Medicine and Health Sciences Commons^™

Equine Fecal Microbiota Response To Short Term Antibiotic Administration, Jordan Lacey Parker

Theses and Dissertations--Veterinary Science

Though generally regarded as safe, research continues to demonstrate negative side effects of antibiotic administration on the gastrointestinal (GIT) microbiota across species. In horses, antibiotic associated diarrhea (AAD) is a life-threatening side effect linked to the GIT microbiota. This study tested the hypothesis that short term antibiotic administration to healthy horses would negatively impact the fecal microbiota as measured by their ability to digest nutrients and produce short chain fatty acids (SCFAs). Twenty-four horses were assigned to one of four treatments: control (CO); potassium penicillin/gentamicin sulfate (KPG); ceftiofur crystalline free acid (EX); trimethoprim/sulfamethoxazole (SMZ); and treated for 4 days. Fecal …

Identifying Bisphosphonate Protein Biomarkers In Equine Sera Using Mass Spectrometry Methods, Malinda Jayne Alison Porter

Theses and Dissertations--Veterinary Science

Bone resorptive diseases affect humans and horses, alike. Examples of these diseases include osteoporosis, Paget’s disease, and osteogenesis imperfecta in humans, and navicular disease in the horse (Mitchell et al., 2019; Suva et al., 2021). Bisphosphonates are used to treat these diseases, and may remain in the bone for several years after administration, demonstrating the need for newer drug testing methods.

The purpose of the discovery phase of this study was to use blood samples of in-training horses which had been given an initial administration of the bisphosphonate tiludronate and identify protein biomarkers that changed in response to this class …

The Effects Of Cannabidiol On Senior Equine Health, Shelley Turner

Theses and Dissertations--Veterinary Science

Cannabis sativa L., has been revealed to produce hundreds of phytocannabinoids, of which cannabidiol (CBD) is one of the most desired. It has been revealed that CBD can potentially treat inflammation and act as an analgesic in humans without psychoactive effects of delta-9-tetrahydrocannabinol (THC). Recently, there has been interest in understanding the potential health benefits of CBD for horses. With an increasing senior horse population (< 15 years old), alternatives to the use of non-steroidal anti-inflammatory drugs (NSAIDs) such as phenylbutazone, firocoxib, and flunixin meglumine, are desired as these common anti-inflammatory treatments have negative side effects. Because senior horses may have chronic low-grade inflammation, termed inflamm-aging, they are a natural model to investigate the efficacy of CBD on inflammatory responses as well as other health parameters. Given this interest, and due to the lack of research conducted on CBD in the horse, the overall objective of this dissertation work was to conduct both basic and applied research investigating the effects of CBD on equine immune function and health. The first objective was to determine the in vitro effects of CBD as an anti-inflammatory. Peripheral blood mononuclear cells (PBMCs) from senior horses were cultured in vitro with increasing concentrations of pure CBD dissolved in dimethyl sulfoxide (DMSO). These cells were …

The Effect Of Intra-Articular Corticosteroids On The Systemic Messenger Rna Response In An Equine Experimental Inflammation Model, Emma Elaine Partridge

Theses and Dissertations--Veterinary Science

Intra-articular (IA) corticosteroids are regularly used in equine athletes for the control of joint inflammation. The goal of this study was to utilize an acute synovitis inflammation model to determine the residual effects of IA betamethasone and triamcinolone acetonide on various inflammatory parameters and lameness.

Five mixed-breed, 2-year-old horses were randomly allocated to an IA treatment of the radiocarpal joint with 9 mg of either betamethasone or triamcinolone acetonide. Two weeks following treatment, horses were injected with 1μg of lipopolysaccharide (LPS) diluted in 1 mL of 0.9% sterile saline. Following LPS injection, horses were crossed-over and both sets of injections …

Estrogens Regulate Placental Angiogenesis In Horses, Shingo Haneda, Pouya Dini, Alejandro Esteller-Vico, Kirsten E. Scoggin, Edward L. Squires, Mats H. T. Troedsson, Peter Daels, Yasuo Nambo, Barry A. Ball

