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Topic:Veterinary Medicine

Veterinary medicine for horses encompasses the study and application of medical practices to diagnose, treat, and prevent diseases in equine species. This field involves a comprehensive understanding of equine anatomy, physiology, pathology, and pharmacology. Veterinary practitioners employ a range of diagnostic tools and therapeutic interventions to address health issues in horses, including lameness, gastrointestinal disorders, respiratory conditions, and infectious diseases. Preventative care, such as vaccination and deworming programs, is also a significant aspect of equine veterinary medicine. This page gathers peer-reviewed research studies and scholarly articles that explore various aspects of veterinary medicine as it pertains to horses, including advancements in diagnostic techniques, treatment protocols, and preventive health strategies.
No Evidence of Mosquito Involvement in the Transmission of Equine Hepacivirus (Flaviviridae) in an Epidemiological Survey of Austrian Horses.
Viruses    November 1, 2019   Volume 11, Issue 11 1014 doi: 10.3390/v11111014
Badenhorst M, de Heus P, Auer A, Rümenapf T, Tegtmeyer B, Kolodziejek J, Nowotny N, Steinmann E, Cavalleri JV.Prevalence studies have demonstrated a global distribution of equine hepacivirus (EqHV), a member of the family Flaviviridae. However, apart from a single case of vertical transmission, natural routes of EqHV transmission remain elusive. Many known flaviviruses are horizontally transmitted between hematophagous arthropods and vertebrate hosts. This study represents the first investigation of potential EqHV transmission by mosquitoes. More than 5000 mosquitoes were collected across Austria and analyzed for EqHV ribonucleic acid (RNA) by reverse transcription quantitative polymerase chain reacti...
Effects of media and promoters on different lipid peroxidation assays in stallion sperm.
Animal reproduction science    November 1, 2019   Volume 211 106199 doi: 10.1016/j.anireprosci.2019.106199
Ghosh S, Serafini R, Love CC, Teague SR, Hernández-Avilés C, LaCaze KA, Varner DD.Effects of different media and promoters on lipid peroxidation (LPO) in viable stallion sperm have not been reported. Aims of this study were to determine effects of three media (INRA-96™, Equipro CoolGuard™, and Biggers, Whitten and Whittingham [BWW]), and promoters (iron sulfate-Fe; ultraviolet light-UV; or control-no exposure to promoters) on viable sperm LPO using four different flow cytometric assays (i.e., BODIPY, Liperfluo, 4-hydroxylnonenal [4HNE], malonaldehyde [MDA]). Significant media x promoter interactions were detected using the Liperfluo, 4HNE, and MDA assays (P <  0.05)...
Relaxing effects of clenbuterol, ritodrine, salbutamol and fenoterol on the contractions of horse isolated bronchi induced by different stimuli.
Research in veterinary science    October 31, 2019   Volume 128 43-48 doi: 10.1016/j.rvsc.2019.10.022
Pozzoli C, Bertini S, Poli E, Placenza G, Menozzi A.β-adrenoceptor agonists are considered the most effective drugs to counteract bronchoconstriction in horses with asthma, but only clenbuterol is commonly employed in clinical practice. We evaluated the effects of different selective β agonists: clenbuterol, ritodrine, salbutamol, and fenoterol on the contractions of isolated bronchial muscle of horses induced by electrical field stimulation (EFS), carbachol, histamine, and KCl. All β agonists reduced the amplitude of contraction induced by the different stimuli but with variable efficacy and potency. Fenoterol and salbutamol were more effec...
Bleaching melanin in formalin-fixed and paraffin-embedded melanoma specimens using visible light: a pilot study.
European journal of histochemistry : EJH    October 31, 2019   Volume 63, Issue 4 doi: 10.4081/ejh.2019.3071
Pigoli C, Gibelli LR, Caniatti M, Moretti L, Sironi G, Giudice C.In fluorescence microscopy, light radiation can be used to bleach fluorescent molecules in formalin-fixed and paraffin-embedded (FFPE) samples, in order to increase the ratio between signal of interest and background autofluorescence. We tested if the same principle can be exploited in bright field microscopy to bleach pigmented melanoma FFPE sections together with cell morphology maintenance. After dewaxing and rehydration, serial FFPE sections of a feline diffuse iris melanoma, a canine dermal melanoma, a gray horse dermal melanoma and a swine cutaneous melanoma were irradiated with visible ...
