Single Horse Research Articles

Safety assessment of equine allogeneic tenogenic primed mesenchymal stem cells in horses with naturally occurring tendon and ligament injuries.

Mesenchymal stem cells provide a valuable treatment option in orthopedic injuries in horses. The aim of this study was to evaluate the hematological, biochemical, immunological and immunomodulatory parameters following intralesional treatment with tenogenic primed equine allogeneic peripheral blood-derived mesenchymal stem cells (tpMSCs) in client-owned horses with naturally occurring superficial digital flexor tendon (SDFT) and suspensory ligament (SL) injuries. The immunogenicity and immunomodulatory capacities of tpMSCs were assessed in a modified mixed lymphocyte reaction, including peripheral blood mononuclear cells (PBMCs) of 14 horses with SDFT and SL injuries after treatment with tpMSCs. In a second study, 18 horses with SDFT and SL injuries received either an intralesional injection with tpMSCs (n = 9) or no treatment (n = 9). The tpMSCs did not provoke a cellular immune response (p < 0.001) and were able to immunomodulate stimulated T lymphocytes (p < 0.001) in vitro. Therapeutic use of tpMSCs did not result in relevant hematologic or biochemical abnormalities. Both studies had a small sample size. No statistical analyses were performed in the second study. Fibrinogen was only analyzed in a single horse prior to treatment. Co-incubation of tpMSCs and PBMCs of horses that have been previously exposed to tpMSCs did not elicit a cellular immune response and tpMSCs were able to immunomodulate stimulated T lymphocytes. Intralesional treatment with tpMSCs did not provoke abnormal changes in hematological and biochemical parameters.

Prejudicial findings regarding suitability for intended purpose during pre-purchase examinations in a mixed horse population-A retrospective observational study in the United Kingdom.

Pre-purchase examinations (PPEs) are performed for prospective purchasers of horses to identify prejudicial findings that could make animals unsuitable for an intended use. Although this examination process is often standardised, PPEs remain, in large parts, a subjective procedure. In the United Kingdom, PPEs generally consist of either a two stage (two stage vetting [2SV], i.e., general physical examination at rest and basic trot in-hand) or a five stage-examination (five stage vetting [5SV], i.e., general physical exam at rest and after exercise, lameness evaluation including strenuous exercise with re-evaluation after a period of recovery). To identify the proportion of PPEs with prejudicial findings in a mixed horse population in the United Kingdom and to determine those findings. Retrospective observational study. PPE certificates from three first opinion equine practices were reviewed. Data collected included practice identity, examination format (i.e., 2SV or 5SV), PPE outcome, radiographs obtained (yes/no), purchase price, animal signalment, intended use and prejudicial PPE findings, if indicated. Prejudicial findings were grouped in 10 subcategories. Descriptive statistics were obtained for all parameters. Non-normally distributed numeric data between groups were compared using Wilcoxon rank-sum test. Categorical data were compared between groups using Pearson's chi-squared test or Fisher's exact test. Overall, 133 pre-purchase examination certificates were analysed. Seventy-six horses (57.1%) had prejudicial findings: Sixty-one (68.5%) horses undergoing 5SV and 15 (34.1%) of horses undergoing 2SV. Most horses had lameness as the primary prejudicial finding. Horses with a higher purchase price were more likely to undergo 5SV, undergo pre-purchase radiography, and were more likely to have prejudicial findings identified. Data were obtained retrospectively from PPE certificates from a single mixed horse population and different results may have been obtained by analysing a larger number of animals or PPEs of horses intended for different uses, from different geographical regions or undergoing a different PPE format. The level of competition in the intended discipline for horses was not recorded. Analysed data were limited to the information recorded on the PPE certificates, and the original radiographs, ultrasonography images and other additional diagnostic data were not reviewed. Lameness was the most common prejudicial PPE finding. More expensive horses were more likely to undergo a 5SV, have pre-purchase radiographs obtained and have prejudicial findings identified. These results may help inform prospective studies examining the merits of 5SV versus 2SV formats and the value of inclusion of additional diagnostic imaging in PPEs in the general horse population.

