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Topic:Clinical Study

Clinical studies in equine research involve the systematic investigation of health and disease in horses through structured scientific methods. These studies aim to evaluate the safety and efficacy of treatments, understand disease mechanisms, and improve veterinary care practices. Clinical studies can include randomized controlled trials, observational studies, and case-control studies. They may focus on various aspects such as pharmacokinetics, therapeutic interventions, and diagnostic techniques. This page compiles peer-reviewed research studies and scholarly articles that explore the design, implementation, and findings of clinical studies in equine medicine, providing insights into their impact on horse health and veterinary practices.
Effects of insulin on IGF-1 receptors in equine lamellar tissue in vitro.
Domestic animal endocrinology    July 29, 2020   Volume 74 106530 doi: 10.1016/j.domaniend.2020.106530
Rahnama S, Spence R, Vathsangam N, Baskerville CL, Bailey SR, de Laat MA, Anderson ST, Pollitt CC, Sillence MN.Although it is understood that equine endocrinopathic laminitis can be triggered by high concentrations of insulin, it is unclear whether this represents a direct action on lamellar tissue via insulin receptors (InsR), an interaction with IGF-1 receptors (IGF-1R), or some other, indirect action. This uncertainty is because of the reported scarcity of InsR in lamellar tissue and the low affinity of insulin for equine IGF-1R. In the present study, the effects of insulin and IGF-1 (as a positive control) were examined using lamellar explants isolated from the hooves of healthy horses and incubate...
Effects of pituitary pars intermedia dysfunction and Prascend (pergolide tablets) treatment on endocrine and immune function in horses.
Domestic animal endocrinology    July 29, 2020   Volume 74 106531 doi: 10.1016/j.domaniend.2020.106531
Miller AB, Loynachan AT, Bush HM, Hart KA, Barker VD, Campana-Emard AG, Grubbs ST, Adams AA.It remains unclear how pituitary pars intermedia dysfunction (PPID) and pergolide treatment (Prascend [pergolide tablets]) affect endocrine and immune function in horses. To evaluate these effects, blood was collected regularly from 28 university-owned horses (10 Non-PPID, 9 PPID control [PC], and 9 PPID treatment [PT]) over approximately 15 mo. Pergolide treatment was initiated after Day 0 collections. Analyses included ACTH, insulin, total cortisol, free cortisol, complete blood counts, plasma myeloperoxidase, and cytokine/receptor gene expression in basal whole blood and in vitro stimulat...
Variability, repeatability and test-retest reliability of equine flash visual evoked potentials (FVEPs).
BMC veterinary research    July 29, 2020   Volume 16, Issue 1 261 doi: 10.1186/s12917-020-02463-8
Ström L, Bröjer J, Ekesten B.Visual evoked potentials (VEPs) are electrical potentials generated by neurons in the central nervous system in response to visual stimuli. A series of positive and negative wavelets in response to flash-stimuli (flash-VEP; FVEP) or reversing, iso-luminant patterns (pattern-VEP; PVEP) are recorded. Pathological conditions affecting the post-retinal pathways can alter overall waveform morphology, and also affect wavelet peak times and amplitudes. FVEPs have recently been described in horses, but more data on the variability within and between subjects is required, to adequately interpret result...
Infrared methodologies for the assessment of skin temperature daily rhythm in two domestic mammalian species.
Journal of thermal biology    July 29, 2020   Volume 92 102677 doi: 10.1016/j.jtherbio.2020.102677
Giannetto C, Arfuso F, Giudice E, Gianesella M, Fazio F, Panzera M, Piccione G.To assess the accuracy of infrared methodologies for daily rhythm monitoring of skin temperature, five clinically healthy Italian Saddle gelding horses, and five not pregnant and not lactating Camosciata goats, were monitored every 4 h over a 48 h period. The horses were housed in individual boxes, while the goats in two indoor pens, under natural photoperiod and natural environmental temperature. In each animal, skin temperature was recorded with the use of a digital infrared camera and a non-contact infrared thermometer, in five regions: neck, shoulder, ribs, flank and croup. Recorded valu...
Computed Tomography (CT)-Assisted 3D Cephalometry in Horses: Interincisal Angulation of Clinical Crowns.
