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Topic:Equine Medicine

Equine Medicine encompasses the study and application of medical practices specifically related to the health and well-being of horses. This field involves the diagnosis, treatment, and prevention of diseases and disorders affecting equine species. It covers a wide range of topics, including internal medicine, surgery, reproduction, and pharmacology. Research in equine medicine often focuses on understanding the physiological and pathological processes unique to horses, as well as developing effective therapeutic interventions. This page aggregates peer-reviewed research studies and scholarly articles that explore various aspects of equine medicine, providing insights into the latest advancements and methodologies in the care and management of horse health.
Feasibility of transthoracic echocardiographic guidance for multicatheter electrophysiological mapping studies in horses.
Journal of veterinary internal medicine    August 3, 2024   Volume 38, Issue 5 2686-2697 doi: 10.1111/jvim.17156
Vernemmen I, Buschmann E, Demeyere M, Verhaeghe LM, Van Steenkiste G, Decloedt A, van Loon G.Improved characterization of arrhythmias is based on minimally invasive catheterizations. However, these catheterizations have been poorly explored in horses because apart from 3-dimensional (3D) mapping systems, continuous guidance of the catheter's position with adequate detail is difficult using current imaging modalities. Objective: Position multiple electrophysiology catheters simultaneously at predetermined strategical positions in the heart using transthoracic echocardiographic guidance. Methods: Eight adult healthy horses. Methods: Observational study. Two electrophysiological studies ...
‘Chat’ hatches could boost stabled horses’ wellbeing.
The Veterinary record    August 2, 2024   Volume 195, Issue 3 92-93 doi: 10.1002/vetr.4545
Loeb J.Josh Loeb discusses new research suggesting that a small adjustment to stable design could improve welfare for horses.
Return to sport activity following ultrasonographic diagnosis and conservative management of spontaneous injuries of the serratus cervicis ventralis and serratus thoracis ventralis muscles in 11 endurance horses.
Journal of the American Veterinary Medical Association    August 2, 2024   1-6 doi: 10.2460/javma.24.03.0211
Puccetti M, Pilati N, Beccati F, Denoix JM.To describe the clinical diagnosis, ultrasound findings, and outcome of 11 endurance horses with injuries to the serratus ventralis thoracis (SVT) or serratus ventralis cervicis (SVC) muscle. Methods: 11 endurance horses competing in medium- to high-level competitions and presenting with lameness caused by injuries to the SVT or SVC muscle, as confirmed by ultrasonography. Methods: Physical examinations revealed swelling caudal to the shoulder region associated with dorsocranial displacement of the scapula and edema of the ventral thorax for horses with SVT injuries. Swelling cranial to the sc...
The Effects of Intra-Articular Triamcinolone and Autologous Protein Solution on Metabolic Parameters in Horses.
Animals : an open access journal from MDPI    August 2, 2024   Volume 14, Issue 15 2250 doi: 10.3390/ani14152250
Page AE, Johnson M, Parker JL, Jacob O, Poston R, Adams AA, Adam EN.Intra-articular corticosteroids are a popular treatment choice for joint-associated pain and inflammation in horses despite recent work on the metabolic effects of these drugs. The goal of this project was to compare metabolic effects between intra-articular (IA) triamcinolone acetonide (TA) and an autologous protein solution (APS). Five mixed-breed geldings (4-9 years) were utilized for this project. Three identical and consecutive 28-day treatment blocks were used, with metacarpophalangeal IA treatments consisting of equal volumes of saline, a commercially available APS, or 9 mg of TA. Regul...
Emergency Standing Laparoscopic Treatment of Uncontrolled Post-Castration Hemorrhage in Two Geldings.
Animals : an open access journal from MDPI    August 2, 2024   Volume 14, Issue 15 doi: 10.3390/ani14152252
Delvescovo B, McOnie R, Pearson G, Pugliese B, Hackett ES.Persistent hemorrhage of testicular vessels is a potentially life-threatening complication of equine castration. Frequently, general anesthesia is required to retrieve and ligate the bleeding vasculature when standing wound packing and retrieval of the spermatic cord are unsuccessful. We propose standing laparoscopic ligation of the testicular arteries via the paralumbar fossa as a rapid, effective means of halting hemorrhage while avoiding castration site trauma as well as the cardiovascular and recovery risks of general anesthesia. Methods: Two geldings, 6 and 9 months old, presented for eme...