FEATURES OF CONSERVATION OF ABANDONED BUILDINGS AMONG THE TRIBES OF THE MAYKOP-NOVOSVOBODNENSKAYA COMMUNITY (ON THE MATERIALS OF NOVOTITAROVSKOYE-14 AND TUZLA-15 SETTLEMENTS)

The article aims to introduce the material on the new Maykop settlement Novotitarovskoye-14 and, to provide the description according to archaeological data of the conservation of the Maykop semi-dugout pits, as well as to discuss the construction of a portable hearth at the settlement Novotitarovskoye-14. The materials are compared with the finds at the Maykop settlement Tuzla-15. The methodology involves the description of the source, the analysis of finds and analogies, the definition of cultural affiliation and chronology. The article also provides the results of radiocarbon dating of materials from the Tuzla-15 settlement and osteological analysis of materials from Kurgan 12 of the Novotitarovskoye-14 group and the Tuzla-15 settlement. The Novotitarovskoye-14 settlement was discovered in 2014 by N.F. Shevchenko, the Tuzla-15 settlement was excavated by S.N. Korenevsky in 2012-2015 and described in a monograph. On both settlements, semi-dugout pits filled with pottery shards, animal bones and small tools in the form of pebbles, fragments of grain grinders were uncovered. At the settlement of Novotitarovskoye-14, the adobe furnace hearth, as well as a large fragment of the brazier were recorded. These findings prove that the Maykop population might have used portable hearths, which were lit outside their buildings. In both settlements, in the filling of abandoned buildings, a layer of ash from a fire that burned outside was recorded. The analysis of osteological findings clearly reflects that the majority of bones belong to large and small cattle. Single horse bones were found at Novotitarovskoye-14 settlement. The settlement of Novotitaroovskoye-14 belongs to the Maykop variant of the Maykop-Novosvobodnenskaya community, and the settlement of Tuzla-15 belongs to the similar Psekup variant of the Maykop-Novosvobodnenskaya community. The date of the first settlement cannot be determined precisely within the framework of the existence of the Maykop sites. The date of settlement of Tuzla-15 belongs to the final stage of the Maykop-Novosvobodnenskaya community.

FEATURES OF CONSERVATION OF ABANDONED BUILDINGS AMONG THE TRIBES OF THE MAYKOP-NOVOSVOBODNENSKAYA COMMUNITY (ON THE MATERIALS OF NOVOTITAROVSKOYE-14 AND TUZLA-15 SETTLEMENTS)

The article aims to introduce the material on the new Maykop settlement Novotitarovskoye-14 and, to provide the description according to archaeological data of the conservation of the Maykop semi-dugout pits, as well as to discuss the construction of a portable hearth at the settlement Novotitarovskoye-14. The materials are compared with the finds at the Maykop settlement Tuzla-15. The methodology involves the description of the source, the analysis of finds and analogies, the definition of cultural affiliation and chronology. The article also provides the results of radiocarbon dating of materials from the Tuzla-15 settlement and osteological analysis of materials from Kurgan 12 of the Novotitarovskoye-14 group and the Tuzla-15 settlement. The Novotitarovskoye-14 settlement was discovered in 2014 by N.F. Shevchenko, the Tuzla-15 settlement was excavated by S.N. Korenevsky in 2012-2015 and described in a monograph. On both settlements, semi-dugout pits filled with pottery shards, animal bones and small tools in the form of pebbles, fragments of grain grinders were uncovered. At the settlement of Novotitarovskoye-14, the adobe furnace hearth, as well as a large fragment of the brazier were recorded. These findings prove that the Maykop population might have used portable hearths, which were lit outside their buildings. In both settlements, in the filling of abandoned buildings, a layer of ash from a fire that burned outside was recorded. The analysis of osteological findings clearly reflects that the majority of bones belong to large and small cattle. Single horse bones were found at Novotitarovskoye-14 settlement. The settlement of Novotitaroovskoye-14 belongs to the Maykop variant of the Maykop-Novosvobodnenskaya community, and the settlement of Tuzla-15 belongs to the similar Psekup variant of the Maykop-Novosvobodnenskaya community. The date of the first settlement cannot be determined precisely within the framework of the existence of the Maykop sites. The date of settlement of Tuzla-15 belongs to the final stage of the Maykop-Novosvobodnenskaya community.

Recovery of Salmonella bacterial isolates from pooled fecal samples from horses.