Frontiers in veterinary science    July 29, 2020   Volume 7 434 doi: 10.3389/fvets.2020.00434
Kau S, Failing K, Staszyk C.The angle encompassed between opposing incisors in horses is assumed to decline with age. Previous studies merely consider the overall profile view of clinical crowns presuming a generalized angle, neglecting potential tooth position-dependent differences. Cephalometric measurements from 3D computed tomographic thick-slab reconstructions of single incisors within a global reference frame were used to determine clinical crown interincisal angulation (IIA) of 48 horses. Based on predefined dentoalveolar landmarks, IIA was defined as the angle enclosed by the respective labial axis of the clinica...
[Effects of pony-assisted therapy on the behavior of older people with neurocognitive disorders living in nursing homes].
Geriatrie et psychologie neuropsychiatrie du vieillissement    July 28, 2020   Volume 18, Issue 3 273-282 doi: 10.1684/pnv.2020.0854
Carillon B, Jarzebowski W, Fulpin B, Miliotis A, Bernadac A, Saint-Georges Chaumet Y, Belmin J.Experienced in psychomotricity in two nursing homes, pony-assisted therapy shows first positive effects on psycho-behavioral symptoms. Recognized in the field of disability and psychiatry, this form of therapy is not developed in gerontology. To measure the effects of this therapy on the behavior of elderly people with neurocognitive disorders living in long-term care and especially on the involvement in this activity. Anxiety and apathy were chosen as the measurement criteria. An intervention study with an experimental cross-study design was conducted over two months for 23 subjects. An inter...
Retrospective case series to identify the most common conditions seen ‘out-of-hours’ by first-opinion equine veterinary practitioners.
The Veterinary record    July 27, 2020   Volume 187, Issue 10 404 doi: 10.1136/vr.105880
Bowden A, Boynova P, Brennan ML, England GCW, Mair TS, Furness WA, Freeman SL, Burford JH.The study aim was to describe conditions seen 'out-of-hours' in equine practice. This was a retrospective case series of first opinion 'out-of-hours' cases seen at two equine practices between 2011 and 2013. Data were retrieved on case presentation, diagnostic testing, treatment administered and outcome, and diseases categorised using a systems-based coding system. A hierarchical logistic regression, formulated using a generalised linear model, was used to identify clinical variables associated with a binary outcome of 'critical' cases (required hospitalisation or euthanasia or died). Data fro...
Fractionated semen collection as a tool to rescue fertility in stallions with seminal vesiculitis.
Theriogenology    July 27, 2020   Volume 157 110-120 doi: 10.1016/j.theriogenology.2020.07.014
Oliveira SN, Andrade LRP, Silva LFMC, Araujo EAB, Rayashi RM, Segabinazzi LGTM, Alvarenga MA, Dell'Aqua CP, Dell'Aqua JA, Papa FO.Treatments for seminal vesiculitis have poor outcomes in stallions; thus, the development of alternative strategies is warranted. This study aimed to evaluate fractionated semen collection as a method to restore the fertility of stallions diagnosed with seminal vesiculitis. Eighteen ejaculates from six stallions (three ejaculates/stallion) diagnosed with seminal vesiculitis were harvested in fractions, as follows: Fraction A (FA), the first two jets; Fraction B (FB), the third and fourth jets; and Fraction C (FC), the fifth and remaining jets of the ejaculate. All fractions were subject to sta...
Catheter-based electrical interventions to study, diagnose and treat arrhythmias in horses: From refractory period to electro-anatomical mapping.
Veterinary journal (London, England : 1997)    July 24, 2020   Volume 263 105519 doi: 10.1016/j.tvjl.2020.105519
van Loon G, Van Steenkiste G, Vera L, Decloedt A.Minimally-invasive catheter-based interventional cardiology is a mainstay for the diagnosis and treatment of arrhythmias in human medicine. Very accurate imaging using fluoroscopy, CT and MRI is essential during interventional cardiology procedures. Because these imaging techniques are either not possible or provide too little anatomical detail in horses, echocardiography is currently the best technique to visualize catheters in horses. Over the past decades, catheter-based techniques have been applied to induce arrhythmias using pacing and to perform arrhythmia research using electrophysiolog...
Ex vivo modeling of the airflow dynamics and two-and three-dimensional biomechanical effects of suture placements for prosthetic laryngoplasty in horses.