Can the reduced GH, IGF-1, and ovarian steroids concentrations be considered as suspected biomarkers of age-associated functional deficit in mares?
Theriogenology    August 2, 2024   Volume 228 75-80 doi: 10.1016/j.theriogenology.2024.08.001
Satué K, Fazio E, Velasco-Martinez MG, La Fauci D, Barbiera G, Medica P, Cravana C.In humans' and experimental animals' components of the somatotropic axis, such as growth hormone (GH) and insulin-like growth factor 1 (IGF-1) concentrations, decrease with advancing age. Although there is evidence regarding IGF-1, the effect of age on GH in mares, as well as the relationships between both parameters, have not yet been elucidated. On the other hand, although GH and IGF-1 are related to follicular development, it is unknown if they could be correlated with the circulating concentrations of ovarian steroids in mares, as occurs in other species. The hypothesis of this study was t...
Molecular testing for equine herpesviruses 1 (EHV-1) in healthy postpartum broodmares.
The Canadian veterinary journal = La revue veterinaire canadienne    August 2, 2024   Volume 65, Issue 8 813-816 
Arroyo LG, Gomez DE, Moore A, Papapetrou M, Lillie BN.Our objective was to determine whether equine herpesviruses 1 (EHV-1) viral nucleic acids could be detected immediately after foaling from nasal and vaginal swabs, whole blood, and placental tissue of healthy mares. Unassigned: Nasal and vaginal swabs, EDTA blood, and placental tissue (296 samples) were collected from 74 clinically healthy postpartum broodmares within 24 h after giving birth to live, clinically healthy foals. All samples were tested (PCR) for nucleic acids of neuropathogenic and non-neuropathogenic strains of EHV-1, and all were negative. Unassigned: As EHV-1 was not detected ...
Investigation of Extracted Plasma Cell-Free DNA as a Biomarker in Foals with Sepsis.
Veterinary sciences    August 1, 2024   Volume 11, Issue 8 346 doi: 10.3390/vetsci11080346
Hobbs KJ, Cooper BL, Dembek K, Sheats MK.Cell-free DNA (cfDNA) is fragmented extracellular DNA that is released from cells into various body fluids. Previously published data from adult horses supports cfDNA as a potential disease biomarker, but also shows that direct measurement in plasma is inaccurate due to matrix effect. It is currently unknown whether a similar matrix effect exists in foal plasma. Given this, the objectives of the current study were to investigate foal plasma for potential matrix effect during fluorescence measurement of cfDNA using a Qubit fluorometer, and to determine whether neat and/or extracted plasma cfDNA...
Histological and chemical view on parotid duct sialolithiasis in the Slovakian warmblood mare.
Veterinary research communications    July 31, 2024   doi: 10.1007/s11259-024-10485-y
Korim F, Revajová V, Koľvek F, Bujňák L, Hreus S, Všianský D.The parotid duct has been reported to be the most common site of sialoliths in horses. In this case report, we described the first confirmed case of the equine sialolithiasis in Slovakia. The work was aimed to describe the transcutaneous approach to removing the sialolith, which manifested as a hard painless mass in the area of the maxillary cheek teeth, in a 14-year-old Slovakian warmblood mare. Pathological-anatomical and histological examination after extirpation confirmed the presence of parotid duct ectasia resulting from calculus. The mineral composition of the sialolith was determined w...
Fibrobacter sp. HC4, a newly isolated strain, demonstrates a high cellulolytic activity as revealed by enzymatic measurements and in vitro assay.
Applied and environmental microbiology    July 31, 2024   Volume 90, Issue 8 e0051424 doi: 10.1128/aem.00514-24
Froidurot A, Jacotot E, Julliand S, Grimm P, Julliand V.Despite their low quantity and abundance, the cellulolytic bacteria that inhabit the equine large intestine are vital to their host, as they enable the crucial use of forage-based diets. Fibrobacter succinogenes is one of the most important intestinal cellulolytic bacteria. In this study, Fibrobacter sp. HC4, one cellulolytic strain newly isolated from the horse cecum, was characterized for its ability to utilize plant cell wall fibers. Fibrobacter sp. HC4 consumed only cellulose, cellobiose, and glucose and produced succinate and acetate in equal amounts. Among genes coding for CAZymes, 26% o...