It is important to determine if a horse is shedding Salmonella spp., but a complete culture series can be cost prohibitive. Determine the optimal pooling technique to maintain high sensitivity of Salmonella spp. culture using spiked samples, and then demonstrate the efficacy of this protocol on clinical submissions. Pooled fecal samples are as sensitive as 5 individual cultures for the detection of Salmonella shedding. A single Salmonella-negative horse from the university herd, and 19 hospitalized horses. Salmonella-free fecal samples were spiked with different amounts of Salmonella spp. (102 , 103 , 104 , and 105 colony forming units [cfu]) and homogenized to evaluate pooled samples. Five individual fecal samples were collected from 19 hospitalized horses. Ten-gram aliquots of each individual sample were combined to make a pooled sample. Both individual and pooled samples were cultured for Salmonella spp. The identity of bacterial isolates was confirmed by matrix-assisted laser desorption-ionization time of flight mass spectrometry. A 102 cfu concentration of Salmonella spp. could be recovered from a spiked Salmonella-free fecal sample. Homogenization protocols indicated that the addition of 20 mL of broth to the pooled sample improved recovery, whereas homogenization time did not. Of the 19 horses tested, 5 were positive for Salmonella. In all instances, Salmonella spp. were recovered from the fecal pool as well as individual samples. Pooling of 5 fecal samples for Salmonella culture is a sensitive and cost-effective diagnostic approach to detect horses that are shedding the organism.

Nasal transmission of equine parvovirus hepatitis.

Equine parvovirus hepatitis (EqPV-H) is highly prevalent and causes subclinical to fatal hepatitis, which can occur in outbreaks. Whereas iatrogenic transmission is well documented, the mode of horizontal transmission is not known. The virus is shed in nasal, oral and fecal secretions, and PO transmission has been reported in a single horse. Investigate the efficiency of PO and nasal transmission of EqPV-H in a larger cohort. Prospective experimental transmission study. Eleven EqPV-H-negative horses were inoculated with 5 × 106 genome equivalents EqPV-H. Serum PCR and serology for EqPV-H were performed weekly and monthly, respectively. Horses first were inoculated PO, and then intranasally 8 weeks later. No horse became viremic or seroconverted within 8 weeks after PO inoculation. After intranasal inoculation, 5 horses became viremic within 6 to 12 weeks and seroconverted within 10 to 19 weeks. After a period without monitoring from 12 to 19 weeks postinoculation, another 5 horses were found to be viremic at 19 to 22 weeks. The second set of 5 horses could have been infected by horizontal transmission from the first 5 because of cohousing. We demonstrated that EqPV-H can be transmitted nasally. The prolonged eclipse phase before detectable viremia indicates biosecurity measures to control spread could be impractical.

Riding to the rescue: an addition to the Plains Biographic rock art lexicon

Depictions of two riders mounted on a single horse appear in Plains Biographic rock art at five sites. Comparison of these images with late nineteenth century Plains Indian drawings on hide and paper indicates that the pictographs and petroglyphs commemorate instances in which mounted warriors rescued comrades who had been unhorsed in battle. Ethnographic material confirms that several Plains Indian societies held this deed in high esteem. These societies employed formal mechanisms for recognizing individuals who had rescued comrades. While the importance of martial themes in Plains Biographic rock art has been widely acknowledged, emphasis has been placed on the acts of counting coup, capturing weapons, and taking horses. The identification of depictions of two warriors on a single horse as rescue scenes adds to our understanding of the diversity of martial exploits recognized by Plains Indian societies.

Characterisation of faecal microbiota in horses medicated with oral doxycycline hyclate.

Antimicrobial-associated diarrhoea is a common adverse effect of antimicrobial treatment in horses and has been reported following the administration of oral doxycycline. The administration of antimicrobials has also been associated with changes in the equine intestinal microbiota diversity yet has not been explored under doxycycline treatment. To describe the dynamics of the faecal microbial diversity following a 5-day oral administration of doxycycline in healthy horses with Streptococcus zooepidemicus infected tissue chambers. Experimental prospective cohort study in a single horse group. Seven healthy adult horses with S. zooepidemicus infected tissue chambers received oral doxycycline at 10mg/kg q 12h for 5-days following the tissue chamber inoculation. Faeces were collected prior to the tissue chamber inoculation and until 28-days post inoculation. Faecal microbiota was characterised by high throughput sequencing of the V4 region of the 16S rRNA gene on the Illumina MiSeq sequencing platform. Bioinformatic analysis was performed with Mothur and statistical analysis were conducted on R Studio. A significant decrease in alpha diversity, characterised by a decrease of richness and diversity, and a decrease in beta diversity, characterised by changes in relative abundance, occurred after initiation of and during the administration of doxycycline. A decrease in Verrucomicrobia and increase in Firmicutes:Bacteroidetes ratio occurred following the initiation of treatment, with a return to initial Firmicutes:Bacteroidetes ratio during the treatment. It took 23days after discontinuing the treatment for the faecal microbiota to return close to the initial state. Lack of control population within the study. Transitory intestinal dysbiosis occurs under oral administration of doxycycline in horses.