Cutaneous Leishmaniosis caused by Leishmania martiniquensis in a Horse in Florida.
Journal of comparative pathology    October 31, 2019   Volume 173 13-18 doi: 10.1016/j.jcpa.2019.09.011
Menezes RC, Campos MP, Popielarczyk M, Kiupel M.We report a new case of cutaneous leishmaniosis caused by Leishmania (Mundinia) martiniquensis in a horse in Florida, USA. A 10-year-old neutered male Quarter horse was presented with multifocal to coalescing, raised, ulcerated and oozing, non-healing wounds on both pinnae of several weeks' duration. After a few months, the lesions regressed spontaneously. Biopsies of the lesions were performed with microscopical findings of epidermal hyperplasia with multifocal ulceration and focally extensive, dermal pyogranulomatous inflammation with numerous intact and degenerate neutrophils being surround...
Inter- and Intra-Rater Reliability of Soft Tissue Palpation Scoring in the Equine Thoracic Epaxial Region.
Journal of equine veterinary science    October 31, 2019   Volume 83 102812 doi: 10.1016/j.jevs.2019.102812
Merrifield-Jones M, Tabor G, Williams J.Back pain is a significant factor for horses and is challenging for professionals to diagnose, with assessment frequently using subjective tools such as manual palpation. Reliable and valid objective measures are required and use of a pressure algometer (PA) has been investigated as an assessment tool; however, it has limitations, and other more realistic methods may be better suited for the task. The aim of the study was to establish inter- and intra-rater reliability for PA, FlexiForce Sensor (FFS), and manual palpation for equine epaxial soft tissue, measuring mechanical nociception thresho...
Assessment of Leishmania infantum infection in equine populations in a canine visceral leishmaniosis transmission area.
BMC veterinary research    October 30, 2019   Volume 15, Issue 1 381 doi: 10.1186/s12917-019-2108-1
Escobar TA, Dowich G, Dos Santos TP, Zuravski L, Duarte CA, Lübeck I, Manfredini V.Leishmaniosis, zoonosis that produces significant public health impacts, is caused by Leishmania infantum. Canines are the main domestic reservoir and, besides humans, other species of mammals could be infected when living in endemic areas. In this study, we detected equine Leishmania infantum infections in a canine visceral leishmaniosis transmission area and evaluated the clinical, haematological, biochemical and oxidative stress disorders. This study was conducted in Uruguaiana, Rio Grande do Sul, south of Brazil. Peripheral blood samples were collected from 124 animals (98 horses and 26 do...
Ex vivo evaluation of an alternative technique for resection of the proximal manica flexoria in horses.
Veterinary surgery : VS    October 30, 2019   Volume 49, Issue 2 401-408 doi: 10.1111/vsu.13336
Noguera Cender A, Mählman K, Lischer CJ.To describe and evaluate an alternative technique for tenoscopic resection of the manica flexoria (MF). Methods: Descriptive study. Methods: Equine cadaver forelimbs and hind limbs (n = 36). Methods: Three surgeons varying in experience resected the MF in 12 limbs. The procedure was divided into six steps, including resection of the lateral/medial borders of the MF with a hook knife and resection of the proximal border with a micro scalpel. Outcomes compared between surgeons included completeness of resection, appearance of the margins of resection, appearance of the resected MF, collateral da...
Autologous bone marrow mononuclear cells modulate joint homeostasis in an equine in vivo model of synovitis.
FASEB journal : official publication of the Federation of American Societies for Experimental Biology    October 30, 2019   Volume 33, Issue 12 14337-14353 doi: 10.1096/fj.201901684RR
Menarim BC, Gillis KH, Oliver A, Mason C, Ngo Y, Werre SR, Barrett SH, Luo X, Byron CR, Dahlgren LA.Osteoarthritis (OA) is characterized by macrophage-driven synovitis. Macrophages promote synovial health but become inflammatory when their regulatory functions are overwhelmed. Bone marrow mononuclear cells (BMNCs) are a rich source of macrophage progenitors used for treating chronic inflammation and produce essential molecules for cartilage metabolism. This study investigated the response to autologous BMNC injection in normal and inflamed joints. Synovitis was induced in both radiocarpal joints of 6 horses. After 8 h, 1 inflamed radiocarpal and 1 normal tarsocrural joint received BMNC injec...