Maxwell H. Gluck Equine Research Center Faculty Publications

A sufficient vascular network within the feto-maternal interface is necessary for placental function. Several pregnancy abnormalities have been associated with abnormal vascular formations in the placenta. We hypothesized that growth and expansion of the placental vascular network in the equine (Equus caballus) placenta is regulated by estrogens (estrogen family hormones), a hormone with a high circulating concentration during equine gestation. Administration of letrozole, a potent and specific inhibitor of aromatase, during the first trimester (D30 to D118), decreased circulatory estrone sulfate concentrations, increased circulatory testosterone and androstenedione concentrations, and tended to reduce the weight of the fetus ( …

Transcriptomic Analysis Of Equine Chorioallantois Reveals Immune Networks And Molecular Mechanisms Involved In Nocardioform Placentitis, Hossam El-Sheikh Ali, Shavahn C. Loux, Laura Kennedy, Kirsten E. Scoggin, Pouya Dini, Carleigh E. Fedorka, Theodore S. Kalbfleisch, Alejandro Esteller-Vico, David W. Horohov, Erdal Erol, Craig N. Carter, Jackie L. Smith, Barry A. Ball

Maxwell H. Gluck Equine Research Center Faculty Publications

Nocardioform placentitis (NP) continues to result in episodic outbreaks of abortion and preterm birth in mares and remains a poorly understood disease. The objective of this study was to characterize the transcriptome of the chorioallantois (CA) of mares with NP. The CA were collected from mares with confirmed NP based upon histopathology, microbiological culture and PCR for Amycolatopsis spp. Samples were collected from the margin of the NP lesion (NPL, n = 4) and grossly normal region (NPN, n = 4). Additionally, CA samples were collected from normal postpartum mares (Control; CRL, n = 4). Transcriptome analysis identified 2892 differentially …

Intrauterine Blood Plasma Platelet-Therapy Mitigates Persistent Breeding-Induced Endometritis, Reduces Uterine Infections, And Improves Embryo Recovery In Mares, Lorenzo G. T. M. Segabinazzi, Igor F. Canisso, Giorgia Podico, Lais L. Cunha, Guilherme Novello, Michael F. Rosser, Shavanh C. Loux, Fabio S. Lima, Marco A. Alvarenga

Maxwell H. Gluck Equine Research Center Faculty Publications

Microorganisms, including pathogenic or opportunistic bacteria and fungi, may gain access to the uterus during breeding, and infectious endometritis plays a major role in equine subfertility. This study aimed to assess the post-breeding inflammatory response, endometrial culture, and embryo recovery of mares susceptible to persistent breeding-induced endometritis (PBIE) treated with plasma-rich (PRP) or -poor (PPP) plasma. Mares (n = 12) susceptible to PBIE had three cycles randomly assigned to receive intrauterine infusions of lactate ringer solution (LRS, control), or autologous PRP or PPP pre- (−48 and −24 h) and post-breeding (6 and 24 h). Mares were bred with fresh …

The Pelvic Flexure Separates Distinct Microbial Communities In The Equine Hindgut, Kailee J. Reed, Isabelle G. Z. Kunz, Jessica A. Scare, Martin K. Nielsen, Philip J. Turk, Robert J. Coleman, Stephen J. Coleman

Maxwell H. Gluck Equine Research Center Faculty Publications

As hindgut fermenters, horses are especially dependent on the microbiota residing in their cecum and large intestines. Interactions between these microbial populations and the horse are critical for maintaining gut homeostasis, which supports proper digestion. The current project was motivated to determine if any features of the fecal microbiota are informative of the microbial communities from the cecum, ventral colon, or dorsal colon. Digesta from the cecum, ventral colon, dorsal colon and feces were collected from 6 yearling miniature horses. Microbial DNA was isolated and the microbiota from each sample was characterized by profiling the V4 region of the 16S …

Stress Response After Race And Endurance Training Sessions And Competitions In Arabian Horses, O. Witkowska-Piłaszewicz, J. Grzędzicka, J. Seń, M. Czopowicz, M. Żmigrodzka, A. Winnicka, A. Cywińska, Craig N. Carter

Veterinary Diagnostic Laboratory Faculty Publications

Assuring a high level of animal welfare is a critical aspect of contemporary animal husbandry. Equine athletes begin their careers at a very young age when they are still developing and they are both physically and mentally immature. Lack of scientific knowledge of the stress related to horse racing impedes the development of optimal training programs to attain equilibrium between the best sport results and optimal welfare. This study aimed to determine the influence of the intensity and type of physical activity on peripheral blood cortisol concentration. Thirty untrained Arabians, 9 endurance and 21 race horses were enrolled in this …