The potential and limits of Thin-Plate Spline retrodeformation on asymmetrical objects:simulation of taphonomic deformations andapplication on a fossil sample of limb long bones

In this study, we suggest a method adapted to the retrodeformation of asymmetrical objects – such as limb bones – by quantitatively estimating the effectiveness of the Thin-Plate Splines (TPS) interpolation function as a retrodeformation tool. To do so, taphonomic deformations were first simulated on a single horse femur. The original bone was then used as a reference in order to drive the retrodeformation using anatomical landmarks. This approach, based on a single bone, enabled us to evaluate the performance of the retrodeformation procedure. Then, the same approach was performed on a sample of rhino femora but using a different specimen (from the same species) as the reference in order to account for morphological variation. We also added sliding semi-landmarks on anatomical curves. Finally, retrodeformation was applied on a sample of sauropodomorph dinosaur femora by building a mean shape based on several well-preserved fossil specimens. Results show that entirely flattened and stretched bones are more efficiently retrodeformed than bent and twisted bones. Introduction of morphological variation increased the efficiency of retrodeformation for bent and locally stretched bones. The application to the sample of fossils produced similar results but also highlighted the difficulty of retrodeforming bones with a combination of different deformations. TPS interpolation is an efficient tool of retrodeformation for asymmetrical objects, especially for bones with only one affine deformation such as flattening or stretching. Finding a threshold of landmark number to use for this process would be the next step because it would allow us to ensure the quality of retrodeformation while keeping available a reasonable number of landmarks in order to perform shape analysis on retrodeformed bones. Twisted and bent fossils are frequently discovered and we suggest that these kinds of deformations should be studied with caution, especially when combined with other types of taphonomic distortions.

Investigation of the Role of Healthy and Sick Equids in the COVID-19 Pandemic through Serological and Molecular Testing.

Simple SummaryThe objective of the present study was to determine if horses are susceptible to SARS-CoV-2. Nasal swabs from 667 equids with acute onset of fever and respiratory signs were tested by qPCR for SARS-CoV-2. Further, 633 serum samples collected from a cohort of 587 healthy racing Thoroughbreds with possible exposure to humans with SARS-CoV-2 infection were tested for antibodies to SARS-CoV-2 using an ELISA targeting the receptor-binding domain of the spike protein. All 667 horses with fever and respiratory signs tested qPCR-negative for SARS-CoV-2. A total of 35/587 (5.9%) Thoroughbred racing horses had detectable IgG antibodies to SARS-CoV-2. While horses appear to be susceptible to SARS-CoV-2 when in close contact with humans with SARS-CoV-2 infection, clinical disease was not observed in the study horses. Experimental challenge studies using pure inocula are needed in order to study the clinical, hematological, molecular, and serological features of adult horses infected with SARS-CoV-2.More and more studies are reporting on the natural transmission of SARS-CoV-2 between humans with COVID-19 and their companion animals (dogs and cats). While horses are apparently susceptible to SARS-CoV-2 infection based on the homology between the human and the equine ACE-2 receptor, no clinical or subclinical infection has yet been reported in the equine species. To investigate the possible clinical role of SARS-CoV-2 in equids, nasal secretions from 667 horses with acute onset of fever and respiratory signs were tested for the presence of SARS-CoV-2 by qPCR. The samples were collected from January to December of 2020 and submitted to a commercial molecular diagnostic laboratory for the detection of common respiratory pathogens (equine influenza virus, equine herpesvirus-1/-4, equine rhinitis A and B virus, Streptococcus equi subspecies equi). An additional 633 serum samples were tested for antibodies to SARS-CoV-2 using an ELISA targeting the receptor-binding domain of the spike protein. The serum samples were collected from a cohort of 587 healthy racing Thoroughbreds in California after track personnel tested qPCR-positive for SARS-CoV-2. While 241/667 (36%) equids with fever and respiratory signs tested qPCR-positive for at least one of the common respiratory pathogens, not a single horse tested qPCR-positive for SARS-CoV-2. Amongst the racing Thoroughbreds, 35/587 (5.9%) horses had detectable antibodies to SARS-CoV-2. Similar to dogs and cats, horses do not seem to develop clinical SARS-CoV-2 infection. However, horses can act as incidental hosts and experience silent infection following spillover from humans with COVID-19. SARS-CoV-2-infected humans should avoid close contact with equids during the time of their illness.