Estimation of glomerular filtration rate by plasma clearance of iohexol in healthy horses of various ages.
Journal of veterinary internal medicine    October 30, 2019   Volume 33, Issue 6 2765-2769 doi: 10.1111/jvim.15642
Plasma clearance of iohexol is a reliable method to assess the glomerular filtration rate (GFR). The relationship between aging and GFR in horses is unclear. To compare GFR estimated by iohexol clearance in horses of different ages. Twenty-one clinically healthy horses were enrolled. Prospective study. Groups: (A) composed by 8 young horses (≤14 years old) with serum creatinine <1.5 mg/dL, (B) by 7 aged horses (≥20 years old) with serum creatinine <1.5 mg/dL, and (C) by 6 aged horses (≥20 years old) with serum creatinine ≥1.5 mg/dL. Iohexol was injected (75.5 mg/kg) through...
Exercise Induced Laryngeal Obstruction in Humans and Equines. A Comparative Review.
Frontiers in physiology    October 30, 2019   Volume 10 1333 doi: 10.3389/fphys.2019.01333
Fretheim-Kelly ZL, Halvorsen T, Clemm H, Roksund O, Heimdal JH, Vollsæter M, Fintl C, Strand E.Dynamic obstructions of the larynx are a set of disorders that occur during exercise in equines and humans. There are a number of similarities in presentation, diagnosis, pathophysiology and treatment. Both equines and humans present with exercise intolerance secondary to dyspnea. During laryngoscopy at rest, the larynx appears to function normally. Abnormalities are only revealed during laryngoscopy at exercise, seemingly triggered by increased ventilatory demands, and quickly resolve after cessation of exercise. Lower airway disease (asthma being the most prevalent condition), cardiac diseas...
The mule (Equus mulus) as a recipient of horse (Equus caballus) embryos: Comparative aspects of early pregnancy with mares.
Theriogenology    October 30, 2019   Volume 145 217-225 doi: 10.1016/j.theriogenology.2019.10.029
Camargo CE, Rechsteiner SF, Macan RC, Kozicki LE, Gastal MO, Gastal EL.The aim of this study was to compare the embryonic and early fetal development of horse embryos between recipient mules and mares from day 10-60 of pregnancy, in addition to hormonal (eCG and progesterone), ovarian, and uterine characteristics for approximately 4 months. Embryo donor mares (n = 5) and two groups of recipients (acyclic mules, n = 7; cyclic mares, n = 7) were used. Donor mares were monitored daily by transrectal ultrasonography and inseminated using fresh semen. Cyclic recipient mares were synchronized with the donor's ovulation using PGF2α and deslorelin acetate. M...
Enteroinsular axis response to carbohydrates and fasting in healthy newborn foals.
Journal of veterinary internal medicine    October 30, 2019   Volume 33, Issue 6 2752-2764 doi: 10.1111/jvim.15641
Rings LM, Swink JM, Dunbar LK, Burns TA, Toribio RE.The enteroinsular axis (EIA) comprises intestinal factors (incretins) that stimulate insulin release after PO ingestion of nutrients. Glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) are the main incretins. The EIA has not been investigated in healthy neonatal foals but should be important because energy demands are high in healthy foals and dysregulation is frequent in sick foals. Objective: To evaluate the EIA response to carbohydrates or fasting in newborn foals. We hypothesized that incretin secretion would be higher after PO versus IV carbohydrate adm...
An Equine Herpesvirus Type 1 (EHV-1) Ab4 Open Reading Frame 2 Deletion Mutant Provides Immunity and Protection from EHV-1 Infection and Disease.
Journal of virology    October 29, 2019   Volume 93, Issue 22 e01011-19 doi: 10.1128/JVI.01011-19
Schnabel CL, Babasyan S, Rollins A, Freer H, Wimer CL, Perkins GA, Raza F, Osterrieder N, Wagner B.Equine herpesvirus type 1 (EHV-1) outbreaks continue to occur despite widely used vaccination. Therefore, development of EHV-1 vaccines providing improved immunity and protection is ongoing. Here, an open reading frame 2 deletion mutant of the neuropathogenic EHV-1 strain Ab4 (Ab4ΔORF2) was tested as a vaccine candidate. Three groups of horses ( = 8 each) were infected intranasally with Ab4ΔORF2 or the parent Ab4 virus or were kept as noninfected controls. Horses infected with Ab4ΔORF2 had reduced fever and nasal virus shedding compared to those infected with Ab4 but mounted similar ada...