Host-Parasite Interaction In Horses: Mucosal Responses To Naturally Acquired Cyathostomin Infections And Anthelmintic Treatment, Ashley Elaine Steuer

Theses and Dissertations--Veterinary Science

Cyathostomins are ubiquitous parasites in equids. In rare cases, cyathostomins lead to a generalized typhlocolitis and death. In healthy horses, local reactions are noted to the mucosal larvae; however, the mechanisms and importance of these reactions have not been elucidated. It has been hypothesized that anthelmintics can alter these reactions. Currently, three drug classes are approved for use in horses against cyathostomins; while all products target the adults, only two products are labeled as larvicidal. Adulticidal therapy is implicated in triggering the typhlocolitis, however, current evidence is contradictory. There is also conjecture that the larvicidal drugs can increase the risk …

Clinical Signs Associated With A Case Of Iatrogenic Poisoning By A Turpentine-Based Commercial Product In A 3-Day-Old Foal, Abelardo Morales Briceño, Hilal Al Mahrouqi, Abdallah Al Kayyoomi, Harmon Oliva, Kimberly Brewer, Thomas Tobin

Maxwell H. Gluck Equine Research Center Faculty Publications

The aim of this study is to report a case of iatrogenic poisoning by a turpentine-based commercial product in a 3-day-old foal and the successful treatment of this condition. A 3-day-old male foal presented with acute cardio-respiratory collapse, anaphylactic shock, severe respiratory distress, dyspnea and pulmonary edema. The animal’s history included previous administration of a commercial veterinary product (UNITIPAN). Clinical examination revealed: temperature: (39.9 °C), pulse: 135 bpm, respiration: 51 bpm, mucous membranes: moist and congested, capillary refill time: 3 sec or less. The foal was treated with emergency hydration, duphalyte, steroidal anti-inflammatory, atropine sulphate and antibiotic. After 6 h …

Effects Of Pituitary Pars Intermedia Dysfunction And Prascend® Treatment On Endocrine And Immune Function In Senior Horses, Ashton B. Miller

Theses and Dissertations--Veterinary Science

Pituitary pars intermedia dysfunction (PPID) is one of the most common endocrine diseases affecting senior horses. PPID causes abnormally high concentrations of adrenocorticotropic hormone (ACTH) in the plasma and a very distinct, long, shaggy haircoat (hypertrichosis). At present, the recommended treatment for PPID is daily oral administration of pergolide mesylate. Due to the increased ACTH levels associated with PPID, it is commonly thought that these horses are immunosuppressed and at increased risk of opportunistic infections, although current research in this area is sparse. Additionally, it is not well-understood how treatment with Prascend^® (pergolide tablets) affects endocrine measures other than …

An Epidemiologic Study Of Antimicrobial Resistance Of Staphylococcus Species Isolated From Equine Samples Submitted To A Diagnostic Laboratory, Ronita Adams, Jackie Smith, Stephan Locke, Erica Phillips, Erdal Erol, Craig N. Carter, Agricola Odoi

Veterinary Science Faculty Publications

Background

Antimicrobial resistance limits traditional treatment options and increases costs. It is therefore important to estimate the magnitude of the problem so as to provide empirical data to guide control efforts. The aim of this study was to investigate the burden and patterns of antimicrobial resistance (AMR) among equine Staphylococcus samples submitted to the University of Kentucky Veterinary Diagnostic Laboratory (UKVDL) from 1993 to 2009. Retrospective data of 1711 equine Staphylococcus samples submitted to the UKVDL during the time period 1993 to 2009 were included in the study. Antimicrobial susceptibility testing, that included 16 drugs, were performed using cultures followed …

Detection Of Strongylus Vulgaris In Equine Faecal Samples By Real-Time Pcr And Larval Culture – Method Comparison And Occurrence Assessment, A. Kaspar, K. Pfister, Martin K. Nielsen, C. Silaghi, H. Fink, M. C. Scheuerle

