021   Volume 11, Issue 8 2413 doi: 10.3390/ani11082413
The incidence of significant arrhythmias in sport horses and knowledge about their exact influence on athletic performance need to be clarified. The aims of the present study are to report the prevalence of cardiac arrhythmias during maximal treadmill exercise in poorly performing Standardbreds, and to investigate the possible relationship of demographic, cardiac and performance indices on premature complexes (PCs). Electrocardiographic Holter recordings before, during and after treadmill exercise testing of 158 poorly performing Standardbreds were analyzed retrospectively. Fifty horses did no...
A randomised, controlled trial to determine the effect of levothyroxine on Standardbred racehorses.
Equine veterinary journal    July 28, 2021   Volume 54, Issue 3 584-591 doi: 10.1111/evj.13480
Kritchevsky J, Olave C, Tinkler S, Tropf M, Ivester K, Forsythe L, Couetil L.The use of thyroid supplement is pervasive in athletic horses although its effects on measures of performance are not known. Objective: One purpose of this study was to determine whether supra-physiologic doses of levothyroxine affect the velocities at which blood lactate was greater than 4 mmol/L (VLa4 ) and heart rate was over 150 (V150 ) and 200 (V200 ) beats per minute respectively. Additionally, a survey of post-race blood samples was also conducted to determine whether high thyroxine concentrations were common in racehorses. Methods: A randomised, crossover, trial was performed in six h...
Influence of Stretching Exercises, Warm-Up, or Cool-Down on the Physical Performance of Mangalarga Marchador Horses.
Journal of equine veterinary science    July 16, 2021   Volume 106 103714 doi: 10.1016/j.jevs.2021.103714
Farinelli F, de Rezende ASC, Fonseca MG, Lana ÂMQ, Leme FOP, Klein BON, Silva RHP, de Abreu AP, Damazio MJ, Melo MM.The Horse Agribusiness Complex is an important activity in the Brazilian agricultural sector. Mangalarga Marchador (MM) is the most numerous breed of horses in Brazil and its temperament and gait characteristics (4-beat stepping gait) allow for the constant growth of the breed and the performance of vs. equestrian activities. The training management currently adopted with MM horses is based on empiricism, and scientific support is necessary to improve the well-being of horses in competitions and maintain the physical integrity of these athletes. Therefore, this study aimed to verify the effect...
Evaluation of the subcapsular technique for primary closure castration in donkeys (Equus asinus).
Scientific reports    July 7, 2021   Volume 11, Issue 1 14080 doi: 10.1038/s41598-021-93585-y
Ibrahim A, Mahmoud UT, Ali MM, Ragab SMM.This study described the subcapsular technique for primary closure castration in donkeys with special regard to its efficiency and welfare impacts. The study was conducted on twelve adult male donkeys, allocated randomly into two groups; subcapsular castration (SC) and open castration (OC) groups, whether the donkeys were subjected to surgical castration either by subcapsular or open castration techniques, respectively. Testosterone, cortisol, lactate, glucose, total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), triglyceride (TG), and nitric oxide (NO) were measured before an...
Equine inflammatory response to abdominal surgery in the absence of gastrointestinal disease.
Journal of veterinary emergency and critical care (San Antonio, Tex. : 2001)    July 1, 2021   Volume 31, Issue 5 601-607 doi: 10.1111/vec.13092
Bowlby C, Mudge M, Schroeder E, Godman J, Hurcombe S.To investigate the, equine inflammatory response to ventral midline celiotomy in the absence of gastrointestinal disease in horses of varying body condition scores primarily using serial measurements of serum amyloid A (SAA). Methods: Experimental clinical study. Methods: University teaching hospital. Methods: Ten adult light breed horses free of any clinical disease, 5 with body condition score (BCS) 3-4/9 and 5 with BCS 7-8/9. Methods: Horses had a ventral midline celiotomy performed under general anesthesia, including manual decompression of the small intestine. SAA, semiquantitative fibrin...
Evaluation of Medetomidine-Ketamine for Immobilization of Feral Horses in Romania.
Frontiers in veterinary science    June 21, 2021   Volume 8 655217 doi: 10.3389/fvets.2021.655217
Roşu O, Melega I, Evans AL, Arnemo JM, Küker S.Feral horses are immobilized for a variety of reasons including population control via contraceptives. Although opioid combinations have been successfully used for immobilization of feral horses, there is a need for combinations using drugs that are more readily available and present less of a human health hazard. We evaluated the chemical immobilization with physiological measurements and blood gas analyses of 91 free-ranging feral horses () remotely immobilized with a combination of 30 mg medetomidine and 775 mg ketamine in a single disposable 6 ml dart. During immobilization, heart rate, re...
Blood lactate concentrations and heart rates of Colombian Paso horses during a field exercise test.
Veterinary and animal science    June 8, 2021   Volume 13 100185 doi: 10.1016/j.vas.2021.100185
Cabrera AMZ, Soto MJC, Aranzales JRM, Valencia NMC, Gutiérrez MPA.Information on performance indices in Paso horses is scarce. Field exercise tests are necessary to recreate the exertion that occurs during training and competition. To describe blood lactate concentrations and heart rates of untrained Colombian Paso horses (CPHs) in response to a field exercise test. A 30-minutes-long standardized field exercise test was carried out on 11 untrained adult CPHs of both sexes. Blood lactate concentration (BLConc) and heart rate (HR) were measured before, during each step of the test, and at recovery. The BLConc and HR were used to calculate the HR at which a BLC...