Superovulation, embryo recovery, and pregnancy rates from seasonally anovulatory donor mares treated with recombinant equine FSH (reFSH).
Theriogenology    October 29, 2019   Volume 142 291-295 doi: 10.1016/j.theriogenology.2019.10.030
Roser JF, Etcharren MV, Miragaya MH, Mutto A, Colgin M, Losinno L, Ross PJ.The effectiveness of different treatments with recombinant equine FSH to stimulate follicular growth, multiple ovulations and embryo production in seasonally anovulatory mares was evaluated. During mid-winter season (July-August in Argentina, South America) forty light breed donor mares, presenting follicles <10 mm in diameter and no CL at ultrasound examination (deep-anestrus), were randomly assigned (n = 10/group) to one of the following treatments: Group 1: twice daily intramuscular (IM) injections of 0.65 mg reFSH (AspenBio Pharma, CO), Group 2: once daily IM injection of 1.3 mg ...
Impact of alpha-tocopherol deficiency and supplementation on sacrocaudalis and gluteal muscle fiber histopathology and morphology in horses.
Journal of veterinary internal medicine    October 29, 2019   Volume 33, Issue 6 2770-2779 doi: 10.1111/jvim.15643
Bookbinder L, Finno CJ, Firshman AM, Katzman SA, Burns E, Peterson J, Dahlgren A, Ming-Whitfield B, Glessner S, Borer-Matsui A, Valberg SJ.A subset of horses deficient in alpha-tocopherol (α-TP) develop muscle atrophy and vitamin E-responsive myopathy (VEM) characterized by mitochondrial alterations in the sacrocaudalis dorsalis medialis muscle (SC). Objective: To quantify muscle histopathologic abnormalities in subclinical α-TP deficient horses before and after α-TP supplementation and compare with retrospective (r)VEM cases. Methods: Prospective study; 16 healthy α-TP-deficient Quarter Horses. Retrospective study; 10 retrospective vitamin E-responsive myopathy (rVEM) cases . Methods: Blood, SC, and gluteus medius (GM) biops...
African horse sickness: a potential threat to the UK.
The Veterinary record    October 28, 2019   Volume 185, Issue 16 501-503 doi: 10.1136/vr.l6189
The UK has never experienced an outbreak of African horse sickness but, as Matthew Robin of the Fyrnwy Equine Clinic explains, there is still a risk of a case occurring.
Nonsurgical and surgical management of metacarpo/metatarsophalangeal joint dorsal chip fracture in the Thoroughbred racehorse.
Equine veterinary journal    October 28, 2019   Volume 52, Issue 3 399-403 doi: 10.1111/evj.13185
Ramzan PHL, Wylie CE.Arthroscopy is considered the treatment of choice for dorsal osteochondral chip fractures of the metacarpo/metatarsophalangeal (MCP/MTP) joints in the racehorse; however, there is no published research on non-surgical management of this injury. Objective: To compare clinical features, intra-articular medication use and return to racing in Thoroughbred racehorses with non-surgically (non-SX) or surgically (SX) managed MCP/MTP dorsal chip fracture. Methods: Retrospective observational study conducted between 2006 and 2014. Methods: Radiographs of Thoroughbred racehorses were reviewed to identify...
Application of embryo biopsy and sex determination via polymerase chain reaction in a commercial equine embryo transfer program in Argentina.
Reproduction, fertility, and development    October 28, 2019   Volume 31, Issue 12 1917-1925 doi: 10.1071/RD19228
Riera FL, Roldán JE, Espinosa JM, Fernandez JE, Ortiz I, Hinrichs K.Embryo biopsy for fetal sexing has clinical application, but few reports are available of its use within an active embryo transfer program. We evaluated results on biopsy of 459 embryos over one breeding season. There were no significant differences in pregnancy rate between biopsied and non-biopsied embryos (72% vs 73%) or for biopsied embryos recovered at the centre (73%) compared with those shipped overnight (72%). However, the pregnancy rate decreased significantly in shipped embryos biopsied ≥20h after collection. Overall, 86% of biopsies provided a sex diagnosis. The likelihood of a po...