Veterinary Science Faculty Publications

Background: Strongylus vulgaris has become a rare parasite in Germany during the past 50 years due to the practice of frequent prophylactic anthelmintic therapy. To date, the emerging development of resistance in Cyathostominae and Parascaris spp. to numerous equine anthelmintics has changed deworming management and the frequency of anthelmintic usage. In this regard, reliable detection of parasitic infections, especially of the highly pathogenic S. vulgaris is essential. In the current study, two diagnostic methods for the detection of infections with S. vulgaris were compared and information on the occurrence of this parasite in German horses was gained. For this purpose, …

An Investigation Into Specific Seminal Plasma Proteins And Their Effect On The Innate Immune Response To Breeding In The Mare, Carleigh Elizabeth Fedorka

Theses and Dissertations--Veterinary Science

The mare experiences a transient innate immune response to breeding, the resolution of which is crucial for optimal fertility. The majority of mares are able to modulate this inflammation in a timely fashion, but a subpopulation exists which fail to do so and are considered susceptible to persistent breeding-induced endometritis (PBIE). Seminal plasma has been shown to modulate aspects of this inflammation. Recently, two seminal plasma proteins have garnered interest for their immune modulating properties: cysteine-rich secretory protein-3 (CRISP-3) and lactoferrin. These proteins have been found to alter the binding between sperm and neutrophils based on sperm viability in vitro …

Role Of Skeletal Paracrine Signals In The Proliferation And Chondrogenic Differentiation Of Interzone Cells, Parvathy Thampi

Theses and Dissertations--Veterinary Science

Articular cartilage in mammals has a limited intrinsic capacity to repair structural injuries and defects, a fact that contributes to the chronic and progressive nature of osteoarthritis. Current treatment modalities do not enable articular cartilage to achieve a complete and permanent restoration of normal structure and function with large or partial thickness lesions. In contrast to mammals, Mexican axolotl salamanders (Ambystoma mexicanum) have demonstrated the remarkable ability to spontaneously and completely repair even large joint cartilage lesions, an intrinsic healing process that involves interzone cells in the intraarticular space. Further, when interzone tissue is transplanted into critical sized …

Effect Of Dietary Starch Source And Concentration On Equine Fecal Microbiota, Brittany E. Harlow, Laurie M. Lawrence, Susan H. Hayes, Andrea Crum, Michael D. Flythe

Animal and Food Sciences Faculty Publications

Starch from corn is less susceptible to equine small intestinal digestion than starch from oats, and starch that reaches the hindgut can be utilized by the microbiota. The objective of the current study was to examine the effects of starch source on equine fecal microbiota. Thirty horses were assigned to treatments: control (hay only), HC (high corn), HO (high oats), LC (low corn), LO (low oats), and LW (low pelleted wheat middlings). Horses received an all-forage diet (2 wk; d -14 to d -1) before the treatment diets (2 wk; d 1 to 14). Starch was introduced gradually so that …

Annotation Of The Protein Coding Regions Of The Equine Genome, Matthew S. Hestand, Theodore S. Kalbfleisch, Stephen J. Coleman, Zheng Zeng, Jinze Liu, Ludovic Orlando, James N. Macleod

Veterinary Science Faculty Publications

Current gene annotation of the horse genome is largely derived from in silico predictions and cross-species alignments. Only a small number of genes are annotated based on equine EST and mRNA sequences. To expand the number of equine genes annotated from equine experimental evidence, we sequenced mRNA from a pool of forty-three different tissues. From these, we derived the structures of 68,594 transcripts. In addition, we identified 301,829 positions with SNPs or small indels within these transcripts relative to EquCab2. Interestingly, 780 variants extend the open reading frame of the transcript and appear to be small errors in the equine …

Diagnosis Of Strongylus Vulgaris, Martin K. Nielsen, Ulla Anderson, Daniel K. Howe

Veterinary Science Faculty Patents

Embodiments of the presently-disclosed subject matter provide an isolated polypeptide comprising the sequence of SEQ ID NO: 1, fragments thereof, and/or epitopes thereof. Embodiments of the presently-disclosed subject matter also provide methods for diagnosing a Strongylus vulgaris infection in a subject that comprises providing a biological sample from the subject and contacting the sample with the present isolated polypeptide, fragment thereof, and/or epitope thereof. In some embodiments theStrongylus vulgaris infection can be detected during a prepatent period. In further embodiments the subject is a horse.