Morphological Characteristics of a Horse Discovered in an Avar-Period Grave from Sâncraiu de Mureș (Alba County, Romania).

Simple SummaryThe present study deals with the morphological description of the animal remains in an Avar grave that contained the remnants of a horse burial, along with its owner, in a cemetery situated in Sâncraiu de Mures, Alba County, Romania, located geographically in the Transylvanian Plateau. The investigation of the grave and the data about the remains were published by archaeologists from the Institute of Archaeology of Cluj Napoca, Romania. For analysis and study, the animal bone remains were sent to the Faculty of Veterinary Medicine Cluj-Napoca. After a careful analysis of the morphological characteristics of the bones, the conclusions showed that the bones belonged to one young horse of 2–2.5 years, with an average recalculated shoulder height as being situated in the 1300–1400 mm interval. The study provides a set of morphological data regarding the characteristics of the horses from this scantily represented history in Romania, and the results show that the data obtained are consistent with the similar Avar horse burials in the region.Archeozoological studies provide an insight into human–environment relations, bringing important information on the morphology of the animal and the role of the animal and its functions. The purpose of this study was to reveal the morphological characteristics of the horse identified in an 8th century BC Avar cemetery dated, as it resulted from the investigation carried out on the materials presented to the Anatomy Lab of Department of Comparative Anatomy. The cleaning and conditioning of the materials were performed in the lab, followed by anatomical and osteometrical study. The identification of the species, the osteometrical interpretation and the assessment of age at death constituted the basis of the main conclusions. Based on the morphological and metrical data, we concluded that the fragments originated from a single young horse individual (Equus caballus) no older than 2.5 (2–2.5 years age span) included into class 5 of height (Vitt scale) with slender extremities. An overview of the available data (horse morphology) from similar sites in Romania and neighboring territories (Hungary and Croatia) is presented, with the intention of a general framing for the characteristics of the horses used by the Avar populations.

LC-HRMS/MS study of the prodrug ciclesonide and its active metabolite desisobutyryl-ciclesonide in plasma after an inhalative administration to horses for doping control purposes.

Ciclesonide (CIC) is the first inhaled highly potent corticosteroid that does not cause any cortisol suppression. It has been developed for the treatment of asthma in human and more recently in equine. CIC is the active compound of Aservo® EquiHaler® (Boehringer Ingelheim Vetmedica GmbH), the pre-filled inhaler generating a medicated mist based on Soft Mist™ technology. This prodrug is rapidly converted to desisobutyryl-ciclesonide (des-CIC), the main pharmacologically active compound. Due to its anti-inflammatory properties, CIC is prohibited for use in horse competitions. To set up an appropriate control, the determination of detection times and screening limits are required. Therefore, a highly sensitive analytical method based on supported liquid extraction (SLE) combined with liquid chromatography-high resolution tandem mass spectrometry (LC-HRMS/MS) was developed to detect CIC and its active metabolite des-CIC in plasma. The lower limit of detection of CIC and des-CIC was approximately 1 pg/ml in plasma. After a pilot study conducted on a single horse at the recommended dose (eight actuations twice daily corresponding to 5.5mg/day for the first 5days, followed by 12 actuations once daily corresponding to 4.1mg/day in the last 5days), the same protocol was applied in the main study using six horses. In all horses, CIC and des-CIC levels were less than 5 and 10pg/ml, respectively, at 36 h after the end of the administration. The outcome of this risk assessment study should be useful to draw any recommendations for horse competitions.

Mission Critical: Caremark, Blue Bell, and Director Responsibility for Cybersecurity Governance