Intracardiac ultrasound-guided transseptal puncture in horses: Outcome, follow-up, and perioperative anticoagulant treatment.
Journal of veterinary internal medicine    July 31, 2024   Volume 38, Issue 5 2707-2717 doi: 10.1111/jvim.17158
Vernemmen I, Buschmann E, Van Steenkiste G, Demeyere M, Verhaeghe LM, De Somer F, Devreese KMJ, Schauvliege S, Decloedt A, van Loon G.Cardiac catheterizations in horses are mainly performed in the right heart, as access to the left heart traditionally requires an arterial approach. Transseptal puncture (TSP) has been adapted for horses but data on follow-up and closure of the iatrogenic atrial septal defect (iASD) are lacking. Objective: To perform TSP and assess postoperative complications and iASD closure over a minimum of 4 weeks. Methods: Eleven healthy adult horses. Methods: Transseptal puncture was performed under general anesthesia. Serum cardiac troponin I concentrations were measured before and after puncture. Wee...
Quantitative microCT imaging of a whole equine placenta and its blood vessel network.
Placenta    July 31, 2024   Volume 154 216-219 doi: 10.1016/j.placenta.2024.07.313
Laundon D, Proudley E, Basford PJ, Katsamenis OL, Chatelet DS, Cleal JK, Gostling NJ, Chavatte-Palmer P, Lewis RM.Placental structure is linked to function across morphological scales. In the placenta, changes to gross anatomy, such as surface area, volume, or blood vessel arrangement, are associated with suboptimal physiological outcomes. However, quantifying each of these metrics requires different laborious semi-quantitative methods. Here, we demonstrate how, with minimal sample preparation, whole-organ computed microtomography (microCT) can be used to calculate gross morphometry of the equine placenta and a range of additional metrics, including branching morphometry of placental vasculature, non-dest...
Transcorneal aspiration for management of primary iris cysts in the standing horse.
Veterinary medicine and science    July 30, 2024   Volume 10, Issue 5 e1570 doi: 10.1002/vms3.1570
Conduit S, Bowen M, Hallowell G, Pereira R, Rapezzano G, Redpath A.Equine primary iris cysts are usually incidental findings but, if associated with clinical signs, may require intervention. The use of laser (Nd:Yag or diode) has been reported but requires specialised equipment. Transcorneal aspiration has not been previously evaluated in the standing horse. To review outcomes of standing transcorneal aspiration of primary iris cysts (STAPIC) in horses. Horses were identified from electronic patient records from 2018 to 2024 across four collaborating centres. Clinical presentation and outcomes were identified and reported using descriptive statistics. Eightee...
Prevalence of movement asymmetries in high-performing riding horses perceived as free from lameness and riders’ perception of horse sidedness.
PloS one    July 30, 2024   Volume 19, Issue 7 e0308061 doi: 10.1371/journal.pone.0308061
Zetterberg E, Persson-Sjodin E, Lundblad J, Hernlund E, Rhodin M.A high proportion of horses in training, perceived as free from lameness by their owner, exhibit vertical movement asymmetries. These types of asymmetries are sensitive measures of lameness, but their specificity as indicators of orthopaedic pathology or locomotor function remains unclear. Equine athletes performing at a high level could be assumed to exhibit a higher degree of movement symmetry compared with the general horse population, but this has not been confirmed. This study investigated the prevalence of movement asymmetries in horses performing at a high level in three equestrian disc...
Application of blood parameters for the early diagnosis of natural ascending placentitis in pregnant mares.
Theriogenology    July 30, 2024   Volume 228 37-53 doi: 10.1016/j.theriogenology.2024.07.025
Feijo LS, Wolfsdorf KE, Canisso IF, Parry S, Felippe MJB.Placental infection is an important cause of late-term pregnancy loss and neonatal diseases in horses. Detection of changes in blood parameters especially during early placentitis could improve the diagnostic accuracy, treatment decision, and potential outcomes. The objectives of this 2-part study were to identify differences in circulating immunological, inflammatory, and hormonal parameters between mares with natural ascending placentitis and control mares; evaluate each and combination of parameters as predictors of placentitis; and determine how these parameters indicate severity of placen...
Development and validation of general plasma screening method for performance enhancing drugs in racehorses utilizing liquid chromatography-high-resolution mass spectrometry (LC-HRMS).