Influence of endocrine disease on l-lactate concentrations in blood of ponies.
Journal of veterinary internal medicine    May 27, 2021   Volume 35, Issue 3 1582-1588 doi: 10.1111/jvim.16079
Dunkel B, Knowles EJ, Chang YM, Menzies-Gow NJ.Blood l-lactate concentrations are higher in people with developing or established diabetes mellitus and insulin resistance. Objective: To investigate whether blood l-lactate concentrations are positively correlated with measures of insulin dysregulation (ID) or increased autumnal ACTH concentrations in ponies. Methods: Systemically healthy client-owned ponies (n = 101). Methods: Prospective case-control study. Blood samples were obtained from 101 clinically healthy ponies. Breed, weight, height, and subjective and objective measures of body condition were recorded. Blood l-lactate, glucose, t...
Development of a constant pressure perfused ex vivo model of the equine larynx.
PloS one    May 20, 2021   Volume 16, Issue 5 e0251530 doi: 10.1371/journal.pone.0251530
Otto S, Michler JK, Dhein S, Mülling CKW.Distal axonopathy is seen in a broad range of species including equine patients. In horses, this degenerative disorder of the recurrent laryngeal nerve is described as recurrent laryngeal neuropathy (RLN). The dysfunctional innervation of the cricoarytenoideus dorsalis muscle (CAD) leads to a loss of performance in affected horses. In general, ex vivo models of the larynx are rare and for equine patients, just one short report is available. To allow for testing new therapy approaches in an isolated organ model, we examined equine larynges in a constant pressure perfused setup. In order to chec...
Quantifying the Impact of Mounted Load Carrying on Equids: A Review.
Animals : an open access journal from MDPI    May 7, 2021   Volume 11, Issue 5 1333 doi: 10.3390/ani11051333
Bukhari SSUH, McElligott AG, Parkes RSV.There are approximately 112 million working equids in developing countries, many of which are associated with brick kilns. Brick kilns and overloading are associated with welfare problems in working equids. Understanding equids' abilities and influencing factors are important for both effective performance and welfare. Traditionally, measurement of the amount of 'bone' was used, and more recently, gait symmetry has been identified as a potential marker for loading capacity. Assessment of stride parameters and gait kinematics provides insights into adaptations to loading and may help determine ...
Biochemical and hematologic changes in whole blood from Brazilian horses stored in citrate-phosphate-dextrose-adenine pouches for up to 28 days.
Veterinary clinical pathology    April 25, 2021   Volume 50, Issue 2 221-226 doi: 10.1111/vcp.12973
Dorneles TEA, Costa Junior JD, Almeida RM, Teixeira Neto AR.Anaerobic cellular metabolism causes a series of structural and physiologic changes during storage that could compromise post-transfusion viability, reducing the safety of using blood stored for an extended period. Objective: We aimed to follow the biochemical and hematologic alterations of equine blood stored in plastic bags containing citrate-phosphate-dextrose-adenine (CPDA-1) for up to 28 days. Methods: Whole blood samples (450 mL) were collected from 20 Brazilian Saddle horses into CPDA-1 pouches and stored between 2°C and 6°C in a blood bank. On days 0, 7, 14, 21, and 28 of storage, ...
The Physical Activity-Dependent Hematological and Biochemical Changes in School Horses in Comparison to Blood Profiles in Endurance and Race Horses.
Animals : an open access journal from MDPI    April 14, 2021   Volume 11, Issue 4 1128 doi: 10.3390/ani11041128
Maśko M, Domino M, Jasiński T, Witkowska-Piłaszewicz O.Blood testing is one of the most important ways to improve performance, facilitate recovery and monitor the training of endurance and race horses. However, little is known about the physical activity-dependent changes of blood parameters in horses used for pleasure and in riding schools. This study aimed to perform routine blood tests for training monitoring of sport horses in three different horse types of use. Then the values of blood indicators were compared between school, endurance and race horses to find similarities in the physical activity-dependent profile. The study was carried out o...
Impacts of DigestaWell NRG Supplementation on Post Exercise Muscle Soreness in Unconditioned Horses, a Pilot Study.
Journal of equine veterinary science    March 25, 2021   Volume 101 103455 doi: 10.1016/j.jevs.2021.103455
Suagee-Bedore JK, Shen Y, Porr S, Girard ID, Bennett-Wimbush K, Wagner AL.Exercising horses are commonly plagued by muscle fatigue and soreness, which can result in reduced performance ability. In the present study, ten unconditioned horses were fed 200g per day DigestaWell NRG, a commercial dietary supplement containing Yucca schidigera and Trigonella foenum-graecum, two herbs shown in other species to reduce post-exercise muscle pain and soreness. A control, unsupplemented group contained ten horses of similar age, breed, and gender. Horses completed a 50 minutes, ridden standardized exercise test of moderate intensity immediately prior to (Period1) and after 28 d...