If the potential for Caremark liability hangs like the sword of Damocles over corporate directors of Delaware corporations, then that sword has been considerably more secure than that of the original myth. For decades, Chancellor Allen’s description of a Caremark claim as “possibly the most difficult theory in corporation law upon which a plaintiff might hope to win a judgment” held true. Caremark claims that survived a motion to dismiss were for decades few and far between. That changed in 2019. In the space of little over a year, Delaware courts have allowed five Caremark claims to survive in the corporation context and one claim to survive in the limited partnership context. The thread holding that sword is beginning to look more like the single horse hair of myth. The scope and likelihood of Caremark liability are matters of considerable interest and concern for directors. Under most circumstances, a board simply doing its job poorly is relevant only to the directors’ duty of care and protected by the business judgment rule, exculpatory provisions under Section 102(b)(7), and advancement and indemnification. Failure to monitor under Caremark, however, is a breach of the duty of loyalty. A breach of the duty of loyalty is not protected by the business judgment rule. It cannot be exculpated. And it cannot be covered by indemnification. 2019 marked an abrupt shift in Caremark in application, if not in theory. In June of that year the Supreme Court of Delaware (“Delaware Supreme Court”) reversed a decision by the Delaware Court of Chancery (“Chancery Court”) dismissing a claim against the directors of Blue Bell Creameries, Inc. (“Blue Bell”) under Caremark. Within a little over a year, the Chancery Court would sustain Caremark claims in four cases. In Clovis, the Chancery Court sustained a Caremark claim against directors of a pharmaceutical company who allowed the company to misrepresent the clinical trial success of one of its three drugs. In Hughes, the Chancery Court sustained a Caremark claim against directors of a Chinese company incorporated in Delaware that suffered from severe and pervasive accounting issues. In Teamsters Local, the Chancery Court sustained a Caremark claim against directors of a large pharmaceutical company who allowed an indirect subsidiary to essentially operate a criminal enterprise. And in Boeing, the Chancery Court sustained a Caremark claim against directors of an airplane manufacturer who did not pay attention to safety issues. Cybersecurity has become just such a mission critical risk for large corporations. This Article makes four key arguments. Black letter Caremark doctrine has not changed, but it is newly reinvigorated and the risks of Caremark liability for directors is greater than just a few years ago. Future Caremark liability will be centered on failure to provide board-level oversight of mission critical risks. Cybersecurity is mission critical to effectively all large companies today. The risk of Caremark liability can be mitigated by taking a few simple steps to ensure that the board is addressing cybersecurity. This Article is the first to make these arguments together and the first to make the final argument.

Phosphorus Non-Point Pollution from Equestrian Wastes and the Need for Recycling

South Florida and much of the rest of the World suffers from harmful algal blooms (HABs) and controls of both nitrogen (N) and phosphorus (P) pollution are required to curtail the onset, spread and/or expansion of these blooms.&#x0D; &#x0D; This report covers our studies on several aspects of equestrian waste (viz. horse manure) aimed at yielding an overview of phosphorus and its pollution stemming from non-point horse manure sources in portions of Palm Beach County Florida. Methods included a modified Hedley extraction sequence, emphasizing &amp;lsquo;easily extractable phosphorus&amp;rsquo; (EEP), and 31P nuclear magnetic resonance (NMR) spectroscopic identification of organic phosphorus (Po) species.&#x0D; &#x0D; Samples included fresh and aged horse manure, pasture soils, horse feed and pasture grasses, and canal waters adjacent to equestrian or agricultural fields.&#x0D; &#x0D; Easily extractable Phosphorus (EEP) averaged about 54-77% of the total horse manure phosphorus. Total phosphorus ranged from 13,020 &amp;ndash; 22,300 mg per kilogram dry weight. (&amp;asymp;60-100 lbs. P2O5 / ton and on a wet weight basis, this equates to 4,000 to 14,818 grams-P/ U.S. ton or 8.8 to 32.6 pounds of phosphorus (&amp;asymp; 20-75 lb. P2O5) per wet weight ton of horse manure. Considering the values of EEP in fresh samples from a single horse, we found a range of 8,000 &amp;ndash; 17,000 mg-P/kg (8-17 g-P/kg) dry weight horse manure.&#x0D; &#x0D; Soil samples yielded the highest P in the NaOH extract of the Hedley sequence. This equates to the Al, Fe and ester forms. &#x0D; &#x0D; Phosphorus (viz. EEP) runoff is viewed here as a non-point P pollution source.