Drug testing and analysis    July 29, 2024   doi: 10.1002/dta.3774
Lomnicka I, Dubey S, Waller P, Vora D, Dirikolu L.The screening of drugs in plasma and urine often requires initial extraction (such as liquid-liquid extraction and solid-phase extraction) before the samples are submitted to instrumental analyses. These extraction procedures are often laborious and time-consuming. In this manuscript, a high-throughput automated assay based on liquid chromatography-high-resolution mass spectrometry (LC-HRMS) suitable for use as an initial testing procedure covering multiple classes of compounds prohibited in horse racing is described. The assay requires a 600-μL plasma aliquot, which is subjected to solid pha...
Carbohydrate digestion in the stomach of horses grazed on pasture, fed hay or hay and oats.
Journal of equine veterinary science    July 27, 2024   Volume 141 105152 doi: 10.1016/j.jevs.2024.105152
Bachmann M, Schusser GF, Wensch-Dorendorf M, Pisch C, Bochnia M, Santo MM, Netzker H, Woitow G, Thielebein J, Kesting S, Riehl G, Greef JM....Concentrations of starch, mono- and disaccharides, fructans, hemicellulose and cellulose were analysed in feed and gastric digesta of horses in relation to acid insoluble ash as a marker indigestible in the stomach. Twenty-four horses were allocated to pasture 24 h/d (PST; n = 4), hay ad libitum (HAY; n = 8), hay ad lib. and oats at 1 g starch/kg body weight (BWT)/meal (OS1; n = 6) and hay ad lib. and oats at 2 g starch/kg BWT/meal (OS2; n = 5). One horse was excluded from the analysis. The horses were fed the ration a minimum of 34 days. Following euthanasia and dissection, digesta was sample...
Use of excised ovaries for oocyte recovery by ultrasound guided follicular aspiration – Validation of an experimental model for research purposes in live mares ovum pick up.
Theriogenology    July 26, 2024   Volume 228 9-16 doi: 10.1016/j.theriogenology.2024.07.023
Sala-Ayala L, Pytel AT, Stychno K, Cuervo-Arango J.Ovum pick-up (OPU) by transvaginal ultrasound guided follicle aspiration in mares is a common assisted reproductive technique used for oocyte recovery and in vitro production of horse embryos. There has been relatively little research into the factors influencing oocyte recovery in OPU from live mares. The objective of this study was to compare oocyte recovery and morphology of ultrasound-guided follicle puncture and aspiration in live mares and in postmortem excised ovaries, in order to validate an experimental model for research purposes of the efficiency of OPU in mares. Data from OPU perfo...
Technique for guttural pouch bead removal using a novel three-dimensional (3D)-printed instrument.
Veterinary surgery : VS    July 26, 2024   Volume 53, Issue 7 1219-1230 doi: 10.1111/vsu.14141
Cardona GC, Dahlgren LA, Byron CR, McKenzie HC, Werre SR, Bogers SH.The aim of the present study was to determine if a three-dimensional (3D)-printed instrument technique would improve lavage removal of plastic beads (guttural pouch [GP] chondroid mimics) through a dorsal pharyngeal recess (DPR) fenestration. We hypothesized that using a 3D-printed instrument placed through the DPR fenestration would remove more beads, reduce lavage time and incur less soft tissue damage than using a lavage tube control or instrument placement through the salpingopharyngeal ostium (SPO). Methods: Experimental cadaveric study. Methods: A total of 30 cadaveric equine heads. Meth...
Single injection of intra-articular autologous protein solution in horses with acute interleukin-1B-induced synovitis decreases joint pathology scores.
Equine veterinary journal    July 25, 2024   doi: 10.1111/evj.14203
Usimaki A, Ciamillo SA, Barot D, Linardi RL, Engiles JB, Ortved KF.Autologous protein solution (APS) has been shown to decrease lameness in horses with osteoarthritis (OA). Synovitis is an early driver of OA, providing an opportunity to intervene in the progression of disease via intra-articular (IA) therapeutics. Objective: The objective of this study was to investigate the effects of a single IA APS injection in horses with interleukin-1β (IL-1β)-induced synovitis. We hypothesised that APS would decrease joint swelling and lameness, improve synovial fluid parameters and improve joint pathology scores in horses compared with untreated controls. Methods: Ra...