Protective Efficacy Of Centralized And Polyvalent Envelope Immunogens In An Attenuated Equine Lentivirus Vaccine, Jodi K. Craigo, Corin Ezzelarab, Sheila J. Cook, Chong Liu, David Horohov, Charles J. Issel, Ronald C. Montelaro

Maxwell H. Gluck Equine Research Center Faculty Publications

Lentiviral Envelope (Env) antigenic variation and related immune evasion present major hurdles to effective vaccine development. Centralized Env immunogens that minimize the genetic distance between vaccine proteins and circulating viral isolates are an area of increasing study in HIV vaccinology. To date, the efficacy of centralized immunogens has not been evaluated in the context of an animal model that could provide both immunogenicity and protective efficacy data. We previously reported on a live-attenuated (attenuated) equine infectious anemia (EIAV) virus vaccine, which provides 100% protection from disease after virulent, homologous, virus challenge. Further, protective efficacy demonstrated a significant, inverse, linear relationship …

Sudden Death, Aortic Rupture In Horses, Literature Review, Case Studies Reported And Risk Factors, Abelardo Morales Briceño, Aniceto Mendez, Kimberly Brewer, Charlie Hughes, Thomas Tobin

Maxwell H. Gluck Equine Research Center Faculty Publications

Sudden deaths of horses in multiple equestrian disciplines have been attributed to acute and chronic respiratory and cardiovascular diseases. The aim of this study was to perform a review of aortic rupture in horses analyzing, case studies and assessing risk factors. The literature has reported a total of 137 cases of aortic rupture in horses for 28 years (1986-2014), with approximately five horses dying of aortic rupture per year. Histopathologically, there are observed discrete macroscopic degenerative changes in the intima layer only in the aorta. The histological evaluation in the beginning portion of the aorta of the heart evidenced degenerative …

Glucocorticoid-Induced Chondrocyte Cytotoxicity At Doses Recommended For Intra-Articular Therapy In Horses, Wenying Zhu

Theses and Dissertations--Veterinary Science

Intra-articular glucocorticoid injections are commonly used to treat synovitis and osteoarthritis in horses. These agents are highly effective at relieving pain, swelling, and other symptoms of joint inflammation. The drugs also have therapeutic benefits by down regulating the expression of cytokines and protease enzymes that participate in the degradation of articular cartilage. However, detrimental effects on chondrocyte function and cell viability that is independent of osteoarthritis pathogenesis have been described and linked to glucocorticoid use. These side effects are both drug- and dose-dependent. This study tested the hypothesis that manufacture recommended dosage levels of methylprednisolone, betamethasone, and triamcinolone that are …

A Toxicology And Clinical Study Of Post Race Epitaxis Associated With Exercise Induced Pulmonary Hemorrhage In Thoroughbred Race Horses At The Racecourse Rinconada, Caracas, Venezuela, Abelardo Morales Briceno, Diana Villoria Leon, Kimberly Brewer, Thomas Tobin

Maxwell H. Gluck Equine Research Center Faculty Publications

The aim of this study was to describe cases of epistaxis associated with post-race Exercise Induced Pulmonary Hemorrhage (EIPH) in Thoroughbred horses at the Hippodrome "La Rinconada", Caracas, Venezuela, through a clinic pathological study. We studied a total of 29 cases of epistaxis post-race in Thoroughbred horses at the Hippodrome La Rinconada, Caracas, Venezuela, which is 2,950 meters above sea level. The study included horses between the ages of 2-5 years, 16 stallions and 13 mares, weighing between 450-510 kg. They underwent a clinical examination, although horses presenting with epistaxis were in an emergency situation. Samples of blood and urine …

Genetically Engineered Equine Influenza Virus And Uses Thereof, Peter Palese, Adolfo Garcia-Sastre, Thomas Chambers

Veterinary Science Faculty Patents

The present invention relates, in general, to attenuated equine influenza viruses having an impaired ability to antagonize the cellular interferon (IFN) response, and the use of such attenuated viruses in vaccine and pharmaceutical formulations. In particular, the invention relates to attenuated equine influenza viruses having modifications to an equine NS1 gene that diminish or eliminate the ability of the NS1 gene product to antagonize the cellular IFN response. These viruses replicate in vivo, but demonstrate decreased replication, virulence and increased attenuation, and therefore are well suited for use in live virus vaccines, and pharmaceutical formulations.