American journal of veterinary research    July 24, 2020   Volume 81, Issue 8 665-672 doi: 10.2460/ajvr.81.8.665
Lynch NP, Jones SA, Bazley-White LG, Wilson ZF, Raffetto J, Pfau T, Cheetham J, Perkins JD.To identify the degree of left arytenoid cartilage (LAC) abduction that allows laryngeal airflow similar to that in galloping horses, assess 2-D and 3-D biomechanical effects of prosthetic laryngoplasty on LAC movement and airflow, and determine the influence of suture position through the muscular process of the arytenoid cartilage (MPA) on these variables. Methods: 7 equine cadaver larynges. Methods: With the right arytenoid cartilage maximally abducted and inspiratory airflow simulated by vacuum, laryngeal airflow and translaryngeal pressure and impedance were measured at 12 incremental LAC...
Topical Ganciclovir Reduces Viral Excretion in Mares With Equine Coital Exanthema.
Journal of equine veterinary science    July 24, 2020   Volume 94 103199 doi: 10.1016/j.jevs.2020.103199
Vissani MA, Perglione CO, Zabal O, Alvarez G, Thiry E, Barrandeguy M, Parreño V.Equid alphaherpesvirus 3 (EHV-3) is the etiological agent of equine coital exanthema (ECE). Because no vaccines or antiviral therapies are available, prevention consists of clinical examination of mares and stallions before mating or semen collection and resting from breeding activities when lesions are present. However, this methodology does not identify subclinically infected animals. Ganciclovir is the most potent compound known to reduce EHV-3 replication. This study aimed to evaluate the efficacy of topical ganciclovir application to reduce EHV-3 replication in experimentally infected mar...
A retrospective evaluation of the effect of perianesthetic hydromorphone administration on the incidence of postanesthetic signs of colic in horses.
Veterinary anaesthesia and analgesia    July 23, 2020   Volume 47, Issue 6 757-762 doi: 10.1016/j.vaa.2020.06.003
Skrzypczak H, Reed R, Barletta M, Quandt J, Sakai D.To describe the incidence of postanesthetic signs of colic (PASC) in horses and determine if perianesthetic administration of hydromorphone was associated with an increased risk of PASC. Methods: Retrospective, cohort study. Methods: A total of 409 horses. Methods: Anesthesia and clinical records of horses admitted for various procedures from July 2018 to September 2019 were reviewed. Signs of colic and interventions were recorded up to 48 hours after anesthesia. A binomial logistic regression model was used to evaluate the association between the type of surgery, administration of hydromorpho...
Effect of the p38 MAPK inhibitor doramapimod on the systemic inflammatory response to intravenous lipopolysaccharide in horses.
Journal of veterinary internal medicine    July 23, 2020   Volume 34, Issue 5 2109-2116 doi: 10.1111/jvim.15847
Bauquier J, Tudor E, Bailey S.Doramapimod, a p38 MAPK inhibitor, is a potent anti-inflammatory drug that decreases inflammatory cytokine production in equine whole blood in vitro. It may have benefits for treating systemic inflammation in horses. Objective: To determine whether doramapimod is well tolerated when administered IV to horses, and whether it has anti-inflammatory effects in horses in a low-dose endotoxemia model. Methods: Six Standardbred horses. Methods: Tolerability study, followed by a blinded, randomized, placebo-controlled cross-over study. Horses were given doramapimod, and clinical and clinicopathologica...
The use of the geometric morphometric method to illustrate shape difference in the skulls of different-aged horses.
Veterinary research communications    July 23, 2020   Volume 44, Issue 3-4 137-145 doi: 10.1007/s11259-020-09779-8
Liuti T, Dixon PM.The geometric morphometrics method (GMM) is a technique to study scale and shape relationships of structures using Cartesian geometric coordinates rather than linear, areal (of area), or volumetric variables. GMM has been of great value in many biological studies, but does not appear to have been used to examine equine skulls.In this exploratory study, twenty-nine normal equine heads of three different age groups:  16 years old (N = 10) were examined.Computed tomography (CT) bone window DICOM images were reconstructed into isosurfaces (3-dimensional contoured surfaces), onto which landm...
Novel Equine Faecal Egg Diagnostics: Validation of the FECPAKG2.