18  Relative expression of the neonatal crystallizable fragment receptor (FcRn) in equine tissues

The neonatal crystallizable fragment receptor (FcRn) is an important immune system receptor with several functions including bi-directional transport of immunoglobulin G across cell surfaces and involvement in antigen presentation. Research regarding the FcRn is robust in human medicine, but has lagged in livestock species. Although the FcRn has been characterized in several body tissues in cattle, sheep and pigs, there is only one published report regarding examination of the FcRn in intestine from a single horse in China. Therefore, the objective of this study was to identify relative FcRn mRNA expression in various equine body tissues. To meet the objective, tissues were collected from 2 horses (H1, H2) within 30 min of euthanasia. Collection sites were standardized for each location to minimize variation. Tissues included lung (4 locations), stomach (2 locations), small intestine (3 locations), cecum (3 locations), large colon (6 locations), and joints (2 locations). A minimum of 5 tissue samples (500 mg each) were collected from each location to obtain representative samples and preserved in RNAlater. Total RNA was extracted using the RNeasy Plus Universal Mini Kit (Qiagen). Reverse transcription to cDNA was performed with the High-Capacity cDNA Reverse Transcription kit (ThermoFisher Scientific). Pre-designed TaqMan probe sets for FcRn (Assay ID 07049868_g1) and 3 reference genes (ACTB, GADPH, HPRT1) were obtained from ThermoFisher Scientific. Relative quantification of mRNA expression of FcRn in each tissue was measured by qPCR using TaqMan Fast Advanced Master Mix (ThermoFisher Scientific) and calculated using the 2 −ΔΔCT method and normalized to the geometric mean of the 3 reference genes. Realtive quantity (RQ) of FcRn expression for tissues from the 2 horses was grouped qualitatively into high, moderate, and low designations. RQ of FcRn expression was highest in the joints (RQ range 2.37–3.98) and small intestine (RQ range 2.28–3.18) with individual variation present in the duodenum (H1 RQ 2.60; H2 RQ 0.82) and ileum (H1 RQ 1.08; H2 RQ 2.47). FcRn expression was moderate in tissues from the cecum (RQ range 1.02–1.75) and lung (RQ 1.42–2.32) with individual variation present in the distal cecum (H1 RQ 1.02; H2 RQ 0.67) and main bronchi tissue (H1 RQ 2.06; H2 RQ 0.47). Expression was lowest in the stomach (RQ range 0.24–1.12) and colon (RQ range 0.56–1.35). FcRn location within specific cell types was not determined and requires further examination. In summary, this novel qualitative study indicates FcRn expression in numerous equine tissues and provides base knowledge from which to expand future investigation.

30  An investigation into the relationship between rider pelvic asymmetry and equine pelvic asymmetry in relation to the use of physical therapy

There is increasing use of physical therapy for both horse and rider but limited scientific research linking the horse, the rider and their symmetry together. Pelvic asymmetry can indicate musculoskeletal imbalance and affect performance. Therapy may have an impact on the pelvic asymmetry of both the horse and the rider as independent parties. This study aimed to investigate the hypothesis that single horse/rider combinations who both have regular (4 or more times per year) physical therapy will have improved pelvic symmetry rather than horse/rider combinations that have no physical therapy. Fifty single horse (age 6–14yrs) and rider (age 18–45yrs) combinations (minimum 6-mo partners) were selected with inclusion/exclusion criteria. Treatment group (n = 25) horse and rider received regular physical therapy (chiropractic, osteopathy, physiotherapy or sports massage therapy). Control group (n = 25) pairs received no therapy ever or over 1 year prior. Data collection was blinded to groups. On level ground, triplicate measurements of horse tuber coxae (TC) heights (stood square) and rider anterior superior iliac spine (ASIS) (feet@30cm) to floor were taken using a plumb line. Symmetry indices (SI) were calculated from raw data. Data were tested for normality (Shapiro Wilk test) and Mann Whitney test, regression analysis tests used with level of statistical significance at P < 0.05. There was a significant difference between treatment and control groups of pelvic symmetry values for horses (mean SI ± SD: control 4.69 ± 2.48, treatment 2.35 ± 0.61, P = 0.00001) and riders (mean SI ± SD: control 3.4 ± 1.25, treatment 2.11 ± 0.88, P = 0.00003). There was no significant correlation between horse TC and rider ASIS asymmetry means for treatment group (r 2 = 0.04, F = 0.95, P = 0.34) or control group (r 2 = 0.01, F = 0.29, P = 0.6). This study provides positive evidence that regular use of physical therapy for horses and riders may improve pelvic asymmetry of the horse and rider individually. This is a positive basis for further research into the casual effect of relationships between the horse and rider and their use of physical therapy.