Influence of climatic changes on respiratory health in a teaching herd of outdoor-housed horses.
Veterinary journal (London, England : 1997)    July 25, 2024   Volume 307 106198 doi: 10.1016/j.tvjl.2024.106198
Calomeno SS, Freitas Santi T, Barbosa B, Weber SH, Oliveira TM, Machado GF, Michelotto PV.Horses maintained outdoors may experience a lower-allergenic environment compared to their stabled counterparts. This study hypothesizes that climatic changes in southern Brazil can influence respiratory status. To test this hypothesis, we evaluated the lower airways of 17 horses from a teaching herd in southern Brazil, maintained outdoors, during winter, spring, and summer. Except for one mare with a history of severe asthma, all horses were considered healthy and underwent a physical examination. Airway endoscopic evaluation included scoring for tracheal mucus (0-5) and bronchial septum thic...
Inertial Sensor-Based Quantification of Movement Symmetry in Trotting Warmblood Show-Jumping Horses after “Limb-by-Limb” Re-Shoeing of Forelimbs with Rolled Rocker Shoes.
Sensors (Basel, Switzerland)    July 25, 2024   Volume 24, Issue 15 doi: 10.3390/s24154848
Bark C, Reilly P, Weller R, Pfau T.Hoof care providers are pivotal for implementing biomechanical optimizations of the musculoskeletal system in the horse. Regular visits allow for the collection of longitudinal, quantitative information ("normal ranges"). Changes in movement symmetry, e.g., after shoeing, are indicative of alterations in weight-bearing and push-off force production. Ten Warmblood show jumping horses (7-13 years; 7 geldings, 3 mares) underwent forelimb re-shoeing with rolled rocker shoes, one limb at a time ("limb-by-limb"). Movement symmetry was measured with inertial sensors attached to the head, withers, and...
Roles of collagen cross-links and osteon collagen/lamellar morphotypes in equine third metacarpals in tension and compression tests.
The Journal of experimental biology    July 24, 2024   Volume 227, Issue 14 jeb247758 doi: 10.1242/jeb.247758
Skedros JG, Dayton MR, Cronin JT, Mears CS, Bloebaum RD, Wang X, Bachus KN.Many bones experience bending, placing one side in net compression and the other in net tension. Because bone mechanical properties are relatively reduced in tension compared with compression, adaptations are needed to reduce fracture risk. Several toughening mechanisms exist in bone, yet little is known of the influences of secondary osteon collagen/lamellar 'morphotypes' and potential interplay with intermolecular collagen cross-links (CCLs) in prevalent/predominant tension- and compression-loaded regions. Paired third metacarpals (MC3s) from 10 adult horses were prepared for mechanical test...
Fungal aerosol and particulate matter in horse stables in Poland.
Applied microbiology and biotechnology    July 24, 2024   Volume 108, Issue 1 426 doi: 10.1007/s00253-024-13258-4
Grzyb J, Podstawski Z, Bulski K.Horses stay in different types of stables; especially during the cold season, they stay inside for most of the day. A stable is also a place where many people spend quite a lot of time either as employees who care for and train horses or as equine enthusiasts. Keeping horses in stables causes their constant exposure to high concentrations of particulate matter (PM) and molds in the air inside these facilities. The study was conducted in Udórz Stud Farm located in the southern region of Poland. It was carried out in two different types of stables: three runners and two box stables. The study c...
Causes, treatment, and outcome of hyphema, fibrin, and vitreal hemorrhage in horses: 219 cases (2012-2023).
Journal of the American Veterinary Medical Association    July 24, 2024   Volume 262, Issue S2 S94-S101 doi: 10.2460/javma.24.04.0258
Gould J, Wells M, de Linde Henriksen M, Gilger BC.To determine the causes, time to resolution, effect of therapeutics, and ocular sequelae of hyphema, fibrin, and/or vitreal hemorrhage in horses. Methods: 225 horses (219 eyes) who were diagnosed with hyphema, fibrin, and/or vitreal hemorrhage. Methods: Records were retrospectively reviewed for the horses. Signalment, ophthalmic examination findings, causes, treatments, and outcomes were evaluated. Results: Common causes of intraocular fibrin or hemorrhage were equine recurrent uveitis (42/219 horses), corneal stromal abscess (32/219 horses), corneal stromal ulcer (25/219 horses), and trauma (...