Breeding Induced Endometritis In The Mare: The Local Innate Immune Response, Elizabeth Moran Woodward

Theses and Dissertations--Veterinary Science

Uterine inflammation after breeding is considered necessary for the clearance of excess semen and debris from the uterus. A subpopulation of mares fails to clear the inflammation in a timely fashion, and develops a persistent breeding induced endometritis (PBIE). Experiments were preformed to evaluate correlations of PBIE to endometrial quality and age. Mares of advanced age and poor endometrial quality had a higher incidence of PBIE. In addition, mares fluctuated in susceptibility to PBIE from one season to the next. The uterine inflammatory gene expression in susceptible and resistant mares within the first 24 hours after breeding was investigated. The …

Infectious Cdna Clone Of The Modified Live Virus Vaccine Strain Of Equine Arteritis Virus, Udeni B. R. Balasuriya, Peter J. Timoney, Jianquan Wang

Veterinary Science Faculty Patents

An isolated polynucleotide molecule includes a DNA sequence encoding an infectious RNA molecule encoding a modified live viral strain of an Equine arteritis virus, wherein the DNA sequence is SEQ ID NO:1 or a degenerate variant thereof. Also provided are transformed or transfected host cells including that sequence, vectors including the sequence, and isolated infectious RNA molecules encoded by the sequence. Further, a modified DNA sequence encoding an infectious RNA molecule encoding a modified live viral strain of an Equine arteritis virus is provided wherein the DNA sequence is SEQ ID NO:2 or a degenerate variant thereof, including a silent …

An Eiav Field Isolate Reveals Much Higher Levels Of Subtype Variability Than Currently Reported For The Equine Lentivirus Family, Jodi K. Craigo, Shannon Barnes, Baoshan Zhang, Sheila J. Cook, Laryssa Howe, Charles J. Issel, Ronald C. Montelaro

Veterinary Science Faculty Publications

BACKGROUND: Equine infectious anemia virus (EIAV), a lentivirus that infects horses, has been utilized as an animal model for the study of HIV. Furthermore, the disease associated with the equine lentivirus poses a significant challenge to veterinary medicine around the world. As with all lentiviruses, EIAV has been shown to have a high propensity for genomic sequence and antigenic variation, especially in its envelope (Env) proteins. Recent studies have demonstrated Env variation to be a major determinant of vaccine efficacy, emphasizing the importance of defining natural variation among field isolates of EIAV. To date, however, published EIAV sequences have been …

Sarcocystis Neurona Diagnostic Primer And Its Use In Methods Of Equine Protozoal Myeloencephalitis Diagnosis, Clara K. Fenger, David E. Granstrom, Alvin A. Gajadhar, Jitender P. Dubey

Veterinary Science Faculty Patents

An amplification primer and probe which can be used in an in vitro diagnostic test for the presence of S. neurona in equine blood or cerebrospinal fluid. Sarcocystis neurona is responsible for the equine condition of protozoal myelitis. The amplification primer is seventeen nucleotides in length and complementary to a unique section of the small ribosomal subunit of Sarcocystis neurona. The primer encompasses nucleotide positions 1470-1487 of the small ribosomal subunit of S. neurona. The primer has the sequence 5' CCATTCCGGACGCGGGT SEQ ID NO:1.

Formulations And Methods To Treat And Prevent Equine Protozoal Myeloencephalitis, David E. Granstrom, Thomas Tobin

Veterinary Science Faculty Patents

The present invention provides formulations useful to treat EPM. It also provides methods to treat EPM in a horse in need of such treatment, comprising administering a pharmaceutically effective amount of a triazine-based anti-coccidial. Preferred are methods to treat EPM using clazuril, diclazuril, toltrazuril or letrazuril. The present invention also provides methods to prevent S. neurona infection in horses comprising administering a prophylactic amount of a triazine-based anticoccidial. Preferred is a method to prevent S. neurona infection by using clazuril, diclazuril, toltrazuril or letrazuril, alone or in combination with other known therapeutics.

Treatment For Contagious Equine Metritis, Thomas W. Swerczek

Veterinary Science Faculty Patents

A composition is provided for the treatment of contagious equine metritis, a contagious veneral disease of horses, which composition comprises an aqueous solution of dextrose, a buffering mixture to provide a pH of about 3.0 to 4.5, and a carrier, the composition being applied topically to external genitalia of horses.