BEVA primary care clinical guidelines: Analgesia.
Equine veterinary journal    October 28, 2019   Volume 52, Issue 1 13-27 doi: 10.1111/evj.13198
Bowen IM, Redpath A, Dugdale A, Burford JH, Lloyd D, Watson T, Hallowell GD.Primary care guidelines provide a reference point to guide clinicians based on a systematic review of the literature, contextualised by expert clinical opinion. These guidelines develop a modification of the GRADE framework for assessment of research evidence (vetGRADE) and applied this to a range of clinical scenarios regarding use of analgesic agents. Key guidelines produced by the panel included recommendations that horses undergoing routine castration should receive intratesticular local anaesthesia irrespective of methods adopted and that horses should receive NSAIDs prior to surgery (ove...
Colic surgery: keeping it affordable for horse owners.
The Veterinary record    October 28, 2019   Volume 185, Issue 16 505-507 doi: 10.1136/vr.l6062
Archer DC.No abstract available
Re-evaluation of the pharmacokinetics of xylazine administered to Thoroughbred horses.
Journal of veterinary pharmacology and therapeutics    October 25, 2019   Volume 43, Issue 1 6-12 doi: 10.1111/jvp.12820
Habershon-Butcher J, Cutler C, Viljanto M, Hincks PR, Biddle S, Paine SW.Xylazine is widely used worldwide as a short-acting sedative in general equine and racing practice. In the UK, although it has a legitimate use during training, equine anti-doping rules state it is a prohibited substance on race day. The aim of the study was to produce a detection time (DT) to better inform European veterinary surgeons so that xylazine can be used appropriately under regulatory rules. Previous publications have various limitations pertaining to analysis method, particularly for plasma and limited length of time of sample collection. In this study, pharmacokinetic data were pro...
Investigation into pathophysiology of naturally occurring palatal instability and intermittent dorsal displacement of the soft palate (DDSP) in racehorses: Thyro-hyoid muscles fatigue during exercise.
PloS one    October 25, 2019   Volume 14, Issue 10 e0224524 doi: 10.1371/journal.pone.0224524
Cercone M, Olsen E, Perkins JD, Cheetham J, Mitchell LM, Ducharme NG.Exercise induced intermittent dorsal displacement of the soft palate (DDSP) is a common cause of airway obstruction and poor performance in racehorses. The definite etiology is still unclear, but through an experimental model, a role in the development of this condition was identified in the dysfunction of the thyro-hyoid muscles. The present study aimed to elucidate the nature of this dysfunction by investigating the spontaneous response to exercise of the thyro-hyoid muscles in racehorses with naturally occurring DDSP. Intramuscular electrodes were implanted in the thyro-hyoid muscles of nin...
Influence of digital hypothermia on lamellar events related to IL-6/gp130 signalling in equine sepsis-related laminitis.
Equine veterinary journal    October 24, 2019   Volume 52, Issue 3 441-448 doi: 10.1111/evj.13184
Dern K, Burns TA, Watts MR, van Eps AW, Belknap JK.Interleukin-6 (IL-6) is consistently increased in the digital lamellae in different studies of sepsis-related laminitis (SRL). IL-6 signalling through the gp130 receptor activates similar signalling (i.e. mTORC1-related signalling) previously reported to be activated in models of endocrinopathic laminitis. Objective: To assess the activation state of signalling proteins downstream of IL-6/gp130 receptor complex activation in an experimental model of SRL. Methods: Randomised experimental study. Methods: Lamellar phospho-(P) protein concentrations downstream of the IL-6/gp130 receptors were asse...
Use of rotational periosteal flaps in primary repair of sinofacial fractures in seven horses.
New Zealand veterinary journal    October 24, 2019   Volume 68, Issue 2 119-125 doi: 10.1080/00480169.2019.1675555
Gustafsson K, Tatz AJ, Dahan R, Kelmer G.Case history: Medical records were reviewed of horses (n = 7) undergoing surgery for fracture of one or more facial bones extending into the paranasal sinuses that was repaired primarily within 24 hours of the time of injury using a rotational periosteal flap, between April 2009 and May 2017. A kick from another horse was the cause of the injury of three horses, and one horse was injured when it collided with a tree. The cause of the injury of three horses was unknown.Clinical findings and treatment: Fractures were of the right maxillary bone in two horses, the left maxillary bone in two h...