Animals : an open access journal from MDPI    July 23, 2020   Volume 10, Issue 8 1254 doi: 10.3390/ani10081254
Tyson F, Dalesman S, Brophy PM, Morphew RM.Faecal egg counts (FECs) are the standard method of diagnosing the level of parasitic helminth egg shedding in horses and other grazing animals. Testing before treatment is an important factor in slowing the appearance of anthelmintic resistance in nematode parasites. The FECPAK, optimised for livestock, is reported to allow owners to perform FECs on their own animals without the need for a separate microscope or any specialist knowledge by tapping into remote expertise. However, the performance of the FECPAK has yet to be assessed for equids. Therefore, a comparison of the FECPAK (G2) method ...
Optimizing corneal riboflavin administration in ex vivo horse, dog, rabbit, and pig samples for use in corneal collagen cross-linking.
Veterinary ophthalmology    July 23, 2020   Volume 23, Issue 5 840-848 doi: 10.1111/vop.12807
Zibura AE, Cullen MA, Rutledge H, Lassalle L, Salmon JH, Gilger BC, Westermeyer HD.Determine optimal iontophoresis times for riboflavin delivery to the corneal stroma across different species and compare these to corneal injection. Methods: Ex vivo horse, dog, rabbit, and pig globes were treated with riboflavin administered with either iontophoresis for 2.5-20 minutes with or without corneal epithelium; or with purpose-designed precise corneal injection (PCI) application with intact epithelium. Immediately following riboflavin administration, samples were harvested, frozen, and sectioned. Riboflavin penetration was imaged using fluorescence microscopy. Results: Horse sample...
Atrial fibrillation in horses part 1: Pathophysiology.
Veterinary journal (London, England : 1997)    July 23, 2020   Volume 263 105521 doi: 10.1016/j.tvjl.2020.105521
Decloedt A, Van Steenkiste G, Vera L, Buhl R, van Loon G.Atrial fibrillation (AF) is the most common clinically relevant arrhythmia in horses, with a reported prevalence up to 2.5%. The pathophysiology has mainly been investigated in experimental animal models and human medicine, with limited studies in horses. Atrial fibrillation results from the interplay between electrical triggers and a susceptible substrate. Triggers consist of atrial premature depolarizations due to altered automaticity or triggered activity, or local (micro)reentry. The arrhythmia is promoted by atrial myocardial ion channel alterations, Ca2+ handling alterations, structural ...
Effect of Caudal Traction on Mechanical Nociceptive Thresholds of Epaxial and Pelvic Musculature on a Group of Horses With Signs of Back Pain.
Journal of equine veterinary science    July 23, 2020   Volume 93 103197 doi: 10.1016/j.jevs.2020.103197
Long K, McGowan CM, Hyytiäinen HK.Direct muscular attachment from lumbar vertebrae to the caudal vertebrae of the tail suggests that caudal traction, also described as a tail pull, may affect lumbar vertebral segments and/or associated soft tissues in horses. Traction is a commonly used human manual therapy technique used for pain relief and anecdotally observed to relieve pain in horses. However, research is lacking validating the efficacy of manual caudal traction on the horse. The objective of this study was to determine if caudal traction has an effect on mechanical nociceptive thresholds (MNTs) in a group of horses with c...
Prevalence of Radiographic Signs of Osteoarthritis in Lusitano Purebred Horses.
Journal of equine veterinary science    July 23, 2020   Volume 94 103196 doi: 10.1016/j.jevs.2020.103196
Ramos S, Pinto A, Cardoso M, Alexandre N, Bettencourt E, Monteiro S, Gama LT.Radiographic prevalence and correlation of radiographic findings has not been performed in Lusitano Purebred horses. The aim of this study was to (1) evaluate the prevalence of primary osteoarthritis radiographic findings in Lusitano Purebred horses; (2) to assess correlations between radiographic findings in different joints of the same limb and different limbs; and (3) elucidate the effect of age in the radiographic findings. A radiographic protocol of the stifle, tarsi, fetlocks and distal limbs was done in 98 Lusitanos and the classification of the radiographs was performed using a 0-4 sc...
Degenerative Lumbar Spinal Stenosis.
Revista brasileira de ortopedia    July 23, 2020   Volume 56, Issue 1 9-17 doi: 10.1055/s-0040-1712490
Hennemann S, de Abreu MR.Degenerative lumbar spinal stenosis is the most frequent cause of low back pain and/or sciatica in the elderly patient. Epidemiology, pathophysiology, clinical manifestations and testing are reviewed in a wide current bibliographic investigation. The importance of the relationship between clinical presentation and imaging study, especially magnetic resonance imaging (MRI), is emphasized. Prior to treatment indication, it is necessary to identify the precise location of pain, as well as the differential diagnosis between neurological and vascular lameness. Conservative treatment combining medic...