An Explorative Anatomical Study on Inter-Individual Variation of the Tibial Nerve and Landmarks for Perineural Anesthesia in Horses.
Animals : an open access journal from MDPI    July 24, 2024   Volume 14, Issue 15 2161 doi: 10.3390/ani14152161
De Schryver M, Oosterlinck M.Perineural anesthesia of the tibial nerve can be performed ultrasound-guided or blindly, with the latter still being commonly used in equine practice due to practical constraints, despite its lower accuracy and hence, common failure to achieve desensitization. This may be associated with anatomical variations or inadequate landmarks for injection. To examine the course of the tibial nerve, document potential anatomical variations, and determine optimal landmarks for perineural injection, dissection was conducted along the medial aspect of the tibia in 10 paired cadaver hindlimbs. No anatomical...
Time budgets and weight shifting as indicators of pain in hospitalized horses.
Frontiers in pain research (Lausanne, Switzerland)    July 23, 2024   Volume 5 1410302 doi: 10.3389/fpain.2024.1410302
Nowak M, Martin-Cirera A, Jenner F, Auer U.Pain assessment in horses presents a significant challenge due to their nonverbal nature and their tendency to conceal signs of discomfort in the presence of potential threats, including humans. Therefore, this study aimed to identify pain-associated behaviors amenable to automated AI-based detection in video recordings. Additionally, it sought to determine correlations between pain intensity and behavioral and postural parameters by analyzing factors such as time budgets, weight shifting, and unstable resting. The ultimate goal is to facilitate the development of AI-based quantitative tools f...
Concurrent radiological and ultrasonographical findings in the forelimb proximal sesamoid bones and adjacent suspensory ligament branches in yearling and 2-year-old Thoroughbred sales horses.
Equine veterinary journal    July 22, 2024   doi: 10.1111/evj.14120
Peat FJ, Kawcak CE, McIlwraith CW, Berk JT, Keenan DP.Changes in the proximal sesamoid bones (sesamoids) and the insertional region of the adjacent suspensory ligament branch (branch) are of particular importance in young Thoroughbreds sold at public auction. Little is known about the prevalence of concurrent ultrasonographical branch change, relative to the various grades of radiological sesamoid appearance. Objective: To examine the existence of concurrent radiological and ultrasonographical findings in individual sesamoid-branch units in sales horses; to determine whether there are any radiological findings that are consistently accompanied by...
The use of serum paraoxonase-1 to assess inflammation in horses with colitis.
Veterinary clinical pathology    July 21, 2024   Volume 53, Issue 3 358-368 doi: 10.1111/vcp.13372
Winther MF, Johnsson J, Madsen PK, Pihl TH, Paltrinieri S, Cerón JJ, Scavone D, Pardo-Marin L, Jacobsen S.Paraoxonase-1 (PON-1) has been suggested as a marker of inflammation and oxidative stress in horses and could potentially be used for prognostication in horses with colitis. Objective: Assessment of PON-1 in horses with colitis and comparison of two methods. Methods: Serum PON-1 was measured by two methods (paraoxon and p-nitrophenyl acetate) in 161 horses with colitis and 57 controls. Follow-up samples obtained during hospitalization were available from 106 horses with colitis. The two methods were compared. Results: Serum PON-1 was significantly lower in horses with colitis than in healthy h...
Impact of low-dose ozone supplementation on motility parameters and bacterial growth in horse cryopreserved semen.
Veterinary and animal science    July 21, 2024   Volume 25 100380 doi: 10.1016/j.vas.2024.100380
Fanelli D, Moroni R, Sala G, Melanie P, Tarabella I, Telleschi N, Maltinti S, Giorgi M, Barsotti G, Passamonti F, Marmorini P, Rota A, Camillo F....Two studies were conducted to evaluate the use of medical ozone (O) in commercial extenders for equine semen cryopreservation. In the first study (Study 1), 0, 5, and 15 µg/mL of O were added to diluents of refrigerated or frozen semen. Samples were evaluated for sperm kinematics at different time points for the chilled samples and after a thermoresistence test for the frozen/thawed samples. In the second study (Study 2), 0, 5, and 10 µg/mL of O were added to an antibiotic-free diluent for refrigerated semen for comparison with the control group in which semen was diluted in the same diluent...
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