Atrial premature depolarisations five days post electrical cardioversion are related to atrial fibrillation recurrence risk in horses.
Equine veterinary journal    October 24, 2019   Volume 52, Issue 3 374-378 doi: 10.1111/evj.13186
Vernemmen I, De Clercq D, Decloedt A, Vera L, Van Steenkiste G, van Loon G.The number of atrial premature depolarisations (APDs) is a known risk factor for atrial fibrillation (AF) recurrence in humans. Objective: To evaluate if the number of APDs over a 24-h period 5 days post cardioversion predicts AF recurrence within 1 year in horses, taking the multifactorial nature of AF into account. Methods: Retrospective case series. Methods: Eighty horses met these inclusion criteria: first AF episode, no AF recurrence within 5 days post cardioversion, cardioversion by transvenous electrical cardioversion (TVEC), 24-h ECG recording and echocardiographic examination 5 da...
Single-dose pharmacokinetics of orally and rectally administered misoprostol in adult horses.
American journal of veterinary research    October 24, 2019   Volume 80, Issue 11 1026-1033 doi: 10.2460/ajvr.80.11.1026
Lopp CT, McCoy AM, Boothe D, Schaeffer DJ, Lascola K.To characterize the pharmacokinetics of a clinically relevant dose of misoprostol administered PO or per rectum (PR) to horses. Methods: 8 healthy adult horses. Methods: In a randomized 3-way crossover design, horses received a single dose of misoprostol (5 μg/kg) administered PO (with horses fed and unfed) and PR, with a minimum 3-week washout period separating the experimental conditions. Blood samples were obtained before and at various points after drug administration (total, 24 hours), and plasma concentrations of misoprostol free acid were measured. Results: Mean maximum plasma concentr...
Update on advanced semen-processing technologies and their application for in vitro embryo production in horses.
Reproduction, fertility, and development    October 24, 2019   Volume 31, Issue 12 1771-1777 doi: 10.1071/RD19301
Morris LH, Maclellan LJ.The increased commercialisation of intracytoplasmic sperm injection (ICSI) in horses creates more opportunities to incorporate advanced reproductive technologies, such as sex-sorted, refrozen and lyophilised spermatozoa, into a breeding program. This paper reviews the status of these semen-handling technologies in light of their use in equine ICSI programs. Pregnancies have been achieved from each of these advanced technologies when combined with ICSI in horses, but refinements in the semen-handling processes underpinning these technologies are currently being explored to produce more reliable...
Plasma adrenocorticotropic hormone concentration in horses decreases after freezing for 60 days. Haffner JC, Neal DL, Hoffman RM, Grubbs ST.We investigated the stability of adrenocorticotropic hormone (ACTH) in plasma after freezing for different lengths of time. The plasma ACTH concentrations of 12 horses were measured on day 0 (baseline) and over time, after stimulation with thyrotropin-releasing hormone. Samples were stored at -80°C for 3, 7, 30, 60, and 90 d, or at -20°C for 3, 7, 30, and 60 d, or between ice packs at -20°C for 3 and 7 d prior to determination of ACTH concentration. ACTH concentrations were compared to baseline (non-frozen day 0 plasma) for each storage method using a mixed model with repeated measure...
Effect of hCG application at different moments of the estrous cycle on corpus luteum and uterine vascularization and serum progesterone concentration in mares.
Animal reproduction    October 24, 2019   Volume 16, Issue 2 317-327 doi: 10.21451/1984-3143-AR2018-0103
Alonso MA, Silva LA, Affonso FJ, Lemes KM, Celeghini ECC, Lançoni R, Carvalho HF, de Arruda RP.Establishment of pregnancy after embryo transfer is the ultimate goal of an embryo transfer program and increasing pregnancy rates and reducing pregnancy loss are mandatory. The utilization of treatments to improve conception rates in recipient mares has been the focus of several research groups over the last years and the results are controversial. Some studies using human chorionic gonadotrophin (hCG) found promising results. Our hypothesis was that hCG administration would cause an additional stimulation on luteal function, uterine and luteal vascularization and progesterone concentration, ...