Retrospective analysis of 15 years of horse-related maxillofacial fracture data at a major German trauma center.
European journal of trauma and emergency surgery : official publication of the European Trauma Society    July 22, 2020   Volume 48, Issue 4 2539-2546 doi: 10.1007/s00068-020-01450-w
Stier R, Tavassol F, Dupke C, Rüter M, Jehn P, Gellrich NC, Spalthoff S.The purpose of this study was to estimate the effect of the mechanism of trauma (fall versus kick), rider demographics, equestrian experience, protective equipment, and whether or not a horse was shod on the anatomic site of a horse-related maxillofacial fracture, operating time, postoperative complication rate, and length of hospital stay. Methods: We retrospectively reviewed the medical records of patients treated for horse-related maxillofacial fractures at a single institution in Germany between January 2000 and March 2015. We used linear and logistic regression to test the above-mentioned...
Assessment of Vein Diameter and Body Surface Temperature after High-Intensity Laser Therapy (HILT) on the Tarsal Joint in Healthy Horses.
Journal of equine veterinary science    July 22, 2020   Volume 93 103198 doi: 10.1016/j.jevs.2020.103198
Godlewska M, Soroko M, Zielińska P.The aim of the study was to assess the influence of high-intensity laser therapy (HILT) on vein diameter and on body surface temperature changes in the tarsal joint area in a group of clinically healthy racehorses. The hypothesis of the study was that HILT increases vein diameter and body surface temperature in healthy tissue. The study involved 16 Thoroughbreds being subjected to ultrasonographic examination to assess changes of diameter of the cranial branch of the medial saphenous vein and thermographic examination to indicate temperature change of the dorsal surface of the tarsal joint, ju...
Publisher Correction: Differences in the intrinsic chondrogenic potential of equine umbilical cord matrix and cord blood mesenchymal stromal/stem cells for cartilage regeneration.
Scientific reports    July 21, 2020   Volume 10, Issue 1 12075 doi: 10.1038/s41598-020-69170-0
Rakic R, Bourdon B, Demoor M, Maddens S, Saulnier N, Galéra P.An amendment to this paper has been published and can be accessed via a link at the top of the paper.
Fluoroquinolone exposure in utero did not affect articular cartilage of resulting foals.
Equine veterinary journal    July 20, 2020   Volume 53, Issue 2 385-396 doi: 10.1111/evj.13295
Ellerbrock RE, Canisso IF, Larsen RJ, Garrett KS, Stewart MC, Herzog KK, Kersh ME, Moshage SG, Podico G, Lima FS, Childs BA.Recent studies have shown that fluoroquinolones, specifically, enrofloxacin and its active metabolite (ciprofloxacin), cross the equine placenta without causing gross or histological lesions in the first and third trimester fetuses or resulting foal. However, it is possible that in utero exposure to fluoroquinolones may cause subtle lesions not detectable by standard means; thus, a more in-depth assessment of potential toxicity is warranted. Objective: To use quantitative magnetic resonance imaging (qMRI), biomechanical testing, and chondrocyte gene expression to evaluate the limbs of foals ex...
Period of excretion of equine herpesvirus 3 (EHV-3) from a stallion before showing clinical signs of equine coital exanthema and the effect of acyclovir treatment on the duration of EHV-3 excretion.
The Journal of veterinary medical science    July 20, 2020   Volume 82, Issue 9 1299-1305 doi: 10.1292/jvms.20-0056
Toishi Y, Tsunoda N, Kirisawa R.In 2017, two Thoroughbred stallions, A and B in Farms A and B, respectively, in Hokkaido in Japan showed clinical signs of equine coital exanthema (ECE). In 2020, stallion C in Farm B showed clinical signs of ECE. Eighteen mares were mated within five days before stallion A developed ECE. Ten mares that mated within 3 days before onset showed clinical signs of ECE on the external genitalia. Equine herpesvirus 3 (EHV-3) was isolated from vaginal swabs from three mares that mated within 2 days before onset. Swabs from 12 mares that mated within 4 days before onset were real-time PCR (rPCR)-posit...