PSV-36 Optimization of Collection, Storage, and Extraction of Samples for Large-Scale 16s Sequencing Project

Abstract Correlations between the microbiome of the horse and various physiological parameters continue to be determined. Most evaluations of the equine microbiome have been regionally focused with limited numbers or variability. In order to elucidate the role of the microbiome on equine health, a large-scale trial has been initiated with a primary objective of analyzing and characterizing the microbiome of the horse. A study of this scope relies on the horse owner to collect samples from various locations and ship them to a centralized lab. Therefore, collection protocols and extraction techniques need to be optimized to maintain sample integrity. A trial was conducted to evaluate the storage conditions and extraction methodologies necessary to develop an optimized method of sample collection, preparation, and extraction. Samples were collected from a single horse and subjected to one of four treatments in duplicate (1-fresh, 2-frozen, 3-stored in 99% EtOH, and 4-stored in a commercially available DNA/RNA transport medium; DNA/RNA Shield®, Zymo Research, Irvine, CA). Frozen samples were placed at -80◦ C immediately following collection and stored for 48-hr. Frozen samples were thawed immediately prior to extraction. All other treatments were stored as dictated by treatment at room temperature for 48-hr prior to extraction to mimic shipping. Samples were processed utilizing a commercially available DNA extraction kit (Quick-DNA™ Fecal/Soil Microbe Miniprep, Zymo Research). DNA extract was analyzed for quantity utilizing a Qubit Fluorometer (ThermoFisher, Waltham, MA), and quality via spectrophotometry (NanoDrop 2000; ThermoFisher, Waltham, MA). Swabs stored in DNA/RNA Shield® yielded higher concentrations of DNA with superior 260/280 values than other treatments. These data indicate that storage of fecal swabs in an appropriate buffer combined with optimized extraction protocols can deliver adequate extracts that can be utilized for 16s sequencing and validates the use of this method for large-scale microbiome analysis.

High genetic diversity of ancient horses from the Ukok Plateau.

A growing number of researchers studying horse domestication come to a conclusion that this process happened in multiple locations and involved multiple wild maternal lines. The most promising approach to address this problem involves mitochondrial haplotype comparison of wild and domestic horses from various locations coupled with studies of possible migration routes of the ancient shepherds. Here, we sequenced complete mitochondrial genomes of six horses from burials of the Ukok plateau (Russia, Altai Mountains) dated from 2.7 to 1.4 thousand years before present and a single late Pleistocene wild horse from the neighboring region (Denisova cave). Sequencing data indicates that the wild horse belongs to an extinct pre-domestication lineage. Integration of the domestic horse data with known Eurasian haplotypes of a similar age revealed two distinct groups: the first one widely distributed in Europe and presumably imported to Altai, and the second one specific for Altai Mountains and surrounding area.

The equine diaphragm: A novel technique for repeatable ultrasound measurement.

The diaphragm is an important respiratory muscle, playing a key role during exercise. In humans, diaphragm thickness increases in response to training and is correlated with inspiratory strength. In order to assess respiratory strength in the horse, new evaluation techniques are required and measurement of diaphragm thickness, in a non-invasive and repeatable manner, is a possible approach. The purposes of this four-part, prospective, pilot study were to develop and describe a repeatable method to measure the diaphragm thickness, using ultrasonography, in Thoroughbred racehorses. A standardized technique was developed whereby the ultrasound transducer was positioned 1cm below a line between the cranioventral aspect of the tuber coxae and olecranon. The diaphragm thickness was measured on three occasions 1week apart, by a single observer to determine the intraobserver repeatability, and by a second observer on one occasion to assess interobserver reproducibility. The diaphragm was observed in all intercostal spaces (ICS) from 7 to 17 on the left side, and 6 to 17 on the right side in a single horse. The thickest measurement (1.42cm), obtained from 11 horses, was at ICS 11 on the left-side during inspiration. The narrowest measurement (0.56cm) was obtained at ICS 16 on the right-side during expiration. There was no significant difference between the measurements obtained by a single observer on three occasions (P<.05). This is the first study to provide a detailed description of ultrasonographic imaging and measurement of the equine diaphragm. The novel technique developed to position the ultrasound transducer in a standardized location allowed examination and measurement of the diaphragm with good repeatability.

Marisol’s Antimonument: Masculinity, Pan-Americanism, and Other Imaginaries

Marisol’s assemblage The Generals (1961–62) assumes a guise of well-worn signifiers of midcentury US patriotic masculinity: equestrian statue, founding father, soldier, and cowboy. At the same time, this sculpture of George Washington and Simón Bolívar on a single horse invokes the very forces Cold Warriors vilified as un-American threats at home and abroad: homoeroticism and Latin American dissent. Marisol’s irreverent antimonument, which has garnered little analysis but performed a central role during her meteoric rise in the 1960s, tapped into Cold War discourses about sexual politics, freedom, national mythologies, and inter-American relations.