Use of a chronic soft tissue expansion device to facilitate blepharoplasty in a horse with lower-lid cicatricial ectropion with a 14-year follow-up.
Veterinary ophthalmology    July 20, 2020   Volume 23, Issue 5 899-904 doi: 10.1111/vop.12806
Whittaker CJ, Reynolds BD, McCarthy PM, Taylor SF, Major D, Caruso KA, Smith J.A 3-year-old Thoroughbred filly presented to a referral equine hospital for surgical correction of a severe cicatricial lateral lower eyelid ectropion OD, with secondary exposure keratitis. The severity of the ectropion deemed that conventional ectropion repairs would be unsuccessful. Therefore, a soft tissue expansion device was used to create sufficient local tissue for a rotational graft with tension-relieving horizontal incisions to be performed to facilitate closure and acceptable eyelid apposition. The keratitis had resolved by two months postoperatively. There were no long-term complica...
Variation in the MRI signal intensity of naturally occurring equine superficial digital flexor tendinopathies over a 12-month period.
The Veterinary record    July 20, 2020   Volume 187, Issue 7 e53 doi: 10.1136/vr.105800
Berner D, Brehm W, Gerlach K, Offhaus J, Scharner D, Burk J.Signal intensity (SI) of equine tendinopathies in MRI differs between the superficial digital flexor tendon (SDFT) and the deep digital flexor tendon (DDFT). In DDFT lesions, short tau inversion recovery (STIR) SI decreases earlier than T2-weighted (T2w) SI, while the latter decreases earlier in SDFT lesions, but long-term results using STIR sequences are lacking. Methods: Standing MRI of eight horses with naturally occurring SDFT lesions was performed at the day of treatment as well as 2, 6 and 12 months after treatment. Results: After 12 months, six horses were sound and showed complete reso...
Arthroscopic debridement of short frontal plane proximal phalanx fractures preserves racing performance.
Equine veterinary journal    July 18, 2020   Volume 53, Issue 2 267-276 doi: 10.1111/evj.13275
Elzer EJ, Bramlage LR.Outcomes have been reported for a limited number of short frontal plane fractures of the proximal phalanx following nonsurgical treatment and internal fixation. Objective: To describe a new approach, arthroscopic debridement, of short frontal plane fractures of the proximal phalanx in flat-racing Thoroughbreds and post-operative racing outcome. Methods: Retrospective case-control study. Methods: Medical records of 81 Thoroughbred racehorses treated with arthroscopic debridement for frontal plane fractures of the proximal phalanx were reviewed. Diagnostic images and operative reports were used ...
Clinical use of computer-assisted orthopedic surgery in horses.
Veterinary surgery : VS    July 17, 2020   Volume 49, Issue 6 1075-1087 doi: 10.1111/vsu.13486
de Preux M, Klopfenstein Bregger MD, Brünisholz HP, Van der Vekens E, Schweizer-Gorgas D, Koch C.To describe clinical applications of computer-assisted orthopedic surgery (CAOS) in horses with a navigation system coupled with a cone beam computed tomography unit. Methods: Retrospective clinical case series. Methods: Thirteen adult horses surgically treated with CAOS. Methods: Medical records were searched for horses that underwent CAOS between 2016 and 2019. Data retrieved included signalment, diagnosis, lameness grade prior to surgery, surgical technique and complications, anesthesia and surgery time, and information pertaining to the perioperative case management and outcome. Results: I...
Puncture of the Equine Embryonic Capsule and Its Repair In Vivo and In Vitro.
Journal of equine veterinary science    July 17, 2020   Volume 93 103194 doi: 10.1016/j.jevs.2020.103194
Wilsher S, Rigali F, Kovacsy S, Allen WT.Vitrification of embryos >300 µm in diameter requires puncture of the glycoprotein capsule, although the size of the hole compatible with embryo survival is unknown. Forty-five day-7 or -8 embryos were punctured using a 30-µm glass biopsy pipette mounted on a micromanipulator (n = 20) or manually with either an acupuncture needle (∼100-µm diameter -hole; n = 10) or a microneedle with a <1 µm tip to produce a ∼30-µm diameter hole (n = 15) before transferring to recipient mares; further 12 embryos were punctured with either the acupuncture needle or microneedle before being cultured ...